EPA/635/R-08/012A

DEVELOPMENT OF A RELATIVE POTENCY 

FACTOR (RPF) APPROACH FOR POLYCYCLIC 

AROMATIC HYDROCARBON (PAH) MIXTURES 

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September 2009 

NOTICE 

This document is an interagency science consultation draft. This information is distributed 

solely for the purpose of pre-dissemination peer review under applicable information quality 

guidelines. It has not been formally disseminated by EPA. It does not represent and should not 

be construed to represent any Agency determination or policy. It is being circulated for review 

of its technical accuracy and science policy implications.

U.S. Environmental Protection Agency 

Washington, DC 
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DISCLAIMER 

This document is a preliminary draft for review purposes only. This information is 

distributed solely for the purpose of pre-dissemination peer review under applicable information 

quality guidelines. It has not been formally disseminated by EPA. It does not represent and 

should not be construed to represent any Agency determination or policy. Mention of trade 

names or commercial products does not constitute endorsement or recommendation for use. 
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EXECUTIVE SUMMARY 

The U.S. Environmental Protection Agency’s (U.S. EPA’s) Integrated Risk Information 

System (IRIS) Program is releasing for scientific review a relative potency factor (RPF) 

approach for polycyclic aromatic hydrocarbon (PAH) mixtures as one approach to assessing 

cancer risk from exposure to PAH mixtures. The RPF approach is not a reassessment of 

individual PAH carcinogenicity, but rather provides a cancer risk estimate for PAH mixtures by 

summing doses of component PAHs after scaling the doses (with RPFs) relative to the potency 

of an index PAH (i.e., benzo[a]pyrene). The cancer risk is then estimated using the doseresponse curve for the index PAH. RPFs for seven individual PAHs were developed in the U.S. 

EPA (1993) Provisional Guidance for Quantitative Risk Assessment of PAHs (Provisional 

Guidance) and are utilized extensively within U.S. EPA program offices and other regulatory 

agencies. The Provisional Guidance, however, does not reflect the most recent research, nor 

does it consider additional PAHs with carcinogenic potential (such as fjord-region PAHs). 

The Supplemental Guidance for Conducting Health Risk Assessment of Chemical 

Mixtures (U.S. EPA, 2000) highlights that approaches based on whole mixtures are preferred to 

component approaches, such as the RPF approach. Risk assessment approaches based on 

toxicity evaluations of whole mixtures inherently address specific interactions among PAHs and 

account for the toxicity of unidentified components of PAH mixtures. They also do not require 

assumptions regarding the toxicity of individual components (e.g., dose additivity or response 

additivity). While whole mixture assessment is preferred, there are challenges associated with 

using these approaches. There are very few toxicity data available for whole PAH mixtures and, 

in most cases, chemical analyses of the composition of mixtures are limited. In addition, PAHcontaining mixtures tend to be very complex; the composition of these mixtures appears to vary 

across sources releasing these mixtures to the environment and in various environmental media 

in which they occur. For these reasons, a whole mixtures approach may not always be 

practicable for risk assessment purposes. 

This report provides recommendations for development of the RPF approach for PAH 

mixtures health risk assessment and includes: 

(1) A rationale for recommending an RPF approach (Section 2); 

(2) A summary of previous approaches for developing the RPF approach for PAHs 

(Section 3); 

(3) An evaluation of the carcinogenic potential of individual PAHs (Section 4); 
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(4) Methods for dose response assessment and individual study RPF calculation (Section 5); 

(5) Selection of PAHs for inclusion in the RPF approach (Section 6); 

(6) Derivation of RPFs for selected PAHs (Section 7); and 

(7) Characterization of strengths, weaknesses, and uncertainties associated with the RPF 

approach to PAH cancer risk assessment (Section 8).

The RPF approach involves two key assumptions: (1) similar toxicological action of 

PAH components in the mixture; and (2) interactions among PAH mixture components do not 

occur at low levels of exposure typically encountered in the environment (that is, additivity is 

assumed). Mechanistic studies indicate that the mutagenic and tumor-initiating activity of 

carcinogenic PAHs requires metabolic activation to reactive intermediates (e.g., dihydrodiol 

epoxides, quinones, radical cations), which covalently modify DNA targets resulting in mutation, 

and that tumor promotion and progression phases may involve parent compound binding to the 

Ah receptor (AhR) and subsequent alterations of gene expression or a cell proliferation response 

to metabolite cytotoxicity (see Section 2.4, Similarities in Carcinogenic Mode of Action for 

PAHs, and Figure 2-3, Overview of the Proposed Key Events in the Mode of Action for PAH 

Carcinogenicity). As such, there is evidence that an assumption of similar toxicological action is 

reasonable; however, the carcinogenic process for PAHs is likely to be related to some unique 

combination of multiple molecular events resulting from formation of several reactive species. 

The second assumption of no interactions at low levels of exposure is reasonable, but evidence of 

toxicological interactions among PAHs at higher dose levels has been observed (see Section 2.7, 

Additivity of PAHs in Combined Exposures). 

Several approaches have been used previously for the determination of RPFs for PAHs 

(see Section 3). In the published literature, RPF values were proposed in at least one analysis for 

a total of 27 PAHs (see Table 3-1). Because these approaches generally relied on similar 

bioassay data and modeling methods, the resulting RPF values are generally comparable for most 

PAHs across studies. The RPF approach provided in the current report makes use of more recent 

data on genotoxicity and tumorigenicity of PAHs. 

There is a large PAH database on carcinogenicity in animal bioassays, genotoxicity in 

various test systems, and bioactivation to tumorigenic and/or genotoxic metabolic intermediates. 

The RPF analysis presented here includes only unsubstituted PAHs with three or more fused 

aromatic rings containing only carbon and hydrogen atoms, because these are the most widely 

studied members of the PAH chemical class. The study types that were considered most useful 

for RPF derivation were rodent carcinogenicity bioassays (all routes) in which one or more 

PAHs was tested at the same time as benzo[a]pyrene. In addition, in vivo and in vitro data for 
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cancer-related endpoints in which one or more PAHs and benzo[a]pyrene were tested 

simultaneously were obtained, including studies on the formation of DNA adducts, mutagenicity, 

chromosomal aberrations, sister chromatid exchange frequency, aneuploidy, DNA 

damage/repair/recombination, unscheduled DNA synthesis, and cell transformation. Although it 

would be possible to calculate RPFs from studies where a PAH and benzo[a]pyrene were tested 

by the same laboratory using the same test system but at different times, this approach was not 

considered because it could introduce differences in the dose-response information that are 

unrelated to the chemical (e.g., variability associated with laboratory environment conditions, 

animal handling, food supply, etc.). Thus, studies in which benzo[a]pyrene was not tested 

simultaneously with another PAH were not considered in the RPF calculations. 

Studies of AhR binding/activation were not considered for use in deriving RPFs because 

there is evidence indicating that highly mutagenic fjord-region PAHs are potent carcinogens 

despite exhibiting lower AhR affinity (reviewed by Bostrom et al., 2002). Likewise, some PAHs 

that strongly activate the AhR, such as benzo[k]fluoranthene (Machala et al., 2001), are only 

weakly carcinogenic. In addition, some studies have demonstrated the formation of DNA 

adducts in the liver of AhR knock-out mice following intraperitoneal or oral exposure to 

benzo[a]pyrene (Sagredo et al., 2006; Uno et al., 2006; Kondraganti et al., 2003), indicating that 

Ah responsiveness is not a prerequisite to genotoxicity. These findings suggest that there may be 

alternative (i.e., non-AhR mediated) mechanisms of benzo[a]pyrene activation in the mouse 

liver, and that AhR affinity would not be a good predictor of carcinogenic potency. 

Several study types were excluded from the database because they did not provide 

carcinogenicity or cancer-related endpoint information for individual PAHs. These include 

biomarker studies measuring DNA adducts in humans, studies of PAH metabolism, and studies 

of PAH mixtures. Although these studies contain important information on human exposure to 

PAH mixtures and the mode of action for PAH toxicity, they generally do not contain doseresponse information that would be useful for calculation of RPF estimates. 

A database of primary literature relevant to the RPF approach for PAHs was developed by 

performing a comprehensive review of the scientific literature dating from the 1950s through 

2009 on the carcinogenicity and genotoxicity of PAHs. The search identified over 900 individual 

publications for a target list of 74 PAHs (see Table 2-1) that have been identified in 

environmental media or for which toxicological data are available. Review of these publications 

resulted in the identification of more than 600 papers that included carcinogenicity or cancerrelated endpoint data on at least one PAH and benzo[a]pyrene tested at the same time. 

References in the PAH database were sorted into the following major categories: cancer 

bioassays, in vivo studies of cancer-related endpoints, and in vitro studies of cancer-related 

endpoints. These categories were further sorted by route (for bioassays) or by endpoint (for 
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cancer-related endpoints). Each study was reviewed, and critical study details were extracted 

into tables for each individual endpoint (see Section 4). The tables also include an initial 

determination of whether the data from each study meet selection criteria for use in the RPF 

analysis. Studies with data on selected PAHs and benzo[a]pyrene were considered for RPF 

determination, even if a particular PAH has not been classified by U.S. EPA or International 

Agency for Research on Cancer (IARC) as a carcinogen. Studies were included in the analysis if 

the following selection criteria were met: 

• Benzo[a]pyrene was tested simultaneously with another PAH; 

• A statistically increased incidence of tumors was observed with benzo[a]pyrene 

administration, compared with control incidence; 

• Benzo[a]pyrene produced a statistically significant change in a cancer-related endpoint 

finding; 

• Quantitative results were presented; 

• The carcinogenic response observed in either the benzo[a]pyrene- or other PAH-treated 

animals at the lowest dose level was not saturated (i.e., tumor incidence at the lowest 

dose was <90%), with the exception of tumor multiplicity findings; and 

• There were no study quality concerns or potential confounding factors that precluded use 

(e.g., no concurrent control, different vehicles, strains, etc. were used for the tested PAH 

and benzo[a]pyrene; use of cocarcinogenic vehicle; PAHs of questionable purity; 

unexplained mortality in treated or control animals). 

If the above criteria were met, studies were selected for use in the analysis regardless of 

whether positive or negative results were reported. Studies with positive findings were used for 

calculation of RPFs. Studies with negative findings were used in a weight of evidence 

evaluation of potential carcinogenicity (discussed in Section 6.1). 

Dose-response data were extracted from studies with positive findings that met selection 

criteria. For studies that reported results graphically, individual data points were extracted using 

digitizing software. In all, over 300 data sets were extracted, reflecting dose-response data from 

at least one study for 50 of the 74 PAHs included in the analysis. All of the extracted data are 

presented in Appendix C of this report. 

Statistical analyses were performed on tumor bioassay data to determine whether the 

tumor incidence or multiplicity observed at a particular dose represented a statistically significant 

increase over controls. If statistical analyses were not described in the original report, incidence 

data were analyzed using Fisher’s Exact test and the Cochran-Armitage trend test. Positive 
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findings were indicated by a significant (p < 0.05) difference for at least one dose group by 

comparison to control (in Fisher’s Exact or an equivalent test) or a significant dose-response 

trend (Cochran-Armitage or equivalent) for multi-dose studies. For tumor bioassay data reported 

as tumor count, a t-test was conducted (when variance data were available) to determine whether 

the count was significantly different from control (p < 0.05). The results of the statistical 

analyses are shown with the dose-response data in Appendix C. Statistical analyses of the 

cancer-related endpoint data were not conducted; the study author’s conclusions as to response 

(positive or negative) was used. 

Section 5 describes both the methods used for dose-response assessment and RPF 

calculation in detail. The general equation for estimating an RPF was the ratio of the slope of the 

dose-response curve for the subject PAH to the slope of the dose-response curve for 

benzo[a]pyrene. For bioassay data, tumor incidences were modeled using the multistage model 

within the U.S. EPA Benchmark Dose (BMD) Software (Version 1.3.2). For cancer-related 

endpoint data in quantal form, this model was also used; for continuous data (either tumor 

multiplicity or cancer-related endpoint data), the simplest continuous model (linear) within the 

software was applied. Whenever the data allowed, benchmark response (BMR) values of 10% 

for quantal data and 1 standard deviation from the control value for continuous data were used to 

calculate the slope by linear extrapolation to the origin for consistency across data sets. 

Alternative BMR values were used in select instances, as described in Section 5.3. For data sets 

that included only a single dose, or those for which no model fit was achieved with the selected 

models, a point estimate RPF was calculated. 

The RPFs calculated from individual studies for each PAH were used in a weight of 

evidence evaluation to assess the potential carcinogenicity of each compound (see Section 6) and 

in the derivation of a final RPF for each compound (Section 7). The selection of PAHs to be 

included in the RPF approach began with an evaluation of whether the available data were 

adequate to assess the potential carcinogenicity of each compound. At least one RPF value was 

calculated for each of 50 PAHs. For 16 of these compounds, only a single RPF value derived 

from an in vitro cancer-related endpoint (primarily mutagenicity assays) was available (see 

Table 6-1). Due to the limited data available for these 16 compounds, no further evaluation of 

these PAHs was conducted, and they were not selected for inclusion in the RPF approach. 

For the remaining 34 PAHs, a weight of evidence evaluation (see Figure 6-1) was 

conducted to assess the evidence that each PAH could induce a carcinogenic response. This 

evaluation did not constitute a formal weight of evidence evaluation of carcinogenic potential; 

rather, an expedited approach was developed using the data collected to determine whether the 

available information for each PAH was adequate to draw a conclusion regarding carcinogenic 

potential. When the data were considered adequate for a given PAH, it was selected for 
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inclusion in the RPF approach; if the data were not considered adequate to assess potential 

carcinogenicity, the PAH was excluded. In vivo tumor bioassays that included benzo[a]pyrene 

were given the greatest weight in assessing the potential carcinogenicity of a given PAH; data 

from other bioassays and cancer-related endpoint studies were used to supplement the weight of 

evidence when the bioassay data that included benzo[a]pyrene were conflicting or negative. 

Structural alerts for PAH carcinogenicity or mutagenicity (as defined in Section 2.5 as the 

presence of a classic bay region or fjord region in a PAH containing at least four benzene rings) 

were noted in the evaluation for each PAH, but were not used explicitly in the weight of 

evidence evaluation. 

The weight of evidence evaluation (Section 6) indicated that the available data were 

adequate to determine that 23 of the 34 PAHs were potentially carcinogenic, that three PAHs 

(anthracene, phenanthrene, and pyrene) exhibited little or no carcinogenic potential, and that data 

were inadequate to evaluate the carcinogenic potential for eight PAHs. The eight PAHs with 

inadequate data were excluded from the RPF approach. For the three PAHs for which there were 

sufficient data to conclude that the PAH had minimal carcinogenic potential (i.e., robust negative 

tumor bioassay data and cancer-related endpoint data), a final RPF of 0 was recommended. 

While there is little quantitative difference between selecting a final RPF of 0 for a given PAH 

and excluding that PAH from the RPF approach, this is an important distinction for uncertainty 

analysis. There is substantial uncertainty in the risk associated with PAHs that are excluded 

from the RPF approach due to inadequate data; these compounds could be of low or high 

potency. However, for PAHs with an RPF of 0, there is evidence to suggest that these 

compounds are of little or no carcinogenic potential, and the uncertainty associated with the 

cancer risk for these compounds is markedly reduced. 

For each of the remaining 23 compounds, a final nonzero RPF was derived. A number of 

options were considered for deriving an RPF from among the numerous values calculated for 

each individual PAH. These options included: prioritizing bioassay RPFs from different 

exposure routes based on relevance to environmentally-relevant routes; prioritizing bioassay 

RPFs based on target organs considered relevant to human susceptibility to PAH carcinogenesis; 

prioritizing RPFs based on quality of the underlying study; prioritizing cancer-related endpoints 

by their correlation with bioassay potency (i.e., ability to predict bioassay potency); and 

aggregating RPFs across all bioassays, all cancer-related endpoints, or across all endpoints. In 

the end, it was concluded that the available data did not provide a clear scientific basis for 

prioritizing RPFs except for a preference for bioassay data over cancer-related endpoints. As a 

consequence, final RPFs were derived from bioassay data for any PAH that had at least one RPF 

based on a bioassay. 
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For each potentially carcinogenic PAH with bioassay data, the average RPF was 

calculated from bioassays with positive results. For those PAHs that did not have any estimated 

RPF based on a bioassay, but for which the weight of evidence evaluation indicated a potential 

for carcinogenic response (e.g., dibenz[a,c]anthracene), the final RPF was calculated from all 

cancer-related endpoint studies with positive results. In both cases, nonpositive results were not 

included in the calculation. The final RPF for each PAH was reported to one significant figure. 

The range of RPF values was also reported. Presenting the RPFs in this manner provides an 

average and maximum estimate for each PAH that has data from multiple studies. 

All tumor bioassay RPFs (across all exposure routes, species, sexes, and including both 

tumor incidence and tumor multiplicity RPFs) were combined to estimate the mean and range, 

except as follows. In some cases, two separate RPFs were calculated in the same group of 

animals. There were two situations in which this occurred: RPFs for different target organs in 

the same animals, and RPFs based on incidence of tumors and tumor count in the same animals 

In these instances, the higher value of the two RPFs was included in the average and range, and 

the lower value was dropped from the combined data. 

Several options were considered for the determination of a final RPFs (e.g., arithmetic 

mean, geometric mean, weighted average, maximum, or order of magnitude estimates). The 

arithmetic mean and range were chosen as a simple approach to describing the calculated RPF 

values available for each PAH. Other statistical measures (i.e., geometric mean, weighted 

average) were not considered appropriate due to the limited number of RPF values calculated for 

most PAHs and the variability in the RPF estimates. Most PAHs (19/26, 73%) had ≤3 calculated 

RPF values and the range of RPF values was greater than an order of magnitude for several 

compounds (6/26 PAHs). The variability in RPF estimates is likely due to differences in study 

design parameters (e.g., route, species/strain, exposure duration, exposure during sensitive time 

periods, initiation vs. promotion and complete carcinogenesis protocols, tumor incidence vs. 

multiplicity reporting) and dose-response methods (modeled vs. point estimates). Calculation of 

a weighted average was not possible because there is no clear biological rationale for choosing 

among study types or tumor data outcomes. Providing order of magnitude estimates, as has been 

previously done for estimating RPFs for PAHs, was not considered to be superior to calculating 

simple means. Including the range in the estimated RPFs was considered to be informative to the 

user for characterizing uncertainty. 

Once a final RPF was derived for a given PAH, the resulting value was assigned a 

relative confidence rating of high, medium, or low confidence. The relative confidence rating 

characterized the nature of the database upon which the final RPF was based. Confidence 

rankings were based on the robustness of the database. For final RPFs based on tumor bioassay 

data, confidence ratings considered both the available tumor bioassays and the size and 
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consistency of the cancer-related endpoint database. The most important factors that were 

considered included the availability of in vivo data and whether multiple exposure routes were 

represented. Other database characteristics that were considered important included the strength 

of evidence of genotoxicity data and SAR information, the availability of more than one in vivo 

study, and whether effects were evident in more than one sex or species. Very low relative 

confidence was reserved for final RPFs based on cancer-related endpoint data only (e.g., 

dibenz[a,c]anthracene). An RPF of zero was only applied if the data implied high or medium 

relative confidence. 

Table 1 shows the average RPFs based on tumor bioassay data with their associated range 

and relative confidence ratings, and an overview of the tumor bioassay database (total number of 

studies, exposure routes tested, species tested, sexes tested) for each PAH. Table 2 shows the 

average RPF for dibenz[a,c]anthracene, the only RPF based on cancer-related endpoint data, 

with its associated range, relative confidence rating, and an overview of the database for this 

compound. 
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Table 1. PAHs with final RPFs based on tumor bioassay data 

PAH 

Average 

RPF 

Range of 

RPFs 

Relative 

confidence 

Number of 

datasets Exposure routes tested 

Species 

tested Sexes t

Anthanthrene 0.4 0.2–0.5 Medium 2 Dermal, lung implantation Mouse, rat F Anthracene 0 0 Medium 1 (Negative) Dermal Mouse F Benz[a]anthracene 0.2 0.02–0.4 Medium 3 Dermal, intraperitoneal Mouse F, M Benz[b,c]aceanthrylene, 11H- 0.05 0.05 Low 1 Dermal Mouse F Benzo[b]fluoranthene 0.5 0.1–2 High 5 Dermal, intraperitoneal, lung implantation Mouse, rat F, M Benz[e]aceanthrylene 0.9 0.5–1 Low 2 Dermal Mouse F, M Benzo[g,h,i]perylene 0.009 0.009 Low 1 Lung implantation Rat F Benz[j]aceanthrylene 60 60 Low 1 Intraperitoneal Mouse F Benzo[j]fluoranthene 0.3 0.01–1 High 5 Dermal, intraperitoneal, lung implantation Mouse, rat F, M Benzo[k]fluoranthene 0.03 0.03–0.03 Medium 2 Dermal, lung implantation Mouse, rat F Benz[l]aceanthrylene 5 4–7 Low 2 Dermal Mouse F, M Chrysene 0.1 0.04–0.2 High 7 Dermal, intraperitoneal, lung implantation Mouse, rat F, M Cyclopenta[c,d]pyrene 0.4 0.07–1 Medium 5 Dermal, intraperitoneal Mouse F, M Cyclopenta[d,e,f]chrysene, 4H- 0.3 0.2–0.5 Low 2 Dermal Mouse F Dibenzo[a,e]fluoranthene 0.9 0.7–1 Low 2 Dermal Mouse F Dibenzo[a,e]pyrene 0.4 0.3–0.4 Low 2 Dermal Mouse FDibenz[a,h]anthracene 6 1–10 High 3 Dermal, intraperitoneal, lung implantation Mouse, rat F, M Dibenzo[a,h]pyrene 0.9 0.9 Low 1 Dermal Mouse F Dibenzo[a,i]pyrene 0.6 0.5–0.7 Low 2 Dermal Mouse FDibenzo[a,l]pyrene 30 10–40 Medium 3 Dermal, intraperitoneal Mouse F, M Fluoranthene 0.08 0.009–0.2 Low 6 Intraperitoneal Mouse F, M Indeno[1,2,3-c,d]pyrene 0.07 0.07 Low 1 Lung implantation Rat F 

Naphtho[2,3-e]pyrene 0.3 0.3 Low 1 Dermal Mouse F 

Phenanthrene 0 0 High 3 (Negative) Dermal, intraperitoneal, lung implantation Mouse, rat F, M Pyrene 0 0 High 7 (Negative) Dermal, intraperitoneal Mouse F, M 

NA = not applicable; M = male; F = female 
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Table 2. PAHs with final RPFs based on cancer-related endpoint data 

(no tumor bioassay data available)

PAH 

Average 

RPF 

Range of 

RPFs 

Relative 

confidence Types of studies Multiple dose studies

Dibenz[a,c]anthracene 4 0.04–50 Very low Total = 14 studies

One in vivo DNA adduct 

Six in vitro bacterial 

mutagenicity 

One in vitro mammalian 

mutagenicity 

One in vitro 

morphological/malignant 

transformation 

Three in vitro DNA 

damage 

Two in vitro DNA adducts

Total = 6 studies

Four in vitro bacterial 

mutagenicity 

One in vitro DNA 

damage 

One in vitro DNA 

adduct 

According to the Supplemental Guidance for Assessing Susceptibility from Early-Life 

Exposure to Carcinogens (U.S. EPA, 2005b), benzo[a]pyrene is carcinogenic by a mutagenic 

mode of action. The PAH compounds for which a RPF value was derived are also considered to 

be carcinogenic by a mutagenic mode of action (see Section 2.4 for discussion of similarities in 

mode of action for PAHs). When assessing PAH cancer risks for life-stages under 16 years of 

age, or for lifetime exposures that include early-life exposures, the RPF values should be applied 

with specific exposure information to the benzo[a]pyrene cancer risk estimates including 

adjustment for early-life susceptibility, through the application of age-dependent adjustment 

factors (ADAFs). 

A description of uncertainties and limitations is crucial to interpretation of the RPF 

approach for PAH mixtures risk assessment (see Section 8). Many of the general uncertainties 

related to chemical-specific risk assessment are also applicable to the proposed RPF approach for 

PAHs (e.g., appropriateness of animal models, low-dose and interspecies extrapolation, 

variability within the human population). Use of a component-based approach for mixtures risk 

assessment leads to additional uncertainties related to adequate characterization of the mixture 

and the potential interactions that may occur between individual components within the mixture 

(i.e., PAHs and other chemicals). The RPF approach is limited by the small number of PAHs for 

which there are analytical chemistry and toxicology data, and thus may result in underestimation 

of actual cancer risks from complex PAH mixtures. There are uncertainties and limitations 

related to the size and nature of the PAH database, the human relevance of animal data, 

assumptions regarding mode of action and dose additivity, and cross-route extrapolation. 

Specific uncertainties that are related to dose-response assessment (i.e., calculation of RPFs) and 

the selection of single RPF values for each PAH are also discussed in Section 8. 

In summary, the current analysis represents a significant improvement upon the previous 

component-based approaches for PAH mixtures risk assessment. One of the most important 
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improvements is the consideration of data from a comprehensive review of the scientific 

literature dating from the 1950s through 2009 on the carcinogenicity and genotoxicity of PAHs. 

The search identified over 900 individual publications for a target list of 74 PAHs that have been 

identified in environmental media and for which toxicological data are available. Review of 

these publications resulted in the identification of more than 600 papers that included 

carcinogenicity or cancer-related endpoint data on at least one PAH and benzo[a]pyrene tested at 

the same time. Dose-response data were extracted, and RPFs from individual studies were 

calculated from over 300 data sets representing 50 individual PAHs. A weight of evidence 

evaluation was conducted to evaluate the evidence for potential carcinogenicity of 34 of these 

PAHs; data were inadequate to conduct such an evaluation for the remaining 16 compounds. A 

final RPF was derived for each PAH based on tumor bioassay data (if available) or cancerrelated endpoint data (if no tumor bioassay RPFs were available). Final RPFs were derived for 

26 PAHs, significantly increasing the number of PAHs that can be addressed through this 

approach. Each RPF was assigned a relative confidence rating reflecting the nature of the tumor 

bioassay or cancer-related endpoint database that was used to derive the final RPF for that PAH. 
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CONTENTS 

LIST OF TABLES ..................................................................................................................... xvii

LIST OF FIGURES........................................................................................................................ 1

LIST OF ABBREVIATIONS AND ACRONYMS....................................................................... 4

AUTHORS, CONTRIBUTORS, AND REVIEWERS.................................................................. 5

1. BACKGROUND FOR THE DEVELOPMENT OF A PAH MIXTURES HEALTH 

ASSESSMENT.......................................................................................................................... 7

2. RATIONALE FOR RECOMMENDING AN RPF APPROACH ............................................ 8

2.1. PAHs AS A CHEMICAL CLASS .................................................................................... 11

2.2. THE TOXICOLOGICAL DATABASE FOR PAHs ........................................................ 27

2.3. BENZO[A]PYRENE AS AN INDEX CHEMICAL......................................................... 28

2.4. SIMILARITIES IN MODE OF CARCINOGENIC ACTION FOR PAHs ...................... 31

2.5. STRUCTURAL ALERTS FOR PAH CARCINOGENESIS............................................ 42

2.6. SIMILARITIES IN RELATIVE POTENCY ACROSS ENDPOINTS............................ 43

2.7. SIMILARITIES IN RELATIVE POTENCY ESTIMATES ACROSS SPECIES 

AND EXPOSURE ROUTES............................................................................................ 45

2.8. DOSE ADDITIVITY OF PAHs IN COMBINED EXPOSURES .................................... 46

3. DISCUSSION OF PREVIOUSLY PUBLISHED RPF APPROACHES ............................... 50

3.1. PREVIOUS EFFORTS TO VALIDATE RPF APPROACH ........................................... 58

4. EVALUATION OF THE CARCINOGENICITY OF INDIVIDUAL PAHs......................... 60

4.1. DATABASE OF STUDIES ON PAH CARCINOGENICITY AND CANCERRELATED ENDPOINTS ................................................................................................. 60

4.2. STUDIES IN HUMANS ................................................................................................... 62

4.3. STUDIES IN ANIMALS .................................................................................................. 62

4.3.1. In Vivo Cancer Bioassays in Animals ......................................................................... 89

4.3.1.1. Dermal Exposure ................................................................................................ 89

4.3.1.2. Intraperitoneal Exposure..................................................................................... 92

4.3.1.3. Subcutaneous Injection Exposure....................................................................... 93

4.3.1.4. Oral Exposure ..................................................................................................... 94

4.3.1.5. Other Routes ....................................................................................................... 95

4.3.2. In Vivo Studies of Cancer-Related Endpoints............................................................. 95

4.3.2.1. DNA Adducts...................................................................................................... 96

4.3.2.2. Clastogenicity or Sister Chromatid Exchange Frequency .................................. 98

4.3.2.3. In Vivo Mutagenicity.......................................................................................... 99

4.3.3. In Vitro Studies of Cancer-Related Endpoints........................................................... 100

4.3.3.1. Bacterial Mutagenicity...................................................................................... 100

4.3.3.2. Mammalian Mutagenicity................................................................................. 101

4.3.3.3. Morphological/Malignant Cell Transformation................................................ 102

4.3.3.4. DNA Adducts.................................................................................................... 103

4.3.3.5. DNA Damage/Repair........................................................................................ 104

4.3.3.6. Clastogenicity or Sister Chromatid Exchange Frequency ................................ 104

4.4. SUMMARY OF INFORMATION AVAILABLE TO DEVELOP RPFS FOR 

INDIVIDUAL PAHs ...................................................................................................... 106
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5. METHODS FOR DOSE-RESPONSE ASSESSMENT AND RPF CALCULATION ........ 106

5.1. CHOICE OF DOSE-RESPONSE DATA ....................................................................... 106

5.2. OVERALL FORM OF RPF ESTIMATE ....................................................................... 108

5.3. RPF CALCULATION FOR MULTIDOSE DATASETS .............................................. 108

5.4. RPF CALCULATION FOR SINGLE DOSE DATASETS............................................ 110

5.5. DOSE CONVERSION FOR RPF CALCULATION...................................................... 111

5.6. SPECIAL CONSIDERATIONS FOR RPF CALCULATION USING TUMOR 

BIOASSAY DATA ........................................................................................................ 112

5.7. SPECIAL CONSIDERATIONS FOR RPF CALCULATION USING CANCERRELATED ENDPOINT DATA ..................................................................................... 112

6. SELECTION OF PAHS FOR INCLUSION IN RELATIVE POTENCY APPROACH ..... 113

6.1. METHOD FOR SELECTING PAHS FOR INCLUSION IN RELATIVE 

POTENCY APPROACH................................................................................................ 115

6.2. WEIGHT OF EVIDENCE EVALUATION FOR 34 INDIVIDUAL PAHS.................. 118

7. CONCLUSIONS ................................................................................................................... 189

7.1. METHODS FOR DERIVING FINAL RPFs .................................................................. 189

7.2. CONFIDENCE RATINGS FOR FINAL RPFs .............................................................. 191

7.3. SUSCEPTIBILITY FROM EARLY LIFE EXPOSURE TO CARCINOGENS ............ 196

8. UNCERTAINTIES ASSOCIATED WITH RPF APPROACHES ....................................... 197

8.1. UNCERTAINTY IN DOSE-RESPONSE ASSESSMENT FOR INDIVIDUAL 

PAHS .............................................................................................................................. 198

8.2. UNCERTAINTY IN SELECTING PAHs FOR INCLUSION IN RPF 

APPROACH ................................................................................................................... 200

8.3. UNCERTAINTY IN DERIVING A FINAL RPF FOR EACH PAH............................. 202

8.4. UNCERTAINTY IN USE OF ANIMAL DATA TO PREDICT HUMAN 

CANCER RISK .............................................................................................................. 206

8.5. UNCERTAINTY IN THE ASSUMPTIONS OF COMMON MODE OF ACTION 

AND DOSE ADDITIVITY ............................................................................................ 208

8.6. UNCERTAINTY IN EXTRAPOLATING RPFs ACROSS EXPOSURE ROUTES..... 209

9. REFERENCES...................................................................................................................... 215

APPENDIX A. SECONDARY SOURCES REVIEWED FOR IDENTIFICATION OF 

PRIMARY LITERATURE....................................................................................................... 1

APPENDIX B. BIBLIOGRAPHY OF STUDIES WITHOUT BENZO[A]PYRENE AS A 

REFERENCE COMPOUND................................................................................................ B-1

B.1. BIBLIOGRAPHY OF BIOASSAYS WITHOUT BENZO[A]PYRENE ...................... B-4

B.2. BIBLIOGRAPHY OF STUDIES ON CANCER-RELATED ENDPOINTS 

WITHOUT BENZO[A]PYRENE ................................................................................ B-10

APPENDIX C. DOSE-RESPONSE DATA FOR POTENCY CALCULATIONS.................. C-1
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APPENDIX D. BENCHMARK DOSE MODELING OUTPUTS ........................................... D-1

D.1. DERMAL BIOASSAYS ................................................................................................ D-1

D.2. INTRAPERITONEAL BIOASSAYS .......................................................................... D-45

D.3. LUNG IMPLANTATION BIOASSAYS..................................................................... D-67

D.4. BACTERIAL MUTAGENICITY ................................................................................ D-87

D.5. MAMMALIAN MUTAGENICITY............................................................................. D-96

D.6. MALIGNANT TRANSFORMATION ...................................................................... D-124

D.7. IN VITRO DNA DAMAGE....................................................................................... D-156

APPENDIX E. CALCULATION OF RPFs...............................................................................E-1

APPENDIX F. EXAMPLE CALCULATION OF RPF DETECTION LIMIT.........................F-1

APPENDIX G: EVALUATION OF ALTERNATIVES FOR RANKING RPFs ..................... G-1

G.1. OPTIONS FOR RANKING TUMOR BIOASSAY RPFS ............................................ G-1

G.2. RANKING NON-BIOASSAY DATA........................................................................... G-6
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LIST OF TABLES 

2-1. PAHs evaluated in the RPF analysis .................................................................................... 12 

2-2. Studies of binary mixtures of PAHs and tumorigenicity...................................................... 47 

3-1. Comparison among various relative potency estimates for PAHs from the published 

literature and regulatory agencies (1984–2004)................................................................... 51 

4-1. Study summaries: dermal bioassays of benzo[a]pyrene and at least one other PAH.......... 64 

4-2. Study summaries: intraperitoneal bioassays of benzo[a]pyrene and at least one 

other PAH............................................................................................................................. 68 

4-3. Study summaries: subcutaneous bioassays of benzo[a]pyrene and at least one 

other PAH............................................................................................................................. 69 

4-4. Study summaries: oral bioassays of benzo[a]pyrene and at least one other PAH............... 70 

4-5. Study summaries: other route bioassays of benzo[a]pyrene and at least one other 

PAH...................................................................................................................................... 71 

4-6. Study summaries: in vivo DNA adducts with benzo[a]pyrene and at least one 

other PAH............................................................................................................................. 72 

4-7. Study summaries: in vivo clastogenicity or sister chromatid exchange with 

benzo[a]pyrene and at least one other PAH ......................................................................... 74 

4-8. Study summaries: in vivo mutagenicity with benzo[a]pyrene and at least one 

other PAH............................................................................................................................. 75 

4-9. Study summaries: in vitro bacterial mutagenicity with benzo[a]pyrene and at least 

one other PAH...................................................................................................................... 76 

4-10. Study summaries: in vitro mammalian mutagenicity assays with benzo[a]pyrene and 

at least one other PAH........................................................................................................ 79 

4-11. Study summaries: in vitro morphological/malignant cell transformation with 

benzo[a]pyrene and at least one other PAH ....................................................................... 82 

4-12. Study summaries: in vitro DNA adducts with benzo[a]pyrene and at least one 

other PAH........................................................................................................................... 84 

4-13. Study summaries: in vitro DNA damage, repair, or synthesis with benzo[a]pyrene 

and at least one other PAH................................................................................................. 85 

4-14. Study summaries: in vitro clastogenicity or sister chromatid exchange with 

benzo[a]pyrene and at least one other PAH ....................................................................... 87 
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5-1 Comparison between molar and mass-based RPF............................................................... 111 

6-1. PAHs with only one RPF from in vitro cancer-related endpoint study and excluded 

from RPF approach ............................................................................................................ 114 

6-2. Results of weight of evidence evaluation of 34 PAHs..........Error! Bookmark not defined.

7-1. Final RPFs based on tumor bioassay data .......................................................................... 193 

7-2. Final RPFs based on cancer-related endpoint data (no tumor bioassay 

data available) .................................................................................................................... 195 

8-1. Results of simple linear regression of log–transformed average genotoxicity RPF vs. 

log average tumor bioassay RPF........................................................................................ 204 

8-2. PAHs with RPFs of varying confidence............................................................................. 205 

8-3. Comparisons among average tumor bioassay RPF values by exposure route and 

target organ......................................................................................................................... 211 

B-1. Bioassays with and without benzo[a]pyrene by PAH........................................................ B-2 

C-1. Dermal bioassays: dose response information for incidence data ..................................... C-2 

C-2. Dermal bioassays: dose-response information for tumor multiplicity............................... C-8 

C-3. Intraperitoneal bioassays: dose response information for incidence data........................ C-13 

C-4. Intraperitoneal bioassays: dose-response information for tumor multiplicity ................. C-22

C-5. Lung implantation bioassays: dose response information for incidence data.................. C-26 

C-6. In vitro bacterial mutagenicity: data use.......................................................................... C-30 

C-7. In vitro bacterial mutagenicity: dose response data......................................................... C-33 

C-8. In vitro mammalian mutagenicity: data use..................................................................... C-51 

C-9. In vitro mammalian mutagenicity: dose response data.................................................... C-53 

C-10. In vitro malignant/morphological cell transformation: data use.................................... C-61 

C-11. In vitro malignant/morphological cell transformation: dose response data................... C-63

C-12. In vitro DNA adducts: data use...................................................................................... C-71 

C-13. In vitro DNA adducts: dose response data..................................................................... C-72 
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C-14. In vitro DNA damage: data use ..................................................................................... C-75 

C-15. In vitro DNA damage: dose response data .................................................................... C-76 

C-16. In vitro clastogenicity: data use ..................................................................................... C-80 

C-17. In vitro clastogenicity: dose response data .................................................................... C-81 

C-18. In vivo DNA adducts: data use ...................................................................................... C-82 

C-19. In vivo DNA adducts: dose response data ..................................................................... C-83 

C-20. In vivo clastogenicity: data use...................................................................................... C-90 

E-1. Dermal bioassays: RPF calculations for incidence data .....................................................E-2 

E-2. Dermal bioassays: RPF calculations for multiplicity data ..................................................E-5 

E-3. Intraperitoneal bioassays: RPF calculations for incidence data.........................................E-7 

E-4. Intraperitoneal bioassays: RPF calculations for multiplicity data......................................E-9 

E-5. Lung implantation bioassays: RPF calculations (incidence data)....................................E-10 

E-6. In vivo DNA adducts: RPF calculations ...........................................................................E-11 

E-7. In vivo clastogenicity or sister chromatid exchange: RPF calculation ............................E-13 

E-8. In vitro bacterial mutagenicity: RPF calculations............................................................E-15 

E-9. In vitro mammalian mutagenicity: RPF calculations.......................................................E-21 

E-10. In vitro morphological/malignant transformation: RPF calculation ..............................E-24 

E-11. In vitro DNA adducts: RPF calculations.........................................................................E-27 

E-12. In vitro DNA damage: RPF calculations .......................................................................E-29 

E-13. In vitro clastogenicity or sister chromatid exchange: RPF calculations .........................E-31 

F-1. Example data for calculation of RPF detection limit ..........................................................F-1 

G-1. Average RPF value by exposure route .............................................................................. G-3 

G-2. Comparisons among average tumor bioassay-based RPF values by data availability 

or calculation method ........................................................................................................ G-5 

G-3. Correlation between LED and TD25 by endpoint .............................................................. G-7 
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G-4. Correlation between tumor potency (log 1/TD50) and mutagenic potency........................ G-7 

G-5. Results of simple linear regression of log–transformed average genotoxicity RPF vs. 

log average tumor bioassay RPF....................................................................................... G-9 
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1 LIST OF FIGURES 

2 

3 

4 2-1. Structural features of PAHs.................................................................................................. 27 

5 

6 2-2. Metabolic pathways for benzo[a]pyrene .............................................................................. 32 

7 

8 2-3. Overview of the proposed key events in the mode of action for PAH 

9 carcinogenicity. .................................................................................................................... 34 

10 

2-4. Structures of the four stereoisomeric adduct moieties, anti-[BaP]-N

2

11 -dG, derived from 

12 the trans- or cis-covalent binding of (+)-anti-BaP diol epoxide or (-)-anti-BaPdiol 

13 epoxide to dG residues in DNA. .......................................................................................... 35 

14 

15 2-5. Depurinating adducts of benzo[a]pyrene formed by one-electron oxidation....................... 36

16 

17 2-6. Spectrum of DNA adducts anticipated with PAH o-quinones. ............................................ 37 

18 

19 2-7. Interaction of PAHs with the Ah receptor – regulation of genes related to induction of 

20 metabolism and cell differentiation and proliferation. ......................................................... 39 

21 

22 6-1. Weight of evidence assessment of potential carcinogenicity. ............................................ 116 

23 

24 6-2. 2,3-Acepyrene (ACEP) RPFs............................................................................................. 121 

25 

26 6-3. Anthanthrene (AA) RPFs. .................................................................................................. 123 

27 

28 6-4. Anthracene (AC) RPFs....................................................................................................... 125 

29 

30 6-5. Benz[a]anthracene (BaA) RPFs.......................................................................................... 127 

31 

32 6-6. Benz[b,c]aceanthrylene (BbcAC) RPFs............................................................................. 129 

33 

34 6-7. Benzo[b]fluoranthene (BbF) RPFs..................................................................................... 131 

35 

36 6-8. 11H-Benzo[b]fluorene (BbFE) RPFs. ................................................................................ 133 

37 

38 6-9. Benz[e]aceanthrylene (BeAC) RPFs.................................................................................. 135 

39 

40 6-10. Benzo[e]pyrene (BeP) RPFs............................................................................................. 137 

41 

42 6-11. Benzo[g,h,i]fluoranthene (BghiF) RPFs........................................................................... 139 

43 

44 6-12. Benzo[g,h,i]perylene (BghiP) RPFs. ................................................................................ 141 

45 

46 6-13. Benz[j]aceanthrylene (BjAC) RPFs ................................................................................. 143 

47 

48 6-14. Benzo[j]fluoranthene (BjF) RPFs..................................................................................... 145 

49 
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1 6-15. Benzo[k]fluoranthene (BkF) RPFs................................................................................... 147 

2 

3 6-16. Benz[l]aceanthrylene (BlAC) RPFs. ................................................................................ 149 

4 

5 6-17. Chrysene (CH) RPFs. ....................................................................................................... 151 

6 

7 6-18. Coronene (CO) RPFs........................................................................................................ 153 

8 

9 6-19. Cyclopenta[c,d]pyrene (CPcdP) RPFs. ............................................................................ 155 

10 

11 6-20. Cyclopenta[d,e,f]chrysene (CPdefC) RPFs...................................................................... 157 

12 

13 6-21. Dibenz[a,c]anthracene (DBacA) RPFs............................................................................. 159 

14 

15 6-22. Dibenzo[a,e]fluoranthene (DBaeF) RPFs......................................................................... 161 

16 

17 6-23. Dibenzo[a,e]pyrene (DBaeP) RPFs.................................................................................. 163 

18 

19 6-24. Dibenz[a,h]anthracene (DBahA) RPFs. ........................................................................... 165 

20 

21 6-25. Dibenzo[a,h]pyrene (DBahP) RPFs. ................................................................................ 167 

22 

23 6-26. Dibenzo[a,i]pyrene (DbaiP) RPFs.................................................................................... 169 

24 

25 6-27. Dibenzo[a,l]pyrene (DBalP) RPFs. .................................................................................. 172 

26 

27 6-28. Fluoranthene (FA) RPFs................................................................................................... 174 

28 

29 6-29. Fluorene (FE) RPFs.......................................................................................................... 176 

30 

31 6-30. Indeno[1,2,3-c,d]pyrene (IP) RPFs................................................................................... 178 

32 

33 6-31. Naphtho[2,3-e]pyrene (N23eP) RPFs............................................................................... 180 

34 

35 6-32. Perylene (Pery) RPFs........................................................................................................ 182 

36 

37 6-33. Phenanthrene (PH) RPFs.................................................................................................. 184 

38 

39 6-34. Pyrene (Pyr) RPFs. ........................................................................................................... 186 

40 

41 6-35. Triphenylene (Tphen) RPFs ............................................................................................. 188 

42 

43 8-1. Correlation between incidence and multiplicity RPFs ....................................................... 203 

44 

45 G-1. Average bioassay RPF vs. average in vivo DNA adduct RPF ........................................ G-10 

46 

47 G-2. Average bioassay RPF vs. average in vivo nonbioassay RPF......................................... G-10 

48 

49 G-3. Average bioassay RPF vs. average nonbioassay RPF..................................................... G-11 
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1 

2 G-4. Average bioassay RPF vs. average in vitro non-bioassay RPF ....................................... G-11 

3 
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LIST OF ABBREVIATIONS AND ACRONYMS*1 

2 

3 

4 AEL acceptable exposure level 

5 Ah aryl hydrocarbon 

6 AhR Ah receptor 

7 AhREDNA Ah-responsive elements of DNA 

8 ARNT Ah-receptor nuclear translocator 

9 ATSDR Agency for Toxic Substances and Disease Registry 

10 AUC area under the curve 

11 BMD benchmark dose 

12 BMR benchmark response 

13 CASRN Chemical Abstract Service Registry Number 

14 CHO Chinese hamster ovary 

15 CYP cytochrome P450 

16 dG deoxyguanosine 

17 EAL environmental assessment level 

18 EOPP estimated order of potential potency 

19 EROD ethoxyresorufin O-deethylase 

20 HPRT hypoxanthine-guanine phosphoribosyl transferase gene 

21 Hsp90 heat shock protein 90 

22 IARC International Agency for Research on Cancer 

23 IRIS Integrated Risk Information System

24 LED lowest effective dose 

25 MGP manufactured gas plant 

26 MVK Moolgavkar-Venson-Knudsen two-stage model 

27 OEHHA Office of Environmental Health Hazard Assessment, California EPA 

28 PAC polycyclic aromatic compound 

29 PAH polycyclic aromatic hydrocarbon 

30 PCR polymerase chain reaction 

31 PEF potency equivalency factor 

32 RTECS Registry of Toxic Effects of Chemical Substances 

33 RPF relative potency factor 

34 RTD relative tumor dose 

35 SMART somatic mutation and recombination test

36 TK thymidine kinase locus 

37 TIDAL time-integrated DNA adduct level 

38 TEF toxicity equivalency factor 

39 TPA 12-O-tetra-decanoylphorbol-13-acetate 

40 U.S. EPA U.S. Environmental Protection Agency 

41 WHO World Health Organization 

42 

*

43 Abbreviations for PAH chemical names are provided in Table 2-1. 
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1 AUTHORS, CONTRIBUTORS, AND REVIEWERS 

2 

3 CHEMICAL MANAGERS

4 

5 Lynn Flowers, Ph.D., DABT 

6 National Center for Environmental Assessment 

7 Office of Research and Development 

8 U.S. Environmental Protection Agency 

9 Washington, DC 

10 

11 Martin Gehlhaus, III 

12 National Center for Environmental Assessment, IRIS Program 

13 Office of Research and Development 

14 U.S. Environmental Protection Agency 

15 Washington, DC 

16 

17 AUTHORS 

18 

19 Lynn Flowers, Ph.D., DABT 

20 National Center for Environmental Assessment 

21 Office of Research and Development 

22 U.S. Environmental Protection Agency 

23 Washington, DC 

24 

25 Martin Gehlhaus, III 

26 National Center for Environmental Assessment, IRIS Program 

27 Office of Research and Development 

28 U.S. Environmental Protection Agency 

29 Washington, DC 

30 

31 Karen Hogan 

32 National Center for Environmental Assessment, IRIS Program 

33 Office of Research and Development 

34 U.S. Environmental Protection Agency 

35 Washington, DC 

36 

37 Channa Keshava, Ph.D. 

38 National Center for Environmental Assessment, IRIS Program 

39 Office of Research and Development 

40 U.S. Environmental Protection Agency 

41 Washington, DC 

42 

43 Glenn Rice, Ph.D. 

44 National Center for Environmental Assessment 

45 Office of Research and Development 

46 U.S. Environmental Protection Agency 

47 Cincinnati, OH 

48 
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1 Jamie Strong, Ph.D. 

2 National Center for Environmental Assessment, IRIS Program 

3 Office of Research and Development 

4 U.S. Environmental Protection Agency 

5 Washington, DC 

6 

7 Linda Teuschler, Ph.D. 

8 National Center for Environmental Assessment 

9 Office of Research and Development 

10 U.S. Environmental Protection Agency 

11 Cincinnati, OH 

12 

13 Stephen Nesnow, Ph.D. 

14 Environmental Carcinogenesis Division 

15 National Health and Environmental Effects Research Laboratory 

16 Office of Research and Development 

17 Research Triangle Park, NC 

18 

19 Chao Chen, Ph.D. 

20 National Center for Environmental Assessment 

21 Office of Research and Development 

22 Washington, DC 

23 

24 Heather Carlson-Lynch, S.M. 

25 Syracuse Research Corporation, Inc. 

26 Syracuse, NY 

27 

28 Julie Stickney, Ph.D., DABT 

29 Syracuse Research Corporation, Inc. 

30 Syracuse, NY 

31 

32 Peter R. McClure, Ph.D., DABT 

33 Syracuse Research Corporation, Inc. 

34 Syracuse, NY 

35 

36 Amber Bacom 

37 Syracuse Research Corporation, Inc. 

38 Syracuse, NY 

39 

40 
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1 1. BACKGROUND FOR THE DEVELOPMENT OF A PAH MIXTURES HEALTH 

2 ASSESSMENT 

3 

4 

5 The U.S. Environmental Protection Agency (U.S. EPA) IRIS Program is undertaking a 

6 project to develop a PAH Mixtures Health Assessment. This assessment focuses on the RPF 

7 approach which is based on component PAHs in the mixture. In preparation for the development 

8 of the PAH Mixtures Health Assessment, U.S. EPA held a peer consultation workshop to outline 

9 some of the important issues. These issues are discussed in Peer Consultation Workshop on 

10 Approaches to Polycyclic Aromatic Hydrocarbon (PAH) Health Assessment (U.S. EPA, 2002) 

11 and the accompanying discussion document. 

12 Health assessments for 15 unsubstituted, nonheterocyclic polycyclic aromatic 

13 hydrocarbons (PAHs) with three or more rings are currently entered on EPA’s Integrated Risk 

14 Information System (IRIS) database. Benzo[a]pyrene is the only PAH for which quantitative 

15 oral, dermal, and inhalation data are available. 

16 In 1993, U.S. EPA published the Provisional Guidance for Quantitative Risk Assessment 

17 of PAHs (Provisional Guidance). The Provisional Guidance recommended estimated orders of 

18 potential potency (EOPP) for individual PAHs that could be used in a component-based 

19 approach to PAH mixtures risk assessment. The Provisional Guidance recommended EOPPs for 

20 7 PAHs categorized as Group B2 (probable human carcinogens) under 1986 U.S. EPA Cancer 

21 Guidelines: benzo[a]pyrene, benz[a]anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene, 

22 chrysene, dibenz[a,h]anthracene, and indeno[1,2,3-c,d]pyrene (U.S. EPA, 1993). The current 

23 analysis extends the 1993 Provisional Guidance and provides recommendations for further 

24 development of this approach to PAH mixtures risk assessment. The assessment includes the 

25 following: 

26 

27 (1) A rationale for recommending an order of potency, or relative potency factor (RPF), 

28 approach; 

29 

30 (2) A summary of previous approaches for developing the RPF approach for PAHs; 

31 

32 (3) Identification of individual carcinogenic PAHs that could be included in the RPF 

33 approach; 

34 

35 (4) Identification of potential index chemicals; 

36 

37 (5) Presentation of the available literature for in vivo carcinogenicity and both in vivo and in 

38 vitro cancer-related endpoint assays for individual PAHs; 

39 

40 (6) Development of a recommendation for the RPF approach for PAH mixtures; and 

41 
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1 (7) Characterization of strengths, weaknesses, and uncertainties associated with the 

2 recommended approaches. 

3 

4 

5 2. RATIONALE FOR RECOMMENDING AN RPF APPROACH 

6 

7 

8 PAHs are a concern as human health hazards, because many PAHs are demonstrated 

9 tumorigenic agents in animal bioassays and are active in in vivo or in vitro tests for genotoxicity 

10 or DNA damage. PAHs do not occur in the environment as isolated entities; they primarily 

11 occur in complex mixtures generated from the combustion or pyrolysis of substances containing 

12 carbon and hydrogen. Several complex mixtures of PAHs have been classified as possibly 

13 carcinogenic, probably carcinogenic, or carcinogenic to humans (Straif et al., 2005; U.S. EPA, 

14 2002; Bostrom et al., 2002; WHO, 1998; ATSDR, 1995; IARC, 1985, 1984a, b, 1983). 

15 In concordance with U.S. EPA (2000, 1986) guidance for health risk assessment of 

16 chemical mixtures, assessment of the cancer risk from long-term human exposure to a particular 

17 PAH mixture would best be conducted with quantitative information on the dose-response 

18 relationship for cancer from chronic exposure to the mixture of concern. When data for the 

19 mixture of concern are not available, U.S. EPA (2000, 1986) guidance recommends using 

20 toxicity data on a “sufficiently similar” mixture. However, quantitative cancer dose-response 

21 information exists only for a few complex mixtures generated from the combustion or pyrolysis 

22 of organic matter; for example, tobacco smoke, coke oven emissions, and emissions from roofing 

23 tar pots (see Bostrom et al., 2002; Albert et al., 1983). U.S. EPA’s IRIS database currently 

24 includes assessments for only three PAH-containing mixtures: coke oven emissions, creosote, 

25 and diesel emissions. The availability of oral carcinogenicity bioassays of manufactured gas 

26 plant (MGP) residue (Weyand et al., 1995) and coal tar preparations (Culp et al., 1998; Gaylor et 

27 al., 1998) has expanded the PAH mixture cancer database. 

28 Component-based approaches, involving an analysis of the toxicity of components of the 

29 mixture, are recommended when appropriate toxicity data on a complex mixture of concern, or 

30 on a “sufficiently similar” mixture, are unavailable (U.S. EPA, 2000, 1986). Component-based 

31 approaches involving dose addition (such as the RPF approach) are recommended when 

32 components in the mixture are judged to act in a toxicologically similar manner. In the RPF 

33 approach, doses of component chemicals that act in a toxicologically similar manner are added 

34 together, after scaling the doses relative to the potency of an index chemical (U.S. EPA, 2000, 

35 1986). Then, using the dose-response curve of the index chemical, the response to the total 

36 equivalent dose in the mixture is estimated. The index compound is typically the best-studied 

37 member of the class with the largest body of available data describing exposure and health 

38 effects. The index chemical should have a quantitative dose-response assessment of acceptable 
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1 scientific quality and must have (or be expected to have) similar toxic effects to the rest of the 

2 members of the class. 

3 For chemicals that act independently (e.g., by different modes of action), U.S. EPA 

4 guidance (2000, 1986) recommends a component-based approach involving response addition. 

5 In this approach, response is defined as the percentage or fraction of exposed individuals that 

6 show the effect of concern (i.e., the risk of having the effect). To apply a response-addition 

7 approach to a complex mixture, information on the dose-response relationships for the effect of 

8 concern from exposure to individual components must be available. Based on an analysis of the 

9 amount of each component in the mixture and its dose-response relationship, risks from exposure 

10 to the individual components are calculated and added together to estimate the risk for the effect 

11 from exposure to the complex mixture. 

12 Component-based approaches, either involving dose addition or response addition, 

13 include a general assumption that interaction effects at low dose levels either do not occur or are 

14 small enough to be neglected (U.S. EPA, 2000, 1986). However, when information on 

15 interactions among the components is available, U.S. EPA guidance recommends incorporating 

16 this information into the risk assessment, either as a part of a quantitative approach or as a 

17 qualitative evaluation. 

18 The assessment of cancer risk from chronic oral or inhalation exposure to complex PAH 

19 mixtures using component-based approaches is restricted by the limited availability of cancer 

20 dose-response data for individual PAHs. Benzo[a]pyrene is the only PAH that has dose-response 

21 data for cancer from chronic oral, inhalation or dermal exposure (WHO, 1998; ATSDR, 1995). 

22 The IRIS Program developed cancer assessments for 15 PAHs in the early 1990s, but a 

23 quantitative cancer assessment was only developed for benzo[a]pyrene. Six other PAHs were 

24 qualitatively assessed as B2, probable human carcinogens, but the available data were 

25 characterized as inadequate to develop oral or inhalation risk estimates. Thus, data are 

26 insufficient to use a component-based approach involving response addition for assessing cancer 

27 risks from PAH mixtures. Although a response addition approach does not require an 

28 assumption of similar toxicological action, it does require dose-response data for individual 

29 components. 

30 For exposure situations in which dose-response data for the PAH mixture or a sufficiently 

31 similar mixture are not available (e.g., the source of the PAH contamination may be mixed or 

32 unknown), there are at least three practical advantages of an RPF approach that uses 

33 benzo[a]pyrene as the index PAH: 

34 

35 (1) Benzo[a]pyrene is routinely assayed and detected in environmental media contaminated 

36 with PAH mixtures; 

37 

38 (2) Benzo[a]pyrene is the only PAH for which cancer dose-response data involving chronic 

39 exposures are available; and 
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1 

2 (3) There is a large database of studies in which the potency of benzo[a]pyrene is compared 

3 with the potency of other PAHs in various assays. 

4 

The database includes animal tumorigenicity15 assays involving dermal or parenteral 

6 administration, and in vivo and in vitro assays of cancer-related endpoints (e.g., various 

7 genotoxic endpoints). Thus, RPFs for a number of PAHs can be derived. 

8 The RPF approach involves two key assumptions: (1) the assumption of similar 

9 toxicological action as required by dose addition; and (2) the assumption that interactions among 

10 PAH mixture components do not occur at low levels of exposure typically encountered in the 

11 environment. 

12 Mechanistic studies indicate that the mutagenic and tumor-initiating activity of most 

13 carcinogenic PAHs requires metabolic activation to reactive intermediates (e.g., stereospecific 

14 dihydrodiol epoxides). For several PAHs, (e.g., benzo[a]pyrene, dibenz[a,h]anthracene, 

15 dibenzo[a,l]pyrene) there is evidence that DNA damage associated with metabolism can lead to 

16 mutations in cancer-related genes. Tumor promotion and progression by PAHs may involve 

17 parent compound binding to the aryl hydrocarbon (Ah) receptor and subsequent alterations of 

18 gene expression, as well as by cell proliferation in response to cytotoxic effects from metabolites 

19 (see Section 2.4, Similarities in Carcinogenic Mode of Action for PAHs). Thus, there is some 

20 evidence that an assumption of similar toxicological action is reasonable, but some aspects of the 

21 diversity of biological activities among PAHs are unexplained. The second assumption of no 

22 interactions at low levels of exposure may be reasonable, but some evidence of toxicological 

23 interactions among PAHs and among PAHs and other chemicals is available (see Section 2.8, 

24 Additivity of PAHs in Combined Exposures). 

25 Other key limitations to the RPF approach, relative to whole mixture approaches, are: 

26 (1) RPFs have been derived for a limited number of PAHs; and (2) cancer risks from non-PAH 

27 components, unidentified PAHs, and heterocyclic and substituted PAHs in PAH mixtures are not 

28 estimated. The first of these limitations is being addressed, to the degree allowable by available 

29 data, by the derivation of RPFs for numerous PAHs as discussed in Sections 4 and 5 of this 

30 report. If non-PAH carcinogenic components are identified and quantified in the complex 

31 mixture of concern and appropriate dose-response data are available, the second limitation can be 

32 addressed by using a response addition approach (i.e., adding the cancer risk from PAH 

33 components estimated by the RPF approach to cancer risks estimated for the non-PAH 

34 carcinogenic components of the mixture). Previous efforts to validate the RPF approach using 

35 data for PAH mixtures are discussed in Section 6.7. These validation efforts compared the 

36 cancer risk of a PAH mixture measured experimentally with the cancer risk that was predicted 

37 using the RPF method but were limited by the small number of compounds for which RPFs and 

 

1

Throughout this report, the term “tumorigenicity” is used to describe the production of either benign or malignant 

tumors. 
11 DRAFT – DO NOT CITE OR QUOTE

1 analytical data were available (Muller et al., 1997; McClure, 1996; Goldstein et al., 1994; 

2 Clement Associates, 1990, 1988; Krewski et al., 1989). Validation of the updated approach 

3 presented here wold be of value, either using previous data on PAH mixtures (human and 

4 animal) or using new data collected with the main purpose of evaluating the validity of the 

5 approach. 

6 

7 2.1. PAHs AS A CHEMICAL CLASS 

8 The PAH chemical class has been variously defined to include organic compounds 

9 containing either two or more, or three or more, fused rings made up of carbon and hydrogen 

10 atoms (i.e., unsubstituted parent PAHs and their alkyl-substituted derivatives) (WHO, 1998). 

11 Most PAHs are high-melting, high-boiling point, lipophilic compounds, predominately generated 

12 from the combustion or pyrolysis of organic matter. The PAH chemical class includes alkylated 

13 PAHs (e.g., 1,4-dimethylphenanthrene and 5-methylchrysene), but not heterocyclic compounds 

14 containing N, S, or O or PAHs substituted with N-, S-, or O-containing groups; these are 

15 included in a larger chemical class, often referred to as polycyclic aromatic compounds (PACs) 

16 (WHO, 1998). The number of chemicals that comprise the PAHs class is unknown; however, 

17 there are thought to be hundreds of individual PAHs present as components of complex mixtures 

18 (WHO, 1998). The analysis presented here is limited in focus to include only unsubstituted 

19 PAHs with three or more fused aromatic rings containing only carbon and hydrogen atoms, 

20 because these are the most widely studied members of the PAH chemical class. Naphthalene is a 

21 widely studied 2-ring PAH compound; however, a separate toxicological review and 

22 carcinogenicity assessment is being developed by the IRIS Program for this compound and it is 

23 not included in this RPF approach. The list of PAH compounds that were considered for 

24 inclusion in this analysis is presented in Table 2-1 along with the Chemical Abstracts Service 

25 Registry Numbers (CASRNs) and the abbreviations that are utilized in tables throughout the 

26 report.
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Benzo[a]pyrene 50-32-8 BaP 252.31 

Aceanthrylene 202-03-09 ACEA 202.26 

Acenaphthene 83-32-9 AN 154.21 

Acenaphthylene 208-96-8 ANL 152.20 

Acephenanthrylene 201-06-9 APA 202.26 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Acepyrene, 2,3- 25732-74-5 ACEP 228.29 

Anthanthrene 191-26-4 AA 276.34 

Anthracene 120-12-7 AC 178.23 

Benzacenaphthylene 76774-50-0 BAN 202.26 

Benz[a]anthracene 56-55-3 BaA 228.29 

Benzo[a]fluoranthene 203-33-8 BaF 252.32 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Benzo[a]fluorene 238-84-6 BaFE 216.28 

Benzo[a]perylene 191-85-5 BaPery 302.38 

Benz[b,c]aceanthrylene, 11H- 202-94-8 BbcAC 240.30 

Benz[b]anthracene 

(Naphthacene) 

92-24-0 BbA 228.29 

Benzo[b]chrysene 214-17-5 BbC 278.35 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Benzo[b]fluoranthene 205-99-2 BbF 252.32 

Benzo[b]fluorene, 11H 243-17-4 BbFE 216.28 

Benzo[b]perylene 197-70-6 BbPery 302.38 

Benzo[c]chrysene 194-69-4 BcC 278.35 

Benzo[c]fluorene 205-12-9 BcFE 216.28 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Benzo[c]phenanthrene 195-19-7 BcPH 228.29 

Benz[e]aceanthrylene 199-54-2 BeAC 252.32 

Benzo[e]pyrene 192-97-2 BeP 252.32 

Benzo[g,h,i]fluoranthene 203-12-3 BghiF 226.28 

Benzo[g,h,i]perylene 191-24-2 BghiP 276.34 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Benzo[g]chrysene 196-78-1 BgC 278.35 

Benz[j]aceanthrylene 202-33-5 BjAC 252.32 

Benzo[j]fluoranthene 205-82-3 BjF 252.32 

Benzo[k]fluoranthene 207-08-9 BkF 252.32 

Benz[l]aceanthrylene 211-91-6 BlAC 252.32 

Benzophenanthrene 65777-08-4 BPH 228.29 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Chrysene 218-01-9 CH 228.29 

Coronene 191-07-1 CO 300.36 

Cyclopent[h,i]aceanthrylene 131581-33-4 CPhiACEA 226.28 

Cyclopenta[c,d]pyrene 27208-37-3 CPcdP 226.28 

Cyclopenta[d,e,f]chrysene, 4H- 202-98-2 CPdefC 240.30 

Cyclopenta[d,e,f]phenanthrene 203-64-5 CPdefPH 190.24 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Cyclopenta[h,i]acephenanthrylene 114959-37-4 CPhiAPA 226.28 

Cyclopentaphenanthrene 219-08-9 CPPH 216.28 

Cyclopenteno-1,2-benzanthracene, 5,6- 7099-43-6 CPBA 268.36 

Dibenz[a,c]anthracene 

(Benzotriphenylene) 

215-58-7 DBacA 278.35 

Dibenzo[a,c]fluorene, 13H- 201-65-0 DBacFE 266.34 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Dibenzo[a,e]fluoranthene 5385-75-1 DBaeF 302.38 

Dibenzo[a,e]pyrene 192-65-4 DBaeP 302.38 

Dibenzo[a,f]fluoranthene 

(Indeno[1,2,3-fg]naphthacene

203-11-2 DBafF 302.38 

Dibenzo[a,g]fluorene, 13H- 207-83-0 DBagFE 266.34 

Dibenz[a,h]anthracene 53-70-3 DBahA 278.35 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Dibenzo[a,h]pyrene 189-64-0 DBahP 302.38 

Dibenzo[a,i]pyrene 189-55-9 DBaiP 302.38 

Dibenzo[a,l]pyrene 191-30-0 DBalP 302.38 

Dibenzo[b,e]fluoranthene 2997-45-7 DBbeF 302.38 

Dibenzo[e,l]pyrene 

(Dibenzo[fg,op]naphthacene)

192-51-8 DBelP 302.38 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Dibenzo[h,rst]pentaphene 192-47-2 DBhrstPent 352.43 

Dibenz[j,mno]acephenanthrylene 153043-82-4 DBjmnoAPH 276.34 

Dibenz[k,mno]acephenanthrylene 153043-81-3 DBkmnoAPH 276.34 

Dihydroaceanthrylene, 1,2- 641-48-5 DACEA 204.27 

Fluoranthene 206-44-0 FA 202.26 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Fluorene 86-73-7 FE 166.22 

Indeno [1,2,3-c,d] fluoranthene 193-43-1 IF 276.34 

Indeno[1,2,3-c,d]pyrene 193-39-5 IP 276.34 

Naphth[1,2,3-mno]acephenanthrylene 113779-16-1 N123mnoAPH 276.34 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Naphtho[1,2-b]fluoranthene 111189-32-3 N12bF 302.38 

Naphtho[2,1-a]fluoranthene 203-20-3 N21aF 302.38 

Naphtho[2,3-a]pyrene 

(Naphtho[2,1,8-qra]naphthacene) 

196-42-9 N23aP 302.38 

Naphtho[2,3-e]pyrene 

(Dibenzo[de,qr]naphthacene) 

193-09-9 N23eP 302.38 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Pentacene 135-48-8 PCE 278.35 

Pentaphene 

(Dibenzphenanthrene, 2,3:6,7-) 

222-93-5 Pent 278.35 

Perylene 198-55-0 Pery 252.32 

Phenanthrene 85-01-8 PH 178.23 

Picene 213-46-7 Pic 278.35 
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Table 2-1. PAHs evaluated in the RPF analysis 

PAH 

(common synonyms) CASRN Abbreviation Structure 

Molecular 

weight 

(g/mol) 

Pyrene 129-00-0 Pyr 202.26 

Tribenzofluoranthene 3,4-10, 11-12,13- 13579-05-0 TBF 352.43 

Triphenylene 217-59-4 Tphen 228.29 

1 

2 Unsubstituted PAHs have been further classified into alternant and nonalternant 

3 compounds. Alternant PAHs are those compounds composed solely of fused benzene rings, 

4 while nonalternant PAHs contain both benzene and five carbon rings. Among alternant PAHs, 

5 important structural features related to enhanced mutagenicity and carcinogenicity include the 

6 presence of at least four rings (Bostrom et al., 2002). Common structural features of PAH 

7 compounds are illustrated in Figure 2-1. 
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1 

2 Figure 2-1. Structural features of PAHs. 

3 

4 2.2. THE TOXICOLOGICAL DATABASE FOR PAHs 

5 Over the last 30- to 50-years, a large PAH database has been generated including studies 

6 of carcinogenicity in animal bioassays, genotoxicity in various test systems, and metabolism 

7 (bioactivation) to tumorigenic and/or genotoxic intermediates. Carcinogenicity and genotoxicity 

8 data are sufficient to classify a number of individual PAHs as possibly carcinogenic to humans 

9 (WHO, 1998; U.S. EPA, 1993; IARC, 1989, 1986, 1985, 1984a, b, 1983). Other PAHs have 

10 been tested for tumorigenicity and/or genotoxicity, but either negative or equivocal results were 

11 obtained; for many positive results were only observed in genotoxicity assays (e.g., pyrene). 

12 Many studies have been performed to provide further understanding about the carcinogenic 

13 mode of action of PAHs (see Bostrom et al., 2002; WHO, 1998; ATSDR, 1995). Therefore, the 

14 PAH database contains studies that evaluate: 

15 

C

H2

Benzo[a]pyrene Pyrene

Examples of Alternant PAHs

Examples of Nonalternant PAHs

Fluorene Fluoranthene

Bay-region and Fjord-regions of PAHs

Chrysene Benzo[c]phenanthrene

Bay-region Fjord-region
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1 • Metabolism to reactive intermediates; 

2 

3 • Characterization of PAH-DNA adducts; 

4 

5 • Mutagenicity of PAHs in bacterial and mammalian cells; 

6 

7 • Mutation spectra in identified oncogene and tumor suppressor genes; 

8 

9 • Clastogenic effects; 

10 

11 • Cell transformation; and 

12 

13 • Initiation and promotion of carcinogenicity. 

14 

15 A significant limitation to the database is the lack of data from long-term oral or 

16 inhalation cancer studies for most individual PAH compounds. In addition, benzo[a]pyrene is 

17 the only chemical for which long-term animal studies have been conducted by multiple exposure 

18 routes (Kroese et al., 2001; Culp et al., 1998, 1996a, b; Thyssen et al., 1981, 1980; Rigdon et al., 

19 1969; Rigdon and Neal, 1969, 1966; Neal and Rigdon, 1967). Furthermore, most of the 

20 toxicological data available for PAHs relate to cancer or genotoxicity. Available information on 

21 the systemic, noncarcinogenic effects of PAHs is limited although immunological, neurotoxic, 

22 and developmental effects have been noted in animal studies (for earlier reviews see WHO, 

23 1998; ATSDR, 1995). As a result, the relative potency methodology described here is applied 

24 only to cancer risk assessment for PAHs. 

25 

26 2.3. BENZO[A]PYRENE AS AN INDEX CHEMICAL 

27 Because long-term animal studies are not available for many individual PAHs, it is 

28 necessary to choose an appropriate index chemical for comparison of relative carcinogenic 

29 potency. The index compound is typically the best-studied member of the class, with the largest 

30 body of available data describing exposure and health effects. The index chemical should have a 

31 quantitative dose-response assessment of acceptable scientific quality and must have (or be 

32 expected to have) similar toxic effects to the rest of the members of the class. 

33 Although the PAH composition of complex mixtures varies, benzo[a]pyrene is 

34 considered to be present in significant amounts in certain occupational environments and urban 

35 settings (WHO, 1998; Petry et al., 1996; ATSDR, 1995). Benzo[a]pyrene is one of the most 

36 potent of the carcinogenic PAHs and has, therefore, been proposed to contribute significantly to 

37 the carcinogenicity of a PAH mixture, even when present in low concentrations (Petry, 1996). 

38 Benzo[a]pyrene is also the best-studied PAH compound, with carcinogenicity bioassay data 
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1 available for several routes of exposure and a considerable number of studies on carcinogenic 

2 mode of action. 

3 The laboratory animal database for benzo[a]pyrene is robust. Benzo[a]pyrene has been 

4 shown to induce tumors at the site of administration and at distal sites in numerous studies. 

5 Dose-response data for tumors are available for the oral, inhalation, and dermal routes of 

6 administration in multiple species. There are methodological limitiations associated with the 

7 inhalation data (Thyssen et al., 1981), although positive findings in intratracheal instillation 

8 studies support the observed positive response. Limited dermal exposure studies with several 

9 strains of mice also provide data on dose-related tumor incidences (Albert et al., 1991; 

10 Warshawsky and Barkley, 1987; Habs et al., 1984, 1980; Nesnow et al., 1983; Wynder et al., 

11 1957). 

12 The animal carcinogenicity database for benzo[a]pyrene includes several well-conducted 

13 oral cancer bioassays. Kroese et al. (2001) conducted a well-designed gavage study of 

14 benzo[a]pyrene carcinogenicity and found that benzo[a]pyrene induced tumors at multiple sites 

15 in rats of both sexes, specifically in the liver, forestomach, auditory canal, and oral cavity. In 

16 another well-conducted study, using Ah-responsive B6C3F1 female mice exposed in the diet 

17 (Beland and Culp, 1998; Culp et al., 1998), only portal-of-entry tumors were found, including 

18 papillomas and/or carcinomas of the forestomach, esophagus, tongue, and larynx. Earlier, Neal 

19 and Rigdon conducted a number of related studies evaluating the carcinogenicity of 

20 benzo[a]pyrene in feed in Ah-responsive white Swiss mice (Rigdon and Neal, 1969, 1966; Neal 

21 and Rigdon, 1967). These latter studies were not conducted using standard, modern 

22 toxicological methods and have limitations, including: inconsistent dosing protocols; varying 

23 ages of the animals; use of benzene as a solvent; small numbers of animals; and evaluation of 

24 only a limited number of tissues. However, the Neal and Rigdon studies provide useful dose25 response information on benzo[a]pyrene carcinogenicity. Following oral administration via 

26 feeding of benzo[a]pyrene, site-of-contact tumors, both papillomas and carcinomas, were 

27 induced in the forestomach, esophagus, and larynx of mice (Culp et al., 1998; Neal and Rigdon, 

28 1967) and rats (Brune et al., 1981). The results following inhalation, dermal, or oral exposure 

29 are further supported by numerous mechanistic studies or assays using infant mice, susceptible 

30 transgenic strains, or Ah-receptor knockout mice. 

31 Benzo[a]pyrene is a complete carcinogen and likely acts by initiating tumors through 

32 direct DNA damage as well as by promoting tumor growth. Benzo[a]pyrene has been shown to 

33 be mutagenic in multiple assay systems. Several modes of carcinogenic action are possible. 

34 These include: 

35 

36 (1) Alteration of pathways regulating cell proliferation and survival (Tannheimer et al., 

37 1998); 

38 
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1 (2) Inhibition of intracellular communication (Sharovskaia et al., 2003; Blaha et al., 2002; 

2 Rummel et al., 1999); 

3 

(3) Altered intracellular Ca2+ 4 signaling (Tannheimer et al., 1998); 

5 

6 (4) Modulation of cell survival, cell proliferation, and altered growth via generation of 

7 oxidative stress and activation of oxidant stress signaling (Burdick et al., 2003; Miller 

8 and Ramos, 2001); 

9 

10 (5) Altered apoptosis processes (Chen et al., 2003); 

11 

12 (6) Dysregulation of normal circulating hormone levels or activity affecting tumorigenesis in 

13 reproductive tissues (Safe and Wormke, 2003; Archibong et al., 2002) or the central 

14 nervous system (Dasgupta and Lahiri, 1992); 

15 

16 (7) Disruption of cell cycle kinetics in breast cancer cells (Jeffy et al., 2002, 2000); and 

17 

18 (8) Disruption of DNA repair through alteration of RNA polymerase activity (Shah and 

19 Bhattacharya, 1989). 

20 

21 Oral (dietary) carcinogenicity bioassays are available that compare manufactured gas 

22 plant (MGP) residue (Weyand et al., 1995) or coal tar preparations (Culp et al., 1998; Gaylor et 

23 al., 1998) with benzo[a]pyrene. In both cases, there were significant differences in target organ 

24 distribution of tumors between benzo[a]pyrene and complex mixtures of PAHs. Following 

25 dietary administration, benzo[a]pyrene-induced tumors were observed primarily at the point of 

26 contact (i.e., the forestomach), while MGP residue and coal tar produced tumors in the lung, 

27 liver, forestomach, skin, and other organs. Tissue-specific differences in metabolic activation 

28 and DNA binding of PAHs may contribute to the observed differences in target organ sensitivity 

29 (Weyand and Wu, 1995; Culp and Beland, 1994). A recent gavage study in rats (Kroese et al., 

30 2001) demonstrated that oral exposure to benzo[a]pyrene could induce tumors at distal sites (i.e., 

31 liver, auditory canal); however, no lung tumors were observed. The lung appears to be a 

32 sensitive target organ for complex mixture carcinogenicity, but is insensitive to benzo[a]pyrene33 induced tumorigenicity via oral and dermal exposures. The existing data limitations for other 

34 PAHs, however, necessitate the use of benzo[a]pyrene as the only appropriate index chemical for 

35 PAHs. 

36 In summary, benzo[a]pyrene is the most appropriate compound to use as an index 

37 chemical for carcinogenic PAHs. It is well-studied, with a robust database of both bioassay data 

38 and mode of action information. Benzo[a]pyrene is a complete carcinogen with both initiating 

39 and promoting properties, is among the most potent PAH carcinogens, and is prevalent in many 

40 complex environmental mixtures. No alternative index chemical was identified from the list of 

41 target PAHs. 

42 
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1 2.4. SIMILARITIES IN MODE OF CARCINOGENIC ACTION FOR PAHs 

2 A common mode of action for chemicals is the basis for the assumption of dose additivity 

3 that underlies the RPF approach (U.S. EPA, 1990). The carcinogenic mode of action for PAHs 

4 has been extensively reviewed (Ramesh, 2004; CCME, 2003; Bostrom et al., 2002; Larsen and 

5 Larsen, 1998; WHO, 1998; Muller et al., 1997; Sjogren et al., 1996; ATSDR, 1995; Malcolm 

6 and Dobson, 1994; U.S. EPA, 1990). The major key events that have been associated with PAH 

7 carcinogenicity include: 

8 

9 • Oxidative metabolism to reactive intermediates that covalently bind to DNA, RNA, and 

10 proteins (benzo[a]pyrene metabolism is illustrated in Figure 2-2); 

11 

12 • Formation of DNA adducts; 

13 

14 • Tumor initiation due to mutations in cancer-related genes (e.g., tumor suppressor genes 

15 or oncogenes); and 

16 

17 • Tumor promotion related to cytotoxicity and formation of reactive oxygen species, and 

18 Ah receptor (AhR) affinity and upregulation of genes related to biotransformation, 

19 growth, and differentiation. 

20 
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1 

2 

3 

4 Source: Miller and Ramos (2001). 

5 

6 Figure 2-2. Metabolic pathways for benzo[a]pyrene.
33 DRAFT – DO NOT CITE OR QUOTE

1 Formation of reactive intermediates and DNA adducts

2 Each of the key events identified above is affected by the chemical structure of the 

3 individual PAH. At least three distinct molecular mechanisms have been proposed to explain the 

4 tumor initiation process of PAHs (Xu et al., 2009; Jiang et al., 2007, 2005; Xue and 

5 Warshawsky, 2005; Bolton et al., 2000; Penning et al., 1999; Harvey, 1996; Cavalieri and 

6 Rogan, 1995). These modes of action include the formation of diol epoxides, radical cations, 

7 and o-quinones (Figure 2-3). Diol epoxide formation leads to stable and unstable DNA adducts, 

8 mainly at guanine and adenine, which can lead to mutations in proto-oncogenes and tumor9 suppressor genes. Radical cation formation may lead to the generation of unstable adducts at 

10 guanine and adenine, leading to apurinic sites and mutation in HRAS. Orthoquinone formation 

11 could lead to stable and unstable DNA adducts and generation of reactive oxygen species, 

12 inducing mutations in P53. The evidence supporting the role of these reactive metabolites in 

13 tumor initiation includes a characterization of the specific DNA adducts arising from PAH 

14 metabolism and observations of mutagenesis resulting from direct exposure. Figure 2-3 

15 illustrates the proposed key steps in the mode of action for PAH carcinogenesis. These include 

16 the interaction of reactive metabolites with DNA to form adducts, induction of depurination, 

17 transversion mutations (e.g., GC→TA or AT→TA), and oxidative damage to DNA, and tumor 

18 promotion mediated by AhR-mediated effects on gene regulation. 
34 DRAFT – DO NOT CITE OR QUOTE

1 

2 

3 

4 Figure 2-3. Overview of the proposed key events in the mode of action for 

5 PAH carcinogenicity. 

6 

7 The formation of diol epoxides is a proposed key step in the most established mode of 

8 action for PAH-induced carcinogenicity. Extensive studies of the metabolism of carcinogenic 

9 PAHs suggest that bay-region and fjord-region diol epoxides are some of the ultimate reactive 

10 metabolites of PAHs (Jerina et al., 1978; Jerina and Lehr, 1977). These metabolites are 

11 generally formed through cytochrome P450 (CYP) oxidation to form epoxides and epoxide 

12 hydrolase cleavage resulting in diol formation. CYP1A1 appears to be the primary isozyme 

13 involved in diol epoxide formation; however, other isozymes may also contribute to PAH 

14 metabolism (i.e., CYPIA2, CYP1B1, CYP3A4) (Bostrom et al., 2002; ATSDR, 1995). Non15 alternant PAHs, composed of fused benzenoid and five-membered rings, may be metabolized 

16 through other pathways resulting in the formation of reactive intermediates that bind to DNA. 

17 Classic bay- and fjord-region diol epoxides may be formed from these compounds; however, 

18 epoxide formation at cyclopenta-ring structures has also been demonstrated to result in DNA 

19 adduct formation (Bostrom et al., 2002). 

20 Many studies have been performed to evaluate the formation of DNA adducts following 

21 in vivo or in vitro exposure to PAHs. Diol epoxide metabolites interact preferentially with the 

PAH

diol epoxide

radical cation

o-quinone and 

ROS

DNA Adducts

DNA Adducts

DNA Adducts 

and oxidative 

base damage

Mutation 

(transversion )

Mutation 

(depurination)

Mutation 

(depurination , 

oxidative damage, 

and strand 

scission)

Proliferation of 

initiated cells Neoplasm

Inflammatory

response

Cytotoxicity 

OH

OH

O

+

Exposure Metabolism

O

O

Initiation Promotion Progression

Key Events in the Mode of Action for PAH Carcinogenicity

Binding to Ah

Receptor

Upregulation of 

genes related to 

biotransformation,

growth, and 

differentiation
35 DRAFT – DO NOT CITE OR QUOTE

1 exocyclic amino groups of deoxyguanine and deoxyadenine (Geacintov et al., 1997; Jerina et al., 

2 1991). Adducts may give rise to mutations, unless these adducts are removed by DNA repair 

3 processes prior to replication. The stereochemical nature of the diol epoxide metabolite (i.e., 

4 anti- versus syn-diol epoxides) affects the number and type of adducts and mutation that occurs. 

5 Figure 2-4 presents the structures of four stereoisomeric adducts arising from the interaction of 

6 benzo[a]pyrene diol epoxide metabolites with the deoxyguanosine (dG) residues in DNA 

7 (Geacintov et al., 1997). Transversion mutations (e.g., GC→TA or AT→TA) are the most 

8 common type of mutation found in mammalian cells following diol epoxide exposure (Bostrom 

9 et al., 2002). 

10 

11 Source: Geacintov et al. (1997). 

12 

13 Figure 2-4. Structures of the four stereoisomeric adduct moieties, 

anti-[BaP]-N

2

14 -dG, derived from the trans- or cis-covalent binding of 

15 (+)-anti-BaP diol epoxide or (-)-anti-BaP diol epoxide to dG residues in DNA. 

16 

17 Radical cation formation involves a one-electron oxidation that produces electrophilic 

18 radical cation intermediates (Cavalieri and Rogan, 1995, 1992). Oxidation of this type can occur 

19 by CYP or peroxidase enzymes (i.e., horseradish peroxidase, prostaglandin H synthetase). 

HO

HO

OH

NH

NH

N

N

N

O

R HO

HO

OH

NH

NH

N

N

N

O

R

HO

HO

OH

NH

NH

N

N

N

O

R

HO

HO

OH

NH

NH

N

N

N

O

R

10S

β'

α'

10R

10R

10S

10R (+)-cis-anti-[BaP]-N2-dG 10S (-)-cis-anti-[BaP]-N2-dG

10S (+)-trans-anti-[BaP]-N2-dG 10R (-)-trans-anti-[BaP]-N2-dG
36 DRAFT – DO NOT CITE OR QUOTE

1 Radical cations can be further metabolized to phenols and quinones (Cavalieri et al., 1988a), or 

2 they can form unstable adducts with DNA that ultimately result in depurination (Cavalieri et al., 

3 2005, 1993; Rogan et al., 1993). Radical cations have been shown to play a major role in 

4 formation of DNA adducts for several carcinogenic PAHs (e.g., 7,12-dimethylbenzanthracene, 

5 benzo[a]pyrene, dibenzo[a,l]pyrene). The predominant depurinating adducts occur at the N-3 

6 and N-7 positions of adenine and the C-8 and N-7 positions of guanine (Cavalieri and Rogan, 

7 1995; Li et al., 1995). Figure 2-5 illustrates three depurinating adducts of benzo[a]pyrene 

8 formed by one-electron oxidation. Abasic sites resulting from base depurination undergo error9 prone excision repair to induce mutations. In the case of DBalP-treated mouse skin, repair error 

10 from abasic sites resulted in H-ras oncogene mutations that underwent rapid clonal expansion 

11 and regression (Chakravarti et al., 2000). H-ras mutations in mouse skin papillomas also 

12 corresponded to adenine and guanine depurinating adducts resulting from exposure to DBalP, 

13 7,12-dimethyl-benz[a]anthracene, benzo[a]pyrene, and benzo[a]pyrene-7,8-dihydrodiol 

14 (Chakravarti et al., 2008). 

15 

16 

17 Source. Cavalieri and Rogan (1995). 

18 

19 Figure 2-5. Depurinating adducts of benzo[a]pyrene formed by one-electron 

20 oxidation. 

21 

22 o-Quinone metabolites of PAHs are formed by enzymatic dehydrogenation of 

23 dihydrodiols (Bolton et al., 2000; Penning et al., 1999; Harvey, 1996; ATSDR, 1995). 

24 Dihydrodiol dehydrogenase enzymes are members of the α-keto reductase gene superfamily. 

25 o-Quinone metabolites are potent cytotoxins, are weakly mutagenic, and are capable of 

26 producing a broad spectrum of DNA damage. These metabolites can interact directly with DNA 

27 and also result in production of reactive oxygen species (i.e., hydroxyl and superoxide radicals) 

28 that may produce further cytotoxicity and DNA damage. The DNA damage caused by 

29 o-quinones may include the formation of stable adducts (Balu et al., 2006), N-7 depurinating 

30 adducts (McCoull et al., 1999), oxidative base damage (i.e., 8-oxo-2’-dG or 8-oxo-dG) (Park et 

HN

N N

H

N

H2N

O

HN

N N

N

H2N

O

N

N N

N

NH2

BaP-6-C8-guanine BaP-6-N7-guanine BaP-6-N7-adenine
37 DRAFT – DO NOT CITE OR QUOTE

1 al., 2006, 2005), and strand scission (Flowers-Geary et al., 1997). The reactive oxygen species 

2 generated by the o-quinone of benzo[a]pyrene and other PAH o-quinones have been shown to 

3 induce mutation in the p53 tumor suppressor gene (Park et al., 2008; Shen et al., 2006; Yu et al., 

4 2002). Figure 2-6 illustrates the spectrum of DNA adducts associated with PAH o-quinones. 

5 

6 

7 Source: Bolton et al. (2000). 

8 

9 Figure 2-6. Spectrum of DNA adducts anticipated with PAH o-quinones. 

10 

11 The cytotoxicity of o-quinone metabolites may also contribute to tumor promotion via 

12 inflammatory responses leading to cell proliferation (Burdick et al., 2003). 

13 

14 Genotoxicity and mutagenicity 

15 The genotoxicity and mutagenicity of PAHs have been demonstrated in various bacterial 

16 and mammalian assays (see Section 4.3.2 below) (reviewed in WHO, 1998; ATSDR, 1995). 

17 Mutagenesis of PAHs in the Ames assay (Salmonella typhimurium) as well as other bacterial 

HN

N

N

N

O

OPO3R

H2N

O

R3OPO

HN

N

N

N

O

OPO3R

H2N

O

R3OPO

O

O O

O

O

HN

N

N

N

O

H2N

OPO3R

RO3PO

OH

O

HN

N

N

N

O

H2N

OPO3R

RO3PO

HO

O B

O

OH OPO3R

RO3PO

B

O O

HN

N

N

N

O

H2N

OPO3R

OH

RO3PO

HN

N

N

N

O

H2N

O

O

NH

O

O

HN

N N

N

O

O

o-Quinone adducts ROS modifications

H

8'-oxo-dG base propenals

base pair mismatch 

G to T transversions

depurinating adducts

apurinic sites

G to T transversions

stable adducts

translesional synthesis

G to T transversions

RO3PO

RO3PO
38 DRAFT – DO NOT CITE OR QUOTE

1 assays requires the presence of a mammalian metabolic enzyme system. In most cases, this is 

2 supplied by postmitochondrial supernatant (S9) from the liver of rodents treated with an enzyme 

3 inducer. Mammalian cell mutagenesis in Chinese hamster V79 cells and mouse lymphoma 

4 L5178Y cells also requires metabolic activation in the form of a rodent S9 mix or co-cultivation 

5 with metabolically active rodent cells (i.e., cell-mediated assay). Several studies have noted a 

6 correlation between mutagenic potency and tumor initiation potency in the 2-stage dermal 

7 carcinogenicity assay for multiple PAH compounds (LaVoie et al., 1985, 1979; Raveh et al., 

8 1982). 

9 

10 Tumor promotion and the AhR 

11 The ability of certain PAHs to act as tumor promoters as well as initiators may increase 

12 their carcinogenic potency (Andrews et al., 1978). The promotional effects of PAHs appear to 

13 be related to AhR affinity and the upregulation of genes related to biotransformation (i.e., 

14 induction of CYP1A1), growth, and differentiation (Bostrom et al., 2002). Figure 2-7 illustrates 

15 the function of the AhR and depicts the genes regulated by this receptor as belonging to two 

16 major functional groups (i.e., induction of metabolism or regulation cell differentiation and 

17 proliferation). PAHs bind to the cytosolic AhR in complex with heat shock protein 90 (Hsp90). 

18 The ligand-bound receptor is then transported to nucleus in complex with the AhR nuclear 

19 translocator protein (ARNT). The AhR complex interacts with the Ah responsive elements of 

20 the DNA (AhREDNA) to increase the transcription of proteins associated with induction of 

21 metabolism and regulation of cell differentiation and proliferation. 

22 
39 DRAFT – DO NOT CITE OR QUOTE

AHR Hsp90 AHR

Hsp90

AHR Hsp90

Hsp90

ARNT

Hsp90 ARNT

Hsp90

AHR

ARNT

AHREDNA

PAH

nucleus

Enhanced

specific

mRNA

production

Increased

synthesis of 

PAH metabolizing 

enzymes

Increased 

synthesis of 

proteins that

regulate cell

differentiation and

proliferation

1 

2 AhRAhRSource: Okey et al. (1994). 

3 

4 Figure 2-7. Interaction of PAHs with the Ah receptor – regulation of genes 

5 related to induction of metabolism and cell differentiation and proliferation. 

6 

7 In general, it has been demonstrated that fjord-region PAHs are strong mutagens and 

8 carcinogens, but have a low binding affinity to the AhR. Conversely bay-region PAHs possess a 

9 greater affinity for AhR binding, and are better tumor promoters in carcinogenicity bioassays 

10 (Bostrom et al., 2002). CYP1A1 induction by PAHs is considered to contribute to tumorigenesis 

11 by increasing the production of DNA-reactive metabolites (Ayrton et al., 1990). However, 

12 several recent studies indicate that CYP1A1 induction potency does not correlate well with 

13 carcinogenic potency. These studies compared CYP1A1 induction potency for several PAHs 

14 using assays to measure ethoxyresorufin O-deethylase (EROD) activity, CYP1A1 protein, and 

15 mRNA levels, or chemical-activated luciferase reporter gene expression (Bosveld et al., 2002; 

16 Machala et al., 2001; Bols et al., 1999; Till et al., 1999; Willett et al., 1997). 

17 

18 Tumor promotion and cytotoxicity

19 PAHs are metabolized to o-quinones, which are cytotoxic and can generate reactive 

20 oxygen species (Bolton et al., 2000; Penning, 1999). PAH o-quinones reduce the viability and 

21 survival of rat and human hepatoma cells (Flowers-Geary et al., 1996, 1993). Inflammatory 
40 DRAFT – DO NOT CITE OR QUOTE

1 responses to cytotoxicity may contribute to the tumor promotion process. For example, 

2 benzo[a]pyrene quinones (1,6-, 3,6-, and 6,12-benzo[a]pyrene-quinone) generated reactive 

3 oxygen species and increased cell proliferation by enhancing the epidermal growth factor 

4 receptor pathway in cultured breast epithelial cells (Burdick et al., 2003). Dermal exposure of 

5 mice to DBalP and dimethyl-benz[a]anthracene resulted in an inflammatory response that was 

6 correlated with epidermal hyperplasia and skin tumor promotion (Casale et al., 2000, 1997). The 

7 extent of epidermal hyperplasia was correlated with the cytokine mRNA response in lymph 

8 nodes and skin of treated mice (Casale et al., 2000). 

9 

10 Genetic targets and tumor formation 

11 DNA adducts and oncogenes/tumor suppressor gene mutations have been demonstrated 

12 in tumor tissue from humans and laboratory animals. DeMarini et al. (2001) demonstrated 

13 mutations in the p53 tumor suppressor gene and the K-ras oncogene in the lung tumors of 

14 nonsmokers, whose tumors were associated with exposure to smoky coal. Lung tumors were 

15 obtained from 24 nonsmoking women from China (age 30–63, mean age 48.5 + 8.8 years) who 

16 used smoky coal in their homes without chimneys. Bronchioloalveolar adenocarcinoma and 

17 acinar adenocarcinoma were observed in 54 and 46% of the women studied, respectively. The 

18 observed mutations in lung tumors were primarily G→T transversions at either K-ras or p53. 

19 Mutation hotspots in the lung tumors examined corresponded with hot spots for PAH adducts 

20 (codon 154), cigarette smoke associated mutations (codon 249), and both of these events 

21 together (codon 273). The mutation spectrum was described as unique and consistent with 

22 exposure to PAHs in the absence of cigarette smoke.

23 Mutations in the K-ras, H-ras, and p53 genes were assessed in forestomach tumors 

24 (n = 31) of mice fed benzo[a]pyrene in the diet (0, 5, 25, or 100 ppm) for 2 years (Culp et al., 

25 2000). Forestomach tumors had K-ras mutations (68% of tumors), which were G→T or 

26 C transversions in codon 12 or 13. H-ras (codon 13) and p53 mutations characterized as G→T 

or C transversions were also each found in 10% of forestomach tumors. [32 27 P]-postlabeling of 

28 forestomach DNA of benzo[a]pyrene-treated mice revealed one major adduct characterized as 

dG-N229 -BPDE. There was a linear relationship between the amount of benzo[a]pyrene consumed 

and the concentration of dG-N230 -BPDE in the forestomach of mice. For benzo[a]pyrene, 

31 forestomach tumor incidence increased sharply with adduct concentrations between 50 and 

32 140 fmol/mg DNA and in coal-tar fed mice. Tumor incidence increased sharply with 

dG-N233 -BPDE adduct levels between 20 and 60 fmol/mg DNA. The same levels of adduct were 

34 present in lung and liver of benzo[a]pyrene-treated mice, although only the forestomach 

35 exhibited benzo[a]pyrene-induced tumors (Goldstein et al., 1998). The presence of adducts in 

36 tumor-free tissue suggests that DNA adduct levels alone are not necessarily predictors of tumor 

37 outcome. 
41 DRAFT – DO NOT CITE OR QUOTE

1 A series of experiments designed to evaluate the mechanistic relationship between PAH 

2 DNA adducts, oncogene mutations, and lung tumorigenesis were performed in the A/J mouse 

3 lung model (Nesnow et al., 1998a, 1996, 1995; Mass et al., 1993). Tumorigenic potency in the 

4 lung of A/J mice varied over 2 orders of magnitude following a single intraperitoneal injection of 

5 seven PAHs of varying structure (benzo[a]pyrene, benzo[b]fluoranthene, benz[j]aceanthrylene, 

6 dibenz[a,h]anthracene, dibenzo[a,l]pyrene, cyclopenta[c,d]pyrene, and 5-methylchrysene). 

7 When considering the non-alkylated PAHs, the number of lung adenomas per mouse was highest 

8 for benz[j]aceanthrylene and cyclopenta[c,d]pyrene, each of which contain a pentacyclic ring 

9 feature. The major DNA adducts identified in the mouse lung included: 

10 

11 (1) Bay region diol epoxide adducts for benzo[a]pyrene, dibenz[a,h]anthracene, and 

12 5-methylcholanthrene; 

13 

14 (2) Phenolic diol epoxide adducts for benzo[b]fluoranthene; 

15 

16 (3) Cyclopenta-ring adducts for cyclopenta[c,d]pyrene and benz[j]aceanthrylene; 

17 

18 (4) Bisdihydrodiol epoxide adducts for dibenz[a,h]anthracene; and 

19 

20 (5) Fjord-region diol epoxide adducts for dibenzo[a,l]pyrene (Nesnow et al., 1998a, 1996, 

21 1995; Mass et al., 1993). 

22 

23 Guanine adducts were most common for all PAHs; however, adenine adducts were also 

24 demonstrated for dibenzo[a,l]pyrene and benz[j]aceanthrylene. Quantitative analysis of DNA 

adducts by [32 25 P]-postlabeling illustrates the importance of measuring DNA adduct levels over 

26 time. A time-integrated DNA adduct level (TIDAL) was linearly related to the dose of a 

27 particular PAH. The relationship of TIDAL level to tumor formation was similar for PAHs that 

28 produce different types of adducts and different mutations in the Ki-ras oncogene. This suggests 

29 that the probability of tumor formation for these PAHs may be related to the extent of overall 

30 DNA damage and repair rather than the formation of specific adduct at specific sites. 

31 The DNA sequence analysis of Ki-ras mutations in lung adenomas at codons 12 and 61 

32 was generally consistent with the DNA adduct data in that PAHs that produced guanine adducts 

33 also produced Ki-ras guanine mutations (Nesnow et al., 1998a, 1996, 1995; Mass et al., 1993). 

34 Cyclopenta[c,d]pyrene, benz[j]aceanthrylene, and 5-methylchrysene produced large numbers of 

35 adenomas per mouse (>90) and also produced a large proportion of tumors with CGT mutations 

36 at Ki-ras codon 12. Cyclopenta-ring adduct formation by cyclopenta[c,d]pyrene and 

37 benz[j]aceanthrylene was correlated with the formation of GGT→CGT mutations at Ki-ras 

38 codon 12. The primary mutation type for benzo[a]pyrene, benzo[b]fluoranthene, and 

39 dibenzo[a,l]pyrene was the GGT→TGT mutation, which is associated with the formation of diol 

40 epoxide guanine adducts. Dibenz[a,h]anthracene did not induce mutations in Ki-ras codons 12 
42 DRAFT – DO NOT CITE OR QUOTE

1 or 61; however, diol epoxide guanine adducts and lung adenomas in A/J mice were observed. 

2 This suggests that a different genetic target may be involved in carcinogenicity of this 

3 compound. 

4 H-ras mutations were studied in skin papillomas of SENCAR mice resulting from dermal 

5 initiation by benzo[a]pyrene or benzo[a]pyrene-7,8-dihydrodiol (400 nmol) followed by 12-O6 tetra-decanoylphorbol-acetate (TPA) promotion (Chakravarti et al., 2008). PCR amplification of 

7 the H-ras gene and sequencing revealed that codon 13 (GGC to GTC) and codon 61 (CAA to 

8 CTA) mutations in papillomas corresponded to the relative levels of depurinating adducts of 

9 guanine and adenine, despite the formation of significant amounts of stable DNA adducts. 

10 Other studies also suggest that multiple genetic targets may be involved in PAH 

11 mutagenicity and carcinogenicity (Conney et al., 2001; Smith et al., 2000). Smith et al. (2000) 

12 indicated that diol epoxide adducts and mutations were observed in the p53 tumor suppressor 

13 gene following in vitro exposure of cultured human bronchial epithelial cells to metabolites of 

14 benzo[a]pyrene, chrysene, benzo[c]phenanthrene, and benzo[g]chrysene. PAH adducts and 

15 corresponding mutations preferentially formed at lung mutational hot spots (codons 154, 157, 

16 158, 245, 248, and 273), suggesting that PAHs may contribute to the mutation spectrum 

17 observed in human lung cancer. Conney et al. (2001) provided evidence that dose-dependent 

18 differences may exist for the mutation spectra seen in PAH-induced tumors. Skin papillomas 

19 induced by benzo[a]pyrene in female mice were examined for mutations in the c-Ha-ras proto20 oncogene. The major difference between high- and low-dose groups was mutations at exon 2 of 

21 the c-Ha-ras gene, with the proportion of AT base pair mutations higher in the low-dose group. 

22 Dose-dependent changes in mutation profile were also evident in Chinese hamster V79 cells 

23 exposed to the diol epoxides of benzo[a]pyrene and benzo[c]phenanthrene (i.e., the proportion of 

24 AT mutations decreased with increasing concentration). 

25 In conclusion, the available data indicate that there may be multiple mechanisms of 

26 PAH-induced carcinogenicity. However, a common mode of action involving oxidative 

27 metabolism to reactive intermediates, DNA adduct formation, and subsequent mutagenic events 

28 is considered to be the primary mode of carcinogenic action. For these reasons, the use of a RPF 

29 approach to estimate cancer risk associated with PAH exposure is considered appropriate. The 

30 uncertainties and limitations related to the mode of action assumption for PAH-induced cancer 

31 are further discussed in Section 8.5. 

32 

33 2.5. STRUCTURAL ALERTS FOR PAH CARCINOGENESIS 

34 The carcinogenic activity of PAH compounds is influenced by specific structural 

35 features. For example, alternant PAHs having four or more benzene rings exhibit greater 

36 carcinogenic potency than PAHs with two or three benzene rings (Bostrom et al., 2002). The 

37 carcinogenic activity of PAHs is also related to the specific arrangement of the benzene rings. 

38 As described in Section 2.4, PAHs that form bay- and fjord-region diol or dihydrodiol epoxides 
43 DRAFT – DO NOT CITE OR QUOTE

1 are more potent carcinogens compared with linear PAHs that lack this structural feature 

2 (Bostrum et al., 2002). These metabolites are resistant to detoxification due to stereochemical 

3 effects and, consequently, are more likely to be mutagenic and cause cancer (Flesher et al., 1976; 

4 Buterin et al., 2000; Chang et al., 1981; Buening et al., 1979; MacLeod et al., 1979). 

5 Dihydrodiol epoxides formed at other positions on the PAH molecule (i.e., not the bay or fjord6 regions) are more accessible to glutathione transferase detoxification and are less potent 

7 mutagens and carcinogens (Flesher et al., 1976; MacLeod et al., 1979). Nonalternant PAHs 

8 containing fused benzenoid and five-membered rings, can also be metabolized to bay- and fjord9 region diol epoxides (Bostrum et al., 2002); however, epoxide formation at the cyclopenta-ring 

10 structure may also contribute to carcinogenicity (Bostrum et al., 2002; Nyholm et al., 1996). 

11 PAHs with at least four rings and a classic bay- or fjord-region (formed entirely by 

12 benzene rings; see Figure 2-1) may be characterized as containing structural alerts for 

13 carcinogenesis. However, this structural characterization is likely to be overly simplistic and 

14 other features may be important to carcinogenesis. Recent studies have applied quantitative 

15 structure activity relationship (QSAR) methods to evaluate the relationship between specific 

16 PAH structural features and mechanistic events related to carcinogenesis (Bruce et al., 2008; 

17 Vijayalakshmi et al., 2008). 

18 

19 2.6. SIMILARITIES IN RELATIVE POTENCY ACROSS ENDPOINTS 

20 Studies that have evaluated the association between cancer-related endpoints and 

21 tumorigencity of PAHs are briefly summarized below. 

22 Several studies have been performed that compare the bacterial or mammalian cell 

23 mutagenicity of various PAHs with tumor initiating activity or complete carcinogenesis (LaVoie 

24 et al., 1985, 1981, 1979; Raveh et al., 1982; Andrews et al., 1978). In general, mutagenicity 

25 appears to correlate best with tumor initiation. Complete carcinogenicity is not well-predicted by 

26 positive findings in short-term mutagenicity assays. Andrews et al. (1978) tested 24 PAHs for 

27 bacterial mutagenicity in the Ames test and compared these findings to evidence of 

28 carcinogenicity (parent and metabolites) from previously published studies. Positive findings of 

29 both mutagenicity and carcinogenicity were only reported for 14 of the 24 PAHs evaluated. 

30 Eight of the 10 remaining PAHs were found to be mutagenic in the Ames assay, but were not 

31 carcinogenic in animal studies. LaVoie et al. (1979) compared the mutagenicity, tumor-initiating 

32 activity, and complete carcinogenicity of several series of structurally related PAHs. Tumor33 initiating activity was found to correspond with complete carcinogenicity. Quantitation of 

34 mutagenicity in the Ames assay for structurally related PAHs failed to provide a reliable 

35 indication of tumor-initiating activity or complete carcinogenicity. In addition, mutagenicity 

36 results could not be used to predict which PAHs would be noncarcinogenic. Many PAHs were 

37 active mutagens, but were not shown to be carcinogenic. Studies using methylated derivatives of 

38 anthracene demonstrated a correlation between mutagenicity of specific metabolites and tumor 
44 DRAFT – DO NOT CITE OR QUOTE

1 initiating activity in mouse skin (LaVoie et al., 1985). Raveh et al. (1982) reported that the 

2 mutagenic response to PAHs in Chinese hamster V79 cells was similar to the skin tumor 

3 initiating activity observed in SENCAR mice. Benzo[a]pyrene was demonstrated to be a more 

4 potent mutagen and skin tumor initiator than cyclopenta[c,d]pyrene. 

5 Sjogren et al. (1996) performed a multivariate analysis to evaluate the relevance of 

6 different biological assays to the tumor initiating and promoting properties of PAHs. This 

7 analysis considered carcinogenicity (strength of evidence), bacterial mutagenicity, inhibition or 

8 enhancement of bacterial mutagenicity, AhR affinity, and enzyme induction. A principle 

9 components analysis indicated that bacterial mutagenicity data were poorly correlated with 

10 cancer bioassay data (regression coefficients ranged from -0.1 to 0.15). Variables describing 

11 AhR affinity showed the highest correlation with cancer data. A partial least squares regression 

12 analysis showed that all of the AhR affinity variables analyzed in the report were statistically 

13 relevant to describe cancer potency (regression coefficients ranged from 0.1 to 0.25). The 

14 structural requirements for AhR affinity are the same as those required for enzyme induction and 

15 bioactivation of PAHs. This analysis suggests that different chemical species (i.e., parent 

16 compounds or metabolites) may be responsible for the initiating and promoting properties of 

17 PAHs. Sjogren et al. (1996) proposed that mutagenicity reflects the cancer initiation potency, 

18 which may be more relevant at lower environmental exposure levels, and AhR affinity reflects 

19 the promoting effect of some PAHs that occur primarily at high doses in animal bioassays. 

20 Bostrom et al. (2002) suggested that the ability for a PAH to act as a promoter strongly increases 

21 its carcinogenic potency in animal studies. However, highly mutagenic fjord-region PAHs are 

22 potent carcinogens, despite a lower AhR affinity (Bostrom et al., 2002; Jerina et al., 1991). 

23 CYP1A1 induction by PAHs is considered to contribute to tumorigenesis by increasing 

24 the production of DNA-reactive metabolites (Ayrton et al., 1990). However, CYP1A1 induction 

25 potency alone does not appear to correlate well with carcinogenic potency of PAHs. EROD 

26 activity was evaluated as a measure of CYP1A1 induction in rat hepatocytes (Bosveld et al., 

27 2002; Till et al., 1999; Willett et al., 1997) and trout liver cells (Bols et al., 1999). Till et al.

28 (1999) additionally measured levels of CYP1A1 protein and mRNA. Machala et al. (2001) 

29 measured PAH activation of the AhR using a chemical-activated luciferase reporter gene assay. 

30 Comparable results were observed across studies and benzo[k]fluoranthene was consistently 

31 demonstrated to be the most potent inducer of CYP1A1. Chrysene, benzo[b]fluoranthene, 

32 dibenz[a,h]anthracene, and indeno[1,2,3-c,d]pyrene were also demonstrated to be more potent 

33 inducers of CYP1A1 than benzo[a]pyrene. However, most of these PAH compounds (except 

34 dibenz[a,h]anthracene) are considerably less potent as carcinogens in animal bioassays. 

35 Ross et al. (1995) evaluated the relationship between TIDAL values for DNA adduct 

36 formation and lung adenoma formation in A/J mice. The TIDAL value vs. tumor relationship 

37 was similar for five different PAHs, suggesting a correlation between adduct levels and tumor 

38 formation (regression analysis was not performed). As described above, the relationship of 
45 DRAFT – DO NOT CITE OR QUOTE

1 TIDAL level to tumor formation was similar for PAHs that produce different types of adducts 

2 and different mutations in the Ki-ras oncogene, suggesting that the probability of tumor 

3 formation may be related to the extent of overall DNA damage and repair (Nesnow et al., 1998a, 

4 1996, 1995; Mass et al., 1993). 

5 To summarize, various cancer-related endpoints have been associated with PAH 

6 carcinogenicity. Tumor initiation ability and AhR affinity were shown to correspond well with 

7 complete carcinogenicity, while bacterial mutagenesis was not highly correlated with tumor 

8 formation (Sjogren et al., 1996; Lavoie et al., 1979). DNA adduct formation corresponded with 

9 lung adenoma formation in A/J mice for several PAHs (Sjogren et al., 1996; Ross et al., 1995; 

10 LaVoie et al., 1979). The development of RPFs in this analysis considered both tumorigenicity 

11 and cancer-related endpoints (e.g., mutagenicity, clastogenicity, morphological transformation). 

12 

13 2.7. SIMILARITIES IN RELATIVE POTENCY ESTIMATES ACROSS SPECIES AND 

14 EXPOSURE ROUTES 

15 Available studies suggest that the potency of individual PAHs is generally consistent 

16 across species and study protocols. The consistency of potency estimates based on in vivo 

17 tumorigenicity studies conducted using different study protocols and exposure routes in varying 

18 species/strains of test animals is summarized below. 

19 Nisbet and LaGoy (1992) and Clement Associates (1988) reported that RPFs for PAHs 

20 are reasonably consistent across different study protocols using varying species/strains of 

21 laboratory animals. RPF estimates were calculated in multiple test systems including mouse skin 

22 complete carcinogenesis studies, mouse skin tumor initiation studies, studies in rat lung 

23 (implantation), other rat studies (intrapulmonary injection, subcutaneous injection), and newborn 

24 mouse (intraperitoneal injection). The RPF estimates for specific PAHs calculated from 

25 different assay systems varied by less than an order of magnitude. The relative potency of 

26 individual PAHs to benzo[a]pyrene was also shown to be very similar when based on data in 

27 different strains of mice using different mouse tumor initiation models (Slaga and Fisher, 1983). 

28 Muller et al. (1997) compared the relative potency of benzo[a]pyrene and 3-methylcholanthrene 

29 from data generated in three species (rat, mouse, and hamster). Similar RPF values (i.e., within a 

30 factor of 2) were derived for oral exposures in mice, rats, and hamsters. In their comparison 

31 across different exposure routes (oral, respiratory, and dermal), Muller et al. (1997) reported 

32 similar relative potencies for benzo[a]pyrene and 3-methylcholanthrene (within a factor of 2) for 

33 data from rats exposed via oral and respiratory routes, and for mice exposed via oral and dermal 

34 routes. The relative potency for respiratory exposure in mice was an order of magnitude lower 

35 than relative potencies for the other two exposure routes. 

36 Schneider et al. (2002) performed a more recent evaluation of the impact of exposure 

37 route on the determination of RPFs. Potency ratios were calculated for several carcinogenicity 

38 bioassays by dividing the carcinogenic potency of a PAH mixture by the carcinogenic potency of 
46 DRAFT – DO NOT CITE OR QUOTE

1 benzo[a]pyrene as a single substance. The potency ratios were observed to vary by exposure 

2 route and target organ. For example, potency ratios associated with forestomach tumors from 

3 oral exposure ranged from 0.7 to 1.2 (i.e., the potencies of the PAH mixtures and benzo[a]pyrene 

4 to induce forestomach tumors were approximately equal). This suggested that these tumors may 

5 be attributable to the benzo[a]pyrene content of the mixture. Potency ratios for skin tumor 

6 production from dermal exposure ranged from 2 to 11, whereas RPFs calculated for lung tumors 

7 from oral exposure, pulmonary implantation, or inhalation were greater than 20. These results 

8 suggested that the benzo[a]pyrene content of PAH mixtures may be only slightly responsible for 

9 lung and dermal carcinogenicity. Schneider et al. (2002) suggested that RPF estimates should be 

10 derived separately for oral, dermal, and inhalation exposure using studies with the relevant 

11 exposure pathway. 

12 To summarize, there is some consistency within the in vivo carcinogenicity database for 

13 relative potency estimates derived from different species and strains exposed by various routes, 

14 although this is an area for which further research is needed. However, Schneider et al. (2002) 

15 have cautioned that potency ratios appear to cluster by exposure route and target organ and have 

16 suggested that route-specific RPFs be developed. There is also some concern regarding the use 

17 of benzo[a]pyrene as an index chemical to estimate lung cancer from PAH mixtures, considering 

18 that the lung is relatively insensitive to benzo[a]pyrene-induced tumorigenicity following oral 

19 exposure (Gaylor et al., 1998). Section 8.6 provides a comparison of RPF values calculated in 

20 this report, using bioassay data from different exposure routes and study designs. RPF values 

21 were comparable across most exposure routes, with the exception of the newborn mouse 

22 intraperitoneal injection studies. 

23 

24 2.8. DOSE ADDITIVITY OF PAHs IN COMBINED EXPOSURES

25 Use of the RPF approach assumes that doses of component chemicals that act in a similar 

26 manner can be added together, after scaling the potencies relative to the index chemical, and that 

27 interaction effects do not occur (U.S. EPA, 2000, 1986). The level of confidence in the RPF 

28 approach is increased if additivity can be demonstrated experimentally, even with simple 

29 mixtures. For PAHs, the assumption of additivity cannot be confirmed or refuted based on the 

30 available experimental data. It appears that risks may be generally additive for complex 

31 mixtures, while binary mixtures can exhibit antagonism, synergism, or additivity as discussed 

32 below. 

33 The complexity of potential interactions for tumorigenesis of binary mixtures of PAHs is 

34 illustrated in Table 2-2. The nature of the interaction varies with the PAHs evaluated and the 

35 study conditions (e.g., vehicle used, dose selection, study method). Many studies were designed 

36 to evaluate the combined administration of a known carcinogen with either a weak carcinogen or 

37 a noncarcinogenic PAH. The true nature of the interaction (i.e., additive, synergistic, or 

38 antagonistic) can be difficult to determine in studies wherein the tumorigenic response is not 
47 DRAFT – DO NOT CITE OR QUOTE

1 measured for both PAHs given alone and in combination. These studies can distinguish between 

2 an enhanced or cocarcinogenic response and an inhibitory response, but a further classification 

3 cannot be made. The interactions described as cocarcinogenic in Table 2-2 may be either 

4 additive or synergistic in nature. 

