Testing of Certain High Production Volume Chemicals; 

Third Group of Chemicals

Final Rule

RESPONSE TO PUBLIC COMMENTS

Prepared by the Chemical Information and Testing Branch,

Office of Pollution Prevention and Toxics,

Office of Chemical Safety and Pollution Prevention, 

U.S. Environmental Protection Agency

September 12, 2011
2

Table of Contents

Unit Title Page

A. Introduction 4

B. General Comments and the Use of Existing Information 5 

C. Comments on Proposed Test Methods 5

 

 1. Acute toxicity test method 5

2. Aquatic toxicity test method 7

3. Genetic toxicity test method 9

4. Reproductive and developmental toxicity screening 9

 

D. Animal Welfare Considerations 11

 

E. Chemical Specific Comments 12

 

 1. Benzene, 1,2-dimethyl-3-nitro- (CASRN 83-41-0) 12

 

 2. 3-Pentanone (CASRN 96-22-0) 14 

 

 3. Benzene, 1-chloro-4-(trifluoromethyl)- (CASRN 98-56-6) 15

 

 4. Benzenesulfonic acid, 3-nitro, sodium salt (1:1) (CASRN 127-68-4) 17

 5. 1-Tetracosanol (CASRN 506-51-4); 1-Hexacosanol (CASRN 506-52-5) 19

 

 6. 2-Propenoic acid, 2-carboxyethyl ester (B-CEA) (CASRN 24615-84-7 25

 

 7. Benzenesulfonic acid, dimethyl- (CASRN 25321-41-9) 26

 

 8. Methanesulfonamide, N-[2-[(4-amino-3-methylphenyl)ethylamino]ethyl]-, 

sulfate (2:3) (CASRN 25646-71-3) 31 

 9. 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) 

(CASRN 52556-42-0). 34

 

 
3

 10. Coal/Coal Tar Chemical Substances: 35

Solvent naphtha (coal) (CASRN 65996-79-4);

Tar oils, coal (CASRN 65996-82-9);

Tar, coal, high-temperature (CASRN 65996-89-6);

Distillates (coal tar) (CASRN 65996-92-1);

Pitch, coal tar-petroleum (CASRN 68187-57-5);

Extract residues (coal), tar oil alk., naphthalene distn. 

 residues (CASRN 73665-18-6)

 11. 1,4-Benzenedicarboxylic acid, 1,4-dimethyl ester, manuf. of, 

by-products from (CASRN 68988-22-7) 48

 

 12. 1-Decene, sulfurized (CASRN 72162-15-3) 53

 

Bibliography 56

 
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A. Introduction

This document contains a summary of the comments received by EPA in response to the 

TSCA section 4 proposed test rule entitled “Testing of Certain High Production Volume 

Chemicals, Third Group of Chemicals” (75 FR 8575, February 25, 2010) along with EPA’s 

responses to those comments. The proposed rule and comments are available in EPA’s public 

docket and e-docket for the final test rule under Docket No. EPA–HQ–OPPT–2009–0112. EPA 

received comments from industry, including: 

• American Chemistry Council (ACC)

• The American Coke and Coal Chemicals Institute (ACCCI)

• BASF

• Chlorinated Paraffins Industry Association (CPIA)

• Eastman Chemical Company (Eastman)

• Greenwich Chemical Consulting, Inc. (GCC)

• International Imaging Industry Association (I3A)

• INVISTA

• IsleChem, LLC (IsleChem)

• Rhodia, Inc. (Rhodia)

• Sasol North America (SASOL)

• GE Water and Process Technologies (GE)

• Special Materials Company (SMC)

Greenwich Chemical Consulting, Inc. (GCC) submitted comments on behalf of Brenntag 

North America, Inc.; Kowa American Corporation; Miami Chemical, Inc.; and Univer USA, 

Inc. (“The Companies”).

EPA also received comments from a number of Animal Welfare Organizations (AWOs). 