5 

Table 2-2. Studies of binary mixtures of PAHs and tumorigenicity 

Reference Endpoint Findings Net effect 

Cavalieri et al., 1983 Mouse skin 

carcinogenicity 

BaP and CPcdP given together resulted in 

synergistic effect at low and intermediate doses; 

three- to sevenfold increase in relative risk at 

intermediate dose of both BaP and CPcdP as 

compared to the sum of the relative risk for the 

same dose of each PAH given alone. 

S 

DiGiovanni et al., 1982 Skin tumor initiation in 

mice 

BeP increased BaP tumor initiation (30%↑), 

inhibited tumor initiation by DMBA (84%↓) and 

DBahA (48%↓) and produced no change in 

combination with 3-MC; DBacA inhibited tumor 

initiation by DMBA (92%↓), DBahA (39%↓), and 

3-MC (61%↓) and produced no change in 

combination with BaP. 

Co, I 

Falk et al., 1964 Sarcoma induction in 

mice by subcutaneous 

injection 

PH inhibited tumor response of DBahA in ethyl 

laurate vehicle (approximately 30%↓, estimated 

from graph); tumor response was enhanced in 

triethylene glycol vehicle (approximately 50%↑ to 

100% tumor-bearing animals, estimated from 

graph). 

Co, I 

Lavik et al., 1942 Mouse skin tumors 3-MC and BaP, DBahA, or BaA essentially 

additive. 

A 

Pfeiffer, 1973 Sarcoma induction in 

mice by subcutaneous 

injection 

BaP and DBahA less than additive; tumor response 

for combined treatment was within 10% of DBahA 

response. 

I 

Slaga et al., 1979 Skin tumor initiation in 

mice 

BeP, Pyr, or FA increased skin tumor initiation by 

BaP (30, 35, and 23%↑, respectively); BeP, Pyr, or 

FA decreased skin tumor initiation by DMBA (84, 

50, and 34%↓, respectively). 

Co, I 

Steiner, 1955; Steiner 

and Falk, 1951 

Sarcoma induction in 

mice by subcutaneous 

injection 

DBahA and 3-MC in combination roughly additive; 

BaA and CH in combination resulted in synergistic 

effect (9%↑ above additive response); BaA and 

DBahA in combination resulted in inhibition (48%↓ 

below additive response). 

A, S, and I 

Van Duuren and 

Goldschmidt, 1976; 

Goldschmidt et al., 

1973 

Mouse skin 

carcinogenicity 

BeP, BghiP, Pyr, or FA and BaP increased tumors 

over BaP alone (>50% increase in incidence, also 

↑multiplicity); no tumors were observed for PAHs 

without BaP. 

S 

Van Duuren et al., 

1973 

Mouse skin 

carcinogenicity 

BaP and BghiP had cocarcinogenic effect (23%↑ 

over BaP response alone). 

Co 

Warshawsky et al., 

1993 

Mouse skin 

carcinogenicity 

Nontumorigenic dose of BaP increased tumor 

incidence produced by CH (16%↑), anthracene 

(8%↑), and FA (8%↑). 

S 

3-MC = 3-methylchloanthrene; A = additive; Co = cocarcinogenic (enhanced tumorigenicity, study design does not 

allow for determination of A or S); DMBA = 7,12-dimethyl-benz[a]anthracene; I = inhibitory; S = synergistic
48 DRAFT – DO NOT CITE OR QUOTE

1 

2 Slooff et al. (1989) reviewed the available data addressing the carcinogenicity of 

3 individual PAHs and in combination. It was concluded that a generally additive effect was 

4 observed following administration of more than two different PAHs in weight ratios similar to 

5 those occurring in ambient air or in various emissions. Combinations of only two PAHs 

6 produced either additive, synergistic, or inhibitory effects. The complexity of the interaction 

7 among single PAH compounds is thought to be related to effects on metabolic enzyme systems 

8 including induction processes and competitive inhibition. The generlaly additive response noted 

9 for a more complex mixture may reflect the balance between inhibitory and synergistic 

10 processes. 

11 Additivity has been observed in carcinogenicity studies of complex mixtures of PAHs. 

12 Schmähl et al. (1977) evaluated the production of skin tumors following combined dermal 

13 treatment with 11 PAHs found as constituents of automobile exhaust. Tumor findings were 

14 presented separately for two groups of PAHs. High potency carcinogens (Group 1) included 

15 benzo[a]pyrene, dibenz[a,h]anthracene, benz[a]anthracene, and benzo[b]fluoranthene. Lower 

16 potency PAHs (Group 2) included anthracene, benzo[e]pyrene, benzo[g,h,i]perylene, chrysene, 

17 fluoranthene, phenanthrene, and pyrene. Chronic dermal exposure to PAHs in both groups 

18 resulted in an additive response when compared to the tumor response for each group alone. 

19 Nesnow et al. (1998b) evaluated lung tumor formation in A/J mice following combined 

20 administration of five carcinogenic PAH compounds (benzo[a]pyrene, benzo[b]fluoranthene, 

21 dibenz[a,h]anthracene, 5-methylchrysene, and cyclopenta[c,d]pyrene). High and low doses were 

22 selected for each PAH in this study based on toxicity, survival, range of response, and predicted 

23 tumor yield. The ratio of PAH doses was designed to simulate PAH ratios found in 

environmental air and emissions samples. PAHs were administered to mice in a 2524 factorial 

25 study design yielding 32 dose groups (combination of five PAHs at high and low doses). The 

26 formation of lung adenomas was evaluated 8 months following intraperitoneal injection of PAH 

27 mixtures. A response surface model was used to evaluate specific interactions among PAHs. 

28 The results of the study indicated that greater-than-additive effects were seen at low doses, while 

29 less-than-additive effects were observed at high doses. However, the magnitude of the 

30 interactions was relatively small (twofold), suggesting that potential interactions are limited in 

31 extent. 

32 Dermal application of binary mixtures of PAHs has also been shown to produce additive, 

33 synergistic, and inhibitory effects on DNA binding in mouse skin (Hughes and Phillips, 1993, 

34 1990). Hermann (1981) demonstrated that many PAHs could both enhance and inhibit the 

35 bacterial mutagenicity of benzo[a]pyrene depending on the relative concentrations in the binary 

36 mixture. Binary mixtures of benzo[a]pyrene and benzo[e]pyrene produced a synergistic 

37 response in the TA98 strain of S. typhimurium (which detects frameshift mutations), and 

38 antagonistic and additive effects in strain TA100 (which detects a broad spectrum of mutations) 
49 DRAFT – DO NOT CITE OR QUOTE

1 depending on the concentration (Hass et al., 1981). Binary mixtures of PAHs have also been 

2 shown to produce antagonistic or less than additive effects in the Ames assay of bacterial 

3 mutagenicity (Barrai et al., 1992; Salamone et al., 1979a). Vaca et al. (1992) demonstrated an 

4 additive effect for sister chromatid exchange induction by combined administration of 

5 benzo[a]pyrene and fluoranthene in human peripheral lymphocytes cocultured with 

6 PCB-induced rodent liver cells. 

7 The effects of binary PAH mixtures on gene expression, DNA adduct formation, 

8 apoptosis, and cell cycle are additive compared to the effects of the individual compounds in 

9 human hepatoma cells (HepG2) (Staal et al., 2007). Equimolar and equitoxic mixtures of 

10 benzo[a]pyrene with either dibenzo[a,l]pyrene, dibenz[a,h]anthracene, benzo[b]fluoranthene, 

11 fluoranthene, or 1-methylphenanthrene were studied. PAH mixtures showed an additive effect 

12 on apoptosis and on cell cycle blockage. The effects of binary mixtures of PAHs on gene 

13 expression were generally additive or slightly antagonistic. 

14 Additivity has also been observed for the mutagenicity of PAHs administered as a 

15 complex mixture (Bostrom et al., 1998; Kaden et al., 1979). Kaden et al. (1979) evaluated the 

16 bacterial mutagenicity of the PAH fraction of kerosene soot using resistance to 8-azaguanine as a 

17 genetic marker for forward mutation in S. typhimurium. Approximately half of the PAHs tested 

18 (34 of 70) produced a significant increase in the mutant fraction in this assay system. The 

19 mutagenicity of the complex soot mixture was demonstrated to be approximately equal to the 

20 additive mutagenicity of the individual components. Bostrom et al. (1998) reported additivity in 

21 the Ames test of bacterial mutagenesis (i.e., reversion to histidine independence) for a mixture of 

22 four PAHs (benzo[a]pyrene, benz[a]anthracene, fluorene, and pyrene) using four different strains 

23 of S. typhimurium.

24 Mechanistic studies have suggested that the outcome of the interaction between two 

25 PAHs in a binary mixture is dependent on changes in metabolism. PAHs can act as both 

26 inducers and competitive inhibitors of the CYP enzymes that are responsible for generation of 

27 reactive metabolites. Benzo[e]pyrene has been shown to alter the oxidative metabolism of 

28 benzo[a]pyrene, which may be related to the cocarcinogenic effect seen in skin tumor initiation 

29 studies (Baird et al., 1984). Alterations in the types and amounts of benzo[a]pyrene metabolites 

30 suggest that benzo[e]pyrene-induced changes may be isozyme specific (Smolarek and Baird, 

31 1984). An increase in the formation of benzo[a]pyrene DNA adducts has also been 

32 demonstrated for coadministration of benzo[e]pyrene in Sencar mouse skin (Smolarek et al., 

33 1987). Fluoranthene and pyrene have been shown to increase the formation of benzo[a]pyrene34 DNA adducts in mouse skin following a combined treatment (Rice et al., 1988, 1984). 

35 Enhancement of the metabolism of benzo[a]pyrene to diol epoxide metabolites and subsequent 

36 DNA binding may explain the increased carcinogenic effect in this case. Phenanthrene did not 

37 increase the formation of benzo[a]pyrene-DNA adducts and was not shown to be cocarcinogenic 

38 following combined administration with benzo[a]pyrene in this study. Cherng et al. (2001) 
50 DRAFT – DO NOT CITE OR QUOTE

1 demonstrated that benzo[g,h,i]perylene increased the formation of benzo[a]pyrene adducts in 

2 hepatoma cells (HepG2) by enhancing benzo[a]pyrene induction of CYP1A1. Benzo[g,h,i]

3 perylene increased the nuclear accumulation of the AHR and/or the activation of the AhR to a 

4 DNA-binding form (Cherng et al., 2001). Benzo[k]fluoranthene altered the metabolic profile of 

5 benz[a]anthracene by increasing the activity of CYP1A1 (Schmoldt et al., 1981). The bacterial 

6 mutagenicity of benz[a]anthracene was enhanced by use of a rodent liver S9 that was obtained 

7 from animals previously exposed to other PAHs (Norpoth et al., 1984). Coadministration of 

8 benzo[a]pyrene and benz[a]anthracene to hamster embryo cell cultures resulted in a decrease in 

9 the metabolism of benzo[a]pyrene, a decrease in the level of DNA binding, and a decrease in 

10 mutation frequency in hamster V79 cells (Smolarek et al., 1986). 

11 In summary, combined administration of binary mixtures of PAHs can result in several 

12 types of joint action (i.e., additive, synergistic, or antagonistic). The nature of the joint action 

13 appears to be dependent on the characteristics of the individual PAHs, related changes in 

14 metabolism and possibly the test species/strain. PAHs can act as both inducers and competitive 

15 inhibitors of the CYP enzymes that are responsible for generation of reactive metabolites. 

16 Additivity has been observed for some complex mixtures of PAHs, suggesting a balance in the 

17 relative metabolism of individual PAHs. For the purposes of this analysis, an assumption is 

18 made that the combination of individual PAHs results in additive effects. Additional research is 

19 needed to characterize the validity of this assumption. 

20 

21 

22 3. DISCUSSION OF PREVIOUSLY PUBLISHED RPF APPROACHES 

23 

24 

25 There are multiple analyses available for the derivation of relative potency estimates for 

26 individual PAHs. All of these analyses utilize benzo[a]pyrene as the index chemical. Table 3-1 

27 compares relative cancer potency values for PAHs presented by several authors. A review of the 

28 derivation of these relative potency values follows. 

29 

30 
51 DRAFT – DO NOT CITE OR QUO

1 

Table 3-1. Comparison among various relative potency estimates for PAHs from the published literature and 

regulatory agencies (1984–2004) 

PAH Abbr 

U.S. 

EPA 

(1993)

Chu 

and 

Chen 

(1984) 

Clement 

(1988) 

Clement 

(1990) 

Rugen et 

al. (1989)

Slooff et 

al. (1989)

Kroese 

et al. 

(2001) 

Nisbet 

and 

LaGoy 

(1992) 

Malcolm 

and 

Dobson 

(1994) 

Meek 

et al. 

(1994)

Muller 

et al. 

(1997) 

Larsen and 

Larsen 

(1998) 

Collins 

et al. 

(1998) 

Ca

for

EP

(20

Acenaphthene AN 0.001 0.001 Acenaphthylene ANL 0.001 0.001 Anthanthrene AA 0.32 0.316 0.28 0.3 Anthracene AC 0 0 0.01 0.01 0.0005 Benzo[a]pyrene BaP 1 1 1 1 1 1 1 1 1 1 1 1 1 Benz[a]anthracene BaA 0.1 0.013 0.145 0.004–

0.006 

0–0.04 <0.1 0.1 0.1 0.014 0.005 0.1 

Benzo[b]fluoranthene BbF 0.1 0.08 0.14 0.1228 0.0235 0.1 0.1 0.06 0.11 0.1 0.1 0.6Benzo[c]phenanthrene BcPH 0.023 0.023 Benzo[e]pyrene BeP 0.004 0.007 0.01 0 0.002 Benzo[g,h,i]perylene BghiP 0.022 0.0212 0.01–0.03 0.03 0.01 0.01 0.012 0.02 Benzo[j]fluoranthene BjF 0.061 0.0523 0.0763 0.1 0.05 0.045 0.05 0.1 0.Benzo[k]fluoranthene BkF 0.01 0.004 0.066 0.0523 0.03–0.09 <0.1 0.1 0.1 0.04 0.037 0.05 0.1 Chrysene CH 0.001 0.001 0.0044 0.05–0.89 0.1–0.03 0.01 0.01 0.026 0.03 0.01 0.Coronene CO 0.001 Cyclopenta[c,d] 

pyrene 

CPcdP 0.023 0.1 0.012 0.02 

Dibenzo[a,h] anthracene 

DBahA 1 0.69 1.11 0.599 5 1 0.89 1.1 

Dibenz[a,c]anthracene DBacA 0.1 Dibenzo[a,e]pyrene DBaeP 0.2 1 Dibenzo[a,h]pyrene DBahP 1.2 1 10 1Dibenzo[a,i]pyrene DBaiP 1.1 0.1 10 1Dibenzo[a,l]pyrene DBalP 1 10 Fluoranthene FA 0–0.06 0.01 0.001 0.001 0.05Fluorene FE 0.001 0.001 Indeno[1,2,3-c,d]pyrene 

IP 0.1 0.017 0.232 0.278 0.00599 0–0.08 0.1 0.1 0.1 0.12 0.067 0.1 0.1 

Perylene Pery 0.001 
52 DRAFT – DO NOT CITE OR QUO

Table 3-1. Comparison among various relative potency estimates for PAHs from the published literature and 

regulatory agencies (1984–2004) 

PAH Abbr 

U.S. 

EPA 

(1993)

Chu 

and 

Chen 

(1984) 

Clement 

(1988) 

Clement 

(1990) 

Rugen et 

al. (1989)

Slooff et 

al. (1989)

Kroese 

et al. 

(2001) 

Nisbet 

and 

LaGoy 

(1992) 

Malcolm 

and 

Dobson 

(1994) 

Meek 

et al. 

(1994)

Muller 

et al. 

(1997) 

Larsen and 

Larsen 

(1998) 

Collins 

et al. 

(1998) 

Ca

for

EP

(20

Phenanthrene PH 0.01 <0.01 0.001 0.001 0.00064 0.0005 Pyrene Pyr 0.081 0.001 0.001 0 0.001 Abbr = abbreviation 
53 DRAFT – DO NOT CITE OR QUOTE

1 U.S. EPA (1993) presented RPFs (termed EOPPs) for seven PAHs (benzo[a]pyrene, 

2 benz[a]anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene, chrysene, 

3 dibenz[a,h]anthracene, indeno[1,2,3-c,d]pyrene) as Provisional Guidance for the risk evaluation 

4 of PAHs at hazardous waste sites. On IRIS (U.S. EPA, 2009), all seven of these compounds 

5 were assigned a cancer weight of evidence classification of Group B2 (probable human 

6 carcinogen, based on sufficient evidence of carcinogenicity in animals) under the U.S. EPA 

7 (1986) Guidelines for Carcinogen Risk Assessment. U.S. EPA (1993) indicated that the data for 

8 PAHs did not meet the criteria for the development of toxicity equivalency factors (TEFs). In 

9 particular, the existing database was limited primarily to studies of metabolism, genotoxicity, 

10 and cancer, and the assumption of additivity was not proven or refuted. The EOPP terminology 

11 was used because this approach was limited to skin painting data and was based on 

12 benzo[a]pyrene exposure from a single (oral) pathway (for the derivation of the slope factor). 

13 This analysis considered only a small subset of PAHs routinely measured in PAH mixtures at 

14 hazardous waste sites. The EOPP values were based on previous evaluations conducted by Chu 

15 and Chen (1984) and Clement Associates (1988) and were calculated for various test systems 

16 (i.e., mouse skin carcinogenesis, subcutaneous injection in mice, intrapulmonary administration 

17 to rats, tumor initiation on mouse skin, and intraperitoneal injection in newborn mice) (Clement 

18 Associates, 1988). Various statistical methods for combining data sets were considered; 

19 however, final EOPP values were based on a single test system (skin painting) and were rounded 

20 to the closest order of magnitude. The EOPPs were recommended for the oral exposure route 

21 only, because the quantitative dose-response assessment for benzo[a]pyrene was from an oral 

22 carcinogenicity bioassay (i.e., an oral cancer slope factor). This recommendation was, however, 

23 complicated by the fact that the EOPPs were derived from comparisons based on dermal 

24 exposure. 

25 Chu and Chen (1984) presented RPF values for the seven PAH compounds described in 

26 the Provisional Guidance described above (U.S. EPA, 1993) (see Table 3-1). These values were 

27 calculated using mouse skin painting data only. Tumor incidence data were modeled using the 

28 linearized multistage model and the resulting ED10 and q1* (upper confidence limit of the linear 

29 slope) were presented for target PAHs and benzo[a]pyrene. The RPFs listed in Table 3-1 

30 represent the ratio of the q1* value for a PAH compound to the q1* value for benzo[a]pyrene 

31 (i.e., q1*PAH ÷ q1*BaP). 

32 Clement Associates (1988) identified 11 published studies that concurrently compared 

33 the carcinogenicity of benzo[a]pyrene with one or more other PAHs, and used the data to derive 

34 relative cancer potencies for 13 PAHs, including benzo[a]pyrene. Test protocols used in this 

35 analysis included mouse skin complete carcinogenesis, initiation-promotion on mouse skin, 

36 subcutaneous injection into mice, lung implantation in rats, and intraperitoneal injection into 

37 newborn mice. Tumor incidence data were fit to a simplified version of the Moolgavkar38 Venson-Knudsen (MVK) two-stage model and to the linearized multistage model to obtain low-
54 DRAFT – DO NOT CITE OR QUOTE

1 dose cancer potency values (transition rates and low-dose slope factors, respectively). Most of 

2 the estimates were derived using data for multiple exposure levels and controls, but some were 

3 based on a single exposure level and a control. RPFs were calculated as the ratio of the 

4 estimated transition rate or slope factor for a particular PAH to the corresponding values for 

5 benzo[a]pyrene from the same study. Clement Associates (1988) selected representative RPFs 

6 for each of the studied PAHs based on evaluations of the quality of the studies from which the 

7 estimates were obtained. 

8 Clement Associates (1990) also derived relative cancer potencies for eight PAHs based 

9 on tumor incidence data from rat lung implantation data only (Deutsch-Wenzel, 1983). The data 

10 were restricted to a single group of studies using a defined experimental protocol in order to 

11 address issues of questionable data quality associated with other studies. Data quality concerns 

12 cited for other studies include variation in survival, saturation of the carcinogenic effect, 

13 outmoded pathological classification, and inadequate controls. The RPF values based on rat lung 

14 implantation data were comparable to those originally derived by Clement Associates (1988) 

15 (see Table 3-1). 

16 Rugen et al. (1989) proposed a relative potency approach to establish acceptable 

17 exposure levels (AELs) for six carcinogenic PAHs in drinking water (listed in Table 3-1). These 

18 authors reviewed mouse skin painting studies in which the cancer potency of benzo[a]pyrene 

19 was compared with those of other PAHs (Bingham and Falk, 1969; Wynder and Hoffmann, 

20 1961, 1959a, b). The following relationship was used to calculate conversion factors (“relative 

21 tumor dose” = RTD) to derive AELs for these PAHs from the AEL for benzo[a]pyrene: RTD = 

22 (d1/n1)/(d2/n2); where d1 and n1 represented a dosage level and associated tumor incidence after a 

23 given exposure duration to a certain PAH, PAH1, and d2 and n2 represented similar quantities for 

24 exposure to the index PAH, benzo[a]pyrene, for the same exposure duration. The AEL for a 

particular PAH was then derived with the following relationship: AEL(PAHi) 25 = AEL(benzo[a]pyrene) ×

RTD(PAHi) 26 . In this approach, RTDs for PAHs more potent than benzo[a]pyrene were less 

27 than 1 and RTDs for PAHs less potent than benzo[a]pyrene were greater than 1. The reciprocal 

28 of the RTDs derived by Rugen et al. (1989) were comparable to the RPFs presented by other 

29 authors and are presented as such in Table 3-1. 

30 The Netherlands (RIVM) proposed RPF values for 10 PAHs (naphthalene, anthracene, 

31 phenanthrene, fluoranthene, chrysene, benz[a]anthracene, benzo[k]fluoranthene, benzo[a]pyrene, 

32 benzo[g,h,i]perylene, and indeno[1,2,3-c,d]pyrene) (Slooff et al., 1989). RPFs were calculated 

33 as a ratio of ED50 values that were calculated using a simple linear model. For dermal studies in 

34 which the latency period was determined, the tumor incidence was divided by latency and 

35 concentration, and the values were averaged for the different concentrations. Kroese et al. 

36 (2001) provided an update of the RPF values calculated by Slooff et al. (1989) by incorporating 

37 more recent evaluations conducted by other authors (Larsen and Larsen, 1998; Nesnow et al., 
55 DRAFT – DO NOT CITE OR QUOTE

1 1998b; Muller, 1997; Nisbet and LaGoy, 1992). The RPF values for chrysene and fluoranthene 

2 were decreased, while other values remained similar to those originally proposed (see Table 3-1). 

3 Nisbet and LaGoy (1992) proposed toxicity equivalence factors for 17 PAHs commonly 

4 found at hazardous waste sites. These authors reviewed published studies in which the 

5 tumorigenic potencies of one or more PAHs were compared with benzo[a]pyrene (essentially the 

6 same as those reviewed by Clement Associates, 1988) and rounded, to an order of magnitude, the 

7 estimates presented by Clement Associates (1988) for seven carcinogenic PAHs 

8 (dibenz[a,h]anthracene, benz[a]anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene, 

9 indeno[1,2,3-c,d]pyrene, benzo[g,h,i]perylene, and chrysene) (see Table 3-1). Nisbet and LaGoy 

10 (1992) argued that the rounded estimates more accurately reflected the uncertainty in the 

11 estimates than the values presented by Clement Associates (1988). Nisbet and LaGoy (1992) 

12 stated that Clement Associates (1988) proposed a TEF of 0.32 for anthracene (CASRN 

13 120-12-7), but examination of the original report shows that Clement Associates (1988) 

14 proposed this value for anthanthrene (CASRN 191-26-4) and did not propose a value for 

15 anthracene. Nisbet and LaGoy (1992) assigned a value of 0.01 to anthracene. In addition, 

16 Nisbet and LaGoy (1992) arbitrarily assigned TEFs of 0.001 to eight other PAHs for which 

17 adequate evidence of carcinogenicity in animals was not available (acenaphthene, 

18 acenaphthylene, fluoranthene, fluorene, 2-methylnaphthalene, naphthalene, phenanthrene, and 

19 pyrene). In defense of this assignment, the argument was made that some of these PAHs have 

20 been shown to have some, albeit limited, evidence for carcinogenic or genotoxic activity in some 

studies (e.g., phenanthrene and naphthalene221 ). The RPF value proposed for 

22 dibenz[a,h]anthracene was substantially higher than that proposed by Clement Associates (1988). 

23 Nisbet and LaGoy (1992) indicate that their analysis of the dose-response data suggests that an 

24 RPF value of 5 is more appropriate for environmental exposures where the chemically-related 

25 tumor incidence rate would be approximately <25%. 

26 Malcolm and Dobson (1994) used RPFs for 23 PAHs to calculate environmental 

27 assessment levels (EALs) for atmospheric PAHs (sponsored by the Great Britain Department of 

28 the Environment). The RPFs were derived from previously reported review papers (Nisbet and 

29 LaGoy, 1992; Rugen et al., 1989; Clement Associates, 1988; Chu and Chen, 1984), as well as the 

30 primary literature describing pulmonary implant, skin painting, subcutaneous injection, and 

31 mouse skin DNA binding studies. No information was provided regarding the methodology used 

32 to derive RPFs from specific experimental studies. The proposed RPF values for individual 

33 PAHs were the highest values reported in the literature. Many of the RPF values are similar to 

34 those reported by Nisbet and LaGoy (1992). RPFs were additionally reported for 

35 benzo[e]pyrene, coronene, cyclopenta[c,d]pyrene, dibenz[a,c]anthracene, and perylene. The 

36 benzo[e]pyrene and cyclopenta[c,d]pyrene RPFs were apparently calculated directly from mouse 

 

2

It should be noted that a recent bioassay for naphthalene has shown increased incidence of nasal tumors in exposed 

rats (NTP, 2000). 
56 DRAFT – DO NOT CITE OR QUOTE

1 skin painting studies (Habs et al., 1980; Hoffmann and Wynder, 1966; Wynder and Hoffmann, 

2 1959a, b). Coronene and perylene were arbitrarily assigned RPF values of 0.001 given the 

3 International Agency for Research on Cancer (IARC) and U.S. EPA designation as “not 

4 classifiable as to human carcinogenicity” (similar approach to Nisbet and LaGoy, 1992). 

5 Dibenz[a,c]anthracene was assigned an RPF value of 0.1 based on the IARC designation of 

6 “possibly carcinogenic to humans.” 

7 Health Canada (Meek et al., 1994) proposed RPFs for five PAHs (benzo[a]pyrene, 

8 benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[j]fluoranthene, and indeno[1,2,3-cd]pyrene) 

9 based on the results of multistage modeling of incidence data in Osborne-Mendel rats treated by 

10 lung implantation (Deutsch-Wenzel et al., 1983). Values were based on a comparison of the 

11 doses that caused a 5% increase in tumor incidence (ED05). RPFs were calculated as the ratio of 

12 the ED05 for benzo[a]pyrene to the ED05 for a specific PAH compound. 

13 The Ontario Ministry of Environment and Energy (Muller et al., 1997) proposed RPF 

14 values for 209 PAHs using data from dermal studies in mouse skin or rat lung bioassays. Most 

15 of these PAHs were alkylated PAHs, PAH metabolites, or heterocyclic PAH compounds. The 

16 17 unsubstituted PAHs that were evaluated in this analysis are listed in Table 3-1. Muller et al. 

17 (1997) derived a standard time of observation in order to account for varying study duration 

18 across experiments. Several dose-response models were considered for the evaluation of tumor 

19 incidence and multiplicity, and linear regression was selected as the preferable method. 

20 Tumorigenic potency (i.e., the slope of incidence/mg) was determined separately for each data 

21 set based on the following order of preference regarding study type: tumor initiation in 

22 CD-1 mice, tumor initiation in SENCAR mice, rat lung implantation, and complete 

23 carcinogenicity in C57BL mice. RPFs were determined as the ratio of PAH potency to the 

24 potency of benzo[a]pyrene. RPF values derived by Muller et al. (1997) were comparable to 

25 values estimated by other authors. 

26 Larsen and Larsen (1998) estimated RPFs for 23 PAHs based on a compilation of 

27 available carcinogenicity data in animals using oral, pulmonary, and skin application of PAHs. 

28 The authors indicated that these values represent an entirely subjective estimate of relative 

29 potency; however, further detail regarding the derivation of RPF estimates was not provided. 

30 Collins et al. (1998) developed RPFs (termed potency equivalency factors [PEFs]) for 

31 21 PAHs, 10 of these were either methyl- or nitro-substituted or heterocyclic PAHs. A hierarchy 

32 of data types was utilized to provide an order of preference for data utilization in calculating 

33 RPFs. Because the analysis focused on PAHs as air contaminants, tumor data from inhalation 

34 studies were preferred (although none were found), followed by intratracheal or intrapulmonary 

35 instillation, oral administration, skin-painting, and subcutaneous or intraperitoneal injection. 

36 Genotoxicity and structure activity data were considered the least-preferred data type for 

37 calculation of RPFs. Collins et al. (1998) noted that a wide range of PEFs were observed for 

38 individual chemicals using different types of data (e.g., mutagenicity versus tumor data). The 
57 DRAFT – DO NOT CITE OR QUOTE

1 basis for the derivation of individual RPF values was presented in a California EPA (2002) 

2 technical support document. RPF values for benz[a]anthracene, benzo[b]fluoranthene, 

3 benzo[j]fluoranthene, benzo[k]fluoranthene, indeno[1,2,3-c,d]pyrene, and chrysene were similar 

4 to those described by Clement Associates (1988). Additional RPFs for dibenzo[a,e]pyrene, 

5 dibenzo[a,h]pyrene, dibenzo[a,i]pyrene, and dibenzo[a,l]pyrene were calculated using mouse 

6 skin and rat mammary gland data (Cavalieri et al., 1991, 1989). A cancer slope factor was 

7 directly calculated for dibenz[a,h]anthracene using the tumor incidence data from a drinking 

8 water study (Snell and Stewart, 1962). The relative potency of dibenz[a,h]anthracene was 

9 estimated to be 0.1, when compared to the oral potency for benzo[a]pyrene. 

10 Revised California EPA RPFs were recently developed for benzo[b]fluoranthene, 

11 benzo[j]fluoranthene, chrysene, dibenzo[a,h]pyrene, and dibenzo[a,i]pyrene (California EPA, 

12 2004). Cancer potency estimates were derived from lung adenoma data in newborn mice treated 

by intraperitoneal injection. Potency estimates represented the upper 95th 13 percent confidence 

14 limit on the linear term of the multistage model fit for the newborn mouse dose-response data. 

15 Because benzo[a]pyrene was demonstrated to be 75 times more toxic in newborn mouse 

16 intraperitoneal assays than in adult oral studies, oral equivalent potencies for individual PAHs 

17 were derived by adjusting the cancer potency downward by a factor of 75. The RPFs listed in 

18 Table 3-1 were calculated as the ratio of the oral equivalent potency for a PAH to the oral 

19 potency estimate for benzo[a]pyrene. This methodology resulted in a significant increase in RPF 

20 values for benzo[b]fluoranthene, benzo[j]fluoranthene, and chrysene when compared with other 

21 approaches. 

22 In summary, several approaches are available for the determination of RPFs for PAHs. 

23 RPF values are proposed in at least one study for a total of 27 PAHs (see Table 3-1). Because 

24 these approaches generally rely on similar bioassay data and modeling methods, the resulting 

25 RPF values are fairly comparable for most PAHs across studies. Reports by Larsen and Larsen 

26 (1998) and Malcolm and Dobbs (1994) did not provide sufficient information on the 

27 methodology used to calculate RPF estimates and are therefore more uncertain. Variable RPF 

28 estimates were reported for benz[a]anthracene, chrysene, and indeno[1,2,3-c,d]pyrene. RPF 

29 values were also highly variable for dibenzo[a,e]pyrene, dibenzo[a,h]pyrene, dibenzo[a,i]pyrene, 

30 and dibenzo[a,l]pyrene; however, these were only presented in a few recent studies. As 

31 indicated above, the recent California EPA (2004) approach to estimating RPFs provides 

32 considerably higher RPF values for benzo[b]fluoranthene, benzo[j]fluoranthene, and chrysene, 

33 compared with other approaches. 

34 U.S. EPA is reevaluating the RPF approach for PAHs in this analysis due to the evolution 

35 of the state of the science and increased understanding of PAH toxicology. A great deal of 

36 scientific research on PAHs has been conducted since the 1993 Provisional Guidance was 

37 developed. Toxicological data are available for a larger number of PAHs and cancer-related 

38 endpoints. However, the database for PAHs still does not meet the criteria for the derivation of 
58 DRAFT – DO NOT CITE OR QUOTE

1 TEFs. U.S. EPA (2000) defines TEFs as special types of RPFs that are derived when there are 

2 abundant data supporting a specific mode of action that is pertinent to all health endpoints. RPFs 

3 may be derived when the mode of action is less certain or is known for only a subset of all health 

4 endpoints. The major differences in the use of TEFs and RPFs is that TEFs are applied to all 

5 health endpoints, exposure routes, and exposure durations (U.S. EPA, 2000), while RPFs may be 

6 limited to specific endpoints, routes, or durations. In the case of PAHs, there are inadequate data 

7 to identify a specific mode of action that is applicable across all health endpoints. Most of the 

8 available toxicological data are limited to cancer endpoints and there are few data on the 

9 potential mode(s) of action for other effects. As a result, the more generalized RPF approach is 

10 considered appropriate for PAHs. 

11 

12 3.1. PREVIOUS EFFORTS TO VALIDATE RPF APPROACH 

13 Several studies have attempted to validate the RPF approach by comparing the cancer 

14 risk of a PAH mixture measured experimentally with the cancer risk that was predicted using the 

15 RPF method (Muller et al., 1997; McClure, 1996; Goldstein et al., 1994; Clement Associates, 

16 1990, 1988; Krewski et al., 1989). These studies provide semi-quantitative information on the 

17 overall uncertainty in using a component-based approach. Consistent findings were not reported 

18 across these studies. Some studies suggested that the RPF approach would closely predict the 

19 cancer risks associated with PAH mixtures while others indicated that cancer risks might be 

20 over- or underestimated. 

21 Clement Associates (1988) evaluated the usefulness of selected RPFs to predict the tumor 

22 incidence observed in a mouse skin painting assay. Schmähl et al. (1977) exposed groups of 

23 mice to multiple doses of benzo[a]pyrene alone or to one of two defined mixtures of PAHs. The 

24 first of these mixtures was comprised of benzo[a]pyrene, dibenz[a,h]anthracene, 

25 benz[a]anthracene, and benzo[b]fluoranthene. The second mixture contained seven PAHs: 

26 phenanthrene, anthracene, fluoranthene, pyrene, chrysene, benzo[e]pyrene, and 

27 benzo[g,h,i]perylene. The predicted tumor incidences for the animals treated with the mixtures 

28 were calculated from benzo[a]pyrene equivalents of the mixture and dose response modeling of 

29 the Schmähl et al. (1977) data for benzo[a]pyrene alone. Predicted tumor incidences for the first 

30 mixture were comparable to observed tumor incidences, while predicted values were greater than 

31 the observed values for the second mixture. 

32 Clement Associates (1990) examined the utility of a relative potency approach, in which 

33 relative cancer potency estimates of eight PAHs were used, to predict the cancer potencies of 

34 each of four complex mixtures containing many PAHs and other substances: gasoline engine 

35 exhaust condensate, flue-gas condensate from coal-fired residential furnaces, diesel engine 

36 exhaust condensate, and sidestream smoke condensate of cigarettes. Relative cancer potencies 

37 (compared to benzo[a]pyrene) for each of the four complex mixtures were calculated using a 

38 simplified version of the MVK two-stage model and tumor incidence data from a series of 
59 DRAFT – DO NOT CITE OR QUOTE

1 published rat lung implantation studies that examined the carcinogenicity of each complex 

2 mixture, various sub-fractions of the mixtures, and benzo[a]pyrene (Grimmer et al., 1988, 

3 1987a, b, 1984). Lung implantation data (Deutsch-Wenzel, 1983) were used to calculate RPFs 

4 for benzo[b]fluoranthene, benzo[e]pyrene, benzo[j]fluoranthene, benzo[k]fluoranthene, 

5 indeno[1,2,3-c,d]pyrene, anthanthrene, benzo[g,h,i]perylene, and benzo[a]pyrene. The sum of 

6 the benzo[a]pyrene exposure equivalents for the eight PAHs (i.e., the sum of the products of the 

7 relative cancer potencies of the eight PAHs multiplied by their concentrations in the respective 

8 complex mixtures) accounted for only minor fractions of the total carcinogenicity of each of the 

9 four complex mixtures. When the assumption was made that each of the eight PAHs was as 

10 potent as benzo[a]pyrene, the sum of the benzo[a]pyrene equivalents still accounted for only 

11 minor fractions of the carcinogenicity of each mixture. Clement Associates (1990) concluded 

12 that the cancer risk associated with a complex PAH mixture could not be estimated reliably from 

13 measurements of a few indicator components, and further speculated that the underestimation 

14 occurred because complex mixtures that occur in the environment contain many PAHs that have 

15 not been studied in cancer tests, but which may be carcinogenic. In addition, complex PAH 

16 mixtures found in the environment contain other potential carcinogens including substituted and 

17 heterocyclic PAHs and non-PAH components. 

18 Krewski et al. (1989) compared the observed tumor response rate for two PAH mixtures 

19 in mice with the tumor response predicted using the RPFs for 13 individual PAHs; chemical 

20 characterization of the mixture was not provided. With the exception of the highest dose, the 

21 predicted tumor response for mixture 1 was similar to the observed response. For mixture 2, the 

22 predicted tumor response value was higher than the observed response. 

23 Goldstein et al. (1994) compared the experimental carcinogenicity of a MGP residue to 

24 the predicted cancer risk using the Nisbet and LaGoy (1992) RPF scheme. The RPF method 

25 underestimated the potential carcinogenicity of the mixture. The lack of correspondence was 

26 suggested to be related to the presence of unidentified carcinogens in the mixture or possible 

27 synergistic interactions between PAHs. 

28 McClure et al. (1996) compared the tumor response predicted using U.S. EPA’s 1993 

29 provisional values (i.e., EOPPs) to the observed response reported in studies of mice exposed to 

30 synthetic and complex mixtures of PAHs. The results of this analysis were mixed. EOPP values 

31 closely predicted the mouse tumor response to subcutaneous or dermal application of synthetic 

32 mixtures containing relatively potent carcinogens, while overestimating the response to synthetic 

33 mixtures containing only relatively weak carcinogens (similar to findings of Clement Associates, 

34 1988). Mouse skin tumor initiation with several coal liquids was closely predicted by the EOPP 

35 approach; however, this method underestimated the tumor response from lung implantation of 

36 coal furnace emission condensate and its PAH-containing neutral fraction. 

37 The validation analyses that were performed by Muller et al. (1997) consisted of 

38 component versus whole mixture risk comparisons using data for smoky coal and coke oven 
60 DRAFT – DO NOT CITE OR QUOTE

1 emissions. The human lung cancer risks that were estimated using the RPF approach were 

2 compared to the whole mixture cancer risk derived from epidemiology studies. The relative 

3 content of PAHs (compared to benzo[a]pyrene) in the mixture was determined analytically (for 

4 smoky coal and coke oven emissions) or was estimated as a standard mixture assumed to 

5 represent an average PAH profile. The RPF method produced PAH cancer risk estimates that 

6 were significantly lower than the risk estimates derived from epidemiology studies. 

7 

8 

9 4. EVALUATION OF THE CARCINOGENICITY OF INDIVIDUAL PAHs 

10 

11 

12 4.1. DATABASE OF STUDIES ON PAH CARCINOGENICITY AND CANCER13 RELATED ENDPOINTS 

14 A database of primary literature relevant to the RPF approach for PAHs was developed. 

15 This was accomplished through the following means: 

16 

17 • Definition of the study types that were considered relevant to relative potency 

18 development; 

19 

20 • Review of reference lists from review articles and other secondary sources; 

21 

22 • Identification of selected PAHs to be included in search of open literature; 

23 

24 • Performance of targeted searches of open literature on selected PAHs; and 

25 

26 • Population of the database with references and meaningful keywords. 

27 

28 The study types that were considered most useful for RPF derivation were rodent 

29 carcinogenicity bioassays (all routes) in which one or more PAHs was tested at the same time as 

30 benzo[a]pyrene. In addition, in vivo and in vitro data for cancer-related endpoints (in which one 

31 or more PAHs and benzo[a]pyrene were tested simultaneously) were obtained, including studies 

32 on the formation of DNA adducts, mutagenicity, chromosomal aberrations, aneuploidy, DNA 

33 damage/repair/recombination, unscheduled DNA synthesis, and cell transformation. Although it 

34 would be possible to calculate RPFs from studies where a PAH and benzo[a]pyrene were tested 

35 by the same laboratory using the same test system but at different times, this approach was not 

36 considered because it could introduce differences in the dose-response information that are 

37 unrelated to the chemical (e.g., variability associated with laboratory environment conditions, 

38 animal handling, food supply). Thus, studies in which benzo[a]pyrene was not tested 

39 simultaneously with another PAH were not considered in this analysis. 

40 Studies of AhR binding/activation were not considered for use in deriving RPFs because 

41 there is evidence indicating that highly mutagenic fjord-region PAHs are potent carcinogens, 
61 DRAFT – DO NOT CITE OR QUOTE

1 despite a lower AhR affinity (reviewed by Bostrom et al., 2002). Likewise, some PAHs that 

2 strongly activate the AhR, such as benzo[k]fluoranthene (Machala et al., 2001), are only weakly 

3 carcinogenic. In addition, some studies have demonstrated the formation of DNA adducts in the 

4 liver of AhR knockout mice following intraperitoneal or oral exposure to benzo[a]pyrene 

5 (Sagredo et al., 2006; Uno et al., 2006; Kondraganti et al., 2003). These findings suggest that 

6 there may be alternative (i.e., non-AhR mediated) mechanisms of benzo[a]pyrene activation in 

7 the mouse liver, and the AhR affinity would not be a good predictor of carcinogenic potency. 

8 Several study types were initially excluded from the database because they did not 

9 provide carcinogenicity or cancer-related endpoint information for individual PAHs. These 

10 include biomarker studies measuring DNA adducts in humans, studies of PAH metabolism, and 

11 studies of PAH mixtures. Although these studies contain important information on human 

12 exposure to PAH mixtures and the mode of action for PAH toxicity, they generally do not 

13 contain dose-response information that would be useful for calculation of RPF estimates. In 

14 addition to the primary bioassay and cancer-related endpoint studies described above, the RPF 

15 database also includes information on PAH mode of carcinogenic action, interactions among 

16 PAHs in mixtures, and the influence of exposure route on carcinogenic action of PAHs. 

17 Primary studies were identified through review of available secondary sources and 

18 review articles, supplemented by a targeted literature search. A complete list of the secondary 

19 sources that were reviewed is contained in Appendix A. A literature search strategy was 

20 developed by first constructing a list of the individual PAHs to be included. The list of PAHs 

21 was restricted to unsubstituted PAHs with three or more fused aromatic rings containing only 

22 carbon and hydrogen atoms, because these are the most widely studied members of the PAH 

23 chemical class. Heterocyclic PACs or PAHs with substituted groups (e.g., alkyl, hydroxyl, 

24 sulfhydryl, amino, or nitro groups) were not included. An initial search yielded a list of PAHs 

25 for which toxicological data are available. Individual PAHs were then chosen for the literature 

26 search because they were known to have toxicological information relevant to cancer, and in 

27 most cases, their presence in environmental sources of PAH exposure was known. Using these 

28 criteria and excluding benzo[a]pyrene, 74 PAHs were identified from primary and secondary 

29 sources (see Table 2-1 in Section 2). 

30 A search of the open literature was conducted in the Medline (PUBMED) database for 

31 the PAHs identified. This database encompasses many of the studies that would also be found in 

32 TOXLINE and Cancer Lit (the latter is no longer available as a separate database). Medline 

33 (PUBMED) was searched by CASRN in conjunction with cancer and cancer-related endpoint 

34 keywords. The search was not limited by publication date to ensure that all relevant studies were 

35 identified. A few compounds did not show any result when searched by CASRN. For these 

36 PAHs, an additional search by name was conducted. Search results, including Medline 

37 keywords, were downloaded directly into the working RPF database. 
62 DRAFT – DO NOT CITE OR QUOTE

1 In addition to Medline, computer searches of the following databases and websites were 

2 conducted: IARC, World Health Organization (WHO), Agency for Toxic Substances and 

3 Disease Registry (ATSDR), Health Canada, NTP, California EPA’s Office of Environmental 

4 Health Hazard Assessment (OEHHA), the Substance Registry System, CCRIS, TSCATS, and 

5 DSSTOX. 

6 Primary and secondary studies were entered in the RPF database and relevant keywords 

7 (identifying study type, whether benzo[a]pyrene was included, route of administration, target 

8 organ, etc.) were identified for each study. The list of keywords was developed in order to 

9 facilitate database searching for references on a specific topic. Quality assurance procedures 

10 were employed to ensure that database references were properly keyword-coded for retrieval. 

11 

12 4.2. STUDIES IN HUMANS 

13 Numerous studies have evaluated cancer outcomes in PAH-exposed individuals 

14 (reviewed in Bostrom et al., 2002; WHO, 1998; ATSDR, 1995; IARC, 1987, 1983, 1973). 

15 However, since these exposures were to complex mixtures containing multiple PAH 

16 carcinogens, they did not provide adequate data to evaluate the human carcinogenicity of 

17 individual PAH compounds. Epidemiology studies have focused on occupational exposure to 

18 PAH mixtures. Emissions from coke production, coal gasification, aluminum production, iron 

19 and steel founding, coal tars, coal tar pitches, and soot have produced lung cancer in humans 

20 (Bostrom et al., 2002). Skin and scrotal cancers have resulted from exposure to coal tar, coal tar 

21 pitches, nonrefined mineral oils, shale oils, and soot (Larsen and Larsen, 1998; WHO, 1998; 

22 ATSDR, 1995). Occupational studies clearly demonstrate exposure-response relationships for 

23 PAH mixtures; however, quantitative estimates of risk are limited primarily to lung cancer in 

24 coke oven workers (Bostrom et al., 2002; Larsen and Larsen, 1998; ATSDR, 1995). 

25 Biomonitoring of exposure to PAHs includes measurement of DNA and protein adducts 

26 and measurement of urinary metabolites of PAHs, studies on genetic polymorphisms of CYP450 

27 and other enzymes, and changes in PAH metabolism (Bostrom et al., 2002; Larsen and Larsen, 

28 1998; ATSDR, 1995). While these studies demonstrate the degree of exposure to PAHs from 

29 various settings, quantitative dose-response data for humans exposed to individual PAHs are not 

30 available. Cancer-related endpoint studies that were performed using human cell lines are 

31 presented with similar assays in other mammalian species in Section 4.3. 

32 

33 4.3. STUDIES IN ANIMALS 

34 The database of studies investigating cancer or cancer-related endpoints in animals 

35 exposed to PAHs is extensive. For the purpose of developing relative potency estimates, only 

36 those studies that included at least one selected PAH and benzo[a]pyrene as a reference 

37 compound were reviewed. Studies were excluded if PAH potency comparisons were not 

38 conducted in the same laboratory in concurrent experiments. Studies without benzo[a]pyrene are 
63 DRAFT – DO NOT CITE OR QUOTE

1 listed in two separate bibliographies in Appendix B. Table B-1 of the appendix shows PAHs that 

2 were assayed with or without benzo[a]pyrene. Table B-1 shows that 32 of the 74 PAHs were 

3 assayed with benzo[a]pyrene; an additional 14 PAHs were not tested in the same study as 

4 benzo[a]pyrene. The remaining 28 PAHs either have only cancer-related endpoint data, or have 

5 neither bioassays nor cancer-related endpoint data. Bioassays without benzo[a]pyrene were 

6 considered in the weight of evidence evaluation for individual PAHs (Section 6.1). Studies that 

7 provided only information on PAH mixtures or PAH metabolites were not reviewed or 

8 summarized for this analysis. 

9 References in the database were sorted by keyword into the following major categories: 

10 cancer bioassays, in vivo studies of cancer-related endpoints, and in vitro studies of cancer11 related endpoints. These categories were further divided by route (for bioassays) or by endpoint 

12 (for cancer-related endpoints). Each study was reviewed, and critical study details were 

13 extracted into tables (Tables 4-1 through 4-14) for each individual endpoint. Studies with data 

14 on selected PAHs and benzo[a]pyrene were used, even if a particular PAH has not been 

15 evaluated by U.S. EPA or IARC for carcinogenicity. Studies were included in the analysis if the 

16 following selection criteria were met: 

17 

18 • Benzo[a]pyrene was tested simultaneously with another PAH; 

19 

20 • A statistically increased incidence of tumors was observed with benzo[a]pyrene 

21 administration; 

22 

23 • Benzo[a]pyrene produced a statistically significant change in a cancer-related endpoint 

24 finding; 

25 

26 • Quantitative results were presented; 

27 

28 • The carcinogenic response observed in either the benzo[a]pyrene- or other PAH-treated 

29 animals at the lowest dose level was not saturated (i.e., tumor incidence at the lowest 

30 dose was <90%); and 

31 

32 • There were no study quality concerns or potential confounding factors that precluded use 

33 (e.g., no concurrent control, different vehicles, strains, etc. were used for the tested PAH 

34 and benzo[a]pyrene; use of cocarcinogenic vehicle; PAHs of questionable purity; 

35 unexplained mortality in treated or control animals). 

36 
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Table 4-1. Study summaries: dermal bioassays of benzo[a]pyrene and at least one other PAH 

Record 

number Reference 

Mouse

a

strain Exposure Follow up Vehicle Promoter Tumor type

Positive 

result Negative result 

Meets selection 

criteria? Comments 

Complete carcinogenicity studies

480 Bingham and Falk, 

1969 

CH3/He 3 times/wk 50 wk Toluene 

or n-dodecane 

None Malignant 

and benign 

BaA No BaP administered in different vehi

n-Dodecane cocarcinogenic with B

No concurrent untreated, toluene o

dodecane control. 

600 Habs et al., 1980 NMRI 2 times/wk 

(4 times for CO) for life 

until moribund or dead 

Acetone (DMSO for CO) 

None Papilloma, 

carcinoma, sarcoma 

BbF BkF, BjF, CPcdP, 

CO, IP 

Yes 

22390 Wynder and 

Hoffmann, 1959a 

Swiss 3 times/wk 6–14 mo cyclohexane None Papilloma, carcinoma BbF, BjF BghiF, BkF No Deaths prior to first tumor appeara

No concurrent control. 

19320 LaVoie et al., 1979 HA/ICR 

Swiss albino 

3 times/wk Unspecified Acetone None Unspecified CH, BbF, 

BjF, DBaeP, DBahP, DBaiP 

AC, Pyr, BghiF, BkF, AA, BeP, DBelP, IP, BghiP, N23eP 

No Reiterates data published elsewher

22400 Wynder and 

Hoffmann, 1959b 

Swiss 3 times/wk 10–22 mo Acetone None Papilloma, 

carcinoma 

CH, DBahA, DBaiP 

AC, BeP, Pyr, FA No Deaths prior to first tumor appeara

Not clear if BaP administered 

simultaneously. No concurrent control. 

13640 Cavalieri et al., 1983 Swiss 2 times/wk 

for 48 wk 

Until 2 cm tumor or 61 wk 

Acetone None Papilloma, 

adenoma, carcinoma 

CPcdP Yes Reports both incidence and 

multiplicity. 

13650 Cavalieri et al., 

1981b 

Swiss 2 times/wk 

for 30 wk 

Until 2 cm tumor, moribund or 57 wk 

Acetone None Primarily 

squamous cell carcinoma 

CPcdP ACEP Yes Tumor incidence not useable becau

BaP tumor incidence was 100%. Tumor multiplicity data available dose-response assessment. 

620 Hoffmann and 

Wynder, 1966 

Ha/ICR/

Mil Swiss 

3 times/wk for 12 mo 

Up to 15 mo Dioxane None Papillomas DBaeP, 

DBahP, DBaiP, DBaeF 

 Yes Paper in German. Paper reports 

compound as DBalP; LaCassagne al. (1968) state that it is actually DBaeF. DBahP incidence ≥ 90% alowest dose. 

17660 Cavalieri et al., 1977 Swiss 2 times/wk 

for 30 wk 

Until moribund, dead, or after 70 wk 

Acetone None Papilloma, 

keratoacanthoma, carcinoma 

DBahP, AA 

BaA Yes DBahP incidence ≥ 90% at lowest 

dose. 

610 Higginbotham et al., 

1993 

Swiss 2 times/wk 40 wk Acetone None Papilloma, 

carcinoma 

DBalP No No tumors with BaP. 

19760 Masuda and 

Kagawa, 1972 

Ha/ICR/

Mil Swiss 

3 times/wk for 60 applications 

7 months Dioxane None Unspecified DBalP No No concurrent untreated or vehicle 

control; lowest dose DBalP gave 100% incidence. 

18570 Hecht et al., 1974 Ha/ICR/

Mil Swiss 

3 times/wk for 17 mo 

72 wk Acetone None Unspecified CH No BaP dose not reported. 
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Table 4-1. Study summaries: dermal bioassays of benzo[a]pyrene and at least one other PAH 

Record 

number Reference 

Mouse

a

strain Exposure Follow up Vehicle Promoter Tumor type

Positive 

result Negative result 

Meets selection 

criteria? Comments 

21310 Shubik et al., 1960 Syrian 

golden hamster 

2 times/wk for 10 wk 

75 wk Mineral 

oil 

None None DBahA, BaA No Small number of animals (5/sex/do

23310 Pfeiffer and Allen, 

1948 

Rhesus monkey 

various Various Sesame 

oil 

None Various Multiple No Sequential exposure to multiple 

compounds; no concurrent untreatcontrol. 

23840 Barry et al., 1935 Unspecified 2 times/wk 1–2+ yr Benzene None Epithelioma, papilloma Multiple No Test compounds from various sour

gave differing results; purity may bsuspect; use of benzene vehicle confounds tumorigenicity results; 

benzene or untreated control. 

Initiation studies

24800 Nesnow et al., 1984 SENCAR Single 31 wk Acetone 12-O-tetradecanoylphorbol-13-

acetate (TPA) 2 μg 2 times/wk for 30 wk 

Papilloma BeAC, 

BlAC 

 Yes Reports both incidence and 

multiplicity. 

21410 Slaga et al., 1978 CD-1 Single 27 wk Acetone TPA 10 μg 

2 times/wk for 26 wk 

Papilloma BaA Yes Tumor incidence data not useable

because BaP gave 93% tumor 

incidence. Tumor multiplicity datavailable for dose-response assessment. 

630 LaVoie et al., 1982 Crl:CD1[ICR]

BR 

10 subdoses every other d

Unspecified Acetone TPA 3 

times/wk 20 wk 

Primarily squamous cell 

papilloma 

BbF, BjF, BkF 

 Yes Reports both incidence and 

multiplicity. 

16310 Weyand et al., 1992 Crl:CD-1 5 or 

10 applications given every other d

Until 

promotion 

complete 

Acetone TPA 2.5 μg 

3 times/wk for 20 wk 

Unspecified BjF Yes Tumor incidence data not useable 

because BaP gave 100% tumor 

incidence. Tumor multiplicity datavailable for dose-response assessment. DNA adducts, mutagenicity also evaluated. 

10200 El-Bayoumy et al., 

1982 

Crl:CD1[ICR]BR 

10 subdoses every other d

Unspecified Acetone TPA 2.5 μg 

3 times/wk for 25 wk 

Primarily squamous cell 

papilloma 

CH Pery, Pyr Yes Tumor incidence data not useable 

because single dose CH gave 100% 

tumor incidence; BaP gave 90% tuincidence. Tumor multiplicity datavailable for dose-response assessment. 

18570 Hecht et al., 1974 Ha/ICR/

Mil Swiss 

10 subdoses every other d

Until 

promotion 

complete 

Acetone TPA 2.5 μg 

3 times/wk for 20 wk 

Unspecified CH Yes Reports both incidence and 

multiplicity. 
66 DRAFT – DO NOT CITE OR QUO

Table 4-1. Study summaries: dermal bioassays of benzo[a]pyrene and at least one other PAH 

Record 

number Reference 

Mouse

a

strain Exposure Follow up Vehicle Promoter Tumor type

Positive 

result Negative result 

Meets selection 

criteria? Comments 

22500 Van Duuren et al., 

1966 

ICR/HA Single 63 wk Acetone Croton resin, 

25 μg 3 times/wk 

Papilloma, carcinoma 

CH, BbF BghiF No BaP gave 100% tumor incidence. 

Corollary data with acetone only a

promotion agent not included. 

24300 Rice et al., 1985 CD-1 10 subdoses 

every other d

Until 

promotion 

complete 

Acetone TPA 

0.0025% 3 times/wk for 20 wk 

Unspecified CH, 

CPdefC 

 Yes Tumor incidence data not useable 

because all compounds gave >90% 

tumor incidence. Tumor multiplicdata available for dose-response assessment. 

19320 LaVoie et al., 1979 HA/ICR 

Swiss albino 

10 subdoses every other d

Until 

promotion 

complete 

Acetone or dioxane 

TPA 2.5 μg 3 times/wk for 20 wk or croton oil 2.5% 3 times/wk 

Unspecified CH, 

DBaeP, DBahP, DBaiP, 

N23eP 

FA, AA, DBelP, BghiP, IP 

No Reiterates data published elsewher

21420 Slaga, et al., 1980 Sencar Single 15 wk Acetone TPA 2 μg 

2 times/wk 

Papilloma CH, 

DBahA, 

BeP, DBacA Yes Not clear if BaP done simultaneou

but protocol, vehicle, follow up sa

Reports both incidence and multiplicity. 

15640 Raveh et al., 1982 Sencar Single 25 wk Unspecified TPA 2 μg 2 times/wk 

for 25 wk 

Papilloma CPcdP Yes Reports both incidence and 

multiplicity. 

620 Hoffmann and 

Wynder, 1966 

Ha/ICR/

Mil Swiss 

Single 6 mo Dioxane Croton oil Papillomas DBaeF, 

DBaeP, DBahP, DBaiP, 

N23eP 

IP, AA, BghiP, DBelP 

Yes Paper reports compound as DBalP

LaCassagne et al. (1968) state that actually DBaeF. 

610 Higginbotham et al., 

1993 

Sencar Single 27 wk Acetone TPA 

2.6 nmol, 2 times/wk 

Papillomas, few carcinomas 

DBalP No No tumors with BaP. 

13660 Cavalieri et al., 1991 Sencar Single 16 wk and 

27 wk (two experiments)

Acetone TPA 

3.24 nmol 2 times/wk for 11 wk 

Primarily 

papilloma 

DBalP Yes Tumor incidence data not useable 

because lowest dose DBalP gave 

>90% tumor incidence. Tumor multiplicity data from both experiments available for doseresponse assessment. 

19360 LaVoie et al., 1985 Crl:CD/1 

(ICR)BR 

10 subdoses every other d

Unspecified Acetone TPA 2.5 μg 

3 times/wk for 20 wk 

Unspecified AC Yes 

13650 Cavalieri et al., 

1981b 

CD-1 10 subdoses 

every other d

57 wk Acetone TPA 

0.017 μmol 2 times/wk for 40 wk 

Papilloma CPcdP ACEP Yes Reports both incidence and

multiplicity. 
67 DRAFT – DO NOT CITE OR QUO

Table 4-1. Study summaries: dermal bioassays of benzo[a]pyrene and at least one other PAH 

Record 

number Reference 

Mouse

a

strain Exposure Follow up Vehicle Promoter Tumor type

Positive 

result Negative result 

Meets selection 

criteria? Comments 

20830 Roe, 1962 Albino Single Until 

promotion 

complete 

Acetone Croton oil 

once/wk for 20 wk 

Papilloma PH No BaP not simultaneous. 

16440 Wood et al., 1980 CD-1 Single 27 wk Acetone TPA 16 nmol 

2 times/wk for 26 weeks 

Unspecified Pyr, CPcdP Yes 

17450 Brune et al., 1978 NMRI Unspecified Unspecified Unspecified TPA Unspecified AC No Study design not reported. Results reported qualitatively. 18680 Hoffmann et al., 

1972 

Ha/ICR/

Mil Swiss 

10 subdoses every other d

Until 

promotion 

complete 

Acetone Croton oil 

2.5% for 20 wk 

Unspecified FA Yes 

19420 LaVoie et al., 1981 HA/ICR 

Swiss albino 

10 subdoses every other d

Unspecified Acetone TPA 2.5 μg 

3 times/wk for 20 wk 

Unspecified PH Yes 

13660 Cavalieri et al., 1991 SENCAR Single 27 wk Acetone None Primarily 

papilloma 

DBalP Yes Initiating dose only; no promoter. 

Tumor incidence data not useable 

because lowest dose DBalP gave 

>90% tumor incidence. Tumor multiplicity data available for doseresponse assessment. 

15700 Rice et al., 1988 CD-1 10 subdoses 

every other d

24 wk Acetone TPA 2.5 μg 

3 times/wk for 20 wk 

Unspecified CH, 

BbcAC, CPdefC 

 Yes Not clear if BaP done simultaneou

for all PAHs. 

Cocarcinogenicity studies

18700 Horton and 

Christian, 1974 

C3H 2 times/wk 

for 80 wk 

82 wk n-Dodecane/

decalin mixture 

None Carcinoma, 

papilloma 

DBacA, Pyr 

CH, FA, Tphen, Pery, 

No Not clear if BaP done simultaneou

Experiments with decalin (noncarcinogen) and 50/50 decalindodecane mix (cocarcinogenic). Ndata for BaP in 50/50 mix. No vehcontrol in decalin. 

21430 Slaga et al., 1979 CD-1 Single 30 wk Acetone TPA 10 μg 

2 times/wk for 30 wk 

Papilloma BeP No No concurrent control. Study aime

exploring interactions; not clear if done simultaneously. 

21840 Van Duuren and 

Goldschmidt, 1976 

ICR/Ha Swiss 

3 times/wk 368 or 440 d Acetone None Papilloma Pyr, BghiP, BeP, 

FA 

Yes 

21850 Van Duuren et al., 

1973 

ICR/HA 3 times/wk 

for 52 wk 

52 wk Acetone None None Pyr, BghiP, BeP No Qualitative results reported. 

21920 Warshawsky et al., 

1993 

C3H/HEJ 2 times/wk Until lesion 

developed or 104 wk 

Toluene or n-dodecane 

None Unspecified AC, CH, Pyr, FA, 

PH 

No No tumors with BaP. 

aExcept where noted, all studies were conducted in mice. 
68 DRAFT – DO NOT CITE OR QUO

1 

Table 4-2. Study summaries: intraperitoneal bioassays of benzo[a]pyrene and at least one other PAH 

Record 

number Reference Mouse strain

a Exposure 

Follow 

up Vehicle

Target 

organ(s) Tumor type(s) 

Positive 

result 

Negative 

result 

Meets selection 

criteria? Comments 

Newborn mouse studies

13610 Busby et al., 

1984 

SwissWebster BLU:Ha (ICR) 

1st, 8th, 15th d 26 wk DMSO Lung Adenoma, 

adenocarcinoma 

FA Yes Tumor incidence data not useable 

because lowest dose BaP gave >90% 

tumor incidence. Tumor multiplicity data available for dose-response assessment. 

17560 Busby et al., 

1989 

SwissWebster BLU:Ha (ICR) 

1st, 8th, 15th d 26 wk DMSO Lung Adenoma, 

adenocarcinoma 

FA Pyr, CH Yes Reports both incidence and multiplicit

640 LaVoie et al., 

1987 

CD-1 1st, 8th, 15th d 52 wk DMSO Lung, 

liver 

Adenoma, hepatoma 

BbF, BjF BkF, IP Yes 

7510 LaVoie et al., 

1994 

CD-1 1st, 8th, 15th d 12 mo DMSO Lung, 

liver 

Foci, adenoma, carcinoma 

FA Yes Reports both incidence and multiplicit

22040 Weyand and 

LaVoie, 1988 

CD-1 1st, 8th, 15th d Not 

reported 

DMSO Lung, 

liver 

Unspecified Not 

reported 

 No Abstract only; dose-response informat

not included. 

22510 Wislocki et al., 

1986 

CD-1 1st, 8th, 15th d 12 mo DMSO Lung, 

liver, lymphatic system 

Adenoma, carcinoma, lymphoma 

CH, BaA Pyr Yes Reports both incidence and multiplicit

Studies in adult A/J mice

11190 Mass et al., 1993 A/J Single 8 mo Tricaprylin Lung Adenoma, carcinoma BjAC No Reiterates data reported elsewhere (Record 24590). 23960 and 23450 

Nesnow et al., 

1998a, 1995 

A/J Single 8 mo Tricaprylin Lung Adenoma BbF, DBahA, 

CPcdP 

 No Reiterates data reported elsewhere 

(Record 24590). 

22670 Nesnow et al., 

1996 

A/J Single 8 mo Tricaprylin Lung Adenoma BbF, DBahA, 

CPcdP 

 No (Reiterates data reported elsewhere 

(Record 24590).) 

24590 Nesnow et al., 

1998b 

A/J Single 8 mo Tricaprylin Lung Adenoma CPcdP, BbF, 

DBahA, BjAC, DBalP 

 Yes Raw data obtained courtesy of S. 

Nesnow. Tumor incidence for BaP w

100% at lowest dose with significant increase over control; tumor multiplicdata available. 

20920 Ross et al., 1995 A/J Single 240 d Tricaprylin Lung Adenoma BbF, DBahA, 

CPcdP 

Pyr No Reiterates data reported elsewhere 

(Record 24590). 

aAll studies were conducted in mice. 
69 DRAFT – DO NOT CITE OR QUO

1 

Table 4-3. Study summaries: subcutaneous bioassays of benzo[a]pyrene and at least one other PAH 

Record 

number Reference Species Strain 

Exposure 

site Exposure 

Follow 

up Vehicle 

Target 

organ(s) 

Tumor 

type(s) 

Positive 

result 

Negative 

result 

Meets selection 

criteria? Comments 

23840 Barry et al., 1935 Mouse Unspecified Unspecified Single 1–2+ yr Lard Injection site Sarcoma Multiple No Test compounds fro

various sources gavdiffering results; pumay be suspect; no untreated control. 

220 Bryan and 

Shimkin, 1943 

Mouse C3H Right axilla Single until 

20 mm tumor 

Tricaprylin Injection 

site 

Unspecified DBahA No No concurrent untre

control. 

18350 Grant and Roe, 

1963 

Mouse Albino Neck 1st d after 

birth 

52–62 wk Aqueous 

gelatin 

Lung Adenoma PH Yes 

23200 Homburger et al., 

1972 

Hamster Various Groin Single 52 wk Tricaprylin Injection 

site; lung 

Various BaA No Study aimed at 

evaluating strain specificity of tumorigenicity. BaA results equivocal. Nclear if BaP treatmesimultaneous. "Agemice used as controaged mice allowed tlive 16 weeks longe

660 Pfeiffer, 1977 Mouse NMRI Neck Single 114 wk Tricaprylin Injection 

site 

Sarcoma DBahA No Less than 10% of 

100 control mice aliat 114 wk; control dnot provided. 

23310 Pfeiffer and Allen, 

1948 

Monkey Rhesus Various Various variable Sesame oil Various Various Multiple No Sequential exposure 

multiple compoundsconcurrent untreated control. 

24290 Rask-Nielson, 

1950 

Mouse Street Thymus, 

lung, mammary area 

Single 30 mo Paraffin Various Various DBahA No Number of control a

exposed varies by tumor type reportedBaP nontumorigenicDBahA results equivocal; results unclear. 

24310 Roe and Waters, 

1967 

Mouse Swiss 

albino 

Not 

specified 

1st d after 

birth 

50–60 wk Aqueous 

gelatin 

Liver Hepatoma PH No Study methodology 

results not detailed; results equivocal. 

21560 Steiner, 1955 Mouse C57BL Interscapular Single 22–28 mo Tricaprylin Injection 

site 

Sarcoma DBahA, 

BaA, CH 

AC, PH No No concurrent untre

control; study aimed evaluating interactio

2 
70 DRAFT – DO NOT CITE OR QUO

1 

Table 4-4. Study summaries: oral bioassays of benzo[a]pyrene and at least one other PAH 

Record 

number Reference Species Strain 

Exposure 

route Exposure Follow up

Target 

organ(s) 

Tumor 

type(s) 

Positive 

result 

Negative 

result 

Meets selection 

criteria? Comments 

17280 Biancifiori and Caschera, 

1962 

Mouse BALB/c Gavage 2 times/wk, 

15 wk 

Variable; 50–60 wk 

Mammary gland 

Carcinomas and sarcomas 

DBahA No Tumors observed after DBahA on

in pseudopregnant mice, not virgimice. 

23880 Huggins and Yang, 1962 Rat SpragueDawley Gavage Single Not reported Mammary gland Unspecified BaA, PH No Untreated control information not included. 

2 
71 DRAFT – DO NOT CITE OR QUO

1 

Table 4-5. Study summaries: other route bioassays of benzo[a]pyrene and at least one other PAH 

Record 

number Reference Species Strain Exposure route Exposure Follow up Vehicle 

Target 

organ(s) Tumor type(s)

Positive 

result 

Negative 

result 

Meets 

selection 

criteria? Commen

21750 Topping et al., 1981 Rat F344 Implantation in

transplanted tracheas 

Release from pellet 

28 mo Beeswax 

pellet 

Tracheal epithelium 

Carcinoma, sarcoma 

 BeP No Interaction 

information included. 

17620 Cavalieri et al., 

1988b 

Rat SpragueDawley Intramammillary Single 20 wk None Mammary Adeno- carcinoma, 

adenofibroma, fibrosarcoma 

 DBahA, 

BaA 

No Control data 

from untreamammary glands of sarats. 

13660 Cavalieri et al., 1991 Rat SpragueDawley Intramammillary Single Until 2 cm tumor or 

24 wk 

Trioctanoin Mammary, 

other 

Adenocarcinoma, adenofibroma, fibrosarcoma, squamous cell carcinoma 

DBalP No DBalP 

produced 

tumors in alanimals at tlowest dose

21620 Sugiyama, 1973 Rat Long Evans Intramuscular Single 9 mo Sesame oil Injection 

site 

Sarcoma BaA No BaP gave 10

tumor incidence. 

20280 Pataki and Huggins, 

1969 

Rat SpragueDawley Intravenous 3 doses 3 d apart 98 d Lipid emulsion Mammary Unspecified BaA No No control group. 

17940 Deutsch-Wenzel et 

al., 1983 

Rat OsborneMendel Lung implantation Release from pellet Until moribund 

or dead 

Beeswax/ trioctanoin 

Lung Carcinoma, 

sarcoma 

BbF, BjF, BkF, IP, AA, BghiP 

BeP Yes 

22000 Wenzel-Hartung et 

al., 1990 

Rat OsborneMendel Lung implantation Release from pellet Until moribund 

or dead 

Beeswax/ trioctanoin 

Lung Carcinoma CH, 

DBahA

PH Yes 

21500 Solt et al., 1987 Hamster Syrian 

golden 

Painting buccal 

pouch 

2 times/wk for 20 wk 

Up to 44 wk 

Paraffin oil Buccal 

pouch 

Carcinoma BaA No Fewer than 

animals per group; negaresult. 

23910 Nikonova, 1977 Mouse A Subcutaneous (F0) 

and transplacental (F1) 

GD 18 or 19 

1 yr Sunflower 

oil 

Lung, mammary, liver, injection site 

Adenoma Pyr No Transplacen

exposure noquantified. 

2 
72 DRAFT – DO NOT CITE OR QUO

1 

Table 4-6. Study summaries: in vivo DNA adducts with benzo[a]pyrene and at least one other PAH 

Record 

number Reference 

Route of 

administration 

Exposure 

frequency 

Hours between 

dosing and 

sacrifice Tissue analyzed

Method of 

analysis PAHs evaluated

a

Meets selection 

criteria? Comments 

6210 Arif et al., 1997 Intramammillary Single dose 48 Mammary 

epithelium, lung 

[32P] postlabeling DBalP Yes 

17420 Brookes and 

Lawley, 1964 

Dermal Single dose various to ~12 d Skin [

3H]- prelabeling DBacA, DBahA No Data on individual compounds no

reported. 

17630 Cavalieri et al., 

1981a 

Dermal Single dose 4, 24 Skin [

3H] or [14C] 

prelabeling 

CPcdP, ACEP Yes 

18810 Hughes and 

Phillips, 1990 

Dermal Single dose 0.5, 1, 2, 4, 7, 21, 

84 d 

Skin, lung [32P] postlabeling DBalP, DBaeP, 

DBahP, DBaiP 

Yes 24-hr experiment with DBaeP and 

DBalP; 84-d experiment with all. 

18790 Hughes and 

Phillips, 1991 

Dermal Single dose 24 Skin [32P] postlabeling DBaeP No No quantitative information; abstr

only. 

10900 Koganti et al., 

2000 

Oral-diet 14 d not stated Lung [32P] postlabeling BcFE, BaFE, BbFE No Not quantified. 

13200 Li et al., 2002 Oral-gavage or 

oral-diet 

1 time/d for 1–4 d; diet 14 d 

 Mammary gland 

and liver; lung 

[32P] postlabeling BcFE No Not quantified; BaP administered 

gavage, BcFE admin in diet. 

11190 Mass et al., 

1993 

Intraperitoneal Single dose 24, 48, 72 Lung [32P] postlabeling BjAC Yes 

8010 Nesnow et al., 

1993b 

Intraperitoneal Single dose 1, 3, 7, 14, 28, 56 d Lung, liver, 

peripheral blood 

lymphocytes 

[32P] postlabeling BbF Yes Peaks differ temporally; study als

correlates number of adducts in organs. 

22670 Nesnow et al., 

1996 

Intraperitoneal Single dose 7 d Lung [32P] postlabeling BbF, DBahA, 

CPcdP 

No Not quantified. 

23960 Nesnow et al., 

1995 

Intraperitoneal Single dose 7 d Lung [32P] postlabeling BbF, DBahA, 

CPcdP 

No Not quantified. 

24590 Nesnow et al., 

1998a 

Intraperitoneal Single dose various to 21 d Lung [32P] postlabeling BbF, CPcdP, 

DBahA, DBalP 

Yes Used data from Ross et al., 1995 (

20920) to calculate slope. 

22810 Phillips et al., 

1979 

Dermal Single dose 19, 24, 48, 72, 96, 

120, 144 

Skin [

3H]-Prelabeling BaA, DBacA, 

DBahA 

Yes 

20650 Reddy et al., 

1984 

Dermal 4 doses (0, 6, 

30, 54 hr 

24 Skin [32P] postlabeling AC, BaA, BghiP, 

BeP, CH, DBacA, DBahA, Pery, Pyr 

No Semiquantitative data only. 

20920 Ross et al., 

1995 

Intraperitoneal Single dose 0, 1, 3, 5, 7, 14, 21 d Lung [32P] postlabeling BbF, CPcdP, 

DBahA 

No Reiterates data published elsewhe

(see 24590) 

16310 Weyand et al., 

1992 

Dermal Single dose 24 Skin [32P] postlabeling BjF No Not quantified. 

22040 Weyand and 

LaVoie, 1988 

Intraperitoneal Postnatal d 1, 

8, 15 

24 Lung, liver [32P] postlabeling BbF, BjF, BkF No No quantitative data; abstract only
73 DRAFT – DO NOT CITE OR QUO

Table 4-6. Study summaries: in vivo DNA adducts with benzo[a]pyrene and at least one other PAH 

Record 

number Reference 

Route of 

administration 

Exposure 

frequency 

Hours between 

dosing and 

sacrifice Tissue analyzed

Method of 

analysis PAHs evaluated

a

Meets selection 

criteria? Comments 

24790 Kligerman et 

al., 2002 

Intraperitoneal and oral 

Single dose 7 d Peripheral blood 

lymphocytes 

[32P] postlabeling BaA, BbF, CH Yes Data in both rats and mice. 

aPositive findings were reported for all PAHs evaluated. 
74 DRAFT – DO NOT CITE OR QUO

1 

Table 4-7. Study summaries: in vivo clastogenicity or sister chromatid exchange with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Species Strain 

Route of 

administration Vehicle Exposure

Hours between 

dosing and 

sacrifice 

Tissue 

analyzed 

Clastogenic 

endpoint

Positive 

results 

Negative 

results 

Meets 

selection 

criteria? Comments 

24740 Allen et al., 1999 Mice A/J or 

p53 +/+, 

+/-, and -/- 

Intraperitoneal Tricaprylin Single 48 or 72 hr Bone

marrow or 

peripheral blood 

Micronuclei DBalP Yes 

14270 He and Baker, 

1991 

Mice HRA/Skh 

hairless 

Dermal Acetone Single 24 hr Keratinocytes Micro- nuclei CH Pyr Yes 

17190 Bayer, 1978 Hamsters Chinese Intraperitoneal Tricaprylin Single 24 hr for 

aberrations; 30 hr for micronuclei 

Bone marrow 

Gaps, 

breaks, 

micronuclei, SCEs 

PH (high dose only) 

 Yes 

19030 Katz et al., 1981 Mice B6C3F1/

BR 

Intraperitoneal DMSO At 0 and 

24 hr 

various; 24, 30, 48, 72 hr after last dose 

Bone marrow 

micronuclei DBaiP, AC, 

BghiP, Pyr

No No quantitative da

24720 Kligerman et al., 

1986 

Mice C57BL6 Gavage Corn oil Single 23.5–25 hr Peripheral 

blood 

SCEs BlAC Yes 

24790 Kligerman et al., 

2002 

Mice and rats 

CD-1 Swiss mice; CD rats 

Oral and intraperitoneal 

Sunflower seed oil 

Single 7 d Whole blood 

or mononuclear leukocytes 

SCE, micronuclei 

BaA, BbF, CH 

 Yes All positive for SC

via intraperitoneal administration; mixed results for oadministration. 

20200 Oshiro et al., 

1992 

Mice CD-1 Peroral PEG 1 time/d, 

4 d 

24 hr after 2nd and 4th treatment 

Peripheral 

blood 

Micronuclei Pyr, AC No No quantitative da

published as abstr

20230 Paika et al., 1981 Mice CBA/J Intraperitoneal DMSO single 16–20 hr Bone 

marrow 

SCEs Pyr No No quantitative da

20950 RoszinskyKocher et al., 

1979 

Hamsters Chinese Intraperitoneal Tricapryline 2 doses 

24 hr apart 

24 hr after 2nd treatment 

Bone marrow 

SCEs, aberrations 

PH, CH, DBahA, BaA, BbF, BeP

AC Yes Positive results fo

SCEs, not aberrations. 

21050 Salamone et al., 

1981 

Mice B6C3F1 Intraperitoneal Not 

specified 

2 doses 24 hr apart 

24, 48, 72 hr after 2nd treatment 

Bone marrow 

Micronuclei AC, Pyr Yes 

21770 Tsuchimoto and 

Matter, 1981 

Mice CD-1 Intraperitoneal DMSO 2 doses 

24 hr apart 

6 hr after 2nd treatment 

Bone marrow 

Micronuclei Pyr Yes 

21390 Sirianni and 

Huang, 1978 

Mice C3H/St V79 cells in diffusion chamber 

implanted in 

peritoneal 

cavity of mice 

 Chinese 

hamster V79 cells 

SCEs AC, Pyr, 

Pery 

Yes 

21620 Sugiyama, 1973 Rats LongEvans Intravenous Lipid emulsion Single 12, 24 hr Bone marrow Gaps, breaks BaA Yes 
75 DRAFT – DO NOT CITE OR QUO

1 

Table 4-8. Study summaries: in vivo mutagenicity with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Species/strain 

Route of 

administration 

Exposure 

frequency/follow up Mutagenic endpoint 

Positive 

result 

Negative 

result 

Meets 

selection criteria? Comments 

18130 Fahmy and Fahmy, 

1980 

Drosophila 

melanogaster 

Suspension in media 

48–72 hr Somatic mutation; eye color 

mosaicism 

 BaA Yes 

13980 Frolich and Wurgler, 

1990 

D. melanogaster Suspension in 

media 

48–72 hr Somatic mutation and 

recombination test (SMART); wing spots 

 BaA No Inconsistent results for BaA; signific

effects only seen with cross-breedinstrains selected for enhanced metaboactivity (not standard strains). 

11190 Mass et al., 1993 A/J mice Intraperitoneal 3 d/8 mo Mutations in codon 12 of 

the Ki-ras oncogene; 

polymerase chain reaction 

(PCR) and DNA sequencing of lung tumor DNA 

BjAC No Quantitative dose-response data wer

not available. Different mutation sequences observed; GGT→TGT foBaP and GGT→CGT for BjAC; mutation sequence for BjAC may correlate with cyclopenta-adduct formation. 

23960 Nesnow et al., 1995 A/J mice Intraperitoneal Single injection/ 

8 mo 

Mutations in codon 12 of the Ki-ras oncogene; PCR and DNA sequencing of lung tumor DNA 

BbF, DBahA, CPcdP 

 No Quantitative dose-response data wer

not available. GGT→TGT mutationfor BaP and BbF; GGT→CGT for CPcdP; no mutations seen for DBah

22670 Nesnow et al., 1996 A/J mice Intraperitoneal Single injection/ 

8 mo 

Mutations in codon 12 of the Ki-ras oncogene; PCR and DNA sequencing of lung tumor DNA 

BbF, DBahA, CPcdP 

 No Quantitative dose response data wer

available. GGT→TGT mutations foBaP and BbF; GGT→CGT for CPcdno mutations seen for DBahA. 

24590 Nesnow et al., 1998b A/J mice Intraperitoneal Single injection/ 

8 mo 

Mutations in codons 12 and 61 of the Ki-ras oncogene; PCR and DNA sequencing of lung tumor DNA 

BbF, DBahA, CPcdP, BjAC, DBalP 

 No Quantitative dose-response data wer

not available. Mutations in codon 1GGT→TGT for BaP, BbF, and DBaGGT→CGT for CPcdP and BjAC; nmutations seen for DBahA; GTT mutations seen for all other PAHs. ODBalP caused mutations in codon 6

21370 Simmon et al., 1979 Swiss Webster 

mice 

PAHs intramuscular or 

peroral; 

microorganisms intraperitoneal 

Single injection/4 hr Intraperitoneal host 

mediated assay; mutagenicity in S. typhimurium and S. cerevisiae of recovered microorganisms 

 AC, BaA, 

BeP, CH, PH 

No Assay was not considered sensitive 

enough for detecting carcinogens. 

21830 Valencia and 

Houtchens, 1981 

D. melanogaster Filter feeding 48–72 hr Sex-linked recessive lethal

test 

 Pyr No Results were negative for BaP. 

22450 Zijlstra and Vogel, 

1984 

D. melanogaster Abdominal 

injection 

Not applicable Sex-linked recessive lethal 

test; 2–3 translocation and ring-X loss 

 BaA No Results were negative for BaP. 
76 DRAFT – DO NOT CITE OR QUO

1 

Table 4-9. Study summaries: in vitro bacterial mutagenicity with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Salmonella strain(s) Activation system Positive result Negative result

Meets 

selection 

criteria? Comments 

17030 Andrews et al., 

1978 

TA100, TA1527, TA1538 

Ar S9 and others AA, DBahA, DBajA, DBacA, BghiP, 

BeP 

 Yes TA100 results include BaP. 

23830 Baker et al., 1980 TA100 Guinea pig MC S9 

and others 

DBaiP, BaA, DBacA, DBahA Yes 

23660 Bartsch et al., 1980 TA100, TA1535, 

TA98 

Rat MC S9 BaA Yes 

17380 Bos et al., 1988 TA98, TA100 Rat Ar S9 PH, Pyr Yes Qualitative data for other PAHs (no BaP

quantitative data with BaP comparison fPH and Pyr in TA100. 

9560 Carver et al., 1985 TA98, TA100 S9 Pery No The response varied at different 

concentrations of S9; BaP was more potat low S9 while Pery was more potent at high S9. 

17590 Carver et al., 1986 TA100 Ar rat and Ar hamster 

S9 

BaA, BghiF, Pery Yes Qualitative data also presented for other 

PAHs. S9 concentration varied; 400 μL/plate optimal. 

17630 Cavalieri et al., 

1981a 

TM677 Ar S9 CPcdP, ACEP, Pyr Yes BaP data from previous publication used

Dose-response data not provided for Pyr

9620 Chang et al., 2002 TA100 Rat Ar S9 BghiF, BcPH Yes 24030 De Flora et al., 1984 TA1535, TA1537, 

TA1538, TA98, TA100 

Rat AR S9 BaA, Pery, BeP AC Yes 

13860 Devanesan et al., 

1990 

TA100, TA98 Rat Ar S9 DBaeP, DBalP No No concurrent control. 

18030 Dunkel et al., 1984 TA1535, TA1537, 

TA1538, TA98, TA100 

Rat, mouse, hamster Ar S9 

BaA, BeP, PH, Pyr AC No Dose-response data not provided. 

18050 Eisenstadt and 

Gold, 1978 

TA1537, TA100 Rat Ar S9 CPcdP Yes 

18180 Florin et al., 1980 TA98, TA100 Rat Ar and MC S9 BaA, CH, Pery, CO Yes 24080 Gibson et al., 1978 TA1535, TA1537, 

TA1538, TA98 

Nonenzymatic 

(gamma radiation) 

BaA, BghiP, CH, FE, Pyr DBahA, AC, 

Pic, Tphen 

Yes AN, PH also tested; toxicity interfered w

mutagenicity testing. 

14080 Gold and 

Eisenstadt, 1980 

TA100 Rat MC S9 CPcdP Yes BaP and CPcdP maximal responses 

occurred at different S9 levels. 

14170 Guthrie et al., 1982 TA98, TA100 Rat Ar S9 compare to 

PGS from ram seminal vesicles 

BaA, CH No BaP tested in TA98, BaA and CH tested 

TA100. 

14260 Hass et al., 1981 TA98, TA100 Rat Ar S9 BeP Yes 
77 DRAFT – DO NOT CITE OR QUO

Table 4-9. Study summaries: in vitro bacterial mutagenicity with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Salmonella strain(s) Activation system Positive result Negative result

Meets 

selection 

criteria? Comments 

18650 Hermann, 1981 TA98 Rat Ar S9 BbA, BaA, CH, FA, Tphen, BeP, 

DBacA, DBahA, BbF, Pery, DBalP, DBaiP, AA, CO 

AC, PH, FE, Pyr, BbFE 

Yes 

10670 Johnsen et al., 1997 TA98 Rat control or PB S9 BjAC, BlAC Yes 19000 Kaden et al., 1979 TM677 Rat Ar or PB S9 AN, ANL, Pyr, BbFE, CPcdP, BaA, CH, 

Tphen, FA, BeP, Pery, BghiP, AA, DBacA, DBahA, DBbeF 

FE, AC, PH, Pic, CO 

Yes Mutagenic activity relative to BaP repor

24680 Lafleur et al., 1993 TM677 Ar PMS CPcdP, APA, ACEA, CPhiAPA, 

CPhiACEA 

 Yes 

19320 LaVoie et al., 1979 TA98, TA100 Rat Ar S9 BeP, Pery Yes Several other PAHs were evaluated, but 

concurrent with BaP. 

19360 LaVoie et al., 1985 TA98, TA100 Rat Ar S9 AC Yes 23650 McCann et al., 1975 TA1535, TA1537, 

TA98, TA100 

Rat Ar S9 DBaiP, BeP, DBacA, DBahA, CH, BaA Pyr, AC, PH, FE Yes 

15170 Norpoth et al., 1984 TA100 Rat and mouse S9; 

induction by Clophen A50 and 18 PAHs 

BaA No S9 composition was different for BaA an

BaP; result cannot be compared. 

20220 Pahlman and 

Pelkonen, 1987 

TA100 S9 from control, MC, 

or TCDD treated rats and mice 

BaA, CH, Tphen, DBacA, DBahA AN, AC, PH, 

FE, Pyr, BeP, Pery, PCE 

Yes 

20530 Penman et al., 1980 TM677 Rat Ar or PB S9 Pery, CPcdP, DBacA No No concurrent control values were repor20450 Phillipson and 

Ioannides, 1989 

TA100 S9 isolated from 

mouse, hamster, rat, 

pig, and human 

BaA, DBaiP, DBahA Yes 

20490 Poncelet et al., 1978 TA1530, TA1535, 

TA1537, TA1538, TA98, TA100 

S9 (origin unknown) CO, Tphen, FA, BghiP BbF No Qualitative data reported in published 

abstract. 

20560 Probst et al., 1981 TA1530, TA1535, 

TA1537, TA1538, TA98, TA100 

Rat Ar S9 BbA, DBacA AC, DBahA, 

PH, Pyr, DBaiP 

No Data reported as minimum mutagenic 

concentration (nmol/mL). 

20880 Rosenkranz and 

Poirier, 1979 

TA1530, TA1535 Uninduced rat S9 AC, BaA, BeP, 

CH, PH 

Yes 

21000 Sakai et al., 1985 TA97, TA98, TA100 Rat Ar S9 FE (equiv.), AC, PH, FA, CH, Pyr, BeP, 

Pery, BghiP, CO 

 Yes 

21040 Salamone et al., 

1979a 

TA1535, TA1537, TA1538, TA98, TA100 

Rat Ar S9 BaA, BeP (equiv), BghiP, DBaiP, BPH, 

CH, CO, DBacA, PCE 

AC, BaFE, BbFE, FA, Pery, Pyr 

No Increase in spontaneous mutation rate w

indicated, but dose data were not provid
78 DRAFT – DO NOT CITE OR QUO

Table 4-9. Study summaries: in vitro bacterial mutagenicity with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Salmonella strain(s) Activation system Positive result Negative result

Meets 

selection 

criteria? Comments 

13260 Salamone et al., 

1979b 

TA98, TA100 Rat Ar S9 DBaiP No Dose-response data were not completely 

reported; maximal response information (dose and number of revertants) was 

presented in text; BaP max response at 

different S9 than DBaiP. 

11860 Sangaiah et al., 

1983 

TA1535, TA1537, TA1538, TA98, TA100 

Rat Ar S9 BjAC Yes Dose-response data for BaP was present

for TA98 only. 

21360 Simmon, 1979a TA1535, TA1536, 

TA1537, TA1538, TA98, TA100 

Rat Ar S9 BaA, BeP AC, CH, PH Yes 

21640 Teranishi et al., 

1975 

TA1535, TA1536, TA1537, TA1538 

S9 from rats treated with PB and MC or DBahA 

DBaiP, DBaeP DBahA, BaA, 

BeP 

Yes 

16180 Utesch et al., 1987 TA100 Intact or 

homogenized hepatocytes from Ar treated rats 

BaA Yes 

16440 Wood et al., 1980 TA98, TA100 Rat Ar S9 and 

purified MFO 

enzymes system 

CPcdP Yes 

Ar = Arochlor 1254-treated; MC = 3-methylcholanthrene-treated; PB = phenobarbital-treated; PMS = postmitochondrial supernatant 
79 DRAFT – DO NOT CITE OR QUO

1 

Table 4-10. Study summaries: in vitro mammalian mutagenicity assays with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Cell type Metabolic activation Mutagenesis assay 

Positive 

result 

Negative 

result 

Meets selection 

criteria? Commen

16900 Allen-Hoffmann and 

Rheinwald, 1984 

Human epidermal keratinocyte 

None 6-Thioguanine resistance (HPRT) BaA Yes 

16920 Amacher and Paillet, 

1982 

Mouse lymphoma cells (L5178Y) 

Syrian golden hamster S9 mix or cocultivated hamster hepatocytes 

Trifluorothymidine resistance (thymidine kinase locus [TK]) 

BaA Yes 

16930 Amacher and Paillet, 

1983 

Mouse lymphoma cells (L5178Y) 

Cocultivated rat hepatocytes Trifluorothymidine resistance (TK) BaA Yes 

16940 Amacher and Turner, 

1980 

Mouse lymphoma cells (L5178Y) 

S9 from eight rodent species or strain; one rat strain induced by Ar 

Trifluorothymidine resistance (TK) AC, BaA Yes AC data no

useable; Banot simultaneou

16910 Amacher et al., 1980 Mouse lymphoma cells 

(L5178Y) 

Rat Ar and noninduced S9 Trifluorothymidine resistance (TK) BaA AC, Pyr Yes 

13440 Baird et al., 1984 V79 Chinese hamster cells Hamster embryo cells 6-Thioguanine resistance (HPRT) BeP Yes 17140 Barfknecht et al., 1982 TK6 human lymphoblast cells Rat Ar S9 Trifluorothymidine resistance (TK) FA, BaA, 

CH, Tphen, CPcdP 

PH, AC, ACEP 

Yes 

24670 Durant et al., 1999 H1A1v2 human 

lymphoblastoid cells 

Transfected with cyp1a1 cDNA Trifluorothymidine resistance (TK) BaPery, 

BbPery, DBaeF, DBafF, DBahP, DBaiP, DBelP, 

N23aP, N23eP 

DBjlF, 

N12bF 

Yes 

18260 Gehly et al., 1982 C3H/10T1/2 clone 8 mouse 

fibroblast cells 

None Ouabain resistance (HPRT) BeP Yes 

14250 Hass et al., 1982 V79 Chinese hamster cells Hamster embryo cells Ouabain and 6-thioguanine 

resistance (HPRT) 

DBaiP, DBahP 

 Yes 

18750 Huberman, 1975 V79 Chinese hamster cells Hamster cells 8-Azaguanine resistance (HPRT) BaA, Pyr Yes 18740 Huberman and Sachs, 

1976 

V79 Chinese hamster cells Hamster embryo cells Ouabain and 8-azaguanine 

resistance (HPRT) 

DBacA, DBahA (both weak)

Pyr, PH, CH, BaA 

Yes 

24120 Huberman and Sachs, 

1974 

V79 Chinese hamster cells Hamster embryo cells 8-Azaguanine resistance (HPRT) BaA Yes 

18990 Jotz and Mitchell, 

1981 

Mouse lymphoma cells (L5178Y) 

Rat Ar S9 Trifluorothymidine resistance (TK) Pyr Yes 

24720 Kligerman et al., 1986 Mouse lymphoma cells 

(L5178Y) 

Rat Ar S9 Trifluorothymidine resistance (TK) BlAC Yes 
80 DRAFT – DO NOT CITE OR QUO

Table 4-10. Study summaries: in vitro mammalian mutagenicity assays with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Cell type Metabolic activation Mutagenesis assay 

Positive 

result 

Negative 

result 

Meets selection 

criteria? Commen

19180 Krahn and 

Heidelberger, 1977 

V79 Chinese hamster cells Rat MC S9 6-Thioguanine resistance (HPRT) BaA, 

DBacA, DBahA 

 Yes DBacA and 

DBahA datnot useabletreatment different thaBaP. 

24680 Lafleur et al., 1993 MCL-3 human 

lymphoblastoid cells 

Transfected with cyp1a2 and cyp2a6 cDNA 

Trifluorothymidine resistance (TK) CPcdP, 

ACEA, CPhiACEA

APA, CPhiAPA, BghiF 

Yes 

24170 Langenbach et al., 

1983 

V79 Chinese hamster cells Cocultivation with primary rodent 

cells from liver, lung, kidney, and 

bladder 

Ouabain resistance (HPRT) AC Yes 

7550 Li and Lin, 1996 HS1 HeLa cells (human 

epithelial cells) 

None 6-Thioguanine resistance (HPRT) BaA Yes 

19870 Mishra et al., 1978 Fischer rat embryo cells 

infected with Rauscher leukemia virus 

Rat Ar S9 Ouabain resistance (HPRT) AC, PH, 

Pyr, BeP 

Yes 

20040 Myhr and Caspary, 

1988 

Mouse lymphoma cells (L5178Y) 

Rat Ar and noninduced S9 Trifluorothymidine resistance (TK) AC, BaA, 

BeP 

 No Results 

reported as ranges. 

11450 Nesnow et al., 1984 V79 Chinese hamster cells Rat Ar S9 6-Thioguanine resistance (HPRT) BlAC, 

BeAC, BjAC 

 Yes 

15630 Raveh and Huberman, 

1983 

V79 Chinese hamster cells Hamster embryo fibroblasts 6-Thioguanine resistance(HPRT); 

phorbol myristate acetate used to 

enhance recovery 

CPcdP BaA Yes 

15640 Raveh et al., 1982 V79 Chinese hamster cells Hamster embryo fibroblasts Ouabain and 6-thioguanine 

resistance (HPRT) 

CPcdP Yes Mutagenici

correlated wskin tumor initiation. 

21410 Slaga et al., 1978 V79 Chinese hamster cells Hamster embryo cells Ouabain resistance (HPRT) BaA (weak) Yes 21720 Tong et al., 1983 Rat liver epithelial cells 

(ARL-18) 

 6-Thioguanine resistance (HPRT) BaA, 

BeP, Pyr 

No Repeats dat

from 21730 Tong et al., 1981b 

21730 Tong et al., 1981b Rat liver epithelial cells

(ARL-18) 

None 6-Thioguanine resistance (HPRT) BeP, Pyr, 

BaA 

Yes 
81 DRAFT – DO NOT CITE OR QUO

Table 4-10. Study summaries: in vitro mammalian mutagenicity assays with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Cell type Metabolic activation Mutagenesis assay 

Positive 

result 

Negative 

result 

Meets selection 

criteria? Commen

16190 Vaca et al., 1992 UV-sensitive Chinese hamster 

ovary (CHO) cells 

Rat Ar S9 6-Thioguanine resistance (HPRT) FA Yes 

21900 Wangenheim and 

Bolcsfoldi, 1988 

Mouse lymphoma cells (L5178Y) 

Rat Ar S9 Trifluorothymidine resistance (TK) Pyr, FE Yes 

HPRT = hypoxanthine-guanine phosphoribosyl transferase mutagenicity assay (resistance to 6-thioguanine, 8-azaguanine, or ouabain); TK = thymidine kinase mutagenicity assay (resistance to 

trifluorothymidine) 
82 DRAFT – DO NOT CITE OR QUO

1 

Table 4-11. Study summaries: in vitro morphological/malignant cell transformation with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Cell type Metabolic activation system Positive result Negative result

Meets selection 

criteria? Comments 

13390 Atchison et al., 1985 BALB/3T3 mouse embryo fibroblasts None FA, Pyr Yes 17610 Casto, 1979 Syrian golden hamster embryo cells None DBahA Pyr Yes 17730 Chen and 

Heidelberger, 1969 

Adult C3H mouse ventral prostate cells Cocultivated irradiated C3H 

mouse embryonic fibroblasts 

DBahA DBacA, Pyr No Control data not provided. 

24750 Davis, 1999 C3H10T1/2 cells None DBalP, DBaeP, 

BcC, BgC, BcPH 

 No Control data not provided. 

17970 DiPaolo et al., 1969 Syrian golden hamster embryo cells Cocultivated irradiated 

Sprague-Dawley rat fetal cells 

DBahA, BaA, BeP, DBacA 

Pyr, PH Yes 

17990 DiPaolo et al., 1972 BALB/3T3 None AC, Pyr Yes 23630 DiPaolo et al., 1973 Syrian golden hamster embryo cells In vivo (transplacental) 

exposure 

 AC, PH, Pyr No No quantitative information. 

18020 Dunkel et al., 1981 Balb/3T3, Syrian golden hamster embryo, 

and Rauscher murine leukemia virusinfected F344 rat embryo cells 

None BaA BeP, PH, AC Yes Qualitative data only for R-MuL

RE cells. BaA positive in SHEMequivocal in Balb/3T3. 

18080 Emura et al., 1980 Syrian golden hamster fetal lung cells None BbF, BaA, IP BkF, BeP Yes 23640 Evans and DiPaolo, 

1975 

Strain 2 guinea pig fetal cells None AC, Pyr, PH No No quantitative information. 

18260 Gehly et al., 1982 C3H10T1/2CL8 mouse embryo fibroblasts None BeP Yes 14130 Greb et al., 1980 BHK 21/CL 13 Rat Ar S9 CH, BaA, BbF, 

DBahA, BeP 

PH, AC Yes 

23890 Kakunaga, 1973 BALB/3T3 subclone A31-714 None PH, Pyr No Not clear if BaP admin 

simultaneously. 

14640 Krolewski et al., 1986 C3H10T1/2CL8 mouse embryo fibroblasts None CPcdP Yes 14700 Laaksonen et al., 1983 Newborn NMRI nu/nu nude mouse skin 

fibroblasts 

None BaA AC Yes 

14850 Lubet et al., 1983 C3H10T1/2CL8 mouse embryo fibroblasts None BeP AC, DBahA, PH Yes 19870 Mishra et al., 1978 Rauscher leukemia virus-infected Fischer 

rat embryo 

None AC, PH, Pyr, 

BeP 

No No quantitative information. 

24710 Mohapatra et al., 1987 C3H10T1/2CL8 mouse embryo fibroblasts None BeAC, BjAC, 

BlAC 

BkAC Yes 

24700 Nesnow et al., 1990 Human neonatal foreskin fibroblasts None BlAC Yes 7980 Nesnow et al., 1997 C3H10T1/2CL8 mouse embryo fibroblasts None DBalP Yes 7990 Nesnow et al., 1994 C3H10T1/2CL8 mouse embryo fibroblasts None DBahA Yes 8000 Nesnow et al., 1993a C3H10T1/2CL8 mouse embryo fibroblasts None DBkmnoAPH DBjmnoAPH, 

N123mnoAPH 

Yes 

20120 Nesnow et al., 1991 C3H10T1/2CL8 mouse embryo fibroblasts None ACEA Yes 
83 DRAFT – DO NOT CITE OR QUO

Table 4-11. Study summaries: in vitro morphological/malignant cell transformation with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Cell type Metabolic activation system Positive result Negative result

Meets selection 

criteria? Comments 

23720 Pienta et al., 1977 Syrian golden hamster embryo Cocultivated X-irradiated 

cells of same type 

BaA, DBahA CH, BeP, Pyr, 

AC, DBacA, PH

Yes 

8490 Sheu et al., 1994 BALB/3T3 A31-1-1 None Pyr, BaA, CH Yes 
84 DRAFT – DO NOT CITE OR QUO

1 

Table 4-12. Study summaries: in vitro DNA adducts with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Cell type or DNA source 

Incubation 

time Activation system Method of analysis

PAHs 

evaluated

a

Meets selection 

criteria? Comments 

16890 Allen and Coombs, 

1980 

Mouse embryo cells from TO mice 24 hr None [

3H] prelabeling BaA Yes 

6300 Binkova et al., 2000 Human diploid lung fibroblast 

cells 

Various up to 24 hr 

None [32P] postlabeling DBalP Yes 

9510 Bryla and Weyand, 

1992 

Calf thymus DNA 1 hr None [32P] postlabeling BaA, DBacA, 

PH 

Yes PH did not form measurable 

DNA adducts. Adduct formation enhanced when reacted under white light. 

6570 Cherng et al., 2001 Human hepatoma HepG2 cells 24 hr None [32P] postlabeling BghiP Yes BghiP did not form measura

DNA adducts. 

13780 Cooper et al., 1982 Fibroblasts and epithelial cells 

from Wistar rat mammary tissue 

24 hr None [

3H] prelabeling BaA Yes BaA formed little or no 

measurable DNA adducts. 

22800 Grover and Sims, 1968 Salmon testes DNA Not specified Rat liver microsomes [

3H] prelabeling DBahA, DBacA, 

BaA, Pyr, PH 

Yes 

10660 Johnsen et al., 1998 Human lymphocytes and human 

promyelocytic HL-60 cells 

24 hr None [32P] postlabeling BjAC, BlAC Yes 

10670 Johnsen et al., 1997 Rat lung Clara cells, Type 2 cells, 

and macrophages 

2 hr PCB pretreatment of 

whole animals 

[32P] postlabeling BjAC, BlAC Yes 

13200 Li et al., 2002 MCF-7 cells or rat lung DNA 7–24 hr Human mammary 

microsomes with rat lung DNA 

[32P] postlabeling DBalP, BcPH, 

DBahA 

No No quantitative results. 

7870 Melendez-Colon et al., 

2000 

Human mammary carcinoma MCF-7 cells and leukemia HL-60 cells 

4 or 24 hr None [32P] postlabeling DBalP Yes No adducts formed in HL-60 

cells that lack significant P4activity. 

7990 Nesnow et al., 1994 C3H10T1/2CL8 fibroblasts 24 hr None 32P] postlabeling DBahA No No quantitative results. 20120 Nesnow et al., 1991 C3H10T1/2 cells 24 hr None 32P] postlabeling ACEA No Measures repair of adducts o

not synthesis. 

21200 Segerback and 

Vodicka, 1993 

Calf thymus DNA 3 hr Rat Ar S9 32P] postlabeling, 

3Hbinding CH, BaA, BbF, DBahA, FA, 

BghiP, Pyr 

Yes 

24810 Baird et al., 2002 MCF-7 cells 24 hr Morpholinos 

inhibition (antisense oligomer that blocks 

protein synthesis of 

CYPIA1) 

[32P] postlabeling DBalP No Confounded by CYP1A1 

inhibition by morpholinos. 

aExcept where noted, positive findings were reported for all PAHs evaluated. 
85 DRAFT – DO NOT CITE OR QUO

1 

Table 4-13. Study summaries: in vitro DNA damage, repair, or synthesis with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Cell type Metabolic activation Endpoint Assay Positive result 

Negative 

result 

Meets selection 

criteria? Comment

16840 Agrelo and Amos, 

1981 

Human fibroblasts Rat Ar S9 Unscheduled DNA 

synthesis 

[

3H] Thymidine 

uptake 

Pyr Yes 

17610 Casto, 1979 Syrian golden hamster 

embryo 

Intrinsic Unscheduled DNA 

synthesis 

[

3H] Thymidine 

uptake 

 DBahA, Pyr, 

PH 

Yes 

24030 De Flora et al., 1984 Escherichia coli WP2, 

WP67, and CM871 

Rat Ar S9 DNA damage Differential 

killing repairdeficient strains 

AC, BaA Pery, BeP No Semiquantita

data. 

18030 Dunkel et al., 1984 E. coli WP-2 uvrA Rat, mouse, hamster 

Ar S9 

DNA damage Differential 

killing repairdeficient strains 

BaA, BeP, PH, Pyr AC No Dose-respons

data not 

provided. 

23790 Ichinotsubo et al., 

1977 

E. coli Rec BC S9 (origin unknown) DNA damage DBaiP, DBahA Yes 

10670 Johnsen et al., 1997 Rat lung Clara cells, Type 2 

cells, and macrophages 

PCB pretreatment of whole animals 

DNA damage Alkaline elution BjAC, BlAC No No untreated 

control. 

10660 Johnsen et al., 1998 Human lymphocytes and 

human promyelocytic HL60 cells 

Rat or human liver microsomes 

DNA damage Alkaline elution BjAC, BlAC Yes 

19270 Lake et al., 1978 Human foreskin epithelial 

cells 

None Unscheduled DNA 

synthesis 

[

3H] Thymidine 

uptake 

DBahA AC, BeP, PH, 

Pyr 

No Doses reporte

as ranges. 

19680 Mamber et al., 1983 E. coli WP2 and WP100 Rat Ar S9 DNA damage Growth 

inhibition of repair deficient strains 

 AC, FE, Pyr Yes 

19690 Mane et al., 1990 Human and rat mammary 

epithelial cells 

None Inhibition of DNA 

synthesis 

[

3H] Thymidine 

uptake 

BaA (in human MEC only) 

BeP No Positive respo

for BaA not observed consistently. 

19730 Martin and 

McDermid, 1981 

HeLa S3 cells PB-induced rat liver 

postmitochondrial 

supernatant 

Unscheduled DNA synthesis 

[

3H] Thymidine 

uptake 

Pyr (authors: “dubious” result) 

AC No No quantitati

information. 

19740 Martin et al., 1978 HeLa S3 cells 3-MC induced rat 

liver 

postmitochondrial 

supernatant 

Unscheduled DNA synthesis 

[

3H] Thymidine 

uptake 

BeP, BaA, DBacA, DBahA 

Pyr, AC Yes 

23800 McCarroll et al., 

1981 

E. coli WP2, WP2 uvrA, 

WP67, CM611, WP100, W3110polA+, and 

p3478polaRat Ar S9 DNA damage Differential killing repair- deficient strains AC, PH Yes 
86 DRAFT – DO NOT CITE OR QUO

Table 4-13. Study summaries: in vitro DNA damage, repair, or synthesis with benzo[a]pyrene and at least one other PAH 

Record 

number Reference Cell type Metabolic activation Endpoint Assay Positive result 

Negative 

result 

Meets selection 

criteria? Comment

19830 MerschSundermann et al., 

1992 

E. coli PQ37 Rat Ar S9 Induction of SOS 

system 

SOS chromotest AA, BaA, BbF, BghiF, 

BjF, BbFE, BghiP, BeP, CH, DBacA, DBahA, DBalP, DBahP, DBaiP, FA, IP, PH, Tphen 

AC, BaFE, CO, FE, Pery, Pyr 

Yes 

19850 Milo et al., 1978 Human skin fibroblast NF 

and Detroit 550 cells 

None DNA damage Alkaline elution AC, Pyr, PH, 

BeP 

Yes 

20050 Nagabhushan et al., 

1990 

Hamster buccal pouch epithelial cells and tissue fragments 

Not specified Inhibition of DNA 

synthesis 

[

3H] Thymidine 

uptake 

 BaA No Abstract only

BaA inhibited synthesis 4%

20560 Probst et al., 1981 Rat hepatocyte primary 

culture 

None Unscheduled DNA 

synthesis 

[

3H] Thymidine 

uptake 

BbA, DBacA AC, DBahA, 

PH, Pyr, DBaiP, FE, BeP 

No Artifact of 

counting metresulted in control resporeported as negative valu

20810 Robinson and 

Mitchell, 1981 

Human fibroblasts WI-38 cells 

Rat Ar S9 Unscheduled DNA 

synthesis 

[

3H] Thymidine 

uptake 

Pyr (with activation) Yes 

23900 Rosenkranz and 

Leifer, 1980 

E coli pol A1- Rat liver S9 DNA damage Differential 

killing repairdeficient strains 

 AC, BaA, BeP, 

CH, PH 

Yes 

20880 Rosenkranz and 

Poirier, 1979 

E. coli pol A1- Uninduced rat S9 DNA damage Differential 

killing repairdeficient strains 

 AC, BaA, BeP, 

CH, PH 

Yes 

20940 Rossman et al., 

1991 

E coli WP2s(λ) Rat liver S9 DNA damage Λ prophage 

induction 

AC, DBacA, DBahA, PH 

BeP, FA, Pyr Yes 

21380 Simmon, 1979b Saccharomyes cerevisiae

D3 

Rat Ar S9 induced 

recombination 

Colony 

pigmentation on 

adenine medium

 AC, BaA, BeP, 

CH, PH 

Yes 

21720 Tong et al., 1983 Rat hepatocyte primary 

culture 

None Unscheduled DNA 

synthesis 

[

3H] Thymidine 

uptake 

BaA BeP, AC, CH, 

Pyr 

No Repeats data 

from 21730 Tet al., 1981b. 

21730 Tong et al., 1981b Rat hepatocyte primary 

culture 

None Unscheduled DNA 

synthesis 

[

3H] Thymidine 

uptake 

BaA BeP, AC, CH, 

Pyr 

Yes 

21790 Tweats, 1981 E. coli WP2, WP67(uvrA 

polA), CM871 (uvrA lexA 

recA) 

Rat Ar S9 DNA damage Differential 

killing repairdeficient strains 

 Pyr, AC No No quantitati

information. 

16190 Vaca et al., 1992 CHO cells Rat Ar S9 DNA damage Alkaline elution FA No No untreated 

vehicle contr

22260 Williams et al., 

1982 

Rat hepatocyte primary culture 

None Unscheduled DNA 

synthesis 

[

3H] Thymidine 

uptake 

 Pyr, BeP No No quantitati

information. 
87 DRAFT – DO NOT CITE OR QUO

1 

Table 4-14. Study summaries: in vitro clastogenicity or sister chromatid exchange with benzo[a]pyrene and at 

least one other PAH 

Record 

number Reference Cell type Metabolic activation 

Clastogenic 

endpoint(s) 

Positive 

results 

Negative 

results 

Meets selection 

criteria? Comments 

16740 Abe and Sasaki, 1977 Pseudodiploid Chinese 

Hamster D-6 

None Aberrations and SCEs AC, Pyr Yes 

17890 Dean, 1981 Near-diploid epithelialtype rat liver RL1 None Various aberrations AC, Pyr No Semiquantitative results. 17930 DeSalvia et al., 1988 Male Chinese hamster 

liver epithelial cells (CHEL) 

None SCEs Pyr, FA Yes 

18120 Evans and Mitchell, 

1981 

CHO Rat Ar S9 SCEs Pyr (with 

activation) 

 No No untreated or vehicle control. 

23640 Evans, and DiPaolo, 

1975 

Diploid strain 2 guinea 

pig fetal cells 

None Aneuploidy AC No No quantitative data. Pyr, PH also 

evaluated using different protocol withoBaP reference. 

18260 Gehly et al., 1982 CH3/10T1/2 Clone 8 

mouse fibroblasts 

None SCEs BeP Yes 

14620 Kochhar, 1982 Chinese hamster V79 None Aberrations including 

gaps, rings, breaks, fragments, exchanges 

BaA Yes Dose-dependent increase in % cells wit

aberrations. 

14640 Krolewski et al., 1986 CH3/10T1/2 Clone 8 

mouse embryo cells 

None SCEs CPcdP Yes CPcdP appears to increase SCEs in dos

dependent fashion (two doses). 

19690 Mane et al., 1990 Chinese hamster V79 

cells 

With and without rat mammary epithelial cell coculture SCEs BaA BeP Yes 

19770 Matsuoka et al., 1979 Male Chinese hamster 

lung (CHL) 

Rat Ar S9 Aberrations and SCEs PH No Not clear if BaP administered simultane

ously. No untreated control. 

20020 Murison, 1988 P3 clonal isolate from 

human epithelial teratocarcinoma 

BJ-015 human breast epithelial cell coculture 

SCEs CPcdP BeP No Not clear if BaP administered 

simultaneously; no concurrent control. 

20340 Perry and Thomson, 

1981 

CHO cells Rat Ar S9 SCEs Pyr AC No No untreated control. 
88 DRAFT – DO NOT CITE OR QUO

Table 4-14. Study summaries: in vitro clastogenicity or sister chromatid exchange with benzo[a]pyrene and at 

least one other PAH 

Record 

number Reference Cell type Metabolic activation 

Clastogenic 

endpoint(s) 

Positive 

results 

Negative 

results 

Meets selection 

criteria? Comments 

20500 Popescu et al., 1977 Chinese hamster 

V79-4 cells 

With or without irradiated Syrian golden hamster secondary embryo feeder cells

Aberrations and SCEs Pery, Pyr PH No BaP increased SCEs but Pyr and Pery i

creased aberrations. Pery increased aberations w/o activation. 60% of Pyr treacells (activated) polyploid. Increased aberrations in polyploid cells. 

21710 Tong et al., 1981a Adult rat liver 

epithelial (ARL 18) cells 

None SCEs BaA BeP, Pyr, 

AC 

Yes 

21720 Tong et al., 1983 Adult rat liver 

epithelial (ARL 18) cells 

None SCEs BaA BeP, Pyr, 

AC 

No Repeats data from 21710 Tong et al., 

1981a 

8780 Vienneau et al., 1995 UDP-Glucuronosyltransferases-deficient 

rat (RHA-J/J) skin 

fibroblasts 

None Micronuclei BeP Yes 

8850 Warshawsky et al., 

1995 

Human lymphocytes None Micronuclei and SCEs BaA Yes 

21980 Weinstein et al., 1977 Human diploid 

fibroblasts (WI-38) 

With or without rat Ar s9 Chromosomal 

damage, mitotic index, abnormal metaphases 

 Pyr Yes 
89 DRAFT – DO NOT CITE OR QUOTE 

1 If the above criteria were met, studies were selected for use in the analysis regardless of 

2 whether positive or negative results were reported. Studies with positive findings were used for 

3 calculation of RPFs. Studies with negative findings were used in a weight of evidence 

4 evaluation of potential carcinogenicity (discussed later in Section 6.1). To be considered 

5 adequate for use in the analysis, nonpositive bioassays were selected only if two additional 

6 conditions were met: (1) at least 20 animals were used per dose group, and (2) animals were 

7 observed for at least 6 months. More strict criteria were applied to nonpositive studies due to the 

8 difficulty in demonstrating the absence of an effect. For example, if a positive tumor response 

9 (i.e., statistically significant increase in incidence) was observed after 3 months of treatment with 

10 a given PAH, the positive finding is clear; however, if no response (or a nonsignificant response) 

11 was observed after 3 months, the absence of response might reflect a lack of carcinogenic action, 

12 but might also have resulted from inadequate follow-up time. The use of these additional criteria 

13 for nonpositive studies served to ensure that PAHs would not be treated as noncarcinogenic 

14 based on inadequate nonpositive bioassays. 