AWOs that submitted comments under the letterhead of People for the Ethical Treatment of 

Animals (PETA) included PETA, the Physicians Committee for Responsible Medicine 

(PCRM), the Alternatives Research Development Foundation, and the American Anti-
5

Vivisection Society. EPA also received a comment from a private citizen concerned with 

companies using animals to test their commercial products and advocated other (non-animal) 

tests for this purpose.

B. General Comments and the Use of Existing Information

Comment. AWOs commented that, in cases where chemical properties and environmental fate 

data are lacking, no toxicity tests should be conducted until after such basic testing is 

completed, as certain chemical/fate properties may preclude the need to conduct toxicity tests.

Response. While it is true that physical chemical property and fate data can be useful in 

determining how specific toxicity tests are performed, few chemical and fate properties negate 

the requirement for testing altogether. Those physical properties that are most related to 

bioavailability, the water solubility and octanol-water partition coefficient, are readily 

estimated using chemistry modeling computer programs, should these data be necessary for 

experimental design. With the exception of intrinsic chemical reactivity, knowledge of a 

chemical’s environmental fate is generally not necessary prior to assessing its toxicity. It is 

agreed that highly reactive chemicals that undergo instantaneous destruction upon exposure to 

the environment should not be tested for toxicity. However, reactivity of this nature is generally 

well known to those who manufacture, transport, use or otherwise handle the chemical in 

question. In addition, the reactivity of the parent does not necessarily preclude testing of the 

degradation products. Therefore, unless there is a question regarding the reactivity of the 

chemical, there is no reason to withhold toxicity testing until all of the chemical/fate property 

testing has been conducted.

C. Comments on Proposed Test Methods

1. Acute toxicity test method.

Comment. AWOS commented that the acute toxicity testing requirement may not be necessary. 

According to AWOs, evidence indicates that more than 85% of industrial chemicals are non-
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toxic (Kinser-Ovaskainen et al., 2008) and efforts to validate the in vitro 3T3 NRU cytotox 

assay (ICCVAM, 2006) also indicate that this assay correctly discriminates non-toxic 

chemicals from toxic ones. AWOs suggest that EPA wait for the final results of this validation 

before requiring in vivo (acute mammalian) toxicity testing, which is specified in the test rule.

AWOs stated that a coalition of European pharmaceutical companies determined that 

regulatory decisions were almost never predicated on the results of acute toxicity tests, which 

prompted the International Council on Harmonization and its regional pharmaceutical 

regulators to propose removing the requirement for an acute oral toxicity testing from the 

testing guidelines (Robinson et al., 2008; European Medicines Agency, 2008) and obtain 

necessary information on acute toxicity from available repeated dose studies. According to 

AWOs, the European Center for the Validation of Alternative Methods (ECVAM) used the 

approach of the International Council on Harmonization and correctly classified a majority of 

chemicals (Kinsner-Ovaskainen et al., 2008). Additionally, AWOs state that ECVAM is 

currently conducting a definitive validation of this hypothesis with thousands of chemicals. 

AWOs also suggested that an emerging method to estimate toxicity for exposures via the 

“inhalation route” (Mackay et al., 2008; Veith et al., 2008) should also be pursued. A similar 

comment was made by AWOs in response to the TSCA section 4 proposed test rule entitled 

“Testing of Certain High Production Volume Chemicals, Second Group of Chemicals” (73 FR 

43314, July 24, 2008).

Response. While EPA generally agrees that, for a variety of reasons, test methods which 

minimize the use of live animals are desirable, a new in vitro method, such as one that would 

estimate toxicity for exposures via the “inhalation route,” must be validated before in vivo

testing can be partially or completely eliminated. In the case of the 3T3 NRU cytotoxicity 

assay, EPA agrees with the conclusion of a 2006 ICCVAM Test Method Evaluation Report 

(ICCVAM, 2006); namely, that data from the 3T3 NRU cytotoxicity assay may be used in a 

weight-of-evidence approach, where appropriate, for determining starting doses for in vivo

acute oral systemic toxicity studies (EPA 2008b). EPA acknowledges that this use of this 

approach may result in the reduction of animals used in acute oral toxicity testing. As discussed 

below, EPA also agrees with ICCVAM’s conclusion that these in vitro test methods are not 
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sufficiently validated to replace animals for regulatory labeling and hazard classification 

purposes (EPA 2008b). 