15 Study design details, findings, limitations, and a determination of whether the study met 

16 selection criteria are presented in Tables 4-1 through 4-14 for each study reviewed in each 

17 category. Positive and negative findings as reported in the table are based on the author’s 

18 determination except where noted. When statistical analysis of tumor bioassay data was not 

19 included in the pertinent publication, statistical analysis was conducted to determine whether the 

20 response differed from control. In the sections that follow, overviews of the data available in 

21 each category are presented. The overviews address the nature of the studies available, concise 

22 information on general study methods, general findings for the tested compounds, and key 

23 strengths and limitations of the available data for relative potency development. 

24 

25 4.3.1. In Vivo Cancer Bioassays in Animals 

26 The PAH database contained a large number of cancer bioassay studies in which one or 

27 more PAHs was evaluated along with benzo[a]pyrene. The vast majority of the tumor bioassay 

28 studies were mouse skin painting studies (n = 43). In addition, there were 11 intraperitoneal 

29 studies, 9 subcutaneous exposure studies, 2 oral studies, and 9 studies using miscellaneous 

30 exposure routes. 

31 

32 4.3.1.1. Dermal Exposure 

33 A summary of the 43 dermal bioassays is provided in Table 4-1. These studies were all 

34 conducted in mice. Fifteen studies tested the complete carcinogenicity of PAHs, while 

35 23 studies tested PAHs as initiators in initiation-promotion protocols. In some cases, both 

36 complete and initiation-promotion studies were reported in the same reference. For these 

37 references, two entries are included in the table. 
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1 Complete carcinogenicity studies were conducted in mice using either dropper or 

2 paintbrush application. Swiss mice were typically preferred for these studies. PAHs, usually in 

3 acetone, were applied to the shaved interscapular skin 2 or 3 times/week. The duration of 

4 exposure varied from 10 weeks up to about 70 weeks; most studies continued exposure for at 

5 least 30 weeks. Skin tumor counts were recorded on a weekly basis, and animals were sacrificed 

6 when tumors reached a minimum size (e.g., 2 cm) or when the animals were moribund. These 

7 studies generally focused exclusively on skin papillomas and carcinomas. Skin tumor data were 

8 reported as incidence (i.e., number of animals with tumors) and/or tumor count (mean number of 

9 tumors per animal) (indicated in Table 4-1). 

10 Several PAHs consistently (in two or more studies) proved to be complete carcinogens in 

11 mouse skin painting assays, including benzo[b]fluoranthene, benzo[j]fluoranthene, 

12 cyclopenta[c,d]pyrene, dibenzo[a,e]pyrene, dibenzo[a,h]pyrene, dibenzo[a,i]pyrene, and 

13 dibenzo[a,l]pyrene. Chrysene gave positive results in two complete carcinogenicity studies 

14 (LaVoie et al., 1979; Wynder and Hoffmann, 1959) and equivocal results in a third (Hecht et al., 

15 1974). Anthanthrene, dibenzo[a,e]fluoranthene, and dibenz[a,h]anthracene each gave positive 

16 tumorigenicity results in a single assay (Cavalieri et al., 1977; Hoffmann and Wynder, 1966; and 

17 Wynder and Hoffmann, 1959; respectively). Negative or equivocal results were reported for 

18 benzo[k]fluoranthene, benzo[g,h,i]fluoranthene, dibenzo[e,l]pyrene, indeno[1,2,3-c,d]pyrene, 

19 benzo[g,h,i]perylene, naphtho[2,3-e]pyrene, anthracene, pyrene, fluoranthene, 2,3-acepyrene, 

20 benz[a]anthracene, coronene, and benzo[e]pyrene (see Table 4-1). 

21 According to LaCassagne et al. (1968), in studies conducted prior to 1966, the compound 

22 reported as dibenzo[a,l]pyrene was actually dibenzo[a,e]fluoranthene. In the text and tables of 

23 this report, data from Hoffmann and Wynder (1966) are reported as dibenzo[a,e]fluoranthene in 

24 Table 4-1. 

25 The initiation studies in Table 4-1 were performed under a generally consistent protocol, 

26 as follows. During the early part of the second telogen phase of the hair cycle (at about 7–8 

27 weeks of age), PAHs in acetone were applied to the shaved interscapular skin of mice. In 

28 general, female Swiss, CD-1, or SENCAR mice were used. Some studies used dropper 

29 administration, but the majority employed a painting method using a camel’s hair brush. About 

30 half of the initiation studies used a single initiation dose, while the other half administered the 

31 initiating compound in 10 subdoses given every other day. One to 2 weeks after the final 

32 initiating dose, promotion was begun with twice or thrice weekly applications of a promoting 

33 agent, usually TPA (12-0-tetra-decanoylphorbol-13-acetate) or croton oil. The dose of the 

34 promoting agent varied by study. Promotion usually continued for about 20 weeks (with a range 

35 across studies from 11 to 26 weeks). The incidence of skin papillomas was recorded on a weekly 

36 basis until the promotion period was ended. Papillomas were removed at random for histological 

37 verification. Some studies reported the number of tumors per animal; some reported only the 

38 incidence. 
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1 The initiation studies in Table 4-1 consistently showed positive tumorigenicity across two 

2 or more studies for the following compounds: benzo[j]fluoranthene, benzo[b]fluoranthene, 

3 chrysene, dibenzo[a,e]pyrene, dibenzo[a,h]pyrene, dibenzo[a,i]pyrene, dibenzo[a,l]pyrene, and 

4 cyclopenta[d,e,f]chrysene. In at least one study, benzo[k]fluoranthene, benz[l]aceanthrylene, 

5 benz[e]aceanthrylene, naphtho[2,3-e]pyrene, dibenz[a,h]anthracene, dibenz[a,c]anthracene, and 

6 benz[b,c]aceanthrylene showed positive initiating activity. Negative results were reported for 

7 pyrene, perylene, benzo[g,h,i]fluoranthene, fluoranthene, anthanthrene, dibenzo[e,l]pyrene, 

8 benzo[g,h,i]perylene, indeno[1,2,3-c,d]pyrene, benzo[e]pyrene, anthracene, 2,3-acepyrene, and 

9 phenanthrene. Cyclopenta[c,d]pyrene gave negative results in one study—Wood et al. (1980)—

10 and positive results in two— Cavalieri et al. (1981b) and Raveh et al. (1982) (see Table 4-1). 

11 The vast majority of the initiation and complete carcinogenicity studies were conducted 

12 in female mice, so data on gender differences in skin tumor susceptibility are not available. 

13 A few studies using dermal application (Warshawsky et al., 1993; Slaga et al., 1979; Van 

14 Duuren and Goldschmidt, 1976; Horton and Christian, 1974; Van Duuren et al., 1973) were 

15 designed to evaluate the cocarcinogenicity of two or more PAHs, or of a single PAH with 

16 dodecane as a vehicle. These were primarily complete carcinogenicity studies, wherein PAHs 

17 were administered together over a chronic time period, although Slaga et al. (1979) used an 

18 initiation-promotion design. Study design was similar to other complete carcinogenicity 

19 experiments. In these studies, the carcinogenicity of single PAHs was evaluated for comparison 

20 with the results obtained when the PAHs were administered with a cocarcinogen. Data on single 

21 PAHs (without a cocarcinogen) were generally limited to single dose levels. In the 

22 cocarcinogenesis studies, only dibenz[a,c]anthracene, benzo[e]pyrene, and pyrene gave positive 

23 results when administered without a cocarcinogen; results for pyrene were judged to be 

24 equivocal in the absence of statistical confirmation. The PAHs chosen for cocarcinogenesis 

25 studies were often those traditionally understood to be nontumorigenic or weakly tumorigenic 

26 when administered alone (e.g., perylene, pyrene, benzo[e]pyrene, benzo[g,h,i]perylene, 

27 phenanthrene, fluoranthene). 

28 Several issues relating to the potential use of the dermal bioassay data for relative 

29 potency development were identified during study review. Several studies did not include a 

30 concurrent untreated or vehicle-treated control group (Masuda and Kagawa, 1972; Bingham and 

31 Falk, 1969; Wynder and Hoffmann, 1959a, b). In a number of reports, it appears that bioassays 

32 were done in batches and reported in a single publication. In these cases, it appears that 

33 benzo[a]pyrene treatment may not have been undertaken concurrently with all of the compounds 

34 in the report. For some of these studies (Horton and Christian, 1974; Bingham and Falk, 1969), 

35 there are differences in the choice of vehicle or promoter, or other issues that argue against using 

36 the benzo[a]pyrene data for direct comparison. In several others studies, however (Rice et al., 

37 1988; Slaga et al., 1980; Van Duuren and Goldschmidt, 1976; Wynder and Hoffmann, 1959), the 

38 protocols (including vehicle and promoting agent) appear to have been the same. 
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1 Among the dermal tumor bioassay studies in Table 4-1, 24 studies met the selection 

2 criteria for use in this analysis.

3 

4 4.3.1.2. Intraperitoneal Exposure 

5 Eleven cancer bioassays in the literature used intraperitoneal injection. Six of these 

6 studies were carried out in newborn mice, while the other five used adult A/J mice. All of the 

7 studies focused on lung and liver tumorigenicity after PAH exposure. Study summaries for all of 

8 these references are reported in Table 4-2. Tumor data were reported as incidence (i.e., number 

9 of animals with tumors) and/or tumor count (mean number of tumors per animal) (indicated in 

10 Table 4-2). 

11 Newborn mouse studies. Six cancer bioassays in newborn mice were identified (LaVoie 

12 et al., 1994, 1987; Busby et al., 1989, 1984; Weyand and LaVoie, 1988; Wislocki et al., 1986). 

13 In general, PAHs were administered intraperitoneally to newborn mice (usually of the Swiss or 

CD-1 strains). The dosing schedule called for 1/7th, 2/7ths, and 4/7ths 14 of the total dose to be 

administered on the 1st, 8th, and 15th 15 days of life. Typically, the mice were sacrificed at either 6 

16 months or 1 year, and lung and/or liver tumors were identified and classified. 

17 The studies in newborn mice showed a distinct gender difference in liver tumorigenicity. 

18 Male mice appear to be substantially more susceptible to liver tumor induction than females. In 

19 contrast, both male and female mice developed lung tumors after exposure. Three studies 

20 (LaVoie et al., 1994; Busby et al., 1989, 1984) reported that fluoranthene induced lung tumors in 

21 both male and female mice, while one study reported that fluoranthene induced liver tumors in 

22 male mice only (LaVoie et al., 1994). LaVoie et al. (1987) reported that benzo[b]fluoranthene 

23 and benzo[j]fluoranthene induced lung adenomas in both male and female mice, but induced 

24 liver tumors only in males. Wislocki et al. (1986) reported that treatment with benz[a]anthracene 

25 resulted in a significant increase in liver tumors in male mice. In this study, benz[a]anthracene 

26 treatment resulted in an increased incidence of lung tumors in both males and females, although 

27 the tumor incidence was significantly increased only for females. The same authors (Wislocki et 

28 al., 1986) reported a significant increase in liver tumors in male mice treated with chrysene, but 

29 no increase in lung tumorigenicity. The lack of lung tumorigenicity in mice treated with 

30 chrysene was also reported by Busby et al. (1989). 

31 Negative tumorigenicity results in newborn mouse assays were reported for pyrene, 

32 chrysene, benzo[k]fluoranthene, and indeno[1,2,3-c,d]pyrene (Busby et al., 1989; LaVoie et al., 

33 1987). 

34 Most of the data from the newborn mouse assays met the criteria for relative potency 

35 development, although Weyand and LaVoie (1988) is an abstract and does not provide dose36 response information. LaVoie et al. (1994) noted that liver tumorigenicity in newborn mice 

37 exposed to weak tumorigenic agents may not be fully realized for 12 months; thus, the failure to 
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1 observe liver tumors in studies of shorter duration (Busby et al., 1989, 1984) may result from the 

2 longer latency and should be taken into consideration in using these data. 

3 Lung adenoma A/J mouse studies. Five studies (Nesnow et al., 1998a, b, 1996, 1995; 

4 Ross et al., 1995; Mass et al., 1993) were carried out in 6- to 8-week-old A/J mice by the same 

5 laboratory using a standard protocol (Table 4-2). Mice were given a single intraperitoneal 

6 injection of PAH in tricaprylin and followed for 8 months. Upon sacrifice, the lungs were 

7 removed and adenomas were counted. Tumor multiplicity was reported, while tumor incidence 

8 was not. Several of these studies include estimates of relative potency based on statistical 

9 analysis of the tumor multiplicity data. 

10 These studies report positive tumor findings (reported as an increase in the number of 

11 tumors per animal) for all of the PAHs tested (benz[j]aceanthrylene, benzo[b]fluoranthene, 

12 dibenz[a,h]anthracene, cyclopenta[c,d]pyrene, and dibenzo[a,l]pyrene). 

13 Among the intraperitoneal tumor bioassay studies in Table 4-2, eight studies met the 

14 selection criteria for use in this analysis.

15 

16 4.3.1.3. Subcutaneous Injection Exposure 

17 Nine studies employing a subcutaneous exposure design were identified. All of the 

18 subcutaneous exposure studies are more than 25 years old; the most recent is Pfeiffer (1977). 

19 Study descriptions are presented in Table 4-3. 

20 Two studies utilized newborn mice (Roe and Waters, 1967; Grant and Roe, 1963). In 

21 these studies, phenanthrene was administered subcutaneously to newborn albino mice on the first 

22 day of life. Ten mice of each group were sacrificed after 52 weeks, and the remaining animals 

23 were sacrificed at 62 weeks. Grant and Roe (1963) reported lung tumorigenicity, while Roe and 

24 Waters (1967) reported liver tumors in the same group of mice. Roe and Waters (1967) reported 

25 an elevated incidence of liver tumors in male mice exposed subcutaneously to phenanthrene; 

26 however, it is not clear whether the difference was significant. Roe and Waters (1967) is a brief 

27 communication with limited details of the study design and results. 

28 In most of the remaining studies, single subcutaneous doses of one or more PAHs and 

29 benzo[a]pyrene were administered to mice, followed 1–2.5 years later by an evaluation of 

30 injection site and other tumors. Tumors at the injection site were most commonly reported; 

31 however, in some studies, investigators also examined other organs for tumors (Homburger et 

32 al., 1972; Roe and Waters, 1967; Grant and Roe, 1963; Rask-Nielsen, 1950; Pfeiffer and Allen, 

33 1948). 

34 Most of the subcutaneous bioassays suffer from critical shortcomings in design or 

35 reporting. One study used “aged” mice for controls, allowing these animals to live 16 weeks 

36 longer than the treated group (Homburger et al., 1972). Three studies gave apparently positive 

37 results for dibenz[a,h]anthracene (i.e., substantial tumor induction; Pfeiffer, 1977; Steiner, 1955; 

38 Bryan and Shimkin, 1943). However, neither Bryan and Shimkin (1943) nor Steiner (1955) 
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1 included untreated control groups. Pfeiffer (1977) included an untreated control group in which 

2 there was 90% mortality prior to sacrifice of the treated animals; data on tumor incidence in 

3 controls were not reported. Several other studies (Pfeiffer and Allen, 1948; Barry et al., 1935) 

4 also did not include a concurrent untreated or vehicle-treated control group. These studies were 

5 not used for dose-response assessment due to the lack of appropriate controls. 

6 Fundamental flaws were observed in two older studies. Pfeiffer and Allen (1948) 

7 examined the effects of PAHs in Rhesus monkeys. Individual animals were exposed 

8 sequentially to several PAHs via multiple exposure routes; thus, the effect of any individual PAH 

9 or benzo[a]pyrene cannot be discerned. Barry et al. (1935) treated mice with PAHs from varying 

10 sources and of varying purity. Given the age of the study and the attendant issues with 

11 nomenclature, purity, and analysis of the treatment compounds, data from this study are excluded 

12 from use in relative potency development. 

13 Among the subcutaneous tumor bioassay studies in Table 4-3, only a single study met 

14 selection criteria for use in this analysis.

15 

16 4.3.1.4. Oral Exposure 

17 The literature search identified two oral bioassays that included benzo[a]pyrene and at 

18 least one other PAH. Critical aspects of the study design for these studies are reported in 

19 Table 4-4. 

20 Biancifiori and Caschera (1962) compared the induction of mammary tumors in virgin 

21 and pseudopregnant mice (female mice mated with vasectomized males) after gavage exposure 

22 to dibenz[a,h]anthracene or benzo[a]pyrene. Tumor incidence was increased in pseudopregnant 

23 mice given 1 mg/week of either compound for 15 weeks, but not in virgin mice given the same 

24 dose. The relevance of the positive findings in pseudopregnant mice is uncertain given that an 

25 increased incidence of tumors was not observed in virgin mice treated at the same dose. One 

26 possible explanation for the disparate findings is that circulating hormones in pseudopregnant 

27 mice differed from those in virgin mice and interacted with the PAH to enhance tumor 

28 formation. Huggins and Yang (1962) also evaluated mammary tumor incidence after a single 

29 oral PAH exposure. Sprague-Dawley rats were given gavage doses of benzo[a]pyrene, 

30 benz[a]anthracene, or phenanthrene. This study did not include an untreated or vehicle-treated 

31 control group. No tumors were observed in the rats treated with either benz[a]anthracene or 

32 phenanthrene, while mammary tumors were observed in eight of the nine benzo[a]pyrene-treated 

33 animals. 

34 Among the oral tumor bioassay studies in Table 4-4, none met the selection criteria for 

35 use in this analysis.

36 
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1 4.3.1.5. Other Routes 

2 Nine bioassays were available that did not fit into other exposure route categories (i.e., 

3 dermal, intraperitoneal, subcutaneous, or oral) (see Table 4-5). Among these were studies using 

4 intramammillary, intramuscular, and intravenous injection as well as lung implantation, tracheal 

5 implantation, and transplacental exposure after subcutaneous injection. Seven studies were in 

6 rats, with one each in mice and hamsters. 

7 Deutsch-Wenzel et al. (1983) and Wenzel-Hartung et al. (1990) implanted 

8 PAH-containing pellets (consisting of beeswax and trioctanoin) into the lungs of inbred female 

9 Osborne-Mendel rats. Lung tumor incidence was reported for a total of 10 PAHs and 

10 benzo[a]pyrene. The authors reported relative potency estimates based on the lung tumor data. 

11 Lung tumors were induced by benzo[b]fluoranthene, benzo[j]fluoranthene, 

12 benzo[k]fluoranthene, benzo[g,h,i]perylene, indeno[1,2,3-c,d]pyrene, anthanthrene, chrysene, 

13 and dibenz[a,h]anthracene. Negative findings were reported for benzo[e]pyrene and 

14 phenanthrene. 

15 Cavalieri et al. (1991) treated Sprague-Dawley rats with single intramammillary 

16 injections of dibenzo[a,l]pyrene into the left mammary glands and followed them for up to 24 

17 weeks. Tumors of the mammary gland, mesenchymal tissue, or skin were recorded. 

18 Dibenzo[a,l]pyrene produced tumors in all animals at both doses. 

19 In six studies, tumors were not induced after exposure to any target PAH. 

20 Intramammillary injection of dibenz[a,h]anthracene and benz[a]anthracene did not induce 

21 mammary tumors in rats (Cavalieri et al., 1988b). Pregnant mice receiving subcutaneous 

22 injection of pyrene did not develop tumors, nor did their offspring (Nikonova, 1977). Rats 

23 treated either intravenously or intramuscularly with benz[a]anthracene did not develop either 

24 mammary or injection site tumors (Pataki and Huggins, 1969). Similarly, benz[a]anthracene was 

25 not tumorigenic after intramuscular injection in rats (Sugiyama, 1973) or buccal pouch painting 

26 in hamsters (Solt et al., 1987). Finally, benzo[e]pyrene was not tumorigenic when it was 

27 implanted into tracheas transplanted subcutaneously into isogenic rats (Topping et al., 1981). 

28 Among the tumor bioassays that used alternative exposure routes in Table 4-5, four 

29 studies met the selection criteria for use in this analysis.

30 

31 4.3.2. In Vivo Studies of Cancer-Related Endpoints

32 The database of cancer-related endpoints measured after in vivo exposure to PAHs is 

33 much smaller than the in vitro database. Endpoints examined after in vivo exposure include 

34 mutagenicity, DNA adducts, and clastogenicity or sister chromatid exchange. As with the in 

35 vitro database, only studies of selected PAHs that included benzo[a]pyrene as a reference 

36 compound were reviewed. Each study that was reviewed for consideration in relative potency 

37 development is presented in tabular format in subsequent sections. The tables summarize study38 specific information and indicate whether a particular study is considered useful for dose-
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1 response assessment. The text provides an overall description of the available studies, including 

2 a general description of the methodology used for each study type, the results, and the 

3 weaknesses or problems associated with specific studies or study types. 

4 

5 4.3.2.1. DNA Adducts

6 Eighteen studies evaluating DNA adduct formation for PAHs and benzo[a]pyrene were 

7 identified in the database (Table 4-6). Nine studies presented quantitative data for DNA adduct 

8 formation and are discussed below. Among studies with data potentially useful for RPF 

9 derivation, the route of exposure was intramammillary injection in one study (Arif et al., 1997), 

10 intraperitoneal injection in five studies (Kligerman et al., 2002; Nesnow et al., 1998a, 1996, 

11 1995; Ross et al., 1995; Mass et al., 1993), dermal in three studies (Hughes and Phillips, 1990; 

12 Cavalieri et al., 1981b; Phillips et al., 1979), and oral in one study (Kligerman et al., 2002). 

Adducts were identified by [32 13 P]-postlabeling in all of the studies except for two by Phillips et al. 

(1979) and Cavalieri et al. (1981b), which utilized [3H]- or [14 14 C]-radiolabeled PAHs. Two 

15 papers described experiments with a single time point(s) at 24 or 48 hours (Arif et al., 1997, 

16 Hughes and Phillips, 1990), whereas the rest had multiple time points. The duration of exposure 

17 was as short as 4 hours (Cavalieri et al., 1981b), although 24 hours was usually the first time 

18 point(s) in time course studies. The longest duration for a time course study was 84 days 

19 (Hughes and Phillips, 1990), but most were 3 weeks or less. The tissues evaluated included 

20 mammary epithelium (Arif et al., 1997), skin (Hughes and Phillips, 1990; Cavalieri et al., 1981b; 

21 Phillips et al., 1979), liver and peripheral blood lymphocytes (Kligerman et al., 2002; Nesnow et 

22 al., 1993b), and lung (Nesnow et al., 1998a, 1993b; Arif et al., 1997; Ross et al., 1995; Mass et 

23 al., 1993; Hughes and Phillips, 1990). 

24 Dermal-exposure studies typically involved application of the chemical in solution to the 

25 shaved dorsal skin of mice (Hughes and Phillips, 1990; Cavalieri et al., 1981b; Phillips et al., 

26 1979). After the scheduled sacrifice, the treated skin was excised and frozen; a scalpel was used 

27 to scrape away the dermis from the epidermis that was subsequently powdered in liquid nitrogen. 

28 In one study, the lung was also excised and frozen in liquid nitrogen (Hughes and Phillips, 

29 1990). DNA was isolated from the frozen epidermis or lung. Liquid scintillation counting was 

used to quantify DNA adducts to PAH labeled with [3H] or [14 30 C] (Cavalieri et al., 1981b; Phillips 

et al., 1979). For [32 31 P]-postlabeling, DNA was treated to selectively dephosphorylated 

32 nonadducted nucleotides; after postlabeling, adducts were resolved by sequential anion-exchange 

33 thin layer chromatography on PEI-cellulose plates in several directions using three solvents 

34 (Hughes and Phillips, 1990). Adduct spots on chromatograms were located by autoradiography, 

35 after which the spots were excised and radioactivity levels were determined by Cerenkov 

36 counting. 
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1 Compounds administered by intraperitoneal or intramammillary injection were also 

2 delivered in solution. As in dermal-exposure studies, DNA was isolated from frozen tissues and 

adducts were identified by [32 3 P]-postlabeling and quantified via autoradiography.

4 Most studies reported the mean number of adducts formed within a tissue per unit of 

5 DNA, with time-course data displayed graphically. Peak values were sometimes called out 

6 specifically in the text or tables. As the shapes of dose-response curves differ among different 

7 PAHs, the peak value is an imprecise measure for comparing the relative adduct-forming 

8 potency of the different compounds. The TIDAL has also been used for reporting results for a 

9 time-course study (Ross et al., 1995). The TIDAL value is the area under the curve (AUC) for 

10 adduct persistence (based on the rate of adduct formation and repair) for the duration of the 

11 study. The TIDAL value expresses the total DNA adduct burden experienced by the tissue from 

12 the time of treatment to the end of the study. The TIDAL versus administered dose curve 

13 provides a convenient way to compare adduct-forming potency for different PAHs in time14 course experiments. An important limitation of the TIDAL approach is the inherent assumption 

15 that the ratios of specific adducts are relatively constant across dose and time course. Ross et al. 

16 (1995) demonstrated that this assumption was valid for several different PAHs; however, it was 

17 also noted that two adducts of benzo[a]pyrene in rat liver did not conform to this general pattern. 

18 Ross et al. (1995) presented data for lung adenoma incidence (measured at 8 months) in 

19 several ways: as a function of administered dose, as a function of adduct levels per dose 

20 measured 24 hours after dosing (results for 3 days postdosing were mentioned but not shown), as 

21 a function of TIDAL values measured over 21 days (during which period adduct levels were 

22 specifically quantified), and as a function of TIDAL values extrapolated to 8 months. The 

23 relative tumor induction potencies of the studied PAHs were similar for each assay for a single 

24 PAH when described as functions of administered dose, the adduct levels per dose at 3 days, the 

25 TIDAL values over 21 days, or the TIDAL values extrapolated to 8 months. The relative 

26 potencies for tumor incidence as a function of adduct levels at 24 hours were not similar to those 

27 associated with the other measures of exposure. Ross et al. (1995) suggested that 

28 pharmacokinetic differences in adduct formation among the PAHs were responsible for the 

29 discrepancy, but suggested that peak levels could be used to compare the potencies of different 

30 PAHs if adduct formation for those PAHs followed similar kinetics. 

31 DNA adduct experiments were carried out in replicate and were usually analyzed 

32 statistically. It should be noted that, based on the work of Ross et al. (1995), relative potencies 

33 determined from studies that administered a single dose level and measured adducts at a single 

34 time point will be less reliable unless the shapes of the adduct formation curves are similar. 

35 However, the single dose and single measurement studies were also used for dose-response 

36 assessment. 

37 Among the in vivo DNA adduct studies shown in Table 4-6, nine studies met the 

38 selection criteria for use in this analysis.
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1 

2 4.3.2.2. Clastogenicity or Sister Chromatid Exchange Frequency 

3 The database included 13 studies in which clastogenic effects or frequency of sister 

4 chromatid exchanges of benzo[a]pyrene and at least one other PAH were tested in whole animal 

5 systems. Table 4-7 lists the studies along with important study design details. The clastogenic 

6 endpoints measured in these studies were micronuclei, chromosome gaps and breaks, and 

7 nonspecific aberrations; sister chromatid exchanges were also measured. These studies were all 

8 conducted in rodents, including mice, rats, and hamsters. 

9 Eight of the studies evaluated micronuclei, sister chromatid exchanges, or chromosome 

10 gaps or breaks in bone marrow from treated mice or hamsters (Allen et al., 1999; Katz et al., 

11 1981; Paika et al., 1981; Salamone et al., 1981; Tsuchimoto and Matter, 1981; Roszinsky-Kocher 

12 et al., 1979; Bayer, 1978; Sugiyama, 1973). In these studies, one or two doses of PAH were 

13 injected intraperitoneally into the animals, and sacrifice occurred at various time points thereafter 

14 (typically 24 hours after). Bone marrow smears were examined microscopically and scored for 

15 micronuclei, sister chromatid exchanges, gaps, or breaks. 

16 He and Baker (1991) applied multiple dose levels of chrysene or phenanthrene to the skin 

17 of hairless mice and harvested keratinocytes upon sacrifice 24 hours later. The keratinocytes 

18 were incubated for 2 days and treated with cytochalasin B to identify binucleated cells. After 

19 4 days in vitro, cells were mounted on slides and examined microscopically for micronuclei. 

20 Results were reported as the percent of binucleated cells with one or more micronuclei among 

21 the total number of binucleated cells scored. Chrysene treatment resulted in a dose-related 

22 increase in micronuclei, while pyrene did not. 

23 Kligerman et al. (2002, 1986) measured sister chromatid exchanges and/or micronuclei in 

24 the blood of mice or rats given a single dose of PAH either orally or intraperitoneally. The study 

25 by Oshiro et al. (1992) involved two or four oral doses of pyrene or anthracene in mice. Blood 

26 obtained from the tail 24 hours after the last treatment was examined microscopically and 

27 micronuclei were scored in polychromatic erythrocytes. In an unusual study design, Sirianni and 

28 Huang (1978) measured sister chromatid exchanges in V79 cells placed in a diffusion chamber 

29 implanted in the peritoneal cavity of mice. 

30 Thirteen individual PAHs were evaluated in these studies. Only chrysene gave positive 

31 results for more than one endpoint (for sister chromatid exchange and micronucleus frequency; 

32 He and Baker, 1991; Roszinsky-Kocher et al., 1979). Five other PAHs (phenanthrene, 

33 dibenz[a,h]anthracene, benz[a]anthracene, benzo[b]fluoranthene, and benzo[e]pyrene) increased 

34 the frequency of sister chromatid exchange in hamster bone marrow after intraperitoneal 

35 administration (Roszinsky-Kocher et al., 1979). Bayer (1978) also reported an increase in sister 

36 chromatid exchange frequency in hamster bone marrow after phenanthrene administration (high 

37 dose only). Anthracene and pyrene consistently gave negative results in several studies (Oshiro 

38 et al., 1992; He and Baker, 1991; Katz et al., 1981; Paika et al., 1981; Salamone et al., 1981; 
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1 Tsuchimoto and Matter, 1981; Roszinsky-Kocher et al., 1979; Sirianni and Huang, 1978). 

2 Dibenzo[a,i]pyrene and benzo[g,h,i]perylene each gave negative results in an assay for bone 

3 marrow micronuclei (Katz et al., 1981). 

4 Among studies with positive results, only He and Baker (1991), Kligerman et al. (1986), 

5 and Bayer (1978) administered PAHs at multiple dose levels. Bayer (1978) observed a positive 

6 response only with the highest dose of phenanthrene. Of the single dose studies, only 

7 Roszinsky-Kocher et al. (1979) reported responses clearly differing from controls. 

8 Among the in vivo clastogenicity or sister chromatid exchange studies shown in 

9 Table 4-7, 10 studies met the selection criteria for use in this analysis.

10 

11 4.3.2.3. In Vivo Mutagenicity 

12 The PAH database contains several studies that evaluate specific mutagenic end points 

13 following in vivo exposure to PAHs (see Table 4-8). These studies include mutagenicity 

14 experiments in Drosophila melanogaster, an intraperitoneal host-mediated assay using 

15 Salmonella strains or yeast, and DNA sequence analysis of specific codons in the Ki-ras 

16 oncogene in mouse lung tumors. 

17 Most Drosophila studies administered PAH compounds to either the suspension media or 

18 to the diet for 48–72 hours prior to cross-mating and analysis of mutations (Frolich and Wurgler, 

19 1990; Valencia and Houtchens, 1981; Fahmy and Fahmy, 1980). One study used abdominal 

20 injection as an exposure pathway (Zijlstra and Vogel, 1984). The mutagenic endpoints evaluated 

21 included somatic mutations (i.e., eye color mosaicism, wing spots) (Frolich and Wurgler, 1990; 

22 Fahmy and Fahmy, 1980) or sex-linked recessive lethal mutations (Zijlstra and Vogel, 1984; 

23 Valencia and Houtchens, 1981). Only two PAHs were evaluated in the Drosophila studies in 

24 addition to benzo[a]pyrene (benz[a]anthracene and pyrene), and the results were either negative 

25 or inconsistent in all studies (Frolich and Wurgler, 1990; Zijlstra and Vogel, 1984; Valencia and 

26 Houtchens, 1981; Fahmy and Fahmy, 1980). A significant effect was seen for benz[a]anthracene 

27 only with cross-breeding of strains selected for enhanced metabolic activity (Frolich and 

28 Wurgler, 1990). No effect was observed using the standard strains. 

29 An intraperitoneal host-mediated assay was described by Simmon et al. (1979). Five 

30 PAHs (anthracene, benz[a]anthracene, benzo[e]pyrene, chrysene, and phenanthrene) were 

31 administered to Swiss Webster mice by gavage or intramuscular injection (single dose only). 

32 Microorganisms (S. typhimurium and Saccharomyces cerevisiae) were injected intraperitoneally 

33 into exposed mice and were recovered 4 hours later for mutation analysis. Negative results were 

34 observed and the host-mediated assay system was considered insensitive for detecting 

35 carcinogenic PAHs. 

36 A series of studies have investigated the mutation sequence in codons 12 and 61 of the 

37 Ki-ras oncogene from PAH-induced lung adenomas in A/J mice (Nesnow et al., 1998a, 1996, 

38 1995; Mass et al., 1993). As discussed in Section 2.4 (Similarities in Mode of Action for PAHs), 
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1 the purpose of these studies was to correlate the tumorigenic potency of specific PAHs with the 

2 formation of DNA adducts and the mutation of specific codons in the Ki-ras oncogene. Six non3 alkylated PAHs were utilized in these studies (benzo[a]pyrene, benz[j]aceanthrylene, 

4 benzo[b]fluoranthene, dibenz[a,h]anthracene, cyclopenta[c,d]pyrene, and dibenzo[a,l]pyrene). 

5 Mutation analysis of the Ki-ras oncogene at codons 12 and 61 was carried out in PAH-induced 

6 lung adenomas using polymerase chain reaction (PCR) amplification and dideoxy nucleotide 

7 sequencing methods. The primary mutation type for benzo[a]pyrene, benzo[b]fluoranthene, and 

8 dibenzo[a,l]pyrene was the GGT→TGT mutation. This guanine mutation was correlated with 

9 the formation of diol epoxide guanine adducts. The GGT→CGT mutation was the primary 

10 mutation type for benz[j]aceanthrylene and cyclopenta[c,d]pyrene. The CGT mutation was 

11 associated with the formation of cyclopenta-guanine adducts and increased tumorigenic potency 

12 (i.e., >90 adenomas per mouse) in A/J mice. Dibenz[a,h]anthracene was the only PAH evaluated 

13 that did not induce mutations in Ki-ras codons 12 or 61. This compound produced diol epoxide 

14 guanine adducts and lung adenomas in A/J mice, suggesting a possible interaction at a different 

15 genetic target. The Ki-ras mutation analysis data were presented as percent of tumors with a 

16 specific mutation at either codon 12 or 61. No dose-response data were provided. 

17 Among the in vivo mutagenicity studies shown in Table 4-8, only one study met the 

18 selection criteria for use in this analysis.

19 

20 4.3.3. In Vitro Studies of Cancer-Related Endpoints 

21 Many in vitro studies of cancer-related endpoints are present in the PAH database. As 

22 previously discussed, only those studies that included at least one selected PAH and 

23 benzo[a]pyrene as a reference compound were reviewed. Each study that was reviewed for the 

24 purpose of RPF development is included in Tables 4-9 through 4-14. The tables summarize 

25 study-specific information and indicate whether a particular study is considered useful for dose26 response assessment. The text provides an overall description of the available studies, including 

27 a general description of the methodology used for each study type, the results, and the 

28 weaknesses or problems associated with specific studies or study types. 

29 

30 4.3.3.1. Bacterial Mutagenicity 

31 The bacterial mutagenicity of many PAHs has been extensively studied (39 studies with 

32 benzo[a]pyrene; see Table 4-9). All of the studies used the Ames assay in S. typhimurium. A 

33 total of 38 PAHs have been evaluated for their ability to induce mutations in bacterial systems. 

34 The Ames Salmonella assay is a bacterial reverse mutation assay, which measures the 

35 frequency at which histidine-independent bacteria arise from histidine-requiring bacterial strains 

36 in the presence of a chemical mutagen. The results are generally expressed as either the number 

37 of revertant colonies per plate or the number of revertants/nmol of the test compound (calculated 

38 from the linear portion of the dose-response curve). Several strains of S. typhimurium have been 
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1 used to evaluate specific PAH mutation types; for example TA98, TA1537, and TA1538 detect 

2 various frameshift mutations, TA1535 responds to base-pair substitution and TA100 responds to 

3 a broad spectrum of mutations. Metabolism to reactive intermediates is required for PAH 

4 mutagenicity in Salmonella and many metabolic activation systems have been employed. Rat 

5 liver postmitochondrial supernatant (known as S9) from Aroclor-induced rats is most often used, 

6 although other rodent species and enzyme inducers are sometimes employed. Isolated rat 

7 hepatocytes or purified mixed-function oxidase enzymes were occasionally utilized for metabolic 

8 activation of PAHs. 

9 Of the PAHs tested for bacterial mutagenicity, most were considered positive in at least 

10 one study under optimal study conditions. Compounds that produced negative results in multiple 

11 studies include anthracene, fluorene, phenanthrene, and pyrene. The primary weakness of the 

12 bacterial mutagenicity database for PAHs is the limited amount of multiple-dose data for many 

13 PAHs. Many studies report findings at a single dose level for several PAHs. 

14 Among the in vitro bacterial mutagenicity studies shown in Table 4-9, 29 studies met the 

15 selection criteria for use in this analysis.

16 

17 4.3.3.2. Mammalian Mutagenicity 

18 Studies that evaluate the mutagenicity of target PAHs in mammalian cells are described 

19 in Table 4-10 (29 studies). The most common cell types used in these studies were the 

20 V79 Chinese hamster cells and the L5178Y mouse lymphoma cells. Other cell types include 

21 human epidermal keratinocytes, TK6 human lymphoblasts, human epithelial cells (HS1 HeLa), 

22 human foreskin fibroblasts (D-550), mouse fibroblasts, rat embryo cells, rat liver epithelial cells 

23 (ARL-18), and Chinese hamster ovary (CHO) cells. A total of 14 PAHs have been evaluated for 

24 their ability to induce mutations in mammalian cell systems. 

25 Each of the mammalian cell assays detects forward mutations that confer resistance to a 

26 toxic chemical. Mutations in the hypoxanthine-guanine phosphoribosyl transferase gene (HPRT) 

27 result in resistance to purine analogs such as 6-thioguanine, 8-azaguanine, and ouabain. HPRT 

28 mutations induced by PAHs were most often measured in V79 Chinese hamster cells, but have 

29 also been detected in human, rat, and mouse cell lines. Forward mutation at the thymidine 

30 kinase locus (TK) is measured as colony growth in the presence of thymidine analogs (e.g., 

31 trifluorothymidine or 5-bromo-2’-deoxyuridine). PAH-induced TK mutations were measured in 

32 mouse lymphoma cells (L5178Y) and human lymphoblasts. Forward mutation assays are 

33 considered to respond to a variety of mutation types (including frameshift, base-pair substitution, 

34 deletions and rearrangements or complex mutations). Exogenous metabolic activation is 

35 required for PAH mutagenicity in most mammalian cell assays. This was accomplished using a 

36 rat liver S9 mix or cocultivation with other rodent cells able to metabolize PAHs to reactive 

37 intermediates (i.e., hamster embryo cells, fibroblasts, or hepatocytes; rat hepatocytes). The 
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1 results of forward mutation assays in mammalian cell lines are generally expressed as mutant 

frequency/10x2 survivors. 

3 Of the 26 PAHs tested for mammalian cell mutagenicity, all were considered positive in 

4 at least one study under optimal study conditions. Compounds that produced negative results in 

5 some studies include anthracene, benzo[e]pyrene, phenanthrene, and pyrene. Benzo[a]-

6 anthracene produced positive findings in seven studies and negative findings in four studies. The 

7 mammalian mutagenicity studies generally provide more multi-dose data than the bacterial 

8 mutagenicity studies. 

9 Among the in vitro mammalian mutagenicity studies shown in Table 4-10, 27 studies met 

10 the selection criteria for use in this analysis.

11 

12 4.3.3.3. Morphological/Malignant Cell Transformation 

13 Twenty-five studies examined the capacity of benzo[a]pyrene and other PAHs to 

14 transform cells in culture (Table 4-11). All of these studies were conducted using mammalian 

15 cells, most commonly mouse or hamster embryo cells. A few studies added feeder cells or rat 

16 liver homogenate to enhance metabolic activation in the test system; however, the majority relied 

17 on the intrinsic metabolic capacity of the cells. The general test protocol involved seeding the 

18 cultured cells in Petri dishes followed by exposure to a solution of the test compound, usually for 

19 a period of 24 hours. The cells were then cultured for about 6 weeks before being fixed and 

20 stained. Transformed colonies (foci) were scored based on characteristics such as cell piling, 

21 criss-crossing, basophilic staining, and/or invasion of surrounding (nontransformed) cell 

22 monolayer. In studies conducted by some laboratories, foci were classified as Type II or 

23 Type III; the latter category included those with invasion of the surrounding monolayer, highly 

24 criss-crossed arrays, and deep staining. Data were generally reported as the number of foci 

25 (colony of transformed cells) per dish or per surviving cells and/or the percent of dishes with 

26 foci. 

27 In a few cases (e.g., Greb et al., 1980), transformation was assessed by growth of treated 

28 cells in soft agar. Transformed cell colonies growing in semi-solid agar are capable of 

29 anchorage-independent growth. 

30 Three studies (Evans and DiPaolo, 1975; Kakunaga, 1973; DiPaolo et al., 1972) 

31 confirmed the identification of malignant cells by injecting the transformed cells into rodents and 

32 following tumor induction in the animals. In all three cases, cells identified as transformed gave 

33 rise to tumors, while the cells without these characteristics did not. 

34 Cell transformation assays were identified that included 22 individual PAHs other than 

35 benzo[a]pyrene. Dibenz[a,h]anthracene consistently gave rise to transformed cells in all but one 

36 of the seven studies in which it was tested. Cyclopenta[c,d]pyrene, indeno[1,2,3-c,d]pyrene, 

37 benzo[j]aceanthralene, benz[e]aceanthrylene, and dibenz[k,mno]acephenanthrylene were each 

38 tested in a single study and gave positive results. Benz[a]anthracene, pyrene, phenanthrene, 
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1 benzo[e]pyrene, and anthracene each gave negative results in a number of studies, while 

2 fluoranthene, benzo[k]fluoranthene, dibenz[j,mno]acephenanthrylene, naphth[1,2,3-mno]ace3 phenanthrylene, and aceanthrylene were each tested in a single study and gave negative results. 

4 Only a single dose of the target PAH was applied in 8 of the 26 studies of in vitro morphological/

5 malignant cell transformation. 

6 Among the in vitro morphological/malignant transformation studies shown in Table 4-11, 

7 19 studies met the selection criteria for use in this analysis.

8 

9 4.3.3.4. DNA Adducts

10 Several studies (14) were identified in which DNA adducts were measured after either 

11 whole cells or extracted DNA were incubated with benzo[a]pyrene and at least one other PAH. 

12 Table 4-12 shows general study details for these studies. Most of the studies involved 

13 measurement of DNA adducts in whole mammalian cells, while some measured adducts formed 

14 when PAHs were incubated with extracted DNA. Whole cells were usually incubated with 

15 PAHs for about 24 hours, while extracted DNA was exposed to PAH solutions for a shorter time 

16 period (1–3 hours). Some of the studies added metabolic activation (usually rat liver 

17 microsomes) to the incubation solution. Melendez-Colon et al. (2000) evaluated DNA adduct 

18 formation after dibenzo[a,l]pyrene exposure in two cell types: one having significant CYP450 

19 activity (MCF-7 cells) and one lacking significant CYP450 activity (HL-60). The authors 

20 reported that adducts were formed in the cells having CYP450 activity, but no adducts were 

21 formed in the cells lacking such activity. 

Identification and quantification of adducts was generally done using a [32 22 P]-postlabeling 

23 assay as follows. After exposure, DNA was isolated and digested to mononucleotides. 

Mononucleotides were radiolabeled with [32 24 P]-ATP, separated with thin layer chromatography, 

25 and visualized by autoradiography. Relative adduct labeling was measured using a scintillation 

counter. A few early studies used [326 H]-labeled PAHs to identify and quantify adducts. In some 

27 cases, adducts were identified by high-performance liquid chromatography and GC-MS. 

28 The 14 studies reviewed examined 15 PAHs other than benzo[a]pyrene. Apart from 

29 phenanthrene, which did not result in measurable DNA adducts when incubated with calf thymus 

30 DNA under various conditions (Bryla and Weyand, 1992), each of the PAHs produced 

31 measurable DNA adducts in at least one study. 

32 Major limitations associated with some of the in vitro DNA adduct data for relative 

33 potency development include the lack of data at multiple PAH exposure levels, the use of 

34 extracted DNA rather than whole cell assays, and the inconsistent use of extrinsic metabolic 

35 activation sources. Only three studies with positive adduct findings reported adduct 

36 measurements at multiple doses (concentrations) of PAH (Binkova et al., 2000; Melendez-Colon, 

37 2000; Bryla and Weyand, 1992). Three studies used extracted DNA rather than whole cells to 

38 measure DNA binding (Segerback and Vodicka, 1993; Bryla and Weyand, 1992; Grover and 
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1 Sims, 1968). Finally, the available studies on DNA adduct formation use cell types with varying 

2 degrees of PAH metabolic capacity, with and without added metabolic activation sources. Both 

3 the types and the quantities of DNA adducts formed are likely to depend on the level of 

4 metabolic activation for most PAHs. 

5 Among the in vitro DNA adduct studies shown in Table 4-12, 10 studies met the 

6 selection criteria for use in this analysis.

7 

8 4.3.3.5. DNA Damage/Repair 

9 Twenty-four reports in the database evaluated the effects of one or more PAHs on DNA 

10 damage, repair, or synthesis. Table 4-13 summarizes the study design information and results of 

11 these studies. Studies included measures of unscheduled DNA synthesis and DNA damage. 

Unscheduled DNA synthesis was generally measured by increased radiolabeled (312 H) thymidine 

13 uptake in treated cells versus untreated cells. DNA damage was measured either using the 

14 alkaline elution assay for DNA strand breakage in mammalian cells, or using the differential 

15 killing of DNA repair-deficient bacterial strains. Metabolic activation of PAHs was most often 

16 accomplished using a rat liver S9 mix. 

17 Twenty-eight different PAHs have been tested for effects on DNA in one or more assays. 

18 In general, pyrene, anthracene, phenanthrene, perylene, fluorene, and benzo[e]pyrene gave 

19 negative results in multiple studies. Chrysene gave negative results in four assays and positive 

20 results in one assay (Mersch-Sundermann et al., 1992). More positive than negative results were 

21 reported for benz[a]anthracene, dibenz[a,h]anthracene, and dibenz[a,c]anthracene. Other PAHs 

22 were tested only once, or gave roughly an equal frequency of positive and negative responses in 

23 these assays. 

24 Although a large number of PAHs have been tested for DNA damage/repair, the database 

25 includes both bacterial and mammalian cells and several different genotoxic endpoints. In 

26 addition, the use of external metabolic activation, or cell types with intrinsic metabolic capacity, 

27 was inconsistent across these studies. These limitations make it difficult to compare studies 

28 using the same target PAHs. 

29 Among the in vitro DNA damage/repair studies shown in Table 4-13, 15 studies met the 

30 selection criteria for use in this analysis.

31 

32 4.3.3.6. Clastogenicity or Sister Chromatid Exchange Frequency 

33 The database contains 18 studies in which clastogenicity or sister chromatid exchange 

34 frequency was measured in cultured cells after exposure to benzo[a]pyrene and at least one other 

35 PAH (Table 4-14). A wide variety of cell types was used in these assays, including hamster 

36 liver, lung, CHO and V79 cells; rat liver epithelial cells; human teratocarcinoma epithelial cells; 

37 rat and human mammary epithelial cells; mouse, rat, and human fibroblasts; human 

38 lymphocytes; and guinea pig fetal cells. A number of the studies used a metabolic activation 
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1 system, typically either rat liver S9 or coculture with a cell type able to metabolize PAHs. While 

2 laboratory methods varied widely, the general approach involved treating the cultured cells with 

3 a solution of the test compound, either with or without metabolic activation. Usually, 

4 bromodeoxyuridine was added to the growth medium to provide a means of staining metaphase 

5 chromosomes, and colcemid was used to arrest mitotic cells. Chromosomes were examined 

6 microscopically and aberrations or exchanges were scored visually. In most cases, the endpoint 

7 examined was frequency of sister chromatid exchanges. Other endpoints included frequency of 

8 micronuclei and scoring of chromosomal aberrations such as breaks, gaps, deletions, etc. 

9 Only eight PAHs (anthracene, benz[a]anthracene, benzo[e]pyrene, cyclopenta10 [c,d]pyrene, fluoranthene, perylene, phenanthrene, and pyrene) have been tested for clastogenic 

11 effects in vitro. In many cases, the available studies were aimed at evaluating the validity of a 

12 given test system to predict carcinogenicity. In these studies, a range of compounds of known or 

13 believed carcinogenicity were used. Often, benzo[a]pyrene was included as a known carcinogen, 

14 and other PAHs were chosen because they were known or believed to be noncarcinogenic or 

15 weakly carcinogenic. 

16 Among the tested compounds, four gave positive results in at least one study. With few 

17 exceptions, PAHs administered without metabolic activation gave negative responses in these 

18 assays. Cyclopenta[c,d]pyrene was reported to increase the frequency of sister chromatid 

19 exchanges in two assays, one with and one without metabolic activation (Murison, 1988; 

20 Krolewski et al., 1986). Benz[a]anthracene gave positive results in three studies of sister 

21 chromatid exchange induction (Mane et al., 1990; Tong et al., 1983; 1981a) and negative results 

22 in a fourth (Warshawsky et al., 1995). Kochhar (1982) reported a dose-dependent increase in 

23 chromosomal aberrations in V79 cells treated with benz[a]anthracene in the absence of metabolic 

24 activation. Perylene increased aberrations in one system (Popescu et al., 1977), but did not 

25 increase sister chromatid exchanges in another (Sirianni and Huang, 1978). Likewise, pyrene 

26 gave positive results in a number of studies that included metabolic activation (Evans and 

27 Mitchell, 1981; Perry and Thomson, 1981; Popescu et al., 1977) and negative results in several 

28 that did not include activation (DeSalvia et al., 1988; Tong et al., 1983, 1981a; Dean, 1981; Abe 

29 and Sasaki, 1977). 

30 The clastogenicity and sister chromatid exchange data for PAHs are variable with respect 

31 to cell type and use of extrinsic metabolic activation. Some cells have intrinsic metabolic 

32 activity, while others require activation from an external source. The degree to which metabolic 

33 activation is required for PAHs to exert a clastogenic effect in cell cultures is not well 

34 established. Another limitation of these data stems from the fact that a small number of PAHs, 

35 many traditionally believed to be noncarcinogenic or weakly carcinogenic, have been tested for 

36 clastogenic effects in vitro. 

37 Among the in vitro clastogenicity/sister chromatid exchange studies shown in Table 4-14, 

38 10 studies met the selection criteria for use in this analysis.
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1 

2 4.4. SUMMARY OF INFORMATION AVAILABLE TO DEVELOP RPFS FOR 

3 INDIVIDUAL PAHs 

4 The PAH database contains several different types of data that may be used to estimate 

5 relative potencies of individual PAHs. The data were summarized in Section 4.3 and include in 

6 vivo tumor bioassays using various routes of exposure and data for cancer-related endpoints 

7 from both in vivo and in vitro studies. As discussed above, the concurrent testing of 

8 benzo[a]pyrene as a reference compound was considered essential to allow for RPF calculation. 

9 The introduction to Section 4.3 lists criteria for selecting studies or data sets for use in the 

10 analysis. Studies that met these criteria were used in the development of the RPF approach. 

11 Section 5 discusses methods used for dose response assessment and RPF calculation from each 

12 study or dataset, and Section 6 discusses the selection of PAHs to be included in the RPF 

13 approach using a weight of evidence evaluation of the available data. Section 7 describes the 

14 derivation of final RPFs for each PAH included in the analysis. 

15 

16 

17 5. METHODS FOR DOSE-RESPONSE ASSESSMENT AND RPF CALCULATION 

18 

19 

20 A discussion of the available data on PAH carcinogenicity and cancer-related endpoints 

21 and criteria for selection of studies was presented in Section 4. This section describes the 

22 selection of dose-response data and methods for dose-response assessment and RPF calculation 

23 from the selected datasets. The dose-response data extracted from each study with positive 

24 results and the results of the statistical analyses are shown in Appendix C. Appendix C also 

25 contains information regarding the source of the dose-response data (i.e., the figure or table 

26 number from the study and the particular data points that were used in the dose-response 

27 assessment) and additional comments on the use of the data for dose-response assessment and 

28 RPF calculation. The results of the RPF calculations are shown in tables in Appendix E. These 

29 tables provide summary information for each study, including the PAHs that were tested, the 

30 data used to estimate the slopes (point estimate or BMD model result), the calculated RPF value, 

31 and any specific comments related to the data analysis. 

32 

33 5.1. CHOICE OF DOSE-RESPONSE DATA 

34 For each of the endpoints evaluated in Section 4 (dermal, intraperitoneal, subcutaneous, 

35 and other route bioassays; in vivo DNA adducts; in vivo clastogenicity or sister chromatid 

36 exchange frequency; in vitro bacterial and mammalian mutagenicity; in vitro morphological/

37 malignant transformation; in vitro clastogenicity or sister chromatid exchange frequency; and 

38 other in vitro endpoints [DNA adducts, unscheduled DNA synthesis, DNA damage, etc.]) there 
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1 was at least one study that met selection criteria. For those studies with positive findings, dose2 response data were extracted for dose-response assessment and calculation of RPFs. Data that 

3 were reported in graphical format in published studies were digitized (Grab It!TM Graph 

4 Digitizer, Datatrend Software) to identify the dose-response data points. 

5 As discussed in Section 4.3, statistics were used for tumor bioassay data to determine 

6 whether the tumor incidence or multiplicity observed at a particular dose represented a 

7 statistically significant increase over controls. If statistical analyses were not described in the 

8 original report, incidence data were analyzed using Fisher’s Exact test and the Cochran-Armitage 

9 trend test. Positive findings were indicated by a significant (p < 0.05) difference for at least one 

10 dose group by comparison to control (in Fisher’s Exact or an equivalent test) or a significant 

11 dose-response trend (Cochran-Armitage or equivalent) for multi-dose studies. For tumor 

12 bioassay data reported as tumor count, a t-test was conducted (when variance data were 

13 available) to determine whether the count was significantly different from control (p < 0.05). 

14 The results of the statistical analyses are shown with the dose-response data in Appendix C. 

15 The tumor bioassays that reported both incidence and tumor count were unique in 

16 offering two different datasets for the same study. For each dose of each PAH in the tumor 

17 bioassays, the decision to calculate an RPF, and in some instances, the selection of the point of 

18 departure, was based on whether the tumor incidence or count was statistically significantly 

19 increased over the control; if there was a significant increase, an RPF was calculated. There 

20 were a few instances where the statistical tests for tumor incidence and tumor number were 

21 inconsistent (i.e., the incidence of tumors was statistically significantly increased, but the tumor 

22 count was not, or vice versa). Sometimes, this circumstance existed only at a low dose, with 

23 consistent findings at higher doses, so the conclusion as to whether there was treatment-related 

24 tumorigenicity (and whether an RPF should be calculated) was clear. In one case, however, the 

25 conclusion as to whether there was treatment-related tumorigenicity was not clear. In female 

26 mice exposed at the high dose of fluoranthene in the study reported by Busby et al. (1984), the 

27 lung tumor count was significantly increased (albeit borderline, p = 0.0343) while the incidence 

28 was not, and neither was statistically significantly increased at the lower dose. For the purpose 

29 of this analysis, the multiplicity data were treated as an independent measure of carcinogenic 

30 potency, and an RPF was calculated for the statistically increased tumor count irrespective of the 

31 analysis of incidence. It should be noted that average tumor count can be skewed by an unusual 

32 response in a single animal, and no information was available to determine whether such 

33 response represented an anomaly unrelated to exposure or an unusual susceptibility to the 

34 exposure. Thus, reliance on statistical analysis of mean tumor count alone as a measure of 

35 carcinogenic response may be subject to additional uncertainty. 

36 For cancer-related endpoint data, each study authors’ conclusions regarding a positive or 

37 negative response for each PAH were accepted, and RPFs were calculated when positive results 

38 were reported. 
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1 In a few cases, the only data in a given publication were given as relative potency 

2 (relative to benzo[a]pyrene). For these publications, which included only in vitro cancer-related 

3 endpoint data (primarily mutagenicity), the relative potency estimates calculated by the authors 

4 were used without modification (except for dose adjustment where appropriate; see Section 5.5). 

5 

6 5.2. OVERALL FORM OF RPF ESTIMATE 

7 The overall goal of the dose-response analysis was to calculate ratios representing the 

8 relative potency of a given PAH compared with benzo[a]pyrene (i.e., RPFs). For all datasets, the 

9 RPF was defined as the ratio (PAHi:BaP) of the slopes of the dose-response curves in the low10 dose region, following the equation (eq 5-1) below:

11 

12 RPF = slope PAHi÷slope BaP (5-1) 

13 

14 Data available for calculation of RPFs consisted of both quantal and continuous 

15 endpoints. Quantal endpoints included tumor incidence or incidence of cancer-related endpoints 

16 (including frequency of mutations). Continuous endpoint datasets included tumor counts 

17 (number of tumors per animal) or cancer-related endpoints of a continuous-variable nature (e.g., 

18 number of sister chromatid exchanges, number of morphologically transformed colonies). Dose19 response assessment methods were specific to each type of endpoint (quantal or continuous) and 

20 differed depending on whether there were multiple dose groups or a single dose group in the 

21 dataset. Methods for multidose and single dose quantal and continuous data are described below. 

22 

23 5.3. RPF CALCULATION FOR MULTIDOSE DATASETS 

24 Dose-response modeling using U.S. EPA’s Benchmark Dose (BMD) Software (Version 

25 1.3.2) was conducted on multiple-dose data sets to estimate potency for both the target PAHs and 

26 benzo[a]pyrene. Modeled estimates consider information about the shape of the dose-response 

27 curve and are thus preferred over using a single dose group as the point of departure. 

28 Dose-response modeling. For multidose quantal data, the multistage model was used and 

29 the degree of the polynomial was assumed to equal the number of dose groups minus 2 (extra 

30 risk with background subtracted). The multistage model was selected because it is the preferred 

31 model for cancer risk assessment of animal bioassay data, and it provided a consistent model 

32 form for all of the datasets. For multidose continuous data, the linear model was selected for all 

33 datasets, as it is the simplest model form for continuous data. For both quantal and continuous 

34 datasets, the goodness-of-fit criteria were used to evaluate model fit. If the model did not 

35 provide adequate fit to the data, high-dose groups were sequentially eliminated in an effort to 

36 achieve adequate fit. The focus of the modeling effort is on the low dose and response region, so 

37 doses and responses much higher than the benchmark response (BMR) are not as informative 

38 and can be eliminated to improve model fit. If dose-group elimination did not improve the 
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1 model fit, a point-estimate ratio approach was used (see Section 5.4). The BMD modeling 

2 output for all datasets that were successfully modeled are shown in Appendix D. 

3 Selection of BMR: Multidose data for both PAH and benzo[a]pyrene. For tumor 

4 incidence data, the BMR used in estimating the point of departure was a 10% increase in tumor 

5 incidence over controls (extra risk form). For cancer-related endpoints such as frequency of 

6 mutations, endpoint-specific points of departure were selected based on the background/control 

7 frequency of the endpoint and the detection limit of the assay. For example, a 1% frequency was 

8 selected for a control mutation frequency of 1/10,000 and a detection limit of two- to threefold 

9 above background. 

10 For multidose continuous data, the BMR used in estimating the point of departure was a 

11 change of 1 standard-deviation (1 SD) from the control mean. In the event that multiple-dose 

12 continuous data were reported in the absence of SD values, a point estimate ratio approach was 

13 employed to calculate the slope (see Section 5.4). 

14 Selection of BMR: Multidose data for PAH, single dose benzo[a]pyrene. Some studies 

15 included only one dose of benzo[a]pyrene as a positive control, while providing multiple-dose 

16 data for a selected PAH. In these cases, dose-response modeling was performed for the selected 

17 PAH and the BMR used for modeling was the observed response for benzo[a]pyrene adjusted for 

18 background response. For tumor incidence data, for example, if the benzo[a]pyrene dose was 

19 associated with a 60% extra risk for tumors, the BMR chosen for modeling the data for the PAH 

20 was 60% extra risk. RPFs were then calculated using a ratio of the slope factors calculated with 

21 equivalent points of departure (e.g., ED60). The goal of this approach was to compare PAH 

22 potencies at similar response locations on the dose-response curve. There is uncertainty 

23 associated with relative potency estimates calculated at the high end of the dose-response curves 

24 and using the resultant RPF for low-exposure scenarios, because the relative potency relationship 

25 between any two PAHs may be different at the low end, compared with the high end, of the 

26 dose-response curves. The uncertainties and limitations associated with the use of high-dose 

27 data to estimate relative potency are further discussed in Section 7. Data sets for which tumor 

28 incidence was ≥90% in the lowest dose group were not used to calculate potency estimates and 

29 RPFs, because the response is near plateau and such data provide insufficient information on the 

30 slope of the dose-response relationship. 

31 For continuous data, when a point estimate was used to estimate the slope for 

32 benzo[a]pyrene and modeling was used to estimate the slope for a given PAH, the BMR used for 

33 BMD modeling was a point value set at the response (e.g., mean number of tumors per animal 

34 for tumor multiplicity data) observed in the benzo[a]pyrene group, adjusted for response in the 

35 control group. This approach is consistent with the BMR used for quantal data when only a 

36 single benzo[a]pyrene dose group was available. Provided that a linear model is fit to continuous 

37 data, the choice of a higher BMR would not appreciably change the RPF. 
110 DRAFT – DO NOT CITE OR QUOTE 

1 Selection of point of departure. The point of departure selected for slope estimation was 

2 the BMD estimate rather than the lower confidence limit on the benchmark dose (BMDL). The 

3 BMD, as the central or “best” estimate of the dose associated with the selected BMR, was 

4 considered a more stable basis for comparison between the potency of the selected PAH and 

5 benzo[a]pyrene, and thus for calculation of relative potency, than the lower confidence limit. 

6 Extrapolation from point of departure. The slopes of the dose-response curves in the 

7 low-dose regions were calculated by linear extrapolation to the origin from the model-predicted 

8 points of departure. Equation 5-2 below shows the calculation of slope from multidose quantal 

9 data. 

10 

11 Slope = [0.1/ED10] (5-2) 

12 

13 Equation 5-3 below shows the calculation of slope from multidose continuous data. 

14 

15 Slope = [1SDchange]/[ED1SD] (5-3) 

16 

17 5.4. RPF CALCULATION FOR SINGLE DOSE DATASETS 

18 A number of studies reported data for only single doses of benzo[a]pyrene and other 

19 PAHs; for these studies, a point estimate approach was used to calculate the RPF. A point 

20 estimate approach was also used to calculate RPFs for multidose datasets when model fit was not 

21 achieved, when variance data were not available for continuous data, or when problems with 

22 model implementation were encountered. 

23 Selection of point of departure. When only one dose of each compound was used, there 

24 was only one choice for the point of departure. However, when multidose data were available, 

25 but a point estimate approach was used, the point of departure was chosen as follows. For tumor 

26 bioassay data, the lowest dose associated with a statistically significant increase in tumor 

27 incidence or multiplicity over control values was selected as the point of departure. Variance 

28 was not reported for tumor multiplicity data in any of the dermal studies and for some of the 

29 intraperitoneal studies, so the corresponding incidence data were used to determine the dose at 

30 which a significant difference from control was observed. 

31 The benzo[a]pyrene dose chosen in most instances was the lowest dose associated with a 

32 significant increase in tumor count or incidence. For tumor multiplicity data, the PAH dose 

33 chosen for the point estimate RPF calculation was the lowest dose associated with a tumor count 

34 similar to that observed at the selected benzo[a]pyrene dose (similar to selecting a BMR similar 

35 to the benzo[a]pyrene incidence). In the case of two dermal initiation studies conducted by 

36 Cavalieri et al. (1991), however, the tumor count at the lowest dose of DBalP was much higher 

37 than the tumor count at the lowest benzo[a]pyrene dose associated with statistical significance. 

38 In order to compare the doses associated with similar tumor counts (i.e., at a similar place on the 
111 DRAFT – DO NOT CITE OR QUOTE 

1 dose-response curve), a higher benzo[a]pyrene dose was chosen for the RPF calculation. A 

2 comparison of the RPFs calculated using this approach with RPFs calculated using the lowest 

3 dose associated with a statistically significant increase over controls for both dibenzo[a,l]pyrene 

4 and benzo[a]pyrene showed only small differences in the RPF values (9 vs. 10 in the 16-week 

5 study and 39 vs. 42 in the 27-week study). A similar approach was used to calculate the RPF for 

6 BjAC using the intraperitoneal multiplicity data from Mass et al. (1993). 

7 For cancer-related endpoint data, statistical analysis was not always available for each 

8 dose group. For these data, the lowest dose that produced a near maximal change in the assay of 

9 concern was selected as the point of departure. That is, the highest dose in the linear portion of 

10 the dose-response curve (identified by visual display of the data) was selected in these cases. 

11 Extrapolation from point of departure. As with multiple dose slope estimations, point 

12 estimate slope calculations also used the extra risk form. Thus, for single dose quantal data, the 

13 slope was calculated by linear extrapolation to the origin after an extra risk adjustment of the 

14 observed response (eq 5-4): 

15 

16 Slope = [(Response at dose -Control Response)÷(1-Control Response)]/Dose (5-4) 

17 

18 For single dose continuous data, the slope was calculated by linear extrapolation to the 

19 origin after adjustment of the observed response in the PAH-treated animals for the control 

20 response (eq 5-5). 

21 

22 Slope = [(Value of variable at dose) - (Value of variable)control]/Dose (5-5) 

23 

24 5.5. DOSE CONVERSION FOR RPF CALCULATION 

25 Some of the studies used to calculate RPFs reported doses or test concentrations on a 

26 molar basis (e.g., μmol per mouse, μmol/L), rather than a mass basis (mg or μg). The molar 

27 ratio differs from the mass ratio for any PAH with a molecular weight that differs from that of 

28 benzo[a]pyrene; thus, for these compounds, an RPF expressed on a mass basis will differ from 

29 that expressed on a molar basis. Table 5-1 below shows a hypothetical example for 

30 fluoranthene, a PAH with a molecular weight that differs from benzo[a]pyrene by 20%. As the 

31 table shows, the RPF differs depending on which dose units are used. 

32 

Table 5-1 Comparison between molar and mass-based RPF 

Response Dose in mol 

Molecular 

weight 

(g/mol) Dose in g Molar RPF Mass RPF 

FA 0.1 5 202.26 1,011 0.20 0.25 

BaP 0.1 1 252.32 252 1 1 

33 
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1 In order to ensure that comparisons across endpoints used consistent units, the doses used 

2 to calculate RPFs were converted to mass-based units using the molecular weight of the relevant 

3 PAH prior to estimating the RPF. The mass-based RPF was selected to be consistent with dose 

4 metrics used to calculate cancer risk; RPFs are used with oral slope factors and inhalation unit 

risks reported on a mass basis (e.g., [mg/kg-day]-1; [μg/m3]

-1 5 ). 

6 

7 5.6. SPECIAL CONSIDERATIONS FOR RPF CALCULATION USING TUMOR 

8 BIOASSAY DATA 

9 Several dermal bioassays reported significant mortality prior to the appearance of the first 

10 skin tumor. For these data sets, an assumption was made that the number of animals at risk for 

11 tumor development was equal to the total number of animals alive at the time of the appearance 

12 of the first tumor. Benign and malignant tumor types within the same target organ were 

13 combined for calculation of the RPF. The total incidence of animals with either a benign or 

14 malignant lesion was directly reported in each study (i.e., the number of animals with adenoma 

15 or carcinoma). 

16 Tumor incidence data reported for different target organs within the same group of 

17 animals were analyzed separately unless the joint incidence (incidence of either tumor type in 

18 each dose group) was reported in the publication. Liver and lung tumors were reported in 

19 newborn mice exposed to PAHs by intraperitoneal injection (LaVoie et al., 1994, 1987; Busby et 

20 al., 1989, 1984; Weyand and LaVoie, 1988; Wislocki et al., 1986). In most studies, tumor 

21 incidence was reported separately for the different target organs and could not be combined as 

22 the joint incidence was unknown. A gender difference was observed in the newborn mouse 

23 studies, with liver tumors observed in male mice only, and lung tumors reported for both male 

24 and female mice. The tumor incidence data were, therefore, evaluated separately for male and 

25 female mice. RPF values were calculated separately for male and female mice and for lung 

26 tumor incidence and liver tumor incidence in these studies. 

27 

28 5.7. SPECIAL CONSIDERATIONS FOR RPF CALCULATION USING CANCER29 RELATED ENDPOINT DATA 

30 The in vitro studies of cancer-related endpoints included measurements of bacterial 

31 mutagenicity, mammalian mutagenicity, morphological/malignant cell transformation, DNA 

32 adduct formation, DNA damage or repair, and clastogenicity or sister chromatid exchange 

33 frequency. Many of the studies describing in vitro cancer-related endpoints provide dose34 response data under varying study conditions. For example, bacterial mutagenesis studies used 

35 multiple strains, different metabolic activation processes, and/or varying assay systems. In order 

36 to limit the number of datasets used for dose-response analysis of in vitro mutagenicity studies, 

37 and to provide a consistent basis for comparing RPFs for differenct PAHs, data associated with 

38 the conditions that maximized the benzo[a]pyrene response within a particular study were used 
113 DRAFT – DO NOT CITE OR QUOTE 

1 for the dose-response assessment of PAHs. It should be noted that in several studies, test 

2 conditions that were optimal for benzo[a]pyrene were not necessarily optimal for the selected 

3 PAH (see Appendix C for specific studies). The uncertainties and limitations associated with 

4 this approach are discussed further in Section 8. 

5 For time-course studies of DNA adducts, results were reported as either AUC or peak 

6 formation of adducts. AUC was considered preferable for dose-response assessment, because 

7 this measure considers both adduct formation and repair. Adducts measured in more than one 

8 organ were summed to derive a total measure of adduct formation (standardized per unit amount 

9 of DNA). 

10 The data for bacterial and mammalian cell mutagenicity and malignant cell 

11 transformation were sometimes expressed as a mutation or transformation frequency (i.e., 

12 mutants/total cell count or transformed cells/total cells). For multiple-dose studies, these quantal 

13 variables were evaluated using the multistage model as described above. Problems were 

14 sometimes encountered when using the multistage model for incidence data of this type. In some 

15 cases, modifying the initial parameters in the multistage algorithm facilitated convergence. In a 

16 select few cases, the quantal linear model was used when the multistage model would not 

17 converge. If neither the multistage nor quantal linear models provided adequate fit, a point 

18 estimate approach was used. If possible, the point estimates for both benzo[a]pyrene and the 

19 target PAH were chosen at a comparable response level (e.g., the doses of benzo[a]pyrene and 

the target PAH that both gave two mutants in 105 20 cells). However, in many cases, a comparable 

21 response rate was not available. In these instances, the RPF was derived from slopes calculated 

22 by linear extrapolation from the peak response. 

23 As noted earlier, for studies that included only one dose of benzo[a]pyrene and multiple 

24 dose data for a selected PAH, the BMR selected for dose-response modeling for the selected 

25 PAH was the benzo[a]pyrene response with the background or control response subtracted. In 

26 some instances, when the benzo[a]pyrene response level greatly exceeded the response at the 

27 highest dose of the selected PAH, the software would fail to calculate the ED at the 

28 benzo[a]pyrene response level. In these instances, a point estimate approach using the peak 

29 response for the selected PAH was used. 

30 The individual study RPFs calculated for each PAH were used in a weight of evidence 

31 evaluation to assess the potential carcinogenicity of each compound (see Section 6) and in the 

32 derivation of a final RPF for each compound (Section 7). 

33 

34 

35 6. SELECTION OF PAHS FOR INCLUSION IN RELATIVE POTENCY APPROACH 

36 

37 
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1 The selection of PAHs to be included in the RPF approach began with an evaluation of 

2 whether the available data were adequate to assess the potential carcinogenicity of each 

3 compound. At least one RPF value was calculated for each of 50 PAHs. For 16 of these 

4 compounds, only a single RPF value derived from an in vitro cancer-related endpoint (primarily 

5 mutagenicity assays) was available. These PAHs are shown in Table 6-1. Due to the limited 

6 data available for these 16 compounds, no further evaluation of these PAHs was conducted, and 

7 they were not selected for inclusion in the RPF approach. 

8 

Table 6-1. PAHs with only one RPF from in vitro cancer-related endpoint 

study and excluded from RPF approach 

PAH CASRN Abbreviation 

Aceanthrylene 202-03-09 ACEA 

Acenaphthene 83-32-9 AN 

Acenaphthylene 208-96-8 ANL 

Acephenanthrylene 201-06-9 APA 

Benzo[a]perylene 191-85-5 BaPery 

Benz[b]anthracene 92-24-9 BbA 

Benzo[b]perylene 197-70-6 BbPery 

Benzo[c]phenanthrene 195-19-7 BcPH 

Cyclopent[h,i]aceanthrylene 131581-33-4 CPhiACEA 

Cyclopent[h,i]acephenanthrylene 114959-37-4 CPhiAPA

Dibenzo[a,f]fluoranthene 203-11-2 DBafF 

Dibenz[a,j]anthracene 224-41-9 DBajA 

Dibenzo[b,e]fluoranthene 2997-45-7 DBbeF 

Dibenzo[e,l]pyrene 192-51-8 DBelP 

Dibenz[k,mno]acephenanthrylene 153043-81-3 DBkmnoAPH 

Naphtho[2,3-a]pyrene 196-42-9 N23aP 

9 

10 The remaining 34 PAHs had RPF values calculated from at least one in vivo dataset or at 

11 least two in vitro cancer-related endpoint datasets. For these compounds, a weight of evidence 

12 approach was used to determine whether the available data (including the calculated RPFs as 

13 well as negative studies that met selection criteria) were adequate to assess the carcinogenic 

14 potential. Using the calculated RPFs in the weight of evidence evaluation allowed consideration 

15 of the magnitude of calculated RPFs in assessing potential carcinogenicity. When data were not 

16 considered adequate, the PAH was excluded from the RPF approach. When data were 

17 considered adequate for a given PAH, it was selected for inclusion. 

18 A PAH with adequate evidence to suggest that it has little or no carcinogenic potential 

19 was selected for inclusion in the RPF approach and assigned an RPF of 0. While there is little 

20 quantitative difference between selecting a final RPF of zero for a given PAH and excluding that 

21 PAH from the RPF approach, this is an important distinction for uncertainty analysis. There is 
115 DRAFT – DO NOT CITE OR QUOTE 

1 substantial uncertainty in the risk associated with a PAH that is excluded from the RPF approach 

2 due to inadequate data; this compound could be of low or high potency. However, for a PAH 

3 with an RPF of 0, there is evidence to suggest that this compound exhibits little or no 

4 carcinogenic potential, and the uncertainty associated with the cancer risk for this compounds is 

5 markedly reduced. For anthracene, phenanthrene, and pyrene, it has been determined that the 

6 available data support a practical RPF of zero. It is possible that the studies available may not 

7 provide sufficient sensitivity to compare the potency of the PAHs of interest to benzo[a]pyrene, 

8 and thus, the RPF of zero should not be considered a characterization of the inherent 

9 carcinogenicity of anthracene, phenanthrene, or pyrene. The weight of evidence analysis is 

10 outlined in Section 6.1 and the results are described in narratives for each of the 34 individual 

11 PAHs (Section 6.2). Section 7 describes how the RPFs from multiple datasets were used to 

12 derive final RPFs for those PAHs selected for inclusion in the approach, and reports the final 

13 RPF information for each PAH. 

14 

15 6.1. METHOD FOR SELECTING PAHS FOR INCLUSION IN RELATIVE POTENCY 

16 APPROACH 

17 For each of the 34 PAHs, a weight of evidence evaluation was conducted to assess the 

18 evidence that each PAH could induce a carcinogenic response. This evaluation did not constitute 

19 a formal weight of evidence evaluation of carcinogenic potential; rather, an approach was 

20 developed using the data collected for this analysis to determine whether the available 

21 information for each PAH was adequate to draw a conclusion regarding carcinogenic potential. 

22 When the data were considered adequate for a given PAH, it was selected for inclusion in the 

23 RPF approach. Figure 6-1 shows the decision tree that was used to evaluate the data for each 

24 PAH and to determine whether it should be included in the RPF approach. The weight of 

25 evidence evaluation concluded with one of two possible outcomes: 

26 

27 1. The data reviewed are adequate to evaluate potential carcinogenicity and the PAH should 

28 be included in the RPF analysis, or 

29 

30 2. The data reviewed are inadequate to assess carcinogenic potential and the PAH should be 

31 excluded from the RPF analysis. 

32 
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Has PAH been tested in 

tumor bioassay with 

BaPa?

Did PAH give positive 

result in any tumor 

bioassays with BaP?

Did PAH give 

positive result 

in all tumor 

bioassays with 

BaP?

Can conflicting 

tumor bioassay 

results be 

explained by 

differences in 

study design?

Do other tumor bioassaysb

and/or cancer-related 

endpoint datac provide 

adequate data to assess 

carcinogenic potential?

Data are 

adequate to 

assess 

carcinogenic 

potential of 

PAH: 

Include in RPF 

approach

Data are 

inadequate to 

assess carcinogenic 

potential of PAH: 

Exclude from 

RPF approach

No

No

No

No

No

Yes

Yes

Yes

Yes

Yes

1 

2 

a

3 Bioassays with benzo[a]pyrene that met study quality criteria (includes studies with 

4 negative results). 

b

5 Other bioassays include those that did not test benzo[a]pyrene and/or those that were not 

6 suitable for RPF derivation (e.g., incidence at lowest dose exceeded 90%). 

c

7 Cancer-related endpoint data examined in this process included studies of DNA adducts, 

8 clastogenicity or sister chromatid exchange, mutagenicity, morphological transformation, 

9 DNA damage, unscheduled DNA synthesis, etc. that included the selected PAH and 

10 benzo[a]pyrene. 

11 

12 Figure 6-1. Weight of evidence analysis of potential carcinogenicity. 

13 

14 In vivo tumor bioassays that included benzo[a]pyrene were given the greatest weight in 

15 assessing the potential carcinogenicity of a given PAH; data from other bioassays and cancer-
117 DRAFT – DO NOT CITE OR QUOTE 

1 related endpoint studies were used to supplement the weight of evidence when the bioassay data 

2 that included benzo[a]pyrene were conflicting or negative. Structural alerts for PAH 

3 carcinogenicity or mutagenicity (specifically, at least four aromatic rings, or the presence of a 

4 classic bay region or fjord region formed entirely by aromatic rings) were noted in the evaluation 

5 for each PAH, but were not used explicitly in the weight of evidence evaluation. 

6 When there were bioassays including benzo[a]pyrene with positive findings, and none 

7 with negative findings for a given PAH, that compound was selected for inclusion in the RPF 

8 approach, and no further evaluation of cancer-related endpoint data was conducted. However, 

9 the cancer-related endpoint findings for these compounds were noted in the individual PAH 

10 narratives (Section 6.2). Among the PAHs included in this analysis, there were none with 

11 positive bioassay data and robust negative cancer-related endpoint data. Were this instance to 

12 arise, it would require special consideration, as it might imply a different mode of carcinogenic 

13 action than the PAHs addressed herein. 

14 Bioassays that met selection criteria (see Section 4.3) were included in the weight of 

15 evidence analysis, regardless of whether positive or nonpositive (i.e., negative) results were 

16 found. However, the weight of evidence evaluation assumed that a given compound may be 

17 active in one system (e.g., newborn mouse), and inactive or weakly active in another (e.g., 

18 dermal initiation). Thus, when conflicting results were observed in different test systems, 

19 different species, or different genders, the PAH was assumed to be potentially carcinogenic 

20 based on the positive findings and was included in the RPF approach. 

21 In order to compare the results of bioassays with positive and nonpositive results in the 

22 same test system, an “RPF detection limit” was conceptualized as a means of approximating the 

23 minimum RPF that could be determined under the conditions of the study. The “RPF detection 

limit” was defined as the ratio of the dose-response slopes324 , using the lowest statistically 

25 significant response that could be calculated for the subject PAH and the actual benzo[a]pyrene 

26 response, as points of departure for the slope calculation. The lowest statistically significant 

27 response was calculated using the incidence of tumors in the control group, number of animals in 

the group treated with the subject PAH, and Fisher’s exact test428 (employing a one-sided 

29 p-value ≤ 0.05). Appendix F shows an example calculation of an “RPF detection limit.” The 

30 utility of this concept is in weighing positive and nonpositive bioassay results. If all of the 

31 nonpositive studies had “RPF detection limits” in excess of what is observed in the positive 

32 studies, then it is plausible that the nonpositive studies may not have been sufficiently sensitive 

33 to estimate the low RPF appropriate to the subject PAH. In this event, the PAH was considered 

34 potentially carcinogenic and included in the RPF approach. 

 

3

The standard RPF equation is RPF = slope PAHi ÷ slope BaP = [response/dose]PAHi ÷ [response/dose]BaP. 

4

This calculation was implemented using trial and error within the Fisher’s exact test in the online statistical 

calculator GraphPad©. 
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1 If there were no bioassays with benzo[a]pyrene for a given compound, all of the selected 

2 bioassays gave nonpositive results, or inconsistent results could not be explained by test system 

3 or “RPF detection limit”, then the results of other bioassays (those without benzo[a]pyrene, or 

4 those rejected from dose-response assessment exclusively because of concerns associated with 

5 benzo[a]pyrene) and cancer-related endpoint data were evaluated. The weight of evidence 

6 analysis then considered all of the following information: bioassays with benzo[a]pyrene, other 

7 bioassays, and cancer-related endpoint data. If these data were determined to be inadequate to 

8 assess the carcinogenic potential for a given PAH, then that compound was excluded from the 

9 RPF approach. If the data were considered adequate to assess the carcinogenic potential, the 

10 compound was retained and a final RPF was derived. Section 6.2 below describes the weight of 

11 evidence evaluation for each of the 34 PAHs. Section 7.1 describes how final RPFs were 

12 derived for the 26 PAHs selected for inclusion in the RPF approach. 

13 

14 6.2. WEIGHT OF EVIDENCE EVALUATION FOR 34 INDIVIDUAL PAHS 

15 For each PAH, the structure is shown along with a brief reference to any structural alerts 

16 for potential carcinogenicity (specifically, more than three aromatic rings and/or bay or fjord 

17 region in alternant PAH). Next, a brief narrative describing the weight of evidence evaluation is 

18 given, with a graphical representation of the data that were available for RPF calculation 

19 (Figures 6-2 to 6-35). The graph for each compound provides a visual representation of the 

20 database of studies that included both the subject PAH and benzo[a]pyrene. The solid bars show 

21 the values of the RPFs calculated from all studies with positive findings. The x-axis label shows 

22 the reference for the pertinent study. The RPFs are color-coded to distinguish among in vivo 

23 tumor bioassays based on incidence data, in vivo tumor bioassays based on multiplicity data, in 

24 vivo cancer-related endpoint studies, and in vitro cancer-related endpoint studies. Within these 

25 categories, the RPFs are ordered (left to right in the graph) from highest to lowest, with positive 

26 results shown before nonpositive results. 

27 For each nonpositive bioassay, an empty, dotted bar shows what is termed the “RPF 

28 detection limit” (see Section 6.1 for description). Missing bars designate cancer-related studies 

29 that resulted in nonpositive findings. An RPF detection limit for nonpositive cancer-related 

30 studies was not included, because comparisons between nonpositive and positive studies were 

31 complicated by the wide variety of study conditions (e.g., test species and strains, metabolic 

32 activation sources, assay systems). 

33 Each narrative concludes with a statement as to whether the subject PAH was selected for 

34 inclusion in the PAH RPF approach. The weight of evidence evaluation for the 34 PAHs with at 

35 least one in vivo RPF or at least 2 in vitro cancer-related endpoint RPFs resulted in the selection 

36 of 26 PAHs for inclusion in the RPF approach and the exclusion of 8 PAHs from the approach 

37 (see Table 6-2). 

38 
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1 

Table 6-2. Results of weight of evidence evaluation of 34 PAHs 

Adequate data: selected for inclusion in RPF approach 

PAH CASRN Abbreviation PAH CASRN Abbreviation 

Benzo[a]pyrene 50-32-8 BaP Cyclopenta[d,e,f]chrysene, 

4H- 202-98-2 CPdefC 

Anthanthrene 191-26-4 AA Dibenz[a,c]anthracene 215-58-7 DBacA 

Anthracene 120-12-7 AC Dibenzo[a,e]fluoranthene 5385-75-1 DBaeF 

Benz[a]anthracene 56-55-3 BaA Dibenzo[a,e]pyrene 192-65-4 DBaeP 

Benz[b,c]aceanthrylene, 

11H- 202-94-8 BbcAC Dibenz[a,h]anthracene 53-70-3 DBahA 

Benzo[b]fluoranthene 205-99-2 BbF Dibenzo[a,h]pyrene 189-64-0 DBahP 

Benz[e]aceanthrylene 199-54-2 BeAC Dibenzo[a,i]pyrene 189-55-9 DBaiP 

Benzo[g,h,i]perylene 191-24-2 BghiP Dibenzo[a,l]pyrene 191-30-0 DBalP 

Benz[j]aceanthrylene 202-33-5 BjAC Fluoranthene 206-44-0 FA 

Benzo[j]fluoranthene 205-82-3 BjF Indeno[1,2,3-c,d]pyrene 193-39-5 IP 

Benzo[k]fluoranthene 207-08-9 BkF Naphtho[2,3-e]pyrene 193-09-9 N23eP 

Benz[l]aceanthrylene 211-91-6 BlAC Phenanthrene 85-01-8 PH 

Chrysene 218-01-9 CH Pyrene 129-00-0 Pyr 

Cyclopenta[c,d]pyrene 27208-37-3 CPcdP 

Inadequate data 

PAH CASRN Abbreviation PAH CASRN Abbreviation 

Acepyrene, 2,3- 25732-74-5 ACEP Coronene 191-07-1 CO 

Benzo[b]fluorene, 11H- 243-17-4 BbFE Fluorene 86-73-7 FE 

Benzo[e]pyrene 192-97-2 BeP Perylene 198-55-0 Pery 

Benzo[g,h,i]fluoranthene 203-12-3 BghiF Triphenylene 217-59-4 Tphen 

2 
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1 

2 2,3-Acepyrene (ACEP) 

3 

4 

5 2,3-Acepyrene (CASRN 25732-74-5) is a nonalternant PAH comprised of four aromatic 

6 rings and one five-membered ring. 2,3-Acepyrene does not contain a classic bay or fjord region 

7 in its structure. 

8 Five datasets for 2,3-acepyrene met selection criteria and included benzo[a]pyrene 

9 (shown in Figure 6-2). Dermal initiation and complete carcinogenicity bioassays in mice both 

10 resulted in nonpositive findings (both published by Cavalieri et al., 1981b). RPF detection limits 

11 for these studies were 0.09 and 0.02, respectively. The limited cancer-related data are mixed, 

12 with one positive dataset for in vivo DNA adduct formation, one positive bacterial mutagenicity 

13 dataset (both published by Cavalieri et al., 1981a), and one negative mammalian mutagenicity 

14 dataset (Barfknecht et al., 1982). There are no bioassays of 2,3-acepyrene without 

15 benzo[a]pyrene. Overall, the database for 2,3-acepyrene is both limited and inconsistent. The 

16 database for 2,3-acepyrene does not provide adequate information with which to assess potential 

17 carcinogenicity; this PAH was not selected for inclusion in the RPF approach. 

18 
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1 2 Figure 6-2. 2,3-Acepyrene (ACEP) RPFs*. 

0.001

0.01

0.1

1

10

100

Cavalieri et al., 1981b Cavalieri et al., 1981b Cavalieri et al., 1981a Cavalieri et al., 1981a Barfknecht et al., 198

Value of RPF

Reference

RPF detection limits 

for nonpositive 

bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Anthanthrene (AA) 

2 

3 

4 

5 Anthanthrene (CASRN 191-26-4) is an alternant PAH comprised of six fused aromatic 

6 rings. Anthanthrene does not have a bay or fjord region in its structure. 

7 There are seven datasets for anthanthrene that met selection criteria and included 

8 benzo[a]pyrene (Figure 6-3). The database includes three in vivo tumor bioassays, three 

9 bacterial mutagenicity datasets, and one in vitro DNA damage dataset. Statistically increased 

10 tumor incidences were reported in both a rat lung implantation bioassay (Deutsch-Wenzel et al., 

11 1983) and a dermal complete carcinogenicity bioassay in mice (Cavalieri et al., 1977). No 

12 increase over control tumor incidence was reported in a dermal initiation study (Hoffmann and 

13 Wynder, 1966), but the RPF detection limit for this study was 0.3. All of the cancer-related 

14 endpoint studies gave positive results. Because conflicting bioassay data can be explained by 

15 differences in study design (initiation versus complete dermal carcinogenicity), anthanthrene was 

16 considered potentially carcinogenic and selected for inclusion in the RPF approach. 

17 
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1 2 Figure 6-3. Anthanthrene (AA) RPFs. 

0.001

0.01

0.1

1

10

100

Cavalieri et al., 1977 Deutsch-Wenzel et al., 

1983

Hoffman and Wynder, 

1966

Andrews et al., 1978 Hermann, 1981 Mersch-Sundermann et 

al., 1992

Kaden et al., 

Value of RPF

Reference

RPF detection limit 

for nonpositive 

bioassay

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
124 DRAFT – DO NOT CITE OR QUOTE 

1 Anthracene (AC) 

2 

3 

4 

5 Anthracene (CASRN 120-12-7) is an alternant PAH comprised of three fused aromatic 

6 rings. Anthracene does not have a bay or fjord region in its structure, and contains less than four 

7 aromatic rings. 

8 Thirty-seven datasets for anthracene met selection criteria and included benzo[a]pyrene, 

9 including 1 dermal initiation tumor bioassay, 3 in vivo clastogenicity or sister chromatid 

10 exchange datasets, 10 bacterial mutagenicity datasets, 4 mammalian mutagenicity datasets, 

11 6 morphological/malignant cell transformation datasets, and 13 in vitro DNA adduct, DNA 

12 damage or clastogenicity datasets (Figure 6-4). The single dermal initiation bioassay gave a 

13 nonpositive result, with a RPF detection limit of 0.2 (LaVoie et al., 1985). Only two datasets 

14 gave positive results: an in vitro bacterial mutagenicity assay and an in vitro study of DNA 

15 damage. The remaining 35 datasets reported nonpositive findings. To confirm the negative 

16 findings in the one tumor bioassay that included benzo[a]pyrene, other bioassays and cancer17 related endpoint data for anthracene were considered in the weight of evidence evaluation. In 

18 bioassays without benzo[a]pyrene, anthracene did not induce a statistically significant increase in 

19 tumor incidence in two dermal initiation studies (LaVoie et al., 1983; Salaman and Roe, 1956) 

20 and a lung implantation bioassay (Stanton, 1972). Scribner (1973) reported a weak tumorigenic 

21 response in a dermal initiation study in mice (4/28 mice developed papillomas by week 35 after 

22 dermal treatment with 10 μmol anthracene in benzene followed by twice weekly treatment with 

23 TPA, as compared with 0/30 control mice, p = 0.048). 

24 In vitro assays of mutagenicity (both bacterial and mammalian) are nearly all negative for 

25 anthracene (13/14 studies). Studies of morphological/malignant cell transformation were all 

26 negative. Finally, in numerous in vitro studies of DNA damage or clastogenicity, anthracene has 

27 given nonpositive results (12/13). Sakai et al. (1985) reported a mutagenic response in bacteria 

28 treated with anthracene, and Rossman et al. (1991) observed evidence of unscheduled DNA 

29 synthesis in E. coli treated with anthracene. Overall, the weight of evidence suggests that 

30 anthracene is not carcinogenic or is of very low carcinogenic potential. In addition, anthracene 

31 lacks all three known structural alerts (at least four rings, bay region or fjord region) for PAH 

32 carcinogenicity and/or mutagenicity. Because the weight of evidence evaluation suggests that 

33 the data are adequate to assess the carcinogenic potential of anthracene, this compound was 

34 selected for inclusion in the RPF approach and assigned a RPF of 0. 
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1 2 Figure 6-4. Anthracene (AC) RPFs*.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

RPF detection limits 

for nonpositive 

bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
126 DRAFT – DO NOT CITE OR QUOTE 

1 Benz[a]anthracene (BaA) 

2 

3 

4 Benz[a]anthracene (CASRN 56-55-3) is an alternant PAH comprised of four fused 

5 aromatic rings. Benz[a]anthracene contains a bay region but no fjord region in its structure. 

6 There are 65 datasets for benz[a]anthracene that met selection criteria and included 

7 benzo[a]pyrene (Figure 6-5). Included in the database are tumor bioassays (5), in vivo DNA 

8 adduct studies (4), in vivo clastogenicity studies (4), an in vivo mutagenicity study (1), bacterial 

9 mutagenicity (15), mammalian mutagenicity (14), morphological/malignant cell transformation 

10 assays (6), and in vitro studies of DNA damage, adducts, or clastogenicity (16). There are five 

11 tumor bioassay datasets of benz[a]anthracene that included benzo[a]pyrene; four gave positive 

12 results, and one gave a nonpositive result. The positive findings were in different genders tested 

13 in a newborn mouse study using intraperitoneal injection (Wislocki et al., 1986); the datasets 

14 included both tumor incidence and multiplicity data for both sexes. Positive results were also 

15 reported in a dermal initiation study (Slaga et al., 1978). The one nonpositive bioassay (Cavalieri 

16 et al., 1977) was a dermal complete carcinogenicity study with an RPF detection limit of 0.2. 

17 Benz[a]anthracene was shown to form DNA adducts when administered in vivo in both rats and 

18 mice via injection and gavage (Kligerman et al., 2002). Mutagenicity and morphological/

19 malignant cell transformation assays of benz[a]anthracene were predominantly positive, as were 

20 studies of other cancer-related endpoints. 

21 Given that the differing bioassay results can be attributed to different test systems and 

22 study design, benz[a]anthracene was considered potentially carcinogenic and was selected for 

23 inclusion in the RPF approach. 

24 
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1 2 Figure 6-5. Benz[a]anthracene (BaA) RPFs*.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

RPF detection limit 

for nonpositive 

bioassay

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicityIn vivo cancer-related endpoinIn vitro cancer-related endpoin
128 DRAFT – DO NOT CITE OR QUOTE 

1 11H-Benz[b,c]aceanthrylene (BbcAC) 

2 

3 

4 11H-Benz[b,c]aceanthrylene (CASRN 202-94-8) is a nonalternant PAH comprised of 

5 four aromatic rings and one five-membered ring. 11H-Benz[b,c]aceanthrylene does not contain a 

6 classic bay or fjord region in its structure. 

7 There was only one dataset for benz[b,c]aceanthrylene that met selection criteria and 

8 included benzo[a]pyrene (Figure 6-6). This multi-dose dermal initiation study resulted in an 

9 RPF estimate of 0.05 (Rice et al., 1988). Benz[b,c]aceanthrylene has not been tested in any 

10 bioassay without benzo[a]pyrene. There are no cancer-related endpoint data for 

11 benz[b,c]aceanthrylene. As the only available bioassay of this PAH was positive, 

12 benz[b,c]aceanthrylene was considered potentially carcinogenic and was selected for inclusion in 

13 the RPF approach. 

14 
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1 2 Figure 6-6. 11H-Benz[b,c]aceanthrylene (BbcAC) RPFs. 

0.001

0.01

0.1

1

10

100

Rice et al., 1988

Value of RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
130 DRAFT – DO NOT CITE OR QUOTE 

1 Benzo[b]fluoranthene (BbF) 

2 

3 

4 Benzo[b]fluoranthene (CASRN 205-99-2) is a nonalternant PAH comprised of four 

5 aromatic rings and one five-membered ring. Benzo[b]fluoranthene contains one classic bay 

6 region but no fjord region in its structure. 

7 There were 21 datasets of benzo[b]fluoranthene that met selection criteria and included 

8 benzo[a]pyrene (Figure 6-7). Included in the database are in vivo tumor bioassay datasets (7), in 

9 vivo DNA adduct datasets (7), in vivo clastogenicity datasets (3), mutagenicity and 

10 morphological/malignant cell transformation datasets (3), and an in vitro DNA damage dataset 

11 (1). Statistically significant increases in tumor incidence and/or multiplicity were reported in 

12 male mice tested in two newborn mouse bioassays using intraperitoneal injection (Nesnow et al., 

13 1998b; LaVoie et al., 1987), in dermal initiation (LaVoie et al., 1982) and dermal complete 

14 carcinogenicity (Habs et al., 1980) bioassays, and in a rat lung implantation bioassay (Deutsch15 Wenzel et al., 1983). The one nonpositive result was in female mice tested in the newborn 

16 mouse bioassay; the RPF detection limit was 0.8 (LaVoie et al., 1987). A number of studies 

17 showed that benzo[b]fluoranthene forms DNA adducts when administered in vivo to rats or mice 

18 via injection or gavage (Kligerman et al., 2002; Nesnow et al., 1998b, 1993b). One mutagenicity 

19 assay and two morphological/malignant cell transformation assays of benzo[b]fluoranthene were 

20 positive, as were studies of other cancer-related endpoints; there were no negative studies of 

21 cancer-related endpoints. Given that the differeing bioassay results can be attributed to different 

22 gender, benz[a]anthracene was considered potentially carcinogenic and was selected for 

23 inclusion in the RPF approach. 
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1 2 Figure 6-7. Benzo[b]fluoranthene (BbF) RPFs.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

RPF detection limit 

for nonpositive 

bioassay

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
132 DRAFT – DO NOT CITE OR QUOTE 

1 11H-Benzo[b]fluorene (BbFE) 

2 

3 11H-Benzo[b]fluorene (CASRN 243-17-4) is a nonalternant PAH comprised of three 

4 aromatic rings and one five-membered ring. 11H-Benzo[b]fluorene does not contain a classic 

5 bay or fjord region in its structure. 

6 There were three datasets for 11H-benzo[b]fluorene that met selection criteria and 

7 included benzo[a]pyrene (Figure 6-8): two mutagenicity datasets and an in vitro DNA damage 

8 dataset. There are no bioassays of 11H-benzo[b]fluorene that included benzo[a]pyrene, so 

9 bioassays without benzo[a]pyrene and cancer-related endpoint data were considered. LaVoie et 

10 al. (1981) conducted a study of skin tumor initiation in mice treated with 1.0 mg 11H-benzo[b]

11 fluorene followed by 20 weeks of treatment with TPA. The incidence of tumor-bearing animals 

12 (4/20) was not significantly increased over controls (1/20) (LaVoie et al., 1981). The limited 

13 cancer-related endpoint data were mixed, with one positive mutagenicity study (Kaden et al., 

14 1979), one negative mutagenicity study (Hermann, 1981), and one positive in vitro study of 

15 DNA damage (Mersch-Sundermann et al., 1992). Overall, the database for 11H-benzo[b]

16 fluorene is both limited and inconsistent. Because the database for 11H-benzo[b]fluorene does 

17 not provide adequate information with which to assess potential carcinogenicity, this PAH was 

18 not selected for inclusion in the RPF approach. 

19 
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1 2 Figure 6-8. 11H-Benzo[b]fluorene (BbFE) RPFs*.

0.001

0.01

0.1

1

10

100

Kaden et al., 1979 Mersch-Sundermann et al., 1992 Hermann, 1981

Value of RPF

Reference * Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
134 DRAFT – DO NOT CITE OR QUOTE 

1 Benz[e]aceanthrylene (BeAC. 

2 

3 Benz[e]aceanthrylene (CASRN 199-54-2) is a nonalternant PAH comprised of four 

4 aromatic rings and one five-membered ring. Benz[e]aceanthrylene contains a classic bay region 

5 but no fjord region in its structure. 

6 There were six datasets for benz[e]aceanthrylene that met selection criteria and included 

7 benzo[a]pyrene (Figure 6-9); all gave positive results. The database includes an in vivo tumor 

8 bioassay in two sexes (each reporting both incidence and multiplicity), a mammalian 

9 mutagenicity study, and a morphological/malignant cell transformation study. Significantly 

10 increased tumor incidence and tumor multiplicity were reported for both male and female mice 

11 in a dermal initiation bioassay in mice (Nesnow et al., 1984). As the available bioassay that 

12 included benzo[a]pyrene was positive, benz[e]aceanthrylene was considered potentially 

13 carcinogenic and was selected for inclusion in the RPF approach. 

14 
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1 2 Figure 6-9. Benz[e]aceanthrylene (BeAC) RPFs.

0.001

0.01

0.1

1

10

100

Nesnow et al., 1984 Nesnow et al., 1984 Nesnow et al., 1984 Nesnow et al., 1984 Nesnow et al., 1984b Mohapatra et al., 1987

Value of RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
136 DRAFT – DO NOT CITE OR QUOTE 

1 Benzo[e]pyrene (BeP) 

2 

3 

4 Benzo[e]pyrene (192-97-2) is an alternant PAH comprised of five fused aromatic rings. 

5 Benzo[e]pyrene contains two bay regions and no fjord region in its structure. 

6 Thirty-seven datasets for benzo[e]pyrene met selection criteria and included 

7 benzo[a]pyrene: 2 tumor bioassays, 1 in vivo clastogenicity dataset, 12 bacterial mutagenicity 

8 datasets, 4 mammalian mutagenicity datasets, 7 morphological/malignant cell transformation 

9 datasets, and 11 in vitro DNA damage or clastogenicity datasets (Figure 6-10). No increase in 

10 tumor incidence was observed when benzo[e]pyrene was tested alone as part of a dermal 

11 cocarcinogenicity bioassay (Van Duuren and Goldschmidt, 1976). When tested in a lung 

12 implantation bioassay in rats, benzo[e]pyrene exposure did not result in a significant increase in 

13 tumor incidence (Deutsch-Wenzel et al., 1983). The RPF detection limits of these studies were 

14 approximately 0.01 and 0.1. To confirm the negative findings in the available tumor bioassays 

15 that included benzo[a]pyrene, other bioassays and cancer-related endpoint data were considered. 

16 In bioassays without benzo[a]pyrene, benzo[e]pyrene gave negative results in a dermal initiation 

17 bioassay (1 mg/mouse; Van Duuren et al., 1968) and a newborn mouse bioassay (0.7 μmol; 

18 Chang et al., 1981). A significant increase in tumor incidence was reported in a single19 concentration dermal initiation study in mice; 11/13 surviving mice (20 were treated) had 

20 papillomas by week 35 after dermal treatment with 10 μmol benzo[e]pyrene in benzene 

21 (p < 0.0001), followed by twice weekly treatment with TPA; no control mice had papillomas 

22 (Scribner, 1973). 

23 In vitro assays of mutagenicity (both bacterial and mammalian) and morphological/

24 malignant cell transformation give inconsistent results for benzo[e]pyrene; 11/23 studies were 

25 positive and the rest were negative. Positive studies include a mix of bacterial mutagenicity and 

26 morphological/malignant cell transformation assays; four mammalian mutagenicity assays were 

27 negative. One study of in vivo clastogenicity and two studies of in vitro DNA damage were 

28 positive, while nine studies of in vitro DNA damage or clastogenicity were negative. 

29 While the database for benzo[e]pyrene is quite large, the results are inconsistent; as a 

30 result, no conclusion can be drawn as to potential carcinogenicity. This PAH was not selected 

31 for inclusion in the RPF approach. 
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1 2 Figure 6-10. Benzo[e]pyrene (BeP) RPFs*.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

RPF detection 

limits for 

nonpositive 

bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
138 DRAFT – DO NOT CITE OR QUOTE 

1 Benzo[g,h,i]fluoranthene (BghiF) 

2 

3 

4 Benzo[g,h,i]fluoranthene (CASRN 203-12-3) is a nonalternant PAH comprised of four 

5 aromatic rings and one five-membered ring. Benzo[g,h,i]fluoranthene does not contain a classic 

6 bay or fjord region in its structure. 

7 There were six datasets for benzo[g,h,i]fluoranthene that met selection criteria and 

8 included benzo[a]pyrene (Figure 6-11). A dermal initiation bioassay in mice (Van Duuren et al., 

9 1966) did not result in a statistically significant increase in tumor incidence; the RPF detection 

10 limit was 0.06. There were no other bioassays that met selection criteria. There were three 

11 positive bacterial mutagenicity studies (Chang et al., 2002; Lafleur et al., 1993; Carver et al., 

12 1986), one positive study of in vitro DNA damage (Mersch-Sundermann et al., 1992), and a 

13 mammalian mutagenicity study with negative results (Lafleur et al., 1993). The RPF values for 

14 the positive cancer-related endpoint datasets ranged from 0.6 to 1. Overall, the database for 

15 benzo[g,h,i]fluroanthene is both limited and inconsistent. Because the database for 

16 benzo[g,h,i]fluoranthene does not provide adequate information with which to assess potential 

17 carcinogenicity, this PAH was not selected for inclusion in the RPF approach. 

18 
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1 2 Figure 6-11. Benzo[g,h,i]fluoranthene (BghiF) RPFs*.

0.001

0.01

0.1

1

10

100

Van Duuren et al., 1966 LaFleur et al., 1993 Carver et al., 1986 Chang et al., 2002 Mersch-Sundermann et al., 

1992

LaFleur et al., 1993

Value of RPF

Reference

RPF detection limits 

for nonpositive 

bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
140 DRAFT – DO NOT CITE OR QUOTE 

1 Benzo[g,h,i]perylene (BghiP) 

2 

3 

4 Benzo[g,h,i]perylene (CASRN 191-24-2) is an alternant PAH comprised of six fused 

5 aromatic rings. Benzo[g,h,i]perylene contains a bay region but no fjord region in its structure. 

6 There were 10 datasets for benzo[g,h,i]perylene that met selection criteria and included 

7 benzo[a]pyrene (Figure 6-12). The database includes three in vivo tumor bioassays, four 

8 bacterial mutagenicity datasets, an in vitro DNA damage dataset, and two in vitro DNA adduct 

9 datasets. Of the three bioassays, positive findings were only reported in one: a rat lung 

10 implantation bioassay (Deutsch-Wenzel et al., 1983) that resulted in a RPF estimate of 0.009. In 

11 a dermal initiation bioassay (Hoffmann and Wynder, 1966) and a dermal cocarcinogenicity 

12 bioassay (Van Duuren and Goldschmidt, 1976), there was no statistically significant increase in 

13 tumor incidence, but these studies had relatively insensitive RPF detection limits (around 0.1) 

14 compared with the positive study. There were four positive mutagenicity studies; all were 

15 conducted in bacterial systems. Studies of in vitro DNA adducts and DNA damage were 

16 positive. Because the inconsistent bioassay results can be attributed to different test systems 

17 (different species and route), benzo[g,h,i]perylene was considered potentially carcinogenic and 

18 was selected for inclusion in the RPF approach. 
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1 2 Figure 6-12. Benzo[g,h,i]perylene (BghiP) RPFs*.

0.001

0.01

0.1

1

10

100

Deutsch-Wenzel 

et al., 1983

Hoffmann and Wynder, 1966

Van Duuren and 

Goldschmidt, 

1976

Andrews et al., 

1978

Sakai et al., 1985 Gibson et al., 

1978

Kaden et al., 

1979

MerschSundermann et 

al., 1992

Segerback and 

Vodicka, 1993

Cherng et al., 

2001

Value of RPF

Reference

RPF detection limits for 

nonpositive bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
142 DRAFT – DO NOT CITE OR QUOTE 

1 Benz[j]aceanthrylene (BjAC) 

2 

3 

4 

5 Benz[j]aceanthrylene (CASRN 202-33-5) is a nonalternant PAH comprised of four 

6 aromatic rings and one five-membered ring. Benz[j]aceanthrylene contains a classic bay region 

7 but no fjord region in its structure. 

8 There were 12 datasets for benz[j]aceanthrylene that met selection criteria and included 

9 benzo[a]pyrene (Figure 6-13); all of the studies gave positive results. The database includes one 

10 in vivo tumor bioassay dataset, one in vivo DNA adduct dataset, four mutagenicity or 

11 morphological/malignant cell transformation datasets, and six in vitro DNA damage or DNA 

12 adduct datasets. In a bioassay of benz[j]aceanthrylene that used intraperitoneal injection in an 

13 A/J mouse system (Mass et al., 1993), all mice treated with benz[j]aceanthrylene developed 

14 tumors (incidence of 100% at doses of 20–100 mg/kg; incidence for benzo[a]pyrene was 63–

15 100% across the same dose range), precluding the derivation of an RPF using incidence data. 

16 However, tumor multiplicity (average number of tumors per animal) data were available for 

17 dose-response modeling and resulted in an RPF estimate of 60. Benz[j]aceanthrylene treatment 

18 resulted in a pronounced increase in the average number of tumors per animal (59.45 tumors per 

19 animal at 20 mg/kg), much higher than benzo[a]pyrene treatment (5.05 tumors per animal at 

20 100 mg/kg), indicating that this compound is very potent in this test system. In a dermal 

21 initiation bioassay that did not include benzo[a]pyrene, benz[j]aceanthrylene induced papillomas 

22 in 90% of mice treated with an initiating dose of 40 μg (compared with 5% incidence in 

23 controls). As the available bioassay that included benzo[a]pyrene was positive and suggested 

24 that this compound is very potent, benz[j]aceanthrylene was considered potentially carcinogenic 

25 and was selected for inclusion in the RPF approach.

26 
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1 2 Figure 6-13. Benz[j]aceanthrylene (BjAC) RPFs.

0.001

0.01

0.1

1

10

100

Mass et al., 

1993

Mass et al., 

1993

Johnsen et al., 

1997

Johnsen et al., 

1997

Johnsen et al., 

1997

Sangaiah et 

al., 1983

Nesnow et al., 

1984

Mohapatra et 

al., 1987

Johnsen et al., 

1998

Johnsen et al., 

1998

Johnsen et al., 

1998

Johnsen et al., 

1998

Value RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
144 DRAFT – DO NOT CITE OR QUOTE 

1 Benzo[j]fluoranthene (BjF) 

2 

3 Benzo[j]fluoranthene (CASRN 205-82-3) is a nonalternant PAH comprised of four 

4 aromatic rings and one five-membered ring. Benzo[j]fluoranthene does not contain a classic bay 

5 or fjord region in its structure. 

6 There were eight datasets for benzo[j]fluoranthene that met selection criteria and 

7 included benzo[a]pyrene (Figure 6-14): seven in vivo tumor bioassay datasets and one in vitro 

8 study of DNA damage. Of the seven bioassay datasets, significant increases in tumor incidence 

9 or count were observed in all but one. Significant increases in tumor incidence were reported in 

10 both male and female mice tested in a newborn mouse bioassay using intraperitoneal injection of 

11 single doses (LaVoie et al., 1987), a mouse dermal initiation study (LaVoie et al., 1982), and a 

12 rat lung implantation bioassay (Deutsch-Wenzel et al., 1983). Significant increases in tumor 

13 multiplicity were reported in two mouse dermal initiation studies (Weyand et al., 1992; LaVoie 

14 et al., 1982). The one nonpositive bioassay was a mouse dermal complete carcinogenicity 

15 bioassay with an RPF detection limit of 0.1 (Habs et al., 1980). The in vitro study of DNA 

16 damage gave positive results (Mersch-Sundermann et al., 1992). Because the inconsistent 

17 bioassay results can be attributed to different test systems or study design, benzo[j]fluroanthene 

18 was considered potentially carcinogenic and was selected for inclusion in the RPF approach. 

19 
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1 2 Figure 6-14. Benzo[j]fluoranthene (BjF) RPFs. 

0.001

0.01

0.1

1

10

100

LaVoie et al., 1987 LaVoie et al., 1987 LaVoie et al., 1982 LaVoie et al., 1982 Deutsch-Wenzel et 

al., 1983

Weyand et al., 1992 Habs et al., 1980 MerschSundermann et al

1992

Value of RPF

Reference

RPF detection 

limit for 

nonpositive 

bioassay

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
146 DRAFT – DO NOT CITE OR QUOTE 

1 Benzo[k]fluoranthene (BkF) 

2 

3 

4 

5 Benzo[k]fluoranthene (CASRN 207-08-9) is a nonalternant PAH comprised of four 

6 aromatic rings and one five-membered ring. Benzo[j]fluoranthene does not contain a classic bay 

7 or fjord region in its structure. 

8 There were five datasets for benzo[k]fluoranthene that met selection criteria and included 

9 benzo[a]pyrene (Figure 6-15). The database includes four in vivo tumor bioassay datasets and 

10 one morphological/malignant cell transformation dataset. Statistically significant increases in 

11 tumor incidence and tumor count were reported in a mouse dermal initiation study (LaVoie et al., 

12 1982) and increased tumor incidence was reported in a rat lung implantation bioassay (Deutsch13 Wenzel et al., 1983). No significant increase in tumor incidence was observed in a dermal 

14 complete carcinogenicity study with an RPF detection limit of 0.1 (Habs et al., 1980). The 

15 morphological/malignant cell transformation study (Emura et al., 1980) was negative. Because 

16 the inconsistent bioassay results can be attributed to different test systems or study design 

17 (dermal initiation vs. dermal complete carcinogenicity), benzo[k]fluroanthene was considered 

18 potentially carcinogenic and was selected for inclusion in the RPF approach. 

19 
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1 2 Figure 6-15. Benzo[k]fluoranthene (BkF) RPFs*.

0.001

0.01

0.1

1

10

100

Deutsch-Wenzel et al., 

1983

LaVoie et al., 1982 LaVoie et al., 1982 Habs et al., 1980 Emura et al., 1980

Value of RPF

Reference

RPF detection limits for 

nonpositive bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
148 DRAFT – DO NOT CITE OR QUOTE 

1 Benz[l]aceanthrylene (BlAC) 

2 

3 

4 Benz[l]aceanthrylene (CASRN 211-91-6) is a nonalternant PAH comprised of four 

5 aromatic rings and one five-membered ring. Benz[l]aceanthrylene does not contain a classic bay 

6 or fjord region in its structure. 

7 There were 16 datasets for benz[l]aceanthrylene that met selection criteria and included 

8 benzo[a]pyrene (Figure 6-16); all of the studies gave positive results. The database includes four 

9 in vivo tumor bioassay datasets, five mutagenicity or morphological/malignant cell 

10 transformation datasets, one in vivo clastogenicity dataset, and six in vitro DNA adduct or DNA 

11 damage datasets. Significant increases in tumor count and multiplicity were reported in both 

12 male and female mice in a dermal initiation bioassay (Nesnow et al., 1984). All of the cancer13 related endpoint studies were positive as well. Relative potency estimates for most of the 

14 available datasets were ≥1.0, suggesting equivalent or greater potency than benzo[a]pyrene. As 

15 the available bioassays that included benzo[a]pyrene were positive, benz[l]aceanthrylene was 

16 considered potentially carcinogenic and was selected for inclusion in the RPF approach. 
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1 2 Figure 6-16. Benz[l]aceanthrylene (BlAC) RPFs.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
150 DRAFT – DO NOT CITE OR QUOTE 

1 Chrysene (CH) 

2 

3 

4 Chrysene (CASRN 218-01-9) is an alternant PAH comprised of four fused aromatic 

5 rings. Chrysene contains two bay regions but no fjord region in its structure. 

6 There were 40 datasets for chrysene that met selection criteria and included 

7 benzo[a]pyrene (Figure 6-17). Included in the database are 13 in vivo tumor bioassay datasets, 

8 4 in vivo DNA adduct datasets, 3 in vivo clastogenicity datasets, 11 mutagenicity datasets, 

9 3 morphological/malignant cell transformation datasets, and 6 in vitro studies of DNA damage, 

10 adducts, or clastogenicity. Among the bioassays that included benzo[a]pyrene, 11 reported 

11 significant increases in tumor incidence or tumor multiplicity, and 3 did not. Significant 

12 increases in tumor incidence and/or multiplicity were reported in three dermal initiation studies 

13 in mice (Rice et al., 1988; Slaga et al., 1980; Hecht et al., 1974), a newborn mouse study in 

14 males (Wislocki et al., 1986), and a rat lung implantation bioassay (Wenzel-Hartung et al., 

15 1990). Female mice tested in the newborn mouse assay published by Wislocki et al. (1986) did 

16 not have a significant increase in tumor incidence, resulting in one of the three nonpositive 

17 studies. The other two nonpositive findings were in males and females tested in another 

18 newborn mouse bioassay (Busby et al., 1989). The bioassays with nonpositive findings had RPF 

19 detection limits between 0.06 and 0.2. Conflicting results in male mice were reported in the two 

20 newborn mouse bioassays (Busby et al., 1989; Wislocki et al., 1986). The major difference 

21 between the two studies is the duration of follow-up; Busby et al. (1989) sacrificed the mice at 

22 26 weeks, while Wislocki et al. (1986) followed the mice for a full year. LaVoie et al. (1994) 

23 observed that liver tumor induction in the newborn mouse bioassay is not fully realized until the 

24 mice have reached 1 year of age, and the positive findings by Wislocki et al. (1986) indeed 

25 reflect liver tumors in the male mice. Chrysene was shown to form DNA adducts when 

26 administered in vivo in both rats and mice via injection and gavage (Kligerman et al., 2002). 

27 Bacterial and mammalian mutagenicity and morphological/malignant cell transformation assays 

28 of chrysene were all positive, as were studies of clastogenicity tested in vivo. In contrast, results

29 from in vitro studies of DNA adducts, DNA damage, and clastogenicity were not consistent. 

30 Because the inconsistent bioassay results can be attributed to different study designs 

31 (gender, follow-up time), chrysene was considered potentially carcinogenic and was selected for 

32 inclusion in the RPF approach. 

33 
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1 2 Figure 6-17. Chrysene (CH) RPFs*.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

RPF detection limits for nonpositive bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Coronene (CO) 

2 

3 Coronene (CASRN 191-07-1) is an alternant PAH comprised of seven fused aromatic 

4 rings. Coronene contains no bay or fjord regions in its structure. 

5 There were six datasets for coronene that met selection criteria and included 

6 benzo[a]pyrene (Figure 6-18). A dermal complete carcinogenicity bioassay in mice did not 

7 result in a statistically significant increase in tumor incidence (Habs et al., 1980); the RPF 

8 detection limit was 0.06. To confirm the nonpositive findings in the one tumor bioassay that 

9 included benzo[a]pyrene, other bioassays and cancer-related endpoint data were considered. 

10 There was one bioassay of coronene that did not include benzo[a]pyrene. Van Duuren et al. 

11 (1968) conducted a dermal initiation bioassay of coronene using groups of 20 mice (0.5 mg 

12 coronene in 0.5 mL benzene, followed by croton resin treatment until death). Although the 

13 authors characterized coronene as a weak tumor initiator, the incidence of tumors was not 

14 significantly increased over concurrent controls. The limited cancer-related endpoint data were 

15 mixed, with three positive bacterial mutagenicity studies (with RPFs ranging from 0.01 to 0.5), 

16 one negative bacterial mutagenicity study, and a negative in vitro DNA damage study. 

17 Overall, the database for coronene is both limited and inconsistent. Because the database 

18 for coronene does not provide adequate information with which to assess potential 

19 carcinogenicity, this PAH was not selected for inclusion in the RPF approach. 
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1 2 Figure 6-18. Coronene (CO) RPFs*.

0.001

0.01

0.1

1

10

100

Habs et al., 1980 Hermann, 1981 Sakai et al., 1985 Florin et al., 1980 Kaden et al., 1979 Mersch-Sundermann 

al., 1992

Value of RPF

Reference

RPF detection limits for 

nonpositive bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Cyclopenta[c,d]pyrene (CPcdP) 

2 

3 Cyclopenta[c,d]pyrene (CASRN 27208-37-3) is a nonalternant PAH comprised of four 

4 aromatic rings and one five-membered ring. Cyclopenta[c,d]pyrene does not contain a classic 

5 bay or fjord region in its structure. 

6 There were 24 datasets for cyclopenta[c,d]pyrene that met selection criteria and included 

7 benzo[a]pyrene (Figure 6-19). The database includes 10 in vivo tumor bioassay datasets, 2 in 

8 vivo DNA adduct datasets, 11 studies of mutagenicity or morphological/malignant cell 

9 transformation, and a single study of in vitro clastogenicity. Eight of the 10 tumor bioassay 

10 datasets and all of the cancer-related endpoint studies gave positive results. Statistically 

11 significant increases in tumor incidence and/or multiplicity were reported in two dermal 

12 complete carcinogenicity bioassay (Cavalieri et al., 1983, 1981b), two dermal initiation 

13 bioassays (Raveh et al., 1982; Cavalieri et al., 1981b), and an intraperitoneal study using adult 

14 A/J mice (Nesnow et al., 1998b). Bioassays in which no significant increase in tumorigenicity 

15 was observed included a dermal initiation (Wood et al., 1980) and complete carcinogenicity 

16 study (Habs et al., 1980); these studies had RPF detection limits of 0.1 and 0.03, respectively. 

17 After obtaining nonpositive results for low initiating doses of cyclopenta[c,d]pyrene, Wood et al. 

18 (1980) repeated their experiment with higher doses and observed statistically significant 

19 increases in tumor incidence. In the latter experiment, benzo[a]pyrene was not included, so an 

20 RPF could not be calculated from these data. The study design of the nonpositive complete 

21 carcinogenicity bioassay was quite similar to that of the two positive studies of this type, with the 

22 exception of the mouse strain used; Habs et al. (1980) used NMRI mice, while Cavalieri et al. 

23 (1983, 1981b) used Swiss mice. Although the differing results in dermal complete 

24 carcinogenicity studies may be explained by slight differences in strain susceptibility, these two 

25 strains are of common origin, which argues against this explanation. 

26 The available cancer-related endpoint data indicate that cyclopenta[c,d]pyrene is 

27 mutagenic and capable of morphological/malignant cell transformation in vitro; a single study of 

28 in vitro clastogenicity was also positive. Overall, the data supporting a finding of potential 

29 carcinogenicity for cyclopenta[c,d]pyrene are very consistent, and this compound was selected 

30 for inclusion in the RPF approach. 
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1 2 Figure 6-19. Cyclopenta[c,d]pyrene (CPcdP) RPFs.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

RPF detection limits for 

nonpositive bioassays

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 4H-Cyclopenta[d,e,f]chrysene (CPdefC) 

2 

3 

4 4H-Cyclopenta[d,e,f]chrysene (CASRN 202-98-2) is a nonalternant PAH comprised of 

5 four aromatic rings and one five-membered ring. 4H-Cyclopenta[d,e,f]chrysene contains a 

6 classic bay region but no fjord region in its structure. 

7 There were two datasets for 4H-cyclopenta[d,e,f]chrysene that met selection criteria and 

8 included benzo[a]pyrene (Figure 6-20): both were multi-dose dermal initiation datasets (Rice et 

9 al., 1988, 1985). Rice et al. (1988) reported a statistically significant increase in tumor incidence

10 in a multi-dose dermal initiation study. In the second study, the incidence of tumors after 

11 treatment with cyclopenta[d,e,f]chrysene exceeded 90%, precluding RPF derivation from 

12 incidence data, but tumor multiplicity data were available for RPF calculation (Rice et al., 1985). 

13 Cyclopenta[d,e,f]chrysene has not been tested in a bioassay without benzo[a]pyrene; however, 

14 sterically hindered diol epoxides of this compound have given positive results in a newborn 

15 mouse assay (Amin et al., 1995). Because the bioassay of cyclopenta[d,e,f]chrysene was 

16 positive, this PAH was considered potentially carcinogenic and was selected for inclusion in the 

17 RPF approach.
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1 2 Figure 6-20. Cyclopenta[d,e,f]chrysene (CPdefC) RPFs.

0.001

0.01

0.1

1

10

100

Rice et al., 1988 Rice et al., 1985

Value of RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Dibenz[a,c]anthracene (DBacA) 

2 

3 

4 Dibenz[a,c]anthracene (CASRN 215-58-7) is an alternant PAH comprised of five fused 

5 aromatic rings. Dibenz[a,c]anthracene contains three bay regions but no fjord region in its 

6 structure. 

7 There were 15 datasets for dibenz[a,c]anthracene that met selection criteria and included 

8 benzo[a]pyrene (Figure 6-21). The database includes a single in vivo study of DNA adducts, 

9 nine mutagenicity or morphological/malignant cell transformation studies, and five studies of in 

10 vitro DNA damage or adducts. One morphological/malignant cell transformation assay gave 

11 nonpositive results, while the remaining studies were positive. In the absence of positive 

12 bioassays with benzo[a]pyrene, other bioassays and cancer-related data were considered to 

13 evaluate the potential carcinogenicity of dibenz[a,c]anthracene. 

14 Conflicting results were reported in three dermal initiation bioassays of 

15 dibenz[a,c]anthracene in which benzo[a]pyrene was not included. Van Duuren et al. (1970) 

16 observed a tumor incidence of 95% (19/20, compared to 1/20 controls) when mice were treated 

17 with an initiating dose of 1 mg dibenz[a,c]anthracene in benzene followed by thrice weekly 

18 treatment with phorbol myristate acetate. In contrast, there was no significant increase in tumor 

19 formation when the same initiating dose was followed by thrice weekly application of croton 

20 resin (Van Duuren et al., 1968); however, the latency to first tumor was substantially reduced 

21 (65 vs. 150 days in controls). Latency was also substantially reduced in the study by Van 

22 Duuren et al. (1970), in which the first tumor appeared after 74 days, compared with 338 days in 

23 controls. 

24 Cancer-related endpoint data for dibenz[a,c]anthracene are predominantly positive 

25 (8/9 mutagenicity or morphological/malignant cell transformation studies and 5/5 studies of in 

26 vitro DNA adducts or DNA damage). Although the conflicting bioassay data are not easily 

27 explained, the high incidence of tumors (19/20) in the study by Van Duuren et al. (1970), and the 

28 reduced latency to tumor formation in both studies, coupled with predominantly positive cancer29 related endpoint data, suggest that dibenz[a,c]anthracene is potentially carcinogenic. 

30 Contributing to this conclusion is the observation that dibenz[a,c]anthracene is an alternant PAH 

31 with known structural alerts for carcinogenicity (more than three rings, and three bay regions). 

32 Thus, dibenz[a,c]anthracene was selected for inclusion in the RPF approach. 
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1 2 Figure 6-21. Dibenz[a,c]anthracene (DBacA) RPFs*.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference * Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
160 DRAFT – DO NOT CITE OR QUOTE 

1 Dibenzo[a,e]fluoranthene (DBaeF) 

2 

3 Dibenzo[a,e]fluoranthene (CASRN 5385-75-1) is a nonalternant PAH comprised of five 

4 aromatic rings and one five-membered ring. Dibenzo[a,e]fluoranthene contains a classic bay 

5 region but no fjord region in its structure. 

6 There were three datasets for dibenzo[a,e]fluoranthene that met selection criteria and 

7 included benzo[a]pyrene (Figure 6-22); all gave positive results. The database includes two in 

8 vivo tumor bioassays and one mammalian mutagenicity study. Statistically significant increases 

9 in tumor incidence were reported in dermal initiation and complete carcinogenicity bioassays in 

10 mice (both reported by Hoffmann and Wynder, 1966). As the available bioassays for 

11 dibenzo[a,e]fluoranthene were positive, this compound was considered potentially carcinogenic 

12 and was selected for inclusion in the RPF approach.

13 
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1 2 Figure 6-22. Dibenzo[a,e]fluoranthene (DBaeF) RPFs.

0.001

0.01

0.1

1

10

100

Hoffmann and Wynder 1966 Hoffmann and Wynder 1966 Durant et al., 1999

Value of RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Dibenzo[a,e]pyrene (DBaeP) 

2 

3 

4 Dibenzo[a,e]pyrene (CASRN 192-65-4) is an alternant PAH comprised of six fused 

5 aromatic rings. Dibenzo[a,e]pyrene contains three bay regions but no fjord region in its 

6 structure. 

7 There were three datasets for dibenzo[a,e]pyrene that met selection criteria and included 

8 benzo[a]pyrene (Figure 6-23). The database includes two in vivo tumor bioassay datasets and 

9 one in vitro bacterial mutagenicity dataset, all of which gave positive results. Statistically 

10 significant increases in tumor incidence were reported in dermal initiation and complete 

11 carcinogenicity bioassays in mice (Hoffmann and Wynder, 1966). The complete carcinogenicity 

12 bioassay was confounded by significant toxicity-related mortality unrelated to tumors (Hoffmann 

13 and Wynder, 1966). The one bacterial mutagenicity study reported positive results. Because the 

14 available bioassays with benzo[a]pyrene were both positive, dibenzo[a,e]pyrene was considered 

15 potentially carcinogenic and was selected for inclusion in the RPF approach. 

16 
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1 2 Figure 6-23. Dibenzo[a,e]pyrene (DBaeP) RPFs.

0.001

0.01

0.1

1

10

100

Hoffmann and Wynder 1966 Hoffmann and Wynder 1966 Teranishi et al., 1975

Value of RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Dibenz[a,h]anthracene (DBahA) 

2 

3 Dibenz[a,h]anthracene (CASRN 53-70-3) is an alternant PAH comprised of five fused 

4 aromatic rings. Dibenz[a,h]anthracene contains two bay regions and no fjord region in its 

5 structure. 

6 There were 30 datasets for dibenz[a,h]anthracene that met selection criteria and included 

7 benzo[a]pyrene (Figure 6-24). Included in the database are in vivo tumor bioassay datasets (4), 

8 in vivo DNA adduct datasets (2), an in vivo clastogenicity dataset, mutagenicity datasets (10) 

9 morphological/malignant cell transformation datasets (6), and in vitro DNA damage, adducts, or 

10 clastogenicity datasets (7). There were three tumor bioassays for dibenz[a,h]anthracene that 

11 included benzo[a]pyrene, and all resulted in statistically significant increases in tumor incidence 

12 and/or multiplicity. The bioassays were in three different test systems: a rat lung implantation 

13 study (Wenzel-Hartung et al., 1990), a mouse dermal initiation study reporting both incidence 

14 and multiplicity (Slaga et al., 1980), and an intraperitoneal study in A/J mice (Nesnow et al., 

15 1998b). Dibenz[a,h]anthracene was shown to form DNA adducts when administered in vivo to 

16 mice via intraperitoneal injection (Nesnow et al., 1998b) and dermal application (Phillips et al., 

17 1979). Mutagenicity and morphological/malignant cell transformation assays of 

18 dibenz[a,h]anthracene were predominantly positive (13/16), as were studies of other cancer19 related endpoints. Because the available bioassays with benzo[a]pyrene were positive, 

20 dibenz[a,h]anthracene was considered potentially carcinogenic and was selected for inclusion in 

21 the RPF approach. 

22 
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1 2 Figure 6-24. Dibenz[a,h]anthracene (DBahA) RPFs*.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference * Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Dibenzo[a,h]pyrene (DBahP) 

2 

3 Dibenzo[a,h]pyrene (CASRN 189-64-0) is an alternant PAH comprised of six fused 

4 aromatic rings. Dibenzo[a,h]pyrene contains two bay regions and no fjord region in its structure. 

5 There were five datasets for dibenzo[a,h]pyrene that met selection criteria and included 

6 benzo[a]pyrene (Figure 6-25); all gave positive results. The database includes one in vivo 

7 bioassay dataset, one in vivo DNA adduct dataset, two in vitro mammalian mutagenicity 

8 datasets, and one in vitro DNA damage dataset. A statistically significant increase in tumor 

9 incidence was reported in a dermal initiation bioassay in mice (Hoffmann and Wynder, 1966). 

10 In addition, two dermal studies of complete carcinogenicity that included benzo[a]pyrene gave 

11 positive results, but no RPF could be calculated because the incidence of tumors in the mice 

12 exposed to dibenzo[a,h]pyrene was ≥90% at the lowest dose tested (Cavalieri et al., 1977; 

13 Hoffmann and Wynder, 1966) and tumor multiplicity was not reported. As all of the available 

14 bioassays that included benzo[a]pyrene showed exposure-related tumorigenic responses, 

15 dibenzo[a,h]pyrene was considered potentially carcinogenic and was selected for inclusion in the 

16 RPF approach. 

17 
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1 2 Figure 6-25. Dibenzo[a,h]pyrene (DBahP) RPFs.

0.001

0.01

0.1

1

10

100

Hoffmann and Wynder 1966 Hughes and Phillips 1990 Durant et al., 1999 Hass et al., 1982 Mersch-Sundermann et a

1992

Value of RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Dibenzo[a,i]pyrene (DBaiP) 

2 

3 Dibenzo[a,i]pyrene (CASRN 189-55-9) is an alternant PAH comprised of six fused 

4 aromatic rings. Dibenzo[a,i]pyrene contains two bay regions and no fjord region in its structure. 

5 There were 12 datasets for dibenzo[a,i]pyrene that met selection criteria and included 

6 benzo[a]pyrene (Figure 6-26); all gave positive results. The database includes two in vivo 

7 bioassay datasets, one in vivo DNA adduct dataset, seven in vitro mutagenicity datasets, and two 

8 in vitro DNA damage datasets. Statistically significant increases in tumor incidence were 

9 reported in dermal initiation and complete carcinogenicity bioassays in mice, both published by 

10 Hoffmann and Wynder (1966). The cancer-related endpoint studies were all positive. As the 

11 available bioassays that included benzo[a]pyrene were both positive, dibenzo[a,i]pyrene was 

12 considered potentially carcinogenic and was selected for inclusion in the RPF approach. 

13 
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1 2 Figure 6-26. Dibenzo[a,i]pyrene (DbaiP) RPFs*.

0.001

0.01

0.1

1

10

100

Hoffmann and 

Wynder 1966

Hoffmann and 

Wynder 1966

Hughes and 

Phillips 1990

Durant et al., 

1999

Hass et al., 

1982

Teranishi et 

al., 1975

Hermann, 

1981

MerschSundermann et 

al., 1992)

Phillipson and 

Ioannides 1989

Baker et al., 

1980

Ichinotsubo et 

al., 1977

McCann e

1975

Value of RPF

Reference * Missing bar indicates nonpositive genotoxicity study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Dibenzo[a,l]pyrene (DBalP). 

2 

3 Dibenzo[a,l]pyrene (CASRN 191-30-0) is an alternant PAH comprised of six fused 

4 aromatic rings. Dibenzo[a,l]pyrene contains both a bay region and a fjord region in its structure. 

5 There were 15 datasets for dibenzo[a,l]pyrene that met selection criteria and included 

6 benzo[a]pyrene (Figure 6-27); all of the studies gave positive results. The database includes 

7 three in vivo tumor bioassay datasets, three in vivo DNA adduct datasets, one bacterial 

8 mutagenicity dataset, one morphological/malignant cell transformation dataset, four in vivo 

9 clastogenicity datasets, and three in vitro DNA adduct or DNA damage datasets. 

10 In three bioassays of dibenzo[a,l]pyrene included benzo[a]pyrene, RPFs could not be 

11 calculated using incidence data, because the incidence of tumors associated with the lowest dose 

12 of dibenzo[a,l]pyrene exceeded 90% (two dermal initiation experiments in mice and an 

13 intramammilary injection study in rats, both reported by Cavalieri et al., 1991); however, tumor 

14 multiplicity data were reported for the dermal initiation experiments and were used to calculate 

15 RPFs of 10 and 40. Nesnow et al. (1998b) reported tumor multiplicity, but not tumor incidence 

16 in A/J mice exposed intraperitoneally; an RPF of 30 was calculated. Because the available 

17 studies indicated that dibenzo[a,l]pyrene may be much more potent benzo[a]pyrene, other studies 

18 were also examined to confirm the potency of this compound. 

19 Dibenzo[a,l]pyrene treatment resulted in significant increases in tumor incidence in seven 

20 bioassays that did not include benzo[a]pyrene, including two dermal initiation studies (Gill et al., 

21 1994; Cavalieri et al., 1989), a dermal complete carcinogenicity study (Nakatsuru et al., 2004), 

22 an intramammilary injection study in rats (Cavalieri et al., 1989), a newborn mouse bioassay 

23 (Platt et al., 2004), an intraperitoneal bioassay using A/J mice (Prahalad et al., 1997), and a 

24 gavage bioassay comparing the responses of cyp1B1 wild-type and null mice (Buters et al., 

25 2002). In several of these studies, there was significant toxicity associated with 

26 dibenzo[a,l]pyrene treatment. Tumor incidences were very high in most of the studies, including 

27 the gavage study (Buters et al., 2002), which reported an overall tumor incidence of 100% in 

28 cyp1B1 wild-type mice treated with a single dose of dibenzo[a,l]pyrene. A recent study 

29 examining in utero and/or lactational exposure to dibenzo[a,l]pyrene showed that mouse pups 

30 exposed during late gestation develop T-cell lymphomas between 3 and 6 months of age, as well 

31 multiple lung and liver tumors (Castro et al., 2008). All of the cancer-related data for 

32 dibenzo[a,l]pyrene were positive and resulted in high RPF estimates, including in vivo and in 

33 vitro studies of DNA adducts, in vivo clastogenicity studies, morphological/malignant cell 

34 transformation, bacterial mutagenicity, and in vitro DNA damage or DNA adduct studies. 
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1 The weight of evidence supporting a finding of potential carcinogenicity for 

2 dibenzo[a,l]pyrene is strong and suggests that this compound is very potent; thus, it was selected 

3 for inclusion in the RPF approach. 

4 
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1 2 Figure 6-27. Dibenzo[a,l]pyrene (DBalP) RPFs.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Fluoranthene (FA) 

2 

3 Fluoranthene (CASRN 206-44-0) is a nonalternant PAH comprised of three aromatic 

4 rings and one five-membered ring. Fluoranthene does not contain a classic bay or fjord region in 

5 its structure. 

6 There were 21 datasets for fluoranthene that met selection criteria and included 

7 benzo[a]pyrene (Figure 6-28). Included in the database are in vivo tumor bioassay datasets (11), 

8 bacterial and mammalian mutagenicity datasets (5), a morphological/malignant cell 

9 transformation assay, and in vitro studies of DNA damage, DNA adducts, or clastogenicity (4). 

10 Of the bioassay datasets that included benzo[a]pyrene, nine gave positive results and two gave 

11 nonpositive results. Statistically significant increases in tumor incidence and tumor multiplicity 

12 were reported in newborn mouse bioassays (in male and female mice [LaVoie et al., 1994] and in 

13 female mice [Busby et al., 1989]). The tumor incidence was not significantly increased by 

14 fluoranthene in a mouse dermal initiation study with an RPF detection limit of 0.01 (Hoffman et 

15 al., 1972) and when fluoranthene was tested alone in a dermal cocarcinogenicity bioassay with 

16 an RPF detection limit of 0.1 (Van Duuren and Goldschmidt, 1976). In another newborn mouse 

17 bioassay (Busby et al., 1984) that reported both incidence and multiplicity, the lowest dose of 

18 benzo[a]pyrene resulted in a tumor incidence of >90%, precluding RPF calculation from the 

19 incidence data; however, multiplicity data were available. Statistical analysis of the data for 

20 fluoranthene demonstrated positive findings for both incidence and multiplicity in male mice, but 

21 the results for the two endpoints were inconsistent in females. In female mice exposed at the 

22 high dose of fluoranthene in a newborn mouse bioassay reported by Busby et al. (1984), the lung 

23 tumor count was significantly increased (albeit borderline, p = 0.0343) while the incidence was 

24 not (p > 0.05), and neither was statistically significantly increased at the lower dose. For the 

25 purpose of this analysis, the multiplicity data were treated as an independent measure of 

26 carcinogenic potency, and an RPF was calculated for the statistically increased tumor count in 

27 female mice. 

28 The mutagenicity studies of fluoranthene were all positive, but in vitro studies of DNA 

29 damage, DNA adducts, and clastogenicity gave inconsistent results. Because the inconsistent 

30 bioassay results can be attributed to different test systems (different exposure route and/or 

31 gender) or study design, fluoranthene was considered potentially carcinogenic and was selected 

32 for inclusion in the RPF approach. 

33 
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1 2 Figure 6-28. Fluoranthene (FA) RPFs*.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

RPF detection limits for 

nonpositive bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Fluorene (FE) 

2 

3 Fluorene (CASRN 86-73-7) is a nonalternant PAH comprised of two aromatic rings and 

4 one five-membered ring. Fluorene does not contain a classic bay or fjord region in its structure. 

5 There were nine datasets for fluorene that met selection criteria and included 

6 benzo[a]pyrene (Figure 6-29). There were no tumor bioassays of fluorene that included 

7 benzo[a]pyrene, so other bioassays and cancer-related endpoint data were considered. LaVoie et 

8 al. (1980) conducted a study of skin tumor initiation in mice treated with 1.0 mg fluorene 

9 followed by 20 weeks of treatment with TPA; the study did not include benzo[a]pyrene. The 

10 incidence of tumor-bearing animals (5%) was not significantly increased over controls (0%) 

11 (LaVoie et al.,1980). The limited cancer-related endpoint data were mixed, with three positive 

12 and four negative mutagenicity datasets, and two negative in vitro DNA damage datasets. 

13 Overall, the database for fluorene is both limited and inconsistent. Because the database for 

14 fluorene does not provide adequate information with which to assess potential carcinogenicity, 

15 this PAH was not selected for inclusion in the RPF approach. 

16 

17 
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1 2 Figure 6-29. Fluorene (FE) RPFs*.

0.001

0.01

0.1

1

10

100

Sakai et al., 1985 Gibson et al., 1978 Wangenheim and

Bolcsfoldi 1988

Hermann, 1981 Kaden et al., 1979 McCann et al., 

1975

Pahlman and 

Pelkonen, 1987

Mamber et al., 

1983

MerschSundermann et a

1992

Value of RPF

* Missing bar indicates nonpositive cancer-related endpoint study Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Indeno[1,2,3-c,d]pyrene (IP) 

2 

3 Indeno[1,2,3-c,d]pyrene (CASRN 193-39-5) is a nonalternant PAH comprised of five 

4 aromatic rings and one five-membered ring. Indeno[1,2,3-c,d]pyrene does not contain a classic 

5 bay or fjord region in its structure. 

6 There were five datasets for indeno[1,2,3-c,d]pyrene that met selection criteria and 

7 included benzo[a]pyrene (Figure 6-30). There are three tumor bioassays, one in vitro study of 

8 morphological/malignant cell transformation (Emura et al., 1980), and one in vitro study of DNA 

9 damage (Mersch-Sundermann et al., 1992). Of the three tumor bioassays, only one, a rat lung 

10 implantation study (Deutsch-Wenzel et al., 1983), reported a statistically significant increase in 

11 tumor incidence or multiplicity; the RPF was 0.07. Nonpositive findings were reported in mouse 

12 dermal initiation (Hoffmann and Wyner, 1966) and complete carcinogenicity (Habs et al., 1980) 

13 studies with RPF detection limits in the range of 0.1–0.3. Because the inconsistent bioassay 

14 results can be attributed to different test systems (different species and route), and the 

15 nonpositive studies may not have been sufficiently sensitive to detect an effect, indeno[1,2,3-c,d]

16 pyrene was considered potentially carcinogenic and was selected for inclusion in the RPF 

17 approach. 

18 
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1 2 Figure 6-30. Indeno[1,2,3-c,d]pyrene (IP) RPFs.

0.001

0.01

0.1

1

10

100

Deutsch-Wenzel et al., 1983 Hoffmann and Wynder 1966 Habs et al., 1980 Mersch-Sundermann et al., 1992 Emura et al., 1980

Value of RPF

Reference

RPF detection limits for 

nonpositive bioassays

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Naphtho[2,3-e]pyrene (N23eP) 

2 

3 Naphtho[2,3-e]pyrene (CASRN 193-09-9) is an alternant PAH comprised of six fused 

4 aromatic rings. Naphtho[2,3-e]contains two bay regions and no fjord region in its structure. 

5 There were two datasets for naphtho[2,3-e]pyrene that met selection criteria and included 

6 benzo[a]pyrene (Figure 6-31): a tumor bioassay dataset and an in vitro mammalian mutagenicity 

7 dataset (both were positive). The tumor bioassay was a single dose dermal initiation bioassay 

8 (Hoffmann and Wynder, 1966). As the available bioassay reported a statistically significant 

9 increase in tumor incidence, naphtho[2,3-e]pyrene was considered potentially carcinogenic, and 

10 was selected for inclusion in the RPF approach. 

11 
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1 2 Figure 6-31. Naphtho[2,3-e]pyrene (N23eP) RPFs.

0.001

0.01

0.1

1

10

100

Hoffmann and Wynder 1966 Durant et al., 1999

Value of RPF

Reference

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Perylene (Pery) 

2 

3 Perylene (CASRN 198-55-0) is an alternant PAH comprised of five fused aromatic rings. 

4 Perylene contains two bay regions and no fjord region in its structure. 

5 There were 11 datasets for perylene that met selection criteria and included 

6 benzo[a]pyrene (Figure 6-32). The database includes an in vivo tumor bioassay dataset, an in 

7 vivo clastogenicity dataset, eight bacterial mutagenicity datasets, and an in vitro DNA damage 

8 dataset. The single tumor bioassay, a dermal initiation study, gave nonpositive results for 

9 perylene (El-Bayoumy et al., 1982); the RPF detection limit was 0.01. To confirm the 

10 nonpositive bioassay findings, other bioassays and cancer-related endpoint data were considered. 

11 In a study that did not include benzo[a]pyrene, Van Duuren et al. (1970) did not observe an 

12 increase in tumor incidence over controls when mice were treated by dermal application with an 

13 initiating dose of 0.8 mg perylene in benzene followed by thrice weekly treatment with phorbol 

14 myristate acetate for 58 weeks. However, seven of the eight bacterial mutagenicity studies gave 

15 positive results, while perylene tested negative in one bacterial mutagenicity study, the 

16 clastogenicity study, and the DNA damage study. Overall, the database for perylene is both 

17 limited and inconsistent. Because the database for perylene does not provide adequate 

18 information with which to assess potential carcinogenicity, this PAH was not selected for 

19 inclusion in the RPF approach. 

20 
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1 2 Figure 6-32. Perylene (Pery) RPFs*.

0.001

0.01

0.1

1

10

100

El-Bayoumy et 

al., 1982

Sirianni and 

Huang, 1978

Kaden et al., 

1979

Carver et al., 

1986

Sakai et al., 1985 Hermann, 1981 LaVoie et al., 

1979

De Flora et al., 

1984

Florin et al., 

1980

Pahlman and 

Pelkonen, 1987

Mersc

Sunderm

al., 19

Value of RPF

Reference

RPF detection limits for 

nonpositive bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Phenanthrene (PH) 

2 

3 Phenanthrene (CASRN 85-01-8) is an alternant PAH comprised of three fused aromatic 

4 rings. Phenanthrene contains a bay region in its structure, but has less than four aromatic rings. 

5 There were 34 datasets for phenanthrene that met selection criteria and included 

6 benzo[a]pyrene, including 3 in vivo tumor bioassay datasets, 2 in vivo clastogenicity datasets, 

7 11 mutagenicity datasets, 6 morphological/malignant cell transformation datasets, and 12 in vitro 

8 studies of DNA adducts, DNA damage, or clastogenicity (Figure 6-33). Only seven studies 

9 reported positive results; the remaining 27 studies reported nonpositive findings, including all 

10 three bioassays. Nonpositive findings were reported in the three bioassays that included 

11 benzo[a]pyrene, including a lung implantation study in rats (Wenzel-Hartung et al., 1990), a 

12 dermal initiation study in mice (LaVoie et al., 1981), and a subcutaneous study in mice (Grant 

13 and Roe, 1963). To confirm the nonpositive findings, other bioassays and cancer-related 

14 endpoint data were considered. In bioassays without benzo[a]pyrene, phenanthrene did not 

15 induce significant increases in tumors in a newborn mouse assay using a total dose of 1.4 μmol 

16 (Buening et al., 1979) or in two dermal initiation assays (Wood et al., 1979; Salaman and Roe, 

17 1956) using doses of 10 μmol and 540 mg, respectively. However, 12/30 mice developed 

18 papillomas by week 35 after dermal treatment with 10 μmol phenanthrene (in benzene) followed 

19 by twice weekly treatment with TPA; no control mice had papillomas (Scribner, 1973). The 

20 response was statistically significantly increased over controls (p < 0.01). 

21 In vitro assays of mutagenicity and morphological/malignant cell transformation were 

22 predominantly negative for phenanthrene. One of the two positive studies (Sakai et al., 1988) 

23 reported a poor dose-response relationship for phenanthrene. Two studies found evidence of 

24 clastogenicity after in vivo administration of phenanthrene (Roszinsky-Kocher et al., 1979; 

25 Bayer, 1978). However, in the study by Bayer (1978), only the high dose gave a significant 

26 response, and there was not a significant dose-response trend. When phenathrene was tested in 

27 in vitro studies of DNA adducts, DNA damage, and clastogenicity, the results were 

28 predominantly negative (9/12 studies). Overall, the database for phenanthrene is substantial, and 

29 the weight of evidence suggests that this PAH is not carcinogenic or is of very low carcinogenic 

30 potential. Based on the large number of negative bioassays and the abundant evidence that 

31 phenanthrene lacks genotoxic action, this compound was selected for inclusion in the RPF 

32 approach and assigned an RPF of 0. 

33 
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1 2 Figure 6-33. Phenanthrene (PH) RPFs*.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

RPF detection limits for 

nonpositive bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 Pyrene (Pyr) 

2 

3 

4 Pyrene (CASRN 129-00-0) is an alternant PAH comprised of four fused aromatic rings. 

5 Pyrene does not contain a bay or fjord region in its structure. 

6 There were 49 datasets for pyrene that met study quality criteria and included 

7 benzo[a]pyrene (Figure 6-34). Included in the database are in vivo tumor bioassay datasets (7), 

8 in vivo clastogenicity datasets (5), bacterial and mammalian mutagenicity datasets (14), 

9 morphological/malignant cell transformation datasets (7), and in vitro DNA damage, DNA 

10 adducts, or clastogenicity datasets (16). There were seven bioassays of pyrene that included 

11 benzo[a]pyrene; all gave nonpositive results. Nonpositive results were reported in two newborn 

12 mouse bioassays in which both males and females were tested (Busby et al., 1989; Wislocki et 

13 al., 1986), two studies of dermal initiation (El-Bayoumy et al., 1982; Wood et al., 1980), and a 

14 dermal cocarcinogenesis bioassay (Van Duuren and Goldschmidt, 1976). RPF detection limits in 

15 these studies ranged from about 0.01 to 0.1 (see Figure 6-34). In an intraperitoneal bioassay 

16 using A/J mice that included benzo[a]pyrene, the authors reported that pyrene treatment did not 

17 induce lung adenomas (Ross et al., 1995); data were not reported, so an RPF detection limit 

18 could not be estimated. In bioassays without benzo[a]pyrene, pyrene did not induce a significant 

19 increase in tumors in a dermal initiation bioassay (Salaman and Roe, 1956). Scribner (1973) 

20 reported a weak tumorigenic response in a dermal initiation study in mice (5/29 mice developed 

21 papillomas 35 weeks after dermal treatment with 10 μmol pyrene in benzene followed by twice 

22 weekly treatment with TPA as compared with 0/30 control mice, p = 0.02). 

23 In vitro assays of bacterial and mammalian mutagenicity and morphological/malignant 

24 cell transformation were predominantly negative for pyrene. In five studies of clastogenicity in 

25 animals exposed in vivo to pyrene, no evidence of clastogenic effects was reported. Further, in 

26 vitro studies of DNA adducts, DNA damage, and clastogenicity using pyrene also largely 

27 reported negative results. Overall, the database for pyrene is substantial, and the weight of 

28 evidence suggests that this PAH is not carcinogenic or is of very low carcinogenic potential. 

29 Based on the large number of negative bioassays and the abundant evidence that pyrene lacks 

30 genotoxic action, this compound was selected for inclusion in the RPF approach and assigned an 

31 RPF of 0. 

32 
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1 

2 Figure 6-34. Pyrene (Pyr) RPFs*.

0.001

0.01

0.1

1

10

100

Value of RPF

Reference

RPF detection limits for 

nonpositive bioassays

* Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicityIn vivo cancer-related endpoinIn vitro cancer-related endpoin
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1 Triphenylene (TPhen) 

2 

3 Triphenylene (CASRN 217-59-4) is an alternant PAH comprised of four fused aromatic 

4 rings. Triphenylene contains several bay regions and no fjord region in its structure. 

5 There were six datasets for triphenylene that met selection criteria and included 

6 benzo[a]pyrene (Figure 6-35); all but one of the studies gave positive results. The database 

7 includes five mutagenicity studies (four positive and one negative) and a study of in vitro DNA 

8 damage. There were no bioassays of triphenylene that met selection criteria, and no bioassays 

9 without benzo[a]pyrene. Although all of the available cancer-related endpoint studies for 

10 triphenylene gave positive results, the database is very limited, consisting of only a few in vitro 

11 mutagenicity and DNA damage studies. The RPFs for cancer-related endpoints ranged from 

12 0.02 to 0.4. Because the database for triphenylene does not provide adequate information with 

13 which to assess potential carcinogenicity, this PAH was not selected for inclusion in the RPF 

14 approach. 

15 

16 
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1 2 Figure 6-35. Triphenylene (Tphen) RPFs*. 

0.001

0.01

0.1

1

10

100

Mersch-Sundermann et 

al., 1992

Hermann, 1981 Kaden et al., 1979 Barfknecht et al., 1982 Pahlman and Pelkonen 

1987

Gibson et al., 1978

Value of RPF

Reference * Missing bar indicates nonpositive cancer-related endpoint study

Positive bioassay (incidence)Positive bioassay (multiplicity)In vivo cancer-related endpointIn vitro cancer-related endpoint
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1 7. DERIVATION OF FINAL RPFs FOR SELECTED PAHs 

2 

3 

4 The weight of evidence evaluation (Section 6) indicates that the available data are 

5 adequate to suggest that 23 of the 26 PAHs are potentially carcinogenic, three PAHs (anthracene, 

6 phenanthrene, and pyrene) exhibited little or no carcinogenic potential, and data are inadequate 

7 to evaluate the carcinogenic potential for eight PAHs. The eight PAHs with inadequate data are 

8 excluded from the RPF analysis. 

9 For the three PAHs for which there were sufficient data to conclude that the PAH had 

10 minimal carcinogenic potential (i.e., robust negative tumor bioassay data and cancer-related 

11 endpoint data), a final RPF of 0 was recommended. While there is little quantitative difference 

12 between selecting a final RPF of 0 for a given PAH and excluding that PAH from the RPF 

13 approach, this is an important distinction for uncertainty analysis. There is substantial 

14 uncertainty in the risk associated with PAHs that are excluded from the RPF analysis due to 

15 inadequate data, as these compounds could be of low or high potency. However, for PAHs with 

16 an RPF of 0, there is evidence to suggest that these compounds are of little or no carcinogenic 

17 potential, and the uncertainty associated with the cancer risk for these compounds is markedly 

18 reduced. 

19 For each of the remaining 23 compounds, a final nonzero RPF was derived. A number of 

20 options were considered for deriving a final RPF from among the numerous values calculated for 

21 each individual PAH. These options included: prioritizing bioassay RPFs from different 

22 exposure routes based on relevance to environmentally-relevant routes; prioritizing bioassay 

23 RPFs based on target organs considered relevant to human susceptibility to PAH carcinogenesis; 

24 prioritizing RPFs based on quality of the underlying study; prioritizing cancer-related endpoints 

25 by their correlation with bioassay potency (i.e., ability to predict bioassay potency); and 

26 combining RPFs across all bioassays, all cancer-related endpoints, or across all endpoints. 

27 Appendix G details analyses that were undertaken to assess various options for ranking or 

28 prioritizing RPFs. It was concluded that the available data did not provide a basis for prioritizing 

29 RPFs except for a preference for bioassay data over cancer-related endpoints. As a consequence, 

30 final RPFs were derived from bioassay data for any PAH that had at least one RPF based on a 

31 bioassay. For potentially carcinogenic PAHs without bioassay data, final RPFs were calculated 

32 from all cancer-related endpoint datasets with positive results (see next section). 

33 

34 7.1. METHODS FOR DERIVING FINAL RPFs 

35 For each potentially carcinogenic PAH with bioassay data, the average RPF was 

36 calculated from bioassay datasets with positive results (nonpositive bioassay results were not 

37 included in the calculation). For those PAHs that did not have any RPF based on a bioassay, but 

38 for which the weight of evidence evaluation indicated a potential for carcinogenic response (e.g., 
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1 dibenz[a,c]anthracene), the average RPF was calculated from all cancer-related endpoint datasets 

2 with positive results (again, nonpositive results were not included in the calculation). The range 

3 of RPF values was also reported. Presenting the average and the range provides an average and 

4 maximum estimate for each PAH that has data from multiple studies. 

5 Several options were considered for the estimation of a final RPF (e.g., arithmetic mean, 

6 geometric mean, weighted average, maximum, or order of magnitude estimates). The arithmetic 

7 mean and range were chosen as a simple approach to describing the calculated RPF values 

8 available for each PAH. Other statistical measures (i.e., geometric mean, weighted average) 

9 were not considered due to the limited number of RPF values calculated for most PAHs and the 

10 variability in the RPF estimates. There were usually not enough data (3 or fewer RPFs for 17/23 

11 PAHs with nonzero RPFs) to assess the shape of the RPF distribution for any given PAH , so a 

12 geometric mean was not considered. Further, the range of RPF values from tumor bioassays was 

13 greater than an order of magnitude for several compounds (6/23 PAHs). The variability in RPF 

14 estimates is likely due to differences in study design parameters (e.g., route, species/strain, 

15 exposure duration, exposure during sensitive time periods, initiation vs. complete carcinogenesis 

16 protocol, tumor incidence vs. tumor multiplicity reporting) and dose response methods (modeled 

17 vs. point estimates). Calculation of a weighted average was considered, but without a rationale 

18 for assigning weights among study types or tumor data outcomes, using a weighting approach 

19 might increase uncertainty. Several previous approaches for generating RPF values for PAHs 

20 have used order of magnitude estimates (Collins et al., 1998; Malcolm and Dobson, 1994; U.S. 

21 EPA, 1993; Nisbet and LaGoy, 1992, see Section 3). Providing order of magnitude estimates 

22 was not considered to be superior to calculating simple means. The presentation of the 

23 arithmetic mean and range of RPFs for each PAH was considered to be more transparent and 

24 more reflective of the available data than an order-of-magnitude approach. Including the range 

25 in the estimated RPFs was considered to be informative to the user for characterizing 

26 uncertainty. 

27 The range was reported as a measure of variability instead of a confidence interval on the 

28 average RPF. The input data for the average RPF (bioassay RPFs of different route, species, sex, 

29 and target organ, or cancer-related endpoint data across a wide variety of assays and test 

30 conditions) are likely to be correlated in unquantifiable and variable ways. There may be a high 

31 degree of correlation between RPFs calculated for a given PAH from dermal initiation and 

32 complete carcinogenicity studies, or between RPFs calculated from incidence and multiplicity 

33 data reported for the same study, but lower correlation between RPFs from dermal initiation and 

34 intraperitoneal injection. In addition, there are differences between male and female target 

35 organs after exposure to PAHs in newborn mouse tumor bioassays; RPFs from these datasets 

36 may have little or no correlation. As a result, a confidence interval on the average RPF was not 

37 calculated, but rather the range of calculated values was used as a means of expressing 

38 variability. 
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1 All tumor bioassay RPFs (across all exposure routes, species, sexes, and including both 

2 tumor incidence and tumor multiplicity RPFs) were combined to estimate the mean and range, 

3 except as follows. When separate RPFs were calculated for different endpoints in the same 

4 group of animals, the higher value of the two RPFs was included in the average and range, and 

5 the lower value was dropped from the combined data. There were two situations in which this 

6 occurred: RPFs for different target organs in the same animals, and RPFs based on incidence of 

7 tumors and tumor count in the same animals. Different RPFs were calculated for liver and lung 

8 tumors in male mice (females did not develop liver tumors) in newborn mouse studies that 

9 reported incidences or tumor counts separately. This occurrence applied only to 

10 benz[a]anthracene, chrysene, and fluoranthene tested in studies reported by LaVoie et al. (1994) 

11 and Wislocki et al. (1986). Likewise, when both incidence and multiplicity RPFs were 

12 calculated from the same experiment, the higher of the two values was included in the combined 

13 data, and the lower value was excluded. A comparison between RPFs calculated from incidence 

14 and tumor multiplicity data from the same experiment showed these values to be highly 

correlated (r215 = 0.8; see further discussion in Section 8), so RPFs from the two endpoints could 

16 not be treated as independent measures of relative potency. 

17 

18 7.2. CONFIDENCE RATINGS FOR FINAL RPFs 

19 Once a final RPF was derived for a given PAH, the resulting value was assigned a 

20 relative confidence rating of high, medium, low, or very low confidence. The relative confidence 

21 rating characterized the nature of the database upon which the final RPF was based. Confidence 

22 rankings were based on the robustness of the database. For final RPFs based on tumor bioassay 

23 data, confidence ratings considered both the available tumor bioassays and the size and 

24 consistency of the cancer-related endpoint database. The most important factors that were 

25 considered included the availability of in vivo data and whether multiple exposure routes were 

26 represented. Other database characteristics that were considered important included the strength 

27 of evidence of genotoxicity data and SAR information, the availability of more than one in vivo 

28 study, and whether effects were evident in more than one sex or species. Very low relative 

29 confidence was used to describe final RPFs based on cancer-related endpoint data only (e.g., 

30 dibenz[a,c]anthracene). 

31 For RPFs of zero, the confidence rating considered both the available tumor bioassays 

32 and the size and consistency of the cancer-related endpoint database. An RPF of zero was only 

33 applied if the data implied high or medium relative confidence. For anthracene, phenanthrene, 

34 and pyrene, it has been determined that the available data supports a practical RPF of zero. It is 

35 possible that the studies available may not provide sufficient sensitivity to compare the potency 

36 of the PAHs of interest to benzo[a]pyrene, and thus, the RPF of zero should not be considered a 

37 characterization of the inherent carcinogenicity of anthracene, phenanthrene, or pyrene. 
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1 Table 7-1 shows the average RPFs based on tumor bioassay data with their associated 

2 range and relative confidence ratings, and an overview of the tumor bioassay database (total 

3 number of studies, exposure routes tested, species tested, sexes tested, and number of RPFs 

4 derived from BMD modeling) for each PAH. Table 7-2 shows the average RPF for 

5 dibenz[a,c]anthracene, the only RPF based on cancer-related endpoint data, with its associated 

6 range, relative confidence rating, and an overview of the database for this compound. 

7 

8 
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Table 7-1. Final RPFs based on tumor bioassay data

PAH 

Average 

RPF 

Range of 

RPFs 

Relative 

confidence No. datasets Exposure routes tested 

Species 

tested 

Sexes 

tested 

RPFs ba

BMD m

Anthanthrene 0.4 0.2–0.5 Medium 2 Dermal, lung implantation Mouse, rat F 1 Anthracene 0 0 Medium 1 (Negative) Dermal Mouse F NA Benz[a]anthracene 0.2 0.02–0.4 Medium 3 Dermal, intraperitoneal Mouse F, M 0 Benz[b,c]aceanthrylene, 11H- 0.05 0.05 Low 1 Dermal Mouse F 0 Benzo[b]fluoranthene 

0.5 0.1–2 High 5 

Dermal, intraperitoneal, lung implantation Mouse, rat F, M 3 

Benz[e]aceanthrylene 0.9 0.5–1 Low 2 Dermal Mouse F, M 2 Benzo[g,h,i]perylene 0.009 0.009 Low 1 Lung implantation Rat F 1 Benz[j]aceanthrylene 60 60 Low 1 Intraperitoneal Mouse F 0 Benzo[j]fluoranthene 

0.3 0.01–1 High 5 

Dermal, intraperitoneal, lung implantation Mouse, rat F, M 2 

Benzo[k]fluoranthene 0.03 0.03–0.03 Medium 2 Dermal, lung implantation Mouse, rat F 2 Benz[l]aceanthrylene 5 4–7 Low 2 Dermal Mouse F, M 2 Chrysene 

0.1 0.04–0.2 High 7 

Dermal, intraperitoneal, lung implantation Mouse, rat F, M 3 

Cyclopenta[c,d]pyrene 0.4 0.07–1 Medium 5 Dermal, intraperitoneal Mouse F, M 2 Cyclopenta[d,e,f]chrysene, 4H- 0.3 0.2–0.5 Low 2 Dermal Mouse F 1 Dibenzo[a,e]fluoranthene 0.9 0.7–1 Low 2 Dermal Mouse F 1 Dibenzo[a,e]pyrene 0.4 0.3–0.4 Low 2 Dermal Mouse F 1 Dibenz[a,h]anthracene 

6 1–10 High 3 

Dermal, intraperitoneal, lung implantation Mouse, rat F, M 1 

Dibenzo[a,h]pyrene 0.9 0.9 Low 1 Dermal Mouse F 0 Dibenzo[a,i]pyrene 0.6 0.5–0.7 Low 2 Dermal Mouse F 1 Dibenzo[a,l]pyrene 30 10–40 Medium 3 Dermal, intraperitoneal Mouse F, M 0 Fluoranthene 0.08 0.009–0.2 Low 6 Intraperitoneal Mouse F, M 5 Indeno[1,2,3-c,d]pyrene 0.07 0.07 Low 1 Lung implantation Rat F 1 

Naphtho[2,3-e]pyrene 0.3 0.3 Low 1 Dermal Mouse F 0

Phenanthrene 

0 0 High 3 (Negative) 

Dermal, intraperitoneal, lung implantation Mouse, rat F, M NA 

Pyrene 0 0 High 7 (Negative) Dermal, intraperitoneal Mouse F, M NA 
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NA = not applicable, M = male, F = female 
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Table 7-2. Final RPFs based on cancer-related endpoint data 

(no tumor bioassay data available) 

PAH 

Average 

RPF 

Range of 

RPFs 

Relative 

confidence Types of studies Multiple dose studies

Dibenz[a,c]anthracene 4 0.04–50 Very low Total = 14 studies

One in vivo DNA adduct 

Six in vitro bacterial 

mutagenicity 

One in vitro mammalian 

mutagenicity 

One in vitro 

morphological/malignant 

transformation 

Three in vitro DNA 

damage 

Two in vitro DNA adducts

Total = 6 studies

Four in vitro bacterial 

mutagenicity 

One in vitro DNA 

damage 

One in vitro DNA 

adduct 

1 

2 
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1 7.3. SUSCEPTIBILITY FROM EARLY LIFE EXPOSURE TO CARCINOGENS 

2 According to the Supplemental Guidance for Assessing Susceptibility from Early Life 

3 Exposure to Carcinogens (U.S. EPA, 2005b), benzo[a]pyrene is carcinogenic by a mutagenic 

4 mode of action. For example, an acute dosing study using benzo[a]pyrene suggests that early5 lifestage exposure would lead to an increased incidence of tumors compared with adult 

6 exposures of a similar dose and duration (EPA 2005b). Mice that were treated with 

7 benzo[a]pyrene (75 or 150 μg/g body weight intraperitoneal) within 24 hours of birth or at 15 

8 days of age developed hepatomas at a higher incidence than similarly treated animals at 42 days 

9 of age (Vesselinovitch et al., 1975, as cited in EPA 2005b. 

10 The Supplemental Guidance for Assessing Susceptibility from Early-Life Exposure to 

11 Carcinogens establishes ADAFs for three specific age groups. The current ADAFs and their age 

12 groupings are 10 for <2 years, 3 for 2 to <16 years, and 1 for 16 years and above (U.S. EPA, 

13 2005b). The 10-fold and 3-fold adjustments in slope factor are to be combined with age specific 

14 exposure estimates when estimating cancer risks from early life (<16 years age) exposure to 

15 PAHs. 

16 Because a mutagenic mode of action for benzo[a]pyrene carcinogenicity is sufficiently 

17 supported in laboratory animals and relevant to humans, and in the absence of chemical-specific 

18 data to evaluate differences in susceptibility, increased early-life susceptibility is assumed and 

19 the age-dependent adjustment factors (ADAFs) should be applied, as appropriate. 

20 The 23 PAH compounds for which a RPF value was determined (see Table 7-2) are also 

21 considered to be carcinogenic by a mutagenic mode of action (see Section 2.4 for discussion of 

22 similarities in mode of action for PAHs). In the absence of chemical-specific data to evaluate 

23 differences in susceptibility, increased early-life susceptibility to the 23 PAHs (for which RPFs 

24 were derived) in this analysis is assumed and the age-dependent adjustment factors (ADAFs) 

25 should be applied, along with exposure information, as appropriate. When assessing PAH cancer 

26 risks, the RPF values should be applied to the benzo[a]pyrene risk estimates with adjustment for 

27 early life susceptibility (See Table 7-3 for example). 

28 

Table 7-3. Sample calculation of estimated cancer risk for 

benz[a]anthracene (BaA) with the application of ADAFs 

age 

group 

ADAF 

B[a]P oral slope 

factor (per 

mg/kg-day) 

adjusted 

B[a]P 

cancer risk 

estimate 

RPF 

BaA estimated 

cancer risk (per 

mg/kg-day) 

0 < 2 10 7.3 73 0.2 15 

2<16 3 7.3 24 0.2 4.8 

16+ 1 7.3 7.3 0.2 1.5 

29 
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1 

2 8. UNCERTAINTIES ASSOCIATED WITH RPF APPROACHES 

3 

4 

5 A description of uncertainties and limitations is an important component of the RPF 

6 approach for PAH mixtures risk assessment. Many of the general uncertainties related to 

7 chemical-specific risk assessment are also applicable to the proposed RPF approach for PAHs. 

8 These include issues related to selection of an appropriate animal model, low-dose and 

9 interspecies extrapolation, and variability within the human population. Use of a component10 based approach to mixtures risk assessment leads to additional uncertainties, e.g., the lack of 

11 experimental data on potential interactions among individual components within the mixture 

12 (i.e., among PAHs and with other chemicals). 

13 The feasibility of conducting a robust component-based approach for PAH mixtures 

14 (RPF approach) was evaluated by a PAH mixtures peer consultation workshop (U.S. EPA, 

15 2002). Included in the discussion was a general evaluation of U.S. EPA’s Provisional Guidance

16 (U.S. EPA, 1993). Workshop participants highlighted the following limitations of the 1993 

17 guidance: 

18 

19 (1) The approach only considered a small subset of PAHs (that is unsubstituted PAHs only, 

20 no heterocyclic compounds or nitro- or alkyl-substituted PAHs); 

21 

22 (2) There are no human toxicity data for any individual PAH; 

23 

24 (3) The assumption of additivity may not be valid, and there may be interactions among 

25 PAHs or between PAHs and other components of a mixture (e.g., metals); 

26 

27 (4) PAHs may generally have a common mode of action (i.e., mutagenicity), but multiple 

28 modes of action for carcinogenesis are possible; and 

29 

30 (5) The estimated order of potency (EOPP) approach was limited to the oral exposure route 

31 (i.e., a recommendation was made not to apply the factors to dermal and inhalation 

32 exposures). 

33 

34 The current analysis represents a significant improvement upon the previous component35 based approach to PAH mixtures risk assessment. One of the most important improvements is a 

36 comprehensive review of the scientific literature dating from the 1950s through 2009 on the 

37 carcinogenicity and genotoxicity of PAHs. The search identified over 900 individual 

38 publications for a target list of 74 PAHs that had been identified in environmental media or for 

39 which toxicological data were available. Review of these publications resulted in the 

40 identification of more than 600 papers that included carcinogenicity or cancer-related endpoint 

41 data on at least one PAH and benzo[a]pyrene tested at the same time. Dose-response data were 

42 extracted, and individual RPFs were calculated from over 300 data sets representing 50 
198 DRAFT – DO NOT CITE OR QUOTE 

1 individual PAHs. A weight of evidence evaluation was conducted to evaluate the evidence for 

2 potential carcinogenicity of 34 of these PAHs; data were inadequate to conduct such an 

3 evaluation for the remaining 16 compounds. A final RPF was derived for each PAH based on 

4 tumor bioassay data (if available) or cancer-related endpoint data if no tumor bioassay RPFs 

5 were available. Final RPFs were derived for 26 PAHs (see Table 7-2 in Section 7), significantly 

6 increasing the number of PAHs that can be addressed through this approach. Each RPF was 

7 assigned a relative confidence rating reflecting the size and diversity of the tumor bioassay or 

8 cancer-related endpoint database that was used to derive the final RPF for that PAH. 

9 Despite these improvements, many of the uncertainties highlighted during the 2002 peer 

10 consultation workshop (U.S. EPA, 2002) also apply to the current analysis. The following 

11 sections describe some specific uncertainties and limitations associated with the development 

12 and use of RPFs for PAHs. The uncertainties that are specific to the approach presented herein 

13 are discussed below in Sections 8.1 and 8.2. The remaining sections (8.3–8.6) discuss the 

14 general uncertainties associated with a component-based approach to PAH mixtures risk 

15 assessment. These include the number of PAHs included in the approach, human relevance of 

16 animal data, assumptions regarding mode of action and dose additivity, and cross-route 

17 extrapolation. 

18 

19 8.1. UNCERTAINTY IN DOSE-RESPONSE FOR INDIVIDUAL PAHS 

20 Several uncertainties and limitations are specifically associated with the dose-response 

21 assessment methodology used in this analysis to derive RPFs for PAHs. Uncertainties are 

22 associated with the following decisions: 

23 

24 • Use of a single dose-response model for quantal or continuous data; 

25 

26 • Use of varying BMR levels; 

27 

28 • Use of tumor incidence data at the upper end of the dose-response curve (e.g., greater that 

29 75% incidence) to calculate some RPFs; 

30 

31 • Use of tumor multiplicity data to calculate some RPFs; 

32 

33 • Use of single-dose point estimates to calculate some RPFs; 

34 

35 • Reliance on data from cancer-related endpoint studies in the absence of bioassays; and 

36 

37 • Use of cancer-related data from assay conditions that maximize the benzo[a]pyrene 

38 response, even though these conditions were not necessarily optimal for other PAHs. 

39 

40 The decision was made to employ a single dose-response model for either quantal or 

41 continuous data due to the large number of data sets that needed be analyzed from the PAH 
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1 database. The multistage model for incidence data and the linear model for continuous data were 

2 considered to be broadly applicable to different types of data as simple curve-fitting models. In 

3 some cases, the goodness-of-fit criteria indicated that the selected model did not fit the data. In 

4 these cases, high-dose groups were sequentially eliminated until an adequate fit was achieved, 

5 but other model structures (e.g., gamma, probit, logistic, etc.) were not considered. 

6 Tumor bioassay data were modeled at a BMR of 10% (extra risk above control) in order 

7 to target the low end of the dose-response curve as the point of departure for slope estimation. 

8 When this was not feasible, usually because only a single dose was used for benzo[a]pyrene, an 

9 attempt was made to match individual target PAH response levels to the benzo[a]pyrene 

10 response chosen for the point estimate. This assumes that the shape of the dose-response curve 

11 is similar for the target PAH and benzo[a]pyrene (also a necessary assumption of dose additivity) 

12 and that the slope is constant across the dose-response curve. These assumptions may not hold, 

13 especially in studies of tumor incidence where the point estimate benzo[a]pyrene response was 

14 very high or near maximal. In many cases, the dose of benzo[a]pyrene selected as the positive 

15 control produced near maximal tumor incidence in exposed animals (i.e., >75%). There is 

16 uncertainty associated with comparing potency estimates at the high end of the dose-response 

17 curves and using the resultant RPF to estimate risks associated with low environmental 

18 exposures. The relative potency relationship between any two PAHs may be different at the low 

19 end, compared with the high end, of the dose-response curves. 

20 It is not clear whether relative potency values estimated at the high end of the dose21 response curve are reasonably predictive of relative potency at low environmental exposure 

22 levels. For this reason, additional uncertainty is involved in using RPFs that are not based on a 

23 BMR of 10% (especially those RPFs that are based on responses exceeding 75%) to estimate 

24 risks associated with low exposures. 

25 If model fit was not achieved, then a point-estimate ratio approach was used. Point 

26 estimate ratios were also used for several other reasons: 

27 

28 (1) Only a single dose group was tested; 

29 

30 (2) When the standard deviation or number of replicates were not reported for continuous 

31 data sets; or 

32 

33 (3) High-dose groups from multiple dose data sets were not usable due to a saturated tumor 

34 response (>90% incidence in the lowest exposure group). 

35 

36 The point estimate approach is most reliable when the chosen point is in the linear 

37 portion of the dose-response curve. In many cases, however, especially for single-dose data, it 

38 was not possible to determine whether the chosen point was in a linear or nonlinear portion of 

39 the dose-response curve. The dose-response relationship observed in many studies of cancer-
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1 related endpoints was nonlinear at high doses. Whenever possible, the point estimate was chosen 

2 from the linear portion of the dose-response curve (i.e., before the response plateau that occurs at 

3 high doses). Of 50 individual RPFs calculated from tumor incidence data, 19 were calculated 

4 using a point of departure incidence ≤25%, 21 were calculated using a point of departure 

5 incidence between 25 and 75%, and the remaining 10 were calculated using a point of departure 

6 incidence between 75 and 90%. Thus, only 20% of the individual RPFs for tumor incidence data 

7 were calculated from a point high (>75 and <90% incidence) on the dose-response curve. 

8 For a few PAHs tested in older dermal bioassays , the authors reported mortality prior to 

9 the appearance of the first tumor. For these data sets, an assumption was made that the number 

10 of animals at risk for tumor development was equal to the total number of animals alive at the 

11 time of the appearance of the first tumor. This approach ensures that the incidence is not 

12 underestimated by including animals that did not survive long enough to develop tumors. As this 

13 assumption applied to a small number of RPFs (specifically, individual RPFs for chrysene, 

14 dibenzo[a,e]pyrene, dibenzo[a,e]fluoranthene, and dibenzo[a,h]pyrene calculated from data 

15 reported by Hecht et al. [1974] and Hoffmann and Wynder [1966]), it had little impact on the 

16 overall analysis. However, as the final RPFs for dibenzo[a,e]pyrene, dibenzo[a,e]fluoranthene, 

17 and dibenzo[a,h]pyrene are based exclusively on the data reported by Hoffmann and Wynder 

18 (1966), there is additional uncertainty in these values stemming from the occurrence of early 

19 mortality. 

20 RPFs were calculated for many cancer-related endpoints. Many of the studies describing 

21 in vitro cancer-related endpoints provided dose-response data under varying study conditions. 

22 For example, bacterial mutagenesis studies utilized multiple strains, different metabolic 

23 activation processes, and varying assay systems. In order to minimize the amount of data used 

24 for dose-response analysis of in vitro mutagenicity studies, and to provide a consistent basis for 

25 comparing RPFs for differenct PAHs, the data from conditions that maximize the 

26 benzo[a]pyrene response within a particular study were used for the dose-response assessment. 

27 In several studies, the conditions that were optimal for benzo[a]pyrene were not necessarily 

28 optimal for the target PAH. For example, the concentration of S9 mix that produced the highest 

29 mutation rate for benzo[a]pyrene did not produce a maximal response for perylene or 

30 cyclopenta[c,d]pyrene (Carver et al., 1986; Eisenstadt and Gold, 1978). In vitro data were only 

31 used in the derivation of a single final RPF (for dibenz[a,c]anthracene; see Table 7-2); thus, the 

32 uncertainties associated with the use of cancer-related endpoint data are important for 

33 dibenz[a,c]anthracene but have minimal impact on the proposed RPFs for the other 25 PAHs. 

34 

35 8.2. UNCERTAINTY IN SELECTING PAHs FOR INCLUSION IN RPF APPROACH 

36 One of the uncertainties highlighted by the peer consultation workshop (U.S. EPA, 2002) 

37 stemmed from the fact that U.S. EPA’s 1993 provisional EOPP approach only considered a small 

38 subset of PAHs (i.e., unsubstituted PAHs only, no heterocyclic compounds or nitro- or alkyl-
201 DRAFT – DO NOT CITE OR QUOTE 

1 substituted PAHs), and EOPPs were available for only seven PAHs. Although the present report 

2 considered a larger number of PAHs than previous analyses (the toxicological literature was 

3 searched for data on 74 individual PAHs identified in environmental media or for which there 

4 were toxicological data), the focus of this analysis remains limited to unsubstituted PAHs with 

5 three or more fused aromatic rings containing only carbon and hydrogen atoms. Thus, the RPF 

6 analysis presented here does not account for the possible carcinogenicity of substituted or 

7 heterocyclic PAHs that may be present in complex mixtures. This may result in an 

8 underestimation of PAH mixture cancer risk. 

9 Of the 74 unsubstituted PAHs with three or more aromatic rings, there were studies 

10 including benzo[a]pyrene that were suitable for RPF calculation for 50 compounds. The 

11 methodology for selecting PAHs for inclusion in the RPF approach from among these 50 PAHs 

12 is described in Section 6. At the outset, 16 PAHs were excluded because only one or two in vitro 

13 cancer-related endpoint RPFs were available. The remaining 34 were evaluated using a weight 

14 of evidence approach. The primary uncertainties associated with the selection process relate to: 

15 

16 (1) The use of a weight of evidence approach that focused on tumor bioassays including 

17 benzo[a]pyrene as opposed to a comprehensive cancer assessment to select PAHs for 

18 inclusion in the approach; and 

19 

20 (2) The exclusion of PAHs with limited or inconclusive data. 

21 

22 The weight of evidence approach was used due to the large number of compounds that 

23 were under consideration. The approach was structured as a decision tree that focused primarily 

24 on cancer bioassays that included benzo[a]pyrene, and only considered other data (e.g., bioassays 

25 that did not include benzo[a]pyrene, or cancer-related data) when cancer bioassays with 

26 benzo[a]pyrene were unavailable, nonpositive, or inconsistent (see Figure 6-1). The data 

27 collection for this analysis was centered on studies that included benzo[a]pyrene, as these studies 

28 would be most useful for RPF calculation. Consequently, information from bioassays that 

29 included benzo[a]pyrene were readily available for use in the weight of evidence determinations. 

30 Bioassays that did not include benzo[a]pyrene and cancer-related endpoint data were considered 

31 only when there were conflicting or negative results in the studies that did include 

32 benzo[a]pyrene. There is uncertainty in drawing conclusions as to potential carcinogenicity 

33 based on a narrow subset of the available database. Other elements of a more comprehensive 

34 weight of evidence determination that were not considered include: cancer-related endpoint data 

35 from studies that did not include benzo[a]pyrene; information on tumorigenicity of metabolites; 

36 information on formation of reactive metabolites; other mechanistic data (e.g., Ah reactivity, 

37 inhibition of gap junction intercellular communication, etc.); and quantitative structure-activity 

38 assessment. 
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1 A number of PAHs (24 of 50 PAHs that had at least one RPF value) were excluded from 

2 the relative potency approach because the available data were inadequate to draw a conclusion as 

3 to potential carcinogenicity (see Tables 6-1 and 6-2). All of these PAHs had at least one RPF, 

4 indicating that the compounds were active in at least one cancer-related endpoint assay. 

5 Excluding these PAHs from the approach increases the uncertainty in assessing risks from a 

6 mixture that includes them, particularly if the excluded PAHs constitute a large fraction of the 

7 mixture. 

8 In final, RPFs were proposed for only 26 of the 74 PAHs initially considered, because the 

9 remaining 48 compounds did not have adequate data. Thus, even among the subset of PAHs 

10 upon which this analysis was focused, RPFs were only recommended for only about one-third of 

11 the compounds. Because only a fraction of any given PAH mixture can be evaluated using the 

12 RPF approach, it will be important to provide an evaluation of the proportion of the total mixture 

13 (i.e., mass fraction) that is comprised of compounds that are not considered in the component14 based approach as part of the uncertainty evaluation of a risk assessment using these RPFs. 

15 

16 8.3. UNCERTAINTY IN DERIVING A FINAL RPF FOR EACH PAH 

17 The methodology for deriving a final RPF value and assigning a relative confidence 

18 rating is described in Section 6.1. The primary uncertainties associated with RPF derivation 

19 relate to: 

20 

21 (1) Combining RPFs across multiple exposure routes, species, sexes, tumor types, and 

22 studies; 

23 

24 (2) Inclusion of RPFs based on tumor multiplicity data in the combined data; 

25 

26 (3) Use of an arithmetic mean to derive final RPFs; and

27 

28 (4) Use of cancer-related endpoint data to derive final RPFs for compounds without tumor 

29 bioassay RPFs. 

30 

31 A variety of options were considered for prioritizing and/or combining RPFs. Appendix 

32 G describes analyses that were undertaken to assess options for prioritizing RPFs. As the 

33 appendix indicates, the current state of knowledge does not suggest a clear biological basis for 

34 prioritizing RPFs. As a result, RPFs were combined across exposure routes, species, sexes, 

35 tumor types, dose-response methods, and studies. 

36 In addition to tumor incidence data, tumor multiplicity data were used to calculate RPFs. 

37 In some instances, tumor incidence data could not be used for RPF derivation (e.g., the incidence 

38 at the lowest dose was in the plateau region of the dose-response curve; ≥90% incidence), while 

39 tumor multiplicity data were available. The relationship between tumor incidence RPFs and 

40 tumor multiplicity RPFs is not known; however, this analysis resulted in the calculation of both 
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1 incidence and multiplicity RPFs for a number of studies. These data were plotted, and a linear 

2 regression analysis was performed to assess the correlation between these two relative potency 

3 estimates. Figure 8-1 shows the results. 

4 

0.01

0.1

1

10

0.01 0.1 1 10

Incidence RPF

Multiplicity RPF

1:1 Correspondence Line

5 

6 

7 Figure 8-1. Correlation between incidence and multiplicity RPFs. 

8 

9 As shown in Figure 8-1, there is a high degree of correlation between incidence and 

10 multiplicity RPFs calculated from results in the same animals. The regression analysis indicated 

an r

2

11 of 0.80 for the correlation. The figure also shows that multiplicity RPFs exhibit a slight 

12 tendency to underestimate the RPF from incidence data (more points are to the right of the 

13 1:1 correspondence line). Nevertheless, the correlation plot and regression results provide 

14 support for the use of RPFs from tumor multiplicity data when incidence data were not available 

15 or not useable. 

16 As the incidence and multiplicity RPFs from the same study were highly correlated, only 

17 one of the two metrics was included in the combined RPFs. Specifically, the higher of the 

18 incidence or multiplicity RPF from the same study was included in the average and range. 

19 Consistent with the figure, the higher value was usually calculated from incidence data. 

20 Final RPFs were calculated as the arithmetic mean and range of RPFs from tumor 

21 bioassay data when such data were available. Presenting the average and the range provides both 

22 an average and a maximum estimate for each PAH that has data from multiple studies. Other 
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1 options for deriving a central tendency RPF include geometric mean, median, weighted average, 

2 and order of magnitude estimates. The arithmetic mean represents a simple approach to 

3 describing the calculated RPF values available for each PAH. There were usually not enough 

4 data (≤3 RPFs for 17/23 PAHs with nonzero RPFs) to assess the shape of the RPF distribution 

5 for any given PAH, so a geometric mean was not considered. Calculation of a weighted average 

6 was considered, but without a clear biological rationale for assigning weights among study types 

7 or tumor data outcomes, using a weighting approach might increase uncertainty. Finally, 

8 providing order of magnitude estimates, as has been previously done for estimating RPFs for 

9 PAHs, was not considered to be superior to calculating simple means. Including the range in the 

10 estimated RPFs was considered to be informative to the user for characterizing uncertainty. 

11 Cancer-related endpoint data were relied upon for the derivation of a RPF for only one 

12 PAH (dibenz[a,c]anthracene). For this compound, there were no tumor bioassay data suitable for 

13 the determination of an RPF. However, cancer-related endpoint data provided qualitative 

14 support for the finding of potential carcinogenicity for this compound (see individual narrative 

15 for this compound in Section 6.2). Although the mode of action for PAHs suggests that, in 

16 general, these endpoints may be relevant to PAH carcinogenicity, the predictive value of a 

17 positive response in these tests has not been conclusively demonstrated. Thus, there is 

18 considerable uncertainty in an RPF based on cancer-related endpoint data. Appendix G includes 

19 analysis of the correlation between average RPFs calculated from cancer-related endpoint data 

20 and tumor bioassay data. As shown in Table 8-1, and further discussed in Appendix G, cancer21 related endpoint RPFs are reasonably predictive of tumor bioassay RPFs; however, the 

22 relationship between these RPFs and the relative potency of a given PAH in humans exposed via 

23 environmentally-relevant routes is unknown. 

24 

Table 8-1. Results of simple linear regression of log–transformed average 

genotoxicity RPF vs. log average tumor bioassay RPF

Genotoxicity endpoint R

2

 Slope p-Value n 

All in vivo DNA adducts 0.64 1.24 <0.01 9 

All in vivo non-bioassays 0.54 1.05 0.016 10 

All non-bioassay endpoints (in vitro and in vivo) 0.43 1.03 <0.01 19 

All in vitro non-bioassays 0.39 0.91 <0.01 19 

All in vivo micronuclei and sister chromatid exchanges 0.58 0.83 >0.05 (NS) 6 

All in vitro mutagenicity 0.047 0.39 >0.05 (NS) 17 

25 

26 For three PAHs (anthracene, phenanthrene, and pyrene), a final RPF of 0 was 

27 recommended. As noted earlier in Section 6, there is little quantitative difference between 

28 selecting a final RPF of 0 for a given PAH and excluding that PAH from the RPF approach. 

29 However, excluding PAHs from the RPF approach implies substantial uncertainty (these 

30 compounds could be of low or high potency), while assigning an RPF of 0 suggests lower 
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1 uncertainty because there is evidence to suggest that these compounds are of little or no 

2 carcinogenic potential. Nevertheless, there remains uncertainty in the RPFs for these three 

3 compounds, as all of them included one or more studies suggesting activity in cancer-related 

4 endpoint assays. 

5 In the present analysis, RPFs for individual PAHs were based on data of varying quality 

6 and reproducibility, so there is additional uncertainty in risks estimated for mixtures containing 

7 differing concentrations of individual PAHs. Confidence ratings were assigned to each RPF to 

8 qualitatively characterize the uncertainty in each individual RPF. Table 8-2 shows the 

9 distribution of PAHs with RPFs of each confidence rating. As the table indicates, there are 

10 6 PAHs with RPFs of high confidence, 6 PAHs with RPFs of medium confidence, 14 PAHs with 

11 RPFs of low confidence, and 1 PAH with an RPF of very low confidence. The confidence 

12 ratings assigned to the RPFs may be used to qualitatively assess the uncertainty in a mixtures 

13 risk assessment that utilizes the RPFs. For example, if a high proportion of the total cancer risk 

14 predicted for a given mixture is attributable to benzo[a]pyrene and other PAHs with RPFs of 

15 high or medium confidence, then the confidence in the overall cancer risk assessment will be 

16 relatively high. If, in contrast, benzo[a]pyrene contributes a relatively small fraction of the 

17 overall risk, and/or the mixture consists primarily of PAHs with RPFs of low confidence, then 

18 the confidence in the overall cancer risk assessment will be correspondingly lower. Thus, it will 

19 be important to consider the relative contribution of benzo[a]pyrene to the total risk, as well as 

20 the relative confidence ratings of the RPF values for component PAHs, in the uncertainty 

21 evaluation for cancer risk assessments that employ these RPFs. 

22 

Table 8-2. PAHs with RPFs of varying confidence 

High confidence RPF Medium confidence RPF Low confidence RPF 

Very low confidence 

RPF 

Benzo[b]fluoranthene Anthanthrene Benz[b,c]aceanthrylene, 11H- Dibenz[a,c]anthracene 

Benzo[j]fluoranthene Anthracene Benz[e]aceanthrylene 

Chrysene Benz[a]anthracene Benzo[g,h,i]perylene 

Dibenz[a,h]anthracene Benzo[k]fluoranthene Benz[j]aceanthrylene 

Phenanthrene Cyclopenta[c,d]pyrene Benz[l]aceanthrylene 

Pyrene Dibenzo[a,l]pyrene Cyclopenta[d,e,f]chrysene, 4H-

 Dibenzo[a,e]fluoranthene 

 Dibenzo[a,e]pyrene 

 Dibenzo[a,h]pyrene 

 Dibenzo[a,i]pyrene 

 Fluoranthene 

 Indeno[1,2,3-c,d]pyrene 

 Naphtho[2,3-e]pyrene 

23 
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1 8.4. UNCERTAINTY IN USE OF ANIMAL DATA TO PREDICT HUMAN CANCER 

2 RISK 

3 Section 4.2 briefly summarizes the epidemiology and human biomarker data related to 

4 exposure to PAH mixtures and carcinogenicity. Exposure to certain PAH mixtures is clearly 

5 associated with cancer in humans. Epidemiology studies evaluating emissions from coke 

6 production, coal gasification, aluminum production, iron and steel founding, coal tars, coal tar 

7 pitches, and soot have demonstrated associations between exposure and increased risk of lung 

8 cancer in humans (see review of Bostrom et al., 2002). Skin and scrotal cancers have been 

9 associated with exposure to coal tar, coal tar pitches, non-refined mineral oils, shale oils, and 

10 soot (Larsen and Larsen, 1998; WHO, 1998; ATSDR, 1995). While human epidemiology data 

11 may be sufficient for the purpose of quantifying the cancer risks associated with exposure to a 

12 few PAH mixtures, there are no data for many mixtures; hence the need for other approaches 

13 including surrogate-mixture and component-based approaches. As noted by the peer 

14 consultation workshop (U.S. EPA, 2002), there are no human data on cancer response to 

15 individual PAHs that could be used as the basis for, or as a supplement to, a component-based 

16 approach. As a result, the RPF approach relies on animal bioassay data to predict human cancer 

17 risk associated with individual PAHs. 

18 The use of animal bioassays in predicting relative carcinogenic potency in humans 

19 represents a source of uncertainty in this approach. As there are no human data on cancer 

20 response to individual PAHs, including benzo[a]pyrene, there can be no quantitative evaluation 

21 of uncertainty in extrapolating from RPFs based on animal bioassay data to relative potency in 

22 humans. Possible species differences in toxicokinetics, toxicodynamics, and mode of action 

23 contribute to the uncertainty. Cancer-related endpoint data are available using human cells (e.g., 

24 epidermal keratinocytes, lymphoblasts, human epithelial cells) for the evaluation of 

25 mutagenicity, DNA adducts, unscheduled DNA synthesis, DNA damage, and clastogenicity or 

26 sister chromatid exchange frequency (see Section 4.3). Findings in human cells were generally 

27 consistent with those in other mammalian cells; however, whether this finding of consistency 

28 extends to effects in vivo, and specifically to formation of tumors, is not known. 

29 In addition, animal bioassays use various routes of administration (e.g., intraperitoneal 

30 and subcutaneous injection), which may not be directly relevant to expected routes of exposure 

31 for humans. It is difficult to determine whether the relative potency based on animal bioassays 

32 using injection routes of exposure is predictive of relative potency that would be observed in 

33 humans exposed through environmentally relevant exposure routes (see further discussion of 

34 exposure-route uncertainties in Section 6.6). An additional source of uncertainty in the use of 

35 animal bioassay data stems from differences in the doses used in animal bioassays as compared 

36 with low doses received by humans exposed in the environment. Further discussion of this issue 

37 as it relates to dose-response modeling is provided in Section 6.1. 
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1 Mechanistic data, primarily obtained using benzo[a]pyrene, provide support for the 

2 human relevance of PAH tumorigenicity in animals. There is evidence linking three pathways 

3 activating benzo[a]pyrene to DNA-reactive agents [(+)-anti-BPDE, radical cations, 

4 benzo[a]pyrene-7,8-dione, and reactive oxygen species] with key mutational events in genes 

5 (p53 tumor suppressor gene and H-ras or K-ras oncogenes) that can lead to tumor initiation. 

6 Results in support of mutagenic modes of action via the diol epoxide and radical cation pathways 

7 include in vivo results in animals. All of these activation pathways occur in human tissues, and 

8 associations have been made between spectra of mutations in the p53 tumor suppressor gene or 

9 ras oncogenes induced by benzo[a]pyrene metabolites with spectra of mutations in these genes in 

10 tumor tissue from benzo[a]pyrene-exposed animals or tumor tissue in humans. 

11 Support for the association between the diol epoxide pathway and tumor initiation 

12 includes observation that: (+)-anti-BPDE activated the H-ras-1 proto-oncogene to transform 

NIH/3T3 cells via G→T point mutations in the 12th 13 codon (Marshall et al., 1984); (+)-anti14 BPDE reacts with the p53 tumor suppressor gene at several hotspots mutated in lung cancer 

15 patients (Denissenko et al., 1996; Puisieux et al., 1991); the spectra of p53 and K-ras mutations 

16 in lung tumors of nonsmoking patients, chronically exposed to smoky coal emissions, was 

17 consistent with (+)-anti-BPDE mutations in these genes (DeMarini et al., 2001); elevated BPDE18 DNA adducts have been observed in coke oven workers and chimney sweepers (Pavanello et al., 

19 1999); and the spectra of mutation in the K-ras, H-ras, and p53 genes in forestomach tumors of 

20 mice fed benzo[a]pyrene in the diet for 2 years were consistent with (+)-anti-BPDE DNA 

21 reactions (Culp et al., 2000). 

22 Support for the radical cation pathway includes observations that depurinated adducts, 

23 (expected products from reactions of benzo[a]pyrene radical cations with DNA) accounted for 

24 74% of identified DNA adducts in mouse skin exposed to benzo[a]pyrene (Rogan et al., 1993) 

25 and 9/13 examined tumors from mice exposed to dermal applications of benzo[a]pyrene had H26 ras oncogene mutations attributed to depurinated DNA adducts from benzo[a]pyrene radical 

27 cations (Chakravarti et al., 1995). 

28 Support for the AKR pathway includes in vitro demonstration that several types of DNA 

29 damage can occur from o-quinones and reactive oxygen species (Park et al., 2006; Balu et al., 

30 2004; McCoull et al., 1999; Flowers-Geary et al., 1997, 1996), benzo[a]pyrene-7,8-dione can 

31 induce mutations in the p53 tumor suppressor gene using an in vitro yeast reporter gene assay 

32 (Park et al., 2008; Shen et al., 2006; Yu et al., 2002), and dominant p53 mutations induced by 

33 benzo[a]pyrene,7,8-dione in this system corresponded with p53 mutation hotspots observed in 

34 human lung cancer tissue (Park, 2008). 

35 All three activation pathways are expected to occur in human tissues (Jiang et al., 2007), 

36 and associations have been made between spectra of mutations in the p53 tumor suppressor gene 

37 or ras oncogenes induced by benzo[a]pyrene metabolites with spectra of mutations in these genes 

38 in tumor tissue from benzo[a]pyrene-exposed animals or humans. In particular, DeMarini et al. 
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1 (2001) demonstrated mutations in the p53 tumor suppressor gene and the K-ras oncogene in the 

2 lung tumors of nonsmokers, whose tumors were associated with exposure to smoky coal. 

3 The available information supporting these actions for benzo[a]pyrene is consistent with 

4 what is known about the mode of action for other PAHs demonstrated to induce cancer in 

5 animals, including cyclopenta[cd]pyrene, dibenz[a,h]anthracene, and dibenzo[a,l]pyrene 

6 (Cogliano et al., 2008; Straif et al., 2005). All PAHs that have been studies require metabolic 

7 activation to produce carcinogenic responses in animals and there is evidence for activation to 

8 DNA reactive intermediates via several pathways (Straif et al., 2005; Xue and Warshawsky, 

9 2005; WHO, 1998; Cavalieri and Rogan, 1995). For example, incubation of rat liver 

10 microsomes with dibenzo[a,l]pyrene, a PAH that is more tumorigenically potent than 

11 benzo[a]pyrene in mouse skin and rat mammary tissue, formed depurinated DNA adducts from 

12 the radical cation pathway, as well as DNA adducts from the diol epoxide pathway (Cavalieri 

13 and Rogan, 1995). 

14 In summary, the relevance of animal bioassay data to the prediction of human 

15 carcinogenic potency remains a significant area of uncertainty in the use of this and other 

16 approaches to PAH cancer risk assessment. However, mechanistic data on benzo[a]pyrene and 

17 other PAHs provide evidence that the molecular events leading to PAH-induced tumor formation 

18 in animals are relevant to humans. 

19 

20 8.5. UNCERTAINTY IN THE ASSUMPTIONS OF COMMON MODE OF ACTION 

21 AND DOSE ADDITIVITY 

22 A discussion of the potential modes of action for PAH carcinogenicity is presented in 

23 Section 2.4. Individual carcinogenic PAHs are linked by a common effect (i.e., tumorigenicity), 

24 which may occur through multiple mechanisms. Reactive metabolites produced during 

25 metabolic transformations of PAHs include diol epoxides, reactive oxygen species, radical 

26 cations, and o-quinones. The formation of these metabolites is not mutually exclusive, and the 

27 carcinogenic process for PAHs is likely to be related to some combination of molecular events 

28 resulting from formation of several reactive species. Reactive metabolites of PAHs interact with 

29 DNA to form adducts and produce DNA damage resulting in mutations in cancer-related genes 

30 such as tumor suppressor genes or oncogenes. These events appear to reflect the initiation 

31 potency of an individual PAH (e.g., strong mutagens are generally potent initiators) (Sjogren et 

32 al., 1996). Certain PAHs exhibit promotional effects that may be related to cytotoxicity and the 

33 formation of reactive oxygen species, AHR affinity and the upregulation of genes related to 

34 biotransformation (i.e., induction of CYP1A1), growth, and differentiation (Bostrom et al., 

35 2002). The inhibition of gap junctional intracellular communication is also related to tumor 

36 promotion by PAHs (Bostrom et al., 2002). The ability of certain PAHs to act as tumor 

37 promoters as well as initiators may increase their carcinogenic potency in animal bioassays 

38 conducted at high doses. Initiation potency may be more relevant to low level environmental 
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1 exposure in humans (Bostrom et al., 2002; Sjogren et al., 1996); however, the proposed RPF 

2 approach is not unduly affected by this as it relies largely on high dose animal bioassay data for 

3 selecting RPF values. This represents an uncertainty in the use of the RPF approach in 

4 estimating human cancer risks from PAHs. 

5 Conceptually, the uncertainty related to relative potency for initiation versus promotion 

6 could be reduced by using separate RPF schemes for each part of the carcinogenic process. This 

7 would require selection of indicator compounds that best represent the initiation and promotion 

8 processes, and use of mechanistic data to determine relative potency for each process (i.e., 

9 mutagenicity for initiation, AhR binding or enzyme induction for promotion). There are several 

10 problems with this approach, including the lack of data to support the selection of indicator 

11 compounds and the complete carcinogenic nature of many PAHs (i.e., they act as both initiators 

12 and promoters). The initiation and promotion potency of an individual PAH is determined by its 

13 chemical structure. Some PAHs are strong mutagens, but have low affinity for the AHR (e.g., 

14 fjord region PAHs) (Bostrum et al., 2002; Sjogren et al., 1996). Other PAHs are complete 

15 carcinogens, with initiating properties (i.e., mutagenesis) and AhR affinity leading to tumor 

16 promotion (e.g., benzo[a]pyrene, dibenz[a,h]anthracene) (Bostrum et al., 2002; Sjogren et al., 

17 1996). Benzo[a]pyrene is considered a good indicator compound for similar PAHs with 

18 complete carcinogenic activity. However, the relative potency of other PAHs, especially those 

19 that act primarily via either initiation or promotion, may be over- or underestimated. 

20 The absence of a clearly-defined common mode of action increases the level of 

21 uncertainty associated with the use of an RPF approach. It is not possible to determine whether 

22 cancer risks would be under- or overestimated by using a PAH RPF approach that assumes a 

23 common mode of action. The assumption of dose additivity inherent in the RPF approach may 

24 not be valid for a class of chemicals for which varying mechanisms of action occur to produce a 

25 common effect. A response addition methodology would be used to assess the combined risks 

26 from compounds with distinct mechanisms of action. For subgroups of PAHs with a common 

27 mechanism of action, an integrated RPF with a response addition approach may be applicable 

28 (U.S. EPA, 2000). 

29 The assumption of additivity cannot be confirmed or refuted based on evidence available 

30 in the peer-reviewed literature. The experimental data relating to dose additivity for PAH 

31 carcinogenicity are discussed in Section 2.7. Based on the available data, it appears that risks 

32 may be generally additive for complex mixtures, while binary mixtures can exhibit antagonism, 

33 synergism, or additivity. The level of confidence in the RPF approach would be increased if 

34 additivity could be demonstrated experimentally, even with simple mixtures. This remains a 

35 significant uncertainty in the proposed RPF approach. 

36 

37 8.6. UNCERTAINTY IN EXTRAPOLATING RPFs ACROSS EXPOSURE ROUTES 
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1 The peer consultation workshop (U.S. EPA, 2002) also identified uncertainty in 

2 extrapolation of RPFs across exposure routes. As with the 1993 Provisional Guidance, RPFs 

3 proposed in this analysis are also based on in vivo bioassay data collected using various routes of 

4 administration (e.g., dermal, intraperitoneal, subcutaneous, intramammillary, intramuscular, or 

5 intravenous injection, as well as lung implantation, tracheal implantation, and transplacental 

6 exposure after subcutaneous injection). The proposed RPF approach considers each bioassay 

7 type equivalent for the purpose of determining relative potency to benzo[a]pyrene. 

8 Table 8-3 compares the average RPFs (calculated from raw numbers and rounded to one 

9 significant digit) based on tumor bioassay data for each PAH across exposure routes. Dermal 

10 studies are shown collectively as well as separated by study type (complete or initiation). 

11 
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Table 8-3. Comparisons among average tumor bioassay RPF values by 

exposure route and target organ 

Dermal 

Dermal 

complete 

Dermal 

initiation 

Intraperitoneal 

Intraperitoneal, 

target organ = 

lung 

Intraperitoneal, 

target organ = 

liver 

Lung 

implantation

PAH N Average N Average N Average N Average N Average N Average N Average 

AA 1 0.5 1 0.5 – – – – – – – – 1 0.2 

AC – – – – – – – – – – – – – – 

BaA 1 0.02 – – 1 0.02 3 0.3a 1 0.08 2 0.4 – – 

BbcAC 

(1,12-MBA) 1 0.05 – – 1 0.05 – – – – – – – – 

BbF 3 0.3 1 0.2 2 0.4 3b1

c

 1 0.4 1 2 1 0.1 

BeAC 4 0.7 – – 4 0.7 – – – – – – – – 

BghiP – – – – – – – – – – – – 1 0.009 

BjAC – – – – – – 1 60d 1 60 – – – – 

BjF 3 0.06 – – 3 0.06 5b0.6a 2 0.5 1 0.7 1 0.03 

BkF 2 0.02 – – 2 0.02 – – – – – – 1 0.03 

BlAC 4 4 – – 4 4 – – – – – – – – 

CH 7 0.1 – – 7 0.1 3 0.1a 1 0.1 2 0.2 1 0.04 

CPcdP 7 0.2 3 0.3 4 0.1 1 1

d

 1 1 – – – – 

CPdefC 2 0.3 – – 2 0.3 – – – – – – – – 

DBacA – – – – – – – – – – – – – – 

DBaeF 2 0.9 1 1 1 0.7 – – – – – – – – 

DBaeP 2 0.4 1 0.3 1 0.4 – – – – – – – – 

DBahA 2 1 – – 2 1 1 10d 1 10 – – 1 2 

DBahP 1 0.9 – – 1 0.9 – – – – – – – – 

DBaiP 2 0.6 1 0.7 1 0.5 – – – – – – – – 

DBalP 2 30 – – 2 30 1 30d 1 30 – – – – 

FA – – – – – – 10 0.08a 8 0.05 2 0.2 – – 

IP – – – – – – – – – – – – 1 0.07 

N23eP 1 0.3 – – 1 0.3 – – – – – – – – 

PH – – – – – – – – – – – – – – 

Pyr – – – – – – – – – – – – – – 

aNewborn mouse model. 

bNumber of intraperitoneal RPFs includes those calculated for combined lung and liver incidence; these 

are not included in numbers of RPFs with lung or liver tumors. 

c

Includes both newborn mouse and adult A/J mouse models. 

dAdult A/J mouse model. 

1 

2 Likewise, intraperitoneal studies are shown grouped as well as separated by target organ 

3 (lung and liver). In general, the table shows that RPFs calculated from lung implantation and 

4 dermal studies are similar, while RPFs calculated from intraperitoneal studies tend to be higher 

5 for most compounds. However, intraperitoneal RPFs for chrysene (CH) and dibenz[a,l]pyrene 

6 (DBalP) are similar to dermal RPFs for these compounds. 
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1 One possible explanation for the higher intraperitoneal RPFs calculated from newborn 

2 mouse assays (footnoted “a” in the table) might be that the newborn mouse is more sensitive to 

3 the carcinogenic action of PAHs than an adolescent or adult mouse. Likewise, the adult 

4 A/J mouse is considered to be particularly sensitive to PAH lung tumorigenicity (Nesnow et al., 

5 1995), which may result in higher RPFs with this model (in Table 8-3, the intraperitoneal RPFs 

6 based on the A/J mouse model are footnoted “d”). There is little information to evaluate whether 

7 the newborn mouse is more or less sensitive than the adult A/J mouse model. Only one 

8 compound, benzo[b]fluoranthene (BbF), had RPFs calculated from both newborn mouse and 

9 adult A/J mouse models; the newborn mouse RPF was 2, while the A/J mouse RPF was 0.4. In 

10 summary, it is not clear whether the intraperitoneal RPFs are higher than dermal or lung 

11 implantation RPFs due to route-specific differences or animal model differences in susceptibility.

12 Cross-route extrapolation of relative potency estimates is a necessary, though uncertain, 

13 aspect of the RPF approach. It is difficult to determine which of the available study types (e.g., 

14 dermal, intraperitoneal, intratracheal) is most predictive of potential risks from oral and 

15 inhalation exposure in humans. In order to prioritize bioassays by exposure route, data are 

16 needed on relative potencies through environmentally relevant exposure routes (oral, inhalation, 

17 dermal) with relative potencies based on experimental exposure routes. 

18 The inhalation RPF scheme used by the California EPA (2004) employed a hierarchy of 

19 bioassay data based on exposure route (inhalation studies were preferred followed by 

20 intratracheal or intrapulmonary instillation, oral administration, skin-painting, and subcutaneous 

21 or intraperitoneal injection). Apart from the obvious preference for exposure routes that targeted 

22 the respiratory tract (inhalation, intratracheal, intrapulmonary), the basis for prioritizing the other 

23 exposure routes is not evident. Pufulete et al. (2004), who were also focused on PAHs as air 

24 contaminants, suggested that the clearance of PAHs after intratracheal instillation may be similar 

25 to clearance after inhalation exposure. The authors acknowledged that the high concentrations of 

26 PAHs used in intratracheal and intrapulmonary instillation studies may lead to major differences 

27 in pharmacokinetics, compared with inhalation exposure (Pufulete et al., 2004). Nevertheless, 

28 the authors suggested that intratracheal instillation of low doses of PAHs might be an appropriate 

29 surrogate exposure model for assessing relative potency of inhalation exposure. It is important 

30 to note that no intratracheal instillation studies were identified in the search for studies from 

31 which to calculate RPFs; thus, the information provided by Pufulete et al. (2004) is not directly 

32 useful for suggesting route-specific RPFs. Pufulete et al. (2004) did not provide any specific 

33 information on the relevance of intrapulmonary administration (a route used in several of the 

34 bioassays used to calculate RPFs) to inhalation exposure. 

35 As noted by U.S. EPA (2004), cross-route extrapolation would be contraindicated if there 

36 were convincing toxicokinetic evidence that absorption of PAHs does not occur by one or more 

37 exposure routes. However, available data on the absorption of PAHs indicates that, in general, 

38 PAHs are readily absorbed via ingestion, inhalation, and dermal exposure routes; however, the 
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1 rate of uptake varies with route and other factors (e.g., matrix, intake of fats and oils) (ATSDR, 

2 1995). Evidence for absorption of PAHs through these routes includes measurement of PAH3 DNA adducts at sites distal from the route of entry, measurement of urinary metabolites, and 

4 radiotracer studies in animals (ATSDR, 1995). U.S. EPA (2004) indicated that demonstration of 

5 any degree of uptake for each of the routes of interest is sufficient to allow the qualitative 

6 judgment to apply the route-to-route extrapolation; thus, cross-route extrapolation is supported 

7 by current data on the bioavailability of PAHs across several exposure routes. 

8 U.S. EPA (1994, 2004) also noted that point-of-entry toxicity may be considered contrary 

9 evidence for cross-route extrapolation. With respect to PAHs, available information on this issue 

10 is both limited and mixed. The one inhalation bioassay of benzo[a]pyrene, which suffered from 

11 several methodological limitations, identified the upper respiratory tract as the site of tumor 

12 formation, suggesting a point-of-entry effect (Thyssen et al., 1981). Dermal bioassays of 

13 benzo[a]pyrene have generally evaluated only skin tumors, precluding their use in determining 

14 whether distal tumors are induced. A number of early oral cancer bioassays of benzo[a]pyrene 

15 suggested that tumor formation was limited to point-of-entry sites (Rigdon and Neal, 1969, 1966; 

16 Neal and Rigdon, 1967). More recent oral carcinogenicity bioassays comparing MGP residue 

17 (Weyand et al., 1995) or coal tar preparations (Culp et al., 1998; Gaylor et al., 1998) with 

18 benzo[a]pyrene showed significant differences in target organ distribution of tumors between 

19 benzo[a]pyrene and complex mixtures of PAHs. Benzo[a]pyrene-induced tumors were observed 

20 primarily at the point of contact (i.e., the forestomach), while MGP residue and coal tar produced 

21 tumors in the lung, liver, forestomach, skin, and other organs. Other PAHs (e.g., 

22 benzo[c]fluorine) are proposed to be responsible for tumors at distal sites such as the lung 

23 (Koganti et al., 2000; Culp et al., 1998). However, a recent gavage study in rats (Kroese et al., 

24 2001) demonstrated that oral exposure to benzo[a]pyrene could induce tumors at distal sites, 

25 including the liver and auditory canal. Tissue-specific differences in metabolic activation and 

26 DNA binding of PAHs may contribute to the observed differences in target organ sensitivity 

27 (Weyand and Wu, 1995; Culp and Beland, 1994). 

28 In summary, available information provides some support for cross-route extrapolation. 

29 Absorption of PAHs across oral, inhalation, and dermal routes is evident and, while many of the 

30 cancer bioassays of benzo[a]pyrene suggested tumor formation limited to the point-of-entry, at 

31 least one recent study (Kroese et al., 2001) suggests that tumors may also be induced at distal 

32 sites. Furthermore, there is evidence that other PAHs (e.g., benzo[c]fluorene) may induce 

33 tumors at distal sites after oral exposure to coal tar preparations (Koganti et al., 2000; Culp et al., 

34 1998). However, cross-route extrapolation of RPFs is a significant source of uncertainty in this 

35 approach. 

36 Another approach to the issue of route-to-route extrapolation would be to prefer RPFs 

37 derived from particular target tissues deemed relevant to the exposure route of interest. For 

38 example, RPFs based on lung tumor data might be preferred for use in inhalation risk 
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1 assessment. To examine whether lung tumor RPFs were consistent across routes, RPFs 

2 calculated from lung tumor potency in intraperitoneal studies (both newborn mouse and adult 

3 A/J mouse models) were compared with RPFs from lung implantation studies in Table 8-3. 

4 RPFs for both intraperitoneal-lung and lung implantation studies were available for only four 

5 compounds (BbF, BjF, CH, and DBahA); for each of these, the intraperitoneal lung tumor RPF 

6 exceeded the lung implantation RPF. No information assessing the concordance between lung 

7 tumor potency after intraperitoneal administration and inhalation cancer potency was identified 

8 in the literature. The use of the final RPFs derived in this analysis across all routes of exposure is

9 recommended given the information outlined above and in the absence of data to indicate 

10 otherwise.
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APPENDIX A. SECONDARY SOURCES REVIEWED FOR IDENTIFICATION OF 

PRIMARY LITERATURE 

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benzo[j]fluoranthene, chrysene, dibenzo[a,h]pyrene, dibenzo[a,i]pyrene, and 5-methyl chrysene by the oral route. 

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A-2 DRAFT – DO NOT CITE OR QUOTE 

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assessment for exposure to PAH. In: Fifteenth international symposium on polycyclic aromatic compounds: 

Chemistry, biology and environmental impact. Belgirate, Italy, 19-22 September 1995. Ispra, Joint Research Centre 

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hydrocarbons (PAH). Part 1: hazard identification and dose-response assessment. Ontario Ministry of Environment 

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environmental health risk associated with exposure to airborne mixtures of polycyclic aromatic hydrocarbons 

(PAHs). Chemosphere 32(4):639–648. 

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exposure levels (AELs) in drinking water. Regul Toxicol Pharmacol 9(3):273–283. 

Schneider, K; Roller, M; Kalberlah, F; et al. (2002) Cancer risk assessment for oral exposure to PAH mixtures. J 

Appl Toxicol 22(1):73–83. 

Sjogren, M; Ehrenberg, L; Rannug, U. (1996) Relevance of different biological assays in assessing initiating and 

promoting properties of polycyclic aromatic hydrocarbons with respect to carcinogenic potency. Mutat Res 

358(1):97–112. 

Slooff, W; Janus, JA; Matthijsen, AJCM; et al. (1989) Integrated criteria document PAHs (PDF includes addendum 

by Montizaan). Bilthoven, The Netherlands, National Institute of Public Health and the Environment (RIVM). 

SRC (Syracuse Research Corporation). (1993) Estimating cancer risk from exposure to PAHs: a relative potency 

approach. SRC TR-93-045. Draft report prepared for U.S. EPA, Environmental Criteria and Assessment Office, 

Cincinnati, OH. 

U.S. EPA (Environmental Protection Agency). (1990) Drinking water criteria document for polycyclic aromatic 

hydrocarbons. Cincinnati, Ohio, Office of Health and Environmental Assessment, Environmental Criteria and 

Assessment Office. 

U.S. EPA. (1993) Provisional guidance for quantitative risk assessment of polycyclic aromatic hydrocarbons. 

Cincinnati, Ohio, Office of Health and Environmental Assessment, Environmental Criteria and Assessment Office. 
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1 APPENDIX B. BIBLIOGRAPHY OF STUDIES WITHOUT BENZO[A]PYRENE AS A 

2 REFERENCE COMPOUND 

3 

4 

5 

6 
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Table B-1. Bioassays with and without benzo[a]pyrene by PAH 

Bioassays with benzo[a]pyrene Bioassays without benzo[a]pyrene 

Dermal Dermal 

PAH CASRN Initiation Complete 

Intraperitoneal Sub- cutaneous Oral Other Initiation Complete Intra- peritoneal Sub- cutaneous Oral Oth

Aceanthrylene 202-03-09 Acenaphthene 83-32-9 Acenaphthylene 208-96-8 Acephenanthrylene 201-06-9 Acepyrene, 2,3- 25732-74-5 x x Anthanthrene 191-26-4 x x x x x Anthracene 120-12-7 x x x x x x x x Benz[a]anthracene 56-55-3 x x x x x x x x x x x Benz[b]anthracene 92-24-9 Benz[b,c]aceanthrylene, 11H- 202-94-8 x Benz[e]aceanthrylene 199-54-2 Benz[j]aceanthrylene 202-33-5 x x Benz[l]aceanthrylene 211-91-6 x Benzacenaphthylene 76774-50-0 Benzo[a]fluoranthene 203-33-8 x Benzo[a]fluorene 

238-84-6 or 

30777-18-5 x 

Benzo[a]perylene 191-85-5 Benzo[b]chrysene 214-17-5 x Benzo[b]fluoranthene 205-99-2 x x x x x x 11H-Benzo[b]fluorene 

243-17-4 or 

30777-19-6 x 

Benzo[b]perylene 197-70-6 Benzo[c]chrysene 194-69-4 Benzo[c]fluorene 

205-12-9 or 

30777-20-9 x 

Benzo[c]phenanthrene 195-19-7 x x x x Benzo[e]pyrene 192-97-2 x x x x x Benzo[g]chrysene 196-78-1 Benzo[g,h,i]fluoranthene 203-12-3 x x Benzo[g,h,i]perylene 191-24-2 x x x x Benzo[j]fluoranthene 205-82-3 x x x x x x Benzo[k]fluoranthene 207-08-9 x x x x x Benzophenanthrene 65777-08-4 Chrysene 218-01-9 x x x x x x x x x Coronene 191-07-1 x x Cyclopenta[c,d]pyrene 27208-37-3 x x x x Cyclopenta[d,e,f]chrysene, 4H- 202-98-2 x x Cyclopenta[d,e,f]phenanthrene, 4H- 203-64-5 Cyclopenta[h,i]acephenanthrylene 114959-37-4 
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Table B-1. Bioassays with and without benzo[a]pyrene by PAH 

Bioassays with benzo[a]pyrene Bioassays without benzo[a]pyrene 

Dermal Dermal 

PAH CASRN Initiation Complete 

Intraperitoneal Sub- cutaneous Oral Other Initiation Complete Intra- peritoneal Sub- cutaneous Oral Oth

Cyclopenta[h,i]aceanthrylene 131581-33-4 Cyclopentaphenanthrene 219-08-9 Cyclopenteno-1,2-benzanthracene, 5,6- 7099-43-6 x Dibenz[a,c]anthracene 215-58-7 x x x x x x Dibenzo[a,e]fluoranthene 5385-75-1 x x x Dibenz[a,j]anthracene 224-41-9 x Dibenzo[b,e]fluoranthene 2997-45-7 Dibenzo[a,c]fluorene, 13H- 201-65-0 Dibenzo[a,e]pyrene 192-65-4 x x x Dibenzo[a,f]fluoranthene 203-11-2 x x x x Dibenzo[a,g]fluorene, 13H- 207-83-0 x Dibenz[a,h]anthracene 53-70-3 x x x x x x x x x x x Dibenzo[a,h]pyrene 189-64-0 x x x x Dibenzo[a,i]pyrene 189-55-9 x x x x x x Dibenzo[a,l]pyrene 191-30-0 x x x x x x x xDibenzo[e,l]pyrene 192-51-8 x x Dibenzo[h,rst]pentaphene 192-47-2 Dibenz[k,mno]acephenanthrylene 153043-81-3 Dibenzo[j,mno]acephenanthrylene 153043-82-4 Dihydroaceanthrylene, 1,2- 641-48-5 x Fluoranthene 206-44-0 x x x x Fluorene 86-73-7 x x Indeno[1,2,3-c,d]fluoranthene 193-43-1 Indeno[1,2,3-c,d]pyrene 193-39-5 x x x x x Naphtho[1,2-b]fluoranthene 111189-32-3 x 

Naphtho[1,2,3,-mno]acephenanthrylene 113779-16-1 

Naphtho[2,1-a]fluoranthene 203-20-3 x 

Naphtho[2,3-a]pyrene 196-42-9 Naphtho[2,3-e]pyrene 193-09-9 x x 

Pentacene 135-48-8 Pentaphene 222-93-5 Perylene 198-55-0 x x x Phenanthrene 85-01-8 x x x x x x x x x x Picene 213-46-7 x x x x Pyrene 129-00-0 x x x x x Tribenzofluoranthene 3,4-10,11-12,13- 13579-05-0 Triphenylene 217-59-4 x PAHs in bold have at least one bioassay without BaP and no bioassays with BaP. 

1 
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B.1. BIBLIOGRAPHY OF BIOASSAYS WITHOUT BENZO[A]PYRENE 

Amin, S; Huie, K; Hecht, SS; (1985) Mutagenicity and tumor-initiating activity of methylated 

benzo[b]fluoranthenes. Carcinogenesis 6:1023–1025.

Amin, S; Hussain, N; Balanikas, G; et al. (1985) Mutagenicity and tumor initiating activity of methylated benzo[k] 

fluoranthenes. Cancer Lett 26:343–347. 

Amin, S; Misra, B; Braley, J; et al. (1991) Comparative tumorigenicity in newborn mice of chrysene and

5-alkylchrysene-1,2-diol-3,4-epoxides. Cancer Lett 58:115–118. 

Amin, S; Weyand, EH; Huie, K; et al. (1991) Effects of fluorine substitution on benzo[b]fluoranthene tumorgenicity 

and DNA adduct formation in mouse skin. In: Cooke, M; Loening, K; Merritt, J, eds. Polynuclear aromatic 

hydrocarbons: Measurements, means and metabolism. Columbus, OH, Battelle Press pp. 25–35. 

Amin, S; Desai, D; Dai, W; et al. (1995) Tumorigenicity in newborn mice of fjord region and other sterically 

hindered diol epoxides of benzo[g]chrysene, dibenzo[a,l]pyrene (dibenzo[def,p]chrysene), 

4H-cyclopenta[def]chrysene and fluoranthene. Carcinogenesis 16:2813–2817. 

Barry, G; Cook, CW; Amin, S; et al. (1934) A comparison of the action of some polycyclic aromatic hydrocarbons 

in producing tumours of connective tissue. Am J Cancer 20:58–69. 

Bhatt, TS; Coombs, MM. (1990) The carcinogenicity of cyclopenta[a]phenanthrene and chrysene derivatives in the 

Sencar mouse. Polycyclic Aromat Compd 1:51–58. 

Bock, FG; King, DW. (1959) A study of the sensitivity of the mouse forestomach toward certain polycyclic 

hydrocarbons. J Natl Cancer Inst 23:833–838. 

Bottomley, AC; Twort, CC. (1934) The carcinogenicity of chrysene and oleic acid. Am J Cancer 21:781–786. 

Boyland, E; Burrows, H. (1935) The experimental production of sarcoma in rats and mice by a colloidal aqueous 

solution of 1:2:5:6-dibenzanthracene. J Pathol Bacteriol 41:231–238. 

Boyland, E; Sims, P. (1967) The carcinogenic activities in mice of compounds related to benz[a]anthracene. Int J 

Cancer 2:500–504. 

Buening, MK; Levin, W; Karle, JM; et al. (1979) Tumorigenicity of bay-region epoxides and other derivatives of 

chrysene and phenanthrene in newborn mice. Cancer Res 39:5063–5068. 

Buening, MK; Levin, W; Wood, A; et al. (1979) Tumorigenicity of the dihydrodiols of dibenz[a,h]anthracene on 

mouse skin and in newborn mice. Cancer Res 39:1310–1314. 

Buters, JT; Mahadevan, B; Quintanilla-Martinez, L; et al. (2002) Cytochrome P450 1B1 determines susceptibility to 

dibenzo[a,l]pyrene-induced tumor formation. Chem Res Toxicol 15:1127–1135. 

Cavalieri, EL; Rogan, EG; Higginbotham, S; et al. (1989) Tumor-initiating activity in mouse skin and 

carcinogenicity in rat mammary gland of dibenzo[a]pyrenes: the very potent environmental carcinogen dibenzo[a, 

l]pyrene. J Cancer Res Clin Oncol 115:67–72. 

Chang, RL; Levin, W; Wood, AW; et al. (1981) Tumorigenicity of the diastereomeric bay-region benzo[e]pyrene 

9,10-diol-11,12-epoxides in newborn mice. Cancer Res 41:915–918. 

Chang, RL; Levin, W; Wood, AW; et al. (1982) Tumorigenicity of bay-region diol-epoxides and other benzo-ring 

derivatives of dibenzo[a,h]pyrene and dibenzo[a,i]pyrene on mouse skin and in newborn mice. Cancer Res 42:25–

29. 
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Chang, RL; Levin, W; Wood, AW; et al. (1983) Tumorigenicity of enantiomers of chrysene 1,2-dihydrodiol and of 

the diastereomeric bay-region chrysene 1,2-diol-3,4-epoxides on mouse skin and in newborn mice. Cancer Res 

43:192–196. 

Chouroulinkov, I; Coulomb, H; MacNicoll, AD; et al. (1983) Tumour-initiating activities of dihydrodiols of 

dibenz[a,c]anthracene. Cancer Lett 19:21–26. 

Danz, M; Hartmann, A; Otto, M; et al. (1991) Hitherto unknown additive growth effects of fluorene and 

2-acetylaminofluorene on bile duct epithelium and hepatocytes in rats. Arch Toxicol Suppl 14:71–74. 

Flesher, JW; Horn, J; Lehner, AF. (2002) Comparative carcinogenicity of picene and dibenz[a,h]anthracene in the 

rat. Biochem Biophys Res Commun 290:275–279. 

Forbes, PD; Davies, RE; Urbach, F. (1976) Phototoxicity and photocarcinogenesis: Comparative effects of 

anthracene and 8-methoxypsoralen in the skin of mice. Food Cosmet Toxicol 14:303–306. 

Geddie, JE; Amin, S; Huie, K; et al. (1987) Formation and tumorigenicity of benzo[b]fluoranthene metabolites in 

mouse epidermis. Carcinogenesis 8:1579–1584. 

Gill, HS; Kole, PL; Wiley, JC; et al. (1994) Synthesis and tumor-initiating activity in mouse skin of 

dibenzo[a,l]pyrene syn- and anti-fjord-region diolepoxides. Carcinogenesis 15:2455–2460. 

Hecht, SS; LaVoie, E; Amin, S; et al. (1980) On the metabolic activation of the benzofluoranthenes. Polynucl Arom 

Hydrocarb 417–433. 

Hecht, SS; LaVoie, EJ; Bedenko, V; et al. (1981a) On the metabolic activation of dibenzo[a,i]pyrene and dibenzo 

[a,h]pyrene. Chem Analy & Biol Fate: Polynucl Arom Hydrocarb 43–45. 

Hecht, SS; LaVoie, EJ; Bedenko, V; et al. (1981b) Reduction of tumorigenicity and of dihydrodiol formation by 

fluorine substitution in the angular rings of dibenzo[a,i]pyrene. Cancer Res 41:4341–4345. 

Hecht, SS; Amin, S; Lin, JM; et al. (1995) Mammary carcinogenicity in female CD rats of a diol epoxide metabolite 

of fluoranthene, a commonly occurring environmental pollutant. Carcinogenesis 16:1433–1435. 

Hecht, SS; Rivenson, A; Amin, S; et al. (1996) Mammary carcinogenicity of diol epoxide metabolites of 

benzo[j]fluoranthene in female CD rats. Cancer Lett 106:251–255. 

Hermann, M. (1981) Synergistic effects of individual polycyclic aromatic hydrocarbons on the mutagenicity of their 

mixtures. Mutat Res 90:399–409. 

Hill, WT; Stanger, DW; Pizzo, A; et al. (1951) Inhibition of 9,10-dimethyl-1,2-benzanthracene skin carcinogenesis 

in mice by polycyclic hydrocarbons. Cancer Res 11:892–897. 

Homburger, F; Treger, A. (1970) Transplantation technique for acceleration of carcinogenesis by benz[a]anthracene 

or 3,4,9,10-dibenzpyrene. J Natl Cancer Inst 44:357–360. 

Homburger, F; Treger, A; Boger, E. (1971) Inhibition of murine subcutaneous and intravenous 

benzo[rst]pentaphene. Carcinogenesis by sweet orange oils and d-limonene. Oncology 25:1–10. 

Jerina, DM; Sayer, JM; Yagi, H; et al. (1981) Highly tumorigenic bay-region diol epoxides from the weak 

carcinogen benzo[c]phenanthrene. Adv Exp Med Biol 136 Pt A:501–523. 

Johnson, S. (1968) Effect of thymectomy on the induction of skin tumours by dibenzanthracene, and of breast 

tumours by dimethylbenzanthracene in mice of the IF strain. Br J Cancer 22:755–761. 

Klein, M. (1952) Effect of croton oil on induction of tumors by 1,2-benzanthracene, deoxychloric or low doses of 

20-methylcholanthrene in mice. J Nat1 Cancer Inst 13:333–341. 
B-6 DRAFT – DO NOT CITE OR QUOTE 

Klein, M. (1960) A comparison of the initiating and promoting actions of 9,10-dimethyl-1,2-benzanthracene and 

1,2,5,6-dibenzanthracene in skin tumorigenesis. Cancer Res 20:1179–1183. 

Klein, M. (1963) Susceptibility of strain B6AF1/J hybrid infant mice to tumorigenesis with 1,2-benzanthracene, 

deoxycholic acid, and 3-methylcholanthrene. II. Tumours called forth by painting the skin with dibenzpyrene. 

Cancer Res 23:1701–1707. 

Kouri, RE; Connolly, GM; Nebert, DW; et al. (1983) Association between susceptibility to dibenzanthracene 

induced fibrosarcoma formation and the Ah locus. Int J Cancer 32:765–768. 

Lacassagne, A; Buu-Hoi, NP; Zajdela, F; et al. (1968) The true dibenzo[a,l]pyrene, a new, potent carcinogen. 

Naturwissenschaften 55:43. 

LaVoie, EJ; Tulley L; Bedenko, V; et al. (1980) Mutagenicity, tumor initiating activity, and metabolism of tricyclic 

polynuclear aromatic hydrocarbons. In: Bjorseth, A; Dennis, AJ, eds. Polynuclear aromatic hydrocarbons: 

Chemistry and biological effects. Columbus, OH: Battelle Press; pp. 1041–1057. 

LaVoie, EJ; Tulley-Freiler, L; Bedenko, V; et al. (1981) Comparative studies on the tumor initiating activity and 

metabolism of methylfluorenes and methylbenzofluorenes. In: Cooke, M; Dennis, AJ, eds. Chemical analysis and 

biological fate: Polynuclear hydrocarbons. Columbus, OH: Battelle Press; pp. 417–427. 

LaVoie, EJ; Coleman, DT; Tonne, RL; et al. (1983) Mutagenicity, tumor initiating activity and metabolism of 

methylated anthracenes. In: Cooke, M; Dennis, AJ, eds. Proceedings of the Seventh International Symposium. 

Columbus, OH: Battelle Press pp. 785–798. 

LaVoie, EJ; Cai, ZW; Meegalla, RL; et al. (1993a) Evaluation of the tumor-initiating activity of 4-, 5-, 6-, and 

7-fluorobenzo[b]fluoranthene in mouse skin. Chem Bio Interact 89:129–139. 

LaVoie, EJ; He, ZM; Meegalla, RL; et al. (1993b) Exceptional tumor-initiating activity of 4-fluorobenzo[j]-

fluoranthene on mouse skin: comparison with benzo[j]-fluoranthene, 10-fluoro-benzo[j]fluoranthene, 

benzo[a]pyrene, dibenzo[a,l]pyrene and 7,12-dimethylbenz[a]anthracene. Cancer Lett 70:7–14. 

LaVoie, EJ; He, ZM; Wu, Y; et al. (1994) Tumorigenic activity of the 4,5- and 9,10-dihydrodiols of 

benzo[j]fluoranthene and their syn- and anti-diol epoxides in newborn mice. Cancer Res 54:962–968. 

Levin, W; Wood, AW; Chang, RL; et al. (1978) Evidence for bay region activation of chrysene 1,2-dihydrodiol to 

an ultimate carcinogen. Cancer Res 38:1831–1834. 

Levin, W; Wood, AW; Chang, RL; et al. (1980) Exceptionally high tumor-initiating activity of 

benzo[c]phenanthrene bay-region diol-epoxides on mouse skin. Cancer Res 40:3910–3914. 

Levin, W; Chang, RL; Wood, AW; et al. (1984) High stereoselectivity among the optical isomers of the 

diastereomeric bay-region diolepoxides of benz[a]anthracene in the expression of tumorigenic activity in murine 

tumor models. Cancer Res 44:929–933. 

Levin, W; Chang, RL; Wood, AW; et al. (1986) Tumorigenicity of optical isomers of the diastereomeric bay-region 

3,4-diol-1,2-epoxides of benzo[c]phenanthrene in murine tumor models. Cancer Res 46:2257–2261. 

Lijinsky, W; Garcia, H. (1972) Skin carcinogenesis tests of hydrogenated derivatives of anthanthrene and other 

polynuclear hydrocarbons. Z Krebsforsch 77:226–230. 

Lijinsky, WH; Garcia, B; Terrracini, B. (1965) Tumorigenic activity of hydrogenated derivatives of 

dibenz[a,h]anthracene. J Natl Cancer Inst 34:1–6. 

Lijinsky, W; Garcia, H; Saffiotti, U. (1970) Structure-activity relationships among some polynuclear hydrocarbons 

and their hydrogenated derivatives. J Natl Cancer Inst 44:641–649. 

Lorenz, E; Stewart, HL. (1947) Tumors of the alimentary tract induced in mice by feeding olive oil emulsions 

containing carcinogenic hydrocarbons. J Natl Cancer Inst 7:227–238. 
B-7 DRAFT – DO NOT CITE OR QUOTE 

Lorenz, E; Stewart, HL. (1948) Tumors of alimentary tract in mice fed carcinogenic hydrocarbons in mineral-oil 

emulsions. J Natl Cancer Inst 9:173–180. 

Lubet, RA; Connolly, GM; Nebert, DW; et al. (1983) Dibenz[a,h]anthracene-induced subcutaneous tumors in mice. 

Strain sensitivity and the role of carcinogen metabolism. Carcinogenesis 4:513–517. 

Malament, DS; Shklar, G. (1981) Inhibition of DMBA carcinogenesis of hamster buccal pouch by phenanthrene and 

dimethylnaphthalene. Carcinogenesis 2:723–729. 

Mass, MJ; Abu-Shakra, A; Roop, BC; et al. (1996) Benzo[b]fluoranthene: tumorigenicity in strain A/J mouse lungs, 

DNA adducts and mutations in the Ki-ras oncogene. Carcinogenesis 17:1701–1704. 

Nakatsuru, Y; Wakabayashi, K; Fujii-Kuriyama, Y; et al. (2004) Dibenzo[a,l]pyrene-induced genotoxic and 

carcinogenic responses are dramatically suppressed in aryl hydrocarbon receptor-deficient mice. Int J Cancer 

112:179–183. 

Nesnow, S; Gold, A; Sangaiah, R; et al. (1993) Mouse skin tumor-initiating activity of benz[j]aceanthrylene in 

SENCAR mice. Cancer Lett 73:73–76. 

Nesnow, S; Ross, JA; Nelson, G; et al. (1994) Cyclopenta[cd]pyrene-induced tumorigenicity, Ki-ras codon 12 

mutations and DNA adducts in strain A/J mouse lung. Carcinogenesis 15:601–606. 

O'Gara, RW; Kelly, MG; Brown, J; et al. (1965) Induction of tumors in mice given a minute single dose of 

dibenz[a,h]anthracene or 3-methylcholanthrene as newborns: A dose-response study. J Natl Cancer Inst 

35(6):1027–1042. 

Platt, KL; Pfeiffer, EH; Glatt, HR; et al. (1983) Bacterial mutagenicity and carcinogenicity of potential metabolites 

of dibenz[a,h]anthracene. J Cancer Res Clin Oncol 105:A23. 

Platt, KL; Pfeiffer, E; Petrovic, P; et al. (1990) Comparative tumorigenicity of picene and dibenz[a,h]anthracene in 

the mouse. Carcinogenesis 11:1721–1726. 

Platt, KL; Dienes, HP; Tommasone, M; et al. (2004) Tumor formation in the neonatal mouse bioassay indicates that 

the potent carcinogen dibenzo[def,p]chrysene (dibenzo[a,l]pyrene) is activated in vivo via its 

trans-11,12-dihydrodiol. Chem Biol Interact 148:27–36. 

Pollia, JA. (1939) Investigations on the possible carcinogenic effect of anthracene and chrysene and some of their 

compounds. I. The effect of skin painting on the skin of mice. J Ind Hyg Toxicol 23:449–451. 

Pollia, JA. (1941) Investigation on the possible carcinogenic effect of anthracene and chrysene and some of their 

compounds. II. The effect of subcutaneous injection in rats. J Ind Hyg Toxicol 223:449–451. 

Prahalad, AK; Ross, JA; Nelson, GB; et al. (1997) Dibenzo[a,l]pyrene-induced DNA adduction, tumorigenicity, and 

Ki-ras oncogene mutations in strain A/J mouse lung. Carcinogenesis 18:1955–1963. 

Ranadive, KJ; Karande, KA. (1963) Studies on 1,2,5,6-dibenzanthracene-induced mammary carcinogenesis in mice. 

Br J Cancer 17:272–280. 

Rice, JE; Coleman, DT; Hosted, TJJ; et al. (1985) On the metabolism, mutagenicity, and tumor-initiating activity of 

indeno[1,2,3-cd]pyrene. pp. 1097–1109. 

Rice, JE; Hosted, TJ Jr; DeFloria, MC; et al. (1986) Tumor-initiating activity of major in vivo metabolites of 

indeno[1,2,3-cd]pyrene on mouse skin. Carcinogenesis 7:1761–1764. 

Rice, JE; Weyand, EH; Geddie, NG; et al. (1987) Identification of tumorigenic metabolites of benzo[j]fluoranthene 

formed in vivo in mouse skin. Cancer Res 47:6166–6170. 
B-8 DRAFT – DO NOT CITE OR QUOTE 

Rice, JE; Weyand, EH; Burrill, C; et al. (1990) Fluorine probes for investigating the mechanism of activation of 

indeno[1,2,3-cd]pyrene to a tumorigenic agent. Carcinogenesis 11:1971–1974. 

Riegel, B; Watman, WB; Hill, WT. (1951) Delay of methylcholanthrene skin carcinogenesis in mice by 

1,2,5,6-dibenzofluorene. Cancer Res 11:301–306. 

Salaman, MH; Roe, FJC. (1956) Further tests for tumour-initiating activity: N,N-di(2-chloroethyl)- 

paminophenylbutic acid (CB1348) as an initiator of skin tumour formation in the mouse. Br J Cancer 10:363–378. 

Sardella, DJ; Boger, E; Ghoshal, PK. (1981) Active sites in hexacyclic carcinogens probed by the fluorine 

substitution methodology. Columbus, OH: Battelle Press; pp. 529–538. 

Schmähl, D. (1955) [Testing of naphthalene and anthracene for carcinogenic effects in rats.] (in German). Z 

Krebsforsch 60:697–710. 

Schoental, R. (1959) Carcinogenic activity of 3:4:9:10-dibenzopyrene. Acta Unio Int Contra Cancrum 15:216–219. 

Schoket, B; Hewer, A; Grover, PL; et al. (1988) Covalent binding of components of coal-tar, creosote and bitumen 

to the DNA of the skin and lungs of mice following topical application. Carcinogenesis 9:1253–1258. 

Scribner, JD. (1973) Brief communication: Tumor initiation by apparently noncarcinogenic polycyclic aromatic 

hydrocarbons. J Natl Cancer Inst 50:1717–1719. 

Sellakumar, A; Shubik, P. (1974) Carcinogenicity of different polycyclic hydrocarbons in the respiratory tract of 

hamsters. J Natl Cancer Inst 53:1713–1719. 

Shear, MJ. (1938) Studies in carcinogenesis. V. Methyl derivatives of 1,2-benzanthracene. Am J Cancer 33:499–

537. 

Shear, MJ; Leiter, J. (1941) Studies in carcinogenesis. XVI. Production of subcutaneous tumors in mice by 

miscellaneous polycyclic compounds. J Natl Cancer Inst 2:241–259. 

Siebert, D; Marquardt, H; Friesel, H; et al. (1981) Polycyclic aromatic hydrocarbons and possible metabolites: 

Convertogenic activity in yeast and tumor initiating activity in mouse skin. J Cancer Res Clin Oncol 102:127–139. 

Slaga, TJ; Gleason, GL; Mills, C; et al. (1980) Comparison of the tumour-initiating activities of dihydrodiols and 

diol-epoxides of various polycyclic aromatic hydrocarbons. Cancer Res 40:1981–1984. 

Snell, KC; Stewart, HL. (1962) Pulmonary adenomatosis induced in DBA/2 mice by oral administration of 

dibenz(a,h]anthracene. J Natl Cancer Inst 28:1043–1049. 

Snell, KC; Stewart, HL. (1963) Induction of pulmonary adenomatoses in DBA/2 mice by the oral administration of 

dibenz[a,h]anthracene. Acta Un Int Cancer 19:692–694. 

Stanton, MF; Miller, E; Wrench, C; et al. (1972) Experimental induction of epidermoid carcinoma in the lungs of 

rats by cigarette smoke condensate. J Nat1 Cancer Inst 49:867–877. 

Steiner, PE; Edgcomb, JH. (1952) Carcinogenicity of 1,2-benzanthracene. Cancer Res 12:657–659. 

Steiner, PF; Falk, HL. (1951) Summation and inhibition effects of weak and strong carcinogenic hydrocarbons: 

1:2-Benzanthracene, chrysene, 1:2:5:6-dibenzanthracene, and 20-methylcholanthrene. Cancer Res 11:56–63. 

Stenbk, F; Sellakumar, A. (1974) Lung tumor induction by dibenz(a,i]pyrene in the Syrian golden hamster. Z 

Krebsforsch 82:175–182. 

Stevenson, JL; Von Haam, E. (1965) Carcinogenicity of benz[a]anthracene and benzo[c]phenanthrene derivatives. 

Am Ind Hyg Assoc J 26:475–478. 
B-9 DRAFT – DO NOT CITE OR QUOTE 

Tawfic, HN. (1965) Studies on ear duct tumors in rats. Part II: Inhibitory effect of methylcholanthrene and 

1,2-benzanthracene on tumor formation by 4-dimethylamino-stilbene. Acta Pathol Jpn 15:255–260. 

Van Duuren, BL; Langseth, L; Goldschmidt, BM. (1967) Carcinogenicity of epoxides, lactones and peroxy 

compounds: VI. Structure and carcinogenic activity. J Nat1 Cancer Inst 39:1217–1227. 

Van Duuren, BL; Sivak, A; Langseth, L; et al. (1968) Initiators and promoters in tobacco carcinogenesis. Natl 

Cancer Inst Monogr 28:173–180. 

Van Duuren, BL; Sivak, A; Goldschmidt, BM; et al. (1970) Initiating activity of aromatic hydrocarbons in two-stage 

carcinogenesis. J Natl Cancer Inst 44:1167–1173. 

Vulimiri, SV; Baer-Dubowska, W; Harvey, RG; et al. (1999) Analysis of highly polar DNA adducts formed in 

SENCAR mouse epidermis following topical application of dibenz[a,j]anthracene. Chem Res Toxicol 12:60–67. 

Wang, JS; Busby, WF. Jr. (1993) Induction of lung and liver tumors by fluoranthene in a preweanling CD-1 mouse 

bioassay. Carcinogenesis 14:1871–1874. 

Waravdekar, SS; Ranadive, KJ. (1958) Biologic testing of 3,4,9,10-dibenzpyrene. J Natl Cancer Inst 21:1151–1159. 

Weyand, EH; Amin, S; Huie, K; et al. (1989) Effects of fluorine substitution on the DNA binding and 

tumorigenicity of benzo[b]fluoranthene in mouse epidermis. Chem Biol Interact 71:279–290. 

Weyand, EH; Patel, S; LaVoie, EJ; et al. (1990) Relative tumor initiating activity of benzo[a]fluoranthene, 

benzo[b]fluoranthene, naphtho[1,2-b]fluoranthene and naphtho[2,1-a]fluoranthene on mouse skin. Cancer Lett 

52:229–233. 

Weyand, EH; Cai, ZW; Wu, Y; et al. (1993) Detection of the major DNA adducts of benzo[b]fluoranthene in mouse 

skin: role of phenolic dihydrodiols. Chem Res Toxicol 6:568–577. 

White, FR; Eschenbrenner, AB. (1945) Note on the occurrence of hepatomas in rats following the ingestion of 

1,2-benzoanthracene. J Natl Cancer Inst 6:19–21. 

Wislocki, PG; Buening, MK; Levin, W; et al. (1979) Tumorigenicity of the diastereomeric benz[a]anthracene 

3,4-diol-1,2-epoxides and the (+)-and (-)-enantiomers of benz [a]anthracene 3,4-dihydrodiol in newborn mice. J 

Natl Cancer Inst 63:201–204. 

Wodinsky, I; Helinski, A; Kensler, CJ. (1964) Susceptibility of Syrian hamsters to induction of fibrosarcomas with a 

single injection of 3,4,9,10-dibenzpyrene. Nature 203:308–309. 

Wood, AW; Chang, RL; Levin, W; et al. (1979) Mutagenicity and tumorigenicity of phenanthrene and chrysene 

epoxides and diol epoxides. Cancer Res 39:4069–4077. 

Wynder, EL.; Hoffmann, D. (1961) Carcinogenicity of dibenzo (a,1]pyrene. Nature 192:1092–1093. 

Zajdela, F; Perin-Roussel, O; Saguem, S. (1987) Marked differences between mutagenicity in Salmonella and 

tumour-initiating activities of dibenzo[a,e]fluoranthene proximate metabolites; initiation inhibiting activity of 

norharman. Carcinogenesis 8:461–464. 
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B.2. BIBLIOGRAPHY OF STUDIES ON CANCER-RELATED ENDPOINTS 

WITHOUT BENZO[A]PYRENE 

Abe, S; Sasaki, M. (1977) Studies on chromosomal aberrations and sister chromatid exchanges induced by

chemicals. Proc Jpn Acad 53:46–49. 

Agarwal, SK; Sayer, JM; Yeh, HJC; et al. (1987) Chemical characterization of DNA adducts derived from the 

configurationally isomeric benzo[c]phenanthrene-3,4-diol 1,2-epoxides. J Am Chem Soc 109:2497–2504. 

Agarwal, R; Canella, KA; Yagi, H; et al. (1996) Benzo[c]phenanthrene-DNA adducts in mouse epidermis in relation 

to the tumorigenicities of four configurationally isomeric 3,4-dihydrodiol 1,2-epoxides. Chem Res Toxicol 9:586–

592. 

Agarwal, R; Coffing, SL; Baird, WM; et al. (1997) Metabolic activation of benzo[g]chrysene in the human 

mammary carcinoma cell line MCF-7. Cancer Res 57:415–419. 

Amin, S; Desai, D; Hecht, SS. (1993) Tumor-initiating activity on mouse skin of bay region diol-epoxides of 

5,6-dimethylchrysene and benzo[c]phenanthrene. Carcinogenesis 14:2033–2037. 

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B-18 DRAFT – DO NOT CITE OR QUOTE 

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B-19 DRAFT – DO NOT CITE OR QUOTE 

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B-20 DRAFT – DO NOT CITE OR QUOTE 

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153. 
C-1 DRAFT – DO NOT CITE OR QUOTE 

1 APPENDIX C. DOSE-RESPONSE DATA FOR POTENCY CALCULATIONS 

2 

3 
C-2 DRAFT – DO NOT CITE OR QUO

Table C-1. Dermal bioassays: dose response information for incidence data 

Record 

number Reference 

Study 

type Species 

Tumor 

type PAH Sex

Dose 

of 

PAH

Dose 

units 

Number of 

animals 

with tumors

Number 

of 

animals 

in group

% tumorbearing 

animals 

Results of 

authors' 

statistical 

analysis 

(p-value)

Fisher's 

exact 

p-value 

CochranArmitage 

trend test 

p-value Com

Complete carcinogenicity studies 

600 Habs et al., 

1980 

Complete Mice Sum of 

Papilloma, carcinoma, sarcoma 

Acetone F 0 pg/animal 0 35 0% 

 DMSO F 0 pg/animal 0 36 0%  BaP F 1.7 pg/animal 8 34 24% 1.92 × 10-3 BaP F 2.8 pg/animal 24 35 69% 1.67 × 10-11 BaP F 4.6 pg/animal 22 36 61% 2.1 × 10-9 2.15 × 10-9 BbF F 3.4 pg/animal 2 38 5% 2.6 × 10-1 BbF F 5.6 pg/animal 5 34 15% 2.3 × 10-2 BbF F 9.2 pg/animal 20 37 54% 3.7 × 10-8 1.33 × 10-9 BjF F 3.4 pg/animal 1 38 3% 5.1 × 10-1 BjF F 5.6 pg/animal 1 35 3% 4.9 × 10-1 BjF F 9.2 pg/animal 2 38 5% 2.6 × 10-1 1.77 × 10-1 BkF F 3.4 pg/animal 1 39 3% 5.2 × 10-1 BkF F 5.6 pg/animal 0 38 0%  BkF F 9.2 pg/animal 0 38 0%  CPcdP F 1.7 pg/animal 0 34 0%  CPcdP F 6.5 pg/animal 0 35 0%  CPcdP F 27.2 pg/animal 3 38 8% 1.3 × 10-1 6.36 × 10-2 IP F 3.4 pg/animal 1 36 3% 5 × 10-1 IP F 5.6 pg/animal 0 37 0%  IP F 9.2 pg/animal 0 37 0%  CO F 5.6 pg/animal 1 39 3% 0.52  CO F 15 pg/animal 2 40 5% 0.27 1.83 × 10-113640 Cavalieri et 

al., 1983 

Complete Mice Papilloma, 

adenoma, carcinoma 

Acetone F 0 nmol 0 29 0% 

 BaP F 2.2 nmol 2 30 7% 0.25  BaP F 6.6 nmol 2 28 7% 0.24 
C-3 DRAFT – DO NOT CITE OR QUO

Table C-1. Dermal bioassays: dose response information for incidence data 

Record 

number Reference 

Study 

type Species 

Tumor 

type PAH Sex

Dose 

of 

PAH

Dose 

units 

Number of 

animals 

with tumors

Number 

of 

animals 

in group

% tumorbearing 

animals 

Results of 

authors' 

statistical 

analysis 

(p-value)

Fisher's 

exact 

p-value 

CochranArmitage 

trend test 

p-value Com

 BaP F 20 nmol 17 30 57% 4.32 × 10-7 2.96 × 10-1 CPcdP F 22.2 nmol 2 29 7% 0.25  CPcdP F 66.6 nmol 2 29 7% 0.25  CPcdP F 200 nmol 24 29 83% 9.25 × 10-12 1.39 × 10-16620 Hoffmann 

and Wynder 1966 

Complete Mice Papillomas Dioxane F 0 % 0 20 0% 

 BaP F 0.05 % 17 20 85% 1.28 × 10-8 BaP F 0.1 % 19 20 95% 1.5 × 10-10 8.7 × 10-10 DBaeP F 0.05 % 16 30 53% 3.31 × 10-5 DBaeP F 0.1 % 9 17 53% 1.95 × 10-4 5.69 × 10-4 DBahP F 0.05 % 16 17 94% 1.32 × 10-9 DBahP F 0.1 % 15 18 83% 5.27 × 10-8 1.29 × 10-7 DBaiP F 0.05 % 16 19 84% 2.58 × 10-9 DBaiP F 0.1 % 16 19 84% 2.58 × 10-9 9.81 × 10-8 DBaeF F 0.05 % 17 19 89% 3.35 × 10-9 DBaeF F 0.1 % 18 19 95% 3.05 × 10-10 1.13 × 10-917660 Cavalieri et 

al., 1977 

Complete Mice Papilloma, 

keratoacanthoma, 

carcinoma 

Acetone F 0 μmol/application 0 29 0% 

 BaP F 0.396 μmol/application 30 38 79% 4.9 × 10-12 DBahP F 0.396 μmol/application 35 39 90% 2.98 × 10-15 AA F 0.396 μmol/application 18 38 47% 3.59 × 10-6 BaA F 0.396 μmol/application 1 39 3% 0.66 
C-4 DRAFT – DO NOT CITE OR QUO

Table C-1. Dermal bioassays: dose response information for incidence data 

Record 

number Reference 

Study 

type Species 

Tumor 

type PAH Sex

Dose 

of 

PAH

Dose 

units 

Number of 

animals 

with tumors

Number 

of 

animals 

in group

% tumorbearing 

animals 

Results of 

authors' 

statistical 

analysis 

(p-value)

Fisher's 

exact 

p-value 

CochranArmitage 

trend test 

p-value Com

Initiation studies 

630 LaVoie et 

al., 1982 

Initiation Mice Primarily 

squamous cell Papilloma 

Acetone/TPA 

F 0 μg/mouse 0 20 0% 

 BaP F 30 μg/mouse 17 20 85% 1.28 × 10-8 BbF F 10 μg/mouse 9 20 45% 6.14 × 10-4 BbF F 30 μg/mouse 12 20 60% 2.25 × 10-5 BbF F 100 μg/mouse 16 20 80% 7.7 × 10-8 1.46 × 10-5 BjF F 30 μg/mouse 6 20 30% 0.01  BjF F 100 μg/mouse 11 20 55% 7.27 × 10-5 BjF F 1,000 μg/mouse 19 20 95% 1.52 × 10-10 4.67 × 10-8 BkF F 30 μg/mouse 1 20 5% 0.01  BkF F 100 μg/mouse 5 20 25% 0.02  BkF F 1,000 μg/mouse 15 20 75% 3.85 × 10-7 4.51 × 10-918570 Hecht et al., 

1974 

Initiation Mice Unspecified Acetone F 0 mg/ 0 20 0% No. surv

not repocontrolsgroup sihere 

 BaP F 0.05 mg/ 6 20 30% 0.01  CH F 1 mg/ 11 19 58% 4.51 × 10-524800 Nesnow et 

al., 1984 

Initiation Mice Papilloma Acetone M 0 nmol 0 20 0% Data at 

 Acetone F 0 nmol 1 19 5%  BaP M 200 nmol 13 18 67% <0.005  BaP F 200 nmol 10 19 53% <0.005  BlAC M 50 nmol 12 20 60% <0.005  BlAC M 100 nmol 16 17 94% <0.005  BlAC M 250 nmol 21 21 100% <0.005  BlAC M 500 nmol 16 16 100% <0.005  BlAC M 1,000 nmol 19 20 95% <0.005 
C-5 DRAFT – DO NOT CITE OR QUO

Table C-1. Dermal bioassays: dose response information for incidence data 

Record 

number Reference 

Study 

type Species 

Tumor 

type PAH Sex

Dose 

of 

PAH

Dose 

units 

Number of 

animals 

with tumors

Number 

of 

animals 

in group

% tumorbearing 

animals 

Results of 

authors' 

statistical 

analysis 

(p-value)

Fisher's 

exact 

p-value 

CochranArmitage 

trend test 

p-value Com

 BlAC F 50 nmol 13 20 65% <0.005  BlAC F 100 nmol 18 19 95% <0.005  BlAC F 250 nmol 19 21 91% <0.005  BlAC F 500 nmol 20 21 95% <0.005  BlAC F 1,000 nmol 20 20 100% <0.005  BeAC M 50 nmol 4 20 20%  BeAC M 100 nmol 4 20 20%  BeAC M 250 nmol 12 20 60% <0.005  BeAC M 500 nmol 15 20 75% <0.005  BeAC M 1,000 nmol 16 18 89% <0.005  BeAC F 50 nmol 4 20 20%  BeAC F 100 nmol 7 19 37% <0.005  BeAC F 250 nmol 10 19 53% <0.005  BeAC F 500 nmol 8 18 44% <0.005  BeAC F 1,000 nmol 18 20 90% <0.005 21420 Slaga et al., 

1980 

Initiation Mouse Papilloma Control F 0 nmoles 2 30 6% Differen

controls each cheexcept Dand BeP 

 Control F 0 μmoles 3 30 10%  Control F 0 μmoles 3 30 10%  Control F 0 nmoles 2 29 6%  Control 

POOLED 

F 0 nmoles 10 119 8% 

 BaP F 200 nmoles 20 30 67% 1.41 × 10-6 BeP F 2,000 nmoles 5 29 17% 0.33  CH F 2,000 nmoles 21 29 73% 8.38 × 10-7 DBacA F 2,000 nmoles 8 28 27% 0.07  DBahA F 100 nmoles 15 29 50% 3.52 × 10-6
C-6 DRAFT – DO NOT CITE OR QUO

Table C-1. Dermal bioassays: dose response information for incidence data 

Record 

number Reference 

Study 

type Species 

Tumor 

type PAH Sex

Dose 

of 

PAH

Dose 

units 

Number of 

animals 

with tumors

Number 

of 

animals 

in group

% tumorbearing 

animals 

Results of 

authors' 

statistical 

analysis 

(p-value)

Fisher's 

exact 

p-value 

CochranArmitage 

trend test 

p-value Com

15640 Raveh et al., 

1982 

Initiation Mice Papilloma Control F 0 μg 3 29 10% 

 BaP F 10 μg 17 29 58% 1.11 × 10-4 BaP F 25 μg 21 28 76% 5.96 × 10-7 BaP F 50 μg 24 28 87% 5.43 × 10-9 BaP F 100 μg 27 27 100% 5.50 × 10-13 BaP F 200 μg 26 26 100% 1.03 × 10-12 2.78 × 10-10 CPcdP F 10 μg 3 30 11% 0.65  CPcdP F 100 μg 11 29 39% 0.01  CPcdP F 200 μg 16 28 57% 1.90 × 10-4 2.75 × 10-6620 Hoffmann 

and Wynder 1966 

Initiation Mice Papillomas Croton oil 

control 

F 0 mg/mouse 2 30 7% 

 BaP F 0.25 mg/mouse 24 30 80% 3.80 × 10-9 DBaeF F 0.25 mg/mouse 18 30 60% 9.40 × 10-6 DBaeP F 0.25 mg/mouse 10 27 37% 0.006  DBelP F 0.25 mg/mouse 0 29 0% 0.25  DBahP F 0.25 mg/mouse 21 29 72% 1.30 × 10-7 DBaiP F 0.25 mg/mouse 12 30 40% 0.002  AA F 0.25 mg/ mouse 2 29 7% 0.68  BghiP F 0.25 mg/mouse 2 27 7% 0.65  N23eP F 0.25 mg/mouse 9 30 30% 0.02  IP F 0.25 mg/mouse 5 30 17% 0.21 13650 Cavalieri et 

al., 1981b 

Initiation Mice Papilloma Acetone/

TPA 

F 0 μmol 3 29 10% 

 BaP F 0.2 μmol 12 30 40% 0.009  CPcdP F 0.2 μmol 1 30 3% 0.29  CPcdP F 0.6 μmol 9 29 31% 0.05  CPcdP F 1.8 μmol 6 29 21% 0.24 0.14  ACEP F 0.2 μmol 0 30 0% 0.11  ACEP F 0.6 μmol 1 30 3% 0.29 
C-7 DRAFT – DO NOT CITE OR QUO

Table C-1. Dermal bioassays: dose response information for incidence data 

Record 

number Reference 

Study 

type Species 

Tumor 

type PAH Sex

Dose 

of 

PAH

Dose 

units 

Number of 

animals 

with tumors

Number 

of 

animals 

in group

% tumorbearing 

animals 

Results of 

authors' 

statistical 

analysis 

(p-value)

Fisher's 

exact 

p-value 

CochranArmitage 

trend test 

p-value Com

 ACEP F 1.8 μmol 4 30 13% 0.52 0.18 15700 Rice et al., 

1988 

Initiation Mice Unspecified Acetone F 0 μmol 1 20 5% 

 BaP F 0.1 μmol 17 19 89% <0.005  CH F 0.15 μmol 5 20 25% <0.05  CH F 0.5 μmol 18 20 90% <0.005  CH F 1.5 μmol 19 20 95% <0.005 6.39 × 10-9 CPdefC 

(4,5-MC) 

F 0.15 μmol 13 20 65% <0.005 

 CPdefC 

(4,5-MC) 

F 0.5 μmol 19 19 100% <0.005 

 CPdefC 

(4,5-MC) 

F 1.5 μmol 19 19 100% <0.005 1.90 × 10-7

 BbcAC 

(1,12-MBA) 

F 0.5 μmol 15 20 75% <0.005 

 BbcAC 

(1,12-MBA) 

F 2 μmol 18 20 90% <0.005 

 BbcAC 

(1,12-MBA) 

F 4 μmol 18 20 90% <0.005 3.03 × 10-6

1 
C-8 DRAFT – DO NOT CITE OR QUO

1 

Table C-2. Dermal bioassays: dose-response information for tumor multiplicity 

Record 

numbe

r Reference 

Study 

type Species

Tumor 

type PAH Sex

Dose of 

PAH Dose units

Number of 

animals 

with 

tumors 

Number 

of 

animals 

in 

group 

% 

Tumorbearing animals 

Results of 

authors' 

statistical 

analysis 

(p-value) 

Results of 

SRC 

statistical 

analysis 

Fisher's 

exact 

p-value

Mean 

number 

tumors/ 

animal Comm

Complete carcinogenicity 

13640 Cavalieri et al., 

1983 

Complete Mice Papilloma, 

adenoma, carcinoma 

Acetone F 0 nmol 0 29 0% 0 Number tum

animal at ricalculated 

 BaP F 2.2 nmol 2 30 7% >0.05 0.07  BaP F 6.6 nmol 2 28 7% >0.05 0.07  BaP F 20 nmol 17 30 57% <0.001 1.5  CPcdP F 22.2 nmol 2 29 7% >0.05 0.07  CPcdP F 66.6 nmol 2 29 7% >0.05 0.07  CPcdP F 200 nmol 24 29 83% <0.001 2.45 13650 Cavalieri et al., 

1981b 

Complete Mice Primarily 

squamous cell carcinoma 

Acetone US 0 μmol/ 

application

0 30 0% 0 Number tum

animal at ricalculated 

 BaP US 0.2 μmol/ 

application

30 30 100% <0.001 1.5 

 CPcdP US 0.2 μmol 

/application 

17 30 57% <0.001 0.8 

 CPcdP US 0.6 μmol/ 

application

11 30 37% <0.001 0.5 

 CPcdP US 1.8 μmol/ 

application

7 30 23% 0.0053 0.4 

 ACEP US 0.2 μmol/ 

application

0 30 0% >0.05 0 

 ACEP US 0.6 μmol/ 

application

1 30 3% >0.05 0.03 

 ACEP US 1.8 μmol/ 

application

1 30 3% >0.05 0.03 

Initiation 

630 LaVoie et al., 

1982 

Initiation Mice Primarily 

squamous cell 

papilloma 

Acetone/ TPA 

F 0 μg/mouse 0 20 0% 0 

 BaP F 30 μg/mouse 17 20 85% <0.001 4.9  BbF F 10 μg/mouse 9 20 45% <0.001 0.9  BbF F 30 μg/mouse 12 20 60% <0.001 2.3 
C-9 DRAFT – DO NOT CITE OR QUO

Table C-2. Dermal bioassays: dose-response information for tumor multiplicity 

Record 

numbe

r Reference 

Study 

type Species

Tumor 

type PAH Sex

Dose of 

PAH Dose units

Number of 

animals 

with 

tumors 

Number 

of 

animals 

in 

group 

% 

Tumorbearing animals 

Results of 

authors' 

statistical 

analysis 

(p-value) 

Results of 

SRC 

statistical 

analysis 

Fisher's 

exact 

p-value

Mean 

number 

tumors/ 

animal Comm

 BbF F 100 μg/mouse 16 20 80% <0.001 7.1  BjF F 30 μg/mouse 6 20 30% 0.01 0.6  BjF F 100 μg/mouse 11 20 55% <0.001 1.9  BjF F 1,000 μg/mouse 19 20 95% <0.001 7.2  BkF F 30 μg/mouse 1 20 5% >0.05 0.1  BkF F 100 μg/mouse 5 20 25% 0.02 0.4  BkF F 1,000 μg/mouse 15 20 75% <0.001 2.8 18570 Hecht et al., 1974 Initiation Mice Unspecifie

d 

Acetone F 0 mg/animal 0 20 0% 0 Number sur

not reported controls; ingroup size uhere. Numbtumors per at risk calcu

 BaP F 0.05 mg/animal 6 20 30% 0.01 0.5  CH F 1 mg/animal 11 19 61% <0.001 1 21420 Slaga et al., 1980 Initiation Mouse Papilloma Control F 0 nmol 2 29 6% 0.1 Different co

used for eacchemical exDBacA and 

 Control F 0 nmol 3 30 10% 0.2  Control F 0 nmol 3 30 10% 0.1  Control F 0 nmol 2 29 6% 0.1  Control 

POOLED 

F 0 nmol 10 119 8% 0.13 

 BaP F 200 nmol 20 30 67% <0.001 2.2  BeP F 2,000 nmol 5 29 17% >0.05 0.2  CH F 2,000 nmol 21 29 73% <0.001 1.6  DBacA F 2,000 nmol 8 28 27% >0.05 0.5  DBahA F 100 nmol 15 29 50% <0.001 1.4 15640 Raveh et al., 

1982 

Initiation Mice Papilloma Control F 0 μg 3 29 10% 0.2 

 BaP F 10 μg 17 29 58% <0.001 1.3  BaP F 25 μg 21 28 76% <0.001 3.8  BaP F 50 μg 24 28 87% <0.001 6.2  BaP F 100 μg 27 27 100% <0.001 8.8 
C-10 DRAFT – DO NOT CITE OR QUO

Table C-2. Dermal bioassays: dose-response information for tumor multiplicity 

Record 

numbe

r Reference 

Study 

type Species

Tumor 

type PAH Sex

Dose of 

PAH Dose units

Number of 

animals 

with 

tumors 

Number 

of 

animals 

in 

group 

% 

Tumorbearing animals 

Results of 

authors' 

statistical 

analysis 

(p-value) 

Results of 

SRC 

statistical 

analysis 

Fisher's 

exact 

p-value

Mean 

number 

tumors/ 

animal Comm

 BaP F 200 μg 26 26 100% <0.001 9  CPcdP F 10 μg 3 30 11% >0.05 0.1  CPcdP F 100 μg 11 29 39% 0.01 0.4  CPcdP F 200 μg 16 28 57% <0.001 0.9 13650 Cavalieri et al., 

1981 

Initiation Mice Papilloma Acetone/ 

TPA 

F 0 μmol 3 29 10% 0.14 

 BaP F 0.2 μmol 12 30 40% 0.009 1.2  CPcdP F 0.2 μmol 1 30 3% >0.05 0.03  CPcdP F 0.6 μmol 9 29 31% 0.05 0.31  CPcdP F 1.8 μmol 6 29 21% >0.05 0.31  ACEP F 0.2 μmol 0 30 0% >0.05 0  ACEP F 0.6 μmol 1 30 3% >0.05 0.03  ACEP F 1.8 μmol 4 30 13% >0.05 0.13 21410 Slaga et al., 1978 Initiation Mice Papillomas Acetone/ 

TPA 

F 0 μmol 2 29 6% 0.1 

 BaP F 0.2 μmol 27 29 92% <0.001 5.3  BaA F 2 μmol 17 30 57% <0.001 1.2 16310 Weyand et al., 

1992 

Initiation Mice Unspecifie

d 

 Acetone US 0 μmol 1 21 5% 0.05 

 BaP US 0.01 μmol 24 24 100% <0.01 4.08  BjF US 0.3 μmol 11 20 55% <0.01 1.75  BjF US 1 μmol 21 24 88% <0.01 4.08  BjF US 2 μmol 24 24 100% <0.01 7.17 10200 El-Bayoumy et 

al., 1982 

Initiation Mice Primarily 

squamous cell 

papilloma 

Acetone F 0 mg/mouse 1 20 5% 0.1 

 BaP F 0.05 mg/mouse 18 20 90% <0.01 7.1  CH F 1 mg/mouse 20 20 100% <0.01 7.7  Pery F 1 mg/mouse 1 20 5% 0.1  Pyr F 1 mg/mouse 4 20 20% 0.2 24300 Rice et al., 1985 Initiation Mice Unspecifie

d 

Acetone F 0 mg/mouse 2 25 8% 0.12 Mean numb

tumors/ anidigitally estfrom Figure rounded to number tum

 BaP F 0.3 mg/mouse 24 25 96% <0.001 8.04 
C-11 DRAFT – DO NOT CITE OR QUO

Table C-2. Dermal bioassays: dose-response information for tumor multiplicity 

Record 

numbe

r Reference 

Study 

type Species

Tumor 

type PAH Sex

Dose of 

PAH Dose units

Number of 

animals 

with 

tumors 

Number 

of 

animals 

in 

group 

% 

Tumorbearing animals 

Results of 

authors' 

statistical 

analysis 

(p-value) 

Results of 

SRC 

statistical 

analysis 

Fisher's 

exact 

p-value

Mean 

number 

tumors/ 

animal Comm

 CH F 1 mg/mouse 23 25 92% <0.001 5  CPdefC F 1 mg/mouse 24 24 100% <0.001 5.63 Number rep

text 

13660 Cavalieri et al., 

1991 

Initiation Mice Primarily 

papilloma 

Acetone F 0 nmol 0 24 0% 0 16 Wk expe

 BaP F 33.3 nmol 10 23 43% <0.001 0.65  BaP F 100 nmol 17 24 71% <0.001 2.75  BaP F 300 nmol 21 23 91% <0.001 5.22  DBalP F 33.3 nmol 23 24 96% <0.001 6.75  DBalP F 100 nmol 22 24 92% <0.001 7.92  DBalP F 300 nmol 24 24 100% <0.001 8.5 13660 Cavalieri et al., 

1991 

Initiation Mice Primarily 

papilloma 

Acetone F 0 nmol 0 24 0% 0 27 Wk expe

 BaP F 4 nmol 1 24 4% >0.05 0.04  BaP F 20 nmol 10 24 42% <0.001 0.75  BaP F 100 nmol 22 24 92% <0.001 3.42  DBalP F 4 nmol 22 24 92% <0.001 6.96  DBalP F 20 nmol 20 24 83% <0.001 5.29  DBalP F 100 nmol 20 24 83% <0.001 3.29 16440 Wood et al., 1980 Initiation Mice Papillomas Acetone F 0 μmol 3 30 10% 0.1 Number tum

animal at ricalculated 

 BaP F 0.1 μmol 20 30 68% <0.05 2  BaP F 0.4 μmol 22 30 73% <0.05 4.6  Pyr F 0.1 μmol 4 30 14% >0.05 0.14  Pyr F 0.4 μmol 3 30 10% >0.05 0.1  CPcdP F 0.1 μmol 3 30 10% >0.05 0.1  CPcdP F 0.4 μmol 6 30 21% >0.05 0.29 18680 Hoffmann et al., 

1972 

Initiation Mice Papillomas Acetone F 0 mg 1 30 3% 0.03 

 BaP F 0.05 mg 19 29 66% <0.001 2.3  FA F 1 mg 1 29 3% >0.05 0.03 24800 Nesnow et al., 

1984 

Initiation Mice Papillomas Acetone M 0 nmol 0 20 0% 0 

 Acetone F 0 nmol 1 19 5% 0.05  BaP M 200 nmol 12 18 67% <0.001 1.4  BaP F 200 nmol 10 19 53% 0.0015 1.5  BeAC M 50 nmol 4 20 20% >0.05 0.25 
C-12 DRAFT – DO NOT CITE OR QUO

Table C-2. Dermal bioassays: dose-response information for tumor multiplicity 

Record 

numbe

r Reference 

Study 

type Species

Tumor 

type PAH Sex

Dose of 

PAH Dose units

Number of 

animals 

with 

tumors 

Number 

of 

animals 

in 

group 

% 

Tumorbearing animals 

Results of 

authors' 

statistical 

analysis 

(p-value) 

Results of 

SRC 

statistical 

analysis 

Fisher's 

exact 

p-value

Mean 

number 

tumors/ 

animal Comm

 BeAC F 50 nmol 4 20 20% >0.05 0.25  BeAC M 100 nmol 4 20 20% >0.05 0.4  BeAC F 100 nmol 7 19 37% 0.02 0.53  BeAC M 250 nmol 12 20 60% <0.001 1.3  BeAC F 250 nmol 10 19 53% <0.001 1.1  BeAC M 500 nmol 15 20 75% <0.001 1.9  BeAC F 500 nmol 8 18 44% 0.007 1.2  BeAC M 1,000 nmol 16 18 89% <0.001 3.1  BeAC F 1,000 nmol 18 20 90% <0.001 2.2  BlAC M 50 nmol 12 20 60% <0.001 1.4  BlAC F 50 nmol 13 20 65% <0.001 1.1  BlAC M 100 nmol 16 17 94% <0.001 2.3  BlAC F 100 nmol 18 19 95% <0.001 3.1  BlAC M 250 nmol 21 21 100% <0.001 8.4  BlAC F 250 nmol 19 21 91% <0.001 4.7  BlAC M 500 nmol 16 16 100% <0.001 10.8  BlAC F 500 nmol 20 21 95% <0.001 6.6  BlAC M 1,000 nmol 19 20 95% <0.001 8.7  BlAC F 1,000 nmol 20 20 100% <0.001 10.8

1 
C-13 DRAFT – DO NOT CITE OR QUO

1 

Table C-3. Intraperitoneal bioassays: dose response information for incidence data 

SRC Statistical 

Analysis 

Record 

number Reference Species

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose

Dose 

units 

No. of animals 

with tumors 

No. of animals 

in group 

% Tumor bearing animals Results of authors' statistical analysis 

(p-value) 

Fisher's 

Exact 

p-value 

CochranArmitage 

trend test 

p-value Comme

17560 Busby et al., 

1989 

Mice Intraperitoneal Lung Adenoma + adeno- carcinoma DMSO M 0 μg (total) 13 91 0.14 Stats reported 

combined M only for each and treatmencompared to not individua

 Lung Adenoma + 

adenocarcinoma 

DMSO F 0 μg 

(total) 

7 101 0.07 

 Lung Adenoma + 

adenocarcinoma 

BaP M 59.5 μg 

(total) 

13 28 0.46 7.2 × 10-4

 Lung Adenoma + 

adenocarcinoma 

BaP F 59.5 μg 

(total) 

19 27 0.70 3.96 × 10-11

 Lung Adenoma + 

adenocarcinoma 

Pyr M 86.1 μg 

(total) 

4 23 0.17 4.60 × 10-1

 Lung Adenoma + 

adenocarcinoma 

Pyr F 86.1 μg 

(total) 

1 28 0.04 4.50 × 10-1

 Lung Adenoma + 

adenocarcinoma 

Pyr M 1,750 μg 

(total) 

2 27 0.07 2.80 × 10-1 3.13 × 10-1

 Lung Adenoma + 

adenocarcinoma 

Pyr F 1,750 μg 

(total) 

3 26 0.12 3.30 × 10-1 3.50 × 10-1

 Lung Adenoma + 

adenocarcinoma 

FA M 257.6 μg 

(total) 

5 23 0.22 2.80 × 10-4

 Lung Adenoma + 

adenocarcinoma 

FA F 257.6 μg 

(total) 

9 29 0.31 1.65 × 10-3
C-14 DRAFT – DO NOT CITE OR QUO

Table C-3. Intraperitoneal bioassays: dose response information for incidence data 

SRC Statistical 

Analysis 

Record 

number Reference Species

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose

Dose 

units 

No. of animals 

with tumors 

No. of animals 

in group 

% Tumor bearing animals Results of authors' statistical analysis 

(p-value) 

Fisher's 

Exact 

p-value 

CochranArmitage 

trend test 

p-value Comme

 Lung Adenoma + 

adenocarcinoma 

CH M 6.3 μg 

(total) 

2 27 0.07 2.80 × 10-1

 Lung Adenoma + 

adenocarcinoma 

CH F 6.3 μg 

(total) 

3 29 0.10 3.90 × 10-1

 Lung Adenoma + 

adenocarcinoma 

CH M 210 μg 

(total) 

3 20 0.15 5.85 × 10-1 8.03 × 10-1

 Lung Adenoma + 

adenocarcinoma 

CH F 210 μg 

(total) 

0 29 0.00 1.60 × 10-1 1.28 × 10-1

640 LaVoie et 

al., 1987 

Mice Intraperitoneal Lung Adenoma DMSO M 0 μmol/ mouse 0 17 0 

 Lung Adenoma DMSO F 0 μmol/

mouse 

0 18 0 

 Lung Adenoma BaP M 1.1 μmol/

mouse 

14 17 0.82 <0.005 

 Lung Adenoma BaP F 1.1 μmol/

mouse 

9 14 0.64 

 Lung Adenoma BbF M 0.5 μmol/

mouse 

2 15 0.13 >0.05 

 Lung Adenoma BbF F 0.5 μmol/

mouse 

3 17 0.18 >0.05 

 Lung Adenoma BjF M 1.1 μmol/

mouse 

11 21 0.52 <0.005 

 Lung Adenoma BjF F 1.1 μmol/

mouse 

4 18 0.22 <0.05 

 Lung Adenoma BkF M 2.1 μmol/

mouse 

1 16 0.06 >0.05 

 Lung Adenoma BkF F 2.1 μmol/

mouse 

3 18 0.17 >0.05 
C-15 DRAFT – DO NOT CITE OR QUO

Table C-3. Intraperitoneal bioassays: dose response information for incidence data 

SRC Statistical 

Analysis 

Record 

number Reference Species

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose

Dose 

units 

No. of animals 

with tumors 

No. of animals 

in group 

% Tumor bearing animals Results of authors' statistical analysis 

(p-value) 

Fisher's 

Exact 

p-value 

CochranArmitage 

trend test 

p-value Comme

 Lung Adenoma IP M 2.1 μmol/ 

mouse 

1 11 0.09 

 Lung Adenoma IP F 2.1 μmol/ 

mouse 

0 9 0 

 Liver Adenoma + 

hepatoma 

DMSO M 0 μmol/ 

mouse 

1 17 0.06 Adenoma and 

hepatoma alsreported sepa

None of anim

surviving 35 

 Liver Adenoma + 

hepatoma 

DMSO F 0 μmol/ 

mouse 

0 18 0 

 Liver Adenoma + 

hepatoma 

BaP M 1.1 μmol/ 

mouse 

13 17 0.76 <0.005 

 Liver Adenoma + 

hepatoma 

BaP F 1.1 μmol/ 

mouse 

0 14 0 

 Liver Adenoma + 

hepatoma 

BbF M 0.5 μmol/ 

mouse 

8 15 0.53 <0.005 

 Liver Adenoma + 

hepatoma 

BbF F 0.5 μmol/ 

mouse 

0 17 0 

 Liver Adenoma + 

hepatoma 

BjF M 1.1 μmol/ 

mouse 

11 21 0.52 <0.005 

 Liver Adenoma + 

hepatoma 

BjF F 1.1 μmol/ 

mouse 

0 18 0 

 Liver Adenoma + 

hepatoma 

BkF M 2.1 μmol/ 

mouse 

3 16 0.19 >0.05 

 Liver Adenoma + 

hepatoma 

BkF F 2.1 μmol/ 

mouse 

0 18 0 

 Liver Adenoma + 

hepatoma 

IP M 2.1 μmol/ 

mouse 

0 11 0 

 Liver Adenoma + 

hepatoma 

IP F 2.1 μmol/ 

mouse 

0 9 0 

 Liver or 

lung 

Adenoma + hepatoma 

DMSO M 0 μmol/ 

mouse 

1 17 0.06 
C-16 DRAFT – DO NOT CITE OR QUO

Table C-3. Intraperitoneal bioassays: dose response information for incidence data 

SRC Statistical 

Analysis 

Record 

number Reference Species

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose

Dose 

units 

No. of animals 

with tumors 

No. of animals 

in group 

% Tumor bearing animals Results of authors' statistical analysis 

(p-value) 

Fisher's 

Exact 

p-value 

CochranArmitage 

trend test 

p-value Comme

 Liver or 

lung 

Adenoma + hepatoma 

DMSO F 0 μmol/ 

mouse 

0 18 0 

 Liver or 

lung 

Adenoma + hepatoma 

BaP M 1.1 μmol/ 

mouse 

13 17 0.76 

 Liver or 

lung 

Adenoma + hepatoma 

BaP F 1.1 μmol/ 

mouse 

9 14 0.64 

 Liver or 

lung 

Adenoma + hepatoma 

BbF M 0.5 μmol/ 

mouse 

8 15 0.53 

 Liver or 

lung 

Adenoma + hepatoma 

BbF F 0.5 μmol/ 

mouse 

3 17 0.18 

 Liver or 

lung 

Adenoma + hepatoma 

BjF M 1.1 μmol/ 

mouse 

17 21 0.81 

 Liver or 

lung 

Adenoma + hepatoma 

BjF F 1.1 μmol/ 

mouse 

4 18 0.22 

 Liver or 

lung 

Adenoma + hepatoma 

BkF M 2.1 μmol/ 

mouse 

3 16 0.19 

 Liver or 

lung 

Adenoma + hepatoma 

BkF F 2.1 μmol/ 

mouse 

3 18 0.17 

 Liver or 

lung 

Adenoma + hepatoma 

IP M 2.1 μmol/ 

mouse 

1 11 0.09 

 Liver or 

lung 

Adenoma + hepatoma 

IP F 2.1 μmol/ 

mouse 

0 9 0 

7510 LaVoie et 

al., 1994 

mice intraper

itoneal 

Lung Total DMSO M 0 μmol/ 

mouse 

5 29 0.17 Surv to 1 yr 

 Lung Total DMSO F 0 μmol/ 

mouse 

4 34 0.12 

 Lung Total BaP M 1.1 μmol/ 

mouse 

24 32 0.75 <0.001 

 Lung Total BaP F 1.1 μmol/ 

mouse 

17 20 0.85 <0.001 

 Lung Total FA M 3.46 μmol/ 

mouse 

12 28 0.43 <0.05 
C-17 DRAFT – DO NOT CITE OR QUO

Table C-3. Intraperitoneal bioassays: dose response information for incidence data 

SRC Statistical 

Analysis 

Record 

number Reference Species

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose

Dose 

units 

No. of animals 

with tumors 

No. of animals 

in group 

% Tumor bearing animals Results of authors' statistical analysis 

(p-value) 

Fisher's 

Exact 

p-value 

CochranArmitage 

trend test 

p-value Comme

 Lung Total FA F 3.46 μmol/ 

mouse 

11 31 0.35 <0.05 

 Lung Total FA M 17.3 μmol/ 

mouse 

11 17 0.65 <0.005 2.84 × 10-3

 Lung Total FA F 17.3 μmol/ 

mouse 

25 29 0.86 <0.001 2.18 × 10-9

 Liver Foci + 

adenoma + carcinoma 

DMSO M 0 μmol/ 

mouse 

5 29 0.17 Foci, adenom

carcinomas areported sepa

 Liver Foci + 

adenoma + carcinoma 

DMSO F 0 μmol/ 

mouse 

2 34 0.06 

 Liver Foci + 

adenoma + carcinoma 

BaP M 1.1 μmol/ 

mouse 

27 32 0.84 <0.001 

 Liver Foci + 

adenoma + carcinoma 

BaP F 1.1 μmol/ 

mouse 

2 20 0.10 >0.05 

 Liver Foci + 

adenoma + carcinoma 

FA M 3.46 μmol/ 

mouse 

18 28 0.64 <0.001 

 Liver Foci + 

adenoma + carcinoma 

FA F 3.46 μmol/ 

mouse 

0 31 0 

 Liver Foci + 

adenoma + carcinoma 

FA M 17.3 μmol/ 

mouse 

17 17 1.00 <0.001 5.10 × 10-7

 Liver Foci + 

adenoma + carcinoma 

FA F 17.3 μmol/ 

mouse 

2 29 0.07 5.47 × 10-1

22510 Wislocki et 

al., 1986 

Mice Intraperitoneal Liver Adenoma + carcinoma DMSO M 0 nmol 2 28 0.07 Animals surv

thru weaning 
C-18 DRAFT – DO NOT CITE OR QUO

Table C-3. Intraperitoneal bioassays: dose response information for incidence data 

SRC Statistical 

Analysis 

Record 

number Reference Species

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose

Dose 

units 

No. of animals 

with tumors 

No. of animals 

in group 

% Tumor bearing animals Results of authors' statistical analysis 

(p-value) 

Fisher's 

Exact 

p-value 

CochranArmitage 

trend test 

p-value Comme

 Liver Adenoma +

carcinoma 

DMSO F 0 nmol 0 31 0 0 

 Liver Adenoma +

carcinoma 

DMSO M 0 nmol 5 45 0.11 This group st

10 wks after groups 

 Liver Adenoma +

carcinoma 

DMSO F 0 nmol 0 34 0 This group st

10 wks after groups 

 Liver Adenoma +

carcinoma 

DMSO POOLED 

M 0 nmol 7 73 0.09 

 Liver Adenoma +

carcinoma 

DMSO POOLED 

F 0 nmol 0 65 0 

 Liver Adenoma +

carcinoma 

BaP M 560 nmol 18 37 0.49 <0.05 

 Liver Adenoma +

carcinoma 

BaP F 560 nmol 0 27 0 

 Liver Adenoma +

carcinoma 

CH M 700 nmol 10 35 0.29 <0.05 This group st

10 wks after groups 

 Liver Adenoma +

carcinoma 

CH F 700 nmol 0 33 0 This group st

10 wks after groups 

 Liver Adenoma +

carcinoma 

CH M 2,800 nmol 14 34 0.41 <0.05 6 × 10-3

 Liver Adenoma +

carcinoma 

CH F 2,800 nmol 0 24 0 1 

 Liver Adenoma +

carcinoma 

BaA M 2,800 nmol 31 39 0.79 <0.05 

 Liver Adenoma +

carcinoma 

BaA F 2,800 nmol 0 32 0 
C-19 DRAFT – DO NOT CITE OR QUO

Table C-3. Intraperitoneal bioassays: dose response information for incidence data 

SRC Statistical 

Analysis 

Record 

number Reference Species

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose

Dose 

units 

No. of animals 

with tumors 

No. of animals 

in group 

% Tumor bearing animals Results of authors' statistical analysis 

(p-value) 

Fisher's 

Exact 

p-value 

CochranArmitage 

trend test 

p-value Comme

 Lung Adenoma +

carcinoma 

DMSO M 0 nmol 1 28 0.04 

 Lung Adenoma +

carcinoma 

DMSO F 0 nmol 0 31 0 

 Lung Adenoma +

carcinoma 

DMSO M 0 nmol 4 45 0.09 This group st

10 wks after groups 

 Lung Adenoma +

carcinoma 

DMSO F 0 nmol 2 34 0.06 This group st

10 wks after groups 

 Lung Adenoma +

carcinoma 

DMSO 

pooled 

M 0 nmol 5 73 0.07 

 Lung Adenoma +

carcinoma 

DMSO 

pooled 

F 0 nmol 2 65 0.03 

 Lung Adenoma +

carcinoma 

BaP M 560 nmol 13 37 0.35 <0.05 

 Lung Adenoma +

carcinoma 

BaP F 560 nmol 13 27 0.48 <0.05 

 Lung Adenoma +

carcinoma 

CH M 700 nmol 6 35 0.17 This group st

10 wks after groups 

 Lung Adenoma +

carcinoma 

CH F 700 nmol 2 33 0.06 This group st

10 wks after groups 

 Lung Adenoma +

carcinoma 

CH M 2,800 nmol 7 34 0.21 <0.05 1.1 × 10-1

 Lung Adenoma +

carcinoma 

CH F 2,800 nmol 1 24 0.04 5.6 × 10-1

 Lung Adenoma +

carcinoma 

BaA M 2,800 nmol 6 39 0.15 

 Lung Adenoma +

carcinoma 

BaA F 2,800 nmol 6 32 0.19 <0.05 
C-20 DRAFT – DO NOT CITE OR QUO

Table C-3. Intraperitoneal bioassays: dose response information for incidence data 

SRC Statistical 

Analysis 

Record 

number Reference Species

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose

Dose 

units 

No. of animals 

with tumors 

No. of animals 

in group 

% Tumor bearing animals Results of authors' statistical analysis 

(p-value) 

Fisher's 

Exact 

p-value 

CochranArmitage 

trend test 

p-value Comme

 Lymphatic 

system 

Lymphoma DMSO M 0 nmol 1 28 0.04 

 Lymphatic 

system 

Lymphoma DMSO F 0 nmol 1 31 0.03 

 Lymphatic 

system 

Lymphoma DMSO M 0 nmol 0 45 0 This group st

10 wks after groups 

 Lymphatic 

system 

Lymphoma DMSO F 0 nmol 0 34 0 This group st

10 wks after groups 

 Lymphatic 

system 

Lymphoma BaP M 560 nmol 2 37 0.05 

 Lymphatic 

system 

Lymphoma BaP F 560 nmol 4 27 0.15 

 Lymphatic 

system 

Lymphoma CH M 700 nmol 3 35 0.09 <0.05 This group st

10 wks after groups 

 Lymphatic 

system 

Lymphoma CH F 700 nmol 1 33 0.03 This group st

10 wks after groups 

 Lymphatic 

system 

Lymphoma CH M 2,800 nmol 0 34 0 2.2 × 10-1

 Lymphatic 

system 

Lymphoma CH F 2,800 nmol 0 24 0 3.9 × 10-1

 Lymphatic 

system 

Adenoma + carcinoma 

BaA M 2,800 nmol 1 39 0.03 
C-21 DRAFT – DO NOT CITE OR QUO

Table C-3. Intraperitoneal bioassays: dose response information for incidence data 

SRC Statistical 

Analysis 

Record 

number Reference Species

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose

Dose 

units 

No. of animals 

with tumors 

No. of animals 

in group 

% Tumor bearing animals Results of authors' statistical analysis 

(p-value) 

Fisher's 

Exact 

p-value 

CochranArmitage 

trend test 

p-value Comme

 Lymphatic 

system 

Adenoma + carcinoma 

BaA F 2,800 nmol 3 32 0.09 

1 
C-22 DRAFT – DO NOT CITE OR QUO

1 

Table C-4. Intraperitoneal bioassays: dose-response information for tumor multiplicity 

Record 

number Reference Species 

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose Dose units 

Number 

of animals 

with 

tumors 

Number 

of animals 

in group 

% 

Tumorbearing animals 

Results of 

authors' 

statistical 

analysis 

(p-value) 

Results of 

SRC 

statistical 

analysis 

(Fisher's 

exact 

p-value) 

Mean 

number 

tumors/ 

animal 

Std 

deviation 

of mean 

Results of 

SRC 

statistical 

analysis 

(t-test 

p-value) C

17560 Busby et al., 1989 Mice Intraperitoneal Lung Adenoma+ adenocarcinoma DMSO M 0 μg (total) 13 91 0.14 0.15 0.38 S

re

ca

 Lung Adenoma+ 

adenocarcinoma 

DMSO F 0 μg (total) 7 101 0.07 0.08 0.30 

 Lung Adenoma+ 

adenocarcinoma 

BaP M 59.5 μg (total) 13 28 0.46 <0.001 0.71 1.01 <0.001 

 Lung Adenoma+ 

adenocarcinoma 

BaP F 59.5 μg (total) 19 27 0.70 <0.001 1.19 1.09 <0.001 

 Lung Adenoma+ 

adenocarcinoma 

Pyr M 86.1 μg (total) 4 23 0.17 >0.05 0.17 0.38 >0.05 

 Lung Adenoma+ 

adenocarcinoma 

Pyr F 86.1 μg (total) 1 28 0.04 >0.05 0.04 0.21 >0.05 

 Lung Adenoma+ 

adenocarcinoma 

Pyr M 1,750 μg (total) 2 27 0.07 >0.05 0.07 0.26 >0.05 

 Lung Adenoma+ 

adenocarcinoma 

Pyr F 1,750 μg (total) 3 26 0.12 >0.05 0.12 0.31 >0.05 

 Lung Adenoma+ 

adenocarcinoma 

FA M 257.6 μg (total) 5 23 0.22 >0.05 0.22 0.43 >0.05 

 Lung Adenoma+ 

adenocarcinoma 

FA F 257.6 μg (total) 9 29 0.31 0.00165 0.41 0.70 <0.0001 

 Lung Adenoma+ 

adenocarcinoma 

CH M 6.3 μg (total) 2 27 0.07 >0.05 0.07 0.26 >0.05 

 Lung Adenoma+ 

adenocarcinoma 

CH F 6.3 μg (total) 3 29 0.10 >0.05 0.1 0.32 >0.05

 Lung Adenoma+ 

adenocarcinoma 

CH M 210 μg (total) 3 20 0.15 >0.05 0.15 0.36 >0.05 

 Lung Adenoma+ 

adenocarcinoma 

CH F 210 μg (total) 0 29 0.00 >0.05 0 0.00 >0.05 

7510 LaVoie et al., 1994 Mice intraperitoneal Lung Total DMSO M 0 μmol/mouse 5 29 0.17 0.17 S

1 

 Lung Total DMSO F 0 μmol/mouse 4 34 0.12 0.15 Lung Total BaP M 1.1 μmol/mouse 24 32 0.75 <0.001 4.3  Lung Total BaP F 1.1 μmol/mouse 17 20 0.85 <0.001 3.55  Lung Total FA M 3.46 μmol/mouse 12 28 0.43 <0.05 0.64 
C-23 DRAFT – DO NOT CITE OR QUO

Table C-4. Intraperitoneal bioassays: dose-response information for tumor multiplicity 

Record 

number Reference Species 

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose Dose units 

Number 

of animals 

with 

tumors 

Number 

of animals 

in group 

% 

Tumorbearing animals 

Results of 

authors' 

statistical 

analysis 

(p-value) 

Results of 

SRC 

statistical 

analysis 

(Fisher's 

exact 

p-value) 

Mean 

number 

tumors/ 

animal 

Std 

deviation 

of mean 

Results of 

SRC 

statistical 

analysis 

(t-test 

p-value) C

 Lung Total FA F 3.46 μmol/mouse 11 31 0.35 <0.05 0.35  Lung Total FA M 17.3 μmol/mouse 11 17 0.65 <0.005 1.12  Lung Total FA F 17.3 μmol/mouse 25 29 0.86 <0.001 2.45  Liver Foci + 

adenoma + carcinoma 

DMSO M 0 μmol/mouse 5 29 0.17 0.41 

 Liver Foci + 

adenoma + carcinoma 

DMSO F 0 μmol/mouse 2 34 0.06 0.06 T

ca

bp

 Liver Foci + 

adenoma + carcinoma 

BaP M 1.1 μmol/mouse 27 32 0.84 <0.001 4.53 

 Liver Foci + 

adenoma + carcinoma 

BaP F 1.1 μmol/mouse 2 20 0.10 >0.05 0.3 

 Liver Foci + 

adenoma + carcinoma 

FA M 3.46 μmol/mouse 18 28 0.64 <0.001 1.86 

 Liver Foci + 

adenoma + carcinoma 

FA F 3.46 μmol/mouse 0 31 0 0 

 Liver Foci + 

adenoma + carcinoma 

FA M 17.3 μmol/mouse 17 17 1.00 <0.001 7.53 

 Liver Foci + 

adenoma + carcinoma 

FA F 17.3 μmol/mouse 2 29 0.07 0.07 

22510 Wislocki et al., 1986 Mice intraperitoneal Liver Adenoma + carcinoma DMSO M 0 nmol 2 28 0.07 0.07 A

suthw

 Liver Adenoma + 

carcinoma 

DMSO F 0 nmol 0 31 0 0 

 Liver Adenoma + 

carcinoma 

DMSO M 0 nmol 5 45 0.11 0.11 T

st1o

 Liver Adenoma + 

carcinoma 

DMSO F 0 nmol 0 34 0 0 T

st1o

 Liver Adenoma + 

carcinoma 

DMSO POOLED 

M 0 nmol 7 73 0.09 0.096 

 Liver Adenoma + 

carcinoma 

DMSO POOLED 

F 0 nmol 0 65 0 0 
C-24 DRAFT – DO NOT CITE OR QUO

Table C-4. Intraperitoneal bioassays: dose-response information for tumor multiplicity 

Record 

number Reference Species 

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose Dose units 

Number 

of animals 

with 

tumors 

Number 

of animals 

in group 

% 

Tumorbearing animals 

Results of 

authors' 

statistical 

analysis 

(p-value) 

Results of 

SRC 

statistical 

analysis 

(Fisher's 

exact 

p-value) 

Mean 

number 

tumors/ 

animal 

Std 

deviation 

of mean 

Results of 

SRC 

statistical 

analysis 

(t-test 

p-value) C

 Liver Adenoma + 

carcinoma 

BaP M 560 nmol 18 37 0.49 <0.05 1.46 

 Liver Adenoma + 

carcinoma 

BaP F 560 nmol 0 27 0 >0.05 0 

 Liver Adenoma + 

carcinoma 

Pyr M 200 nmol 0 29 0 >0.05 0 

 Liver Adenoma + 

carcinoma 

Pyr F 200 nmol 0 31 0 >0.05 0 

 Liver Adenoma + 

carcinoma 

Pyr M 700 nmol 3 25 0.12 >0.05 0.12 T

st1o

 Liver Adenoma + 

carcinoma 

Pyr F 700 nmol 0 49 0 >0.05 0 T

st1o

 Liver Adenoma + 

carcinoma 

Pyr M 2,800 nmol 3 14 0.21 >0.05 0.21 

 Liver Adenoma + 

carcinoma 

Pyr F 2,800 nmol 0 18 0 >0.05 0 

 Liver Adenoma + 

carcinoma 

CH M 700 nmol 10 35 0.29 <0.05 0.86 T

st1o

 Liver Adenoma + 

carcinoma 

CH F 700 nmol 0 33 0 >0.05 0 T

st1o

 Liver Adenoma + 

carcinoma 

CH M 2,800 nmol 14 34 0.41 <0.05 1.03 

 Liver Adenoma + 

carcinoma 

CH F 2,800 nmol 0 24 0 >0.05 0 

 Liver Adenoma + 

carcinoma 

BaA M 2,800 nmol 31 39 0.79 <0.05 2.38 

 Liver Adenoma + 

carcinoma 

BaA F 2,800 nmol 0 32 0 >0.05 0 

13610 Busby et al., 1984 mice intraperitoneal Lung Adenoma + carcinoma DMSO M 0 mg (total) 1 27 0.04 0.04 0.21  Lung Adenoma + 

carcinoma 

DMSO F 0 mg (total) 4 28 0.14 0.14 0.37 

 Lung Adenoma + 

carcinoma 

BaP M 0.28 mg (total) 24 25 0.96 <0.001 4.32 3.5 <0.001 

 Lung Adenoma + 

carcinoma 

BaP F 0.28 mg (total) 25 27 0.93 <0.001 3.7 3.10 <0.001 

 Lung Adenoma + 

carcinoma 

BaP M 1.4 mg (total) 16 20 0.80 <0.001 10.15 13.0 <0.001 N

 Lung Adenoma + 

carcinoma 

BaP F 1.4 mg (total) 21 24 0.88 <0.001 4.25 4.70 <0.001 N

 Lung Adenoma + 

carcinoma 

FA M 0.7 mg (total) 7 31 0.23 0.0412 0.29 0.84 >0.05 
C-25 DRAFT – DO NOT CITE OR QUO

Table C-4. Intraperitoneal bioassays: dose-response information for tumor multiplicity 

Record 

number Reference Species 

Exposure 

route 

Target 

organ Tumor type PAH Sex Dose Dose units 

Number 

of animals 

with 

tumors 

Number 

of animals 

in group 

% 

Tumorbearing animals 

Results of 

authors' 

statistical 

analysis 

(p-value) 

Results of 

SRC 

statistical 

analysis 

(Fisher's 

exact 

p-value) 

Mean 

number 

tumors/ 

animal 

Std 

deviation 

of mean 

Results of 

SRC 

statistical 

analysis 

(t-test 

p-value) C

 Lung Adenoma + 

carcinoma 

FA F 0.7 mg (total) 3 20 0.15 >0.05 0.15 0.49 >0.05 

 Lung Adenoma + 

carcinoma 

FA M 3.5 mg (total) 20 27 0.74 <0.001 1.52 1.66 <0.001 N

v

 Lung Adenoma + 

carcinoma 

FA F 3.5 mg (total) 8 21 0.38 >0.05 0.52 0.82 0.0343 N

innv

24590 Nesnow et al., 1998b mice intraperitoneal Lung NS Control M 0 mg/kg 6 20 0.30 0.53 0.72 P

cfr

pN

 Lung NS BaP M 5 mg/kg 6 20 0.30 >0.05 0.45 0.80 >0.05  Lung NS BaP M 10 mg/kg 7 17 0.41 >0.05 0.53 0.78 >0.05  Lung NS BaP M 50 mg/kg 19 19 1.00 <0.001 4.37 2.74 <0.001  Lung NS BaP M 100 mg/kg 16 16 1.00 0.0018 12.75 4.28 <0.001  Lung NS BaP M 200 mg/kg 24 24 1.00 <0.001 32.96 10.23 <0.001  Lung NS BbF M 10 mg/kg 9 18 0.50 >0.05 0.67 0.75 >0.05  Lung NS BbF M 50 mg/kg 16 20 0.80 >0.05 2.00 1.82 0.0022 N

in

 Lung NS BbF M 100 mg/kg 20 20 1.00 <0.001 5.30 3.21 <0.001  Lung NS BbF M 200 mg/kg 19 19 1.00 <0.001 6.95 3.52 <0.001  Lung NS CPcdP M 10 mg/kg 8 20 0.40 >0.05 0.55 0.80 >0.05  Lung NS CPcdP M 50 mg/kg 20 20 1.00 <0.001 4.75 2.12 <0.001  Lung NS CPcdP M 100 mg/kg 19 19 1.00 <0.001 32.21 15.15 <0.001  Lung NS CPcdP M 200 mg/kg 19 19 1.00 <0.001 97.68 28.68 <0.001  Lung NS DBahA M 1.25 mg/kg 12 18 0.67 >0.05 1.44 1.46 0.0229 N

in

 Lung NS DBahA M 2.5 mg/kg 18 19 0.95 0.0053 3.05 1.90 <0.001  Lung NS DBahA M 5 mg/kg 20 20 1.00 <0.001 13.05 5.99 <0.001  Lung NS DBahA M 10 mg/kg 19 19 1.00 <0.001 32.16 10.78 <0.001 24590 Nesnow et al., 1998b mice intraperitoneal Lung NS Control M 0 mg/kg 15 30 0.50 0.67 0.80  Lung NS DBalP M 0.3 mg/kg 13 33 0.39 >0.05 0.42 0.56 >0.05  Lung NS DBalP M 1.5 mg/kg 33 34 0.97 <0.001 4.32 2.86 <0.001  Lung NS DBalP M 3 mg/kg 35 35 1.00 <0.001 7.49 3.79 <0.001  Lung NS DBalP M 6 mg/kg 30 30 1.00 <0.001 16.10 7.26 <0.001 11190 Mass et al., 1993 mice intraperitoneal Lung NS Control US 0 mg/kg 19 34 0.56 0.85 0.9  NS BaP US 20 mg/kg 10 16 0.63 >0.05 1 1 >0.05 NS BaP US 50 mg/kg 15 16 0.94 0.0065 3.9 2.9 <0.001  NS BaP US 100 mg/kg 14 14 1.00 0.0017 5.9 3.3 <0.001  NS BjAC US 20 mg/kg 12 12 1.00 0.0036 60.3 14.6 <0.001  NS BjAC US 50 mg/kg 13 13 1.00 0.0025 140.6 21.5 <0.001  NS BjAC US 100 mg/kg 14 14 1.00 0.0017 97.6 28.2 <0.001 

1 2 
C-26 DRAFT – DO NOT CITE OR QUO

Table C-5. Lung implantation bioassays: dose response information for incidence data 

SRC statistical analysis 

Record 

number Reference Species 

Target 

organ Tumor type PAH Dose 

Dose 

units 

Number

of 

animals 

with 

tumors

Number

of 

animals 

in group

% Tumorbearing 

animals 

Fisher's 

exact 

p-value 

CochranArmitage trend 

test 

p-value Comments 

17940 Deutsch-Wenzel et 

al., 1983 

Rat Lung Epidermoid 

carcinoma 

Untreated control 

0 mg 0 35 0.00 

 Vehicle 

control 

0 mg 0 35 0.00 

 BaP 0.1 mg 4 35 0.11 5.70 × 10-2 BaP 0.3 mg 21 35 0.60 6.02 × 10-9 BaP 1 mg 33 35 0.94 5.93 × 10-18 1.57 × 10-17 BbF 0.1 mg 0 35 0.00  BbF 0.3 mg 1 35 0.03 5 × 10-1 BbF 1 mg 9 35 0.26 1 × 10-3 5.12 × 10-7 BeP 0.2 mg 0 35 0.00  BeP 1 mg 0 30 0.00  BeP 5 mg 1 35 0.03 5 × 10-1 9.49 × 10-2 BjF 0.2 mg 1 35 0.03 5 × 10-1 BjF 1 mg 3 35 0.09 1.2 × 10-1 BjF 5 mg 18 35 0.51 1.96 × 10-7 1.28 × 10-11 BkF 0.16 mg 0 35 0.00  BkF 0.83 mg 3 31 0.10 1 × 10-1 BkF 4.15 mg 12 27 0.44 8.05 × 10-6 1.03 × 10-9 IP 0.16 mg 3 35 0.09 1.20 × 10-1 IP 0.83 mg 8 35 0.23 2 × 10-3 IP 4.15 mg 21 35 0.60 6.02 × 10-9 2.09 × 10-10 AA 0.16 mg 1 35 0.03 5 × 10-1 AA 0.83 mg 19 35 0.54 6.4 × 10-8 1.13 × 10-10 BghiP 0.16 mg 0 35 0.00  BghiP 0.83 mg 1 35 0.03 1.2 × 10-1 BghiP 4.15 mg 4 34 0.12 5.4 × 10-2 2.47 × 10-3 Lung Pleomorphic 

sarcoma 

Untreated control 

0 mg 0 35 0.00 

 Vehicle 

control 

0 mg 0 35 0.00 
C-27 DRAFT – DO NOT CITE OR QUO

Table C-5. Lung implantation bioassays: dose response information for incidence data 

SRC statistical analysis 

Record 

number Reference Species 

Target 

organ Tumor type PAH Dose 

Dose 

units 

Number

of 

animals 

with 

tumors

Number

of 

animals 

in group

% Tumorbearing 

animals 

Fisher's 

exact 

p-value 

CochranArmitage trend 

test 

p-value Comments 

 BaP 0.1 mg 6 35 0.17 1.2 × 10-2 BaP 0.3 mg 2 35 0.06 2.5 × 10-1 BaP 1 mg 0 35 0.00 1.36 × 10-1 BbF 0.1 mg 1 35 0.03 1.2 × 10-1 BbF 0.3 mg 2 35 0.06 2.5 × 10-1 BbF 1 mg 4 35 0.11 6. × 10-2 7.55 × 10-3 BeP 0.2 mg 0 35 0.00  BeP 1 mg 1 30 0.03  BeP 5 mg 0 35 0.00  BjF 0.2 mg 0 35 0.00  BjF 1 mg 0 35 0.00  BjF 5 mg 0 35 0.00  BkF 0.16 mg 0 35 0.00  BkF 0.83 mg 0 31 0.00  BkF 4.15 mg 0 27 0.00  IP 0.16 mg 1 35 0.03 1.2 × 10-1 IP 0.83 mg 0 35 0.00  IP 4.15 mg 0 35 0.00  AA 0.16 mg 0 35 0.00  AA 0.83 mg 0 35 0.00  BghiP 0.16 mg 0 35 0.00  BghiP 0.83 mg 0 35 0.00  BghiP 4.15 mg 0 34 0.00  Lung Carcinoma+ 

sarcoma 

Untreated control 

0 mg 0 35 0.00 

 Vehicle 

control 

0 mg 0 35 0.00 

 BaP 0.1 mg 10 35 0.29 4.63 × 10-4 BaP 0.3 mg 23 35 0.66 4.7 × 10-10 BaP 1 mg 33 35 0.94 5.9 × 10-19 3.66 × 10-9
C-28 DRAFT – DO NOT CITE OR QUO

Table C-5. Lung implantation bioassays: dose response information for incidence data 

SRC statistical analysis 

Record 

number Reference Species 

Target 

organ Tumor type PAH Dose 

Dose 

units 

Number

of 

animals 

with 

tumors

Number

of 

animals 

in group

% Tumorbearing 

animals 

Fisher's 

exact 

p-value 

CochranArmitage trend 

test 

p-value Comments 

 BbF 0.1 mg 1 35 0.03 1.2 × 10-1 BbF 0.3 mg 3 35 0.09 1.2 × 10-1 BbF 1 mg 13 35 0.37 3.1 × 10-5 9.63 × 10-8 BeP 0.2 mg 0 35 0.00  BeP 1 mg 1 30 0.03  BeP 5 mg 1 35 0.03 1.2 × 10-1 3.23 × 10-1 BjF 0.2 mg 1 35 0.03 1.2 × 10-1 BjF 1 mg 3 35 0.09 1.20 × 10-1 BjF 5 mg 18 35 0.51 1.96 × 10-7 1.28 × 10-11 BkF 0.16 mg 0 35 0.00  BkF 0.83 mg 3 31 0.10 1 × 10-1 BkF 4.15 mg 12 27 0.44 8.05 × 10-4 1.03 × 10-9 IP 0.16 mg 4 35 0.11 6 × 10-2 IP 0.83 mg 8 35 0.23 2 × 10-3 IP 4.15 mg 21 35 0.60 6.02 × 10-9 7.56 × 10-10 AA 0.16 mg 1 35 0.03  AA 0.83 mg 19 35 0.54 6.4 × 10-8 1.13 × 10-10 BghiP 0.16 mg 0 35 0.00  BghiP 0.83 mg 1 35 0.03  BghiP 4.15 mg 4 34 0.12 5.4 × 10-2 2.47 × 10-322000 Wenzel-Hartung et 

al., 1990 

Rat Lung Carcinoma Untreated 

control 

0 mg/

animal 

0 35 0.00 ED10, relative 

potencies repor

 Vehicle 

control 

0 mg/

animal 

0 35 0.00 

 BaP 0.03 mg/

animal 

3 35 0.09 1.2 × 10-1

 BaP 0.1 mg/

animal 

11 35 0.31 1.93 × 10-4

 BaP 0.3 mg/

animal 

27 35 0.77 1.29E × 10-12 8.85 × 10-15
C-29 DRAFT – DO NOT CITE OR QUO

Table C-5. Lung implantation bioassays: dose response information for incidence data 

SRC statistical analysis 

Record 

number Reference Species 

Target 

organ Tumor type PAH Dose 

Dose 

units 

Number

of 

animals 

with 

tumors

Number

of 

animals 

in group

% Tumorbearing 

animals 

Fisher's 

exact 

p-value 

CochranArmitage trend 

test 

p-value Comments 

 PH 1 mg/

animal 

0 35 0.00 

 PH 3 mg/

animal 

0 35 0.00 

 PH 10 mg/

animal 

1 35 0.03 5 × 10-1 1 

 CH 1 mg/

animal 

5 35 0.14 2.7 × 10-2

 CH 3 mg/

animal 

10 35 0.29 4.63 × 10-4 7.96 × 10-4

 DBahA 0.1 mg/

animal 

20 35 0.57 2.01 × 10-8

1 
C-30 DRAFT – DO NOT CITE OR QUOTE 

Table C-6. In vitro bacterial mutagenicity: data use 

Record 

number Reference

Data 

source Data points Basis for RPF approach Comments 

17030 Andrews et 

al., 1978 

Figure 1 Dose (μg) and # of revertant 

colonies for DBacA, DBajA, 

DBahA, AA, BghiP, BeP, BaP 

Point estimate TA100 with Ar S9 

23830 Baker et 

al., 1980 

Table 2 Use data for guinea pig-MC S9 

only (column D); dose in 

μg/plate and # of revertant 

colonies; BaP, DBaiP, BaA, 

DBacA, DBahA 

Point estimate Table 2 TA100 with guinea 

pig-MC S9; Table 1 

data not used, 

different S9 mix 

used for each of 

three experiments 

23660 Bartsch et 

al., 1980 

Appendix 

table 

Use data for BaA and BaP; 

dose in μmol/plate and 

mutagenic activity in 

revertants/μmol. 

Point estimate. TA100 rat MC S9 

17380 Bos et al., 

1988 

Table 1 Use TA100 strain only; dose 

(μg/plate) and # of revertant 

colonies/plate for PH, Pyr, BaP 

Derive point estimate for 

BaP (use PH control as 

background); continuous 

model PH and Pyr using the 

BaP response as the BMR 

TA100 with rat Ar 

S9 

17590 Carver et 

al., 1986 

Figure 1 Use curves for BaP, BaA, 

BghiF, and Pery; use 400 μL 

S9 per plate (last data point on 

x-axis); each curve is different 

dose in μg/plate, use hamster 

data; revertants per plate is yaxis 

Point estimate; use highest 

dose in hamster, except for 

perylene (use 10 μg/plate); 

this is maximal response in 

hamsters 

TA100 with hamster 

Ar S9; multi-dose 

data but not SD was 

reported 

17630 Cavalieri et 

al., 1981a 

Figure 1 DR curves for BaP, CPcdP 

(CPEP in fig.), and ACEP 

(CPAP in fig.); dose as μM, 

response as mutant fraction x 

105 

Model as quantal data 

(mutant fraction reported) 

TM677 with Ar S9 

9620 Chang et 

al., 2002 

Figure 7 DR curves for BghiF, BcPH, 

and BaP; dose (μg/plate) and 

revertants/plate 

Point estimate; use 

5 μg/plate dose for BghiF 

and BaP; use 10 μg/plate for 

BcPH 

TA100 with rat Ar 

S9; SD not available 

from graph (reported 

for some data points, 

but not all) 

24030 De Flora et 

al., 1984 

Table 2 Table provides potency 

estimates as revertants/nmol 

for BaA, Pery, BaP and BeP 

Calculate the RPF ratio 

using the potency estimates 

provided 

Determine strain 

used to calculate 

potencies; rat Ar S9 

18050 Eisenstadt 

and Gold, 

1978 

Figure 2B Use TA100 data for BaP and 

CPcdP (open circles); dose is 

1 μg for CPcdP and 2 μg for 

BaP (legend); use the same S9 

concentration (20 μL/plate) 

Point estimate; single point 

data (20 μL S9/plate) 

TA100 with rat Ar 

S9; μL S9 that 

maximizes the BaP 

response does not 

produce maximal 

response for CPcdP 

18180 Florin et 

al., 1980 

Table III Use TA100 data for BaA, CH, 

and BaP, use TA98 data for 

Pery, CO, and BaP; dose is 

indicated as optimal dose 

(μmoles/plate) and # 

revertants/ plate 

Point estimate; please note 

that reported response 

includes subtraction of 

spontaneous revertants 

(control); need to use 

formula for added risk; make 

sure to flag in comments 

Note that data for 

both TA100 and 

TA98 strains were 

used; BaP results 

were provided for 

each; rat MC S9 
C-31 DRAFT – DO NOT CITE OR QUOTE 

Table C-6. In vitro bacterial mutagenicity: data use 

Record 

number Reference

Data 

source Data points Basis for RPF approach Comments 

24080 Gibson et 

al., 1978 

Table 1 

(BaP) 

Table 3 

(PAHs) 

Use data for TA98; in Table 1 

use Expt. No.1 for BaP; in 

Table 3 use data for DBahA, 

Tphen, BaA, BghiP, CH, FE, 

Pyr; dose as μg/plate, response 

as increase in revertants 

Point estimate; use the dose 

associated with the maximum response (if reported 

as a range, do not use); 

controls were reported as 

negative (no mutagenic or 

toxic response) 

TA98 with nonenzymatic induction 

(gamma irradiation); 

multi-dose data but 

not SD was reported

14080 Gold and 

Eisenstadt, 

1980 

Table 2 Use data for 3-MC induction at 

50 μL S9/plate; dose is 4 nmol 

for BaP and CPcdP, results as 

revertants/plate 

Point estimate TA100 using 50 μL 

of rat MC S9; 

important to note 

that maximal 

response for CPcdP 

occurred at much 

lower dose of S9 

(5 μL/plate) 

18650 Hermann, 

1981 

Table 1 Table provides potency 

estimates as revertants/nmol 

for BbA, BaA, CH, FA, 

Tphen, BeP, DBacA, DBahA, 

BbF, Pery, DBalP, DBaiP, AA, 

CO; potency of BaP in legend 

as 100 revertants/nmol 

Calculate the RPF ratio 

using the potency estimates 

provided 

TA98 with rat Ar 

S9; potency 

estimates were 

calculated from the 

linear portion of the 

DR curve 

10670 Johnsen et 

al., 1997 

Figure 2 Use data for PCB microsomes 

for BaP, BjAC, BlAC; dose as 

μg/plate, response as revertants

Model to derive EDsd1; 

need to extract SDs from 

graph; control response is 

113 ± 9 revertants per plate 

(see legend); add control 

response to each response 

for modeling (it was 

subtracted prior to graphing)

TA98 with PCB 

microsomes 

19000 Kaden et 

al., 1979 

Table 1 RPFs calculated for AN, ANL, 

Pyr, BbFE, CPcdP, BaA, CH, 

Tphen, FA, BeP, Pery, BghiP, 

AA, DBacA, DBahA, DBbeF 

NA TM677 with Ar S9 

and PB S9 

24680 Lafleur et 

al., 1993 

Figures 

3 and 4 

Use DR curves for BaP, 

BghiF, CPcdP, CPhiACEA 

(CPAA), ACEA (AA), 

CPhiAPA (CPAP), APA (AP); 

dose as μg/mL, response as 

mutant fraction (×105

) 

Model as quantal data 

(mutant fraction reported) 

Forward mutation to 

8-azaguanine 

resistance in TM677 

with rat AR S9 

19320 LaVoie et 

al., 1979 

Table VI Use data for TA98 for BaP, 

BeP, and Pery; 10 μg dose and 

response as revertants/plate 

Point estimate; use 20 μg for 

BaP; 10 μg for BeP; and 

20 μg for Pery 

TA98 with rat Ar 

S9; for BeP and 

Pery the maximal 

response was in 

TA100 

23650 McCann et 

al., 1975 

Table 1 Table provides potency 

estimates as revertants/nmol 

for DBaiP, BaP, BeP, DBacA, 

DBahA, CH, BaA 

Calculate the RPF ratio 

using the potency estimates 

provided 

Multiple strains, rat 

Ar S9 
C-32 DRAFT – DO NOT CITE OR QUOTE 

Table C-6. In vitro bacterial mutagenicity: data use 

Record 

number Reference

Data 

source Data points Basis for RPF approach Comments 

20220 Pahlman 

and 

Pelkonen, 

1987 

Table 1 Use data for rat-MC induced 

(last column); potency 

estimates are provided as 

revertants/nmol for BaA, CH, 

Tphen, DBacA, DBahA 

Calculate the RPF ratio 

using the potency estimates 

provided 

TA100 with rat MC 

S9 

20450 Phillipson 

and 

Ioannides, 

1989 

Figures 

2 and 3 

Use the curve for hamster S9 

(open triangles); data for BaP, 

DBaiP, BaA, and DBahA, dose 

as μg/plate, revertants/plate 

Point estimate; use 

10 μg/plate for BaP, 

DBahA; 20 μg/plate BaA, 

DBaiP 

TA100 with hamster 

S9; multi-dose data 

but not SD was 

reported 

21000 Sakai et 

al., 1985 

Table 3 Use data for TA97 +S9 for FE, 

AC, PH, FA, Ch, Pyr, BaP, 

BeP, Pery, BghiP, CO; dose 

μg, response as revertants per 

plate 

Point estimate; use 10 μg for 

AC, PH, FA, BaP, BeP; use 

5 μg for FE; use 20 μg for 

CH, Pyr, BghiP; use 4 μg for 

Pery; use 100 μg for CO 

TA97 with rat Ar 

S9; multi-dose data 

but not SD was 

reported 

11860 Sangaiah et 

al., 1983 

Figure 2 Use data for BjAC and BaP; 

dose as μg/plate, response as 

revertants/plate 

Point estimate; use 

10 μg/plate for BjAC; use 

6 μg/plate for BaP 

TA98 with rat Ar 

S9; multi-dose data 

but not SD was 

reported 

21360 Simmon, 

1979a 

Table 1 Use data for TA100 for BaA, 

BaP, BeP; dose as μg, response 

as revertants/plate after 

subtracting background 

Point estimate TA100 with rat Ar 

S9 

21640 Teranishi 

et al., 1975

Table I 

and 

Figure 3 

Use data for TA1538 for 

DBaiP and BaP; use data in 

Figure 3 for TA 1538, PB and 

DBahA-induced S9 (open 

circles) for DBaeP 

Point estimate TA1538 with rat PB 

S9 for DBaiP; 

TA1538 with PB 

and DBahA S9 for 

DBaeP 

16180 Utesch et 

al., 1987 

Figures 

2 and 3 

Use data for homogenized 

hepatocytes (open circles) for 

BaA and BaP; dose as 

μg/plate, response as 

revertants/plates 

Point estimate; use 

12.5 μg/plate for BaP; use 

25 μg/plate for BaA 

TA100 with homogenized hepatocytes 

from Ar treated rats; 

multi-dose data but 

not SD was reported

16440 Wood et 

al., 1980 

Chart 3A Use DR curves for BaP and 

CPcdP; dose as nmol, response 

as revertants/plate 

Point estimate; use 15 nmol 

for BaP and CPcdP 

TA98 with purified 

microsomal P450; 

multi-dose data but 

not SD was reported
C-33 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

17030 Andrews et al., 

1978 

TA100 ArS9 Control 0 μg 150 Revertant 

colonies 

 BaP 250 μg 1,681 Revertant 

colonies 

 DBacA 10 μg 2,957 Revertant 

colonies 

 DBajA 10 μg 843 Revertant 

colonies 

 DBahA 25 μg 617 Revertant 

colonies 

 AA 250 μg 1,796 Revertant 

colonies 

 BghiP 100 μg 793 Revertant 

colonies 

 BeP 1,000 μg 643 Revertant 

colonies 

23830 Baker et al., 1980 TA100 Guinea pigMC Control 0 μg/plate 134 Revertant colonies 18  BaP 2.5 μg/plate 1,278 Revertant 

colonies 

10 97 

 DBaiP 5 μg/plate 737 Revertant 

colonies 

10 73 

 BaA 10 μg/plate 947 Revertant 

colonies 

10 47 

 DBacA 2.5 μg/plate 1,738 Revertant 

colonies 

10 88 

 DBahA 5 μg/plate 1,331 Revertant 

colonies 

10 98 

23660 Bartsch et al., 1980 TA100 Rat MC S9 BaP 0.027 μmol/plate 29,000 Revertants/

plate 

 Control respon

subtracted. 

 BaA 0.067 μmol/plate 6,000 Revertants/

plate 

 Control respon

subtracted. 

17380 Bos et al., 1988 TA100 Rat ArS9 BaP 7.5 μg/plate 824 Revertants/

plate 

3 Replicates 21 12 

 Control 0 μg/plate 85 Revertants/

plate 

3 Replicates 12 7 
C-34 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 PH 1 μg/plate 108 Revertants/

plate 

3 Replicates 10 6 

 PH 5 μg/plate 167 Revertants/

plate 

3 Replicates 5 3 

 PH 25 μg/plate 240 Revertants/

plate 

3 Replicates 10 6 

 Control 0 μg/plate 86 Revertants/

plate 

3 Replicates 7 4 

 Pyr 1 μg/plate 93 Revertants/

plate 

3 Replicates 9 5 

 Pyr 5 μg/plate 164 Revertants/

plate 

3 Replicates 23 13 

 Pyr 25 μg/plate 279 Revertants/

plate 

3 Replicates 10 6 

17590 Carver et al., 1986 TA100 Hamster 

ArS9 

Control 0 μg/plate 140 Revertants/

plate 

 Control curves 

difficult to digitize; controvalue estimated from BaP graph and used for all

 BaP 1 μg/plate 141 Revertants/

plate 

 Continuous dat

no SD 

 BaP 10 μg/plate 482 Revertants/

plate 

 BaP 50 μg/plate 1,035 Revertants/

plate 

 BaA 15 μg/plate 346 Revertants/

plate 

 BaA 40 μg/plate 892 Revertants/

plate 

 BaA 50 μg/plate 1,263 Revertants/

plate 

 BghiF 10 μg/plate 333 Revertants/

plate 

 BghiF 25 μg/plate 727 Revertants/

plate 
C-35 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 BghiF 50 μg/plate 985 Revertants/

plate 

 Perylene 5 μg/plate 195 Revertants/

plate 

 Perylene 10 μg/plate 993 Revertants/

plate 

 Perylene 15 μg/plate 922 Revertants/

plate 

17630 Cavalieri et al., 

1981a 

TM677 Ar S9 Control 0 μM 5 Mutants 1 × 10

5 Survivors 0.000050 Control value estimated 

 BaP 10 μM 15 Mutants 1 × 10

5 Survivors 0.000150

 BaP 20 μM 26 Mutants 1 × 10

5 Survivors 0.000256

 BaP 40 μM 84 Mutants 1 × 10

5 Survivors 0.000839

 BaP 60 μM 131 Mutants 1 × 10

5 Survivors 0.001308

 CPcdP 20 μM 34 Mutants 1 × 10

5 Survivors 0.000337

 CPcdP 40 μM 133 Mutants 1 × 10

5 Survivors 0.001330

 ACEP 10 μM 11 Mutants 1 × 10

5 Survivors 0.000110

 ACEP 40 μM 25 Mutants 1 × 10

5 Survivors 0.000248

 ACEP 120 μM 55 Mutants 1 × 10

5 Survivors 0.000551

9620 Chang et al., 2002 TA100 Rat ArS9 Control 0 μg/plate 326 Revertants/

plate 

 SD not 

consistently 

plotted; extract

only point estimdata 

 BaP 5 μg/plate 2,543 Revertants/

plate 

 BghiF 5 μg/plate 1,630 Revertants/

plate 
C-36 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 BcPH 10 μg/plate 1,043 Revertants/

plate 

24030 De Flora et al., 

1984 

Rat AR S9 

 BaP 185 Revertants/

nmol (potency) 

 BaA 12 Revertants/

nmol (potency) 

 Pery 21 Revertants/

nmol (potency) 

 BeP 1.6 Revertants/

nmol (potency) 

18050 Eisenstadt and 

Gold, 1978 

TA100 Rat ArS9 BaP 2 μg 1,705 Revertants/

plate 

 Background 

subtracted from data reported 

 CPcdP 1 μg 134 Revertants/

plate 

18180 Florin et al., 1980 TA100 Rat MC S9 BaP 0.0030 μmol/plate 255 Revertants/

plate 

 Background 

subtracted from data reported 

 TA100 BaA 0.10 μmol/plate 326 Revertants/

plate 

 Only peak 

response report

 TA100 CH 0.0050 μmol/plate 196 Revertants/

plate 

 TA98 BaP 0.0030 μmol/plate 235 Revertants/

plate 

 TA98 Pery 0.025 μmol/plate 91 Revertants/

plate 

 TA98 CO 0.070 μmol/plate 82 Revertants/

plate 

24080 Gibson et al., 1978 TA98 60Co 

gamma radiation, for 7 d (2.5x10

7

rad) 

Control 0 μg/plate 0 Increase in 

revertants 

 Continuous dat

no SD 
C-37 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 BaP 10 μg/plate 1.5 Increase in 

revertants 

 BaP 20 μg/plate 3 Increase in 

revertants 

 BaP 50 μg/plate 10 Increase in 

revertants 

 BaP 100 μg/plate 15 Increase in 

revertants 

 BaP 200 μg/plate 21 Increase in 

revertants 

 BaP 300 μg/plate 35 Increase in 

revertants 

 BaA 150 μg/plate 1.8 Increase in 

revertants 

 BaA 250 μg/plate 6.4 Increase in 

revertants 

 BghiP 400 μg/plate 4.2 Increase in 

revertants 

 CH 500 μg/plate 6.1 Increase in 

revertants 

 CH 1,000 μg/plate 6.7 Increase in 

revertants 

 FE 200 μg/plate 1.1 Increase in 

revertants 

 FE 360 μg/plate 2.2 Increase in 

revertants 

 Pyr 160 μg/plate 28 Increase in 

revertants 

14080 Gold and 

Eisenstadt, 1980 

TA100 50ul rat 

MC S9 

BaP 4 nmol 1,103 Revertants/

plate 

 Background 

subtracted from data reported 

 CPcdP 4 nmol 281 Revertants/

plate 

18650 Hermann, 1981 TA98 Rat Ar S9 BaP 100 Revertants/

nmol (potency) 
C-38 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 BbA 8 Revertants/

nmol (potency) 

 BaA 4 Revertants/

nmol (potency) 

 CH 2 Revertants/

nmol (potency) 

 FA 3 Revertants/

nmol (potency) 

 Tphen 13 Revertants/

nmol (potency) 

 BeP 15 Revertants/

nmol (potency) 

 DBacA 42 Revertants/

nmol (potency) 

 DBahA 8 Revertants/

nmol (potency) 

 BbF 15 Revertants/

nmol (potency) 

 Pery 31 Revertants/

nmol (potency) 

 DBalP 21 Revertants/

nmol (potency) 

 DBaiP 38 Revertants/

nmol (potency) 

 AA 62 Revertants/

nmol (potency) 
C-39 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 CO 60 Revertants/

nmol (potency) 

10670 Johnsen et al., 1997 TA98 PCB 

microsomes 

Control 0 μg/plate 113 Revertants/

plate 

3 8.54 Control respon

added back to eresponse for modeling 

 BaP 10 μg/plate 128 Revertants/

plate 

3 3.66 

 BaP 20 μg/plate 123 Revertants/

plate 

3 13.41 

 BjAC 10 μg/plate 192 Revertants/

plate 

3 10.98 

 BjAC 20 μg/plate 213 Revertants/

plate 

3 9.76 

 BIAC 10 μg/plate 204 Revertants/

plate 

3 13.41 

 BIAC 20 μg/plate 207 Revertants/

plate 

3 43.90 

19000 Kaden et al., 1979 TM677 ArS9 and 

PB S9 

BaP 1 RPF Mutagenic activ

relative to that the 80 μmol Ba

positive control performed 

simultaneously with test compound. 

 AN NA 0.010 RPF  ANL NA 0.070 RPF  Pyr NA 0.070 RPF  BbFE NA 0.080 RPF  CPcdP NA 1.5 RPF  BaA NA 0.14 RPF  CH NA 0.20 RPF  Tphen NA 0.070 RPF  FA NA 1.0 RPF  BeP NA 0.11 RPF 
C-40 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 Pery NA 6 RPF  BghiP NA 0.080 RPF  AA NA 0.080 RPF  DBacA NA 0.77 RPF  DBahA NA 0.080 RPF  DBbeF NA 0.88 RPF 24680 Lafleur et al., 1993 TM677 Rat AR S9 BaP 0 μg/mL 7 Mutants 100,000 Survivors 0.000070 BaP 0.5 μg/mL 8 Mutants 100,000 Survivors 0.000080 BaP 1 μg/mL 10 Mutants 100,000 Survivors 0.000101 BaP 2 μg/mL 18 Mutants 100,000 Survivors 0.000175 BaP 4 μg/mL 22 Mutants 100,000 Survivors 0.000220 BaP 8 μg/mL 33 Mutants 100,000 Survivors 0.000327 BghiF 0 μg/mL 11 Mutants 100,000 Survivors 0.00011  BghiF 1 μg/mL 10 Mutants 100,000 Survivors 0.00010  BghiF 3 μg/mL 14 Mutants 100,000 Survivors 0.00014  BghiF 10 μg/mL 55 Mutants 100,000 Survivors 0.00055  CPcdP 0 μg/mL 12 Mutants 100,000 Survivors 0.000120 CPcdP 0.5 μg/mL 15 Mutants 100,000 Survivors 0.000146 CPcdP 1 μg/mL 13 Mutants 100,000 Survivors 0.000130 CPcdP 2 μg/mL 17 Mutants 100,000 Survivors 0.000172 CPcdP 4 μg/mL 27 Mutants 100,000 Survivors 0.000274
C-41 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 CPcdP 8 μg/mL 60 Mutants 100,000 Survivors 0.000597 CPhiACE

A 

0 μg/mL 8 Mutants 100,000 Survivors 0.000084

 CPhiACE

A 

0.5 μg/mL 10 Mutants 100,000 Survivors 0.000103

 CPhiACE

A 

1 μg/mL 16 Mutants 100,000 Survivors 0.000157

 CPhiACE

A 

2 μg/mL 29 Mutants 100,000 Survivors 0.000286

 CPhiACE

A 

4 μg/mL 67 Mutants 100,000 Survivors 0.000670

 CPhiAPA 0 μg/mL 9 Mutants 100,000 Survivors 0.000090 CPhiAPA 10 μg/mL 12 Mutants 100,000 Survivors 0.000117 CPhiAPA 30 μg/mL 21 Mutants 100,000 Survivors 0.000210 CPhiAPA 100 μg/mL 26 Mutants 100,000 Survivors 0.000263 ACEA 0 μg/mL 9 Mutants 100,000 Survivors 0.000092 ACEA 10 μg/mL 21 Mutants 100,000 Survivors 0.000214 ACEA 35 μg/mL 69 Mutants 100,000 Survivors 0.000686 APA 0 μg/mL 16 Mutants 100,000 Survivors 0.000160 APA 10 μg/mL 37 Mutants 100,000 Survivors 0.000375 APA 30 μg/mL 42 Mutants 100,000 Survivors 0.000416 APA 100 μg/mL 22 Mutants 100,000 Survivors 0.00022019320 LaVoie et al., 1979 TA98 Rat Ar S9 BaP 10 μg 450 Revertants/

plate 

 Background 

subtracted from data reported 
C-42 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 BaP 20 μg 480 Revertants/

plate 

 BeP 10 μg 20 Revertants/

plate 

 BeP 20 μg 20 Revertants/

plate 

 Pery 20 μg 70 Revertants/

plate 

23650 McCann et al., 

1975 

Multiple strains 

Rat Ar S9 BaP NA 121 Revertants/ 

nmol (potency) 

 Paper states tha

comparison of 

potency estima

should be done with caution (nlinear doseresponse) see tafootnotes 

 DBaiP NA 20 Revertants/

nmol (potency) 

 BeP NA 0.6 Revertants/

nmol (potency) 

 DBacA NA 175 Revertants/

nmol (potency) 

 DBahA NA 11 Revertants/

nmol (potency) 

 CH NA 38 Revertants/

nmol (potency) 

 BaA NA 11 Revertants/

nmol (potency) 

20220 Pahlman and 

Pelkonen, 1987 

TA100 Rat MC S9 BaP NA 272 Revertants/

nmol (potency) 
C-43 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 BaA NA 10.4 Revertants/

nmol (potency) 

 CH NA 9.7 Revertants/

nmol (potency) 

 Tphen NA 4 Revertants/

nmol (potency) 

 DBacA NA 35 Revertants/

nmol (potency) 

 DBahA NA 4.4 Revertants/

nmol (potency) 

20450 Phillipson and 

Ioannides, 1989 

TA100 Hamster S9 BaP 0 μg/plate 0.000 Revertants/

plate 

 BaP 5 μg/plate 68.833 Revertants/

plate 

 BaP 10 μg/plate 118.948 Revertants/

plate 

 BaP 15 μg/plate 99.744 Revertants/

plate 

 BaP 20 μg/plate 96.101 Revertants/

plate 

 BaA 0 μg/plate 0.000 Revertants/

plate 

 BaA 20 μg/plate 109.877 Revertants/

plate 

 BaA 40 μg/plate 115.248 Revertants/

plate 

 BaA 60 μg/plate 114.430 Revertants/

plate 

 BaA 100 μg/plate 98.846 Revertants/

plate 

 DBaiP 0 μg/plate 0.000 Revertants/

plate 
C-44 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 DBaiP 20 μg/plate 64.638 Revertants/

plate 

 DBaiP 40 μg/plate 75.747 Revertants/

plate 

 DBaiP 60 μg/plate 80.394 Revertants/

plate 

 DBaiP 100 μg/plate 63.880 Revertants/

plate 

 DBahA 0 μg/plate 0.000 Revertants/

plate 

 DBahA 10 μg/plate 50.899 Revertants/

plate 

 DBahA 20 μg/plate 56.886 Revertants/

plate 

 DBahA 30 μg/plate 52.419 Revertants/

plate 

 DBahA 50 μg/plate 34.980 Revertants/

plate 

21000 Sakai et al., 1985 TA97 Rat Ar S9 Control 0 μg 177 Revertants/

plate 

 BaP 1 μg 1,208 Revertants/

plate 

 BaP 5 μg 1,432 Revertants/

plate 

 BaP 10 μg 1,742 Revertants/

plate 

 Control 0 μg 189 Revertants/

plate 

 FE 5 μg 254 Revertants/

plate 

 FE 10 μg 240 Revertants/

plate 

 FE 50 μg 240 Revertants/

plate 

 FE 250 μg 232 Revertants/

plate 
C-45 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 Control 0 μg 189 Revertants/

plate 

 AC 5 μg 360 Revertants/

plate 

 AC 10 μg 509 Revertants/

plate 

 AC 50 μg 293 Revertants/

plate 

 AC 250 μg 279 Revertants/

plate 

 Control 0 μg 189 Revertants/

plate 

 PH 5 μg 454 Revertants/

plate 

 PH 10 μg 534 Revertants/

plate 

 PH 50 μg 321 Revertants/

plate 

 PH 250 μg T Revertants/

plate 

 Control 0 μg 177 Revertants/

plate 

 FA 5 μg 652 Revertants/

plate 

 FA 10 μg 1,012 Revertants/

plate 

 FA 50 μg 1,042 Revertants/

plate 

 FA 250 μg 518 Revertants/

plate 

 Control 0 μg 177 Revertants/

plate 

 CH 5 μg 640 Revertants/

plate 

 CH 10 μg 815 Revertants/

plate 
C-46 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 CH 20 μg 888 Revertants/

plate 

 CH 50 μg 723 Revertants/

plate 

 Control 0 μg 177 Revertants/

plate 

 Pyr 2 μg 929 Revertants/

plate 

 Pyr 4 μg 1,582 Revertants/

plate 

 Pyr 6 μg 2,057 Revertants/

plate 

 Pyr 10 μg 2,577 Revertants/

plate 

 Pyr 20 μg 2,832 Revertants/

plate 

 Pyr 50 μg 2,296 Revertants/

plate 

 Control 0 μg 177 Revertants/

plate 

 BeP 5 μg 944 Revertants/

plate 

 BeP 10 μg 1,100 Revertants/

plate 

 BeP 50 μg 606 Revertants/

plate 

 BeP 250 μg 640 Revertants/

plate 

 Control 0 μg 177 Revertants/

plate 

 Pery 1 μg 1,516 Revertants/

plate 

 Pery 2 μg 2,236 Revertants/

plate 

 Pery 4 μg 2,784 Revertants/

plate 
C-47 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 Pery 10 μg 2,550 Revertants/

plate 

 Pery 50 μg 1,808 Revertants/

plate 

 Control 0 μg 177 Revertants/

plate 

 BghiP 10 μg 896 Revertants/

plate 

 BghiP 20 μg 991 Revertants/

plate 

 BghiP 50 μg 896 Revertants/

plate 

 BghiP 250 μg 612 Revertants/

plate 

 Control 0 μg 177 Revertants/

plate 

 CO 5 μg 362 Revertants/

plate 

 CO 10 μg 400 Revertants/

plate 

 CO 50 μg 405 Revertants/

plate 

 CO 100 μg 490 Revertants/

plate 

 CO 200 μg 479 Revertants/

plate 

11860 Sangaiah et al., 

1983 

TA98 Rat Ar S9 Control 0 μg/plate 35.43 Revertants/

plate 

 BaP 2 μg/plate 177.37 Revertants/

plate 

 BaP 3 μg/plate 266.02 Revertants/

plate 

 BaP 6 μg/plate 419.68 Revertants/

plate 

 BaP 10 μg/plate 312.76 Revertants/

plate 
C-48 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 BaP 30 μg/plate 358.41 Revertants/

plate 

 BaP 50 μg/plate 350.92 Revertants/

plate 

 BaP 100 μg/plate 323.12 Revertants/

plate 

 Control 0 μg/plate 53.15 Revertants/

plate 

 BjAC 2 μg/plate 124.15 Revertants/

plate 

 BjAC 3 μg/plate 331.10 Revertants/

plate 

 BjAC 6 μg/plate 674.11 Revertants/

plate 

 BjAC 10 μg/plate 993.21 Revertants/

plate 

 BjAC 30 μg/plate 1,027.06 Revertants/

plate 

 BjAC 50 μg/plate 883.45 Revertants/

plate 

 BjAC 100 μg/plate 1,021.36 Revertants/

plate 

21360 Simmon, 1979a TA100 Rat Ar S9 BaP 5 μg 1,141 Revertants/

plate 

 Background 

subtracted from data reported 

 BaA 50 μg 280 Revertants/

plate 

 BeP 50 μg 57 Revertants/

plate 

21640 Teranishi et al., 

1975 

TA1538 Rat PB S9 Control 0 μg/plate 38 Revertant 

colonies/

plate 

 BaP 50 μg/plate 77 Revertant 

colonies/

plate 
C-49 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 DBaiP 50 μg/plate 102 Revertant 

colonies/

plate 

 TA1538 Rat PB and 

DBahA S9 

Control 0 μg/plate 25 Revertant 

colonies/

plate 

 BaP 50 μg/plate 279 Revertant 

colonies/

plate 

 DBaeP 50 μg/plate 88 Revertant 

colonies/

plate 

16180 Utesch et al., 1987 TA100 With 

homogenized hepatocytes 

from Ar treated rats 

Control 0 μg/plate 159 Revertants/

plates 

 BaP 6.3 μg/plate 998 Revertants/

plate 

 BaP 12.5 μg/plate 1,079 Revertants/

plate 

 BaP 25 μg/plate 1,178 Revertants/

plate 

 BaP 50 μg/plate 1,141 Revertants/

plate 

 BaP 100 μg/plate 1,114 Revertants/

plate 

 Control 0 μg/plate 199 Revertants/

plate 

 BaA 6.3 μg/plate 861 Revertants/

plate 

 BaA 12.5 μg/plate 2,583 Revertants/

plate 

 BaA 25 μg/plate 3,546 Revertants/

plate 

 BaA 50 μg/plate 3,786 Revertants/

plate 
C-50 DRAFT – DO NOT CITE OR QUO

Table C-7. In vitro bacterial mutagenicity: dose response data 

Record 

number Reference Cell type PAH Dose Dose units Response

Response 

units n Units

% response Std dev Std error Comments 

 BaA 100 μg/plate 3,406 Revertants/

plate 

16440 Wood et al., 1980 TA98 Purified 

microsomal P450 

Control 0 nmol 0 Revertants/

plate 

 Background 

subtracted from data reported 

 BaP 3.75 nmol 45 Revertants/

plate 

 BaP 7.5 nmol 63 Revertants/

plate 

 BaP 15 nmol 99 Revertants/

plate 

 BaP 30 nmol 103 Revertants/

plate 

 Control 0 nmol 0 Revertants/

plate 

 CPcdP 3.75 nmol 303 Revertants/

plate 

 CPcdP 7.5 nmol 491 Revertants/

plate 

 CPcdP 15 nmol 685 Revertants/

plate 

 CPcdP 30 nmol 776 Revertants/

plate 

1 
C-51 DRAFT – DO NOT CITE OR QUOTE 

Table C-8. In vitro mammalian mutagenicity: data use 

Record 

number Reference 

Data 

source Data points 

Basis for RPF 

approach Comments 

16920 Amacher and 

Paillet, 1982 

Figure 1 Use lines for BaP (open 

circles) and BaA (closed 

triangles; dose is μg/mL and 

response is mutation 

frequency (MF)/106 survivors

Model; 

quantal data 

Thymidine kinase assay (resistance to trifluorothymidine) in 

mouse lymphoma cells 

(L5178Y) with Syrian golden 

hamster S9 mix or cocultivated 

hamster hepatocytes 

16940 Amacher and 

Turner, 1980

Figure 3 Use bars for SM2 S9 

activation for BaP and BaA; 

dose is 1.25 × 10-5 M for BaP 

and 3.22 × 10-5 M for BaP; 

response is IMF/104

 survivors

Point estimate Thymidine kinase assay 

(resistance to trifluorothymidine) in mouse lymphoma cells 

(L5178Y) with mouse S9 mix 

16910 Amacher et 

al., 1980 

Table 3 Use DR data for BaA and 

BaP; dose as concentration 

(M), response as mutants per 

104 survivors 

Model; 

quantal data 

Thymidine kinase assay 

(resistance to trifluorothymidine) in mouse lymphoma cells 

(L5178Y) with mouse S9 mix 

17140 Barfknecht et 

al., 1982 

Figure 2 

(BaP, FA); 

Figure 4 

(BaA, CH, 

Tphen); 

Figure 6 

(CPcdP) 

Dose is μM and mutant 

fraction ×106 

Model; 

quantal data 

Thymidine kinase assay 

(resistance to trifluorothymidine) in human lymphoblast 

cells with rat Ar S9 mix 

14250 Hass et al., 

1982 

Table 1 DR data for DBaiP, DBahP, 

and BaP; dose is μg/mL; use 

response data for TG mutants 

only (mutants/106 cells); 

control value is 4 ± 

1 mutants/106

 cells 

Model; 

quantal data 

Hypoxanthine-guanine phosphoribosyl transferase assay 

(resistance to 6-thioguanine) in 

V79 Chinese hamster cells 

with rat MC S9 

18740 Huberman 

and Sachs, 

1976 

Table 2 Use data for BaP, DBacA, 

DBahA; 8-azaguanine 

resistance only; use 1μg/mL 

dose for all (*), response as 

mutants per 105 survivors 

Point estimate Hypoxanthine-guanine phosphoribosyl transferase assay 

(resistance to 8-azaguanine) in 

V79 Chinese hamster cells 

with hamster embryo cells 

18990 Jotz and 

Mitchell, 

1981 

Table 2 Use data for BaP and Pyr with 

metabolic activation; subtract 

negative control, dose as 

μg/mL, response as MF × 10-6

Point estimate Thymidine kinase assay 

(resistance to trifluorothymidine) in mouse lymphoma cells 

(L5178Y) with rat Ar S9 

24720 Kligerman et 

al., 1986 

Figure 1 Use DR dat for BaP and 

BlAC; dose as μg/mL, 

response as mutant 

frequency/106

 survivors; 

average data from two 

experiments 

Model; 

quantal data 

Thymidine kinase assay 

(resistance to trifluorothymidine) in mouse lymphoma cells 

(L5178Y) with rat Ar S9 

19180 Krahn and 

Heidelberger, 

1977 

Table II Use data for BaP, DBahA, 

DBacA, and BaA; cell 

survival @40% control 

(column 3), controls are 100% 

survival group (column 1); 

use 3-MC S9 data only; dose 

as nmol/mL, response as 

6-TG/105 cells 

Point estimate Hypoxanthine-guanine 

phosphoribosyl transferase 

assay (resistance to 6-thioguanine) in V79 Chinese 

hamster cells with hamster 

embryo cells 
C-52 DRAFT – DO NOT CITE OR QUOTE 

Table C-8. In vitro mammalian mutagenicity: data use 

Record 

number Reference 

Data 

source Data points 

Basis for RPF 

approach Comments 

24680 Lafleur et al., 

1993 

Figures 5 

and 6 

Use DR curves for BaP, 

CPcdP (CPP), CPhiACEA 

(CPAA), ACEA (AA); dose 

as μg/mL, response as mutant 

fraction (ppm) 

Model as 

quantal data 

(mutant 

fraction 

reported) 

Thymidine kinase assay (resistance to trifluorothymidine) in 

MCL-3 cells (human Blymphoblastoid cells) 

7550 Li and Lin, 

1996 

Text Mutant frequency of controls 

2 × 10-5; 10 ng/mL BaP = 5 × 

10-5; BaA = 5.6 × 10-5

Point estimate Hypoxanthine-guanine phosphoribosyl transferase assay 

(resistance to 6-thioguanine) in 

HS1 HeLa cells (human 

epithelial cells) 

11450 Nesnow et 

al., 1984 

Chart 9 Use data for BaP, BlAC, 

BeAC, and BjAC; dose as 

μg/mL, response as 

6TG-resistant mutants/ 

106 survivors 

Model; 

quantal data 

Hypoxanthine-guanine phosphoribosyl transferase assay 

(resistance to 6-thioguanine) in 

V79 Chinese hamster cells 

with rat AR S9 

15630 Raveh and 

Huberman, 

1983 

Table 1 Use data for CPcdP and BaP, 

with PMA only; dose in 

μg/mL, response in 

mutants/105

 cells 

Model; 

quantal data 

Hypoxanthine-guanine 

phosphoribosyl transferase 

assay (resistance to 6-thioguanine) in V79 Chinese 

hamster cells with hamster 

embryo cells 

15640 Raveh et al., 

1982 

Figure 4 Use DR data for CPcdP and 

BaP (ouabain resistance only); 

dose in μg/mL, response in 

mutants/106 cells 

Model; 

quantal data 

Hypoxanthine-guanine phosphoribosyl transferase assay 

(resistance to ouabain) in V79 

Chinese hamster cells with 

hamster embryo cells 

21410 Slaga et al., 

1978 

Table 3 Use DR data for BaA and 

BaP; dose as μM, response as 

ouabain resistant 

mutants/104 survivors 

Model; 

quantal data 

Hypoxanthine-guanine phosphoribosyl transferase assay 

(resistance to ouabain) in V79 

Chinese hamster cells with 

hamster embryo cells 

16190 Vaca et al., 

1992 

Figure 5 DR data for FA and BaP; dose 

as μM, response as 6-Tg 

resistant cells/100,000 

Model; 

quantal data 

Hypoxanthine-guanine 

phosphoribosyl transferase 

assay (resistance to 6-thioguanine) in UV-sensitive CHO 

cells with rat Ar S9 

21900 Wangenheim 

and 

Bolcsfoldi, 

1988 

Table 1 Use +S9 DR data for Pyr, 

BaP, and FE; dose as mol/L, 

response as mutation 

frequency 

Model; 

quantal data 

Thymidine kinase assay 

(resistance to trifluorothymidine) in mouse lymphoma cells (L5178Y) with rat 

Ar S9 

24670 Durant et al., 

1999 

Table 1 Use DR data for BaPery, 

BbPery, DBaeF, DBafF, 

DBahP, DBaiP, DBelP, 

N23aP, N23eP; positive 

control is reported as 

1,000 ng/mL BaP (reported 

separately for each PAH) 

Model; 

quantal data 

Thymidine kinase assay 

(resistance to trifluorothymidine) in human h1Alv2 

cells 
C-53 DRAFT – DO NOT CITE OR QUO

Table C-9. In vitro mammalian mutagenicity: dose response data 

Record 

number Reference PAH Dose Dose units Mutants In number Units % response Comments 

16920 Amacher and 

Paillet, 1982 

Control 0 μg/mL 39 1 × 10

6 Survivors 0.000039 

 BaP 2.5 μg/mL 119 1 × 10

6 Survivors 0.00012 

 BaP 5 μg/mL 170 1 × 10

6 Survivors 0.00017 

 BaP 7.5 μg/mL 196 1 × 10

6 Survivors 0.00020 

 BaP 10 μg/mL 267 1 × 10

6 Survivors 0.00027 

 Control 0 μg/mL 20 1 × 10

6 Survivors 0.000020 

 BaA 2.5 μg/mL 65 1 × 10

6 Survivors 0.000065 

 BaA 5 μg/mL 62 1 × 10

6 Survivors 0.000062 

 BaA 10 μg/mL 88 1 × 10

6 Survivors 0.000088 

 BaA 15 μg/mL 89 1 × 10

6 Survivors 0.000089 

16940 Amacher and 

Turner, 1980 

Control 0 M 0.4 1 × 10

4 Survivors 0.000040 Control w/o S9 treatm

 BaP 1.25 × 10-5 M 2.85 1 × 10

4 Survivors 0.000285 

 BaA 3.22 × 10-5 M 3.12 1 × 10

4 Survivors 0.000312 

16910 Amacher et al., 

1980 

Control 0 M 0.680 1 × 10

4 Survivors 0.000068 

 BaP 5.30 × 10-6 M 1.360 1 × 10

4 Survivors 0.000136 

 BaP 7.00 × 10-6 M 1.790 1 × 10

4 Survivors 0.000179 

 BaP 9.40 × 10-6 M 1.470 1 × 10

4 Survivors 0.000147 

 BaP 1.25 × 10-5 M 1.870 1 × 10

4 Survivors 0.000187 

 BaP 1.67 × 10-5 M 2.600 1 × 10

4 Survivors 0.000260 

 BaP 2.23 × 10-5 M 2.490 1 × 10

4 Survivors 0.000249 

 BaP 2.97 × 10-5 M 2.650 1 × 10

4 Survivors 0.000265 

 BaP 3.96 × 10-5 M 3.970 1 × 10

4 Survivors 0.000397 

 Control 0 M 0.770 1 × 10

4 Survivors 0.000077 

 BaA 1.36 × 10-5 M 0.810 1 × 10

4 Survivors 0.000081 

 BaA 1.81 × 10-5 M 0.840 1 × 10

4 Survivors 0.000084 

 BaA 2.42 × 10-5 M 1.000 1 × 10

4 Survivors 0.000100 

 BaA 3.22 × 10-5 M 1.230 1 × 10

4 Survivors 0.000123 

 BaA 4.30 × 10-5 M 1.470 1 × 10

4 Survivors 0.000147 

 BaA 5.47 × 10-5 M NS 1 × 10

4 Survivors NS = no survivors 

 BaA 7.65 × 10-5 M NS 1 × 10

4 Survivors 
C-54 DRAFT – DO NOT CITE OR QUO

Table C-9. In vitro mammalian mutagenicity: dose response data 

Record 

number Reference PAH Dose Dose units Mutants In number Units % response Comments 

 BaA 1.02 × 10-4 M NS 1 × 10

4 Survivors 

17140 Barfknecht et al., 

1982 

Control 0 μM 0 1 × 10

6 Survivors 0.000000 

 BaP 10 μM 51 1 × 10

6 Survivors 0.000051 

 BaP 20 μM 120 1 × 10

6 Survivors 0.000120 

 BaP 30 μM 155 1 × 10

6 Survivors 0.000155 

 Control 0 μM 0 1 × 10

6 Survivors 0.000000 

 FA 10 μM 27 1 × 10

6 Survivors 0.000027 

 FA 20 μM 50 1 × 10

6 Survivors 0.000050 

 FA 40 μM 62 1 × 10

6 Survivors 0.000062 

 Control 0 μM 0 1 × 10

6 Survivors 0.000000 

 BaA 20 μM 12 1 × 10

6 Survivors 0.000012 

 BaA 50 μM 29 1 × 10

6 Survivors 0.000029 

 BaA 100 μM 34 1 × 10

6 Survivors 0.000034 

 BaA 150 μM 64 1 × 10

6 Survivors 0.000064 

 Control 0 μM 0 1 × 10

6 Survivors 0.000000 

 CH 20 μM 17 1 × 10

6 Survivors 0.000017 

 CH 50 μM 26 1 × 10

6 Survivors 0.000026 

 CH 100 μM 30 1 × 10

6 Survivors 0.000030 

 Control 0 μM 0 1 × 10

6 Survivors 0.000000 

 Tphen 50 μM 10 1 × 10

6 Survivors 0.000010 

 Tphen 100 μM 20 1 × 10

6 Survivors 0.000020 

 Tphen 200 μM 35 1 × 10

6 Survivors 0.000035 

 Control 0 μM 3 1 × 10

6 Survivors 0.000003 

 CPcdP 23 μM 11 1 × 10

6 Survivors 0.000011 

 CPcdP 47 μM 24 1 × 10

6 Survivors 0.000024 

 CPcdP 88 μM 27 1 × 10

6 Survivors 0.000027 

24670 Durant et al., 1999 BaP 1,000 ng/mL 170 1 × 10

6 Survivors 0.00017 

 BaP 1,000 ng/mL 170 1 × 10

6 Survivors 0.00017 

 BaP 1,000 ng/mL 200 1 × 10

6 Survivors 0.00020 

 BaP 1,000 ng/mL 200 1 × 10

6 Survivors 0.00020 

 BaP 1,000 ng/mL 160 1 × 10

6 Survivors 0.00016 
C-55 DRAFT – DO NOT CITE OR QUO

Table C-9. In vitro mammalian mutagenicity: dose response data 

Record 

number Reference PAH Dose Dose units Mutants In number Units % response Comments 

 BaP 1,000 ng/mL 170 1 × 10

6 Survivors 0.00017 

 BaP 1,000 ng/mL 190 1 × 10

6 Survivors 0.00019 

 BaP 1,000 ng/mL 200 1 × 10

6 Survivors 0.00020 

 BaP 1,000 ng/mL 210 1 × 10

6 Survivors 0.00021 

 Averaged 

BaP 

1,000 ng/mL 186 1 × 10

6 Survivors 0.00019 

 Averaged 

controls 

0 ng/mL 20 1 × 10

6 Survivors 0.00002 

 Control 0 ng/mL 18 1 × 10

6 Survivors 0.000018 

 BaPery 0.1 ng/mL 21 1 × 10

6 Survivors 0.000021 

 BaPery 0.3 ng/mL 23 1 × 10

6 Survivors 0.000023 

 BaPery 1 ng/mL 28 1 × 10

6 Survivors 0.000028 

 BaPery 3 ng/mL 50 1 × 10

6 Survivors 0.000050 

 BaPery 10 ng/mL 82 1 × 10

6 Survivors 0.000082 

 BaPery 100 ng/mL 200 1 × 10

6 Survivors 0.00020 

 Control 0 ng/mL 18 1 × 10

6 Survivors 0.000018 

 BbPery 1 ng/mL 19 1 × 10

6 Survivors 0.000019 

 BbPery 3 ng/mL 22 1 × 10

6 Survivors 0.000022 

 BbPery 10 ng/mL 32 1 × 10

6 Survivors 0.000032 

 BbPery 100 ng/mL 54 1 × 10

6 Survivors 0.000054 

 Control 0 ng/mL 21 1 × 10

6 Survivors 0.000021 

 DBaeF 1 ng/mL 29 1 × 10

6 Survivors 0.000029 

 DBaeF 10 ng/mL 72 1 × 10

6 Survivors 0.000072 

 DBaeF 100 ng/mL 190 1 × 10

6 Survivors 0.00019 

 DBaeF 1,000 ng/mL np 1 × 10

6 Survivors Not plated due to 

excessive toxicity 

 Control 0 ng/mL 21 1 × 10

6 Survivors 0.000021 

 DBafF 1 ng/mL 21 1 × 10

6 Survivors 0.000021 

 DBafF 10 ng/mL 37 1 × 10

6 Survivors 0.000037 

 DBafF 100 ng/mL 81 1 × 10

6 Survivors 0.000081 

 DBafF 1,000 ng/mL 190 1 × 10

6 Survivors 0.00019 

 Control 0 ng/mL 19 1 × 10

6 Survivors 0.000019 

 DBahP 0.1 ng/mL 24 1 × 10

6 Survivors 0.000024 
C-56 DRAFT – DO NOT CITE OR QUO

Table C-9. In vitro mammalian mutagenicity: dose response data 

Record 

number Reference PAH Dose Dose units Mutants In number Units % response Comments 

 DBahP 1 ng/mL 24 1 × 10

6 Survivors 0.000024 

 DBahP 10 ng/mL 46 1 × 10

6 Survivors 0.000046 

 DBahP 100 ng/mL 80 1 × 10

6 Survivors 0.000080 

 Control 0 ng/mL 20 1 × 10

6 Survivors 0.000020 

 DBaiP 0.3 ng/mL 20 1 × 10

6 Survivors 0.000020 

 DBaiP 1 ng/mL 35 1 × 10

6 Survivors 0.000035 

 DBaiP 10 ng/mL 88 1 × 10

6 Survivors 0.000088 

 DBaiP 100 ng/mL 150 1 × 10

6 Survivors 0.00015 

 Control 0 ng/mL 21 1 × 10

6 Survivors 0.000021 

 DBelP 10 ng/mL 28 1 × 10

6 Survivors 0.000028 

 DBelP 100 ng/mL 34 1 × 10

6 Survivors 0.000034 

 DBelP 1,000 ng/mL 55 1 × 10

6 Survivors 0.000055 

 Control 0 ng/mL 21 1 × 10

6 Survivors 0.000021 

 N23aP 0.1 ng/mL 23 1 × 10

6 Survivors 0.000023 

 N23aP 1 ng/mL 44 1 × 10

6 Survivors 0.000044 

 N23aP 10 ng/mL 84 1 × 10

6 Survivors 0.000084 

 N23aP 100 ng/mL 94 1 × 10

6 Survivors 0.000094 

 N23aP 1,000 ng/mL 73 1 × 10

6 Survivors 0.000073 

 Control 0 ng/mL 19 1 × 10

6 Survivors 0.000019 

 N23eP 1 ng/mL 20 1 × 10

6 Survivors 0.000020 

 N23eP 10 ng/mL 41 1 × 10

6 Survivors 0.000041 

 N23eP 100 ng/mL 74 1 × 10

6 Survivors 0.000074 

 N23eP 1,000 ng/mL 98 1 × 10

6 Survivors 0.00010 

14250 Hass et al., 1982 Control 0 μg/mL 4 1 × 10

6 CFC 0.0000040 

 BaP 0.30 μg/mL 267 1 × 10

6 CFC 0.00027 

 BaP 1.00 μg/mL 293 1 × 10

6 CFC 0.00029 

 DBaiP 0.03 μg/mL 124 1 × 10

6 CFC 0.00012 

 DBaiP 0.10 μg/mL 289 1 × 10

6 CFC 0.00029 

 DBaiP 0.30 μg/mL 1211 1 × 10

6 CFC 0.00121 

 DBahP 0.03 μg/mL 110 1 × 10

6 CFC 0.00011 

 DBahP 0.10 μg/mL 264 1 × 10

6 CFC 0.00026 

 DBahP 0.30 μg/mL 668 1 × 10

6 CFC 0.00067 
C-57 DRAFT – DO NOT CITE OR QUO

Table C-9. In vitro mammalian mutagenicity: dose response data 

Record 

number Reference PAH Dose Dose units Mutants In number Units % response Comments 

18740 Huberman and 

Sachs, 1976 

Control 0 μg/mL 6 1 × 10

5 Survivors 0.000060 

 BaP 1 μg/mL 425 1 × 10

5 Survivors 0.00425 

 DBacA 1 μg/mL 22 1 × 10

5 Survivors 0.00022 

 DBahA 1 μg/mL 17 1 × 10

5 Survivors 0.00017 

18990 Jotz and Mitchell, 

1981 

Control 0 μg/mL 80 1 × 10

6 Survivors 0.000080 

 BaP 4.5 μg/mL 224 1 × 10

6 Survivors 0.00022 With metabolic 

activation 

 Control 0 μg/mL 116 1 × 10

6 Survivors 0.00012 

 Pyr 10.6 μg/mL 150 1 × 10

6 Survivors 0.00015 With metabolic 

activation 

24720 Kligerman et al., 

1986 

Control 0 nmol/mL 92 1 × 10

6 Survivors 0.00009 Avg. of 2 experiments 

 BaP 2.0 nmol/mL 258 1 × 10

6 Survivors 0.00026 

 BaP 3.0 nmol/mL 417 1 × 10

6 Survivors 0.00042 

 BaP 4.0 nmol/mL 557 1 × 10

6 Survivors 0.00056 

 Control 0 nmol/mL 90 1 × 10

6 Survivors 0.00009 

 BlAC 0.5 nmol/mL 93 1 × 10

6 Survivors 0.00009 

 BlAC 2.5 nmol/mL 197 1 × 10

6 Survivors 0.00020 

 BlAC 5.0 nmol/mL 374 1 × 10

6 Survivors 0.00037 

19180 Krahn and 

Heidelberger, 1977 

Control 0 nmol/mL 1.7 1 × 10

5 Survivors 0.000017 

 BaP 15.9 nmol/mL 14 1 × 10

5 Survivors 0.000136 3-MC S9; 40% surviv

 Control 0 nmol/mL 1.5 1 × 10

5 Survivors 0.000015 

 BaA 46.5 nmol/mL 6.5 1 × 10

5 Survivors 0.000065 3-MC S9; 40% surviv

24680 Lafleur et al., 1993 Control 0 μg/mL 1.2 1 × 10

6 Survivors 0.0000012 

 BaP 0.02 μg/mL 4.8 1 × 10

6 Survivors 0.0000048 

 BaP 0.06 μg/mL 24 1 × 10

6 Survivors 0.000024 

 BaP 0.2 μg/mL 25 1 × 10

6 Survivors 0.000025 

 BaP 1 μg/mL 39 1 × 10

6 Survivors 0.000039 

 BaP 5 μg/mL 56 1 × 10

6 Survivors 0.000056 

 Control 0 μg/mL 1.8 1 × 10

6 Survivors 0.0000018 

 ACEA 1 μg/mL 6.0 1 × 10

6 Survivors 0.0000060 
C-58 DRAFT – DO NOT CITE OR QUO

Table C-9. In vitro mammalian mutagenicity: dose response data 

Record 

number Reference PAH Dose Dose units Mutants In number Units % response Comments 

 ACEA 3 μg/mL 15 1 × 10

6 Survivors 0.000015 

 ACEA 8 μg/mL 21 1 × 10

6 Survivors 0.000021 

 Control 0 μg/mL 2.5 1 × 10

6 Survivors 0.0000025 

 CPcdP 0.03 μg/mL 4.2 1 × 10

6 Survivors 0.0000042 

 CPcdP 0.06 μg/mL 4.9 1 × 10

6 Survivors 0.0000049 

 CPcdP 0.2 μg/mL 5.9 1 × 10

6 Survivors 0.0000059 

 CPcdP 0.6 μg/mL 10 1 × 10

6 Survivors 0.000010 

 CPcdP 2 μg/mL 17 1 × 10

6 Survivors 0.000017 

 Control 0 μg/mL 2.8 1 × 10

6 Survivors 0.0000028 

 CPhiACEA 0.1 μg/mL 12 1 × 10

6 Survivors 0.000012 

 CPhiACEA 0.3 μg/mL 25 1 × 10

6 Survivors 0.000025 

 CPhiACEA 0.8 μg/mL 31 1 × 10

6 Survivors 0.000031 

7550 Li and Lin, 1996 Control 0 ng/mL 2 1 × 10

5 Survivors 0.000020 

 BaP 10 ng/mL 5 1 × 10

5 Survivors 0.000050 

 BaA 10 ng/mL 5.6 1 × 10

5 Survivors 0.000056 

11450 Nesnow et al., 1984 Control 0 μg/mL 16 1 × 10

6 Survivors 0.000016 

 BaP 0.5 μg/mL 10 1 × 10

6 Survivors 0.000010 

 BaP 1.0 μg/mL 46 1 × 10

6 Survivors 0.000046 

 BaP 2.5 μg/mL 72 1 × 10

6 Survivors 0.000072 

 BaP 5.0 μg/mL 206 1 × 10

6 Survivors 0.000206 

 BaP 10.0 μg/mL 215 1 × 10

6 Survivors 0.000215 

 BaP 20.0 μg/mL 293 1 × 10

6 Survivors 0.000293 

 BeAC 1.0 μg/mL 17 1 × 10

6 Survivors 0.000017 

 BeAC 2.5 μg/mL 53 1 × 10

6 Survivors 0.000053 

 BeAC 5.0 μg/mL 435 1 × 10

6 Survivors 0.000435 

 BeAC 10.0 μg/mL 235 1 × 10

6 Survivors 0.000235 

 BeAC 20.0 μg/mL 349 1 × 10

6 Survivors 0.000349 

 BjAC 1.0 μg/mL 24 1 × 10

6 Survivors 0.000024 

 BjAC 2.5 μg/mL 94 1 × 10

6 Survivors 0.000094 

 BjAC 5.0 μg/mL 268 1 × 10

6 Survivors 0.000268 

 BjAC 10.0 μg/mL 225 1 × 10

6 Survivors 0.000225 

 BjAC 20.0 μg/mL 215 1 × 10

6 Survivors 0.000215 
C-59 DRAFT – DO NOT CITE OR QUO

Table C-9. In vitro mammalian mutagenicity: dose response data 

Record 

number Reference PAH Dose Dose units Mutants In number Units % response Comments 

 BlAC 1.0 μg/mL 31 1 × 10

6 Survivors 0.000031 

 BlAC 2.5 μg/mL 454 1 × 10

6 Survivors 0.000454 

 BlAC 5.0 μg/mL 320 1 × 10

6 Survivors 0.000320 

 BlAC 10.0 μg/mL 704 1 × 10

6 Survivors 0.000704 

 BlAC 20.0 μg/mL 769 1 × 10

6 Survivors 0.000769 

15630 Raveh and 

Huberman, 1983 

Control 0 μg/mL 3 1 × 10

5 Survivors 0.000030 

 BaP 0.3 μg/mL 25 1 × 10

5 Survivors 0.00025 

 BaP 1 μg/mL 103 1 × 10

5 Survivors 0.0010 

 CPcdP 0.3 μg/mL 9 1 × 10

5 Survivors 0.000090 

 CPcdP 1 μg/mL 20 1 × 10

5 Survivors 0.00020 

15640 Raveh et al., 1982 BaP 0 μg/mL 7 1 × 10

6 CFC 0.0000070 

 BaP 0.3 μg/mL 20 1 × 10

6 CFC 0.000020 

 BaP 1 μg/mL 74 1 × 10

6 CFC 0.000074 

 BaP 3 μg/mL 74 1 × 10

6 CFC 0.000074 

 CPcdP 0 μg/mL 1 1 × 10

6 CFC 0.0000010 

 CPcdP 0.3 μg/mL 5 1 × 10

6 CFC 0.0000047 

 CPcdP 1 μg/mL 10 1 × 10

6 CFC 0.000010 

 CPcdP 3 μg/mL 28 1 × 10

6 CFC 0.000028 

21410 Slaga et al., 1978 Control 0 μM 0.7 1 × 10

4 Survivors 0.000070 

 BaA 4.4 μM 0.9 1 × 10

4 Survivors 0.000090 

 BaA 44.0 μM 2.1 1 × 10

4 Survivors 0.00021 

 BaP 0.4 μM 11.0 1 × 10

4 Survivors 0.0011 

 BaP 1.3 μM 25.0 1 × 10

4 Survivors 0.0025 

 BaP 4.0 μM 99.0 1 × 10

4 Survivors 0.0099 

16190 Vaca et al., 1992 BaP 0 μM 3 1 × 10

5 Survivors 0.000032 

 BaP 2 μM 10 1 × 10

5 Survivors 0.000102 

 BaP 4 μM 23 1 × 10

5 Survivors 0.000229 

 BaP 10 μM 31 1 × 10

5 Survivors 0.000306 

 FA 0 μM 10 1 × 10

5 Survivors 0.000105 

 FA 5 μM 20 1 × 10

5 Survivors 0.000203 

 FA 7.5 μM 27 1 × 10

5 Survivors 0.000274 
C-60 DRAFT – DO NOT CITE OR QUO

Table C-9. In vitro mammalian mutagenicity: dose response data 

Record 

number Reference PAH Dose Dose units Mutants In number Units % response Comments 

 10 μM 32 1 × 10

5 Survivors 0.000318 

21900 Wangenheim and 

Bolcsfoldi, 1988 

Control 0 mol/L 61 1 × 10

6 Survivors 0.000061 

 Control 0 mol/L 62 1 × 10

6 Survivors 0.000062 Used average of contr

 Average 0 mol/L 62 1 × 10

6 Survivors 0.000062 

 BaP 0.000001 mol/L 65 1 × 10

6 Survivors 0.000065 

 BaP 0.000005 mol/L 243 1 × 10

6 Survivors 0.000243 

 BaP 0.000010 mol/L 858 1 × 10

6 Survivors 0.00086 

 Control 0 mol/L 68 1 × 10

6 Survivors 0.00007 

 FE 0.0000195 mol/L 92 1 × 10

6 Survivors 0.00009 

 FE 0.0000389 mol/L 91 1 × 10

6 Survivors 0.00009 

 FE 0.0000681 mol/L 114 1 × 10

6 Survivors 0.00011 

 FE 0.000122 mol/L 154 1 × 10

6 Survivors 0.00015 

 FE 0.000170 mol/L 147 1 × 10

6 Survivors 0.00015 

 Control 0 mol/L 125 1 × 10

6 Survivors 0.00013 

 Control 0 mol/L 106 1 × 10

6 Survivors 0.00011 

 Average 0 mol/L 116 1 × 10

6 Survivors 0.00012 

 Pyr 0.0000101 mol/L 162 1 × 10

6 Survivors 0.00016 

 Pyr 0.0000151 mol/L 228 1 × 10

6 Survivors 0.00023 

 Pyr 0.0000202 mol/L 345 1 × 10

6 Survivors 0.00035 

 Pyr 0.0000252 mol/L 418 1 × 10

6 Survivors 0.00042 

 Pyr 0.0000302 mol/L 650 1 × 10

6 Survivors 0.00065 

1 
C-61 DRAFT – DO NOT CITE OR QUO

Table C-10. In vitro malignant/morphological cell transformation: data use 

Record 

number Reference Page 

Table 

number

Figure 

number PAHs Data to be extracted 

Basis for 

RPF Comment Notes: 

17610 Casto, 1979 54 I and IV BaP, DBahA TF in number foci per 10

5 surviving 

cells and dose (μg/mL) 

Ratio of slopes 

Data on enhancement of viral transformation not used; no straightforward way to model doseresponse 

Model as incidence data using multistage 

17970 DiPaolo et al., 

1969 

871 3 BaP, DBahA, 

BaA, BeP, DBacA 

Total transformants, total no. colonies, and dose (μg/mL) 

Point estimate 

 Do not use % transforman

appears to be error for DBahA 

18020 Dunkel et al., 

1981 

 Use data as reported in 23720 Pienta 

1977; report under that record 

18080 Emura et al., 

1980 

153, 

154 

I and II BaP, BbF, 

BaA, IP 

T, number of transformed colonies/1,000 survivals in 10 dishes and dose (μg/mL) 

Ratio of slopes 

 Model as incidence data 

using multistage 

14130 Greb et al., 

1980 

147 1 BaP, CH, BaA, 

BbF, DBahA, BeP 

Relative transformation rate (potency) in %/mmol 

Ratio of slopes 

 Relative transformation 

potency at LC50. Slope 

already calculated. 

14640 Krolewski et 

al., 1986 

1,648 1 BaP, CPcdP Transformation frequency per viable 

cell × 10-3; single dose (5 μM) 

Point estimate 

 Use only BaP and CPcdP 

alone (not with IVA/AIA) 

14700 Laaksonen et 

al., 1983 

62 4 BaP, BaA Transformation frequency (no. 

foci/10

5 surviving cells) and dose 

(μM) 

Ratio of slopes 

 Inverse dose response 

relationship poss. due to cytotoxicity. Use peak. 

14850 Lubet et al., 

1983 

992 1 BaP, BeP DwT-III/td (dishes with Type III foci/ 

total dishes) and dose (μg/mL) 

Ratio of slopes 

 Control data in caption (no 

transformants). Model as incidence data. 

24710 Mohapatra et 

al., 1987 

327 1 BaP, BeAC, 

BjAC, BlAC 

Number of dishes scored and percent 

of dishes with Type II or Type III foci and dose (μg/mL) 

Ratio of slope to 

BaP point estimate 

Use BaP incidence as BMR 

Convert percent into numbof dishes and model as incidence data. 

24700 Nesnow et al., 

1990 

224 1 BaP, BlAC Anchorage independent 

colonies/50,000 cells and dose (μg/mL) 

Ratio of slopes 

 Continuous data, no SD fo

controls; use peak. 

7980 Nesnow et al., 

1997 

1,975 I BaP, DBalP Type II and III foci/dish (mean and 

SD) and dose (μM) 

Ratio of slopes 

 Model as continuous data 

7990 Nesnow et al., 

1994 

2,227 I BaP, DBahA Type II and III Foci/dish and dose. 

Use 1 μg/mL dose for DBahA and mean foci/dish (in parentheses). Single dose for BaP 

Point estimate 
C-62 DRAFT – DO NOT CITE OR QUO

Table C-10. In vitro malignant/morphological cell transformation: data use 

Record 

number Reference Page 

Table 

number

Figure 

number PAHs Data to be extracted 

Basis for 

RPF Comment Notes: 

8000 Nesnow et al., 

1993a 

28 I DBkmnoAPH Peak of Type II and III foci/dish. Use 

5 μg/mL dose for DBkmnoAPH and 3 μg/mL dose for BaP. Average number foci/ dish across the two experiments 

Point estimate 

 Peak transformation for ea

compound. DBkmnoAPH reported in paper as CP(3,4)B[a]P 

23720 Pienta et al., 

1977 

648 IV BaP, BaA, 

DBahA 

Transformed colonies/surviving colonies and dose (μg/mL, in row across) 

Ratio of slopes 

 Model as incidence data 

using multistage 

1 
C-63 DRAFT – DO NOT CITE OR QUO

Table C-11. In vitro malignant/morphological cell transformation: dose response data 

Transformation measure 

Record 

number Reference PAH Dose

Dose 

units Mean 

Standard 

deviation

Standard 

error Units n units % Response Notes 

17610 Casto, 1979 Control 0 μg/mL 0 Foci 100,000 Surviving cells 0  BaP 0.62 μg/mL 8 Foci 100,000 Surviving cells 0.00008  BaP 1.25 μg/mL 10 Foci 100,000 Surviving cells 0.0001  DBahA 1.2 μg/mL 0.5 Foci 100,000 Surviving cells 0.000005  DBahA 2.5 μg/mL 1 Foci 100,000 Surviving cells 0.00001 17970 DiPaolo et al., 

1969 

Control 0 μg/mL 0 Transformants 354 No. surviving 0 

 BaP 10 μg/mL 8 Transformants 138 No. surviving 0.058  DBahA 10 μg/mL 11 Transformants 354 No. surviving 0.031  BaA 10 μg/mL 2 Transformants 190 No. surviving 0.011  BeP 10 μg/mL 1 Transformants 172 No. surviving 0.0058  DBacA 10 μg/mL 2 Transformants 181 No. surviving 0.011 18080 Emura et al., 

1980 

Control 0 μg/mL 0 Transformed 

colonies 

1,000 Survivals 0 

 Expt 1 BaP 0.01 μg/mL 0 Transformed 

colonies 

1,000 Survivals 0 

 BaP 0.05 μg/mL 1.1 Transformed 

colonies 

1,000 Survivals 0.0011 

 BaP 0.1 μg/mL 2.9 Transformed 

colonies 

1,000 Survivals 0.0029 

 BaP 0.25 μg/mL 5.3 Transformed 

colonies 

1,000 Survivals 0.0053 

 BaP 0.5 μg/mL 6.8 Transformed 

colonies 

1,000 Survivals 0.0068 

 BbF 0.025 μg/mL 0 Transformed 

colonies 

1,000 Survivals 0 

 BbF 0.1 μg/mL 0.4 Transformed 

colonies 

1,000 Survivals 0.00040 

 BbF 0.25 μg/mL 0.3 Transformed 

colonies 

1,000 Survivals 0.00030 

 BbF 0.5 μg/mL 0.6 Transformed 

colonies 

1,000 Survivals 0.00060 

 BbF 1 μg/mL 1.2 Transformed 

colonies 

1,000 Survivals 0.0012 
C-64 DRAFT – DO NOT CITE OR QUO

Table C-11. In vitro malignant/morphological cell transformation: dose response data 

Transformation measure 

Record 

number Reference PAH Dose

Dose 

units Mean 

Standard 

deviation

Standard 

error Units n units % Response Notes 

 BaA 0.025 μg/mL 0 Transformed 

colonies 

1,000 Survivals 0 

 BaA 0.1 μg/mL 0.3 Transformed 

colonies 

1,000 Survivals 0.00030 

 BaA 0.25 μg/mL 0.3 Transformed 

colonies 

1,000 Survivals 0.00030 

 BaA 0.5 μg/mL 0.6 Transformed 

colonies 

1,000 Survivals 0.00060 

 BaA 1 μg/mL 1 Transformed 

colonies 

1,000 Survivals 0.0010 

 Expt 2 Control 0 μg/mL 0 Transformed 

colonies 

1,000 Survivals 0 

 BaP 0.01 μg/mL 0.4 Transformed 

colonies 

1,000 Survivals 0.00040 

 BaP 0.05 μg/mL 1 Transformed 

colonies 

1,000 Survivals 0.0010 

 BaP 0.1 μg/mL 2.9 Transformed 

colonies 

1,000 Survivals 0.0029 

 BaP 0.25 μg/mL 4.6 Transformed 

colonies 

1,000 Survivals 0.0046 

 BaP 0.5 μg/mL 7.8 Transformed 

colonies 

1,000 Survivals 0.0078 

 IP 0.025 μg/mL 0 Transformed 

colonies 

1,000 Survivals 0 

 IP 0.1 μg/mL 0.3 Transformed 

colonies 

1,000 Survivals 0.00030 

 IP 0.25 μg/mL 0.3 Transformed 

colonies 

1,000 Survivals 0.00030 

 IP 0.5 μg/mL 0.7 Transformed 

colonies 

1,000 Survivals 0.00070 

 IP 1 μg/mL 1 Transformed 

colonies 

1,000 Survivals 0.0010 

14130 Greb et al., 1980 BaP NA 277 %/mmol  CH NA 37 %/mmol  BaA NA 13.9 %/mmol 
C-65 DRAFT – DO NOT CITE OR QUO

Table C-11. In vitro malignant/morphological cell transformation: dose response data 

Transformation measure 

Record 

number Reference PAH Dose

Dose 

units Mean 

Standard 

deviation

Standard 

error Units n units % Response Notes 

 BbF NA 11.5 %/mmol  DBahA NA 0.3 %/mmol  BeP NA 3.1 %/mmol 14640 Krolewski et al., 

1986 

Control 0 μM 0 Transformation 

frequency 

1,000 Viable cells 0 

 BaP 5 μM 5.5 0.7 Transformation 

frequency 

1,000 Viable cells 0.0055 

 CPcdP 5 μM 1.7 0.3 Transformation 

frequency 

1,000 Viable cells 0.0017 

14700 Laaksonen et al., 

1983 

Control 0 μM 0 Foci 1 × 10

5 Surviving cells 0 

 BaP 5 μM 0.8 Foci 1 × 10

5 Surviving cells 0.0000080 Inverse dose response 

relationship poss. due cytotoxicity. Use pea

 BaP 10 μM 0.9 Foci 1 × 10

5 Surviving cells 0.0000090 

 BaP 20 μM 0.3 Foci 1 × 10

5 Surviving cells 0.0000030 

 BaP 40 μM 0.4 Foci 1 × 10

5 Surviving cells 0.0000040 

 Control 0 0 Foci 1 × 10

5 Surviving cells 0 

 BaA 11 μM 1.8 Foci 1 × 10

5 Surviving cells 0.000018 Inverse dose response 

relationship poss. due cytotoxicity. Use pea

 BaA 22 μM 1.5 Foci 1 × 10

5 Surviving cells 0.000015 

 BaA 44 μM 1.1 Foci 1 × 10

5 Surviving cells 0.000011 

 BaA 88 μM 0.8 Foci 1 × 10

5 Surviving cells 0.0000080 

14850 Lubet et al., 

1983 

Control 0 μg/mL 0 Dishes with 

Type III foci 

 Total dishes 0 

 BaP 1 μg/mL 1 Dishes with 

Type III foci 

15 Total dishes 0.067 

 BaP 3 μg/mL 4 Dishes with 

Type III foci 

15 Total dishes 0.267 

 BaP 10 μg/mL 5 Dishes with 

Type III foci 

15 Total dishes 0.333 

 BeP 10 μg/mL 0 Dishes with 

Type III foci 

15 Total dishes 0 
C-66 DRAFT – DO NOT CITE OR QUO

Table C-11. In vitro malignant/morphological cell transformation: dose response data 

Transformation measure 

Record 

number Reference PAH Dose

Dose 

units Mean 

Standard 

deviation

Standard 

error Units n units % Response Notes 

 BeP 30 μg/mL 1 Dishes with 

Type III foci 

15 Total dishes 0.067 

 BeP 100 μg/mL 7 Dishes with 

Type III foci 

15 Total dishes 0.467 

24710 Mohapatra et al., 

1987 

Control 0 μg/mL 0 Dishes with 

Type II or III foci 

48 Dishes scored 0 

 BaP 1 μg/mL 44 Dishes with 

Type II or III foci 

48 Dishes scored 0.92 

 BjAC 0.01 μg/mL 2 Dishes with 

Type II or III foci 

48 Dishes scored 0.04 

 BjAC 0.05 μg/mL 5 Dishes with 

Type II or III foci 

48 Dishes scored 0.1 

 BjAC 0.5 μg/mL 34 Dishes with 

Type II or III foci 

48 Dishes scored 0.71 

 BjAC 1 μg/mL 45 Dishes with 

Type II or III foci 

48 Dishes scored 0.94 

 BjAC 2 μg/mL 48 Dishes with 

Type II or III foci 

48 Dishes scored 1 

 Control 0 μg/mL 0 Dishes with 

Type II or III foci 

60 Dishes scored 0 

 BaP 1 μg/mL 50 Dishes with 

Type II or III foci 

60 Dishes scored 0.83 

 BlAC 0.5 μg/mL 8 Dishes with 

Type II or III foci 

60 Dishes scored 0.13 

 BlAC 1 μg/mL 14 Dishes with 

Type II or III foci 

60 Dishes scored 0.26 

 BlAC 2.5 μg/mL 31 Dishes with 

Type II or III foci 

60 Dishes scored 0.52 

 BlAC 5 μg/mL 42 Dishes with 

Type II or III foci 

60 Dishes scored 0.7 

 BlAC 10 μg/mL 51 Dishes with 

Type II or III foci 

60 Dishes scored 0.85 

 Control 0 μg/mL 0 Dishes with 

Type II or III foci 

36 Dishes scored 0 

 BaP 1 μg/mL 31 Dishes with 

Type II or III foci 

36 Dishes scored 0.86 
C-67 DRAFT – DO NOT CITE OR QUO

Table C-11. In vitro malignant/morphological cell transformation: dose response data 

Transformation measure 

Record 

number Reference PAH Dose

Dose 

units Mean 

Standard 

deviation

Standard 

error Units n units % Response Notes 

 BeAC 0.5 μg/mL 4 Dishes with 

Type II or III foci 

36 Dishes scored 0.11 

 BeAC 1 μg/mL 6 Dishes with 

Type II or III foci 

36 Dishes scored 0.17 

 BeAC 2.5 μg/mL 13 Dishes with 

Type II or III foci 

36 Dishes scored 0.36 

 BeAC 5 μg/mL 15 Dishes with 

Type II or III foci 

36 Dishes scored 0.42 

 BeAC 10 μg/mL 21 Dishes with 

Type II or III foci 

36 Dishes scored 0.58 

24700 Nesnow et al., 

1990 

Acetone 0 μg/mL 25 Anchorage 

independent colonies/50,000 cells 

 BaP 0.1 μg/mL 43 14.7 Anchorage 

independent colonies/50,000 cells 

 BaP 0.5 μg/mL 42 20.7 Anchorage 

independent colonies/50,000 cells 

 BaP 2.5 μg/mL 39 19.5 Anchorage 

independent colonies/50,000 cells 

 BaP 10 μg/mL 72 23.1 Anchorage 

independent colonies/50,000 cells 

 Acetone 0 μg/mL 30 Anchorage 

independent colonies/50,000 cells 

 BlAC 0.1 μg/mL 74 5.2 Anchorage 

independent colonies/50,000 cells 
C-68 DRAFT – DO NOT CITE OR QUO

Table C-11. In vitro malignant/morphological cell transformation: dose response data 

Transformation measure 

Record 

number Reference PAH Dose

Dose 

units Mean 

Standard 

deviation

Standard 

error Units n units % Response Notes 

 BlAC 0.5 μg/mL 68 14.4 Anchorage 

independent colonies/50,000 cells 

 BlAC 2.5 μg/mL 123 15.6 Anchorage 

independent colonies/50,000 cells 

 BlAC 10 μg/mL 150 16.8 Anchorage 

independent colonies/50,000 cells 

7980 Nesnow et al., 

1997 

Control 0 μM 0 0 Type II and III 

foci/dish 

 BaP 0.4 μM 0.44 0.24 Type II and III 

foci/dish 

 BaP 1.2 μM 1.25 0.15 Type II and III 

foci/dish 

 BaP 4 μM 2.54 0.56 Type II and III 

foci/dish 

 DBalP 0.003

3 

μM 0.14 0.35 Type II and III 

foci/dish 

 DBalP 0.1 μM 1 0.24 Type II and III 

foci/dish 

 DBalP 0.33 μM 1.74 0.78 Type II and III 

foci/dish 

7990 Nesnow et al., 

1994 

Control 0 μg/mL 0.06 0.10 Type II and III 

foci/dish 

 BaP 1 μg/mL 1 0.43 Type II and III 

foci/dish 

 DBahA 0.25 μg/mL 0.23 0.21 Type II and III 

foci/dish 

 DBahA 0.5 μg/mL 0.25 0.33 Type II and III 

foci/dish 

 DBahA 1 μg/mL 0.43 0.11 Type II and III 

foci/dish 
C-69 DRAFT – DO NOT CITE OR QUO

Table C-11. In vitro malignant/morphological cell transformation: dose response data 

Transformation measure 

Record 

number Reference PAH Dose

Dose 

units Mean 

Standard 

deviation

Standard 

error Units n units % Response Notes 

 DBahA 2.5 μg/mL 0.29 0.085 Type II and III 

foci/dish 

8000 Nesnow et al., 

1993a 

Control 0 μg/mL 0 Type II and III 

foci/dish 

 BaP 0.3 μg/mL 0.48 Type II and III 

foci/dish 

 BaP 1 μg/mL 0.665 Type II and III 

foci/dish 

 BaP 3 μg/mL 1.4 Type II and III 

foci/dish 

 Control 0 μg/mL 0 Type II and III 

foci/dish 

 DBkmnoA

PH 

0.5 μg/mL 0.23 Type II and III 

foci/dish 

 DBkmnoA

PH 

1 μg/mL 0.52 Type II and III 

foci/dish 

 DBkmnoA

PH 

2.5 μg/mL 0.605 Type II and III 

foci/dish 

 DBkmnoA

PH 

5 μg/mL 1.085 Type II and III 

foci/dish 

23720 Pienta et al., 

1977 

Control 0 μg/mL 0 Transformed 

colonies 

504 Surviving 

colonies 

0 BaP and BaA data als

reported in 18020 Dun1981 

 BaP 1 μg/mL 1 Transformed 

colonies 

393 Surviving 

colonies 

0.0025 

 BaP 5 μg/mL 2 Transformed 

colonies 

406 Surviving 

colonies 

0.0049 

 BaP 10 μg/mL 3 Transformed 

colonies 

434 Surviving 

colonies 

0.0069 

 BaP 20 μg/mL 5 Transformed 

colonies 

410 Surviving 

colonies 

0.0122 

 BaP 40 μg/mL 4 Transformed 

colonies 

427 Surviving 

colonies 

0.0094 

 Control 0 μg/mL 0 Transformed 

colonies 

229 Surviving 

colonies 

0 
C-70 DRAFT – DO NOT CITE OR QUO

Table C-11. In vitro malignant/morphological cell transformation: dose response data 

Transformation measure 

Record 

number Reference PAH Dose

Dose 

units Mean 

Standard 

deviation

Standard 

error Units n units % Response Notes 

 BaA 0.1 μg/mL 1 Transformed 

colonies 

225 Surviving 

colonies 

0.0044 

 BaA 0.5 μg/mL 2 Transformed 

colonies 

252 Surviving 

colonies 

0.0079 

 BaA 1 μg/mL 2 Transformed 

colonies 

193 Surviving 

colonies 

0.0104 

 BaA 5 μg/mL 1 Transformed 

colonies 

312 Surviving 

colonies 

0.0032 

 BaA 10 μg/mL 7 Transformed 

colonies 

250 Surviving 

colonies 

0.028 

 Control 0 μg/mL 0 Transformed 

colonies 

229 Surviving 

colonies 

0 

 DBahA 0.1 μg/mL 0 Transformed 

colonies 

219 Surviving 

colonies 

0 

 DBahA 0.5 μg/mL 4 Transformed 

colonies 

233 Surviving 

colonies 

0.0172 

 DBahA 1 μg/mL 4 Transformed 

colonies 

217 Surviving 

colonies 

0.0184 

 DBahA 5 μg/mL 5 Transformed 

colonies 

270 Surviving 

colonies 

0.0185 

 DBahA 10 μg/mL 0 Transformed 

colonies 

232 Surviving 

colonies 

0 
C-71 DRAFT – DO NOT CITE OR QUOTE 

Table C-12. In vitro DNA adducts: data use 

Record 

number Reference Page

Table 

number

Figure 

number PAHs 

Data to be 

extracted 

Basis for 

RPF Comment Notes: 

16890 Allen and 

Coombs, 

1980 

245 1 BaP, BaA μmol compound/mol 

DNA P 

Point 

estimate 

Adducts in 

nuclear and 

mitochondrial 

DNA 

Calculate 

separate 

RPFs for 

nuclear and 

mitochondrial DNA 

6300 Binkova et 

al., 2000 

62 3 BaP, 

DBalP 

Adducts at 

each dose 

level 

Ratio of 

slopes 

Slope of adduct 

vs. dose curve 

May need to 

drop high 

dose data 

for adequate 

fit 

9510 Bryla and 

Weyand, 

1992 

39 1 BaP, BaA, 

DBacA 

Adducts at 

each dose 

level 

Ratio of 

slopes 

Slope of adduct 

vs. dose curve 

under light 

conditions 

(max response 

for all 

compounds) 

22800 Grover and 

Sims, 1968

160 1 BaP, 

DBahA, 

DBacA, 

BaA, Pyr, 

PH 

Reaction 

with DNA 

Point 

estimate 

 

10660 Johnsen et 

al., 1998 

80 2 BjAC, 

BlAC, BaP

Total adduct 

levels in 

human 

lymphocytes 

and HL-60 

cells 

Point 

estimate 

Total adducts 

formed in 

human 

lymphocytes or 

HL-60 cells 

Calc RPFs 

separately 

by cell type 

10670 Johnsen et 

al., 1997 

196 II BjAC, 

BlAC, BaP

DNA adduct 

levels in 

PCB-treated 

rat lung cells 

Point 

estimate 

Adducts in 

PCB-treated rat 

lung Clara and 

Type 2 cells 

Calc RPFs 

separately 

by cell type 

7870 MelendezColon et 

al., 2000 

13 2 BaP, 

DBalP 

Stable DNA 

adducts at 

each dose 

level 

Ratio of 

slopes 

Slope of adduct 

vs. dose curve 

at two doses 

21200 Segerback 

and 

Vodicka, 

1993 

2,465 3 Pyr, BghiP, 

FA, 

DBahA, 

BbF, BaP, 

BaA, CH 

Total adduct 

levels 

Point 

estimate 

Total adduct 

level in 

optimized 

nuclease P1 

adduct 

enrichment 

procedure 
C-72 DRAFT – DO NOT CITE OR QUO

Table C-13. In vitro DNA adducts: dose response data 

DNA Adducts 

Record 

number Reference PAH Dose 

Dose 

units Mean 

Standard 

deviation Adduct units n Units Notes 

16890 Allen and 

Coombs, 1980 

BaP 0.235 μg/mL 7.5 1.9 μmol/mol DNA 

P 

 Nuclear DNA 

 BaA 0.644 μg/mL 0.44 0.11 μmol/mol DNA 

P 

 Nuclear DNA 

 BaP 0.235 μg/mL 413 164 μmol/mol DNA 

P 

 Mitochondrial DNA 

 BaA 0.644 μg/mL 104 40.2 μmol/mol DNA 

P 

 Mitochondrial DNA 

6300 Binkova et 

al., 2000 

BaP 0.010 μM 1.8 1.16 Adducts 1 × 10

8 Nucleotides 

 0.10 μM 18 7.18 Adducts 1 × 10

8 Nucleotides 

 0.40 μM 95 39.4 Adducts 1 × 10

8 Nucleotides 

 1.0 μM 258 115 Adducts 1 × 10

8 Nucleotides 

 4.0 μM 205 81.9 Adducts 1 × 10

8 Nucleotides 

 10 μM 69 21.9 Adducts 1 × 10

8 Nucleotides 

 40 μM 37 10.8 Adducts 1 × 10

8 Nucleotides 

 DBalP 0.010 μM 179 55.3 Adducts 1 × 10

8 Nucleotides 

 0.020 μM 534 52.6 Adducts 1 × 10

8 Nucleotides 

 0.040 μM 1,304 375 Adducts 1 × 10

8 Nucleotides 

 0.080 μM 1,696 644 Adducts 1 × 10

8 Nucleotides 

 0.10 μM 2,317 774 Adducts 1 × 10

8 Nucleotides 

 0.40 μM 1,971 729 Adducts 1 × 10

8 Nucleotides 

 1.0 μM 632 170 Adducts 1 × 10

8 Nucleotides 

9510 Bryla and 

Weyand, 1992 

BaP 0.12 nmol 0.17 Adducts 1 × 10

7 Nucleotides Light conditions; max for BaP and others 

 BaP 12 nmol 1.37 Adducts 1 × 10

7 Nucleotides 

 BaP 120 nmol 2.21 Adducts 1 × 10

7 Nucleotides 

 BaP 600 nmol 5.45 Adducts 1 × 10

7 Nucleotides 

 BaA 0.12 nmol 0.15 Adducts 1 × 10

7 Nucleotides 

 BaA 12 nmol 0.09 Adducts 1 × 10

7 Nucleotides 

 BaA 120 nmol 0.8 Adducts 1 × 10

7 Nucleotides 
C-73 DRAFT – DO NOT CITE OR QUO

Table C-13. In vitro DNA adducts: dose response data 

DNA Adducts 

Record 

number Reference PAH Dose 

Dose 

units Mean 

Standard 

deviation Adduct units n Units Notes 

 BaA 600 nmol 0.95 Adducts 1 × 10

7 Nucleotides 

 DBacA 0.12 nmol 0 Adducts 1 × 10

7 Nucleotides 

 DBacA 12 nmol 0.06 Adducts 1 × 10

7 Nucleotides 

 DBacA 120 nmol 0.57 Adducts 1 × 10

7 Nucleotides 

 DBacA 600 nmol 1.76 Adducts 1 × 10

7 Nucleotides 

22800 Grover and 

Sims, 1968 

BaP 5 μg 1.41 μmol/g-atom of 

DNA P 

 DBahA 5 μg 0.44 μmol/g-atom of 

DNA P 

 DBacA 5 μg 0.56 μmol/g-atom of 

DNA P 

 BaA 5 μg 0.7 μmol/g-atom of 

DNA P 

 Pyr 5 μg 0.31 μmol/g-atom of 

DNA P 

 PH 5 μg 0.05 μmol/g-atom of 

DNA P 

10670 Johnsen et 

al., 1997 

BaP 30 μg/mL 0.05 fmol adducts/μg 

DNA 

 Clara cells 

 BjAC 30 μg/mL 0.15 fmol adducts/μg 

DNA 

 Clara cells 

 BlAC 30 μg/mL 0.24 fmol adducts/μg 

DNA 

 Clara cells 

 BaP 30 μg/mL 0.02 fmol adducts/μg 

DNA 

 Type 2 cells 

 BjAC 30 μg/mL 0.06 fmol adducts/μg 

DNA 

 Type 2 cells 

 BlAC 30 μg/mL 0.03 fmol adducts/μg 

DNA 

 Type 2 cells 

10660 Johnsen et 

al., 1998 

BaP 30 μg/mL 0.333 0.093 fmol adducts/μg 

DNA 

3 Human lymphocytes 

 BjAC 30 μg/mL 0.110 0.026 fmol adducts/μg 

DNA 

3 Human lymphocytes 

 BlAC 30 μg/mL 1.089 0.595 fmol adducts/μg 

DNA 

3 Human Lymphocytes 
C-74 DRAFT – DO NOT CITE OR QUO

Table C-13. In vitro DNA adducts: dose response data 

DNA Adducts 

Record 

number Reference PAH Dose 

Dose 

units Mean 

Standard 

deviation Adduct units n Units Notes 

 BaP 30 μg/mL 0.239 0.172 fmol adducts/μg 

DNA 

3 HL-60 Cells 

 BjAC 30 μg/mL 0.149 0.146 fmol adducts/μg 

DNA 

3 HL-60 Cells 

 BlAC 30 μg/mL 0.942 0.344 fmol adducts/μg 

DNA 

3 HL-60 Cells 

7870 MelendezColon et al., 

2000 

BaP 1 μm 18 8.07 Stable adducts 1 × 10

6 Nucleotides 

 BaP 2 μm 34 6.46 Stable adducts 1 × 10

6 Nucleotides 

 DBalP 1 μm 254 4.30 Stable adducts 1 × 10

6 Nucleotides 

 DBalP 2 μm 348 17.20 Stable adducts 1 × 10

6 Nucleotides 

21200 Segerback 

and Vodicka, 1993 

BaP 100 mM 15 μmol adducts per 

mol dNp 

 Pyr 100 mM 0.14 μmol adducts per 

mol dNp 

 BghiP 100 mM 0.50 μmol adducts per 

mol dNp 

 FA 100 mM 1.5 μmol adducts per 

mol dNp 

 DBahA 100 mM 2.8 μmol adducts per 

mol dNp 

 BbF 100 mM 3.7 μmol adducts per 

mol dNp 

 BaA 100 mM 30 μmol adducts per 

mol dNp 

 CH 100 mM 50 μmol adducts per 

mol dNp 

1 2 
C-75 DRAFT – DO NOT CITE OR QUO

Table C-14. In vitro DNA damage: data use 

Record 

number Reference Page 

Table 

number

Figure 

number PAHs Data to be extracted Basis for RPF Comment Notes

16840 Agrelo and 

Amos, 1981 

531 2 BaP, Pyr Hydroxyurea inhibited 

3H Thymidine 

incorporation into cells (dpm) and dose 

(μg/mL). Use 10 μg/mL dose for BaP and 100 μg/mL dose for pyrene. 

Point estimate 

23790 Ichinotsubo 

et al., 1977

56 Table II BaP, DBaiP, DBahA Use column designated JC5519 +S9 

for BaP, DBaiP, and DBahA; dose as μg/well and response as diameter of zone of inhibition (mm). The control is wild type strain AB1157

Point estimate E. coli Rec BC, S9 

identification unknown

10660 Johnsen et 

al., 1998 

82 4 BaP, BjAC, BlAC DNA damage (NAAC, 10-3

h-1), std dev

and dose (μg/mL) for both human lymphocytes and HL-60 cells. Use 24 h + 1 h AraC/HU data (crosshatched bars) 

Ratio of slopes (human lymphocytes); point estimates (HL-60 cells) 

 Model as 

continuoudata. 

19740 Martin et al., 

1978 

2,624 1 BaP, BeP, BaA, 

DBacA, DBahA 

Maximum dpm/μg DNA above 

background and dose (M). Dose is in 

column marked "M". 

Point estimate Background already 

subtracted 

19830 MerschSundermann 

et al., 1992 

3-6 2 BaP, AA, BaA, BbF, 

BghiF, BjF, BbFE, BghiP, BeP, CH, DBacA, DBahA, DBalP, DBahP, DBaiP, FA, IP, PH, Tphen 

SOS induction potential (SOSIP) for assay (+S9) for each compound (already incorporates dose) 

Ratio of SOSIPs SOSIP reported in text 

as slope of steepest 

portion of the 

induction factor (IF) dose-response curve. 

No mode

necessaryslopes reported itext. 

20810 Robinson 

and Mitchell, 1981 

520 1 BaP, Pyr Maximum 

3H-TDR incorporation and 

dose (test concentration in μg/mL in 

parentheses after maximum) for rows with 

metabolic activation (+). Use compoundspecific background 

3H -TDR 

incorporation in same row. 

Point estimate 

20940 Rossman et 

al., 1991 

354 2 BaP, AC, DBacA, 

DBahA, PH 

Max enhancement of prophage induction over background and dose (amount at 

max, in μg/well) for those rows with S9 (+ rows). 

Point estimate Background already 

addressed 

21730 Tong et al., 

1981b 

480 I BaP, BaA DNA repair grains/nucleus, std dev, and 

dose (M). Four doses BaA, three doses BaP and DMSO control. 

Ratio of slopes Model as 

continuoudata. 

1 
C-76 DRAFT – DO NOT CITE OR QUO

1 

Table C-15. In vitro DNA damage: dose response data 

Record DNA Damage 

number Reference PAH Dose 

Dose 

units Endpoint Mean SD Units n Notes 

16840 Agrelo and 

Amos, 1981 

Control 0 μg/mL Unscheduled DNA synthesis 177 dpm HU inhibited 

 BaP 0.001 μg/mL Unscheduled DNA synthesis 195 dpm HU inhibited  BaP 0.01 μg/mL Unscheduled DNA synthesis 126 dpm HU inhibited  BaP 0.1 μg/mL Unscheduled DNA synthesis 262 dpm HU inhibited  BaP 1 μg/mL Unscheduled DNA synthesis 818 dpm HU inhibited  BaP 10 μg/mL Unscheduled DNA synthesis 2,270 dpm HU inhibited  BaP 100 μg/mL Unscheduled DNA synthesis 819 dpm HU inhibited  BaP 1,000 μg/mL Unscheduled DNA synthesis 373 dpm HU inhibited  Control 0 μg/mL Unscheduled DNA synthesis 1,168 dpm HU inhibited  Pyr 0.032 μg/mL Unscheduled DNA synthesis 1,293 dpm HU inhibited  Pyr 0.16 μg/mL Unscheduled DNA synthesis 1,192 dpm HU inhibited  Pyr 0.8 μg/mL Unscheduled DNA synthesis 1,367 dpm HU inhibited  Pyr 4 μg/mL Unscheduled DNA synthesis 1,510 dpm HU inhibited  Pyr 20 μg/mL Unscheduled DNA synthesis 1,694 dpm HU inhibited  Pyr 100 μg/mL Unscheduled DNA synthesis 1,716 dpm HU inhibited 23790 Ichinotsubo et al., 

1977

Control 0 DNA damage 0 Diameter of zone 

of inhibition mm

BaP 70 μg/well DNA damage 6 Diameter of zone 

of inhibition mm

Control 0 DNA damage 0 Diameter of zone 

of inhibition mm

DBaiP 600 μg/well DNA damage 10 Diameter of zone 

of inhibition mm

Control 0 DNA damage 0 Diameter of zone 

of inhibition mm

DBahA 25 μg/well DNA damage 10 Diameter of zone 

of inhibition mm

10660 Johnsen et al., 

1998 

DMSO 0 μg/mL DNA damage 4.4 1.3 NAAC, 10-3 h-1 3 Human lymphocytes with 

AraC/HU 
C-77 DRAFT – DO NOT CITE OR QUO

Table C-15. In vitro DNA damage: dose response data 

Record DNA Damage 

number Reference PAH Dose 

Dose 

units Endpoint Mean SD Units n Notes 

 BaP 3 μg/mL DNA damage 12 3.2 NAAC, 10-3 h-6 3 Human lymphocytes with 

AraC/HU. No continuous lineamodel fit 

 30 μg/mL DNA damage 15 2.7 NAAC, 10-3 h-7 3 Human lymphocytes with 

AraC/HU 

 BjAC 3 μg/mL DNA damage 6.0 2.1 NAAC, 10-3 h-2 3 Human lymphocytes with 

AraC/HU 

 30 μg/mL DNA damage 9.4 3.4 NAAC, 10-3 h-3 3 Human lymphocytes with 

AraC/HU 

 BIAC 3 μg/mL DNA damage 8.2 3.2 NAAC, 10-3 h-4 3 Human lymphocytes with 

AraC/HU. No continuous lineamodel fit 

 30 μg/mL DNA damage 9.3 2.1 NAAC, 10-3 h-5 3 Human lymphocytes with 

AraC/HU 

 DMSO 0 μg/mL DNA damage 7.8 3.1 NAAC, 10-3 h-5 3 HL-60 cells with AraC/HU  BaP 30 μg/mL DNA damage 13.2 9.5 NAAC, 10-3 h-5 3 HL-60 cells with AraC/HU  BjAC 30 μg/mL DNA damage 9.6 3.0 NAAC, 10-3 h-5 3 HL-60 cells with AraC/HU  BIAC 30 μg/mL DNA damage 11.6 5.5 NAAC, 10-3 h-5 3 HL-60 cells with AraC/HU 19740 Martin et al., 

1978 

BaP 1 × 10-5 M Unscheduled DNA synthesis 210 Max dpm/μg 

DNA 

 Increase above background 

 BeP 1 × 10-6 M Unscheduled DNA synthesis 256 Max dpm/μg 

DNA 

 Increase above background 

 BaA 1 × 10-7 M Unscheduled DNA synthesis 59 Max dpm/μg 

DNA 

 Increase above background 

 DBacA 1 × 10-5 M Unscheduled DNA synthesis 97 Max dpm/μg 

DNA 

 Increase above background 

 DBahA 1 × 10-5 M Unscheduled DNA synthesis 96 Max dpm/μg 

DNA 

 Increase above background 

19830 MerschSundermann et 

al., 1992 

BaP NA SOS induction potential 0.605 NA Steepest slope of induction fact

dose-response curve; + S9 

 AA NA SOS induction potential 0.142 NA Steepest slope of induction fact

dose-response curve; + S9 
C-78 DRAFT – DO NOT CITE OR QUO

Table C-15. In vitro DNA damage: dose response data 

Record DNA Damage 

number Reference PAH Dose 

Dose 

units Endpoint Mean SD Units n Notes 

 BaA NA SOS induction potential 0.1 NA Steepest slope of induction fact

dose-response curve; + S9 

 BbF NA SOS induction potential 0.045 NA Steepest slope of induction fact

dose-response curve; + S9 

 BghiF NA SOS induction potential 0.34 NA Steepest slope of induction fact

dose-response curve; + S9 

 BjF NA SOS induction potential 0.254 NA Steepest slope of induction fact

dose-response curve; + S9 

 BbFE NA SOS induction potential 0.024 NA Steepest slope of induction fact

dose-response curve; + S9 

 BghiP NA SOS induction potential 0.033 NA Steepest slope of induction fact

dose-response curve; + S9 

 BeP NA SOS induction potential 0.032 NA Steepest slope of induction fact

dose-response curve; + S9 

 CH NA SOS induction potential 0.221 NA Steepest slope of induction fact

dose-response curve; + S9 

 DBacA NA SOS induction potential 0.104 NA Steepest slope of induction fact

dose-response curve; + S9 

 DBahA NA SOS induction potential 0.039 NA Steepest slope of induction fact

dose-response curve; + S9 

 DBalP NA SOS induction potential 2.1 NA Steepest slope of induction fact

dose-response curve; + S9 

 DBahP NA SOS induction potential 0.117 NA Steepest slope of induction fact

dose-response curve; + S9 

 DBaiP NA SOS induction potential 0.174 NA Steepest slope of induction fact

dose-response curve; + S9 

 FA NA SOS induction potential 0.412 NA Steepest slope of induction fact

dose-response curve; + S9 

 IP NA SOS induction potential 0.036 NA Steepest slope of induction fact

dose-response curve; + S9 

 PH NA SOS induction potential 0.053 NA Steepest slope of induction fact

dose-response curve; + S9 
C-79 DRAFT – DO NOT CITE OR QUO

Table C-15. In vitro DNA damage: dose response data 

Record DNA Damage 

number Reference PAH Dose 

Dose 

units Endpoint Mean SD Units n Notes 

 Tphen NA SOS induction potential 0.26 NA Steepest slope of induction fact

dose-response curve; + S9 

20810 Robinson and 

Mitchell, 1981 

Control 0 μg/mL Unscheduled DNA synthesis 53 4 

3H-TdR incorporation 

 Max 3H-TdR incorporation 

 BaP 10 μg/mL Unscheduled DNA synthesis 142 7 

3H-TdR incorporation 

 Max 3H-TdR incorporation 

 Control 0 μg/mL Unscheduled DNA synthesis 52 2 

3H-TdR incorporation 

 Max 3H-TdR incorporation 

 Pyr 7.2 μg/mL Unscheduled DNA synthesis 115 9 

3H-TdR incorporation 

 Max 3H-TdR incorporation 

20940 Rossman et al., 

1991 

BaP 12.5 μg/mL DNA damage 10.4 Lambda prophage induction Max enhancement over background 

 AC 12.5 μg/mL DNA damage 4.8 Lambda prophage induction Max enhancement over background  DBacA 1.44 μg/mL DNA damage 8 Lambda prophage induction Max enhancement over background  DBahA 2 μg/mL DNA damage 4 Lambda prophage induction Max enhancement over background  PH 25 μg/mL DNA damage 4.5 Lambda prophage induction Max enhancement over background 21730 Tong et al., 

1981b 

Control 0 M Unscheduled DNA synthesis 0.1 0.1 Grains/nucleus 

 BaP 1 × 10-4 M Unscheduled DNA synthesis 45.1 3.7 Grains/nucleus  BaP 5 × 10-4 M Unscheduled DNA synthesis 47.7 3.7 Grains/nucleus  BaP 1 × 10-3 M Unscheduled DNA synthesis 65.6 17.8 Grains/nucleus  BaA 5 × 10-5 M Unscheduled DNA synthesis 0.6 Grains/nucleus  BaA 1 × 10-4 M Unscheduled DNA synthesis 14.8 2.6 Grains/nucleus  BaA 5 × 10-4 M Unscheduled DNA synthesis 17.2 6 Grains/nucleus  BaA 1 × 10-3 M Unscheduled DNA synthesis Toxic Grains/nucleus 

1 
C-80 DRAFT – DO NOT CITE OR QUOTE 

Table C-16. In vitro clastogenicity: data use 

Record 

number Reference Page

Table 

number PAHs Data to be used 

Basis for 

RPF Comment 

14620 Kochhar, 1982 846 Not 

numbered

BaP, 

BaA 

% cells with 

aberrations and dose 

(μg/mL) 

Ratio of 

slopes 

Model as incidence 

data. 

14640 Krolewski et al., 

1986 

1,648 II BaP, 

CPcdP

Mean no. SCE/

chromosome, std dev, 

and dose (μM) 

Ratio of 

slopes 

Use first column of 

data; not data with AIA 

or IVA. Model as 

continuous data. 

19690 Mane et al., 1990 81 III BaP, 

BaA 

SCE frequencies/for 

V79 cell + rat MEC 

and dose 

Point 

estimates 

Use SCE data for V79 + 

rat MEC only. 

21710 Tong et al., 

1981a 

469 1 BaP, 

BaA 

SCE/cell, std dev, and 

dose 

Point 

estimates 

Continuous data, no n 

provided in study. 

1 
C-81 DRAFT – DO NOT CITE OR QUOTE 

Table C-17. In vitro clastogenicity: dose response data 

Clastogenicity 

Record 

number Reference PAH Dose 

Dose 

units n Mean 

Standard 

deviation Units Notes 

14620 Kochhar, 

1982 

Control 0 μg/mL 100 0.06 Fraction cells with 

aberrations 

 BaP 0.6 μg/mL 100 0.23 Fraction cells with 

aberrations 

 BaP 1.25 μg/mL 100 0.32 Fraction cells with 

aberrations 

 BaP 2.5 μg/mL 100 0.45 Fraction cells with 

aberrations 

 BaP 5 μg/mL 100 0.56 Fraction cells with 

aberrations 

 BaA 0.6 μg/mL 100 0.17 Fraction cells with 

aberrations 

 BaA 1.25 μg/mL 100 0.23 Fraction cells with 

aberrations 

 BaA 2.5 μg/mL 100 0.3 Fraction cells with 

aberrations 

 BaA 5 μg/mL 100 0.38 Fraction cells with 

aberrations 

14640 Krolewski et 

al., 1986 

Control 0 μM 30 0.147 0.059 SCE/ 

 BaP 1 μM 30 0.874 0.275 SCE/ 

 BaP 5 μM 30 0.932 0.266 SCE/ 

 CPcdP 1 μM 30 0.348 0.119 SCE/ 

 CPcdP 5 μM 30 0.432 0.15 SCE/ 

19690 Mane et al., 

1990 

Control 0 μg/mL 0.3 1 SCE frequency/ For V79 cell + rat 

MEC 

 BaP 1 μg/mL 3 1 SCE frequency/ For V79 cell + rat 

MEC 

 BaA 1 μg/mL 0.7 0.5 SCE frequency/ For V79 cell + rat 

MEC 

21710 Tong et al., 

1981a 

Control 0 M 11.15 3.81 SCE/cell 

 BaP 1 × 10-6 M 16.15 3.83 SCE/cell 

 BaP 1 × 10-5 M 59.75 16.96 SCE/cell 

 BaP 1 × 10-4 M 103.3 22.75 SCE/cell 

 Control 0 M 15.75 5.18 SCE/cell 

 BaA 1 × 10-5 M 21.2 9.59 SCE/cell 

 BaA 1 × 10-4 M 29.15 9.93 SCE/cell 

 BaA 1 × 10-3 M 26.2 6.96 SCE/cell 
C-82 DRAFT – DO NOT CITE OR QUO

Table C-18. In vivo DNA adducts: data use 

Record 

number Reference Page

Table 

number

Figure 

number PAHs 

Data to be 

extracted 

Basis for 

RPF Comment Notes 

6210 Arif et al., 

1997 

36 4 DBalP and BaP Mean adduct levels 

for heart, pancreas, 

bladder, liver 

Point estimate 

Mean adduct levels summed across mammary epithelial, lung, heart, 

pancreas, bladder, liver 

17630 Cavalieri et 

al., 1981a 

491 3 CPcdP, ACEP 

(reported in paper as CPAP), BaP 

Done Point 

estimate 

DNA-bound PAH in mouse skin after 4 hr or 24 hr treatment 

Calculate separatRPFs for 4 hr and 24 hr treatment 

18810 Hughes 

and Phillips, 1990 

1,61

4 

 3 DBalP, DBaeP, 

DBahP, DBaiP, BaP 

AUC for skin and lung through 84 d 

Point estimate 

Sum of AUCs for skin and lung 0–84 d 

11190 Mass et al., 

1993 

188 1 BjAC, BaP Done Ratio of 

Slopes 

AUC (adduct-time curve) vs. dose for lung adducts 24–72 hr 

8010 Nesnow et 

al., 1993b 

39 1 and 2 BbF, BaP AUC for lung, liver, 

and PBL through 56 d 

Point estimate 

Sum of AUCs for lung, liver, and lymphocytes 0–56 d 

24590/ 20920 

Nesnow et 

al., 1998b; Ross et al., 1995 

402 2 BaP, BbF, 

DBahA, CPcdP, DBalP 

Done Ratio of 

Slopes 

Slope of TIDAL/dose (slope reported in 24590 based on data from 20920). DBalP data reported in separate study w/o BaP concurrent 

22810 Phillips et 

al., 1979 

205 I DBahA, DBacA, 

BaP 

Done Point 

estimate 

Peak binding in mouse skin. BaA dropped; not clear if reported level is 

peak. 

24790 Kligerman 

et al., 2002

846 1 BaA, BaP, BbF, 

CH 

Done Point 

estimate 

Adducts in mouse or rat PBLs at single time point after either intraperitoneal or gavage administration 

Calculate separatRPFs for intraperitoneal angavage, rat and mouse 

1 2 
C-83 DRAFT – DO NOT CITE OR QUO

Table C-19. In vivo DNA adducts: dose response data 

DNA adducts 

Record 

number Reference PAH Species Dose Dose units Organ Time Mean 

Standard 

deviation 

Standard 

error Adduct units 

Slope of 

AUC vs. 

dose Comments 

6210 Arif et al., 

1997 

Control Rat 0 μmol/ mammary gland Liver 0 Adducts/10

9

nucleotides 

 BaP Rat 0.25 μmol/mammary gland Mammary gland 300 45 Adducts/10

9

nucleotides 

 BaP Rat 0.25 μmol/mammary gland Lung 11 1.3 Adducts/10

9

nucleotides 

 BaP Rat 0.25 μmol/mammary gland Heart 9.5 Adducts/10

9

nucleotides 

 BaP Rat 0.25 μmol/mammary gland Pancreas 0 Adducts/10

9

nucleotides 

 BaP Rat 0.25 μmol/mammary gland Bladder 0 Adducts/10

9

nucleotides 

 BaP Rat 0.25 μmol/mammary gland Liver 4.5 Adducts/10

9

nucleotides 

 Sum 324.74  DBalP Rat 0.25 μmol/mammary gland Mammary gland 1,878 378 Adducts/10

9

nucleotides 

 DBalP Rat 0.25 μmol/mammary gland Lung 85 24 Adducts/10

9

nucleotides 

 DBalP Rat 0.25 μmol/mammary gland Heart 64 Adducts/10

9

nucleotides 

 DBalP Rat 0.25 μmol/mammary gland Pancreas 32 Adducts/10

9

nucleotides 

 DBalP Rat 0.25 μmol/mammary gland Bladder 69 Adducts/10

9

nucleotides 

 DBalP Rat 0.25 μmol/mammary gland Liver 116 Adducts/10

9

nucleotides 

 Sum 2,244.63 17630 Cavalieri et 

al., 1981a 

BaP 0.2 μmol/mouse Skin 4 hr 16.3 1 μmol adduct/mol 

DNA 

 CPcdP 0.2 μmol/mouse Skin 4 hr 2.3 0.2 μmol adduct/mol 

DNA 

 ACEP 0.2 μmol/mouse Skin 4 hr 2.2 0.1 μmol adduct/mol 

DNA 
C-84 DRAFT – DO NOT CITE OR QUO

Table C-19. In vivo DNA adducts: dose response data 

DNA adducts 

Record 

number Reference PAH Species Dose Dose units Organ Time Mean 

Standard 

deviation 

Standard 

error Adduct units 

Slope of 

AUC vs. 

dose Comments 

 BaP 0.2 μmol/mouse Skin 24 hr 6.7 1.6 μmol adduct/mol 

DNA 

 CPcdP 0.2 μmol/mouse Skin 24 hr 8.8 1 μmol adduct/mol 

DNA 

 ACEP 0.2 μmol/mouse Skin 24 hr 0.30 0.1 μmol adduct/mol 

DNA 

18810 Hughes 

and Phillips, 1990 

BaP 1 μmol Skin 1 d 7.8 fmol adducts/μg 

DNA 

 Only peak extracted

interrupted scale 

precluded digitizing 

 BaP 1 μmol Lung 2 d 1.2 fmol adducts/μg 

DNA 

 BaP 1 μmol Sum skin 

and lung 

 9.0 fmol adducts/μg 

DNA 

 DBaeP 1 μmol Skin 2 d 0.50 fmol adducts/μg 

DNA 

 DBaeP 1 μmol Lung 7 d Cannot 

determine

 fmol adducts/μg 

DNA 

 DBaeP 1 μmol Sum skin 

and lung 

 Cannot 

determine

 fmol adducts/μg 

DNA 

 DBahP 1 μmol Skin 2 d 3.1 fmol adducts/μg 

DNA 

 DBahP 1 μmol Lung 2 d 0.14 fmol adducts/μg 

DNA 

 DBahP 1 μmol Sum skin 

and lung 

 3.2 fmol adducts/μg 

DNA 

 DBaiP 1 μmol Skin 2 d 0.75 fmol adducts/μg 

DNA 

 DBaiP 1 μmol Lung 2 d 0.10 fmol adducts/μg 

DNA 

 DBaiP 1 μmol Sum skin 

and lung 

 0.85 fmol adducts/μg 

DNA 

 DBalP 1 μmol Skin 1 d 62 fmol adducts/μg 

DNA 

 DBalP 1 μmol Lung 2 d 2.3 fmol adducts/μg 

DNA 
C-85 DRAFT – DO NOT CITE OR QUO

Table C-19. In vivo DNA adducts: dose response data 

DNA adducts 

Record 

number Reference PAH Species Dose Dose units Organ Time Mean 

Standard 

deviation 

Standard 

error Adduct units 

Slope of 

AUC vs. 

dose Comments 

 DBalP 1 μmol Sum skin 

and lung 

 65 fmol adducts/μg 

DNA 

11190 Mass et al., 

1993 

BaP 20 mg/kg bw Lung 24 hr 116 53 amol adducts/μg

DNA 

 AUC calculated usi

trapezoid rule 

 BaP 20 mg/kg bw Lung 48 hr 122 25 amol adducts/μg 

DNA 

 BaP 20 mg/kg bw Lung 72 hr 181 101 amol adducts/μg 

DNA 

 BaP 50 mg/kg bw Lung 24 hr 120 20 amol adducts/μg 

DNA 

 BaP 50 mg/kg bw Lung 48 hr 201 170 amol adducts/μg 

DNA 

 BaP 50 mg/kg bw Lung 72 hr 432 274 amol adducts/μg 

DNA 

 BaP 100 mg/kg bw Lung 24 hr 427 140 amol adducts/μg 

DNA 

 BaP 100 mg/kg bw Lung 48 hr 407 197 amol adducts/μg 

DNA 

 BaP 100 mg/kg bw Lung 72 hr 2,004 314 amol adducts/μg 

DNA 

 BaP 20 mg/kg bw Lung AUC 7,884 469.73  BaP 50 mg/kg bw Lung AUC 12,888  BaP 100 mg/kg bw Lung AUC 44,064  BjAC 20 mg/kg bw Lung 24 hr 63 34 amol adducts/μg 

DNA 

 AUC calculated usi

trapezoid rule 

 BjAC 20 mg/kg bw Lung 48 hr 97 101 amol adducts/μg 

DNA 

 BjAC 20 mg/kg bw Lung 72 hr 255 392 amol adducts/μg 

DNA 

 BjAC 50 mg/kg bw Lung 24 hr 116 121 amol adducts/μg 

DNA 

 BjAC 50 mg/kg bw Lung 48 hr 402 237 amol adducts/μg 

DNA 

 BjAC 50 mg/kg bw Lung 72 hr 1,954 1,921 amol adducts/μg 

DNA 
C-86 DRAFT – DO NOT CITE OR QUO

Table C-19. In vivo DNA adducts: dose response data 

DNA adducts 

Record 

number Reference PAH Species Dose Dose units Organ Time Mean 

Standard 

deviation 

Standard 

error Adduct units 

Slope of 

AUC vs. 

dose Comments 

 BjAC 100 mg/kg bw Lung 24 hr 180 133 amol adducts/μg 

DNA 

 BjAC 100 mg/kg bw Lung 48 hr 532 559 amol adducts/μg 

DNA 

 BjAC 100 mg/kg bw Lung 72 hr 2,439 2,242 amol adducts/μg 

DNA 

 BjAC 20 mg/kg bw Lung AUC 6,900 464.25  BjAC 50 mg/kg bw Lung AUC 35,880  BjAC 100 mg/kg bw Lung AUC 46,356 8010 Nesnow et 

al., 1993b 

BaP 100 mg/kg Lung d 1 453 AUC calculated usi

trapezoid rule 

 BaP 100 mg/kg Lung d 3 1,001  BaP 100 mg/kg Lung d 7 574  BaP 100 mg/kg Lung d 14 386  BaP 100 mg/kg Lung d 28 381  BaP 100 mg/kg Lung d 56 143  BaP 100 mg/kg Lung AUC 20,892  BaP 100 mg/kg Liver d 1 398  BaP 100 mg/kg Liver d 3 1,317  BaP 100 mg/kg Liver d 7 931  BaP 100 mg/kg Liver d 14 537  BaP 100 mg/kg Liver d 28 394  BaP 100 mg/kg Liver d 56 116  BaP 100 mg/kg Liver AUC 25,207  BaP 100 mg/kg PBL d 1 158  BaP 100 mg/kg PBL d 3 273  BaP 100 mg/kg PBL d 7 162  BaP 100 mg/kg PBL d 14 187  BaP 100 mg/kg PBL d 28 72  BaP 100 mg/kg PBL d 56 41  BaP 100 mg/kg PBL AUC 5,985 
C-87 DRAFT – DO NOT CITE OR QUO

Table C-19. In vivo DNA adducts: dose response data 

DNA adducts 

Record 

number Reference PAH Species Dose Dose units Organ Time Mean 

Standard 

deviation 

Standard 

error Adduct units 

Slope of 

AUC vs. 

dose Comments 

 BaP 100 mg/kg Sum of 

AUCs 

 52,084 

 BbF 100 mg/kg Lung d 1 21 AUC calculated usi

trapezoid rule 

 BbF 100 mg/kg Lung d 3 184  BbF 100 mg/kg Lung d 5 233  BbF 100 mg/kg Lung d 7 211  BbF 100 mg/kg Lung d 14 229  BbF 100 mg/kg Lung d 28 145  BbF 100 mg/kg Lung d 56 106  BbF 100 mg/kg Lung AUC 8,763  BbF 100 mg/kg Liver d 1 12  BbF 100 mg/kg Liver d 3 35  BbF 100 mg/kg Liver d 5 51  BbF 100 mg/kg Liver d 7 61  BbF 100 mg/kg Liver d 14 21  BbF 100 mg/kg Liver d 28 15  BbF 100 mg/kg Liver d 56 12  BbF 100 mg/kg Liver AUC 1,173  BbF 100 mg/kg PBL d 1 12  BbF 100 mg/kg PBL d 3 29  BbF 100 mg/kg PBL d 5 59  BbF 100 mg/kg PBL d 7 57  BbF 100 mg/kg PBL d 14 40  BbF 100 mg/kg PBL d 28 15  BbF 100 mg/kg PBL d 56 13  BbF 100 mg/kg PBL AUC 1,378  BbF 100 mg/kg Sum of 

AUCs 

 11,314 

24590/ 20920 

Nesnow et 

al., 1998b; Ross, 1995

BaP NA Lung >21 d 3.9 113 Slope of dose vs. 

TIDAL value (in fmd/μg DNA) 
C-88 DRAFT – DO NOT CITE OR QUO

Table C-19. In vivo DNA adducts: dose response data 

DNA adducts 

Record 

number Reference PAH Species Dose Dose units Organ Time Mean 

Standard 

deviation 

Standard 

error Adduct units 

Slope of 

AUC vs. 

dose Comments 

 BbF NA Lung >21 d 5 37.5 Slope of dose vs. 

TIDAL value (in fmd/μg DNA) 

 CPcdP NA Lung >21 d 3.69 148 Slope of dose vs. 

TIDAL value (in fmd/μg DNA) 

 DBahA NA Lung >21 d 19.1 219 Slope of dose vs. 

TIDAL value (in fmd/μg DNA) 

 DBalP NA Lung >21 d 267 1,390 Slope of dose vs. 

TIDAL value (in fmd/μg DNA) 

22810 Phillips et 

al., 1979 

BaP 1 μmol/mouse Skin 19 hr 27 pmol adducts/mg 

DNA 

 peak 

 DBacA 1 μmol/mouse Skin 24 hr 10 pmol adducts/mg 

DNA 

 peak 

 DBahA 1 μmol/mouse Skin 72 hr 15 pmol adducts/mg 

DNA 

 peak 

24790 Kligerman 

et al., 2002 

BaP Mice 100 mg/kg PBL d 7 4,186 273 amol adducts/μg 

DNA 

 Intraperitoneal 

 BaA Mice 100 mg/kg PBL d 7 93 8 amol adducts/μg 

DNA 

 Intraperitoneal 

 BbF Mice 100 mg/kg PBL d 7 516 7 amol adducts/μg 

DNA 

 Intraperitoneal 

 CH Mice 100 mg/kg PBL d 7 81 11 amol adducts/μg 

DNA 

 Intraperitoneal 

 Control Mice 0 mg/kg PBL d 7 0 amol adducts/μg 

DNA 

 Intraperitoneal 

 BaP Mice 100 mg/kg PBL d 7 143 17 amol adducts/μg 

DNA 

 Gavage 

 BaA Mice 100 mg/kg PBL d 7 32 2 amol adducts/μg 

DNA 

 Gavage 

 BbF Mice 100 mg/kg PBL d 7 39 4 amol adducts/μg 

DNA 

 Gavage 

 CH Mice 100 mg/kg PBL d 7 37 1 amol adducts/μg 

DNA 

 Gavage 
C-89 DRAFT – DO NOT CITE OR QUO

Table C-19. In vivo DNA adducts: dose response data 

DNA adducts 

Record 

number Reference PAH Species Dose Dose units Organ Time Mean 

Standard 

deviation 

Standard 

error Adduct units 

Slope of 

AUC vs. 

dose Comments 

 Control Mice 0 mg/kg PBL d 7 0 amol adducts/μg 

DNA 

 Gavage 

 BaP Rat 100 mg/kg PBL d 7 755 56 amol adducts/μg 

DNA 

 Intraperitoneal 

 BaA Rat 100 mg/kg PBL d 7 38 3 amol adducts/μg 

DNA 

 Intraperitoneal 

 BbF Rat 100 mg/kg PBL d 7 63 1 amol adducts/μg 

DNA 

 Intraperitoneal 

 CH Rat 100 mg/kg PBL d 7 24 2 amol adducts/μg 

DNA 

 Intraperitoneal 

 Control Rat 0 mg/kg PBL d 7 0 amol adducts/μg 

DNA 

 Intraperitoneal 

 BaP Rat 100 mg/kg PBL d 7 177 30 amol adducts/μg 

DNA 

 Gavage 

 BaA Rat 100 mg/kg PBL d 7 20 2 amol adducts/μg 

DNA 

 Gavage 

 BbF Rat 100 mg/kg PBL d 7 17 1 amol adducts/μg 

DNA 

 Gavage 

 CH Rat 100 mg/kg PBL d 7 10 4 amol adducts/μg 

DNA 

 Gavage 

 Control Rat 0 mg/kg PBL d 7 0 amol adducts/μg 

DNA 

 Gavage 
C-90 DRAFT – DO NOT CITE OR QUOTE 

Table C-20. In vivo clastogenicity: data use 

Record 

number Reference Page

Table 

number

Figure 

number PAHs Data to be extracted 

Basis for 

RPF Comment 

24740 Allen et 

al., 1999 

 I and III BaP, 

DBalP

Total micronuleated polychromatic erythrocytes (MNPCE)/PCEs and dose (mg/kg). 

Extract data for bone marrow 

and peripheral blood for both 

A/J mice (Table 1) and p53+/+ 

(wild type) mice (Table III) 

Point 

estimate 

Incidence data. 

Single dose 

BaP. 

14270 He and 

Baker, 

1991 

166 1 BaP, 

CH 

MN cells/1,000 binucleated 

and dose (μg/mouse) 

Ratio of 

slopes 

Incidence data 

17190 Bayer, 

1978 

426 3 BaP, 

PH 

SCE/cells and dose (mg/kg) Point 

estimate 

Continuous 

data. Only one 

dose PH 

significant. 

BaP given as 

3,4-BaP. 

20950 RoszinskyKocher et 

al., 1979 

66 1 BaP, 

DBah

A, 

CH, 

PH, 

BeP, 

BbF, 

BaA 

SCEs/metaphase and dose 

(mg/kg) 

Point 

estimate 

24720 Kligerman 

et al., 1986

129 3 BaP, 

BlAC 

SCEs/metaphase and dose 

(mg/kg) 

Point 

estimate 

Continuous 

data, no SD for 

control; use 

lowest dose 

approaching 

peak 

24790 Kligerman 

et al., 2002

846 1 BaP, 

BaA, 

BbF, 

CH 

SCEs/metaphase, 

intraperitoneal, for BaP, BaA, 

BbF, and CH. SCEs/, gavage, 

for BaP and BaA (use 17.91 

value for BaP). Also use 

MNbn/1,000 bn, gavage, for 

BaP and BbF. Dose in mg/kg. 

Point 

estimates

Separate RPFs 

for SCEs and 

micronuclei, 

oral and 

intraperitoneal 
C-91 DRAFT – DO NOT CITE OR QUO

Table C-21. In vivo clastogenicity: dose response data 

Clastogenicity 

Record 

number Reference PAH 

Route of 

administration Dose Dose units Mean Standard deviation Units n % Response Units 

p < 

0.05 Notes 

24740 Allen et al., 1999 Tricaprylin Intra- peritoneal 0 mg/kg 2.6 MN-PCEs 1,000 0.0026 PCEs A/J mice, bone marrow  BaP Intraperitoneal 200 mg/kg 11.2 MN-PCEs 1,000 0.0112 PCEs x  DBalP Intraperitoneal 0.3 mg/kg 2 MN-PCEs 1,000 0.0020 PCEs  DBalP Intraperitoneal 1.5 mg/kg 3.9 MN-PCEs 1,000 0.0039 PCEs x  DBalP Intraperitoneal 3 mg/kg 3.4 MN-PCEs 1,000 0.0034 PCEs  DBalP Intraperitoneal 6 mg/kg 3.8 MN-PCEs 1,000 0.0038 PCEs  Tricaprylin Intra- peritoneal 0 mg/kg 2.8 MN-PCEs 1,000 0.0028 PCEs A/J mice, peripheral blood  BaP Intraperitoneal 200 mg/kg 9.5 MN-PCEs 1,000 0.0095 PCEs x  DBalP Intraperitoneal 0.3 mg/kg 2.8 MN-PCEs 1,000 0.0028 PCEs  DBalP Intraperitoneal 1.5 mg/kg 2.9 MN-PCEs 1,000 0.0029 PCEs  DBalP Intraperitoneal 3 mg/kg 4 MN-PCEs 1,000 0.0040 PCEs  DBalP Intraperitoneal 6 mg/kg 4.3 MN-PCEs 1,000 0.0043 PCEs x  Tricaprylin Intra- peritoneal 0 mg/kg 3.2 MN-PCEs 1,000 0.0032 PCEs p53 +/+ wt mice, bone marrow  BaP Intraperitoneal 200 mg/kg 5.1 MN-PCEs 1,000 0.0051 PCEs x  DBalP Intraperitoneal 9 mg/kg 4.3 MN-PCEs 1,000 0.0043 PCEs  DBalP Intraperitoneal 12 mg/kg 7.4 MN-PCEs 1,000 0.0074 PCEs x  DBalP Intraperitoneal 18 mg/kg 6.1 MN-PCEs 1,000 0.0061 PCEs x 
C-92 DRAFT – DO NOT CITE OR QUO

Table C-21. In vivo clastogenicity: dose response data 

Clastogenicity 

Record 

number Reference PAH 

Route of 

administration Dose Dose units Mean Standard deviation Units n % Response Units 

p < 

0.05 Notes 

 Tricaprylin Intra- peritoneal 0 mg/kg 3.5 MN-PCEs 1,000 0.0035 PCEs p53 +/+ wt mice, peri

pheral blood 

 BaP Intraperitoneal 200 mg/kg 5.7 MN-PCEs 1,000 0.0057 PCEs x  DBalP Intraperitoneal 9 mg/kg 3.1 MN-PCEs 1,000 0.0031 PCEs  DBalP Intraperitoneal 12 mg/kg 3.1 MN-PCEs 1,000 0.0031 PCEs  DBalP Intraperitoneal 18 mg/kg 4.6 MN-PCEs 1,000 0.0046 PCEs 14270 He and Baker, 1991 Control Dermal 0 μg/mouse 13.3 2.8 MN cells 1,000 0.013 Binucleated BaP Dermal 0.5 μg/mouse 50.5 11.5 MN cells 1,000 0.051 Binucleated x  BaP Dermal 5 μg/mouse 66.8 4.1 MN cells 1,000 0.067 Binucleated x  BaP Dermal 50 μg/mouse 76 2.8 MN cells 1,000 0.076 Binucleated x  BaP Dermal 100 μg/mouse 64.3 5.4 MN cells 1,000 0.064 Binucleated x  BaP Dermal 500 μg/mouse 55.8 13 MN cells 1,000 0.056 Binucleated x  Control Dermal 0 μg/mouse 12.8 2.2 MN cells 1,000 0.013 Binucleated CH Dermal 50 μg/mouse 43.3 2.2 MN cells 1,000 0.043 Binucleated x  CH Dermal 100 μg/mouse 56 4.9 MN cells 1,000 0.056 Binucleated x  CH Dermal 500 μg/mouse 62 8.6 MN cells 1,000 0.062 Binucleated x  CH Dermal 1,000 μg/mouse 47.3 3.8 MN cells 1,000 0.047 Binucleated x 17190 Bayer, 1978 pooled 

controls 

Intraperitoneal 0 mg/kg 3.2 0.07 SCE/cells 

 BaP Intraperitoneal 2.5 mg/kg 3.4 0.8 SCE/cells  BaP Intraperitoneal 25 mg/kg 3.5 0.2 SCE/cells  BaP Intraperitoneal 40 mg/kg 3.9 0.2 SCE/cells x  BaP Intraperitoneal 50 mg/kg 6.4 0.2 SCE/cells x  BaP Intraperitoneal 75 mg/kg 6.4 0.3 SCE/cells x 
C-93 DRAFT – DO NOT CITE OR QUO

Table C-21. In vivo clastogenicity: dose response data 

Clastogenicity 

Record 

number Reference PAH 

Route of 

administration Dose Dose units Mean Standard deviation Units n % Response Units 

p < 

0.05 Notes 

 BaP Intraperitoneal 100 mg/kg 7.4 0.2 SCE/cells x  PH Intraperitoneal 25 mg/kg 3.5 0.2 SCE/cells Only one dose signific PH Intraperitoneal 50 mg/kg 3.4 0.2 SCE/cells  PH Intraperitoneal 75 mg/kg 3.5 0.2 SCE/cells  PH Intraperitoneal 100 mg/kg 4.1 0.2 SCE/cells x 20950 Roszinsky-Kocher et 

al., 1979 

Control Intraperitoneal 0 mg/kg 3.9 0.9 SCEs/metap hase 

 BaP Intraperitoneal 900 mg/kg 10.6 1.6 SCEs/metap hase x  DBahA Intraperitoneal 900 mg/kg 4.9 0.7 SCEs/ x  CH Intraperitoneal 900 mg/kg 5.1 1 SCEs/ x  PH Intraperitoneal 900 mg/kg 5.5 0.7 SCEs/ x  BeP Intraperitoneal 900 mg/kg 5.5 0.7 SCEs/ x  BbF Intraperitoneal 900 mg/kg 5.6 0.5 SCEs/ x  BaA Intraperitoneal 900 mg/kg 6.1 0.4 SCEs/ x 24720 Kligerman et al., 

1986 

Control Gavage 0 mg/kg 11.9 SCEs/metap

hase 

 BaP Gavage 63 mg/kg 19.4 0.0 SCEs/metap

hase 

 BaP Gavage 252 mg/kg 21.5 1.4 SCEs/metap

hase 

 BaP Gavage 504 mg/kg 21.7 1.4 SCEs/metap

hase 
C-94 DRAFT – DO NOT CITE OR QUO

Table C-21. In vivo clastogenicity: dose response data 

Clastogenicity 

Record 

number Reference PAH 

Route of 

administration Dose Dose units Mean Standard deviation Units n % Response Units 

p < 

0.05 Notes 

 Control Gavage 0 mg/kg 11.0 SCEs/metap

hase 

 BlAC Gavage 32 mg/kg 16.5 3.6 SCEs/metap

hase 

 BlAC Gavage 63 mg/kg 20.5 1.6 SCEs/metap

hase 

 BlAC Gavage 126 mg/kg 27.8 2.6 SCEs/metap

hase 

24790 Kligerman et al., 

2002 

Control Intraperitoneal 0 mg/kg 8.79 1.26 SCEs/ 

 BaP Intraperitoneal 100 mg/kg 21.21 2.93 SCEs/ x  BaA Intraperitoneal 100 mg/kg 14.8 3.16 SCEs/ x  BbF Intraperitoneal 100 mg/kg 22.25 1.45 SCEs/ x  CH Intraperitoneal 100 mg/kg 11.96 1.8 SCEs/ x  Control Gavage 0 mg/kg 11.12 1.5 SCEs/  BaP Gavage 100 mg/kg 17.91 1.49 SCEs/ x  BaA Gavage 100 mg/kg 13.38 1.53 SCEs/ x  Control Gavage 0 mg/kg 6.6 0.9 MN bn 1,000 0.007 Binucleated BaP Gavage 100 mg/kg 9.1 1.8 MN bn 1,000 0.009 Binucleated x  BbF Gavage 100 mg/kg 8.3 0.9 MN bn 1,000 0.008 Binucleated x 
D-1 DRAFT – DO NOT CITE OR QUOTE 

1 APPENDIX D. BENCHMARK DOSE MODELING OUTPUTS 

2 

3 D.1. DERMAL BIOASSAYS 

4 Cav 1983 bap dermal.out.txt 

5 ==================================================================== 

6 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

7 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

8 RPS\MODELING\UNSAVED1.(d) 

9 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

10 DOCUMENTS\PAH RPS\MODELING\UNSAVED1.plt 

11 Tue Jul 05 10:10:03 2005 

12 ==================================================================== 

13 

14 BMDS MODEL RUN 

15 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

16 

17 The form of the probability function is: 

18 

19 P[response] = background + (1-background)*[1-EXP( 

20 -beta1*dose^1-beta2*dose^2)] 

21 

22 The parameter betas are restricted to be positive 

23 

24 

25 Dependent variable = COLUMN2 

26 Independent variable = COLUMN1 

27 

28 Total number of observations = 4 

29 Total number of records with missing values = 0 

30 Total number of parameters in model = 3 

31 Total number of specified parameters = 0 

32 Degree of polynomial = 2 

33 

34 

35 Maximum number of iterations = 250 

36 Relative Function Convergence has been set to: 1e-008 

37 Parameter Convergence has been set to: 1e-008 

38 

39 

40 

41 Default Initial Parameter Values 

42 Background = 0.0155298 

43 Beta(1) = 0 

44 Beta(2) = 0.00204447 

45 

46 

47 Asymptotic Correlation Matrix of Parameter Estimates 

48 

49 ( *** The model parameter(s) -Background 

50 have been estimated at a boundary point, or have been 

51 specified by the user, 

52 and do not appear in the correlation matrix ) 

53 

54 Beta(1) Beta(2) 

55 

56 Beta(1) 1 -0.96 

57 

58 Beta(2) -0.96 1 
D-2 DRAFT – DO NOT CITE OR QUOTE 

1 

2 

3 

4 Parameter Estimates 

5 

6 Variable Estimate Std. Err. 

7 Background 0 NA 

8 Beta(1) 0.0126577 0.0460858 

9 Beta(2) 0.00134916 0.00245743 

10 

11 NA - Indicates that this parameter has hit a bound 

12 implied by some inequality constraint and thus

13 has no standard error. 

14 

15 

16 

17 Analysis of Deviance Table 

18 

19 Model Log(likelihood) Deviance Test DF P-value 

20 Full model -35.0798 

21 Fitted model -36.0272 1.89478 2 0.3878 

22 Reduced model -55.062 39.9644 3 <.0001 

23 

24 AIC: 76.0543 

25 

26 

27 Goodness of Fit 

28 

29 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

30 ----------------------------------------------------------------------- 

31 i: 1 

32 0.0000 0.0000 0.000 0 29 0.000 

33 i: 2 

34 2.2000 0.0338 1.014 2 30 1.007 

35 i: 3 

36 6.6000 0.1326 3.714 2 28 -0.532 

37 i: 4 

38 20.0000 0.5474 16.423 17 30 0.078 

39 

40 Chi-square = 1.95 DF = 2 P-value = 0.3772 

41 

42 

43 Benchmark Dose Computation 

44 

45 Specified effect = 0.1 

46 

47 Risk Type = Extra risk 

48 

49 Confidence level = 0.95 

50 

51 BMD = 5.31398 

52 BMDL = 2.86439 
D-3 DRAFT – DO NOT CITE OR QUOTE 

1 CAVALIERI1983BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\SETS\CAVALIERI1983BAP.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\SETS\CAVALIERI1983BAP.plt 

8 Thu Jun 02 11:12:07 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBaP 

23 Independent variable = doseBaP 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.062817 

40 Beta(1) = 0.0817095 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-4 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 0.0898383 0.0186734 

2 Beta(2) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -41.1202 

14 Fitted model -44.3027 6.36496 3 0.09514 

15 Reduced model -76.9419 71.6434 3 <.0001 

16 

17 AIC: 90.6054 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 29 0.000 

26 i: 2 

27 2.2000 0.1793 5.380 2 30 -0.766 

28 i: 3 

29 6.6000 0.4473 12.524 17 28 0.647 

30 i: 4 

31 20.0000 0.8342 25.025 24 30 -0.247 

32 

33 Chi-square = 5.73 DF = 3 P-value = 0.1253 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 0.1 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 1.17278 

45 

46 BMDL = 0.902296 
D-5 DRAFT – DO NOT CITE OR QUOTE 

1 CAVALIERI1983CPcdP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\SETS\CAVALIERI1983BAP.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\SETS\CAVALIERI1983BAP.plt 

8 Thu Jun 02 11:16:02 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceCPcdP 

23 Independent variable = doseCPcdP 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0 

41 Beta(2) = 4.42193e-005 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) Beta(2) 

52 

53 Beta(1) 1 -0.93 

54 

55 Beta(2) -0.93 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-6 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0 NA 

3 Beta(1) 0.000525849 0.00477908 

4 Beta(2) 3.60995e-005 2.68444e-005 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -27.8865 

16 Fitted model -30.0799 4.38685 2 0.1115 

17 Reduced model -64.1091 72.4452 3 <.0001 

18 

19 AIC: 64.1598 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0000 0.000 0 29 0.000 

28 i: 2 

29 22.2000 0.0290 0.842 2 29 1.416 

30 i: 3 

31 66.6000 0.1773 5.141 2 29 -0.743 

32 i: 4 

33 200.0000 0.7876 22.840 24 29 0.239 

34 

35 Chi-square = 4.25 DF = 2 P-value = 0.1194 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.1 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 47.2296 

47 

48 BMDL = 30.0553 

49 
D-7 DRAFT – DO NOT CITE OR QUOTE 

1 HABS1980BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\SETS\HABS1980BAP.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\SETS\HABS1980BAP.plt 

8 Thu May 26 14:32:01 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBaP 

23 Independent variable = doseBaP 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 1 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0 

41 Beta(2) = 0.151094 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(1) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(2) 

52 

53 Beta(2) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-8 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 0 NA 

2 Beta(2) 0.127547 0.0314898 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -40.3373 

14 Fitted model -41.0055 1.33647 2 0.5126 

15 Reduced model -64.1931 47.7118 2 <.0001 

16 

17 AIC: 84.011 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 35 0.000 

26 i: 2 

27 1.7000 0.3083 10.482 8 34 -0.342 

28 i: 3 

29 2.8000 0.6321 22.124 24 35 0.231 

30 

31 Chi-square = 1.28 DF = 2 P-value = 0.5267 

32 

33 

34 Benchmark Dose Computation 

35 

36 Specified effect = 0.1 

37 

38 Risk Type = Extra risk 

39 

40 Confidence level = 0.95 

41 

42 BMD = 0.908873 

43 

44 BMDL = 0.484748 
D-9 DRAFT – DO NOT CITE OR QUOTE 

1 HABS1980BBF.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\HABS1980BBF.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\HABS1980BBF.plt 

8 Wed May 25 13:00:07 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBbF 

23 Independent variable = doseBbF 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0 

41 Beta(2) = 0.00945627 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(1) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(2) 

52 

53 Beta(2) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-10 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 0 NA 

2 Beta(2) 0.00748156 0.00233324 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -47.5575 

14 Fitted model -48.6255 2.13602 3 0.5447 

15 Reduced model -69.4912 43.8674 3 <.0001 

16 

17 AIC: 99.251 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 35 0.000 

26 i: 2 

27 3.4000 0.0829 3.148 2 38 -0.398 

28 i: 3 

29 5.6000 0.2091 7.110 5 34 -0.375 

30 i: 4 

31 9.2000 0.4691 17.358 20 37 0.287 

32 

33 Chi-square = 2.01 DF = 3 P-value = 0.5711 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 0.1 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 3.75269 

45 

46 BMDL = 2.91511 

47 
D-11 DRAFT – DO NOT CITE OR QUOTE 

1 hoff 1966 dermal bap for dbaep.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\UNSAVED1.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\UNSAVED1.plt 

8 Tue Jul 05 10:20:14 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.124609 

40 Beta(1) = 29.9573 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 ( *** The model parameter(s) -Background 

46 have been estimated at a boundary point, or have been 

47 specified by the user, 

48 and do not appear in the correlation matrix ) 

49 

50 Beta(1) 

51 

52 Beta(1) 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0 NA 

60 Beta(1) 34.3074 7.98663 
D-12 DRAFT – DO NOT CITE OR QUOTE 

1 

2 NA - Indicates that this parameter has hit a bound 

3 implied by some inequality constraint and thus

4 has no standard error. 

5 

6 

7 

8 Analysis of Deviance Table 

9 

10 Model Log(likelihood) Deviance Test DF P-value 

11 Full model -12.4245 

12 Fitted model -12.5735 0.297928 2 0.8616 

13 Reduced model -40.3807 55.9124 2 <.0001 

14 

15 AIC: 27.1469 

16 

17 

18 Goodness of Fit 

19 

20 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

21 ----------------------------------------------------------------------- 

22 i: 1 

23 0.0000 0.0000 0.000 0 20 -1.000 

24 i: 2 

25 0.0500 0.8201 16.402 17 20 0.203 

26 i: 3 

27 0.1000 0.9676 19.353 19 20 -0.563 

28 

29 Chi-square = 0.32 DF = 1 P-value = 0.5717 

30 

31 

32 Benchmark Dose Computation 

33 

34 Specified effect = 0.73 

35 

36 Risk Type = Extra risk 

37 

38 Confidence level = 0.95 

39 

40 BMD = 0.0381647 

41 

42 BMDL = 0.026721 
D-13 DRAFT – DO NOT CITE OR QUOTE 

1 HOFFMANWYNDER966DBAIP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\HOFFMANWYNDER966DBAIP.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\HOFFMANWYNDER966DBAIP.plt 

8 Wed May 25 15:24:49 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceDBaiP 

23 Independent variable = doseDBaiP 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.264818 

40 Beta(1) = 18.4583 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 ( *** The model parameter(s) -Background 

46 have been estimated at a boundary point, or have been 

47 specified by the user, 

48 and do not appear in the correlation matrix ) 

49 

50 Beta(1) 

51 

52 Beta(1) 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0 NA 

60 Beta(1) 25.3832 5.83589 
D-14 DRAFT – DO NOT CITE OR QUOTE 

1 

2 NA - Indicates that this parameter has hit a bound 

3 implied by some inequality constraint and thus

4 has no standard error. 

5 

6 

7 

8 Analysis of Deviance Table 

9 

10 Model Log(likelihood) Deviance Test DF P-value 

11 Full model -16.5742 

12 Fitted model -18.019 2.88957 2 0.2358 

13 Reduced model -39.8916 46.6349 2 <.0001 

14 

15 AIC: 38.0379 

16 

17 

18 Goodness of Fit 

19 

20 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

21 ----------------------------------------------------------------------- 

22 i: 1 

23 0.0000 0.0000 0.000 0 20 0.000 

24 i: 2 

25 0.0500 0.7189 13.660 16 19 0.610 

26 i: 3 

27 0.1000 0.9210 17.499 16 19 -1.084 

28 

29 Chi-square = 3.05 DF = 2 P-value = 0.2174 

30 

31 

32 Benchmark Dose Computation 

33 

34 Specified effect = 0.1 

35 

36 Risk Type = Extra risk 

37 

38 Confidence level = 0.95 

39 

40 BMD = 0.00415079 

41 

42 BMDL = 0.00298234 
D-15 DRAFT – DO NOT CITE OR QUOTE 

1 HOFFMANWYNDER1966BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\HOFFMANWYNDER1966BAP.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\HOFFMANWYNDER1966BAP.plt 

8 Wed May 25 15:13:08 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBaP 

23 Independent variable = doseBaP 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.124609 

40 Beta(1) = 29.9573 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 ( *** The model parameter(s) -Background 

46 have been estimated at a boundary point, or have been 

47 specified by the user, 

48 and do not appear in the correlation matrix ) 

49 

50 Beta(1) 

51 

52 Beta(1) 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0 NA 

60 Beta(1) 34.3074 7.98663 
D-16 DRAFT – DO NOT CITE OR QUOTE 

1 

2 NA - Indicates that this parameter has hit a bound 

3 implied by some inequality constraint and thus

4 has no standard error. 

5 

6 

7 

8 Analysis of Deviance Table 

9 

10 Model Log(likelihood) Deviance Test DF P-value 

11 Full model -12.4245 

12 Fitted model -12.5735 0.297928 2 0.8616 

13 Reduced model -40.3807 55.9124 2 <.0001 

14 

15 AIC: 27.1469 

16 

17 

18 Goodness of Fit 

19 

20 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

21 ----------------------------------------------------------------------- 

22 i: 1 

23 0.0000 0.0000 0.000 0 20 -1.000 

24 i: 2 

25 0.0500 0.8201 16.402 17 20 0.203 

26 i: 3 

27 0.1000 0.9676 19.353 19 20 -0.563 

28 

29 Chi-square = 0.32 DF = 1 P-value = 0.5717 

30 

31 

32 Benchmark Dose Computation 

33 

34 Specified effect = 0.1 

35 

36 Risk Type = Extra risk 

37 

38 Confidence level = 0.95 

39 

40 BMD = 0.00307107 

41 

42 BMDL = 0.00215021 
D-17 DRAFT – DO NOT CITE OR QUOTE 

1 HOFFMANWYNDER1966DBAEF.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\SETS\HOFFMANWYNDER1996DBAEP.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\SETS\HOFFMANWYNDER1996DBAEP.plt 

8 Thu Jun 02 11:25:43 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceDBaeF 

23 Independent variable = doseDBaeF 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.22871 

40 Beta(1) = 29.4444 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 ( *** The model parameter(s) -Background 

46 have been estimated at a boundary point, or have been 

47 specified by the user, 

48 and do not appear in the correlation matrix ) 

49 

50 Beta(1) 

51 

52 Beta(1) 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0 NA 

60 Beta(1) 37.3037 9.04943 
D-18 DRAFT – DO NOT CITE OR QUOTE 

1 

2 NA - Indicates that this parameter has hit a bound 

3 implied by some inequality constraint and thus

4 has no standard error. 

5 

6 

7 

8 Analysis of Deviance Table 

9 

10 Model Log(likelihood) Deviance Test DF P-value 

11 Full model -10.3111 

12 Fitted model -10.7582 0.894194 2 0.6395 

13 Reduced model -38.9521 57.2822 2 <.0001 

14 

15 AIC: 23.5163 

16 

17 

18 Goodness of Fit 

19 

20 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

21 ----------------------------------------------------------------------- 

22 i: 1 

23 0.0000 0.0000 0.000 0 20 0.000 

24 i: 2 

25 0.0500 0.8451 16.058 17 19 0.379 

26 i: 3 

27 0.1000 0.9760 18.544 18 19 -1.224 

28 

29 Chi-square = 1.02 DF = 2 P-value = 0.5995 

30 

31 

32 Benchmark Dose Computation 

33 

34 Specified effect = 0.1 

35 

36 Risk Type = Extra risk 

37 

38 Confidence level = 0.95 

39 

40 BMD = 0.0028244 

41 

42 BMDL = 0.00193834 
D-19 DRAFT – DO NOT CITE OR QUOTE 

1 HOFFMANWYNDER1996DBAEP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\BMDS\HOFF_WYND_DBAEP_COMPLETE.(d) 

5 Gnuplot Plotting File: C:\BMDS\HOFF_WYND_DBAEP_COMPLETE.plt 

6 Wed Jul 27 16:17:30 2005 

7 ==================================================================== 

8 

9 BMDS MODEL RUN 

10 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

11 

12 The form of the probability function is: 

13 

14 P[response] = background + (1-background)*[1-EXP( 

15 -beta1*dose^1)] 

16 

17 The parameter betas are restricted to be positive 

18 

19 

20 Dependent variable = COLUMN2 

21 Independent variable = COLUMN1 

22 

23 Total number of observations = 3 

24 Total number of records with missing values = 0 

25 Total number of parameters in model = 2 

26 Total number of specified parameters = 0 

27 Degree of polynomial = 1 

28 

29 

30 Maximum number of iterations = 250 

31 Relative Function Convergence has been set to: 1e-008 

32 Parameter Convergence has been set to: 1e-008 

33 

34 

35 

36 Default Initial Parameter Values 

37 Background = 0.120514 

38 Beta(1) = 7.53772 

39 

40 

41 Asymptotic Correlation Matrix of Parameter Estimates 

42 

43 ( *** The model parameter(s) -Background 

44 have been estimated at a boundary point, or have been 

45 specified by the user, 

46 and do not appear in the correlation matrix ) 

47 

48 Beta(1) 

49 

50 Beta(1) 1 

51 

52 

53 

54 Parameter Estimates 

55 

56 Variable Estimate Std. Err. 

57 Background 0 NA 

58 Beta(1) 11.2084 3.21468 

59 

60 NA - Indicates that this parameter has hit a bound 
D-20 DRAFT – DO NOT CITE OR QUOTE 

1 implied by some inequality constraint and thus

2 has no standard error. 

3 

4 

5 

6 Analysis of Deviance Table 

7 

8 Model Log(likelihood) Deviance Test DF P-value 

9 Full model -32.4818 

10 Fitted model -33.903 2.84251 2 0.2414 

11 Reduced model -44.2604 23.5572 2 <.0001 

12 

13 AIC: 69.8061 

14 

15 

16 Goodness of Fit 

17 

18 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

19 ----------------------------------------------------------------------- 

20 i: 1 

21 0.0000 0.0000 0.000 0 20 0.000 

22 i: 2 

23 0.0500 0.4290 12.871 16 30 0.426 

24 i: 3 

25 0.1000 0.6740 11.458 9 17 -0.658 

26 

27 Chi-square = 2.95 DF = 2 P-value = 0.2288 

28 

29 

30 Benchmark Dose Computation 

31 

32 Specified effect = 0.1 

33 

34 Risk Type = Extra risk 

35 

36 Confidence level = 0.95 

37 

38 BMD = 0.00940018 

39 

40 BMDL = 0.00681373 
D-21 DRAFT – DO NOT CITE OR QUOTE 

1 LAVOIE1982BbF.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\LAVOIE1982.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\LAVOIE1982.plt 

8 Wed May 25 16:18:48 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBbF 

23 Independent variable = doseBbF 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.253748 

40 Beta(1) = 0.0139485 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-22 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 0.0256902 0.00624424 

2 Beta(2) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -37.2311 

14 Fitted model -41.3599 8.25761 3 0.04098 

15 Reduced model -55.2266 35.991 3 <.0001 

16 

17 AIC: 84.7197 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 20 0.000 

26 i: 2 

27 10.0000 0.2266 4.531 9 20 1.275 

28 i: 3 

29 30.0000 0.5373 10.746 12 20 0.252 

30 i: 4 

31 100.0000 0.9234 18.468 16 20 -1.744 

32 

33 Chi-square = 10.32 DF = 3 P-value = 0.0160 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 0.85 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 73.8461 

45 

46 BMDL = 55.1641 
D-23 DRAFT – DO NOT CITE OR QUOTE 

1 LAVOIE1982BjF.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\SETS\LAVOIE1982.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\SETS\LAVOIE1982.plt 

8 Thu May 26 15:18:06 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBjF 

23 Independent variable = doseBjF 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0505665 

40 Beta(1) = 0.00768856 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 ( *** The model parameter(s) -Background 

46 have been estimated at a boundary point, or have been 

47 specified by the user, 

48 and do not appear in the correlation matrix ) 

49 

50 Beta(1) 

51 

52 Beta(1) 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0 NA 

60 Beta(1) 0.00907208 0.00330195 
D-24 DRAFT – DO NOT CITE OR QUOTE 

1 

2 NA - Indicates that this parameter has hit a bound 

3 implied by some inequality constraint and thus

4 has no standard error. 

5 

6 

7 

8 Analysis of Deviance Table 

9 

10 Model Log(likelihood) Deviance Test DF P-value 

11 Full model -25.9801 

12 Fitted model -26.2675 0.574796 2 0.7502 

13 Reduced model -35.7644 19.5688 2 <.0001 

14 

15 AIC: 54.5349 

16 

17 

18 Goodness of Fit 

19 

20 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

21 ----------------------------------------------------------------------- 

22 i: 1 

23 0.0000 0.0000 0.000 0 20 0.000 

24 i: 2 

25 30.0000 0.2383 4.765 6 20 0.340 

26 i: 3 

27 100.0000 0.5964 11.927 11 20 -0.193 

28 

29 Chi-square = 0.60 DF = 2 P-value = 0.7414 

30 

31 

32 Benchmark Dose Computation 

33 

34 Specified effect = 0.85 

35 

36 Risk Type = Extra risk 

37 

38 Confidence level = 0.95 

39 

40 BMD = 209.116 

41 

42 BMDL = 142.347 
D-25 DRAFT – DO NOT CITE OR QUOTE 

1 LAVOIE1982BkF.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\LAVOIE1982.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\LAVOIE1982.plt 

8 Wed May 25 16:21:19 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBkF 

23 Independent variable = doseBkF 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0504814 

40 Beta(1) = 0.00134342 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-26 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 0.00163117 0.000503161 

2 Beta(2) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -26.4637 

14 Fitted model -27.3094 1.69146 3 0.6388 

15 Reduced model -46.0525 39.1775 3 <.0001 

16 

17 AIC: 56.6189 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 20 0.000 

26 i: 2 

27 30.0000 0.0478 0.955 1 20 0.049 

28 i: 3 

29 100.0000 0.1505 3.010 5 20 0.778 

30 i: 4 

31 1000.0000 0.8043 16.086 15 20 -0.345 

32 

33 Chi-square = 1.93 DF = 3 P-value = 0.5881 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 0.85 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 1163.04 

45 

46 BMDL = 802.998 

47 
D-27 DRAFT – DO NOT CITE OR QUOTE 

1 RAVEH1982CPCDP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\RAVEH1982CPCDP.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\RAVEH1982CPCDP.plt 

8 Wed May 25 16:10:56 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBaP 

23 Independent variable = doseBaP 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.086614 

40 Beta(1) = 0.00379482 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 Background Beta(1) Beta(2) 

47 

48 Background 1 -0.51 0.37 

49 

50 Beta(1) -0.51 1 -0.96 

51 

52 Beta(2) 0.37 -0.96 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0.0898028 0.134663 

60 Beta(1) 0.0034393 0.00547992 
D-28 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(2) 1.91358e-006 2.86516e-005 

2 

3 

4 

5 Analysis of Deviance Table 

6 

7 Model Log(likelihood) Deviance Test DF P-value 

8 Full model -57.7672 

9 Fitted model -57.8738 0.213129 1 0.6443 

10 Reduced model -69.2679 23.0015 3 <.0001 

11 

12 AIC: 121.748 

13 

14 

15 Goodness of Fit 

16 

17 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

18 ----------------------------------------------------------------------- 

19 i: 1 

20 0.0000 0.0898 2.604 3 29 0.167 

21 i: 2 

22 10.0000 0.1207 3.622 3 30 -0.195 

23 i: 3 

24 100.0000 0.3669 10.641 11 29 0.053 

25 i: 4 

26 200.0000 0.5762 16.134 16 28 -0.020 

27 

28 Chi-square = 0.21 DF = 1 P-value = 0.6472 

29 

30 

31 Benchmark Dose Computation 

32 

33 Specified effect = 0.1 

34 

35 Risk Type = Extra risk 

36 

37 Confidence level = 0.95 

38 

39 BMD = 30.1292 

40 

41 BMDL = 19.4197 
D-29 DRAFT – DO NOT CITE OR QUOTE 

1 RAVEH_1982BaP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\RAVEH_1982.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\RAVEH_1982.plt 

8 Wed May 25 16:06:59 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBaP 

23 Independent variable = doseBaP 

24 

25 Total number of observations = 6 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 5 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 4 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 6.01899e+017 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 Beta(4) = 0 

44 

45 

46 Asymptotic Correlation Matrix of Parameter Estimates 

47 

48 ( *** The model parameter(s) -Beta(2) -Beta(3) 

49 have been estimated at a boundary point, or have been 

50 specified by the user, 

51 and do not appear in the correlation matrix ) 

52 

53 Background Beta(1) Beta(4) 

54 

55 Background 1 -0.66 0.27 

56 

57 Beta(1) -0.66 1 -0.52 

58 

59 Beta(4) 0.27 -0.52 1 

60 
D-30 DRAFT – DO NOT CITE OR QUOTE 

1 

2 

3 Parameter Estimates 

4 

5 Variable Estimate Std. Err. 

6 Background 0.132052 0.168781 

7 Beta(1) 0.0479561 0.0133452 

8 Beta(2) 0 NA 

9 Beta(3) 0 NA 

10 Beta(4) 4.58924e-009 3.25141e-008 

11 

12 NA - Indicates that this parameter has hit a bound 

13 implied by some inequality constraint and thus

14 has no standard error. 

15 

16 

17 

18 Analysis of Deviance Table 

19 

20 Model Log(likelihood) Deviance Test DF P-value 

21 Full model -56.5419 

22 Fitted model -58.376 3.66814 3 0.2996 

23 Reduced model -101.065 89.0461 5 <.0001 

24 

25 AIC: 122.752 

26 

27 

28 Goodness of Fit 

29 

30 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

31 ----------------------------------------------------------------------- 

32 i: 1 

33 0.0000 0.1321 3.830 3 29 -0.250 

34 i: 2 

35 10.0000 0.4627 13.419 17 29 0.497 

36 i: 3 

37 25.0000 0.7388 20.685 21 28 0.058 

38 i: 4 

39 50.0000 0.9233 25.853 24 28 -0.935 

40 i: 5 

41 100.0000 0.9955 26.878 27 27 1.005 

42 i: 6 

43 200.0000 1.0000 26.000 26 26 1.000 

44 

45 Chi-square = 3.86 DF = 3 P-value = 0.2771 

46 

47 

48 Benchmark Dose Computation 

49 

50 Specified effect = 0.1 

51 

52 Risk Type = Extra risk 

53 

54 Confidence level = 0.95 

55 

56 BMD = 2.19702 

57 

58 BMDL = 1.66278 

59 
D-31 DRAFT – DO NOT CITE OR QUOTE 

1 RICE1988INITIATIONBbcAC.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

5 STUDIES\RICE1988INITIATION.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\DERMAL 

7 STUDIES\RICE1988INITIATION.plt 

8 Thu May 26 12:50:09 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBbcAC 

23 Independent variable = doseBbcAC 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.515248 

40 Beta(1) = 0.484044 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Background Beta(1) 

52 

53 Background 1 -0.68 

54 

55 Beta(1) -0.68 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-32 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0.110219 0.243438 

3 Beta(1) 1.01652 0.319132 

4 Beta(2) 0 NA 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -28.2203 

16 Fitted model -34.2145 11.9884 2 0.002493 

17 Reduced model -51.7957 47.1508 3 <.0001 

18 

19 AIC: 72.4291 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.1102 2.204 1 20 -0.614 

28 i: 2 

29 0.5000 0.4648 9.295 15 20 1.147 

30 i: 3 

31 2.0000 0.8835 17.670 18 20 0.160 

32 i: 4 

33 4.0000 0.9847 19.695 18 20 -5.641 

34 

35 Chi-square = 16.90 DF = 2 P-value = 0.0002 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.89 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 2.1714 

47 

48 BMDL = 1.54343 
D-33 DRAFT – DO NOT CITE OR QUOTE 

1 Rice 1988CH.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\BMDS\UNSAVED1.(d) 

5 Gnuplot Plotting File: C:\BMDS\UNSAVED1.plt 

6 Thu May 26 12:43:53 2005 

7 ==================================================================== 

8 

9 BMDS MODEL RUN 

10 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

11 

12 The form of the probability function is: 

13 

14 P[response] = background + (1-background)*[1-EXP( 

15 -beta1*dose^1-beta2*dose^2)] 

16 

17 The parameter betas are restricted to be positive 

18 

19 

20 Dependent variable = incidenceCH 

21 Independent variable = doseCH 

22 

23 Total number of observations = 4 

24 Total number of records with missing values = 0 

25 Total number of parameters in model = 3 

26 Total number of specified parameters = 0 

27 Degree of polynomial = 2 

28 

29 

30 Maximum number of iterations = 250 

31 Relative Function Convergence has been set to: 1e-008 

32 Parameter Convergence has been set to: 1e-008 

33 

34 

35 

36 Default Initial Parameter Values 

37 Background = 0.305186 

38 Beta(1) = 1.94458 

39 Beta(2) = 0 

40 

41 

42 Asymptotic Correlation Matrix of Parameter Estimates 

43 

44 ( *** The model parameter(s) -Beta(2) 

45 have been estimated at a boundary point, or have been 

46 specified by the user, 

47 and do not appear in the correlation matrix ) 

48 

49 Background Beta(1) 

50 

51 Background 1 -0.51 

52 

53 Beta(1) -0.51 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0.0435025 0.182669 
D-34 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 2.7068 0.74964 

2 Beta(2) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -25.689 

14 Fitted model -28.1343 4.89062 2 0.0867 

15 Reduced model -55.2266 59.0752 3 <.0001 

16 

17 AIC: 60.2686 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0435 0.870 1 20 0.156 

26 i: 2 

27 0.1500 0.3627 7.254 5 20 -0.488 

28 i: 3 

29 0.5000 0.7529 15.058 18 20 0.791 

30 i: 4 

31 1.5000 0.9835 19.670 19 20 -2.065 

32 

33 Chi-square = 4.83 DF = 2 P-value = 0.0894 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 0.89 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 0.815455 

45 

46 BMDL = 0.584044 
D-35 DRAFT – DO NOT CITE OR QUOTE 

1 RICE_CPDEFC.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\RICE_DERMAL_CPDEFC.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\RICE_DERMAL_CPDEFC.plt 

8 Thu Jun 30 20:30:27 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 1 

40 Beta(1) = 6.76726e+019 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Beta(1) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Background Beta(2) 

52 

53 Background 1 -0.52 

54 

55 Beta(2) -0.52 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-36 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0.0499932 0.217763 

3 Beta(1) 0 NA 

4 Beta(2) 44.3918 19.5918 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -16.9192 

16 Fitted model -16.9195 0.000547543 2 0.9997 

17 Reduced model -49.6481 65.4577 3 <.0001 

18 

19 AIC: 37.839 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0500 1.000 1 20 0.000 

28 i: 2 

29 0.1500 0.6501 13.002 13 20 -0.000 

30 i: 3 

31 0.5000 1.0000 19.000 19 19 1.000 

32 i: 4 

33 1.5000 1.0000 19.000 19 19 0.000 

34 

35 Chi-square = 0.00 DF = 2 P-value = 0.9999 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.88 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 0.218546 

47 

48 BMDL = 0.172781 

49 
D-37 DRAFT – DO NOT CITE OR QUOTE 

1 NESNOW_1984_DERMAL_BLAC_MALE.txt

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\_PAH 

5 RPS\MODELING\NESNOW_1984_DERMAL_BLAC_MALE.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\_PAH RPS\MODELING\NESNOW_1984_DERMAL_BLAC_MALE.plt 

8 Thu Feb 08 09:10:48 2007 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0.0283321 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 ( *** The model parameter(s) -Background 

46 have been estimated at a boundary point, or have been 

47 specified by the user, 

48 and do not appear in the correlation matrix ) 

49 

50 Beta(1) 

51 

52 Beta(1) 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0 NA 

60 Beta(1) 0.0219722 0.00534523 
D-38 DRAFT – DO NOT CITE OR QUOTE 

1 

2 NA - Indicates that this parameter has hit a bound 

3 implied by some inequality constraint and thus

4 has no standard error. 

5 

6 

7 

8 Analysis of Deviance Table 

9 

10 Model Log(likelihood) Deviance Test DF P-value 

11 Full model -17.2634 

12 Fitted model -17.7362 0.945584 2 0.6233 

13 Reduced model -39.5006 44.4744 2 <.0001 

14 

15 AIC: 37.4725 

16 

17 

18 Goodness of Fit 

19 

20 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

21 ----------------------------------------------------------------------- 

22 i: 1 

23 0.0000 0.0000 0.000 0 20 0.000 

24 i: 2 

25 50.0000 0.6667 13.333 12 20 -0.300 

26 i: 3 

27 100.0000 0.8889 15.111 16 17 0.529 

28 

29 Chi-square = 0.87 DF = 2 P-value = 0.6471 

30 

31 

32 Benchmark Dose Computation 

33 

34 Specified effect = 0.67 

35 

36 Risk Type = Extra risk 

37 

38 Confidence level = 0.95 

39 

40 BMD = 50.4574 

41 

42 BMDL = 35.8134 

43 
D-39 DRAFT – DO NOT CITE OR QUOTE 

1 NESNOW_1984_DERMAL_BLAC_FEMALE.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\_PAH 

5 RPS\MODELING\NESNOW_1984_DERMAL_BLAC_FEMALE.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\_PAH RPS\MODELING\NESNOW_1984_DERMAL_BLAC_FEMALE.plt 

8 Thu Feb 08 09:13:51 2007 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0.0289037 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 Background Beta(1) 

46 

47 Background 1 -0.49 

48 

49 Beta(1) -0.49 1 

50 

51 

52 

53 Parameter Estimates 

54 

55 Variable Estimate Std. Err. 

56 Background 0.05051 0.21268 

57 Beta(1) 0.0234714 0.00648098 

58 

59 

60 
D-40 DRAFT – DO NOT CITE OR QUOTE 

1 Analysis of Deviance Table 

2 

3 Model Log(likelihood) Deviance Test DF P-value 

4 Full model -20.7842 

5 Fitted model -21.1281 0.687832 1 0.4069 

6 Reduced model -39.8916 38.2148 2 <.0001 

7 

8 AIC: 46.2563 

9 

10 

11 Goodness of Fit 

12 

13 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

14 ----------------------------------------------------------------------- 

15 i: 1 

16 0.0000 0.0505 0.960 1 19 0.044 

17 i: 2 

18 50.0000 0.7064 14.127 13 20 -0.272 

19 i: 3 

20 100.0000 0.9092 17.275 18 19 0.462 

21 

22 Chi-square = 0.64 DF = 1 P-value = 0.4224 

23 

24 

25 Benchmark Dose Computation 

26 

27 Specified effect = 0.51 

28 

29 Risk Type = Extra risk 

30 

31 Confidence level = 0.95 

32 

33 BMD = 30.3924 

34 

35 BMDL = 21.4681 
D-41 DRAFT – DO NOT CITE OR QUOTE 

1 NESNOW_1984_DERMAL_BEAC_MALE.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\_PAH 

5 RPS\MODELING\NESNOW_1984_DERMAL_BEAC_MALE.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\_PAH RPS\MODELING\NESNOW_1984_DERMAL_BEAC_MALE.plt 

8 Fri Feb 09 10:09:40 2007 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 6 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 5 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 4 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.121669 

40 Beta(1) = 0.00219353 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 Beta(4) = 0 

44 

45 

46 Asymptotic Correlation Matrix of Parameter Estimates 

47 

48 ( *** The model parameter(s) -Background -Beta(2) -Beta(3) 

49 -Beta(4) 

50 have been estimated at a boundary point, or have been 

51 specified by the user, 

52 and do not appear in the correlation matrix ) 

53 

54 Beta(1) 

55 

56 Beta(1) 1 

57 

58 

59 

60 Parameter Estimates 
D-42 DRAFT – DO NOT CITE OR QUOTE 

1 

2 Variable Estimate Std. Err. 

3 Background 0 NA 

4 Beta(1) 0.00282301 0.00056435 

5 Beta(2) 0 NA 

6 Beta(3) 0 NA 

7 Beta(4) 0 NA 

8 

9 NA - Indicates that this parameter has hit a bound 

10 implied by some inequality constraint and thus

11 has no standard error. 

12 

13 

14 

15 Analysis of Deviance Table 

16 

17 Model Log(likelihood) Deviance Test DF P-value 

18 Full model -51.002 

19 Fitted model -52.1858 2.36767 5 0.7963 

20 Reduced model -80.7033 59.4025 5 <.0001 

21 

22 AIC: 106.372 

23 

24 

25 Goodness of Fit 

26 

27 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

28 ----------------------------------------------------------------------- 

29 i: 1 

30 0.0000 0.0000 0.000 0 20 0.000 

31 i: 2 

32 50.0000 0.1316 2.633 4 20 0.598 

33 i: 3 

34 100.0000 0.2460 4.919 4 20 -0.248 

35 i: 4 

36 250.0000 0.5063 10.125 12 20 0.375 

37 i: 5 

38 500.0000 0.7562 15.124 15 20 -0.034 

39 i: 6 

40 1000.0000 0.9406 16.930 16 18 -0.925 

41 

42 Chi-square = 2.61 DF = 5 P-value = 0.7594 

43 

44 

45 Benchmark Dose Computation 

46 

47 Specified effect = 0.67 

48 

49 Risk Type = Extra risk 

50 

51 Confidence level = 0.95 

52 

53 BMD = 392.724 

54 

55 BMDL = 305.587 
D-43 DRAFT – DO NOT CITE OR QUOTE 

1 NESNOW_1984_DERMAL_BEAC_FEMALE.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\_PAH 

5 RPS\MODELING\NESNOW_1984_DERMAL_BEAC_FEMALE.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\_PAH RPS\MODELING\NESNOW_1984_DERMAL_BEAC_FEMALE.plt 

8 Fri Feb 09 10:12:31 2007 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0667811 

40 Beta(1) = 0.00288357 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-44 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 0.00366005 0.00126417 

2 Beta(2) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -35.6556 

14 Fitted model -36.1032 0.895174 3 0.8266 

15 Reduced model -45.1184 18.9255 3 0.0002833 

16 

17 AIC: 74.2064 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 19 0.000 

26 i: 2 

27 50.0000 0.1672 3.345 4 20 0.235 

28 i: 3 

29 100.0000 0.3065 5.824 7 19 0.291 

30 i: 4 

31 250.0000 0.5995 11.390 10 19 -0.305 

32 

33 Chi-square = 0.92 DF = 3 P-value = 0.8205 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 0.51 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 194.902 

45 

46 BMDL = 137.872 
D-45 DRAFT – DO NOT CITE OR QUOTE 

1 D.2. INTRAPERITONEAL BIOASSAYS 

2 lavoie 1994 female lung FA.txt 

3 ==================================================================== 

4 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

5 Input Data File: C:\BMDS\UNSAVED1.(d) 

6 Gnuplot Plotting File: C:\BMDS\UNSAVED1.plt 

7 Thu Jun 30 15:55:31 2005 

8 ==================================================================== 

9 

10 BMDS MODEL RUN 

11 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

12 

13 The form of the probability function is: 

14 

15 P[response] = background + (1-background)*[1-EXP( 

16 -beta1*dose^1)] 

17 

18 The parameter betas are restricted to be positive 

19 

20 

21 Dependent variable = COLUMN2 

22 Independent variable = COLUMN1 

23 

24 Total number of observations = 3 

25 Total number of records with missing values = 0 

26 Total number of parameters in model = 2 

27 Total number of specified parameters = 0 

28 Degree of polynomial = 1 

29 

30 

31 Maximum number of iterations = 250 

32 Relative Function Convergence has been set to: 1e-008 

33 Parameter Convergence has been set to: 1e-008 

34 

35 

36 

37 Default Initial Parameter Values 

38 Background = 0.0929049 

39 Beta(1) = 0.108473 

40 

41 

42 Asymptotic Correlation Matrix of Parameter Estimates 

43 

44 Background Beta(1) 

45 

46 Background 1 -0.48 

47 

48 Beta(1) -0.48 1 

49 

50 

51 

52 Parameter Estimates 

53 

54 Variable Estimate Std. Err. 

55 Background 0.112497 0.137421 

56 Beta(1) 0.103015 0.0291539 

57 

58 

59 
D-46 DRAFT – DO NOT CITE OR QUOTE 

1 Analysis of Deviance Table 

2 

3 Model Log(likelihood) Deviance Test DF P-value 

4 Full model -44.1118 

5 Fitted model -44.1689 0.114322 1 0.7353 

6 Reduced model -64.1094 39.9952 2 <.0001 

7 

8 AIC: 92.3379 

9 

10 

11 Goodness of Fit 

12 

13 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

14 ----------------------------------------------------------------------- 

15 i: 1 

16 0.0000 0.1125 3.825 4 34 0.052 

17 i: 2 

18 3.4600 0.3786 11.737 11 31 -0.101 

19 i: 3 

20 17.3000 0.8507 24.669 25 29 0.090 

21 

22 Chi-square = 0.11 DF = 1 P-value = 0.7366 

23 

24 

25 Benchmark Dose Computation 

26 

27 Specified effect = 0.83 

28 

29 Risk Type = Extra risk 

30 

31 Confidence level = 0.95 

32 

33 BMD = 17.201 

34 

35 BMDL = 12.2186 
D-47 DRAFT – DO NOT CITE OR QUOTE 

1 LAVOIEETAL1994LIVERmale.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD 

5 ANALYSIS\BIOASSAY\INTRAPERITONEAL\SETS\LAVOIEETAL1994LIVER.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD 

7 ANALYSIS\BIOASSAY\INTRAPERITONEAL\SETS\LAVOIEETAL1994LIVER.plt 

8 Wed Jun 01 09:15:04 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = response 

23 Independent variable = dose 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 6.19323e+018 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 Background Beta(1) 

46 

47 Background 1 -0.47 

48 

49 Beta(1) -0.47 1 

50 

51 

52 

53 Parameter Estimates 

54 

55 Variable Estimate Std. Err. 

56 Background 0.168707 0.160946 

57 Beta(1) 0.259821 0.0902001 

58 

59 

60 
D-48 DRAFT – DO NOT CITE OR QUOTE 

1 Analysis of Deviance Table 

2 

3 Model Log(likelihood) Deviance Test DF P-value 

4 Full model -31.5803 

5 Fitted model -31.7622 0.363803 1 0.5464 

6 Reduced model -51.0494 38.9382 2 <.0001 

7 

8 AIC: 67.5244 

9 

10 

11 Goodness of Fit 

12 

13 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

14 ----------------------------------------------------------------------- 

15 i: 1 

16 0.0000 0.1687 4.892 5 29 0.026 

17 i: 2 

18 3.4600 0.6617 18.527 18 28 -0.084 

19 i: 3 

20 17.3000 0.9907 16.842 17 17 1.009 

21 

22 Chi-square = 0.21 DF = 1 P-value = 0.6496 

23 

24 

25 Benchmark Dose Computation 

26 

27 Specified effect = 0.81 

28 

29 Risk Type = Extra risk 

30 

31 Confidence level = 0.95 

32 

33 BMD = 6.39183 

34 

35 BMDL = 4.18834 
D-49 DRAFT – DO NOT CITE OR QUOTE 

1 LAVOIEETAL1994LUNGmale.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD 

5 ANALYSIS\BIOASSAY\INTRAPERITONEAL\SETS\LAVOIEETAL1994LUNG.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD 

7 ANALYSIS\BIOASSAY\INTRAPERITONEAL\SETS\LAVOIEETAL1994LUNG.plt 

8 Wed Jun 01 09:49:11 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = responsemale 

23 Independent variable = dosemale 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.24757 

40 Beta(1) = 0.0451334 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 Background Beta(1) 

46 

47 Background 1 -0.57 

48 

49 Beta(1) -0.57 1 

50 

51 

52 

53 Parameter Estimates 

54 

55 Variable Estimate Std. Err. 

56 Background 0.209483 0.142769 

57 Beta(1) 0.0559823 0.0297979 

58 

59 

60 
D-50 DRAFT – DO NOT CITE OR QUOTE 

1 Analysis of Deviance Table 

2 

3 Model Log(likelihood) Deviance Test DF P-value 

4 Full model -43.4897 

5 Fitted model -44.1071 1.23476 1 0.2665 

6 Reduced model -49.0816 11.1837 2 0.003728 

7 

8 AIC: 92.2143 

9 

10 

11 Goodness of Fit 

12 

13 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

14 ----------------------------------------------------------------------- 

15 i: 1 

16 0.0000 0.2095 6.075 5 29 -0.224 

17 i: 2 

18 3.4600 0.3487 9.763 12 28 0.352 

19 i: 3 

20 17.3000 0.6999 11.898 11 17 -0.251 

21 

22 Chi-square = 1.25 DF = 1 P-value = 0.2629 

23 

24 

25 Benchmark Dose Computation 

26 

27 Specified effect = 0.7 

28 

29 Risk Type = Extra risk 

30 

31 Confidence level = 0.95 

32 

33 BMD = 21.5063 

34 

35 BMDL = 12.5156 
D-51 DRAFT – DO NOT CITE OR QUOTE 

1 WISLOCKI_CHRYSENE_MALE_LIVER.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\BMDS\WISLOCKI_CHRYSENE_MALE_LIVER.(d) 

5 Gnuplot Plotting File: C:\BMDS\WISLOCKI_CHRYSENE_MALE_LIVER.plt 

6 Wed Jun 15 13:20:42 2005 

7 ==================================================================== 

8 

9 BMDS MODEL RUN 

10 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

11 

12 The form of the probability function is: 

13 

14 P[response] = background + (1-background)*[1-EXP( 

15 -beta1*dose^1)] 

16 

17 The parameter betas are restricted to be positive 

18 

19 

20 Dependent variable = COLUMN2 

21 Independent variable = COLUMN1 

22 

23 Total number of observations = 3 

24 Total number of records with missing values = 0 

25 Total number of parameters in model = 2 

26 Total number of specified parameters = 0 

27 Degree of polynomial = 1 

28 

29 

30 Maximum number of iterations = 250 

31 Relative Function Convergence has been set to: 1e-008 

32 Parameter Convergence has been set to: 1e-008 

33 

34 

35 

36 Default Initial Parameter Values 

37 Background = 0.147839 

38 Beta(1) = 0.000139419 

39 

40 

41 Asymptotic Correlation Matrix of Parameter Estimates 

42 

43 Background Beta(1) 

44 

45 Background 1 -0.57 

46 

47 Beta(1) -0.57 1 

48 

49 

50 

51 Parameter Estimates 

52 

53 Variable Estimate Std. Err. 

54 Background 0.109703 0.10278 

55 Beta(1) 0.000173669 9.88799e-005 

56 

57 

58 

59 Analysis of Deviance Table 

60 
D-52 DRAFT – DO NOT CITE OR QUOTE 

1 Model Log(likelihood) Deviance Test DF P-value 

2 Full model -67.0392 

3 Fitted model -67.7628 1.44719 1 0.229 

4 Reduced model -74.516 14.9536 2 0.0005661 

5 

6 AIC: 139.526 

7 

8 

9 Goodness of Fit 

10 

11 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

12 ----------------------------------------------------------------------- 

13 i: 1 

14 0.0000 0.1097 8.008 7 73 -0.141 

15 i: 2 

16 700.0000 0.2116 7.407 10 35 0.444 

17 i: 3 

18 2800.0000 0.4525 15.387 14 34 -0.165 

19 

20 Chi-square = 1.52 DF = 1 P-value = 0.2172 

21 

22 

23 Benchmark Dose Computation 

24 

25 Specified effect = 0.44 

26 

27 Risk Type = Extra risk 

28 

29 Confidence level = 0.95 

30 

31 BMD = 3338.63 

32 

33 BMDL = 2098.51 
D-53 DRAFT – DO NOT CITE OR QUOTE 

1 WISLOCKI_CHRYSENE_MALE_LUNG.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\BMDS\WISLOCKI_CHRYSENE_MALE_LUNG.(d) 

5 Gnuplot Plotting File: C:\BMDS\WISLOCKI_CHRYSENE_MALE_LUNG.plt 

6 Wed Jun 15 13:21:42 2005 

7 ==================================================================== 

8 

9 BMDS MODEL RUN 

10 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

11 

12 The form of the probability function is: 

13 

14 P[response] = background + (1-background)*[1-EXP( 

15 -beta1*dose^1)] 

16 

17 The parameter betas are restricted to be positive 

18 

19 

20 Dependent variable = COLUMN2 

21 Independent variable = COLUMN1 

22 

23 Total number of observations = 3 

24 Total number of records with missing values = 0 

25 Total number of parameters in model = 2 

26 Total number of specified parameters = 0 

27 Degree of polynomial = 1 

28 

29 

30 Maximum number of iterations = 250 

31 Relative Function Convergence has been set to: 1e-008 

32 Parameter Convergence has been set to: 1e-008 

33 

34 

35 

36 Default Initial Parameter Values 

37 Background = 0.101102 

38 Beta(1) = 4.85056e-005 

39 

40 

41 Asymptotic Correlation Matrix of Parameter Estimates 

42 

43 Background Beta(1) 

44 

45 Background 1 -0.6 

46 

47 Beta(1) -0.6 1 

48 

49 

50 

51 Parameter Estimates 

52 

53 Variable Estimate Std. Err. 

54 Background 0.0806675 0.103469 

55 Beta(1) 6.36834e-005 8.767e-005 

56 

57 

58 

59 Analysis of Deviance Table 

60 
D-54 DRAFT – DO NOT CITE OR QUOTE 

1 Model Log(likelihood) Deviance Test DF P-value 

2 Full model -51.5522 

3 Fitted model -52.0709 1.03747 1 0.3084 

4 Reduced model -53.9858 4.86735 2 0.08771 

5 

6 AIC: 108.142 

7 

8 

9 Goodness of Fit 

10 

11 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

12 ----------------------------------------------------------------------- 

13 i: 1 

14 0.0000 0.0807 5.889 5 73 -0.164 

15 i: 2 

16 700.0000 0.1207 4.226 6 35 0.477 

17 i: 3 

18 2800.0000 0.2308 7.848 7 34 -0.140 

19 

20 Chi-square = 1.11 DF = 1 P-value = 0.2917 

21 

22 

23 Benchmark Dose Computation 

24 

25 Specified effect = 0.3 

26 

27 Risk Type = Extra risk 

28 

29 Confidence level = 0.95 

30 

31 BMD = 5600.76 

32 

33 BMDL = 2691.64 
D-55 DRAFT – DO NOT CITE OR QUOTE 

1 Busby 1984 i.p. multiplicity 

2 FA male 

3 Linear 

4 Nonconstant variance 

5 BMR = lowest statistically significant response in BaP treated animals (after 

6 control subtracted) 

7 

8 ==================================================================== 

9 Polynomial Model. (Version: 2.12; Date: 02/20/2007) 

10 Input Data File: C:\BMDS\UNSAVED1.(d) 

11 Gnuplot Plotting File: C:\BMDS\UNSAVED1.plt 

12 Mon May 11 21:08:40 2009 

13 ==================================================================== 

14 

15 BMDS MODEL RUN 

16 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

17 

18 The form of the response function is: 

19 

20 Y[dose] = beta_0 + beta_1*dose + beta_2*dose^2 + ... 

21 

22 

23 Dependent variable = MEAN 

24 Independent variable = COLUMN1 

25 The polynomial coefficients are restricted to be positive 

26 The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho) 

27 

28 Total number of dose groups = 3 

29 Total number of records with missing values = 0 

30 Maximum number of iterations = 250 

31 Relative Function Convergence has been set to: 1e-008 

32 Parameter Convergence has been set to: 1e-008 

33 

34 

35 

36 Default Initial Parameter Values 

37 lalpha = 0.136152 

38 rho = 0 

39 beta_0 = 0.0180952 

40 beta_1 = 0.427551 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 lalpha rho beta_0 beta_1 

46 

47 lalpha 1 0.65 0.015 0.00041 

48 

49 rho 0.65 1 0.22 -0.061 

50 

51 beta_0 0.015 0.22 1 -0.24 

52 

53 beta_1 0.00041 -0.061 -0.24 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 95.0% Wald 

60 Confidence Interval 
D-56 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. Lower Conf. Limit 

2 Upper Conf. Limit 

3 lalpha 0.634298 0.204652 0.233188 

4 1.03541 

5 rho 0.923372 0.0876305 0.751619 

6 1.09512 

7 beta_0 0.0170376 0.0434041 -0.0680328 

8 0.102108 

9 beta_1 0.426604 0.0861283 0.257796 

10 0.595413 

11 

12 

13 

14 Table of Data and Estimated Values of Interest

15 

16 Dose N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled 

17 Res. 

18 ------ --- -------- -------- ----------- ----------- ---------

19 - 

20 

21 0 27 0.04 0.017 0.21 0.21 0.57 

22 0.7 31 0.29 0.316 0.84 0.806 -0.177 

23 3.5 27 1.52 1.51 1.66 1.66 0.0308 

24 

25 

26 

27 Model Descriptions for likelihoods calculated 

28 

29 

30 Model A1: Yij = Mu(i) + e(ij) 

31 Var{e(ij)} = Sigma^2 

32 

33 Model A2: Yij = Mu(i) + e(ij) 

34 Var{e(ij)} = Sigma(i)^2 

35 

36 Model A3: Yij = Mu(i) + e(ij) 

37 Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))

38 Model A3 uses any fixed variance parameters that 

39 were specified by the user 

40 

41 Model R: Yi = Mu + e(i) 

42 Var{e(i)} = Sigma^2 

43 

44 

45 Likelihoods of Interest 

46 

47 Model Log(likelihood) # Param's AIC 

48 A1 -46.759351 4 101.518703 

49 A2 -7.114400 6 26.228800 

50 A3 -7.317284 5 24.634569 

51 fitted -7.329046 4 22.658093 

52 R -59.984569 2 123.969139 

53 

54 

55 Explanation of Tests 

56 

57 Test 1: Do responses and/or variances differ among Dose levels? 

58 (A2 vs. R) 

59 Test 2: Are Variances Homogeneous? (A1 vs A2) 

60 Test 3: Are variances adequately modeled? (A2 vs. A3) 
D-57 DRAFT – DO NOT CITE OR QUOTE 

1 Test 4: Does the Model for the Mean Fit? (A3 vs. fitted) 

2 (Note: When rho=0 the results of Test 3 and Test 2 will be the same.) 

3 

4 Tests of Interest 

5 

6 Test -2*log(Likelihood Ratio) Test df p-value 

7 

8 Test 1 105.74 4 <.0001 

9 Test 2 79.2899 2 <.0001 

10 Test 3 0.405769 1 0.5241 

11 Test 4 0.0235238 1 0.8781 

12 

13 The p-value for Test 1 is less than .05. There appears to be a 

14 difference between response and/or variances among the dose levels 

15 It seems appropriate to model the data 

16 

17 The p-value for Test 2 is less than .1. A non-homogeneous variance 

18 model appears to be appropriate 

19 

20 The p-value for Test 3 is greater than .1. The modeled variance appears 

21 to be appropriate here 

22 

23 The p-value for Test 4 is greater than .1. The model chosen seems 

24 to adequately describe the data 

25 

26 

27 Benchmark Dose Computation 

28 

29 Specified effect = 4.28 

30 

31 Risk Type = Point risk 

32 

33 Confidence level = 0.95 

34 

35 BMD = 9.99278 

36 

37 

38 BMDL = 7.55762 
D-58 DRAFT – DO NOT CITE OR QUOTE 

1 Busby 1984 i.p. multiplicity 

2 FA female 

3 Linear 

4 Nonconstant variance 

5 BMR = lowest statistically significant response in BaP treated animals (after 

6 control subtracted) 

7 

8 ==================================================================== 

9 Polynomial Model. (Version: 2.12; Date: 02/20/2007) 

10 Input Data File: C:\BMDS\UNSAVED1.(d) 

11 Gnuplot Plotting File: C:\BMDS\UNSAVED1.plt 

12 Mon May 11 21:14:08 2009 

13 ==================================================================== 

14 

15 BMDS MODEL RUN 

16 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

17 

18 The form of the response function is: 

19 

20 Y[dose] = beta_0 + beta_1*dose + beta_2*dose^2 + ... 

21 

22 

23 Dependent variable = MEAN 

24 Independent variable = COLUMN1 

25 The polynomial coefficients are restricted to be positive 

26 The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho) 

27 

28 Total number of dose groups = 3 

29 Total number of records with missing values = 0 

30 Maximum number of iterations = 250 

31 Relative Function Convergence has been set to: 1e-008 

32 Parameter Convergence has been set to: 1e-008 

33 

34 

35 

36 Default Initial Parameter Values 

37 lalpha = -1.11206 

38 rho = 0 

39 beta_0 = 0.108571 

40 beta_1 = 0.115306 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 lalpha rho beta_0 beta_1 

46 

47 lalpha 1 0.94 0.036 -0.047 

48 

49 rho 0.94 1 0.04 -0.052 

50 

51 beta_0 0.036 0.04 1 -0.46 

52 

53 beta_1 -0.047 -0.052 -0.46 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 95.0% Wald 

60 Confidence Interval 
D-59 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. Lower Conf. Limit 

2 Upper Conf. Limit 

3 lalpha 0.353344 0.480274 -0.587974 

4 1.29466 

5 rho 1.1315 0.292904 0.557421 

6 1.70558 

7 beta_0 0.123135 0.0618608 0.00189039 

8 0.24438 

9 beta_1 0.106469 0.0535364 0.00153987 

10 0.211399 

11 

12 

13 

14 Table of Data and Estimated Values of Interest

15 

16 Dose N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled 

17 Res. 

18 ------ --- -------- -------- ----------- ----------- ---------

19 - 

20 

21 0 28 0.14 0.123 0.37 0.365 0.245 

22 0.7 20 0.15 0.198 0.49 0.477 -0.447 

23 3.5 21 0.52 0.496 0.82 0.802 0.138 

24 

25 

26 

27 Model Descriptions for likelihoods calculated 

28 

29 

30 Model A1: Yij = Mu(i) + e(ij) 

31 Var{e(ij)} = Sigma^2 

32 

33 Model A2: Yij = Mu(i) + e(ij) 

34 Var{e(ij)} = Sigma(i)^2 

35 

36 Model A3: Yij = Mu(i) + e(ij) 

37 Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))

38 Model A3 uses any fixed variance parameters that 

39 were specified by the user 

40 

41 Model R: Yi = Mu + e(i) 

42 Var{e(i)} = Sigma^2 

43 

44 

45 Likelihoods of Interest 

46 

47 Model Log(likelihood) # Param's AIC 

48 A1 5.399546 4 -2.799091 

49 A2 13.307908 6 -14.615816 

50 A3 13.189903 5 -16.379806 

51 fitted 13.167852 4 -18.335705 

52 R 2.264796 2 -0.529591 

53 

54 

55 Explanation of Tests 

56 

57 Test 1: Do responses and/or variances differ among Dose levels? 

58 (A2 vs. R) 

59 Test 2: Are Variances Homogeneous? (A1 vs A2) 

60 Test 3: Are variances adequately modeled? (A2 vs. A3) 
D-60 DRAFT – DO NOT CITE OR QUOTE 

1 Test 4: Does the Model for the Mean Fit? (A3 vs. fitted) 

2 (Note: When rho=0 the results of Test 3 and Test 2 will be the same.) 

3 

4 Tests of Interest 

5 

6 Test -2*log(Likelihood Ratio) Test df p-value 

7 

8 Test 1 22.0862 4 0.0001927 

9 Test 2 15.8167 2 0.0003677 

10 Test 3 0.23601 1 0.6271 

11 Test 4 0.0441012 1 0.8337 

12 

13 The p-value for Test 1 is less than .05. There appears to be a 

14 difference between response and/or variances among the dose levels 

15 It seems appropriate to model the data 

16 

17 The p-value for Test 2 is less than .1. A non-homogeneous variance 

18 model appears to be appropriate 

19 

20 The p-value for Test 3 is greater than .1. The modeled variance appears 

21 to be appropriate here 

22 

23 The p-value for Test 4 is greater than .1. The model chosen seems 

24 to adequately describe the data 

25 

26 

27 Benchmark Dose Computation 

28 

29 Specified effect = 3.56 

30 

31 Risk Type = Point risk 

32 

33 Confidence level = 0.95 

34 

35 BMD = 32.2804 

36 

37 

38 BMDL = 18.094 
D-61 DRAFT – DO NOT CITE OR QUOTE 

1 Nesnow 1998b i.p. multiplicity 

2 BbF 

3 Drop 2 high doses 

4 Linear 

5 Nonconstant variance 

6 BMR = lowest statistically significant response in BaP treated animals (after 

7 control subtracted) 

8 

9 ==================================================================== 

10 Polynomial Model. (Version: 2.12; Date: 02/20/2007) 

11 Input Data File: C:\BMDS\UNSAVED1.(d) 

12 Gnuplot Plotting File: C:\BMDS\UNSAVED1.plt 

13 Mon May 11 20:47:24 2009 

14 ==================================================================== 

15 

16 BMDS MODEL RUN 

17 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

18 

19 The form of the response function is: 

20 

21 Y[dose] = beta_0 + beta_1*dose + beta_2*dose^2 + ... 

22 

23 

24 Dependent variable = MEAN 

25 Independent variable = COLUMN1 

26 The polynomial coefficients are restricted to be positive 

27 The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho) 

28 

29 Total number of dose groups = 3 

30 Total number of records with missing values = 0 

31 Maximum number of iterations = 250 

32 Relative Function Convergence has been set to: 1e-008 

33 Parameter Convergence has been set to: 1e-008 

34 

35 

36 

37 Default Initial Parameter Values 

38 lalpha = 0.21205 

39 rho = 0 

40 beta_0 = 0.453571 

41 beta_1 = 0.0305714 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 lalpha rho beta_0 beta_1 

47 

48 lalpha 1 0.38 0.032 -0.058 

49 

50 rho 0.38 1 -0.032 -0.017 

51 

52 beta_0 0.032 -0.032 1 -0.39 

53 

54 beta_1 -0.058 -0.017 -0.39 1 

55 

56 

57 

58 Parameter Estimates 

59 
D-62 DRAFT – DO NOT CITE OR QUOTE 

1 95.0% Wald 

2 Confidence Interval 

3 Variable Estimate Std. Err. Lower Conf. Limit 

4 Upper Conf. Limit 

5 lalpha 0.158349 0.173664 -0.182027 

6 0.498724 

7 rho 1.42233 0.285984 0.861815 

8 1.98285 

9 beta_0 0.516257 0.101994 0.316353 

10 0.71616 

11 beta_1 0.0272062 0.00781084 0.0118972 

12 0.0425152 

13 

14 

15 

16 Table of Data and Estimated Values of Interest

17 

18 Dose N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled 

19 Res. 

20 ------ --- -------- -------- ----------- ----------- ---------

21 - 

22 

23 0 40 0.525 0.516 0.72 0.676 0.0818 

24 10 18 0.67 0.788 0.75 0.914 -0.549 

25 50 20 2 1.88 1.82 1.69 0.326 

26 

27 

28 

29 Model Descriptions for likelihoods calculated 

30 

31 

32 Model A1: Yij = Mu(i) + e(ij) 

33 Var{e(ij)} = Sigma^2 

34 

35 Model A2: Yij = Mu(i) + e(ij) 

36 Var{e(ij)} = Sigma(i)^2 

37 

38 Model A3: Yij = Mu(i) + e(ij) 

39 Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))

40 Model A3 uses any fixed variance parameters that 

41 were specified by the user 

42 

43 Model R: Yi = Mu + e(i) 

44 Var{e(i)} = Sigma^2 

45 

46 

47 Likelihoods of Interest 

48 

49 Model Log(likelihood) # Param's AIC 

50 A1 -45.740331 4 99.480662 

51 A2 -31.124575 6 74.249150 

52 A3 -31.233847 5 72.467694 

53 fitted -32.276084 4 72.552168 

54 R -56.886387 2 117.772774 

55 

56 

57 Explanation of Tests 

58 

59 Test 1: Do responses and/or variances differ among Dose levels? 

60 (A2 vs. R) 
D-63 DRAFT – DO NOT CITE OR QUOTE 

1 Test 2: Are Variances Homogeneous? (A1 vs A2) 

2 Test 3: Are variances adequately modeled? (A2 vs. A3) 

3 Test 4: Does the Model for the Mean Fit? (A3 vs. fitted) 

4 (Note: When rho=0 the results of Test 3 and Test 2 will be the same.) 

5 

6 Tests of Interest 

7 

8 Test -2*log(Likelihood Ratio) Test df p-value 

9 

10 Test 1 51.5236 4 <.0001 

11 Test 2 29.2315 2 <.0001 

12 Test 3 0.218544 1 0.6402 

13 Test 4 2.08447 1 0.1488 

14 

15 The p-value for Test 1 is less than .05. There appears to be a 

16 difference between response and/or variances among the dose levels 

17 It seems appropriate to model the data 

18 

19 The p-value for Test 2 is less than .1. A non-homogeneous variance 

20 model appears to be appropriate 

21 

22 The p-value for Test 3 is greater than .1. The modeled variance appears 

23 to be appropriate here 

24 

25 The p-value for Test 4 is greater than .1. The model chosen seems 

26 to adequately describe the data 

27 

28 

29 Benchmark Dose Computation 

30 

31 Specified effect = 3.85 

32 

33 Risk Type = Point risk 

34 

35 Confidence level = 0.95 

36 

37 BMD = 122.536 

38 

39 

40 BMDL = 84.4572 
D-64 DRAFT – DO NOT CITE OR QUOTE 

1 Nesnow 1998b i.p. multiplicity 

2 DBahA 

3 Drop 2 high doses 

4 Linear 

5 Nonconstant variance 

6 BMR = lowest statistically significant response in BaP treated animals (after 

7 control subtracted) 

8 

9 ==================================================================== 

10 Polynomial Model. (Version: 2.12; Date: 02/20/2007) 

11 Input Data File: C:\BMDS\UNSAVED1.(d) 

12 Gnuplot Plotting File: C:\BMDS\UNSAVED1.plt 

13 Mon May 11 20:55:01 2009 

14 ==================================================================== 

15 

16 BMDS MODEL RUN 

17 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

18 

19 The form of the response function is: 

20 

21 Y[dose] = beta_0 + beta_1*dose + beta_2*dose^2 + ... 

22 

23 

24 Dependent variable = MEAN 

25 Independent variable = COLUMN1 

26 The polynomial coefficients are restricted to be positive 

27 The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho) 

28 

29 Total number of dose groups = 3 

30 Total number of records with missing values = 0 

31 Maximum number of iterations = 250 

32 Relative Function Convergence has been set to: 1e-008 

33 Parameter Convergence has been set to: 1e-008 

34 

35 

36 

37 Default Initial Parameter Values 

38 lalpha = 0.495312 

39 rho = 0 

40 beta_0 = 0.409167 

41 beta_1 = 1.01 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 lalpha rho beta_0 beta_1 

47 

48 lalpha 1 -0.039 -0.0077 0.0076 

49 

50 rho -0.039 1 0.042 -0.057 

51 

52 beta_0 -0.0077 0.042 1 -0.37 

53 

54 beta_1 0.0076 -0.057 -0.37 1 

55 

56 

57 

58 Parameter Estimates 

59 
D-65 DRAFT – DO NOT CITE OR QUOTE 

1 95.0% Wald 

2 Confidence Interval 

3 Variable Estimate Std. Err. Lower Conf. Limit 

4 Upper Conf. Limit 

5 lalpha 0.090155 0.16129 -0.225967 

6 0.406277 

7 rho 1.13256 0.215446 0.710291 

8 1.55482 

9 beta_0 0.509019 0.111758 0.289977 

10 0.728061 

11 beta_1 0.936099 0.156639 0.629093 

12 1.24311 

13 

14 

15 

16 Table of Data and Estimated Values of Interest

17 

18 Dose N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled 

19 Res. 

20 ------ --- -------- -------- ----------- ----------- ---------

21 - 

22 

23 0 40 0.525 0.509 0.72 0.714 0.142 

24 1.25 18 1.44 1.68 1.46 1.4 -0.723 

25 2.5 19 3.05 2.85 1.9 1.89 0.462 

26 

27 

28 

29 Model Descriptions for likelihoods calculated 

30 

31 

32 Model A1: Yij = Mu(i) + e(ij) 

33 Var{e(ij)} = Sigma^2 

34 

35 Model A2: Yij = Mu(i) + e(ij) 

36 Var{e(ij)} = Sigma(i)^2 

37 

38 Model A3: Yij = Mu(i) + e(ij) 

39 Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))

40 Model A3 uses any fixed variance parameters that 

41 were specified by the user 

42 

43 Model R: Yi = Mu + e(i) 

44 Var{e(i)} = Sigma^2 

45 

46 

47 Likelihoods of Interest 

48 

49 Model Log(likelihood) # Param's AIC 

50 A1 -56.039525 4 120.079049 

51 A2 -42.832497 6 97.664993 

52 A3 -43.013192 5 96.026383 

53 fitted -43.223844 4 94.447689 

54 R -75.955323 2 155.910645 

55 

56 

57 Explanation of Tests 

58 

59 Test 1: Do responses and/or variances differ among Dose levels? 

60 (A2 vs. R) 
D-66 DRAFT – DO NOT CITE OR QUOTE 

1 Test 2: Are Variances Homogeneous? (A1 vs A2) 

2 Test 3: Are variances adequately modeled? (A2 vs. A3) 

3 Test 4: Does the Model for the Mean Fit? (A3 vs. fitted) 

4 (Note: When rho=0 the results of Test 3 and Test 2 will be the same.) 

5 

6 Tests of Interest 

7 

8 Test -2*log(Likelihood Ratio) Test df p-value 

9 

10 Test 1 66.2457 4 <.0001 

11 Test 2 26.4141 2 <.0001 

12 Test 3 0.36139 1 0.5477 

13 Test 4 0.421305 1 0.5163 

14 

15 The p-value for Test 1 is less than .05. There appears to be a 

16 difference between response and/or variances among the dose levels 

17 It seems appropriate to model the data 

18 

19 The p-value for Test 2 is less than .1. A non-homogeneous variance 

20 model appears to be appropriate 

21 

22 The p-value for Test 3 is greater than .1. The modeled variance appears 

23 to be appropriate here 

24 

25 The p-value for Test 4 is greater than .1. The model chosen seems 

26 to adequately describe the data 

27 

28 

29 Benchmark Dose Computation 

30 

31 Specified effect = 3.85 

32 

33 Risk Type = Point risk 

34 

35 Confidence level = 0.95 

36 

37 BMD = 3.56905 

38 

39 

40 BMDL = 2.82758 

41 

42 
D-67 DRAFT – DO NOT CITE OR QUOTE 

1 D.3. LUNG IMPLANTATION BIOASSAYS 

2 DEUTSCH-WENZEL1983AA.OUT.txt 

3 ==================================================================== 

4 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

5 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER ROUTE\DEUTSCH6 WENZEL1983.(d) 

7 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

8 ROUTE\DEUTSCH-WENZEL1983.plt 

9 Fri May 27 10:51:53 2005 

10 ==================================================================== 

11 

12 BMDS MODEL RUN 

13 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

14 

15 The form of the probability function is: 

16 

17 P[response] = background + (1-background)*[1-EXP( 

18 -beta1*dose^1)] 

19 

20 The parameter betas are restricted to be positive 

21 

22 

23 Dependent variable = incidenceAA 

24 Independent variable = doseAA 

25 

26 Total number of observations = 3 

27 Total number of records with missing values = 0 

28 Total number of parameters in model = 2 

29 Total number of specified parameters = 0 

30 Degree of polynomial = 1 

31 

32 

33 Maximum number of iterations = 250 

34 Relative Function Convergence has been set to: 1e-008 

35 Parameter Convergence has been set to: 1e-008 

36 

37 

38 

39 Default Initial Parameter Values 

40 Background = 0 

41 Beta(1) = 0.996523 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 
D-68 DRAFT – DO NOT CITE OR QUOTE 

1 Background 0 NA 

2 Beta(1) 0.773841 0.260605 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -28.6723 

14 Fitted model -30.8245 4.30422 2 0.1162 

15 Reduced model -51.1258 44.907 2 <.0001 

16 

17 AIC: 63.6489 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 35 0.000 

26 i: 2 

27 0.1600 0.1165 4.076 1 35 -0.854 

28 i: 3 

29 0.8300 0.4739 16.587 19 35 0.277 

30 

31 Chi-square = 3.29 DF = 2 P-value = 0.1926 

32 

33 

34 Benchmark Dose Computation 

35 

36 Specified effect = 0.1 

37 

38 Risk Type = Extra risk 

39 

40 Confidence level = 0.95 

41 

42 BMD = 0.136153 

43 

44 BMDL = 0.0956191 
D-69 DRAFT – DO NOT CITE OR QUOTE 

1 DEUTSCH-WENZEL1983BaP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER ROUTE\DEUTSCH5 WENZEL1983.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

7 ROUTE\DEUTSCH-WENZEL1983.plt 

8 Fri May 27 10:42:10 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBaP 

23 Independent variable = doseBaP 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0757681 

40 Beta(1) = 2.82425 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-70 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 3.25323 0.593548 

2 Beta(2) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -51.1075 

14 Fitted model -51.3412 0.467435 3 0.926 

15 Reduced model -96.8119 91.4088 3 <.0001 

16 

17 AIC: 104.682 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 35 0.000 

26 i: 2 

27 0.1000 0.2777 9.720 10 35 0.040 

28 i: 3 

29 0.3000 0.6232 21.811 23 35 0.145 

30 i: 4 

31 1.0000 0.9614 33.647 33 35 -0.498 

32 

33 Chi-square = 0.51 DF = 3 P-value = 0.9177 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 0.1 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 0.0323864 

45 

46 BMDL = 0.0255063 
D-71 DRAFT – DO NOT CITE OR QUOTE 

1 DEUTSCH-WENZEL1983BbF.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER ROUTE\DEUTSCH5 WENZEL1983.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

7 ROUTE\DEUTSCH-WENZEL1983.plt 

8 Fri May 27 10:46:47 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBbF 

23 Independent variable = doseBbF 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.00149382 

40 Beta(1) = 0.226374 

41 Beta(2) = 0.236366 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) Beta(2) 

52 

53 Beta(1) 1 -0.97 

54 

55 Beta(2) -0.97 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-72 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0 NA 

3 Beta(1) 0.24518 0.781411 

4 Beta(2) 0.217701 0.830304 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -37.8686 

16 Fitted model -37.8743 0.0112712 2 0.9944 

17 Reduced model -51.7666 27.796 3 <.0001 

18 

19 AIC: 79.7485 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0000 0.000 0 35 0.000 

28 i: 2 

29 0.1000 0.0263 0.922 1 35 0.087 

30 i: 3 

31 0.3000 0.0889 3.113 3 35 -0.040 

32 i: 4 

33 1.0000 0.3705 12.969 13 35 0.004 

34 

35 Chi-square = 0.01 DF = 2 P-value = 0.9943 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.1 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 0.33191 

47 

48 BMDL = 0.184961 
D-73 DRAFT – DO NOT CITE OR QUOTE 

1 DEUTSCH-WENZEL1983BghiP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER ROUTE\DEUTSCH5 WENZEL1983.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

7 ROUTE\DEUTSCH-WENZEL1983.plt 

8 Fri May 27 10:49:54 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBghiP 

23 Independent variable = doseBghiP 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0.0304801 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) Beta(2) 

52 

53 Beta(1) 1 -0.98 

54 

55 Beta(2) -0.98 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-74 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0 NA 

3 Beta(1) 0.0277423 0.232348 

4 Beta(2) 0.000645059 0.0574865 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -16.8561 

16 Fitted model -17.033 0.353756 2 0.8379 

17 Reduced model -21.5342 9.35614 3 0.02491 

18 

19 AIC: 38.0659 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0000 0.000 0 35 0.000 

28 i: 2 

29 0.1600 0.0044 0.156 0 35 -1.004 

30 i: 3 

31 0.8300 0.0232 0.812 1 35 0.237 

32 i: 4 

33 4.1500 0.1186 4.032 4 34 -0.009 

34 

35 Chi-square = 0.20 DF = 2 P-value = 0.9043 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.1 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 3.51117 

47 

48 BMDL = 1.82558 
D-75 DRAFT – DO NOT CITE OR QUOTE 

1 DEUTSCH-WENZEL1983BjF.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER ROUTE\DEUTSCH5 WENZEL1983.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

7 ROUTE\DEUTSCH-WENZEL1983.plt 

8 Fri May 27 10:47:32 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBjF 

23 Independent variable = doseBjF 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.00616121 

40 Beta(1) = 0.0709095 

41 Beta(2) = 0.0144537 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) Beta(2) 

52 

53 Beta(1) 1 -0.98 

54 

55 Beta(2) -0.98 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-76 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0 NA 

3 Beta(1) 0.0929144 0.226076 

4 Beta(2) 0.0101278 0.0466964 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -39.0246 

16 Fitted model -39.1336 0.218103 2 0.8967 

17 Reduced model -60.8862 43.7233 3 <.0001 

18 

19 AIC: 82.2673 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0000 0.000 0 35 0.000 

28 i: 2 

29 0.2000 0.0188 0.658 1 35 0.529 

30 i: 3 

31 1.0000 0.0979 3.427 3 35 -0.138 

32 i: 4 

33 5.0000 0.5122 17.926 18 35 0.009 

34 

35 Chi-square = 0.24 DF = 2 P-value = 0.8868 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.1 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 1.02045 

47 

48 BMDL = 0.580958 
D-77 DRAFT – DO NOT CITE OR QUOTE 

1 DEUTSCH-WENZEL1983BkF.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER ROUTE\DEUTSCH5 WENZEL1983.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

7 ROUTE\DEUTSCH-WENZEL1983.plt 

8 Fri May 27 10:48:11 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceBkF 

23 Independent variable = doseBkF 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0.126747 

41 Beta(2) = 0.00410997 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) Beta(2) 

52 

53 Beta(1) 1 -0.97 

54 

55 Beta(2) -0.97 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-78 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0 NA 

3 Beta(1) 0.0842968 0.251118 

4 Beta(2) 0.0142917 0.0632842 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -28.404 

16 Fitted model -28.9719 1.1357 2 0.5667 

17 Reduced model -46.2443 35.6806 3 <.0001 

18 

19 AIC: 61.9437 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0000 0.000 0 35 0.000 

28 i: 2 

29 0.1600 0.0138 0.482 0 35 -1.014 

30 i: 3 

31 0.8300 0.0767 2.378 3 31 0.283 

32 i: 4 

33 4.1500 0.4490 12.122 12 27 -0.018 

34 

35 Chi-square = 0.67 DF = 2 P-value = 0.7165 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.1 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 1.05954 

47 

48 BMDL = 0.557079 
D-79 DRAFT – DO NOT CITE OR QUOTE 

1 DEUTSCH-WENZEL1983IP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER ROUTE\DEUTSCH5 WENZEL1983.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

7 ROUTE\DEUTSCH-WENZEL1983.plt 

8 Fri May 27 10:49:04 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = incidenceIP 

23 Independent variable = doseIP 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0539703 

40 Beta(1) = 0.20919 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Background Beta(1) 

52 

53 Background 1 -0.55 

54 

55 Beta(1) -0.55 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-80 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0.0224449 0.113638 

3 Beta(1) 0.241452 0.0797033 

4 Beta(2) 0 NA 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -54.8079 

16 Fitted model -56.5662 3.5166 2 0.1723 

17 Reduced model -76.4525 43.2893 3 <.0001 

18 

19 AIC: 117.132 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0224 0.786 0 35 -1.023 

28 i: 2 

29 0.1600 0.0595 2.082 4 35 0.979 

30 i: 3 

31 0.8300 0.2000 6.999 8 35 0.179 

32 i: 4 

33 4.1500 0.6411 22.439 21 35 -0.179 

34 

35 Chi-square = 3.12 DF = 2 P-value = 0.2104 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.1 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 0.436361 

47 

48 BMDL = 0.309504 
D-81 DRAFT – DO NOT CITE OR QUOTE 

1 WENZEL-HARTUNG1990BaP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER ROUTE\WENZEL5 HARTUNG1990.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

7 ROUTE\WENZEL-HARTUNG1990.plt 

8 Fri May 27 10:58:05 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = responseBaP 

23 Independent variable = doseBaP 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 3.21631 

41 Beta(2) = 5.7325 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) Beta(2) 

52 

53 Beta(1) 1 -0.93 

54 

55 Beta(2) -0.93 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-82 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0 NA 

3 Beta(1) 3.01149 2.79594 

4 Beta(2) 6.44644 10.7674 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -50.8389 

16 Fitted model -50.8521 0.0264626 2 0.9869 

17 Reduced model -84.6566 67.6355 3 <.0001 

18 

19 AIC: 105.704 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0000 0.000 0 35 0.000 

28 i: 2 

29 0.0300 0.0917 3.208 3 35 -0.072 

30 i: 3 

31 0.1000 0.3062 10.718 11 35 0.038 

32 i: 4 

33 0.3000 0.7732 27.062 27 35 -0.010 

34 

35 Chi-square = 0.03 DF = 2 P-value = 0.9870 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.1 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 0.0326976 

47 

48 BMDL = 0.0198862 
D-83 DRAFT – DO NOT CITE OR QUOTE 

1 WENZEL-HARTUNG1990BaPforDBahA.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

5 ROUTE\SETS\WENZEL-HARTUNG1990.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

7 ROUTE\SETS\WENZEL-HARTUNG1990.plt 

8 Thu Jun 02 09:02:58 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = responseBaP 

23 Independent variable = doseBaP 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 3.21631 

41 Beta(2) = 5.7325 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) Beta(2) 

52 

53 Beta(1) 1 -0.93 

54 

55 Beta(2) -0.93 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-84 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0 NA 

3 Beta(1) 3.01149 2.79594 

4 Beta(2) 6.44644 10.7674 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -50.8389 

16 Fitted model -50.8521 0.0264626 2 0.9869 

17 Reduced model -84.6566 67.6355 3 <.0001 

18 

19 AIC: 105.704 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0000 0.000 0 35 0.000 

28 i: 2 

29 0.0300 0.0917 3.208 3 35 -0.072 

30 i: 3 

31 0.1000 0.3062 10.718 11 35 0.038 

32 i: 4 

33 0.3000 0.7732 27.062 27 35 -0.010 

34 

35 Chi-square = 0.03 DF = 2 P-value = 0.9870 

36 

37 

38 Benchmark Dose Computation 

39 

40 Specified effect = 0.57 

41 

42 Risk Type = Extra risk 

43 

44 Confidence level = 0.95 

45 

46 BMD = 0.197095 

47 

48 BMDL = 0.157781 
D-85 DRAFT – DO NOT CITE OR QUOTE 

1 WENZEL-HARTUNG1990CH.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER ROUTE\WENZEL5 HARTUNG1990.(d) 

6 Gnuplot Plotting File: C:\PAH\BMD ANALYSIS\BIOASSAY\OTHER 

7 ROUTE\WENZEL-HARTUNG1990.plt 

8 Fri May 27 10:58:53 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = responseCH 

23 Independent variable = doseCH 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0178361 

40 Beta(1) = 0.109158 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 ( *** The model parameter(s) -Background 

46 have been estimated at a boundary point, or have been 

47 specified by the user, 

48 and do not appear in the correlation matrix ) 

49 

50 Beta(1) 

51 

52 Beta(1) 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0 NA 

60 Beta(1) 0.123432 0.0647008 
D-86 DRAFT – DO NOT CITE OR QUOTE 

1 

2 NA - Indicates that this parameter has hit a bound 

3 implied by some inequality constraint and thus

4 has no standard error. 

5 

6 

7 

8 Analysis of Deviance Table 

9 

10 Model Log(likelihood) Deviance Test DF P-value 

11 Full model -35.2935 

12 Fitted model -35.455 0.323044 2 0.8508 

13 Reduced model -43.0622 15.5374 2 0.0004228 

14 

15 AIC: 72.9101 

16 

17 

18 Goodness of Fit 

19 

20 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

21 ----------------------------------------------------------------------- 

22 i: 1 

23 0.0000 0.0000 0.000 0 35 0.000 

24 i: 2 

25 1.0000 0.1161 4.064 5 35 0.261 

26 i: 3 

27 3.0000 0.3095 10.831 10 35 -0.111 

28 

29 Chi-square = 0.34 DF = 2 P-value = 0.8453 

30 

31 

32 Benchmark Dose Computation 

33 

34 Specified effect = 0.1 

35 

36 Risk Type = Extra risk 

37 

38 Confidence level = 0.95 

39 

40 BMD = 0.853595 

41 

42 BMDL = 0.57298 

43 
D-87 DRAFT – DO NOT CITE OR QUOTE 

1 D.4. BACTERIAL MUTAGENICITY 

2 Hass 1981 bact mut bap.out.txt 

3 ==================================================================== 

4 Polynomial Model. Revision: 2.2 Date: 9/12/2002 

5 Input Data File: C:\BMDS\UNSAVED1.(d) 

6 Gnuplot Plotting File: C:\BMDS\UNSAVED1.plt 

7 Wed Jul 06 11:29:07 2005 

8 ==================================================================== 

9 

10 BMDS MODEL RUN 

11 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

12 

13 The form of the response function is: 

14 

15 Y[dose] = beta_0 + beta_1*dose + beta_2*dose^2 + ... 

16 

17 

18 Dependent variable = MEAN 

19 Independent variable = COLUMN1 

20 rho is set to 0 

21 Signs of the polynomial coefficients are not restricted 

22 A constant variance model is fit 

23 

24 Total number of dose groups = 4 

25 Total number of records with missing values = 0 

26 Maximum number of iterations = 250 

27 Relative Function Convergence has been set to: 1e-008 

28 Parameter Convergence has been set to: 1e-008 

29 

30 

31 

32 Default Initial Parameter Values 

33 alpha = 194.5 

34 rho = 0 Specified 

35 beta_0 = 121.8 

36 beta_1 = 297.029 

37 

38 

39 

40 Parameter Estimates 

41 

42 95.0% Wald 

43 Confidence Interval 

44 Variable Estimate Std. Err. Lower Conf. Limit 

45 Upper Conf. Limit 

46 alpha 132.71 54.1784 26.5217 

47 238.897 

48 beta_0 121.8 5.15188 111.702 

49 131.898 

50 beta_1 297.029 8.99387 279.401 

51 314.656 

52 

53 

54 Asymptotic Correlation Matrix of Parameter Estimates 

55 

56 alpha beta_0 beta_1 

57 alpha 1 -1.4e-009 -1.1e-008 

58 beta_0 -1.4e-009 1 -0.76 

59 beta_1 -1.1e-008 -0.76 1 
D-88 DRAFT – DO NOT CITE OR QUOTE 

1 

2 

3 Table of Data and Estimated Values of Interest

4 

5 Dose N Obs Mean Obs Std Dev Est Mean Est Std Dev Chi^2 

6 Res. 

7 ------ --- -------- ----------- -------- ----------- ---------

8 - 

9 

10 0 3 124 8 122 11.5 0.331 

11 0.25 3 194 16 196 11.5 -0.309 

12 0.5 3 269 13 270 11.5 -0.198 

13 1 3 420 17 419 11.5 0.176 

14 

15 

16 

17 Model Descriptions for likelihoods calculated 

18 

19 

20 Model A1: Yij = Mu(i) + e(ij) 

21 Var{e(ij)} = Sigma^2 

22 

23 Model A2: Yij = Mu(i) + e(ij) 

24 Var{e(ij)} = Sigma(i)^2 

25 

26 Model R: Yi = Mu + e(i) 

27 Var{e(i)} = Sigma^2 

28 

29 

30 Likelihoods of Interest 

31 

32 Model Log(likelihood) DF AIC 

33 A1 -35.189802 5 80.379605 

34 A2 -34.317788 8 84.635576 

35 fitted -35.328976 2 74.657952 

36 R -62.974684 2 129.949369 

37 

38 Test 1: Does response and/or variances differ among dose 

39 levels 

40 (A2 vs. R) 

41 Test 2: Are Variances Homogeneous (A1 vs A2) 

42 Test 3: Does the Model for the Mean Fit (A1 vs. fitted) 

43 

44 Tests of Interest 

45 

46 Test -2*log(Likelihood Ratio) Test df p-value 

47 

48 Test 1 57.3138 6 <.0001 

49 Test 2 1.74403 3 0.6272 

50 Test 3 0.278348 2 0.8701 

51 

52 The p-value for Test 1 is less than .05. There appears 

53 to be a 

54 difference between response and/or variances among the 

55 dose levels. 

56 It seems appropriate to model the data 

57 

58 The p-value for Test 2 is greater than .05. A 

59 homogeneous variance 

60 model appears to be appropriate here 
D-89 DRAFT – DO NOT CITE OR QUOTE 

1 

2 

3 The p-value for Test 3 is greater than .05. The model 

4 chosen appears 

5 to adequately describe the data 

6 

7 

8 

9 Benchmark Dose Computation 

10 Specified effect = 1 

11 

12 Risk Type = Estimated standard deviations from the control mean 

13 

14 

15 Confidence level = 0.95 

16 

17 BMD = 0.038784 

18 

19 

20 BMDL = 0.0286028 

21 
D-90 DRAFT – DO NOT CITE OR QUOTE 

1 HASS_1981_BACT_MUT_BEP.OUT.txt 

2 ==================================================================== 

3 Polynomial Model. Revision: 2.2 Date: 9/12/2002 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\HASS_1981_BACT_MUT_BEP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\HASS_1981_BACT_MUT_BEP.plt 

8 Wed Jul 06 13:42:38 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the response function is: 

15 

16 Y[dose] = beta_0 + beta_1*dose + beta_2*dose^2 + ... 

17 

18 

19 Dependent variable = MEAN 

20 Independent variable = COLUMN1 

21 rho is set to 0 

22 Signs of the polynomial coefficients are not restricted 

23 A constant variance model is fit 

24 

25 Total number of dose groups = 4 

26 Total number of records with missing values = 0 

27 Maximum number of iterations = 250 

28 Relative Function Convergence has been set to: 1e-008 

29 Parameter Convergence has been set to: 1e-008 

30 

31 

32 

33 Default Initial Parameter Values 

34 alpha = 117.5 

35 rho = 0 Specified 

36 beta_0 = 120.75 

37 beta_1 = 77.5 

38 

39 

40 

41 Parameter Estimates 

42 

43 95.0% Wald 

44 Confidence Interval 

45 Variable Estimate Std. Err. Lower Conf. Limit 

46 Upper Conf. Limit 

47 alpha 98.6458 40.272 19.7142 

48 177.577 

49 beta_0 120.75 4.19706 112.524 

50 128.976 

51 beta_1 77.5 7.66275 62.4813 

52 92.5187 

53 

54 

55 Asymptotic Correlation Matrix of Parameter Estimates 

56 

57 alpha beta_0 beta_1 

58 alpha 1 -8e-012 1.1e-011 

59 beta_0 -8e-012 1 -0.73 

60 beta_1 1.1e-011 -0.73 1 
D-91 DRAFT – DO NOT CITE OR QUOTE 

1 

2 

3 Table of Data and Estimated Values of Interest

4 

5 Dose N Obs Mean Obs Std Dev Est Mean Est Std Dev Chi^2 

6 Res. 

7 ------ --- -------- ----------- -------- ----------- ---------

8 - 

9 

10 0 3 124 8 121 9.93 0.567 

11 0.2 3 129 6 136 9.93 -1.26 

12 0.4 3 156 9 152 9.93 0.741 

13 1 3 198 17 198 9.93 -0.0436 

14 

15 

16 

17 Model Descriptions for likelihoods calculated 

18 

19 

20 Model A1: Yij = Mu(i) + e(ij) 

21 Var{e(ij)} = Sigma^2 

22 

23 Model A2: Yij = Mu(i) + e(ij) 

24 Var{e(ij)} = Sigma(i)^2 

25 

26 Model R: Yi = Mu + e(i) 

27 Var{e(i)} = Sigma^2 

28 

29 

30 Likelihoods of Interest 

31 

32 Model Log(likelihood) DF AIC 

33 A1 -32.165839 5 74.331679 

34 A2 -30.272126 8 76.544252 

35 fitted -33.549216 2 71.098432 

36 R -47.594288 2 99.188576 

37 

38 Test 1: Does response and/or variances differ among dose 

39 levels 

40 (A2 vs. R) 

41 Test 2: Are Variances Homogeneous (A1 vs A2) 

42 Test 3: Does the Model for the Mean Fit (A1 vs. fitted) 

43 

44 Tests of Interest 

45 

46 Test -2*log(Likelihood Ratio) Test df p-value 

47 

48 Test 1 34.6443 6 <.0001 

49 Test 2 3.78743 3 0.2854 

50 Test 3 2.76675 2 0.2507 

51 

52 The p-value for Test 1 is less than .05. There appears 

53 to be a 

54 difference between response and/or variances among the 

55 dose levels. 

56 It seems appropriate to model the data 

57 

58 The p-value for Test 2 is greater than .05. A 

59 homogeneous variance 

60 model appears to be appropriate here 
D-92 DRAFT – DO NOT CITE OR QUOTE 

1 

2 

3 The p-value for Test 3 is greater than .05. The model 

4 chosen appears 

5 to adequately describe the data 

6 

7 

8 

9 Benchmark Dose Computation 

10 Specified effect = 1 

11 

12 Risk Type = Estimated standard deviations from the control mean 

13 

14 

15 Confidence level = 0.95 

16 

17 BMD = 0.128156 

18 

19 

20 BMDL = 0.0923937 

21 
D-93 DRAFT – DO NOT CITE OR QUOTE 

1 JOHNSEN_1997_BAC_MUT_BAP.OUT.txt 

2 ==================================================================== 

3 Polynomial Model. Revision: 2.2 Date: 9/12/2002 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\JOHNSEN_1997_BAC_MUT_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\JOHNSEN_1997_BAC_MUT_BAP.plt 

8 Fri Jul 08 09:02:29 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the response function is: 

15 

16 Y[dose] = beta_0 + beta_1*dose + beta_2*dose^2 + ... 

17 

18 

19 Dependent variable = MEAN 

20 Independent variable = COLUMN1 

21 rho is set to 0 

22 Signs of the polynomial coefficients are not restricted 

23 A constant variance model is fit 

24 

25 Total number of dose groups = 3 

26 Total number of records with missing values = 0 

27 Maximum number of iterations = 250 

28 Relative Function Convergence has been set to: 1e-008 

29 Parameter Convergence has been set to: 1e-008 

30 

31 

32 

33 Default Initial Parameter Values 

34 alpha = 70.2768 

35 rho = 0 Specified 

36 beta_0 = 115.5 

37 beta_1 = 0.65 

38 

39 

40 

41 Parameter Estimates 

42 

43 95.0% Wald 

44 Confidence Interval 

45 Variable Estimate Std. Err. Lower Conf. Limit 

46 Upper Conf. Limit 

47 alpha 59.3512 27.9784 4.51449 

48 114.188 

49 beta_0 115.5 4.06035 107.542 

50 123.458 

51 beta_1 0.65 0.314513 0.0335651 

52 1.26643 

53 

54 

55 Asymptotic Correlation Matrix of Parameter Estimates 

56 

57 alpha beta_0 beta_1 

58 alpha 1 -7.9e-010 -3.4e-012 

59 beta_0 -7.9e-010 1 -0.77 

60 beta_1 -3.4e-012 -0.77 1 
D-94 DRAFT – DO NOT CITE OR QUOTE 

1 

2 

3 Table of Data and Estimated Values of Interest

4 

5 Dose N Obs Mean Obs Std Dev Est Mean Est Std Dev Chi^2 

6 Res. 

7 ------ --- -------- ----------- -------- ----------- ---------

8 - 

9 

10 0 3 113 9.68 115 7.7 -0.562 

11 10 3 127 4.84 122 7.7 1.12 

12 20 3 126 9.68 128 7.7 -0.562 

13 

14 

15 

16 Model Descriptions for likelihoods calculated 

17 

18 

19 Model A1: Yij = Mu(i) + e(ij) 

20 Var{e(ij)} = Sigma^2 

21 

22 Model A2: Yij = Mu(i) + e(ij) 

23 Var{e(ij)} = Sigma(i)^2 

24 

25 Model R: Yi = Mu + e(i) 

26 Var{e(i)} = Sigma^2 

27 

28 

29 Likelihoods of Interest 

30 

31 Model Log(likelihood) DF AIC 

32 A1 -21.811395 4 51.622790 

33 A2 -21.026523 6 54.053045 

34 fitted -22.875626 2 49.751251 

35 R -24.653317 2 53.306634 

36 

37 Test 1: Does response and/or variances differ among dose 

38 levels 

39 (A2 vs. R) 

40 Test 2: Are Variances Homogeneous (A1 vs A2) 

41 Test 3: Does the Model for the Mean Fit (A1 vs. fitted) 

42 

43 Tests of Interest 

44 

45 Test -2*log(Likelihood Ratio) Test df p-value 

46 

47 Test 1 7.25359 4 0.0266 

48 Test 2 1.56974 2 0.4562 

49 Test 3 2.12846 1 0.1446 

50 

51 The p-value for Test 1 is less than .05. There appears 

52 to be a 

53 difference between response and/or variances among the 

54 dose levels. 

55 It seems appropriate to model the data 

56 

57 The p-value for Test 2 is greater than .05. A 

58 homogeneous variance 

59 model appears to be appropriate here 

60 
D-95 DRAFT – DO NOT CITE OR QUOTE 

1 

2 The p-value for Test 3 is greater than .05. The model 

3 chosen appears 

4 to adequately describe the data 

5 

6 

7 

8 Benchmark Dose Computation 

9 Specified effect = 1 

10 

11 Risk Type = Estimated standard deviations from the control mean 

12 

13 

14 Confidence level = 0.95 

15 

16 BMD = 11.8523 

17 

18 

19 BMDL = 6.27094 

20 

21 

22 
D-96 DRAFT – DO NOT CITE OR QUOTE 

1 D.5. MAMMALIAN MUTAGENICITY 

2 BARF_MUT_BAA.OUT.txt 

3 ==================================================================== 

4 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

5 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

6 RPS\MODELING\BARF_MUT_BAA.(d) 

7 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

8 DOCUMENTS\PAH RPS\MODELING\BARF_MUT_BAA.plt 

9 Thu Jun 30 12:46:38 2005 

10 ==================================================================== 

11 

12 BMDS MODEL RUN 

13 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

14 

15 The form of the probability function is: 

16 

17 P[response] = background + (1-background)*[1-EXP( 

18 -beta1*dose^1-beta2*dose^2-beta3*dose^3)] 

19 

20 The parameter betas are restricted to be positive 

21 

22 

23 Dependent variable = COLUMN2 

24 Independent variable = COLUMN1 

25 

26 Total number of observations = 5 

27 Total number of records with missing values = 0 

28 Total number of parameters in model = 4 

29 Total number of specified parameters = 0 

30 Degree of polynomial = 3 

31 

32 

33 Maximum number of iterations = 250 

34 Relative Function Convergence has been set to: 1e-008 

35 Parameter Convergence has been set to: 1e-008 

36 

37 

38 

39 Default Initial Parameter Values 

40 Background = 3.89426e-006 

41 Beta(1) = 3.46216e-007 

42 Beta(2) = 0 

43 Beta(3) = 1.93939e-012 

44 **** WARNING: Completion code = -2. Optimum not found. Trying new starting 

45 pont  

46 

47 

48 

49 Asymptotic Correlation Matrix of Parameter Estimates 

50 

51 ( ** * The model parameter(s) -Background -Beta(2) -Beta(3) 

52 have been estimated at a boundary point, or have been 

53 specified by the user, 

54 and do not appear in the correlation matrix ) 

55 

56 Beta(1) 

57 

58 Beta(1) 1 

59 
D-97 DRAFT – DO NOT CITE OR QUOTE 

1 

2 

3 Parameter Estimates 

4 

5 Variable Estimate Std. Err. 

6 Background 0 NA 

7 Beta(1) 4.34385e-007 5.43792e-006 

8 Beta(2) 0 NA 

9 Beta(3) 0 NA 

10 

11 NA - Indicates that this parameter has hit a bound 

12 implied by some inequality constraint and thus

13 has no standard error. 

14 

15 

16 

17 Analysis of Deviance Table 

18 

19 Model Log(likelihood) Deviance Test DF P-value 

20 Full model -1545.82 

21 Fitted model -1548.6 5.57201 4 0.2335 

22 Reduced model -1597.17 102.713 4 <.0001 

23 

24 AIC: 3099.21 

25 

26 

27 Goodness of Fit 

28 

29 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

30 ----------------------------------------------------------------------- 

31 i: 1 

32 0.0000 0.0000 0.000 0 1000000 0.000 

33 i: 2 

34 20.0000 0.0000 8.688 12 1000000 0.381 

35 i: 3 

36 50.0000 0.0000 21.719 29 1000000 0.335 

37 i: 4 

38 100.0000 0.0000 43.438 34 1000000 -0.217 

39 i: 5 

40 150.0000 0.0001 65.156 64 1000000 -0.018 

41 

42 Chi-square = 5.77 DF = 4 P-value = 0.2166 

43 

44 

45 Benchmark Dose Computation 

46 

47 Specified effect = 1e-005 

48 

49 Risk Type = Extra risk 

50 

51 Confidence level = 0.95 

52 

53 BMD = 23.0212 

54 

55 **** WARNING: Completion code = -2. Optimum not found. Trying new starting 

56 point  

57 

58 ** ** WARNING 0: Completion code = -2 trying new start  

59 

60 ** ** WARNING 1: Completion code = -2 trying new start  
D-98 DRAFT – DO NOT CITE OR QUOTE 

1 

2 ** ** WARNING 2: Completion code = -2 trying new start  

3 

4 ** ** WARNING 3: Completion code = -2 trying new start  

5 

6 ** ** WARNING 4: Completion code = -2 trying new start  

7 

8 ** ** WARNING 5: Completion code = -2 trying new start  

9 

10 ** ** WARNING 6: Completion code = -2 trying new start  

11 

12 ** ** WARNING 7: Completion code = -2 trying new start  

13 

14 ** ** WARNING 8: Completion code = -2 trying new start  

15 

16 ** ** WARNING 9: Completion code = -2 trying new start  

17 

18 ** ** WARNING: Completion code = -2. Optimum not found. Trying new starting 

19 point  

20 

21 ** ** WARNING 0: Completion code = -2 trying new start  

22 

23 ** ** WARNING 1: Completion code = -3 trying new start  

24 

25 ** ** WARNING 2: Completion code = -3 trying new start  

26 

27 ** ** WARNING 3: Completion code = -3 trying new start  

28 

29 ** ** WARNING 4: Completion code = -3 trying new start  

30 

31 ** ** WARNING 5: Completion code = -3 trying new start  

32 

33 ** ** WARNING 6: Completion code = -2 trying new start  

34 

35 ** ** WARNING 7: Completion code = -3 trying new start  

36 

37 ** ** WARNING 8: Completion code = -3 trying new start  

38 

39 ** ** WARNING 9: Completion code = -3 trying new start**** 

40 

41 

42 Warning: completion code still negative 

43 BMDL did not converge for BMR = 0.000010 

44 

45 Program execution is stopped 
D-99 DRAFT – DO NOT CITE OR QUOTE 

1 BARF_MUT_BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\BARF_MUT_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\BARF_MUT_BAP.plt 

8 Thu Jun 30 12:40:17 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 1.39884e-006 

40 Beta(1) = 5.34042e-006 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-100 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 5.43367e-006 2.68102e-005 

2 Beta(2) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -3273.08 

14 Fitted model -3273.96 1.75092 3 0.6257 

15 Reduced model -3395.25 244.327 3 <.0001 

16 

17 AIC: 6549.92 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 1000000 0.000 

26 i: 2 

27 10.0000 0.0001 54.335 51 1000000 -0.061 

28 i: 3 

29 20.0000 0.0001 108.668 120 1000000 0.104 

30 i: 4 

31 30.0000 0.0002 162.997 155 1000000 -0.049 

32 

33 Chi-square = 1.78 DF = 3 P-value = 0.6195 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 1e-005 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 1.84039 

45 

46   WARNING: Completion code = -3. Optimum not found. Trying new starting 

47 point** ** 

48 

49   WARNING 0: Completion code = -3 trying new start** ** 

50 

51   WARNING 1: Completion code = -3 trying new start** ** 

52 

53   WARNING 2: Completion code = -3 trying new start** ** 

54 

55   WARNING 3: Completion code = -3 trying new start** ** 

56 

57   WARNING 4: Completion code = -3 trying new start** ** 

58 

59   WARNING 5: Completion code = -3 trying new start** ** 

60 
D-101 DRAFT – DO NOT CITE OR QUOTE 

1   WARNING 6: Completion code = -3 trying new start** ** 

2 

3   WARNING 7: Completion code = -3 trying new start** ** 

4 

5   WARNING 8: Completion code = -3 trying new start** ** 

6 

7   WARNING 9: Completion code = -3 trying new start** ** 

8 

9   WARNING: Completion code = -3. Optimum not found. Trying new starting 

10 point** ** 

11 

12   WARNING 0: Completion code = -1 trying new start** ** 

13 

14   WARNING 1: Completion code = -1 trying new start** ** 

15 

16   WARNING 2: Completion code = -1 trying new start** ** 

17 

18 BMDL = 1.68248 
D-102 DRAFT – DO NOT CITE OR QUOTE 

1 BARF_MUT_CH.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\BARF_MUT_CH.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\BARF_MUT_CH.plt 

8 Thu Jun 30 12:48:57 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 2.60526e-006 

40 Beta(1) = 5.02638e-007 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 ( *** The model parameter(s) -Background 

46 have been estimated at a boundary point, or have been 

47 specified by the user, 

48 and do not appear in the correlation matrix ) 

49 

50 Beta(1) 

51 

52 Beta(1) 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0 NA 

60 Beta(1) 6.14293e-007 1.93539e-005 
D-103 DRAFT – DO NOT CITE OR QUOTE 

1 

2 NA - Indicates that this parameter has hit a bound 

3 implied by some inequality constraint and thus

4 has no standard error. 

5 

6 

7 

8 Analysis of Deviance Table 

9 

10 Model Log(likelihood) Deviance Test DF P-value 

11 Full model -504.191 

12 Fitted model -505.38 2.37752 2 0.3046 

13 Reduced model -522.575 36.7681 2 <.0001 

14 

15 AIC: 1012.76 

16 

17 

18 Goodness of Fit 

19 

20 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

21 ----------------------------------------------------------------------- 

22 i: 1 

23 0.0000 0.0000 0.000 0 1000000 0.000 

24 i: 2 

25 20.0000 0.0000 12.286 17 1000000 0.384 

26 i: 3 

27 50.0000 0.0000 30.714 26 1000000 -0.153 

28 

29 Chi-square = 2.53 DF = 2 P-value = 0.2819 

30 

31 

32 Benchmark Dose Computation 

33 

34 Specified effect = 1e-005 

35 

36 Risk Type = Extra risk 

37 

38 Confidence level = 0.95 

39 

40 BMD = 16.279 

41 

42   WARNING: Completion code = -1. Optimum not found. Trying new starting 

43 point** ** 

44 

45   WARNING 0: Completion code = -1 trying new start** ** 

46 

47   WARNING 1: Completion code = -1 trying new start** ** 

48 

49   WARNING 2: Completion code = -1 trying new start** ** 

50 

51   WARNING 3: Completion code = -1 trying new start** ** 

52 

53   WARNING 4: Completion code = -1 trying new start** ** 

54 

55   WARNING 5: Completion code = -1 trying new start** ** 

56 

57   WARNING 6: Completion code = -1 trying new start** ** 

58 

59   WARNING 7: Completion code = -1 trying new start** ** 

60 
D-104 DRAFT – DO NOT CITE OR QUOTE 

1   WARNING 8: Completion code = -1 trying new start** ** 

2 

3   WARNING 9: Completion code = -1 trying new start** ** 

4 

5   WARNING: Completion code = -1. Optimum not found. Trying new starting 

6 point** ** 

7 

8   WARNING 0: Completion code = -3 trying new start** ** 

9 

10   WARNING 1: Completion code = -3 trying new start** ** 

11 

12   WARNING 2: Completion code = -3 trying new start** ** 

13 

14   WARNING 3: Completion code = -3 trying new start** ** 

15 

16   WARNING 4: Completion code = -3 trying new start** ** 

17 

18   WARNING 5: Completion code = -3 trying new start** ** 

19 

20   WARNING 6: Completion code = -3 trying new start** ** 

21 

22   WARNING 7: Completion code = -3 trying new start** ** 

23 

24   WARNING 8: Completion code = -3 trying new start** ** 

25 

26   WARNING 9: Completion code = -3 trying new start** ** 

27 

28 

29 Warning: completion code still negative 

30 BMDL did not converge for BMR = 0.000010 

31 

32 Program execution is stopped 
D-105 DRAFT – DO NOT CITE OR QUOTE 

1 BARF_MUT_FA.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\BARF_MUT_FA.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\BARF_MUT_FA.plt 

8 Thu Jun 30 12:43:11 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 6.6658e-007 

40 Beta(1) = 2.50006e-006 

41 

42 

43 Asymptotic Correlation Matrix of Parameter Estimates 

44 

45 ( *** The model parameter(s) -Background 

46 have been estimated at a boundary point, or have been 

47 specified by the user, 

48 and do not appear in the correlation matrix ) 

49 

50 Beta(1) 

51 

52 Beta(1) 1 

53 

54 

55 

56 Parameter Estimates 

57 

58 Variable Estimate Std. Err. 

59 Background 0 NA 

60 Beta(1) 2.56672e-006 4.49565e-005 
D-106 DRAFT – DO NOT CITE OR QUOTE 

1 

2 NA - Indicates that this parameter has hit a bound 

3 implied by some inequality constraint and thus

4 has no standard error. 

5 

6 

7 

8 Analysis of Deviance Table 

9 

10 Model Log(likelihood) Deviance Test DF P-value 

11 Full model -856.204 

12 Fitted model -856.255 0.103 2 0.9498 

13 Reduced model -890.913 69.419 2 <.0001 

14 

15 AIC: 1714.51 

16 

17 

18 Goodness of Fit 

19 

20 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

21 ----------------------------------------------------------------------- 

22 i: 1 

23 0.0000 0.0000 0.000 0 1000000 0.000 

24 i: 2 

25 10.0000 0.0000 25.667 27 1000000 0.052 

26 i: 3 

27 20.0000 0.0001 51.333 50 1000000 -0.026 

28 

29 Chi-square = 0.10 DF = 2 P-value = 0.9494 

30 

31 

32 Benchmark Dose Computation 

33 

34 Specified effect = 1e-005 

35 

36 Risk Type = Extra risk 

37 

38 Confidence level = 0.95 

39 

40 BMD = 3.89604 

41 

42   WARNING: Completion code = -1. Optimum not found. Trying new starting 

43 point** ** 

44 

45   WARNING 0: Completion code = -1 trying new start** ** 

46 

47   WARNING 1: Completion code = -5 trying new start** ** 

48 

49 BMDL = 0 

50 
D-107 DRAFT – DO NOT CITE OR QUOTE 

1 BARF_MUT_TPHEN.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\BARF_MUT_TPHEN.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\BARF_MUT_TPHEN.plt 

8 Thu Jun 30 12:52:56 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 9.99937e-007 

40 Beta(1) = 1.74289e-007 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-108 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 1.85717e-007 4.42148e-006 

2 Beta(2) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -755.63 

14 Fitted model -755.773 0.2868 3 0.9625 

15 Reduced model -781.782 52.3039 3 <.0001 

16 

17 AIC: 1513.55 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 1000000 0.000 

26 i: 2 

27 50.0000 0.0000 9.286 10 1000000 0.077 

28 i: 3 

29 100.0000 0.0000 18.572 20 1000000 0.077 

30 i: 4 

31 200.0000 0.0000 37.143 35 1000000 -0.058 

32 

33 Chi-square = 0.29 DF = 3 P-value = 0.9622 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 1e-005 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 53.8457 

45 

46   WARNING: Completion code = -2. Optimum not found. Trying new starting 

47 point** ** 

48 

49   WARNING 0: Completion code = -2 trying new start** ** 

50 

51   WARNING 1: Completion code = -2 trying new start** ** 

52 

53   WARNING 2: Completion code = -2 trying new start** ** 

54 

55   WARNING 3: Completion code = -2 trying new start** ** 

56 

57   WARNING 4: Completion code = -2 trying new start** ** 

58 

59   WARNING 5: Completion code = -2 trying new start** ** 

60 
D-109 DRAFT – DO NOT CITE OR QUOTE 

1   WARNING 6: Completion code = -2 trying new start** ** 

2 

3   WARNING 7: Completion code = -2 trying new start** ** 

4 

5   WARNING 8: Completion code = -2 trying new start** ** 

6 

7   WARNING 9: Completion code = -2 trying new start** ** 

8 

9   WARNING: Completion code = -2. Optimum not found. Trying new starting 

10 point** ** 

11 

12   WARNING 0: Completion code = -2 trying new start** ** 

13 

14   WARNING 1: Completion code = -5 trying new start** ** 

15 

16   WARNING 2: Completion code = -2 trying new start** ** 

17 

18   WARNING 3: Completion code = -2 trying new start** ** 

19 

20   WARNING 4: Completion code = -2 trying new start** ** 

21 

22   WARNING 5: Completion code = -2 trying new start** ** 

23 

24   WARNING 6: Completion code = -2 trying new start** ** 

25 

26   WARNING 7: Completion code = -5 trying new start** ** 

27 

28   WARNING 8: Completion code = -2 trying new start** ** 

29 

30   WARNING 9: Completion code = -5 trying new start** ** 

31 

32 

33 Warning: completion code still negative 

34 BMDL did not converge for BMR = 0.000010 

35 

36 Program execution is stopped 
D-110 DRAFT – DO NOT CITE OR QUOTE 

1 RAVEH_HUB_MUT_BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\RAVEH_HUB_MUT_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\RAVEH_HUB_MUT_BAP.plt 

8 Wed Jun 29 12:15:41 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0.00102082 

41 **** WARNING: Completion code = -2. Optimum not found. Trying new starting 

42 pont  

43 

44 ** ** WARNING 0: Completion code = -2 trying new start  

45 

46 ** ** WARNING 1: Completion code = -2 trying new start  

47 

48 ** ** WARNING 2: Completion code = -2 trying new start  

49 

50 ** ** WARNING 3: Completion code = -2 trying new start  

51 

52 ** ** WARNING 4: Completion code = -2 trying new start  

53 

54 ** ** WARNING 5: Completion code = -2 trying new start  

55 

56 ** ** WARNING 6: Completion code = -2 trying new start  

57 

58 ** ** WARNING 7: Completion code = -2 trying new start  

59 

60 ** ** WARNING 8: Completion code = -2 trying new start  
D-111 DRAFT – DO NOT CITE OR QUOTE 

1 

2 ** ** WARNING 9: Completion code = -2 trying new start  

3 

4 ** ** WARNING: Completion code = -2. Optimum not found. Trying new starting 

5 point  

6 

7 ** ** WARNING 0: Completion code = -2 trying new start  

8 

9 ** ** WARNING 1: Completion code = -2 trying new start  

10 

11 ** ** WARNING 2: Completion code = -2 trying new start  

12 

13 ** ** WARNING 3: Completion code = -2 trying new start**** 

14 

15 

16 

17 Asymptotic Correlation Matrix of Parameter Estimates 

18 

19 Background Beta(1) 

20 

21 Background 1 -0.71 

22 

23 Beta(1) -0.71 1 

24 

25 

26 

27 Parameter Estimates 

28 

29 Variable Estimate Std. Err. 

30 Background 2.6399e-005 0.00257721 

31 Beta(1) 0.000947187 0.00419869 

32 

33 

34 

35 Analysis of Deviance Table 

36 

37 Model Log(likelihood) Deviance Test DF P-value 

38 Full model -1077.99 

39 Fitted model -1078.81 1.63811 1 0.2006 

40 Reduced model -1144.43 132.88 2 <.0001 

41 

42 AIC: 2161.62 

43 

44 

45 Goodness of Fit 

46 

47 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

48 ----------------------------------------------------------------------- 

49 i: 1 

50 0.0000 0.0000 2.640 3 100000 0.136 

51 i: 2 

52 0.3000 0.0003 31.051 25 100000 -0.195 

53 i: 3 

54 1.0000 0.0010 97.311 103 100000 0.059 

55 

56 Chi-square = 1.56 DF = 1 P-value = 0.2115 

57 

58 

59 Benchmark Dose Computation 

60 
D-112 DRAFT – DO NOT CITE OR QUOTE 

1 Specified effect = 0.0001 

2 

3 Risk Type = Extra risk 

4 

5 Confidence level = 0.95 

6 

7 BMD = 0.105581 

8 

9 BMDL = 0.0908465 
D-113 DRAFT – DO NOT CITE OR QUOTE 

1 RAVEH_HUB_MUT_cpcdp.OUT.txt 

2 ==================================================================== 

3 Quantal Linear Model $Revision: 2.2 $ $Date: 2000/03/17 22:27:16 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\RAVEH_HUB_MUT_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\RAVEH_HUB_MUT_BAP.plt 

8 Wed Jun 29 12:09:01 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP(-slope*dose)] 

17 

18 

19 Dependent variable = COLUMN2 

20 Independent variable = COLUMN1 

21 

22 Total number of observations = 3 

23 Total number of records with missing values = 0 

24 Maximum number of iterations = 250 

25 Relative Function Convergence has been set to: 1e-008 

26 Parameter Convergence has been set to: 1e-008 

27 

28 

29 

30 Default Initial (and Specified) Parameter Values 

31 Background = 3.49997e-005 

32 Slope = 0.000170019 

33 Power = 1 Specified 

34 

35 

36 Asymptotic Correlation Matrix of Parameter Estimates 

37 

38 ( *** The model parameter(s) -Power 

39 have been estimated at a boundary point, or have been 

40 specified by the user, 

41 and do not appear in the correlation matrix ) 

42 

43 Background Slope 

44 

45 Background 1 -0.51 

46 

47 Slope -0.51 1 

48 

49 

50 

51 Parameter Estimates 

52 

53 Variable Estimate Std. Err. 

54 Background 3.16959e-005 1.69176e-005 

55 Slope 0.000173022 4.78826e-005 

56 

57 

58 

59 Analysis of Deviance Table 

60 
D-114 DRAFT – DO NOT CITE OR QUOTE 

1 Model Log(likelihood) Deviance Test DF P-value 

2 Full model -317.426 

3 Fitted model -317.46 0.0679084 1 0.7944 

4 Reduced model -324.664 14.4766 2 0.0007185 

5 

6 AIC: 638.919 

7 

8 

9 Goodness of Fit 

10 

11 Scaled 

12 Dose Est._Prob. Expected Observed Size Residual 

13 ------------------------------------------------------------------------ 

14 0.0000 0.0000 3.170 3 100000 -0.09526 

15 0.3000 0.0001 8.360 9 100000 0.2214 

16 1.0000 0.0002 20.470 20 100000 -0.1038 

17 

18 Chi-square = 0.07 DF = 1 P-value = 0.7930 

19 

20 

21 Benchmark Dose Computation 

22 

23 Specified effect = 0.0001 

24 

25 Risk Type = Extra risk 

26 

27 Confidence level = 0.95 

28 

29 BMD = 0.577991 

30 

31 BMDL = 0.390507 

32 
D-115 DRAFT – DO NOT CITE OR QUOTE 

1 RAVEH_MUT_bap.OUT.txt 

2 ==================================================================== 

3 Quantal Linear Model $Revision: 2.2 $ $Date: 2000/03/17 22:27:16 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\RAVEH_MUT_CPCDP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\RAVEH_MUT_CPCDP.plt 

8 Wed Jun 29 12:33:35 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP(-slope*dose)] 

17 

18 

19 Dependent variable = COLUMN2 

20 Independent variable = COLUMN1 

21 

22 Total number of observations = 3 

23 Total number of records with missing values = 0 

24 Maximum number of iterations = 250 

25 Relative Function Convergence has been set to: 1e-008 

26 Parameter Convergence has been set to: 1e-008 

27 

28 

29 

30 Default Initial (and Specified) Parameter Values 

31 Background = 7.49999e-006 

32 Slope = 6.70027e-005 

33 Power = 1 Specified 

34 

35 

36 Asymptotic Correlation Matrix of Parameter Estimates 

37 

38 ( *** The model parameter(s) -Power 

39 have been estimated at a boundary point, or have been 

40 specified by the user, 

41 and do not appear in the correlation matrix ) 

42 

43 Background Slope 

44 

45 Background 1 -0.38 

46 

47 Slope -0.38 1 

48 

49 

50 

51 Parameter Estimates 

52 

53 Variable Estimate Std. Err. 

54 Background 6.11766e-006 2.23574e-006 

55 Slope 6.35766e-005 8.04156e-006 

56 

57 

58 

59 Analysis of Deviance Table 

60 
D-116 DRAFT – DO NOT CITE OR QUOTE 

1 Model Log(likelihood) Deviance Test DF P-value 

2 Full model -1104.33 

3 Fitted model -1105.09 1.53413 1 0.2155 

4 Reduced model -1141.2 73.7415 2 <.0001 

5 

6 AIC: 2214.19 

7 

8 

9 Goodness of Fit 

10 

11 Scaled 

12 Dose Est._Prob. Expected Observed Size Residual 

13 ------------------------------------------------------------------------ 

14 0.0000 0.0000 6.118 7 1000000 0.3567 

15 0.3000 0.0000 25.190 20 1000000 -1.034 

16 1.0000 0.0001 69.692 74 1000000 0.5161 

17 

18 Chi-square = 1.46 DF = 1 P-value = 0.2264 

19 

20 

21 Benchmark Dose Computation 

22 

23 Specified effect = 1e-005 

24 

25 Risk Type = Extra risk 

26 

27 Confidence level = 0.95 

28 

29 BMD = 0.157291 

30 

31 BMDL = 0.12931 

32 
D-117 DRAFT – DO NOT CITE OR QUOTE 

1 RAVEH_MUT_CPCDP.OUT.txt 

2 ==================================================================== 

3 Quantal Linear Model $Revision: 2.2 $ $Date: 2000/03/17 22:27:16 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\RAVEH_MUT_CPCDP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\RAVEH_MUT_CPCDP.plt 

8 Wed Jun 29 12:31:46 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP(-slope*dose)] 

17 

18 

19 Dependent variable = COLUMN2 

20 Independent variable = COLUMN1 

21 

22 Total number of observations = 4 

23 Total number of records with missing values = 0 

24 Maximum number of iterations = 250 

25 Relative Function Convergence has been set to: 1e-008 

26 Parameter Convergence has been set to: 1e-008 

27 

28 

29 

30 Default Initial (and Specified) Parameter Values 

31 Background = 1.5e-006 

32 Slope = 9.00013e-006 

33 Power = 1 Specified 

34 

35 

36 Asymptotic Correlation Matrix of Parameter Estimates 

37 

38 ( *** The model parameter(s) -Power 

39 have been estimated at a boundary point, or have been 

40 specified by the user, 

41 and do not appear in the correlation matrix ) 

42 

43 Background Slope 

44 

45 Background 1 -0.43 

46 

47 Slope -0.43 1 

48 

49 

50 

51 Parameter Estimates 

52 

53 Variable Estimate Std. Err. 

54 Background 1.26496e-006 1.07098e-006 

55 Slope 9.05599e-006 1.68076e-006 

56 

57 

58 

59 Analysis of Deviance Table 

60 
D-118 DRAFT – DO NOT CITE OR QUOTE 

1 Model Log(likelihood) Deviance Test DF P-value 

2 Full model -527.507 

3 Fitted model -527.666 0.317201 2 0.8533 

4 Reduced model -546.375 37.7352 3 <.0001 

5 

6 AIC: 1059.33 

7 

8 

9 Goodness of Fit 

10 

11 Scaled 

12 Dose Est._Prob. Expected Observed Size Residual 

13 ------------------------------------------------------------------------ 

14 0.0000 0.0000 1.265 1 1000000 -0.2356 

15 0.3000 0.0000 3.982 5 1000000 0.5103 

16 1.0000 0.0000 10.321 10 1000000 -0.09989 

17 3.0000 0.0000 28.433 28 1000000 -0.08112 

18 

19 Chi-square = 0.33 DF = 2 P-value = 0.8469 

20 

21 

22 Benchmark Dose Computation 

23 

24 Specified effect = 1e-005 

25 

26 Risk Type = Extra risk 

27 

28 Confidence level = 0.95 

29 

30 BMD = 1.10425 

31 

32 BMDL = 0.835597 

33 
D-119 DRAFT – DO NOT CITE OR QUOTE 

1 SLAGA_MUT_BAA.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\SLAGA_MUT_BAA.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\SLAGA_MUT_BAA.plt 

8 Thu Jul 07 15:25:30 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 7.29666e-005 

40 Beta(1) = 3.12233e-006 

41 **** WARNING: Completion code = 7. Optimum not found. Trying new starting 

42 pont  

43 

44 ** ** WARNING 0: Completion code = 7 trying new start  

45 

46 ** ** WARNING 1: Completion code = -2 trying new start  

47 

48 ** ** WARNING 2: Completion code = -2 trying new start  

49 

50 ** ** WARNING 3: Completion code = -2 trying new start  

51 

52 ** ** WARNING 4: Completion code = 7 trying new start  

53 

54 ** ** WARNING 5: Completion code = -2 trying new start  

55 

56 ** ** WARNING 6: Completion code = -2 trying new start  

57 

58 ** ** WARNING 7: Completion code = -2 trying new start  

59 

60 ** ** WARNING 8: Completion code = -2 trying new start  
D-120 DRAFT – DO NOT CITE OR QUOTE 

1 

2 ** ** WARNING 9: Completion code = 7 trying new start  

3 

4 ** ** WARNING: Completion code = -2. Optimum not found. Trying new starting 

5 point  

6 

7 ** ** WARNING 0: Completion code = -2 trying new start**** 

8 

9 

10 

11 Asymptotic Correlation Matrix of Parameter Estimates 

12 

13 Background Beta(1) 

14 

15 Background 1 -0.63 

16 

17 Beta(1) -0.63 1 

18 

19 

20 

21 Parameter Estimates 

22 

23 Variable Estimate Std. Err. 

24 Background 7.26607e-005 0.0023585 

25 Beta(1) 3.14129e-006 9.25599e-005 

26 

27 

28 

29 Analysis of Deviance Table 

30 

31 Model Log(likelihood) Deviance Test DF P-value 

32 Full model -365.644 

33 Fitted model -365.656 0.0243422 1 0.876 

34 Reduced model -370.021 8.75326 2 0.01257 

35 

36 AIC: 735.312 

37 

38 

39 Goodness of Fit 

40 

41 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

42 ----------------------------------------------------------------------- 

43 i: 1 

44 0.0000 0.0001 7.266 7 100000 -0.037 

45 i: 2 

46 4.4000 0.0001 8.648 9 100000 0.041 

47 i: 3 

48 44.0000 0.0002 21.086 21 100000 -0.004 

49 

50 Chi-square = 0.02 DF = 1 P-value = 0.8758 

51 

52 

53 Benchmark Dose Computation 

54 

55 Specified effect = 0.0001 

56 

57 Risk Type = Extra risk 

58 

59 Confidence level = 0.95 

60 
D-121 DRAFT – DO NOT CITE OR QUOTE 

1 BMD = 31.8356 

2 

3 BMDL = 19.0163 
D-122 DRAFT – DO NOT CITE OR QUOTE 

1 SLAGA_MUT_BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\SLAGA_MUT_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\SLAGA_MUT_BAP.plt 

8 Wed Jun 29 13:01:31 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.000214668 

40 Beta(1) = 0.00154564 

41 Beta(2) = 0.00022152 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) Beta(2) 

52 

53 Beta(1) 1 -0.98 

54 

55 Beta(2) -0.98 1 

56 

57 

58 

59 Parameter Estimates 

60 
D-123 DRAFT – DO NOT CITE OR QUOTE 

1 Variable Estimate Std. Err. 

2 Background 0 NA 

3 Beta(1) 0.00207246 0.0109511 

4 Beta(2) 9.74689e-005 0.00286413 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 Warning: Likelihood for the fitted model larger than the Likelihood for the 

11 full model. 

12 Error in computing chi-square; returning 2 

13 

14 

15 Analysis of Deviance Table 

16 

17 Model Log(likelihood) Deviance Test DF P-value 

18 Full model -823.498 

19 Fitted model -816.691 -13.6145 2 2 

20 Reduced model -907.084 167.172 3 <.0001 

21 

22 AIC: 1637.38 

23 

24 

25 Goodness of Fit 

26 

27 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

28 ----------------------------------------------------------------------- 

29 i: 1 

30 0.0000 0.0000 0.000 1 1000070000000.000 

31 i: 2 

32 0.4000 0.0008 8.442 11 10000 0.303 

33 i: 3 

34 1.3000 0.0029 28.548 25 10000 -0.125 

35 i: 4 

36 4.0000 0.0098 98.010 99 10000 0.010 

37 

38 Chi-square = 1.23 DF = 2 P-value = 0.5412 

39 

40 

41 Benchmark Dose Computation 

42 

43 Specified effect = 0.0001 

44 

45 Risk Type = Extra risk 

46 

47 Confidence level = 0.95 

48 

49 BMD = 0.0481451 

50 

51 BMDL = 0.0370516 

52 
D-124 DRAFT – DO NOT CITE OR QUOTE 

1 D.6. MALIGNANT TRANSFORMATION 

2 CASTO_MT_BAP.OUT.txt 

3 ==================================================================== 

4 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

5 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

6 RPS\MODELING\CASTO_MT_BAP.(d) 

7 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

8 DOCUMENTS\PAH RPS\MODELING\CASTO_MT_BAP.plt 

9 Thu Jun 23 13:30:59 2005 

10 ==================================================================== 

11 

12 BMDS MODEL RUN 

13 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

14 

15 The form of the probability function is: 

16 

17 P[response] = background + (1-background)*[1-EXP( 

18 -beta1*dose^1)] 

19 

20 The parameter betas are restricted to be positive 

21 

22 

23 Dependent variable = COLUMN2 

24 Independent variable = COLUMN1 

25 

26 Total number of observations = 3 

27 Total number of records with missing values = 0 

28 Total number of parameters in model = 2 

29 Total number of specified parameters = 0 

30 Degree of polynomial = 1 

31 

32 

33 Maximum number of iterations = 250 

34 Relative Function Convergence has been set to: 1e-008 

35 Parameter Convergence has been set to: 1e-008 

36 

37 

38 

39 Default Initial Parameter Values 

40 Background = 1.02144e-005 

41 Beta(1) = 7.98743e-005 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 
D-125 DRAFT – DO NOT CITE OR QUOTE 

1 Background 0 NA 

2 Beta(1) 9.62612e-005 0.00234809 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -185.57 

14 Fitted model -186.065 0.988828 2 0.6099 

15 Reduced model -192.98 14.82 2 0.0006052 

16 

17 AIC: 374.13 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 100000 0.000 

26 i: 2 

27 0.6200 0.0001 5.968 8 100000 0.340 

28 i: 3 

29 1.2500 0.0001 12.032 10 100000 -0.169 

30 

31 Chi-square = 1.04 DF = 2 P-value = 0.5960 

32 

33 

34 Benchmark Dose Computation 

35 

36 Specified effect = 1e-005 

37 

38 Risk Type = Extra risk 

39 

40 Confidence level = 0.95 

41 

42 BMD = 0.103885 

43 

44   WARNING: Completion code = -5. Optimum not found. Trying new starting 

45 point** ** 

46 

47   WARNING 0: Completion code = -5 trying new start** ** 

48 

49   WARNING 1: Completion code = -5 trying new start** ** 

50 

51   WARNING 2: Completion code = -5 trying new start** ** 

52 

53   WARNING 3: Completion code = -5 trying new start** ** 

54 

55   WARNING 4: Completion code = -5 trying new start** ** 

56 

57   WARNING 5: Completion code = -5 trying new start** ** 

58 

59   WARNING 6: Completion code = -5 trying new start** ** 

60 
D-126 DRAFT – DO NOT CITE OR QUOTE 

1   WARNING 7: Completion code = -5 trying new start** ** 

2 

3   WARNING 8: Completion code = -5 trying new start** ** 

4 

5   WARNING 9: Completion code = -5 trying new start** ** 

6 

7   WARNING: Completion code = -5. Optimum not found. Trying new starting 

8 point** ** 

9 

10 BMDL = 0.0721753 
D-127 DRAFT – DO NOT CITE OR QUOTE 

1 CASTO_MT_DBAHA.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\CASTO_MT_DBAHA.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\CASTO_MT_DBAHA.plt 

8 Thu Jun 23 13:32:00 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 3 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 2 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 1 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 6.92924e-008 

40 Beta(1) = 3.99789e-006 

41 **** WARNING: Completion code = -2. Optimum not found. Trying new starting 

42 pont  

43 

44 ** ** WARNING 0: Completion code = -2 trying new start  

45 

46 ** ** WARNING 1: Completion code = -2 trying new start  

47 

48 ** ** WARNING 2: Completion code = -2 trying new start  

49 

50 ** ** WARNING 3: Completion code = -2 trying new start  

51 

52 ** ** WARNING 4: Completion code = -2 trying new start  

53 

54 ** ** WARNING 5: Completion code = -2 trying new start  

55 

56 ** ** WARNING 6: Completion code = -2 trying new start  

57 

58 ** ** WARNING 7: Completion code = -2 trying new start  

59 

60 ** ** WARNING 8: Completion code = -2 trying new start  
D-128 DRAFT – DO NOT CITE OR QUOTE 

1 

2 ** ** WARNING 9: Completion code = -2 trying new start  

3 

4 ** ** WARNING: Completion code = -2. Optimum not found. Trying new starting 

5 point  

6 

7 

8 

9 Asymptotic Correlation Matrix of Parameter Estimates 

10 

11 ( ** * The model parameter(s) -Background 

12 have been estimated at a boundary point, or have been 

13 specified by the user, 

14 and do not appear in the correlation matrix ) 

15 

16 Beta(1) 

17 

18 Beta(1) 1 

19 

20 

21 

22 Parameter Estimates 

23 

24 Variable Estimate Std. Err. 

25 Background 0 NA 

26 Beta(1) 4.05407e-006 0.000361631 

27 

28 NA - Indicates that this parameter has hit a bound 

29 implied by some inequality constraint and thus

30 has no standard error. 

31 

32 

33 

34 Analysis of Deviance Table 

35 

36 Model Log(likelihood) Deviance Test DF P-value 

37 Full model -191.16 

38 Fitted model -191.162 0.00552866 2 0.9972 

39 Reduced model -198.091 13.863 2 0.0009765 

40 

41 AIC: 384.325 

42 

43 

44 Goodness of Fit 

45 

46 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

47 ----------------------------------------------------------------------- 

48 i: 1 

49 0.0000 0.0000 0.000 0 1000000 0.000 

50 i: 2 

51 1.2000 0.0000 4.865 5 1000000 0.028 

52 i: 3 

53 2.5000 0.0000 10.135 10 1000000 -0.013 

54 

55 Chi-square = 0.01 DF = 2 P-value = 0.9972 

56 

57 

58 Benchmark Dose Computation 

59 

60 Specified effect = 1e-005 
D-129 DRAFT – DO NOT CITE OR QUOTE 

1 

2 Risk Type = Extra risk 

3 

4 Confidence level = 0.95 

5 

6 BMD = 2.46667 

7 

8 **** WARNING: Completion code = -5. Optimum not found. Trying new starting 

9 point  

10 

11 ** ** WARNING 0: Completion code = -1 trying new start  

12 

13 ** ** WARNING 1: Completion code = -1 trying new start  

14 

15 ** ** WARNING 2: Completion code = -1 trying new start  

16 

17 ** ** WARNING 3: Completion code = -1 trying new start  

18 

19 ** ** WARNING 4: Completion code = -1 trying new start  

20 

21 ** ** WARNING 5: Completion code = -1 trying new start  

22 

23 ** ** WARNING 6: Completion code = -1 trying new start  

24 

25 ** ** WARNING 7: Completion code = -1 trying new start  

26 

27 ** ** WARNING 8: Completion code = -1 trying new start  

28 

29 ** ** WARNING 9: Completion code = -1 trying new start  

30 

31 ** ** WARNING: Completion code = -1. Optimum not found. Trying new starting 

32 point**** 

33 

34 BMDL = 1.65901 
D-130 DRAFT – DO NOT CITE OR QUOTE 

1 EMURA_MT_Baa.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\EMURA_MT_BBF.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\EMURA_MT_BBF.plt 

8 Thu Jun 23 15:46:49 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 6 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 5 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 4 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 6.24839e-005 

40 Beta(1) = 0.000973789 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 Beta(4) = 0 

44 

45 

46 Asymptotic Correlation Matrix of Parameter Estimates 

47 

48 ( *** The model parameter(s) -Background -Beta(2) -Beta(3) 

49 -Beta(4) 

50 have been estimated at a boundary point, or have been 

51 specified by the user, 

52 and do not appear in the correlation matrix ) 

53 

54 Beta(1) 

55 

56 Beta(1) 1 

57 

58 

59 

60 Parameter Estimates 
D-131 DRAFT – DO NOT CITE OR QUOTE 

1 

2 Variable Estimate Std. Err. 

3 Background 0 NA 

4 Beta(1) 0.00117377 0.0091424 

5 Beta(2) 0 NA 

6 Beta(3) 0 NA 

7 Beta(4) 0 NA 

8 

9 NA - Indicates that this parameter has hit a bound 

10 implied by some inequality constraint and thus

11 has no standard error. 

12 

13 

14 

15 Analysis of Deviance Table 

16 

17 Model Log(likelihood) Deviance Test DF P-value 

18 Full model -184.252 

19 Fitted model -185.671 2.83903 5 0.7248 

20 Reduced model -196.039 23.575 5 0.000262 

21 

22 AIC: 373.342 

23 

24 

25 Goodness of Fit 

26 

27 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

28 ----------------------------------------------------------------------- 

29 i: 1 

30 0.0000 0.0000 0.000 0 10000 0.000 

31 i: 2 

32 0.0250 0.0000 0.293 0 10000 -1.000 

33 i: 3 

34 0.1000 0.0001 1.174 3 10000 1.556 

35 i: 4 

36 0.2500 0.0003 2.934 3 10000 0.023 

37 i: 5 

38 0.5000 0.0006 5.867 6 10000 0.023 

39 i: 6 

40 1.0000 0.0012 11.731 10 10000 -0.148 

41 

42 Chi-square = 3.40 DF = 5 P-value = 0.6392 

43 

44 

45 Benchmark Dose Computation 

46 

47 Specified effect = 0.001 

48 

49 Risk Type = Extra risk 

50 

51 Confidence level = 0.95 

52 

53 BMD = 0.85238 

54 BMDL = 0.611981 

55 EMURA_MT_BBF.OUT.txt 

56 ==================================================================== 

57 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

58 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

59 RPS\MODELING\EMURA_MT_BBF.(d) 
D-132 DRAFT – DO NOT CITE OR QUOTE 

1 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

2 DOCUMENTS\PAH RPS\MODELING\EMURA_MT_BBF.plt 

3 Thu Jun 23 15:37:20 2005 

4 ==================================================================== 

5 

6 BMDS MODEL RUN 

7 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

8 

9 The form of the probability function is: 

10 

11 P[response] = background + (1-background)*[1-EXP( 

12 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

13 

14 The parameter betas are restricted to be positive 

15 

16 

17 Dependent variable = COLUMN2 

18 Independent variable = COLUMN1 

19 

20 Total number of observations = 6 

21 Total number of records with missing values = 0 

22 Total number of parameters in model = 5 

23 Total number of specified parameters = 0 

24 Degree of polynomial = 4 

25 

26 

27 Maximum number of iterations = 250 

28 Relative Function Convergence has been set to: 1e-008 

29 Parameter Convergence has been set to: 1e-008 

30 

31 

32 

33 Default Initial Parameter Values 

34 Background = 6.48647e-005 

35 Beta(1) = 0.00111706 

36 Beta(2) = 0 

37 Beta(3) = 1.51794e-005 

38 Beta(4) = 0 

39 

40 

41 Asymptotic Correlation Matrix of Parameter Estimates 

42 

43 ( *** The model parameter(s) -Background -Beta(2) -Beta(3) 

44 -Beta(4) 

45 have been estimated at a boundary point, or have been 

46 specified by the user, 

47 and do not appear in the correlation matrix ) 

48 

49 Beta(1) 

50 

51 Beta(1) 1 

52 

53 

54 

55 Parameter Estimates 

56 

57 Variable Estimate Std. Err. 

58 Background 0 NA 

59 Beta(1) 0.00133391 0.00909075 

60 Beta(2) 0 NA 
D-133 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(3) 0 NA 

2 Beta(4) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -205.838 

14 Fitted model -208.019 4.36272 5 0.4985 

15 Reduced model -219.575 27.4752 5 <.0001 

16 

17 AIC: 418.038 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 10000 0.000 

26 i: 2 

27 0.0250 0.0000 0.333 0 10000 -1.000 

28 i: 3 

29 0.1000 0.0001 1.334 4 10000 1.999 

30 i: 4 

31 0.2500 0.0003 3.334 3 10000 -0.100 

32 i: 5 

33 0.5000 0.0007 6.667 6 10000 -0.100 

34 i: 6 

35 1.0000 0.0013 13.330 12 10000 -0.100 

36 

37 Chi-square = 5.90 DF = 5 P-value = 0.3164 

38 

39 

40 Benchmark Dose Computation 

41 

42 Specified effect = 0.001 

43 

44 Risk Type = Extra risk 

45 

46 Confidence level = 0.95 

47 

48 BMD = 0.750052 

49 BMDL = 0.54909 
D-134 DRAFT – DO NOT CITE OR QUOTE 

1 EMURA_MT_I_BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\EMURA_MT_I_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\EMURA_MT_I_BAP.plt 

8 Thu Jun 23 15:28:17 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 5 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 4 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 3 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 6.51885e-005 

40 Beta(1) = 0.021934 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 

44 

45 Asymptotic Correlation Matrix of Parameter Estimates 

46 

47 ( *** The model parameter(s) -Background -Beta(2) -Beta(3) 

48 have been estimated at a boundary point, or have been 

49 specified by the user, 

50 and do not appear in the correlation matrix ) 

51 

52 Beta(1) 

53 

54 Beta(1) 1 

55 

56 

57 

58 Parameter Estimates 

59 

60 Variable Estimate Std. Err. 
D-135 DRAFT – DO NOT CITE OR QUOTE 

1 Background 0 NA 

2 Beta(1) 0.0227293 0.0369378 

3 Beta(2) 0 NA 

4 Beta(3) 0 NA 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -614.919 

16 Fitted model -618.123 6.40862 4 0.1706 

17 Reduced model -677.621 125.404 4 <.0001 

18 

19 AIC: 1238.25 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0000 0.000 0 10000 0.000 

28 i: 2 

29 0.0100 0.0002 2.273 0 10000 -1.000 

30 i: 3 

31 0.0500 0.0011 11.358 11 10000 -0.032 

32 i: 4 

33 0.1000 0.0023 22.703 29 10000 0.278 

34 i: 5 

35 0.2500 0.0057 56.662 53 10000 -0.065 

36 

37 Chi-square = 4.27 DF = 4 P-value = 0.3703 

38 

39 

40 Benchmark Dose Computation 

41 

42 Specified effect = 0.001 

43 

44 Risk Type = Extra risk 

45 

46 Confidence level = 0.95 

47 

48 BMD = 0.0440182 

49 

50 BMDL = 0.037291 
D-136 DRAFT – DO NOT CITE OR QUOTE 

1 EMURA_MT_II_BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\EMURA_MT_II_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\EMURA_MT_II_BAP.plt 

8 Thu Jun 23 15:54:16 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 5 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 4 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 3 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0002687 

40 Beta(1) = 0.0184676 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 

44 

45 Asymptotic Correlation Matrix of Parameter Estimates 

46 

47 ( *** The model parameter(s) -Background -Beta(2) -Beta(3) 

48 have been estimated at a boundary point, or have been 

49 specified by the user, 

50 and do not appear in the correlation matrix ) 

51 

52 Beta(1) 

53 

54 Beta(1) 1 

55 

56 

57 

58 Parameter Estimates 

59 

60 Variable Estimate Std. Err. 
D-137 DRAFT – DO NOT CITE OR QUOTE 

1 Background 0 NA 

2 Beta(1) 0.021747 0.0381969 

3 Beta(2) 0 NA 

4 Beta(3) 0 NA 

5 

6 NA - Indicates that this parameter has hit a bound 

7 implied by some inequality constraint and thus

8 has no standard error. 

9 

10 

11 

12 Analysis of Deviance Table 

13 

14 Model Log(likelihood) Deviance Test DF P-value 

15 Full model -606.226 

16 Fitted model -608.64 4.82649 4 0.3056 

17 Reduced model -652.392 92.3321 4 <.0001 

18 

19 AIC: 1219.28 

20 

21 

22 Goodness of Fit 

23 

24 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

25 ----------------------------------------------------------------------- 

26 i: 1 

27 0.0000 0.0000 0.000 0 10000 0.000 

28 i: 2 

29 0.0100 0.0002 2.174 4 10000 0.840 

30 i: 3 

31 0.0500 0.0011 10.868 10 10000 -0.080 

32 i: 4 

33 0.1000 0.0022 21.723 29 10000 0.336 

34 i: 5 

35 0.2500 0.0054 54.220 46 10000 -0.152 

36 

37 Chi-square = 5.30 DF = 4 P-value = 0.2581 

38 

39 

40 Benchmark Dose Computation 

41 

42 Specified effect = 0.001 

43 

44 Risk Type = Extra risk 

45 

46 Confidence level = 0.95 

47 

48 BMD = 0.0460064 

49 

50 BMDL = 0.0388361 
D-138 DRAFT – DO NOT CITE OR QUOTE 

1 EMURA_MT_IP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\EMURA_MT_IP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\EMURA_MT_IP.plt 

8 Thu Jun 23 15:50:44 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 6 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 5 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 4 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 7.12074e-005 

40 Beta(1) = 0.00099924 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 Beta(4) = 0 

44 

45 

46 Asymptotic Correlation Matrix of Parameter Estimates 

47 

48 ( *** The model parameter(s) -Background -Beta(2) -Beta(3) 

49 -Beta(4) 

50 have been estimated at a boundary point, or have been 

51 specified by the user, 

52 and do not appear in the correlation matrix ) 

53 

54 Beta(1) 

55 

56 Beta(1) 1 

57 

58 

59 

60 Parameter Estimates 
D-139 DRAFT – DO NOT CITE OR QUOTE 

1 

2 Variable Estimate Std. Err. 

3 Background 0 NA 

4 Beta(1) 0.00122714 0.00918598 

5 Beta(2) 0 NA 

6 Beta(3) 0 NA 

7 Beta(4) 0 NA 

8 

9 NA - Indicates that this parameter has hit a bound 

10 implied by some inequality constraint and thus

11 has no standard error. 

12 

13 

14 

15 Analysis of Deviance Table 

16 

17 Model Log(likelihood) Deviance Test DF P-value 

18 Full model -191.591 

19 Fitted model -193.089 2.99724 5 0.7004 

20 Reduced model -203.928 24.6739 5 0.0001611 

21 

22 AIC: 388.178 

23 

24 

25 Goodness of Fit 

26 

27 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

28 ----------------------------------------------------------------------- 

29 i: 1 

30 0.0000 0.0000 0.000 0 10000 0.000 

31 i: 2 

32 0.0250 0.0000 0.307 0 10000 -1.000 

33 i: 3 

34 0.1000 0.0001 1.227 3 10000 1.445 

35 i: 4 

36 0.2500 0.0003 3.067 3 10000 -0.022 

37 i: 5 

38 0.5000 0.0006 6.134 7 10000 0.141 

39 i: 6 

40 1.0000 0.0012 12.264 10 10000 -0.185 

41 

42 Chi-square = 3.41 DF = 5 P-value = 0.6369 

43 

44 

45 Benchmark Dose Computation 

46 

47 Specified effect = 0.001 

48 

49 Risk Type = Extra risk 

50 

51 Confidence level = 0.95 

52 

53 BMD = 0.815309 

54 BMDL = 0.589412 
D-140 DRAFT – DO NOT CITE OR QUOTE 

1 LUBET_MT_BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\LUBET_MT_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\LUBET_MT_BAP.plt 

8 Thu Jun 23 16:11:06 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0617408 

40 Beta(1) = 0.0378355 

41 Beta(2) = 0 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(2) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(1) 

52 

53 Beta(1) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-141 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 0.056828 0.0340172 

2 Beta(2) 0 NA 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -21.9204 

14 Fitted model -22.8416 1.84243 3 0.6057 

15 Reduced model -27.0337 10.2266 3 0.01674 

16 

17 AIC: 47.6832 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 15 0.000 

26 i: 2 

27 1.0000 0.0552 0.829 1 15 0.219 

28 i: 3 

29 3.0000 0.1567 2.351 4 15 0.832 

30 i: 4 

31 10.0000 0.4335 6.503 5 15 -0.408 

32 

33 Chi-square = 2.02 DF = 3 P-value = 0.5679 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 0.1 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 1.85403 

45 

46 BMDL = 1.14367 
D-142 DRAFT – DO NOT CITE OR QUOTE 

1 LUBET_MT_BeP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\LUBET_MT_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\LUBET_MT_BAP.plt 

8 Thu Jun 23 16:14:09 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 4 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 3 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 2 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0.000632445 

41 Beta(2) = 5.70088e-005 

42 

43 

44 Asymptotic Correlation Matrix of Parameter Estimates 

45 

46 ( *** The model parameter(s) -Background -Beta(1) 

47 have been estimated at a boundary point, or have been 

48 specified by the user, 

49 and do not appear in the correlation matrix ) 

50 

51 Beta(2) 

52 

53 Beta(2) 1 

54 

55 

56 

57 Parameter Estimates 

58 

59 Variable Estimate Std. Err. 

60 Background 0 NA 
D-143 DRAFT – DO NOT CITE OR QUOTE 

1 Beta(1) 0 NA 

2 Beta(2) 6.35618e-005 3.53139e-005 

3 

4 NA - Indicates that this parameter has hit a bound 

5 implied by some inequality constraint and thus

6 has no standard error. 

7 

8 

9 

10 Analysis of Deviance Table 

11 

12 Model Log(likelihood) Deviance Test DF P-value 

13 Full model -14.0378 

14 Fitted model -14.1501 0.224517 3 0.9735 

15 Reduced model -23.5605 19.0453 3 0.0002676 

16 

17 AIC: 30.3001 

18 

19 

20 Goodness of Fit 

21 

22 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

23 ----------------------------------------------------------------------- 

24 i: 1 

25 0.0000 0.0000 0.000 0 15 0.000 

26 i: 2 

27 10.0000 0.0063 0.095 0 15 -1.006 

28 i: 3 

29 30.0000 0.0556 0.834 1 15 0.211 

30 i: 4 

31 100.0000 0.4704 7.056 7 15 -0.015 

32 

33 Chi-square = 0.13 DF = 3 P-value = 0.9878 

34 

35 

36 Benchmark Dose Computation 

37 

38 Specified effect = 0.1 

39 

40 Risk Type = Extra risk 

41 

42 Confidence level = 0.95 

43 

44 BMD = 40.7137 

45 

46 BMDL = 18.2541 
D-144 DRAFT – DO NOT CITE OR QUOTE 

1 MOHAPATRA_MT_BJAC.txt

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\_PAH 

5 RPS\MODELING\MOHAPATRA_MT_BJAC.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\_PAH RPS\MODELING\MOHAPATRA_MT_BJAC.plt 

8 Thu Feb 08 10:11:06 2007 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 6 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 5 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 4 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0 

40 Beta(1) = 0 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 Beta(4) = 6.31048e+018 

44 

45 

46 Asymptotic Correlation Matrix of Parameter Estimates 

47 

48 ( *** The model parameter(s) -Background -Beta(2) -Beta(3) 

49 have been estimated at a boundary point, or have been 

50 specified by the user, 

51 and do not appear in the correlation matrix ) 

52 

53 Beta(1) Beta(4) 

54 

55 Beta(1) 1 -0.73 

56 

57 Beta(4) -0.73 1 

58 

59 

60 
D-145 DRAFT – DO NOT CITE OR QUOTE 

1 Parameter Estimates 

2 

3 Variable Estimate Std. Err. 

4 Background 0 NA 

5 Beta(1) 2.44509 0.568863 

6 Beta(2) 0 NA 

7 Beta(3) 0 NA 

8 Beta(4) 0.332129 0.778407 

9 

10 NA - Indicates that this parameter has hit a bound 

11 implied by some inequality constraint and thus

12 has no standard error. 

13 

14 

15 

16 Analysis of Deviance Table 

17 

18 Model Log(likelihood) Deviance Test DF P-value 

19 Full model -64.5493 

20 Fitted model -64.8387 0.578751 4 0.9654 

21 Reduced model -198.931 268.764 5 <.0001 

22 

23 AIC: 133.677 

24 

25 

26 Goodness of Fit 

27 

28 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

29 ----------------------------------------------------------------------- 

30 i: 1 

31 0.0000 0.0000 0.000 0 48 0.000 

32 i: 2 

33 0.0100 0.0242 1.159 2 48 0.743 

34 i: 3 

35 0.0500 0.1151 5.524 5 48 -0.107 

36 i: 4 

37 0.5000 0.7116 34.155 34 48 -0.016 

38 i: 5 

39 1.0000 0.9378 45.014 45 48 -0.005 

40 i: 6 

41 2.0000 1.0000 47.998 48 48 1.000 

42 

43 Chi-square = 0.68 DF = 4 P-value = 0.9532 

44 

45 

46 Benchmark Dose Computation 

47 

48 Specified effect = 0.92 

49 

50 Risk Type = Extra risk 

51 

52 Confidence level = 0.95 

53 

54 BMD = 0.930952 

55 

56 BMDL = 0.766826 
D-146 DRAFT – DO NOT CITE OR QUOTE 

1 MOHAPATRA_MT_BLAC.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\_PAH 

5 RPS\MODELING\MOHAPATRA_MT_BLAC.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\_PAH RPS\MODELING\MOHAPATRA_MT_BLAC.plt 

8 Thu Feb 08 10:13:14 2007 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 6 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 5 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 4 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0997842 

40 Beta(1) = 0.189801 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 Beta(4) = 0 

44 

45 

46 Asymptotic Correlation Matrix of Parameter Estimates 

47 

48 ( *** The model parameter(s) -Background -Beta(2) -Beta(3) 

49 -Beta(4) 

50 have been estimated at a boundary point, or have been 

51 specified by the user, 

52 and do not appear in the correlation matrix ) 

53 

54 Beta(1) 

55 

56 Beta(1) 1 

57 

58 

59 

60 Parameter Estimates 
D-147 DRAFT – DO NOT CITE OR QUOTE 

1 

2 Variable Estimate Std. Err. 

3 Background 0 NA 

4 Beta(1) 0.237265 0.0278061 

5 Beta(2) 0 NA 

6 Beta(3) 0 NA 

7 Beta(4) 0 NA 

8 

9 NA - Indicates that this parameter has hit a bound 

10 implied by some inequality constraint and thus

11 has no standard error. 

12 

13 

14 

15 Analysis of Deviance Table 

16 

17 Model Log(likelihood) Deviance Test DF P-value 

18 Full model -159.727 

19 Fitted model -161.509 3.56545 5 0.6135 

20 Reduced model -243.072 166.691 5 <.0001 

21 

22 AIC: 325.019 

23 

24 

25 Goodness of Fit 

26 

27 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

28 ----------------------------------------------------------------------- 

29 i: 1 

30 0.0000 0.0000 0.000 0 60 0.000 

31 i: 2 

32 0.5000 0.1119 6.712 8 60 0.216 

33 i: 3 

34 1.0000 0.2112 12.673 14 60 0.133 

35 i: 4 

36 2.5000 0.4474 26.845 31 60 0.280 

37 i: 5 

38 5.0000 0.6947 41.679 42 60 0.025 

39 i: 6 

40 10.0000 0.9068 54.406 51 60 -0.671 

41 

42 Chi-square = 3.91 DF = 5 P-value = 0.5620 

43 

44 

45 Benchmark Dose Computation 

46 

47 Specified effect = 0.83 

48 

49 Risk Type = Extra risk 

50 

51 Confidence level = 0.95 

52 

53 BMD = 7.46828 

54 

55 BMDL = 6.45083 
D-148 DRAFT – DO NOT CITE OR QUOTE 

1 MOHAPATRA_MT_BEAC.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\_PAH 

5 RPS\MODELING\MOHAPATRA_MT_BEAC.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\_PAH RPS\MODELING\MOHAPATRA_MT_BEAC.plt 

8 Fri Feb 09 10:49:12 2007 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 6 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 5 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 4 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.0946116 

40 Beta(1) = 0.082434 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 Beta(4) = 0 

44 

45 

46 Asymptotic Correlation Matrix of Parameter Estimates 

47 

48 ( *** The model parameter(s) -Beta(2) -Beta(3) -Beta(4) 

49 have been estimated at a boundary point, or have been 

50 specified by the user, 

51 and do not appear in the correlation matrix ) 

52 

53 Background Beta(1) 

54 

55 Background 1 -0.68 

56 

57 Beta(1) -0.68 1 

58 

59 

60 
D-149 DRAFT – DO NOT CITE OR QUOTE 

1 Parameter Estimates 

2 

3 Variable Estimate Std. Err. 

4 Background 0.0246825 0.106613 

5 Beta(1) 0.109348 0.0321778 

6 Beta(2) 0 NA 

7 Beta(3) 0 NA 

8 Beta(4) 0 NA 

9 

10 NA - Indicates that this parameter has hit a bound 

11 implied by some inequality constraint and thus

12 has no standard error. 

13 

14 

15 

16 Analysis of Deviance Table 

17 

18 Model Log(likelihood) Deviance Test DF P-value 

19 Full model -101.226 

20 Fitted model -104.24 6.02698 4 0.1971 

21 Reduced model -126.655 50.8576 5 <.0001 

22 

23 AIC: 212.479 

24 

25 

26 Goodness of Fit 

27 

28 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

29 ----------------------------------------------------------------------- 

30 i: 1 

31 0.0000 0.0247 0.889 0 36 -1.025 

32 i: 2 

33 0.5000 0.0766 2.757 4 36 0.488 

34 i: 3 

35 1.0000 0.1257 4.525 6 36 0.373 

36 i: 4 

37 2.5000 0.2580 9.287 13 36 0.539 

38 i: 5 

39 5.0000 0.4355 15.676 15 36 -0.076 

40 i: 6 

41 10.0000 0.6732 24.236 21 36 -0.409 

42 

43 Chi-square = 5.44 DF = 4 P-value = 0.2448 

44 

45 

46 Benchmark Dose Computation 

47 

48 Specified effect = 0.86 

49 

50 Risk Type = Extra risk 

51 

52 Confidence level = 0.95 

53 

54 BMD = 17.9803 

55 

56 BMDL = 12.7064 
D-150 DRAFT – DO NOT CITE OR QUOTE 

1 PIENTA_MT_BAA.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\PIENTA_MT_BAA.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\PIENTA_MT_BAA.plt 

8 Tue Jul 05 13:52:46 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 6 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 5 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 4 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.00472474 

40 Beta(1) = 0 

41 Beta(2) = 0 

42 Beta(3) = 2.31177e-005 

43 Beta(4) = 0 

44 

45 

46 Asymptotic Correlation Matrix of Parameter Estimates 

47 

48 ( *** The model parameter(s) -Beta(1) -Beta(2) -Beta(3) 

49 have been estimated at a boundary point, or have been 

50 specified by the user, 

51 and do not appear in the correlation matrix ) 

52 

53 Background Beta(4) 

54 

55 Background 1 -0.43 

56 

57 Beta(4) -0.43 1 

58 

59 

60 
D-151 DRAFT – DO NOT CITE OR QUOTE 

1 Parameter Estimates 

2 

3 Variable Estimate Std. Err. 

4 Background 0.00480466 0.0290234 

5 Beta(1) 0 NA 

6 Beta(2) 0 NA 

7 Beta(3) 0 NA 

8 Beta(4) 2.25394e-006 6.9765e-006 

9 

10 NA - Indicates that this parameter has hit a bound 

11 implied by some inequality constraint and thus

12 has no standard error. 

13 

14 

15 

16 Analysis of Deviance Table 

17 

18 Model Log(likelihood) Deviance Test DF P-value 

19 Full model -67.8785 

20 Fitted model -69.9491 4.14115 4 0.3872 

21 Reduced model -74.327 12.8971 5 0.02436 

22 

23 AIC: 143.898 

24 

25 

26 Goodness of Fit 

27 

28 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

29 ----------------------------------------------------------------------- 

30 i: 1 

31 0.0000 0.0048 1.100 0 229 -1.005 

32 i: 2 

33 0.1000 0.0048 1.081 1 225 -0.075 

34 i: 3 

35 0.5000 0.0048 1.211 2 252 0.655 

36 i: 4 

37 1.0000 0.0048 0.928 2 193 1.161 

38 i: 5 

39 5.0000 0.0062 1.936 1 312 -0.487 

40 i: 6 

41 10.0000 0.0270 6.746 7 250 0.039 

42 

43 Chi-square = 3.34 DF = 4 P-value = 0.5028 

44 

45 

46 Benchmark Dose Computation 

47 

48 Specified effect = 0.01 

49 

50 Risk Type = Extra risk 

51 

52 Confidence level = 0.95 

53 

54 BMD = 8.17165 

55 

56   WARNING: Completion code = -2. Optimum not found. Trying new starting 

57 point** ** 

58 

59 BMDL = 4.47767 
D-152 DRAFT – DO NOT CITE OR QUOTE 

1 PIENTA_MT_BAP.OUT.txt 

2 ==================================================================== 

3 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

4 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

5 RPS\MODELING\PIENTA_MT_BAP.(d) 

6 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

7 DOCUMENTS\PAH RPS\MODELING\PIENTA_MT_BAP.plt 

8 Mon Jun 27 16:28:28 2005 

9 ==================================================================== 

10 

11 BMDS MODEL RUN 

12 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

13 

14 The form of the probability function is: 

15 

16 P[response] = background + (1-background)*[1-EXP( 

17 -beta1*dose^1-beta2*dose^2-beta3*dose^3-beta4*dose^4)] 

18 

19 The parameter betas are restricted to be positive 

20 

21 

22 Dependent variable = COLUMN2 

23 Independent variable = COLUMN1 

24 

25 Total number of observations = 5 

26 Total number of records with missing values = 0 

27 Total number of parameters in model = 5 

28 Total number of specified parameters = 0 

29 Degree of polynomial = 4 

30 

31 

32 Maximum number of iterations = 250 

33 Relative Function Convergence has been set to: 1e-008 

34 Parameter Convergence has been set to: 1e-008 

35 

36 

37 

38 Default Initial Parameter Values 

39 Background = 0.00129459 

40 Beta(1) = 0.00056154 

41 Beta(2) = 0 

42 Beta(3) = 0 

43 Beta(4) = 0 

44 

45 

46 Asymptotic Correlation Matrix of Parameter Estimates 

47 

48 ( *** The model parameter(s) -Beta(2) -Beta(3) -Beta(4) 

49 have been estimated at a boundary point, or have been 

50 specified by the user, 

51 and do not appear in the correlation matrix ) 

52 

53 Background Beta(1) 

54 

55 Background 1 -0.72 

56 

57 Beta(1) -0.72 1 

58 

59 

60 
D-153 DRAFT – DO NOT CITE OR QUOTE 

1 Parameter Estimates 

2 

3 Variable Estimate Std. Err. 

4 Background 0.000529694 0.0310484 

5 Beta(1) 0.000662444 0.00321227 

6 Beta(2) 0 NA 

7 Beta(3) 0 NA 

8 Beta(4) 0 NA 

9 

10 NA - Indicates that this parameter has hit a bound 

11 implied by some inequality constraint and thus

12 has no standard error. 

13 

14 

15 

16 Analysis of Deviance Table 

17 

18 Model Log(likelihood) Deviance Test DF P-value 

19 Full model -64.5099 

20 Fitted model -65.0987 1.17762 3 0.7584 

21 Reduced model -68.985 8.95024 4 0.06236 

22 

23 AIC: 134.197 

24 

25 

26 Goodness of Fit 

27 

28 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

29 ----------------------------------------------------------------------- 

30 i: 1 

31 0.0000 0.0005 0.267 0 504 -1.001 

32 i: 2 

33 1.0000 0.0012 0.468 1 393 1.137 

34 i: 3 

35 5.0000 0.0038 1.557 2 406 0.286 

36 i: 4 

37 10.0000 0.0071 3.094 3 434 -0.031 

38 i: 5 

39 20.0000 0.0137 5.611 5 410 -0.110 

40 

41 Chi-square = 1.07 DF = 3 P-value = 0.7847 

42 

43 

44 Benchmark Dose Computation 

45 

46 Specified effect = 0.01 

47 

48 Risk Type = Extra risk 

49 

50 Confidence level = 0.95 

51 

52 BMD = 15.1716 

53 

54 BMDL = 8.76437 

55 PIENTA_MT_DBAHA.OUT.txt 

56 ==================================================================== 

57 Multistage Model. $Revision: 2.1 $ $Date: 2000/08/21 03:38:21 $ 

58 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

59 RPS\MODELING\PIENTA_MT_DBAHA.(d) 
D-154 DRAFT – DO NOT CITE OR QUOTE 

1 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

2 DOCUMENTS\PAH RPS\MODELING\PIENTA_MT_DBAHA.plt 

3 Mon Jun 27 16:35:08 2005 

4 ==================================================================== 

5 

6 BMDS MODEL RUN 

7 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

8 

9 The form of the probability function is: 

10 

11 P[response] = background + (1-background)*[1-EXP( 

12 -beta1*dose^1-beta2*dose^2)] 

13 

14 The parameter betas are restricted to be positive 

15 

16 

17 Dependent variable = COLUMN2 

18 Independent variable = COLUMN1 

19 

20 Total number of observations = 4 

21 Total number of records with missing values = 0 

22 Total number of parameters in model = 3 

23 Total number of specified parameters = 0 

24 Degree of polynomial = 2 

25 

26 

27 Maximum number of iterations = 250 

28 Relative Function Convergence has been set to: 1e-008 

29 Parameter Convergence has been set to: 1e-008 

30 

31 

32 

33 Default Initial Parameter Values 

34 Background = 0.000660992 

35 Beta(1) = 0.020798 

36 Beta(2) = 0 

37 

38 

39 Asymptotic Correlation Matrix of Parameter Estimates 

40 

41 ( *** The model parameter(s) -Background -Beta(2) 

42 have been estimated at a boundary point, or have been 

43 specified by the user, 

44 and do not appear in the correlation matrix ) 

45 

46 Beta(1) 

47 

48 Beta(1) 1 

49 

50 

51 

52 Parameter Estimates 

53 

54 Variable Estimate Std. Err. 

55 Background 0 NA 

56 Beta(1) 0.0227021 0.0618036 

57 Beta(2) 0 NA 

58 

59 NA - Indicates that this parameter has hit a bound 

60 implied by some inequality constraint and thus
D-155 DRAFT – DO NOT CITE OR QUOTE 

1 has no standard error. 

2 

3 

4 

5 Analysis of Deviance Table 

6 

7 Model Log(likelihood) Deviance Test DF P-value 

8 Full model -40.1618 

9 Fitted model -41.0551 1.78665 3 0.6178 

10 Reduced model -45.7301 11.1367 3 0.01101 

11 

12 AIC: 84.1102 

13 

14 

15 Goodness of Fit 

16 

17 Dose Est._Prob. Expected Observed Size Chi^2 Res. 

18 ----------------------------------------------------------------------- 

19 i: 1 

20 0.0000 0.0000 0.000 0 229 0.000 

21 i: 2 

22 0.1000 0.0023 0.497 0 219 -1.002 

23 i: 3 

24 0.5000 0.0113 2.630 4 233 0.527 

25 i: 4 

26 1.0000 0.0224 4.871 4 217 -0.183 

27 

28 Chi-square = 1.38 DF = 3 P-value = 0.7105 

29 

30 

31 Benchmark Dose Computation 

32 

33 Specified effect = 0.01 

34 

35 Risk Type = Extra risk 

36 

37 Confidence level = 0.95 

38 

39 BMD = 0.442705 

40 

41 BMDL = 0.260515 

42 

43 
D-156 DRAFT – DO NOT CITE OR QUOTE 

1 D.7. IN VITRO DNA DAMAGE 

2 JOHNSEN_DNA_DAM_BJAC.OUT.txt 

3 ==================================================================== 

4 Polynomial Model. Revision: 2.2 Date: 9/12/2002 

5 Input Data File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY DOCUMENTS\PAH 

6 RPS\MODELING\JOHNSEN_DNA_DAM_BAP.(d) 

7 Gnuplot Plotting File: C:\DOCUMENTS AND SETTINGS\HCLYNCH\MY 

8 DOCUMENTS\PAH RPS\MODELING\JOHNSEN_DNA_DAM_BAP.plt 

9 Mon Jul 04 21:51:27 2005 

10 ==================================================================== 

11 

12 BMDS MODEL RUN 

13 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 

14 

15 The form of the response function is: 

16 

17 Y[dose] = beta_0 + beta_1*dose + beta_2*dose^2 + ... 

18 

19 

20 Dependent variable = MEAN 

21 Independent variable = COLUMN1 

22 rho is set to 0 

23 Signs of the polynomial coefficients are not restricted 

24 A constant variance model is fit 

25 

26 Total number of dose groups = 3 

27 Total number of records with missing values = 0 

28 Maximum number of iterations = 250 

29 Relative Function Convergence has been set to: 1e-008 

30 Parameter Convergence has been set to: 1e-008 

31 

32 

33 

34 Default Initial Parameter Values 

35 alpha = 5.88667 

36 rho = 0 Specified 

37 beta_0 = 4.94396 

38 beta_1 = 0.150549 

39 

40 

41 

42 Parameter Estimates 

43 

44 95.0% Wald 

45 Confidence Interval 

46 Variable Estimate Std. Err. Lower Conf. Limit 

47 Upper Conf. Limit 

48 alpha 4.14606 1.95447 0.315366 

49 7.97675 

50 beta_0 4.94396 0.875754 3.22751 

51 6.6604 

52 beta_1 0.150549 0.0503107 0.0519422 

53 0.249157 

54 

55 

56 Asymptotic Correlation Matrix of Parameter Estimates 

57 

58 alpha beta_0 beta_1 

59 alpha 1 7.6e-015 1.7e-015 
D-157 DRAFT – DO NOT CITE OR QUOTE 

1 beta_0 7.6e-015 1 -0.63 

2 beta_1 1.7e-015 -0.63 1 

3 

4 

5 Table of Data and Estimated Values of Interest

6 

7 Dose N Obs Mean Obs Std Dev Est Mean Est Std Dev Chi^2 

8 Res. 

9 ------ --- -------- ----------- -------- ----------- ---------

10 - 

11 

12 0 3 4.4 1.3 4.94 2.04 -0.463 

13 3 3 6 2.1 5.4 2.04 0.514 

14 30 3 9.4 3.4 9.46 2.04 -0.0514 

15 

16 

17 

18 Model Descriptions for likelihoods calculated 

19 

20 

21 Model A1: Yij = Mu(i) + e(ij) 

22 Var{e(ij)} = Sigma^2 

23 

24 Model A2: Yij = Mu(i) + e(ij) 

25 Var{e(ij)} = Sigma(i)^2 

26 

27 Model R: Yi = Mu + e(i) 

28 Var{e(i)} = Sigma^2 

29 

30 

31 Likelihoods of Interest 

32 

33 Model Log(likelihood) DF AIC 

34 A1 -10.652512 4 29.305023 

35 A2 -9.359638 6 30.719276 

36 fitted -10.899709 2 25.799418 

37 R -14.037484 2 32.074967 

38 

39 Test 1: Does response and/or variances differ among dose 

40 levels 

41 (A2 vs. R) 

42 Test 2: Are Variances Homogeneous (A1 vs A2) 

43 Test 3: Does the Model for the Mean Fit (A1 vs. fitted) 

44 

45 Tests of Interest 

46 

47 Test -2*log(Likelihood Ratio) Test df p-value 

48 

49 Test 1 9.35569 4 0.009299 

50 Test 2 2.58575 2 0.2745 

51 Test 3 0.494395 1 0.482 

52 

53 The p-value for Test 1 is less than .05. There appears 

54 to be a 

55 difference between response and/or variances among the 

56 dose levels. 

57 It seems appropriate to model the data 

58 

59 The p-value for Test 2 is greater than .05. A 

60 homogeneous variance 
D-158 DRAFT – DO NOT CITE OR QUOTE 

1 model appears to be appropriate here 

2 

3 

4 The p-value for Test 3 is greater than .05. The model 

5 chosen appears 

6 to adequately describe the data 

7 

8 

9 

10 Benchmark Dose Computation 

11 Specified effect = 7.6 

12 

13 Risk Type = Point risk 

14 

15 Confidence level = 0.95 

16 

17 BMD = 17.6423 

18 

19 

20 BMDL = 9.58925 

21 
E-1 DRAFT – DO NOT CITE OR QUOTE 

1 APPENDIX E. CALCULATION OF RPFs 

2 

3 
E-2 DRAFT – DO NOT CITE OR QUO

Table E-1. Dermal bioassays: RPF calculations for incidence data 

Relative potency calculation 

Record 

no. Reference

Tumor 

type(s) Sex PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose Dose units

Converted 

dose 

Converted 

dose 

units RPF Comment

Complete carcinogenicity studies 

600 Habs et al., 

1980 

Sum of Papilloma, carcinoma, sarcoma 

F BaP 0.1 0.91 pg/animal 1 High dose dropped 

 F BbF 0.1 3.8 pg/animal 0.24 13640 Cavalieri et 

al., 1983 

Papilloma, adenoma, carcinoma 

F BaP 0.1 5.3 nmol 0.001 mg 1 

 F CPcdP 0.1 47 nmol 0.011 mg 0.13 620 Hoffmann 

and Wynder, 1966 

Papillomas F BaP 0.1 0.0031 % 1 

 F DBaeP 0.1 0.0094 % 0.33 Toxicity resulted i

significant mortaliunrelated to tumor induction. 

 F DBaiP 0.1 0.0042 % 0.74  F DBaeF 0.1 0.0028 % 1.1 17660 Cavalieri et 

al., 1977 

Papilloma, keratoacanthoma, carcinoma 

F BaP 0.79 0.396 1 

 F AA 0.47 0.396 μmol/ 

application

0.109 mg/ 

application

0.55 

Initiation studies 

630 LaVoie et 

al., 1982 

Primarily squamous cell 

papilloma 

F BaP 0.85 30 μg/animal 1 

 F BbF 0.8 100 μg/animal 0.28 No model fit. Poin

estimate using incidose point closest incidence. 

 F BjF 0.85 209 μg/animal 0.14 High dose dropped  F BkF 0.85 1,163 μg/animal 0.03 18570 Hecht et 

al., 1974 

Unspecified F BaP 0.3 0.05 mg/animal 1 
E-3 DRAFT – DO NOT CITE OR QUO

Table E-1. Dermal bioassays: RPF calculations for incidence data 

Relative potency calculation 

Record 

no. Reference

Tumor 

type(s) Sex PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose Dose units

Converted 

dose 

Converted 

dose 

units RPF Comment

 F CH 0.58 1 mg/animal 0.10 21420 Slaga et 

al., 1980 

Papilloma F BaP 0.64 200 nmol 0.050 mg 1 

 F CH 0.71 2,000 nmol 0.457 mg 0.12 Not clear if BaP 

administered simultaneously. Cgroups pooled for 

 F DBahA 0.45 100 nmol 0.028 mg 1.27 15640 Raveh et 

al., 1982 

Papilloma F BaP 0.1 2.2 μg 1 

 F CPcdP 0.1 30 μg 0.07 620 Hoffmann 

and Wynder, 1966 

Papillomas F BaP 0.79 0.25 mg/animal 1 

 F DBaeF 0.57 0.25 mg/animal 0.73  F DBaeP 0.33 0.25 mg/animal 0.41  F DBahP 0.7 0.25 mg/animal 0.90  F DBaiP 0.36 0.25 mg/animal 0.45  F N23eP 0.25 0.25 mg/animal 0.32 13650 Cavalieri et 

al., 1981b 

Papillomas F BaP 0.33 0.2 μmol 0.050 mg 1 

 F CPcdP 0.23 0.6 μmol 0.136 mg 0.26 Mid dose borderlin

significant, high dtrend not; no modeRPF uses mid dose 

point estimate. 

15700 Rice et al., 

1988 

Unspecified F BaP 0.89 0.1 μmol 0.025 mg 1 

 F CH 0.89 0.5 μmol 0.114 mg 0.22 No model fit. Poin

estimate using poiclosest to BaP inci

 F CPdefC 0.88 0.22 μmol 0.053 mg 0.48  F BbcAC 0.89 2 μmol 0.481 mg 0.05 No model fit. Poin

estimate using poiclosest to BaP inci

24800 Nesnow et 

al., 1984 

Papilloma M BaP 0.67 200 nmol 0.050 mg 1 
E-4 DRAFT – DO NOT CITE OR QUO

Table E-1. Dermal bioassays: RPF calculations for incidence data 

Relative potency calculation 

Record 

no. Reference

Tumor 

type(s) Sex PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose Dose units

Converted 

dose 

Converted 

dose 

units RPF Comment

 M BeAC 0.67 393 nmol 0.099 mg 0.51 Three high doses d

due to plateau 

 M BlAC 0.67 50 nmol 0.013 mg 4.00 Three high doses d

due to plateau 

 F BaP 0.51 200 nmol 0.050 mg 1  F BeAC 0.51 195 nmol 0.049 mg 1.03 Two high doses dr

achieve model fit 

 F BlAC 0.51 30 nmol 0.008 mg 6.67 

1 
E-5 DRAFT – DO NOT CITE OR QUO

1 

Table E-2. Dermal bioassays: RPF calculations for multiplicity data 

Relative potency calculation 

Record no. Reference Tumor type(s) Sex PAH 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose 

Dose 

units 

Converted 

dose 

Converted 

dose units RPF Comments 

Complete carcinogenicity studies 

13640 Cavalieri et al., 

1983 

Papilloma, adenoma, carcinoma 

F BaP 1.5 20 nmol 0.0050 mg 1 Variance not reported

 F CPcdP 2.5 200 nmol 0.045 mg 0.18 Variance not reported 13650 Cavalieri et al., 

1981b 

Primarily squamous cell carcinoma 

US BaP 1.5 0.2 μmol 0.050 mg 1 

 US CPcdP 0.80 0.2 μmol 0.045 mg 0.59 Variance not reported 

Initiation studies

630 LaVoie et al., 

1982 

Primarily squamous cell 

papilloma 

F BaP 4.9 30 μg 1 

 F BbF 7.1 100 μg 0.43 Variance not reported  F BjF 7.2 1,000 μg 0.044 Variance not reported F BkF 2.8 1,000 μg 0.017 Variance not reported18570 Hecht et al., 

1974 

Unspecified F BaP 0.5 0.05 mg 1 

 F CH 1.0 1 mg 0.10 21420 Slaga et al., 

1980 

Papilloma F BaP 2.1 200 nmol 0.050 mg 1 

 F CH 1.5 2,000 nmol 0.46 mg 0.078 

 F DBahA 1.3 100 nmol 0.028 mg 1.1 15640 Raveh et al., 

1982 

Papilloma F BaP 1.1 10 μg 1 Variance not reported

 F CPcdP 0.7 200 μg 0.032 Variance not reported13650 Cavalieri et al., 

1981 

Papillomas F BaP 1.1 0.2 μmol 0.050 mg 1 

 F CPcdP 0.17 0.6 μmol 0.14 mg 0.060 Variance not reported 21410 Slaga et al., 

1978 

Papillomas F BaP 5.2 0.2 μmol 0.050 mg 1 

 F BaA 1.1 2 μmol 0.46 mg 0.023 16310 Weyand et al., 

1992 

Unspecified US BaP 4.0 0.01 μmol 0.0025 mg 1 

 US BjF 4.0 1 μmol 0.252 mg 0.010 Variance not reported 10200 El-Bayoumy et 

al., 1982 

Primarily squamous cell 

papilloma 

F BaP 7.0 0.05 mg 1 
E-6 DRAFT – DO NOT CITE OR QUO

Table E-2. Dermal bioassays: RPF calculations for multiplicity data 

Relative potency calculation 

Record no. Reference Tumor type(s) Sex PAH 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose 

Dose 

units 

Converted 

dose 

Converted 

dose units RPF Comments 

 F CH 7.6 1 mg 0.054 24300 Rice et al., 

1985 

Unspecified F BaP 7.9 0.3 mg 1 

 F CH 4.9 1 mg 0.18  F CPdefC 5.5 1 mg 0.21 13660 Cavalieri et al., 

1991 

Primarily 

papilloma 

F BaP Expt I 5.2 300 nmol 0.0757 mg 1 16 Wk experiment; variance not r

 F DBalP Expt I 6.8 33.3 nmol 0.010 mg 9.7 13660 Cavalieri et al., 

1991 

Primarily 

papilloma 

F BaP Expt II 3.4 100 nmol 0.0252 mg 1 27 Wk experiment; variance not r

 F DBalP Expt II 7.0 4 nmol 0.0012 mg 42 24800 Nesnow et al., 

1984 

Papillomas M BaP 1.4 200 nmol 0.050 mg 1 Variance not reported 

 M BeAC 1.3 250 nmol 0.063 mg 0.74 Variance not reported  M BlAC 1.4 50 nmol 0.013 mg 4.0 Variance not reported  F BaP 1.5 200 nmol 0.050 mg 1 Variance not reported  F BeAC 1.1 250 nmol 0.063 mg 0.58 Variance not reported  F BlAC 1.1 50 nmol 0.013 mg 2.9 Variance not reported 
E-7 DRAFT – DO NOT CITE OR QUO

1 

Table E-3. Intraperitoneal bioassays: RPF calculations for incidence data 

Relative potency calculation 

Record 

no. Reference

Target 

organ 

Tumor 

type(s) Sex PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units Converted dose 

Converted 

dose 

units RPF Com

17560 Busby et 

al., 1989 

Lung Adenoma, 

adenocarcinoma 

F BaP 0.68 59.5 μg 1 

 FA 0.26 257.6 μg 0.09 640 LaVoie et 

al., 1987 

Lung Adenoma M BaP 0.82 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

1 

 BjF 0.52 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

0.64 Do not 

liver or RPF be

 F BaP 0.64 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

1 

 BjF 0.22 1.1 umol/ 

mouse 

0.28 mg/ 

mouse 

0.35 Do not 

liver or RPF be

 Liver Adenoma, 

hepatoma 

M BaP 0.75 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

1 

 BbF 0.5 0.5 μmol/ 

mouse 

0.13 mg/ 

mouse 

1.50 Do not 

liver or RPF be

 BjF 0.49 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

0.66 Do not 

liver or RPF be

 Liver or 

lung 

Adenoma, hepatoma 

M BaP 0.75 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

1 

 BbF 0.51 0.5 μmol/ 

mouse 

0.13 mg/ 

mouse 

1.50 

 BjF 0.8 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

1.10 

 F BaP 0.64 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

1 

 BjF 0.22 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

0.35 

7510 LaVoie et 

al., 1994 

Lung Total M BaP 0.7 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

1 

 FA 0.7 22 μmol/ 

mouse 

4.45 mg/ 

mouse 

0.06 Do not 

male liis high
E-8 DRAFT – DO NOT CITE OR QUO

Table E-3. Intraperitoneal bioassays: RPF calculations for incidence data 

Relative potency calculation 

Record 

no. Reference

Target 

organ 

Tumor 

type(s) Sex PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units Converted dose 

Converted 

dose 

units RPF Com

 F BaP 0.83 1.1 μmol/ 

mouse 

0.28 mg/ 

mouse 

1 

 FA 0.83 17 μmol/ 

mouse 

3.44 mg/ 

mouse 

0.08 

 Liver Foci, adenoma, 

carcinoma 

M BaP 0.81 1.1 μmol 0.28 mg/ 

mouse 

1 

 FA 0.81 6.4 μmol 1.29 mg/ 

mouse 

0.21 

22510 Wislocki et 

al., 1986 

Liver Adenoma, 

carcinoma 

M BaP 0.44 560 nmol 0.14 mg 1 

 CH 0.44 3,339 nmol 0.76 mg 0.19 Using p

control

 BaA 0.77 2,800 nmol 0.64 mg 0.39  Lung Unspecified M BaP 0.3 560 nmol 0.14 mg 1  CH 0.3 5,601 nmol 1.28 mg 0.11 Do not 

male liis highUsing pcontrol

 F BaP 0.46 560 nmol 0.14 mg 1  BaA 0.16 2,800 nmol 0.64 mg 0.08 

1 2 
E-9 DRAFT – DO NOT CITE OR QUO

1 

Table E-4. Intraperitoneal bioassays: RPF calculations for multiplicity data 

RPF Calculation 

Record 

no. Reference 

Target 

organ(s) Tumor type(s) Sex PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose 

Dose 

units 

Converted 

dose 

Converted 

dose units RPF Com

17560 Busby et al., 

1989 

Lung Adenoma, 

adenocarcinoma 

F BaP 1.11 59.5 μg 1 

 FA 0.33 257.6 μg 0.069 7510 LaVoie et 

al., 1994 

Lung Total M BaP 4.13 1.1 μmol 0.28 mg 1 

 FA 0.95 17.30 μmol 3.50 mg 0.018 Do n

male RPF 

 F BaP 3.40 1.1 μmol 0.28 mg 1  FA 2.30 17.30 μmol 3.50 mg 0.054  Liver Foci, adenoma, 

carcinoma 

M BaP 4.12 1.1 μmol 0.28 mg 1 

 FA 1.45 3.46 μmol 0.700 mg 0.14 22510 Wislocki et 

al., 1986 

Liver Adenoma, 

carcinoma 

M BaP 1.36 560 nmol 0.141 mg 1 

 CH 0.93 2,800 nmol 0.639 mg 0.15 Using 

contr

 BaA 2.28 2,800 nmol 0.639 mg 0.37 13610 Busby et al., 

1984 

Lung Adenoma, 

carcinoma 

M BaP 4.28 0.28 mg 1 No m

 FA 4.28 9.99 mg 0.028  F BaP 3.56 0.28 mg 1 No m FA 3.56 32.28 mg 0.0086 24590 Nesnow et 

al., 1998b 

Lung NS M BaP 3.85 50 mg/kg 1 No m

 BbF 3.85 123 mg/kg 0.41 BMR 

respo

 CPcdP 4.15 50 mg/kg 1.1 No m DBahA 3.85 3.57 mg/kg 14 BMR 

respo

 DBalP 3.66 1.5 mg/kg 32 No m11190 Mass et al., 

1993 

Lung NS US BaP 5.05 100 mg/kg 1 No m

 BjAC 59.45 20 mg/kg 59 No m

2 
E-10 DRAFT – DO NOT CITE OR QUO

Table E-5. Lung implantation bioassays: RPF calculations (incidence data) 

Relative potency calculation

Record no. Reference Target organ(s) Tumor type(s) PAH BMR BMD 

Point 

estimate 

response 

Point 

estimate 

dose Dose units RPF Com

17940 Deutsch-Wenzel et al., 1983 Lung Sum carcinoma + sarcoma BaP 0.1 0.032 mg 1  AA 0.1 0.14 mg 0.24  BbF 0.1 0.33 mg 0.10  BghiP 0.1 3.5 mg 0.0092  BjF 0.1 1.0 mg 0.032  BkF 0.1 1.1 mg 0.031  IP 0.1 0.44 mg 0.074 22000 Wenzel-Hartung et al., 1990 Lung Carcinoma BaP 0.1 0.033 mg/ 

animal 

1 

 CH 0.1 0.85 mg/ 

animal 

0.038 

 BaP 0.57 0.20 mg/ 

animal 

1 

 DBahA 0.57 0.1 mg/ 

animal 

2.0 Sing

1 2 3 4 
E-11 DRAFT – DO NOT CITE OR QUO

1 

Table E-6. In vivo DNA adducts: RPF calculations 

Relative potency calculation 

Record 

no. Reference

Target 

organ(s)/route PAH AUC 

AUC vs. 

dose 

Point 

estimate 

extra risk 

response

Point 

estimate 

dose Dose units Converted dose 

Converted dose

units RPF Comm

6210 Arif et al., 

1997 

Sum of adducts in mammary gland, lung, heart, 

pancreas, bladder, 

liver 

BaP 325 0.25 μmol/ 

mammary gland 

0.063 mg/ 

mammary gland

1 

 DBalP 2,245 0.25 μmol/ 

mammary gland 

0.076 mg/mammary 

gland 

5.8 

17630 Cavalieri et 

al., 1981a 

Skin 4-hr BaP 16 0.2 μmol/animal 0.050 mg/animal 1 Higher of 

measured 

 ACEP 2.2 0.2 μmol/animal 0.046 mg/animal 0.15 Higher of 

measured 

 CPcdP 8.8 0.2 μmol/animal 0.045 mg/animal 0.61 Higher of 

measured 

18810 Hughes and 

Phillips, 1990 

Sum of skin and lung 

BaP 9 1 μmol 0.25 mg 1 RPFs base

peaks; dig

not possibPeaks reacdifferent t

postdosing

 DBaeP cannot 

determine

1 μmol NA 

 DBahP 3.2 1 μmol 0.30 mg 0.30  DBaiP 0.85 1 μmol 0.30 mg 0.079  DBalP 65 1 μmol 0.30 mg 6.0 11190 Mass et al., 

1993 

Lung BaP 470 mg/kg 1 

 BjAC 464 mg/kg 0.99 Ratio of sl

AUC vs. dBjAC plot curvature 

8010 Nesnow et 

al., 1993b 

Total of lung, liver, and peripheral 

blood lymphocytes 

BaP 52,084 100 mg/kg 1 

 BbF 11,314 100 mg/kg 0.22 Ratio of (s

AUCs)/do
E-12 DRAFT – DO NOT CITE OR QUO

Table E-6. In vivo DNA adducts: RPF calculations 

Relative potency calculation 

Record 

no. Reference

Target 

organ(s)/route PAH AUC 

AUC vs. 

dose 

Point 

estimate 

extra risk 

response

Point 

estimate 

dose Dose units Converted dose 

Converted dose

units RPF Comm

24590 Nesnow et 

al., 1998b 

Lung BaP 113 mg/kg 1 Ratio of sl

AUC vs. dreported bauthors 

 BbF 38 mg/kg 0.33  CPcdP 148 mg/kg 1.3  DBahA 219 mg/kg 1.9  DBalP 1,390 mg/kg 12 22810 Phillips et 

al., 1979 

Skin BaP 27 1 0.25 mg/animal 1 Ratio of p

levels. Pereached at different t

 DBacA 10 1 μmol/animal 0.28 mg/animal 0.34  DBahA 15 1 μmol/animal 0.28 mg/animal 0.50 24790 Kligerman 

et al., 2002

Mouse peripheral 

blood lymphocytes/

intraperitoneal 

BaP 4,186 100 mg/kg 1 Ratio of si

measure o

postdosing 

 BaA 93 100 mg/kg 0.022  BbF 516 100 mg/kg 0.12  CH 81 100 mg/kg 0.019  Mouse peripheral 

blood lymphocytes/

gavage 

BaP 143 100 mg/kg 1 

 BaA 32 100 mg/kg 0.22  BbF 39 100 mg/kg 0.27  CH 37 100 mg/kg 0.26  Rat peripheral 

blood lymphocytes/ 

intraperitoneal 

BaP 755 100 mg/kg 1 

 BaA 38 100 mg/kg 0.05  BbF 63 100 mg/kg 0.083  CH 24 100 mg/kg 0.032  Rat peripheral 

blood lymphocytes/

gavage 

BaP 177 100 mg/kg 1 

 BaA 20 100 mg/kg 0.11  BbF 17 100 mg/kg 0.1  CH 10 100 mg/kg 0.056 

1 
E-13 DRAFT – DO NOT CITE OR QUO

1 

Table E-7. In vivo clastogenicity or sister chromatid exchange: RPF calculation 

Relative potency calculation 

Record 

no. Reference Route Endpoint 

Data type: quantal 

or continuous PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units RPF Commen

24740 Allen et al., 

1999 

Intraperitoneal Micronucleated 

polychromatic 

erythrocytes in bone marrow (A/J mouse)

Q BaP 0.0086 200 mg/kg 1 

 DBalP 0.0013 1.5 mg/kg 20 Model won't pr

BaP BMR. RPon peak 

 Intraperitoneal Micronucleated 

polychromatic 

erythrocytes in 

peripheral blood 

(A/J mouse) 

Q BaP 0.0067 200 mg/kg 1 

 DBalP 0.0015 6 mg/kg 7.5 Model won't pr

BaP BMR. RPon peak 

 Intraperitoneal Micronucleated 

polychromatic 

erythrocytes in bone marrow (p53 wt mouse) 

Q BaP 0.0019 200 mg/kg 1 

 DBalP 0.0042 12 mg/kg 37 Model won't pr

BaP BMR. RPon peak 

 Intraperitoneal Micronucleated 

polychromatic 

erythrocytes in peripheral blood (p53 wt mouse) 

Q BaP 0.0022 200 mg/kg 1 

 DBalP 0.0011 18 mg/kg 5.6 BMD doesn't re

selected BMR. 

based on peak. 

14270 He and Baker, 

1991 

Dermal micronuclei Q BaP 0.064 50 μg/anim

al 

1 No model fit. R

based on peak. 

 CH 0.05 500 μg/anim

al 

0.078 No model fit. R

based on peak. 
E-14 DRAFT – DO NOT CITE OR QUO

Table E-7. In vivo clastogenicity or sister chromatid exchange: RPF calculation 

Relative potency calculation 

Record 

no. Reference Route Endpoint 

Data type: quantal 

or continuous PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units RPF Commen

17190 Bayer, 1978 Intraperitoneal Sister chromatid 

exchanges 

C BaP 4.2 100 mg/kg 1 No model fit. R

based on peak. 

 PH 0.9 100 mg/kg 0.21 No model fit. R

based on peak. 

20950 RoszinskyKocher et al., 

1979 

Intraperitoneal Sister chromatid 

exchanges 

C BaP 6.7 900 mg/kg 1 

 DBahA 1 900 mg/kg 0.15  CH 1.2 900 mg/kg 0.18  PH 1.6 900 mg/kg 0.24  BeP 1.6 900 mg/kg 0.24  BbF 1.7 900 mg/kg 0.25  BaA 2.2 900 mg/kg 0.33 24720 Kligerman et 

al., 1986 

Gavage Sister chromatid 

exchanges 

C BaP 8 63 mg/kg 1 No SD for cont

 BlAC 16 126 mg/kg 1.1 No SD for cont

based on lowes

approaching pe

24790 Kligerman et 

al., 2002 

Intraperitoneal Sister chromatid 

exchanges 

C BaP 12.42 100 mg/kg 1 

 BaA 6.01 100 mg/kg 0.48  BbF 13.46 100 mg/kg 1.1  CH 3.17 100 mg/kg 0.26  Gavage Sister chromatid 

exchanges 

C BaP 6.79 100 mg/kg 1 

 BaA 2.26 100 mg/kg 0.33  Gavage Micronuclei Q BaP 0.0025 100 mg/kg 1  BbF 0.0017 100 mg/kg 0.68 

1 2 
E-15 DRAFT – DO NOT CITE OR QUO

1 

Table E-8. In vitro bacterial mutagenicity: RPF calculations 

Relative potency calculation

Record 

no. Reference PAH 

Data type: quantal or 

continuous BMR BMD Slope

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units Converted dose Converted dose units RPF Commen

17030 Andrews et 

al., 1978 

BaP C 1,531 250 μg 1 

 DBacA C 2,807 10 μg 46  DBajA C 693 10 μg 11  DBahA C 467 25 μg 3  AA C 1,645 250 μg 1.1  BghiP C 642 100 μg 1  BeP C 492 1,000 μg 0.08 23830 Baker et al., 

1980 

BaP C 1,144 2.5 μg/plate 1 

 DBaiP C 603 5 μg/plate 0.26  BaA C 813 10 μg/plate 0.18  DBacA C 1,604 2.5 μg/plate 1.4  DBahA C 1,197 5 μg/plate 0.52 23660 Bartsch et 

al., 1980 

BaP C 29,000 0.027 μmol/plate 0.007 mg/plate 1 

 BaA C 6,000 0.067 μmol/plate 0.015 mg/plate 0.092 17380 Bos et al., 

1988 

BaP C 739 7.5 μg/plate 1 RPF based on pe

response. BaP rewell above range other data sets; mrequired droppindoses but not appgiven BMR targe

 PH C 155 25 μg/plate 0.063  Pyr C 193 25 μg/plate 0.078 17590 Carver et al., 

1986 

BaP C 895 50 μg/plate 1 Continuous data

RPF based on pelowest dose appr

peak 

 BaA C 1,123 50 μg/plate 1.3  BghiF C 845 50 μg/plate 0.94  Pery C 853 10 μg/plate 4.8 17630 Cavalieri et 

al., 1981a 

BaP Q 0.00126 60 μM 15.1 mg/L 1 RPF based on pe

model fit 

 CPcdP Q 0.0013 40 μM 9.1 mg/L 1.7 RPF based on pe

model fit 
E-16 DRAFT – DO NOT CITE OR QUO

Table E-8. In vitro bacterial mutagenicity: RPF calculations 

Relative potency calculation

Record 

no. Reference PAH 

Data type: quantal or 

continuous BMR BMD Slope

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units Converted dose Converted dose units RPF Commen

 ACEP Q 0.0005 120 μM 27.4 mg/L 0.22 RPF based on pe

BMD doesn't coiwith selected BM

9620 Chang et al., 

2002 

BaP C 2,217 5 μg/plate 1 Continuous data

RPF based on pelowest dose apprpeak 

 BghiF C 1,304 5 μg/plate 0.59  BcPH C 717 10 μg/plate 0.16 24030 De Flora et 

al., 1984 

BaP NA 185 revertants/nmol 733,196 revertants/mg 1 RPFs based on p

estimates as repoauthors 

 BaA NA 12 revertants/nmol 52,565 revertants/mg 0.072  BeP 1.6 revertants/nmol 6,341 revertants/mg 0.009  Pery NA 21 revertants/nmol 83,229 revertants/mg 0.11 18050 Eisenstadt 

and Gold, 1978 

BaP C 1,705 2 μg 1 Uses S9 level wi

BaP response; Cmax at much low

 CPcdP C 134 1 μg 0.16 18180 Florin et al., 

1980 

BaP C 255 0.003 μmol/plate 0.001 mg/plate 1 TA100 

 BaA C 326 0.1 μmol/plate 0.023 mg/plate 0.042  CH C 196 0.005 μmol/plate 0.001 mg/plate 0.51  BaP C 235 0.003 μmol/plate 0.001 mg/plate 1 TA 98  CO C 82 0.07 μmol/plate 0.021 mg/plate 0.013 Pery C 91 0.025 μmol/plate 0.006 mg/plate 0.046 24080 Gibson et 

al., 1978 

BaP C 35 300 μg/plate 1 Continuous data

RPF based on pelowest dose appr

peak. Metabolic 

activation by gamradiation. 

 BaA C 6.4 250 μg/plate 0.22  BghiP C 4.2 400 μg/plate 0.090  CH C 6.1 500 μg/plate 0.1 Lowest dose app

peak 

 FE C 2.2 360 μg/plate 0.052 
E-17 DRAFT – DO NOT CITE OR QUO

Table E-8. In vitro bacterial mutagenicity: RPF calculations 

Relative potency calculation

Record 

no. Reference PAH 

Data type: quantal or 

continuous BMR BMD Slope

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units Converted dose Converted dose units RPF Commen

 Pyr C 28 160 μg/plate 1.5 14080 Gold and 

Eisenstadt, 1980 

BaP C 1,103 4 nmol 0.001 mg 1 

 CPcdP C 281 4 nmol 0.001 mg 0.28 18650 Hermann, 

1981 

BaP NA 100 revertants/nmol 396,322 revertants/mg 1 RPFs based on p

estimates as repoauthors 

 AA NA 62 revertants/nmol 224,394 revertants/mg 0.57  BaA NA 4 revertants/nmol 17,522 revertants/mg 0.044  BbA NA 8 revertants/nmol 35,043 revertants/mg 0.088  BbF NA 15 revertants/nmol 59,448 revertants/mg 0.15  BeP NA 15 revertants/nmol 59,449 revertants/mg 0.15  CH NA 2 revertants/nmol 8,761 revertants/mg 0.022  CO NA 60 revertants/nmol 199,761 revertants/mg 0.50  DBacA NA 42 revertants/nmol 150,888 revertants/mg 0.38  DBahA NA 8 revertants/nmol 28,743 revertants/mg 0.073  DBaiP NA 38 revertants/nmol 125,661 revertants/mg 0.32  DBalP NA 21 revertants/nmol 69,451 revertants/mg 0.18  FA NA 3 revertants/nmol 14,832 revertants/mg 0.037  Pery NA 31 revertants/nmol 122,862 revertants/mg 0.31  Tphen NA 13 revertants/nmol 56,944 revertants/mg 0.14 10670 Johnsen et 

al., 1997 

BaP C 128 10 μg/plate 1 

 BjAC C 192 10 μg/plate 1.5 RPF based on pe

model fit 

 BlAC C 204 10 μg/plate 1.6 RPF based on pe

model fit 

19000 Kaden et al., 

1979 

BaP NA 1 RPFs as reported 

authors 

 AA NA 0.08  AN NA 0.01  ANL NA 0.07  BaA NA 0.14  BbFE NA 0.08  BeP NA 0.11  BghiP NA 0.08  CH NA 0.2 
E-18 DRAFT – DO NOT CITE OR QUO

Table E-8. In vitro bacterial mutagenicity: RPF calculations 

Relative potency calculation

Record 

no. Reference PAH 

Data type: quantal or 

continuous BMR BMD Slope

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units Converted dose Converted dose units RPF Commen

 CPcdP NA 1.5  DBacA NA 0.77  DBahA NA 0.08  DBbeF NA 0.88  FA NA 1  Pery NA 6  Pyr NA 0.07  Tphen NA 0.07 24680 Lafleur et 

al., 1993 

BaP Q 0.00026 8 μg/mL 1 RPF based on pe

BMD doesn't coiwith selected BM

 BghiF Q 0.00044 10 μg/mL 1.4  CPcdP Q 0.00048 8 μg/mL 1.9  CPhiACEA Q 0.00059 4 μg/mL 4.6  CPhiAPA Q 0.00017 100 μg/mL 0.05  ACEA Q 0.00059 35 μg/mL 0.53  APA Q 0.00026 30 μg/mL 0.27 19320 LaVoie et 

al., 1979 

BaP C 480 20 μg 1 Continuous data

RPF based on pelowest dose appr

peak 

 BeP C 20 10 μg 0.08  Pery C 70 20 μg 0.15 23650 McCann et 

al., 1975 

BaP NA 121 revertants/nmol 479,550 revertants/mg 1 RPFs based on p

estimates as repoauthors; authors that dose-responnonlinear 

 BaA NA 11 revertants/nmol 48,184 revertants/mg 0.10  BeP NA 0.6 revertants/nmol 2,378 revertants/mg 0.005  CH NA 38 revertants/nmol 166,455 revertants/mg 0.35  DBacA NA 175 revertants/nmol 628,698 revertants/mg 1.3  DBahA NA 11 revertants/nmol 39,521 revertants/mg 0.082  DBaiP NA 20 revertants/nmol 66,138 revertants/mg 0.14 20220 Pahlman 

and Pelkonen, 1987 

BaP NA 272 revertants/mg 1,077,996 revertants/mg 1 RPFs based on p

estimates as repoauthors 

 BaA NA 10 revertants/mg 43,804 revertants/mg 0.041 
E-19 DRAFT – DO NOT CITE OR QUO

Table E-8. In vitro bacterial mutagenicity: RPF calculations 

Relative potency calculation

Record 

no. Reference PAH 

Data type: quantal or 

continuous BMR BMD Slope

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units Converted dose Converted dose units RPF Commen

 CH NA 9.7 revertants/mg 42,490 revertants/mg 0.039  DBacA NA 35 revertants/mg 125,740 revertants/mg 0.12  DBahA NA 4 revertants/mg 14,371 revertants/mg 0.013  Tphen NA 4 revertants/mg 17,521 revertants/mg 0.016 20450 Phillipson 

and Ioannides, 1989 

BaP C 119 10 μg/plate 1 No SD; RPFs ba

peak or lowest d

approaching pea

 BaA C 110 20 μg/plate 0.46  DBaiP C 65 20 μg/plate 0.27  DBahA C 51 10 μg/plate 0.43 21000 Sakai et al., 

1985 

BaP C 1,565 10 μg 1 No SD; RPFs ba

peak or lowest d

approaching pea

 FE C 65 5 μg 0.083  AC C 320 10 μg 0.2  PH C 345 10 μg 0.22  FA C 835 10 μg 0.53  CH C 638 10 μg 0.41  Pyr C 2,400 10 μg 1.5  BeP C 923 10 μg 0.59  Pery C 2,607 4 μg 4.2  BghiP C 814 20 μg 0.26  CO C 223 10 μg 0.14 11860 Sangaiah et 

al., 1983 

BaP C 384 6 μg/plate 1 No SD; RPFs ba

peak or lowest d

approaching pea

 BjAC C 940 10 μg/plate 1.4 21360 Simmon, 

1979a 

BaP C 1,141 5 μg 1 

 BaA C 280 50 μg 0.025  BeP C 57 50 μg 0.005 21640 Teranishi et 

al., 1975 

BaP C 39 50 μg/plate 1 

 DBaiP C 64 50 μg/plate 1.6  BaP 254 50 μg/plate 1  DBaeP 63 50 μg/plate 0.25 
E-20 DRAFT – DO NOT CITE OR QUO

Table E-8. In vitro bacterial mutagenicity: RPF calculations 

Relative potency calculation

Record 

no. Reference PAH 

Data type: quantal or 

continuous BMR BMD Slope

Point 

estimate 

extra risk 

response 

Point estimate dose Dose units Converted dose Converted dose units RPF Commen

16180 Utesch et 

al., 1987 

BaP C 839 6 μg/plate 1 No SD; RPF bas

peak or lowest d

approaching pea

 BaA C 3,347 25 μg/plate 1 16440 Wood et al., 

1980 

BaP C 99 15 μg/plate 1 No SD; RPF bas

peak or lowest d

approaching pea

 CPcdP C 685 15 μg/plate 6.9 

1 
E-21 DRAFT – DO NOT CITE OR QUO

1 

Table E-9. In vitro mammalian mutagenicity: RPF calculations 

Relative potency calculation

Record 

no. Reference PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate dose Dose units 

Converted 

dose 

Converted 

dose 

units RPF Comment

16920 Amacher and 

Paillet, 1982 

BaP 0.00023 10 μg/mL 1 No model fit. RPF 

on peak 

 BaA 0.000068 10 μg/mL 0.3 No model fit. RPF 

on peak 

16940 Amacher and 

Turner, 1980 

BaP 0.00025 1.25 × 10-5 M 3.15 mg/L 1 Control w/o S9 tre

 BaA 0.00027 3.22 × 10-5 M 7.35 mg/L 0.46 16910 Amacher et al., 

1980 

BaP 0.00033 3.96 × 10-5 M 9.99 mg/L 1 No model fit. RPF 

on peak 

 BaA 0.00007 4.3 × 10-5 M 9.82 mg/L 0.22 BMD doesn't coinc

selected BMR; RPon peak 

17140 Barfknecht et al., 

1982 

BaP 0.00001 1.8 μM 0.45 mg/L 1 

 BaA 0.00001 23 μM 5.25 mg/L 0.09  CH 0.00001 16 μM 3.65 mg/L 0.12  CPcdP 0.0000083 23 μM 5.20 mg/L 0.07 BMD doesn't coinc

selected BMR; RPon response closesBMR of 0.00001 

 FA 0.00001 3.9 μM 0.79 mg/L 0.58  Tphen 0.00001 54 μM 12.33 mg/L 0.04 24670 Durant et al., 1999 BaP 0.00017 1,000 ng/mL 1 RPF based on peak 

response. Single dresponse at upper eabove data range fother data sets; morequired dropping doses but not approgiven BMR target response level 

 BaPery 0.00018 100 ng/mL 11  BbPery 0.000036 100 ng/mL 2.2  DBaeF 0.00017 100 ng/mL 10  DBafF 0.00017 1,000 ng/mL 1  DBahP 0.000061 100 ng/mL 3.7 
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Table E-9. In vitro mammalian mutagenicity: RPF calculations 

Relative potency calculation

Record 

no. Reference PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate dose Dose units 

Converted 

dose 

Converted 

dose 

units RPF Comment

 DBaiP 0.00013 100 ng/mL 7.8  DBelP 0.000034 1,000 ng/mL 0.21  N23aP 0.000073 100 ng/mL 4.4  N23eP 0.000079 1,000 ng/mL 0.48 14250 Hass et al., 1982 BaP 0.00026 0.3 μg/mL 1 No model fit. Resp

low dose (approach

peak) 

 DBaiP 0.0012 0.3 μg/mL 4.6 No model fit. RPF 

on peak 

 DBahP 0.00066 0.3 μg/mL 2.5 No model fit. RPF 

on peak 

18740 Huberman and 

Sachs, 1976 

BaP 0.0042 1 μg/mL 1 

 DBacA 0.00016 1 μg/mL 0.04  DBahA 0.00011 1 μg/mL 0.03 18990 Jotz and Mitchell, 

1981 

BaP 0.00014 4.5 μg/mL 1 With metabolic act

 Pyr 0.000034 11 μg/mL 0.1 With metabolic act24720 Kligerman et al., 

1986 

BaP 0.00047 4 nmol/mL 0.001 mg/mL 1 No model fit. RPF 

on peak 

 BlAC 0.00028 5 nmol/mL 0.0013 mg/mL 0.48 No model fit. RPF 

on peak 

19180 Krahn and 

Heidelberger, 1977

BaP 0.00012 15.9 nmol/mL 0.004 mg/mL 1 3-MC S9; 40% sur

 BaA 0.00005 46.5 nmol/mL 0.011 mg/mL 0.16 3-MC S9; 40% sur24680 Lafleur et al., 1993 BaP 0.000024 0.2 μg/mL 1 No model fit.  ACEA 0.000013 3 μg/mL 0.037 No model fit.  CPcdP 0.000015 2 μg/mL 0.061 No model fit.  CPhiACEA 0.000022 0.3 μg/mL 0.62 No model fit. 7550 Li and Lin, 1996 BaP 0.00003 10 ng/mL 1  BaA 0.000036 10 ng/mL 1.2 11450 Nesnow et al., 

1984 

BaP 0.00019 5 μg/mL 1 
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Table E-9. In vitro mammalian mutagenicity: RPF calculations 

Relative potency calculation

Record 

no. Reference PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate dose Dose units 

Converted 

dose 

Converted 

dose 

units RPF Comment

 BeAC 0.00042 5 μg/mL 2.2 No model fit; RPF 

on lowest dose approaching peak 

 BjAC 0.00025 5 μg/mL 1.3 No model fit; RPF 

on lowest dose approaching peak 

 BlAC 0.00044 2.5 μg/mL 4.6 No model fit; RPF 

on lowest dose approaching peak 

15630 Raveh and 

Huberman, 1983 

BaP 0.0001 0.11 μg/mL 1 

 CPcdP 0.0001 0.58 μg/mL 0.18 Uses QL; MS didn

converge 

15640 Raveh et al., 1982 BaP 0.00001 0.16 μg/mL 1 Uses QL, hi dose d

MS didn't fit 

 CPcdP 0.00001 1.1 μg/mL 0.14 Uses QL; MS didn

converge 

21410 Slaga et al., 1978 BaP 0.0001 0.048 μM 0.012 mg/L 1  BaA 0.0001 32 μM 7.3 mg/L 0.0016

58 

16190 Vaca et al., 1992 BaP 0.00027 10 μM 2.5 mg/L 1 BMD doesn't coinc

selected BMR; RPon peak 

 FA 0.00021 10 μM 2.02 mg/L 0.97 BMD doesn't coinc

selected BMR; RPon peak 

21900 Wangenheim and 

Bolcsfoldi, 1988 

BaP 0.0008 0.00001 mol/L 2.5 mg/L 1 BMD doesn't coinc

selected BMR; RPon peak 

 FE 0.000086 0.00012 mol/L 19.9 mg/L 0.014 BMD doesn't coinc

selected BMR; RPon peak 

 Pyr 0.00053 0.00003 mol/L 6.1 mg/L 0.28 BMD doesn't coinc

selected BMR; RPon peak 

1 
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1 

Table E-10. In vitro morphological/malignant transformation: RPF calculation 

Relative potency calculation 

Record 

no. Reference PAH 

Data type: quantal or 

continuous BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimat

e dose 

Slope of 

dose 

response 

curve 

Dose 

units Converted dose

Converted dose

units RPF Com

17610 Casto, 1979 BaP Q 0.00001 0.1 μg/mL 1  DBahA Q 0.00001 2.5 μg/mL 0.042 17970 DiPaolo et 

al., 1969 

BaP Q 0.058 10 μg/mL 1 

 DBahA Q 0.031 10 μg/mL 0.54  BaA Q 0.011 10 μg/mL 0.18  BeP Q 0.0058 10 μg/mL 0.1  DBacA Q 0.011 10 μg/mL 0.19 18080 Emura et al., 

1980 

BaP Expt I Q 0.001 0.044 μg/mL 1 

 BbF Q 0.001 0.75 μg/mL 0.059  BaA Q 0.001 0.85 μg/mL 0.052  BaP Expt II Q 0.001 0.046 μg/mL 1  IP Q 0.001 0.82 μg/mL 0.056 14130 Greb et al., 

1980 

BaP NA 277 %/mmol 1.10 %/mg 1 Relative 

transform

potencie

reportedare ratio 

potencie

 BaA NA 13.9 %/mmol 0.061 %/mg 0.055  BbF NA 11.5 %/mmol 0.046 %/mg 0.042  BeP NA 3.1 %/mmol 0.012 %/mg 0.011  CH NA 37 %/mmol 0.16 %/mg 0.15  DBahA NA 0.3 %/mmol 0.001 %/mg 0.00098214640 Krolewski 

et al., 1986 

BaP Q 0.0055 5 μM 1.3 mg/L 1 

 CPcdP Q 0.0017 5 μM 1.1 mg/L 0.34 14700 Laaksonen 

et al., 1983 

BaP Q 0.000009 10 μM 2.5 mg/L 1 RPF bas

peak. In

dose-resrelations

possibly 

cytotoxi
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Table E-10. In vitro morphological/malignant transformation: RPF calculation 

Relative potency calculation 

Record 

no. Reference PAH 

Data type: quantal or 

continuous BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimat

e dose 

Slope of 

dose 

response 

curve 

Dose 

units Converted dose

Converted dose

units RPF Com

 BaA Q 0.000018 11 μM 2.5 mg/L 2.0 RPF bas

peak. In

dose-resrelations

possibly 

cytotoxi

14850 Lubet et al., 

1983 

BaP Q 0.1 1.9 μg/mL 1 

 BeP Q 0.1 41 μg/mL 0.046 24710 Mohapatra 

et al., 1987 

BaP 0.92 1 μg/mL 1 

 BjAC Q 0.92 0.93 μg/mL 1.1  BaP 0.83 1 μg/mL 1  BlAC Q 0.83 7.5 μg/mL 0.13  BaP 0.86 1 μg/mL 1  BeAC Q 0.86 18 μg/mL 0.056 24700 Nesnow et 

al., 1990 

BaP C 47 10 μg/mL 1 

 BlAC C 120 10 μg/mL 2.5 Based o

responsefor cont

7980 Nesnow et 

al., 1997 

BaP C 2.5 4 μM 1.01 mg/L 1 

 DBalP C 1.7 0.33 μM 0.10 mg/L 6.9 Based o

responsefor cont

7990 Nesnow et 

al., 1994 

BaP C 0.94 1 μg/mL 1 

 DBahA C 0.37 1 μg/mL 0.39 Based o

responsecontinuolinear m

8000 Nesnow et 

al., 1993a 

BaP C 1.4 3 μg/mL 1 

 DBkmnoAPH C 1.1 5 μg/mL 0.47 Based o

responsefor cont

23720 Pienta et al., 

1977 

BaP Q 0.01 15 μg/mL 1 High do

dropped 
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Table E-10. In vitro morphological/malignant transformation: RPF calculation 

Relative potency calculation 

Record 

no. Reference PAH 

Data type: quantal or 

continuous BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimat

e dose 

Slope of 

dose 

response 

curve 

Dose 

units Converted dose

Converted dose

units RPF Com

 BaA Q 0.01 8.2 μg/mL 1.9 Caution

changinin region BMR 

 DBahA Q 0.01 0.4 μg/mL 34 Two hig

doses dr

1 
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1 

Table E-11. In vitro DNA adducts: RPF calculations

a

Relative potency calculation 

Record no. Reference PAH 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose Dose units 

Converted 

dose 

Converted 

dose 

units RPF Comments 

16890 Allen and Coombs, 1980 BaP 7.5 0.24 μg/mL 1 Nuclear DNA  BaA 0.44 0.64 μg/mL 0.021  BaP 413 0.24 μg/mL 1 Mitochondrial DNA  BaA 104 0.64 μg/mL 0.092 6300 Binkova et al., 2000 BaP 258 1 μM 0.25 mg/L 1  DBalP 2,317 0.1 μM 0.03 mg/L 75 9510 Bryla and Weyand, 1992 BaP 5.5 600 nmol 0.15 mg 1 Light conditions  BaA 1 600 nmol 0.14 mg 0.20  DBacA 1.8 600 nmol 0.17 mg 0.30 22800 Grover and Sims, 1968 BaP 1.4 5 μg 1  DBahA 0.44 5 μg 0.31  DBacA 0.56 5 μg 0.40  BaA 0.7 5 μg 0.50  Pyr 0.31 5 μg 0.22  PH 0.05 5 μg 0.040 10670 Johnsen et al., 1997 BaP 0.05 30 μg/mL 1 Clara cells  BjAC 0.15 30 μg/mL 3  BlAC 0.24 30 μg/mL 4.8  BaP 0.02 30 μg/mL 1 Type 2 cells  BjAC 0.06 30 μg/mL 3  BlAC 0.03 30 μg/mL 1.5 10660 Johnsen et al., 1998 BaP 0.33 30 μg/mL 1 Human lymphocytes  BjAC 0.11 30 μg/mL 0.33  BlAC 1.1 30 μg/mL 3.3  BaP 0.24 30 μg/mL 1 HL-60 cells  BjAC 0.15 30 μg/mL 0.62  BlAC 0.94 30 μg/mL 3.9 7870 Melendez-Colon et al., 2000 BaP 34 2 μM 0.50 mg/L 1  DBalP 348 2 μM 0.60 mg/L 8.5 21200 Segerback and Vodicka, 1993 BaP 15 100 mM 25,232 mg/L 1  BaA 30 100 mM 22,829 mg/L 2.2  BbF 3.7 100 mM 25,232 mg/L 0.25  BghiP 0.5 100 mM 27,634 mg/L 0.03  CH 50 100 mM 22,829 mg/L 3.7  DBahA 2.8 100 mM 27,833 mg/L 0.17 
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Table E-11. In vitro DNA adducts: RPF calculations

a

Relative potency calculation 

Record no. Reference PAH 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose Dose units 

Converted 

dose 

Converted 

dose 

units RPF Comments 

 FA 1.5 100 mM 20,226 mg/L 0.12  Pyr 0.14 100 mM 20,226 mg/L 0.012 aAll RPFs are point estimates based on peak response as adequate model fit was not achieved for any multi-dose dataset. 

No control data were available for any of these studies. 

1 2 3 4 5 
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1 

Table E-12. In vitro DNA damage: RPF calculations

Relative potency calculation 

Record 

no. Reference PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose 

Slope of 

dose 

response 

curve Dose units

Converted 

dose 

Converted 

dose 

units RPF Comments 

16840 Agrelo and 

Amos, 1981

BaP 2,093 10 μg/mL 1 Control responses for BaP and Pyr 

by 10x. 

 Pyr 548 100 μg/mL 0.026 RPF based on peak; continuous da

without SD 

23790 Ichinotsubo 

et al., 1977 

BaP 6 70 μg/well 1 

 DBaiP 10 600 μg/well 0.19  DBahA 10 25 μg/well 4.7 10660 Johnsen et 

al., 1998 

BaP 7.9 3 μg/mL 1 Human lymphocytes. No model f

Lowest response point estimate 

 BjAC 7.6 18 μg/mL 0.16 Human lymphocytes. BMR is BaP 

estimate response 

 BlAC 4.9 30 μg/mL 0.062 Human lymphocytes. No model f

Response point estimate closest to response. 

 BaP 5.4 30 μg/mL 1 HL-60 cells  BjAC 1.8 30 μg/mL 0.33 HL-60 cells  BlAC 3.8 30 μg/mL 0.7 HL-60 cells 19740 Martin et 

al., 1978 

BaP 210 1 × 10-5 M 2.5 mg/L 1 Increase over background 

 BaA 59 1 × 10-7 M 0.023 mg/L 31  BeP 256 1 × 10-6 M 0.25 mg/L 12  DBacA 97 1 × 10-5 M 2.8 mg/L 0.42  DBahA 96 1 × 10-5 M 2.8 mg/L 0.41 19830 MerschSunderman

n et al., 1992 

BaP 0.61 μg/assay 1 SOSIP - slope of SOS induction d

response curve as reported 

 AA 0.14 μg/assay 0.23  BaA 0.1 μg/assay 0.17  BbF 0.045 μg/assay 0.074  BghiF 0.34 μg/assay 0.56  BjF 0.25 μg/assay 0.42  BbFE 0.024 μg/assay 0.04  BghiP 0.033 μg/assay 0.055  BeP 0.032 μg/assay 0.053  CH 0.22 μg/assay 0.37  DBacA 0.10 μg/assay 0.17 
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Table E-12. In vitro DNA damage: RPF calculations

Relative potency calculation 

Record 

no. Reference PAH BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose 

Slope of 

dose 

response 

curve Dose units

Converted 

dose 

Converted 

dose 

units RPF Comments 

 DBahA 0.039 μg/assay 0.064  DBalP 2.1 μg/assay 3.5  DBahP 0.12 μg/assay 0.19  DBaiP 0.17 μg/assay 0.29  FA 0.41 μg/assay 0.68  IP 0.036 μg/assay 0.06  PH 0.053 μg/assay 0.088  Tphen 0.26 μg/assay 0.43 20810 Robinson 

and Mitchell, 1981 

BaP 89 10 μg/mL 1 

 Pyr 63 7.2 μg/mL 0.98 20940 Rossman et 

al., 1991 

BaP 10.4 12.5 μg/mL 1 Enhancement over background 

 AC 4.8 12.5 μg/mL 0.46  DBacA 8 1.44 μg/mL 6.7  DBahA 4 2 μg/mL 2.4  PH 4.5 25 μg/mL 0.22 21730 Tong et al., 

1981b 

BaP 65.5 0.001 M 252 mg/L 1 

 BaA 17.1 0.0005 M 114 mg/L 0.58 Based on peak response; no mode

1 2 
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1 

Table E-13. In vitro clastogenicity or sister chromatid exchange: RPF calculations 

Record no. Reference PAH Endpoint 

Data type: quantal or 

continuous BMR BMD 

Point 

estimate 

extra risk 

response 

Point 

estimate 

dose 

Dose 

units 

Converted 

dose 

Converted 

dose 

units RPF Comm

14620 Kochhar, 

1982 

BaP Aberrations Q 0.53 5 μg/mL 1 BMD doesn

selected BM

based on pe

 BaA 0.34 5 μg/mL 0.64 BMD doesn

selected BM

based on pe

14640 Krolewski 

et al., 1986 

BaP Sister 

chromatid exchanges 

C 0.79 5 μM 1.3 mg/L 1 

 CPcdP 0.29 5 μM 1.1 mg/L 0.41 No model fi

based on pe

response 

19690 Mane et al., 

1990 

BaP Sister 

chromatid exchanges 

C 2.7 1 μg/mL 1 

 BaA 0.4 1 μg/mL 0.15 21710 Tong et al., 

1981a 

BaP Sister 

chromatid exchanges 

C 92 1 × 10-4 M 25.2 mg/L 1 

 BaA 13 1 × 10-4 M 22.8 mg/L 0.16 No n provid

based on pe

response 

2 3 4 
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1 APPENDIX F. EXAMPLE CALCULATION OF RPF DETECTION LIMIT 

2 

3 

Table F-1. Example data for calculation of RPF detection limit 

Group Dose Number with tumors Number in group Incidence 

Extra risk 

responsea

Actual responses 

Control 0 2 30 0.067 NA 

Anthanthrene 0.25 2 29 0.069 NA 

Benzo[a]pyrene 0.25 24 30 0.800 0.786 

Theoretical statistically significant responseb

Anthanthrene 0.25 8 29 0.276 0.224 

aCalculated as described below in Step 1. 

bCalculated as described below in Step 2. 

Source: Hoffmann and Wynder (1966). 

4 

5 Step 1. Estimate the number of tumor-bearing animals that would represent a statistically6 significant response (one-sided p ≤ 0.05 using Fisher’s exact test) in the number of animals 

7 exposed to anthanthrene (29) given the observed control response (2/30). In this case, 

8 8/29 tumor-bearing animals (incidence of 0.276) would represent a statistically significant 

9 response to anthanthrene. 

10 

11 Step 2. Calculate the extra risk response associated with the theoretical statistically significant 

12 incidence for anthanthrene and the observed benzo[a]pyrene incidence as follows: 

13 

14 Extra risk response = P(d) – P(0)

15 [1-P(0)] 

16 

17 For the theoretical statistically significant response to anthanthrene, 

18 

19 Extra risk response = (0.276 – 0.067)/(1 – 0.067) = 0.224 

20 

21 Step 3. Calculate the RPF detection limit as the ratio of the slopes associated with extra risk 

22 response and the actual doses of anthanthrene and benzo[a]pyrene as follows: 

23 

24 RPF Detection Limit = (Theoretical anthanthrene extra risk response/dose anthanthrene)

25 (benzo[a]pyrene extra risk response/dose benzo[a]pyrene) 

26 

27 RPF Detection Limit = (0.224/0.25)/(0.786/0.25) = 0.28 

28 
G-1 DRAFT – DO NOT CITE OR QUOTE 

1 APPENDIX G: EVALUATION OF ALTERNATIVES FOR RANKING RPFs 

2 

3 

4 For many of the PAHs evaluated in this report, a number of datasets were available for 

5 use in calculating RPFs. The resulting RPFs are derived from tumor bioassays using different 

6 exposure routes, species, sexes, or tumor endpoints (incidence or multiplicity) and/or from a 

7 variety of different cancer-related endpoint assays. In addition to using different types of data, 

8 the various RPFs reflect studies of varying quality (different numbers of animals, follow-up time, 

9 single or multiple dose groups, response levels low or high on the dose-response curve, etc.). In 

10 order to derive a single final RPF for each individual PAH, the various results from different 

11 datasets must either be ranked/prioritized and/or combined. This appendix details the options 

12 that were considered for ranking RPFs. 

13 A series of options were considered for prioritizing RPFs for the purpose of selecting a 

14 single RPF for each PAH or exposure route. An a priori decision was made to consider tumor 

15 bioassay data to be preferable to cancer-related endpoint data because the tumor bioassay data 

16 are in whole animals and address the endpoint of interest for RPFs (tumorigenicity). Thus, 

17 options for ordering or combining tumor bioassays and for cancer-related endpoint data were 

18 considered separately; Section G.1 below discusses options considered for use of tumor bioassay 

19 RPFs and Section G.2 discusses options considered for use of cancer-related endpoint RPFs. 

20 

21 G.1. OPTIONS FOR RANKING TUMOR BIOASSAY RPFS 

22 Approaches considered for ordering tumor bioassay were: (1) ranking by exposure route, 

23 (2) ranking by target organ, and (3) preference for modeled data over point estimates. 

24 Ranking by exposure route. One option for ranking RPFs derived from tumor bioassay 

25 data would be to order the datasets by exposure routes that are considered most relevant to 

26 environmental exposure routes (oral, dermal, and inhalation). RPFs for many PAHs were 

27 calculated from dermal tumor bioassays. While dermal exposure to PAHs in the environment 

28 does occur, there is currently no dermal slope factor for benzo[a]pyrene on the IRIS database. If 

29 a dermal slope factor is derived for benzo[a]pyrene, then the RPFs derived from dermal tumor 

30 bioassays would be more suitable than those from other exposure routes for use in calculating 

31 cancer risk from dermal exposure to other PAHs. The available database for PAHs did not 

32 include any oral or inhalation studies that were suitable for RPF calculation; thus, route-to-route 

33 extrapolation is necessary to derive RPFs applicable to oral or inhalation exposures. 

34 Some earlier RPF approaches, primarily in the course of assessing risks from inhalation 

35 exposure to PAHs, have proposed hierarchies of bioassay types based on route of administration. 

36 Collins et al. (1998) proposed a hierarchy for PAH cancer potencies or PEFs for use in assessing 

37 air contaminants. The hierarchy for inhalation potencies or PEFs proposed by Collins et al. 

38 (1998) ordered the exposure routes as follows: intratracheal or intrapulmonary 

39 administration>oral administration>skin-painting studies> subcutaneous or intraperitoneal 
G-2 DRAFT – DO NOT CITE OR QUOTE 

1 administration. However, Collins et al. (1998) did not provide any empirical data supporting the 

2 ordering of these exposure routes, other than the intuitive preference for intratracheal or 

3 intrapulmonary administration as a surrogate for inhalation. In another review of data available 

4 for relative potency assessment for PAHs as air contaminants, Pufulete et al. (2004) suggested 

5 that intratracheal instillation of low doses of PAHs might be an appropriate surrogate exposure 

6 model for assessing relative potency of inhalation exposure. The basis for this suggestion was 

7 the authors’ observation that clearance of PAHs administered in solution via intratracheal 

8 instillation exhibited a biphasic pattern similar to that observed after inhalation exposure to 

9 benzo[a]pyrene bound to particulates. However, the authors acknowledged that the high 

10 concentrations of PAHs used in intratracheal and intrapulmonary instillation studies may lead to 

11 major differences in pharmacokinetics compared with inhalation exposure (Pufulete et al., 2004). 

12 Further, the authors expressed this suggestion as a path for future research, rather than as a 

13 means of examining available data on PAHs; no intratracheal instillation studies were identified 

14 in the search for studies from which to calculate RPFs for PAHs. Pufulete et al. (2004) did not 

15 provide any specific information on the relevance of intrapulmonary administration (a route used 

16 in several of the bioassays used to calculate RPFs) to inhalation exposure. 

17 To assess exposure-route differences in RPFs calculated in this review, a table comparing 

18 the average RPF for each PAH across exposure routes was prepared (Table G-1). The average 

19 values include RPFs calculated with both incidence and multiplicity data; each RPF is treated as 

20 an independent measure of relative potency in calculating the averages. Dermal studies are 

21 shown collectively as well as separated by study type (complete carcinogenesis or initiation 

22 only). Likewise, intraperitoneal studies are shown grouped as well as separated by target organ 

23 (lung and liver). In general, the table shows that RPFs calculated from lung implantation and 

24 dermal studies are similar, while RPFs calculated from intraperitoneal studies tend to be higher 

25 for most compounds. Among PAHs with RPF s derived from intraperitoneal and dermal data, 

26 5/7 showed higher RPF values from intraperitoneal data, compared with dermal data 

27 (benz[a]anthracene, benzo[b]fluoranthene, benzo[j]fluoranthene, cyclopenta[c,d]pyrene, 

28 dibenz[a,h]anthracene; Table G-1). However, intraperitoneal RPFs for chrysene (CH) and 

29 dibenzo[a,l]pyrene (DBalP) are similar to dermal RPFs for these compounds. 

30 
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1 

Table G-1. Average RPF value by exposure route 

Dermal 

Dermal 

complete 

Dermal 

initiation 

Intraperitoneal 

Intraperitoneal, 

target organ = 

lung 

Intraperitoneal, 

target 

organ = 

liver 

Lung 

implantation

PAH N Average N Average N Average N Average N Average N Average N Average 

AA 1 0.5 1 0.5 – – – – – – – – 1 0.2 

AC – – – – – – – – – – – – – – 

BaA 1 0.02 – – 1 0.02 3 0.3a 1 0.08 2 0.4 – – 

BbcAC 

(1,12-MBA) 1 0.05 – – 1 0.05 – – – – – – – – 

BbF 3 0.3 1 0.2 2 0.4 3b1

c

 1 0.4 1 2 1 0.1 

BeAC 4 0.7 – – 4 0.7 – – – – – – – – 

BghiP – – – – – – – – – – – – 1 0.009 

BjAC – – – – – – 1 60d 1 60 – – – – 

BjF 3 0.06 – – 3 0.06 5b0.6a 2 0.5 1 0.7 1 0.03 

BkF 2 0.02 – – 2 0.02 – – – – – – 1 0.03 

BlAC 4 4 – – 4 4 – – – – – – – – 

CH 7 0.1 – – 7 0.1 3 0.1a 1 0.1 2 0.2 1 0.04 

CPcdP 7 0.2 3 0.3 4 0.1 1 1

d

 1 1 – – – – 

CPdefC 2 0.3 – – 2 0.3 – – – – – – – – 

DBacA – – – – – – – – – – – – – – 

DBaeF 2 0.9 1 1 1 0.7 – – – – – – – – 

DBaeP 2 0.4 1 0.3 1 0.4 – – – – – – – – 

DBahA 2 1 – – 2 1 1 10d 1 10 – – 1 2 

DBahP 1 0.9 – – 1 0.9 – – – – – – – – 

DBaiP 2 0.6 1 0.7 1 0.5 – – – – – – – – 

DBalP 2 30 – – 2 30 1 30d 1 30 – – – – 

FA – – – – – – 10 0.08a 8 0.05 2 0.2 – – 

IP – – – – – – – – – – – – 1 0.07 

N23eP 1 0.3 – – 1 0.3 – – – – – – – – 

PH – – – – – – – – – – – – – – 

Pyr – – – – – – – – – – – – – – 

aNewborn mouse model. 

bNumber of intraperitoneal RPFs includes those calculated for combined lung and liver incidence; these 

are not included in number of RPFs with lung or liver tumors. 

c

Includes both newborn mouse and adult A/J mouse models. 

dAdult A/J mouse model. 

2 

3 One possible explanation for the higher intraperitoneal RPFs calculated from newborn 

4 mouse assays (footnoted “a” in the table) might be that the newborn mouse is more sensitive to 

5 the carcinogenic action of PAHs than an adolescent or adult mouse. Likewise, the adult A/J 

6 mouse is considered to be particularly sensitive to PAH lung tumorigenicity (Nesnow et al., 
G-4 DRAFT – DO NOT CITE OR QUOTE 

1 1995), which may result in higher RPFs with this model (in Table G-1, the intraperitoneal RPFs 

2 based on the A/J mouse model are footnoted “d”). There is little information to evaluate whether 

3 the newborn mouse is more or less sensitive than the adult A/J mouse model. Only one 

4 compound, benzo[b]fluoranthene (BbF), had RPFs calculated from both newborn mouse and 

5 adult A/J mouse models; the newborn mouse RPF was 2, while the A/J mouse RPF was 0.4. In 

6 summary, it is not clear whether the intraperitoneal RPFs are higher than dermal or lung 

7 implantation RPFs due to route-specific differences or animal model differences in susceptibility. 

8 Ranking by target tissue. An alternative approach to ranking tumor bioassay RPFs would 

9 be to prefer target tissue-specific RPFs (for example, to prefer RPFs derived from lung tumor 

10 data for inhalation RPFs). An analysis was conducted to assess whether RPFs calculated from 

11 lung tumor potency in intraperitoneal studies (both newborn mouse and adult A/J mouse models) 

12 were consistent with RPFs from lung implantation studies. Table G-1 shows RPFs calculated for 

13 lung tumors (separate from liver tumors also observed in some intraperitoneal studies) after 

14 intraperitoneal administration. Only four compounds (BbF, BjF, CH, and DBahA) had RPFs for 

15 both intraperitoneal and lung implantation studies; for each of these, the intraperitoneal lung 

16 tumor RPF exceeded the lung implantation RPF. No information assessing the concordance 

17 between lung tumor potency after intraperitoneal administration and inhalation cancer potency 

18 was identified in the literature.

19 Ranking by use of BMD. A third approach considered for ranking of tumor bioassay data 

20 was to prefer data amenable to BMD modeling (of either quantal or continuous data, depending 

21 on whether incidence or multiplicity was modeled) over an analysis of data based on point 

22 estimates. Table G-2 compares the average of RPFs for all bioassays with RPFs calculated using 

23 BMD modeling, and RPFs calculated using a point estimate approach. 

24 
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Table G-2. Comparisons among average tumor bioassay-based RPF values 

by data availability or calculation method 

All bioassays Multidose bioassays BMD model Point estimate 

N Average RPF N Average RPF N Average RPF N Average RPF 

AA 2 0.4 1 0.2 1 0.2 1 0.5 

AC – – – – – – – – 

BaA 4 0.2 – – – – 4 0.2 

BbcAC 1 0.05 1 0.05 – – 1 0.05 

BbF 7 0.6 5 0.3 3 0.2 4 0.9 

BeAC 4 0.7 4 0.7 2 0.8 2 0.7 

BghiP 1 0.009 1 0.009 1 0.009 – – 

BjAC 1 60 1 60 – – 1 60 

BjF 9 0.4 4 0.06 2 0.09 7 0.5 

BkF 3 0.02 3 0.02 2 0.03 1 0.02 

BlAC 4 4 4 4 2 5 2 3 

CH 11 0.1 5 0.1 3 0.1 8 0.1 

CPcdP 8 0.3 8 0.3 2 0.1 6 0.4 

CPdefC 2 0.3 – – 1 0.5 1 0.2 

DBacA – – – – – – – – 

DBaeF 2 0.9 1 1.1 1 1 1 0.7 

DBaeP 2 0.4 1 0.3 1 0.3 1 0.4 

DBahA 4 4 1 10 1 10 3 1 

DBahP 1 0.9 – – – – 1 0.9 

DBaiP 2 0.6 1 0.7 1 0.7 1 0.5 

DBalP 3 30 3 30 – – 3 30 

FA 10 0.08 8 0.08 5 0.08 5 0.07 

IP 1 0.07 1 0.07 1 0.07 – – 

N23eP 1 0.3 – – – – 1 0.3 

PH – – – – – – – – 

Pyr – – – – – – – – 

1 

2 While this ranking could be justified based on a general preference for multidose data and 

3 modeling to identify a point of departure, there are important limitations to this approach. First, 

4 RPFs based on BMD modeling may still use a point of departure high on the dose-response 

curve, if a single benzo[a]pyrene dose with an elevated response level (BMR)55 was used to 

6 calculate the RPF. In some cases, an RPF based on a point estimate approach from a point of 

7 departure lower on the dose-response curve may be a better predictor of relative potency at 

8 environmental exposure levels. Second, unless RPFs based on BMD modeling are available for 

9 all of the relevant exposure routes (dermal initiation and complete carcinogenicity, lung 

10 implantation, and intraperitoneal), there may be differences between the RPFs calculated from 

11 BMD modeling and those calculated using a point estimate approach that are unrelated to study 

12 quality (i.e., route, species, sex differences). Thus, ranking RPFs based on a preference for 

13 modeled data over point estimate data would neglect other sources of variability in the estimates 

14 (exposure route, species, sex, target organ, dosing intervals, etc.) 

 

5

The BMR selected for multidose PAH data for studies with a single BaP dose was the response level observed in 

the BaP dose group. 
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1 In summary, the analysis of options for ranking bioassay RPFs did not suggest a clear 

2 basis for selecting among the available data types. As a consequence, none of the available data 

3 types was considered preferable to any other; all bioassay RPFs were considered equally 

4 relevant. 

5 

6 G.2. RANKING NON-BIOASSAY DATA 

7 Two approaches to ranking non-bioassay study types were evaluated: a theoretical 

8 approach and an empirical approach.

9 Theoretical ranking of cancer-related endpoint data. To identify whether a theoretical 

10 basis for ranking was available, a limited literature search was conducted in PubMed to identify 

11 publications that addressed the relationship between various genotoxicity endpoints and 

12 carcinogenicity. The search was intended to identify recent papers that described a quantitative 

13 correlation between particular cancer-related endpoints and cancer potency. Several papers (e.g., 

14 Matthews et al., 2006a, b) analyzed the concordance between genotoxicity tests and 

15 carcinogenicity (i.e., whether positive genotoxicity findings predicted positive carcinogenicity 

16 findings) but did not assess whether potency measured in genotoxicity studies correlated with 

17 cancer potency. Three publications (Sanner and Dybing, 2005; Ross et al., 1995; Travis et al., 

18 1990) provided quantitative associations between cancer potency and genotoxic potency. There 

19 were no publications that related relative cancer potency to relative genotoxic potency for any 

20 classes of compounds; this would be the most relevant comparison for use in ranking. 

21 Ross et al. (1995) provided evidence that DNA adduct formation expressed as TIDAL 

22 values was correlated with the numbers of lung adenomas in adult A/J mice treated with PAHs 

23 by intraperitoneal injection (TIDAL values were also strongly correlated with dose). The 

24 correlations were demonstrated for five PAHs: benzo[a]pyrene, benzo[b]fluoranthene, 

25 dibenz[a,h]anthracene, 5-methylcholanthrene, and CPP. This paper demonstrates a quantitative 

26 relationship between TIDAL values and tumorigenicity in the compound class of interest 

27 (PAHs). 

28 Examining data from 42 substances, Sanner and Dybing (2005) showed a linear 

relationship (slope = 1.05 ± 0.12, r229 = 0.67) between the lowest effective dose (LED) producing 

30 genotoxicity in vivo after oral or inhalation exposure and the chronic daily dose estimated to 

31 result in a 25% increase (over controls) in tumor formation (TD25). Genotoxicity endpoints 

32 included in the analysis were micronuclei, sister chromatid exchanges, chromosomal aberrations, 

33 DNA adducts, DNA strand breaks, and the comet assay for DNA breaks. The chemical 

34 compounds included in the analysis included a large variety of compounds (VOCs, chlorinated 

35 compounds, dioxin, etc.); only one PAH (naphthalene) was included in the database. Analyzing 

36 specific endpoints separately, the authors reported the correlations shown in Table G-3. 

37 
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Table G-3. Correlation between LED and TD25 by endpoint

Endpoint 

Number of observations 

(compounds) Slope 

Correlation coefficient 

(r2) 

Chromosomal aberrations 4 1.29 ± 0.25a 0.93 

DNA breaks 9 1.02 ± 0.17a 0.84 

DNA adduct 11 1.44 ± 0.36a 0.64 

Micronucleus 8 0.63 ± 0.21a 0.61 

Sister chromatid exchange 2 0.98 – 

a

Slope significantly different from zero. 

Source: Sanner and Dybing (2005). 

1 

2 The slopes and correlation coefficients shown in Table G-3 suggest that LEDs for 

3 chromosomal aberrations and DNA breaks are reasonably good predictors of cancer TD25, and 

4 that DNA adducts and micronuclei are also correlated with potency. 

5 Travis et al. (1990) examined correlations between cancer potency estimates and 

6 mutagenicity or toxicity data from the Registry of Toxic Effects of Chemical Substances 

7 (RTECS). Using data from 146 compounds, the authors demonstrated correlations between 

8 tumor potency and mutagenicity or toxic potency. Correlations between toxicity and 

9 carcinogenicity were not considered here, as they are not pertinent to the ranking of genotoxicity 

10 data for RPF selection. Correlations between mutagenicity and cancer potency are shown in 

11 Table G-4. 

12 

Table G-4. Correlation between tumor potency (log 1/TD50) 

and mutagenic potency

Endpoint 

Number of observations 

(compounds) 

Correlation 

coefficient (r2) 

Standard 

error 

Ames test 82 0.37 0.93 

All mutation tests 112 0.69 0.90 

All mutation tests: lung carcinogensa 17 0.61 0.74 

All mutation tests: liver carcinogensa 40 0.43 0.94 

aDue to the magnitude of the tumor potency for tetrachlorodibenzo-p-dioxin (TCDD), it had a strong influence on 

the correlation; the authors conducted analyses without dioxin, and these are the results shown here. 

Source: Travis et al. (1990). 

13 

14 The available published studies did not suggest a clear basis for ordering RPFs calculated 

15 from data on different cancer-related endpoints. Based on the information provided in the three 

16 published analyses relating cancer potency with genotoxic potency, one may conclude that 

17 TIDAL values, in vivo measures of clastogenicity, DNA adducts, and DNA damage, and in vitro 
G-8 DRAFT – DO NOT CITE OR QUOTE 

1 measures of mutagenicity are correlated with some measure of cancer potency. While it could be 

2 argued that a stronger basis for the correlation exists for TIDAL values (based on Ross et al., 

3 1995), since the data for this endpoint were collected for the compound class of interest (PAHs), 

4 there is no information to determine whether TIDAL values are better predictors of cancer 

5 potency in PAHs than the other available endpoints, because other endpoints were not examined 

6 for PAHs as a class. In addition, the measures of cancer and genotoxicity potency used in the 

7 three papers were different, making comparisons among the three difficult. Based only on the 

8 analysis conducted by Sanner and Dybing (2005), one might use the slopes and/or correlations 

9 (Table 3) to rank in vivo genotoxicity endpoints, but it is not clear where TIDAL values would 

10 fit, as these were not analyzed separately. TIDAL values could be categorized with other DNA 

11 adduct measures assessed by Sanner and Dybing (2005); however, Ross et al. (1995) 

12 demonstrated that adduct levels measured at a single time point were uncertain predictors of 

13 potency compared with TIDAL values. 

14 Empirical ranking of cancer-related endpoint data. In view of the fact that no published 

15 studies comparing relative genotoxic potency to relative cancer potency were available, and that 

16 the present work created a large database of RPFs for multiple endpoints, an empirical approach 

17 to assigning ranks was also explored. The database of PAH RPFs was analyzed to determine 

18 whether any individual cancer-related endpoint was more closely correlated with RPFs based on 

19 tumor bioassay data. The premise behind this analysis is that RPFs based on bioassay data 

20 represent the best available information, and that the genotoxicity endpoints that best predict 

21 bioassay RPFs should be preferred over those that show little relationship to tumor bioassay 

22 RPFs. The semiquantitative analysis was, of necessity, restricted to those PAHs for which at 

23 least one RPF based on bioassay data was available.

24 For each of the 22 PAHs with nonzero RPFs based on bioassay data, the average bioassay 

25 RPF was compared with the average RPF for several endpoints that the literature review 

26 suggested could be correlated with cancer potency (in vivo DNA adducts, in vivo micronuclei 

27 and sister chromatid exchanges together, and in vitro mutagenicity). TIDAL values were not 

28 analyzed separately from other measures of DNA adducts because there were only four PAHs 

29 with both TIDAL and bioassay RPFs; similarly, micronuclei and sister chromatid exchange 

30 endpoints were grouped to increase the number of observations in the regression. In addition to 

31 analyzing these endpoints, an analysis of several endpoints grouped across class (e.g., all in vivo 

32 non-bioassay endpoints, all in vitro endpoints, and all non-bioassay endpoints) was performed. 

33 Linear regression was performed on the log-transformed average RPF values to assess the 

34 predictive power of each endpoint or grouping, and to assess whether there was a quantitative 

35 basis for ordering them. 

36 Table G-5 shows the results of regression analyses assessing how well the average RPFs 

37 for several endpoints correlated with average bioassay RPFs. The table shows that neither in 

38 vivo clastogenicity RPFs (micronuclei and sister chromatid exchanges) nor in vitro mutagenicity 
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1 RPFs were significantly correlated with bioassay RPFs for the dataset examined here. Among 

2 those showing a significant (p < 0.05) linear relationship, in vivo DNA adducts provided the best 

correlation (r2 = 0.64), followed by all in vivo non-bioassay endpoints (r23 = 0.54), all nonbioassay endpoints (r2

 = 0.43), and all in vitro non-bioassay endpoints (r

2

4 = 0.39). Although in 

5 vivo DNA adducts provided the strongest correlation, the slope for this regression was 1.25, 

6 indicating that RPFs for in vivo DNA adducts systematically underpredicted bioassay RPFs. 

7 Figure G-1 demonstrates this underprediction; as the figure shows, most of the average RPF 

8 values are to the left of the 1:1 correspondence line. The slopes for both in vivo non-bioassays 

9 and all non-bioassays are much closer to 1.0. Plots showing the average RPF comparisons for all 

10 in vivo non-bioassays, all non-bioassays, and all in vitro non-bioassays are shown in Figures G-2 

11 through G-4. These plots suggest that in vivo non-bioassay RPFs tend to underpredict bioassay 

12 RPFs, while all in vitro non-bioassays tend toward overprediction. 

13 

Table G-5. Results of simple linear regression of log–transformed average 

genotoxicity RPF vs. log average tumor bioassay RPF

Genotoxicity endpoint R

2

 Slope p-Value n 

All in vivo DNA adducts 0.64 1.24 <0.01 9 

All in vivo non-bioassays 0.54 1.05 0.016 10 

All non-bioassay endpoints (in vitro and in vivo) 0.43 1.03 <0.01 19 

All in vitro non-bioassays 0.39 0.91 <0.01 19 

All in vivo micronuclei and sister chromatid exchanges 0.58 0.83 >0.05 (NS) 6 

All in vitro mutagenicity 0.047 0.39 >0.05 (NS) 17 

14 
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0.001

0.01

0.1

1

10

100

0.001 0.01 0.1 1 10 100

Average in vivo DNA adduct RPF

Average bioassay RPF

1 

2 Figure G-1. Average bioassay RPF vs. average in vivo DNA adduct RPF. 

0.001

0.01

0.1

1

10

100

0.001 0.01 0.1 1 10 100

Average in vivo nonbioassay RPF

Average bioassay RPF

3 

4 Figure G-2. Average bioassay RPF vs. average in vivo nonbioassay RPF. 

5 
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0.001

0.01

0.1

1

10

100

0.001 0.01 0.1 1 10 100

Average non-bioassay RPF

Average bioassay RPF

1 

2 Figure G-3. Average bioassay RPF vs. average nonbioassay RPF. 

0.001

0.01

0.1

1

10

100

0.001 0.01 0.1 1 10 100

Average in vitro nonbioassay RPF

Average bioassay RPF

3 

4 Figure G-4. Average bioassay RPF vs. average in vitro non-bioassay RPF. 

5 

6 Based on the results of the linear regression analyses comparing PAH RPFs calculated 

7 for genotoxicity endpoints and RPFs calculated for bioassays (Table G-5), an argument could be 
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1 made for the following ranking: (1) bioassays, (2) in vivo non-bioassays, and (3) in vitro non2 bioassays. However, the improvement in correlation that is achieved with subdividing all nonbioassays into in vivo and in vitro endpoints is small (r2

3 improves from 0.43 for all nonbioassays 

4 to 0.54 for in vivo non-bioassays), and the plot for in vivo nonbioassay shows that this grouping 

5 exhibits a slight tendency to underpredict bioassay RPFs. 

6 In summary, as with the findings for tumor bioassay data, the analysis of options for 

7 ranking cancer-related endpoint RPFs did not suggest any clear theoretical or empirical basis for 

8 prioritizing the available data for the purpose of selecting RPFs. Thus, for PAHs without any 

9 tumor bioassay RPFs but with adequate information to suggest potential carcinogenicity, the 

10 cancer-related endpoint data were combined to calculate a final RPF as described in Section 7. 

11 

12