Although EPA agrees that the 3T3 NRU assay can be used in a weight-of-evidence 

approach, it should be acknowledged that the conclusions of the ICCVAM report, as well as an 

OECD guidance document on the use of the 3T3 NRU cytotox assay were based on validation 

studies conducted on only 72 chemicals. EPA agrees that an evaluation of thousands of 

chemicals using the 3T3 NRU cytotoxicity assay would be more desirable in determining the 

true validity of the assay. However, the results of the validation by ECVAM using thousands of 

chemicals are not available for EPA to evaluate as of this writing. It should also be noted that 

the 3T3 NRU cytotox assay is for the purpose of estimating starting doses -- not as a complete 

replacement of in vivo testing -- for acute oral systemic toxicity tests. Although the 3T3 cytotox 

assay is not currently accepted as a substitute for in vivo testing, EPA allows the use of in vitro

testing and similar approaches, such as Structure-Activity Relationship (SAR) models, to 

estimate the likelihood of toxicity of chemicals and thereby reduce the number of animals 

necessary to determine toxicity. 

2. Aquatic toxicity test method.

Comment. AWOs stated that in vivo aquatic toxicity testing is unnecessary considering the 

availability of suitable in silico and in vitro test methods such as ECOSAR and the DarT test.

Response. At the current time, the DarT test is not considered to be adequately validated to use 

as a replacement for acute fish toxicity testing (see response to next comment below). 

ECOSAR modeling, while useful in estimating toxicity, is not considered to be an acceptable 

method for the definitive determination of toxicity of chemicals for regulatory purposes. 

Comment. AWOs commented that the DarT test, using fertilized zebrafish eggs as a surrogate 

for living fish, has been nominated for development into an OECD test guideline and should be 

used as a replacement for (acute) fish toxicity testing. If fish acute toxicity testing is still 

required, a threshold (step-down) test (e.g., Jerama et al., 2005) should be used, with an 
8

additional refinement also possible to terminate testing after 24 hours of exposure in some 

cases (Hutchinson et al., 2003). AWOs stated that the Fish Threshold Approach (FTA) is a 

modification of the step-down approach and that an OECD Guidance Document describing this 

approach is in the process of being finalized (OECD, 2010).

Response. EPA notes that the DarT test has received attention as a potential in vitro

replacement for the current acute fish toxicity testing (e.g., Brannen et al., 2009). However, this 

test method is still being refined and undergoing validation. Until the DarT test has been

adequately validated, the Agency will continue to recommend approved guidelines, rather than 

methods that are still under development, which may in fact not be adequately predictive. The 

acute fish toxicity test conducted in vivo will continue to be required. 

Acute aquatic toxicity testing determines a 96-hour LC50; i.e., the concentration at which 

50% mortality is observed after 96 hours of exposure to a chemical. The Hutchinson et al. 

(2003) method appears to suggest that testing be terminated at 24 hours of exposure when 

lethality and/or serious morbidity are observed in two out of five fish (as described by AWOs). 

The authors state that this method could be used to interpolate an LC50 value. While this may be 

useful for estimation purposes, this method of extrapolation is not acceptable to 

EPA for determining an LC50. 

Comment. AWOs suggested that toxicity testing should be conducted first on aquatic plants and 

invertebrates, as toxicity to these organisms (fish food supply) might obviate the need for fish 

toxicity testing.

Response. EPA disagrees. Aquatic plants and invertebrates are included with fish in the 

evaluation of aquatic toxicity in order to determine the toxicity of a chemical to three different 

taxonomic levels of the aquatic ecosystem. In addition, it is possible that a chemical substance 

may be much more toxic to fish than to aquatic plants or invertebrates. Therefore, it is essential 

that all three tests be performed to adequately assess the risk to aquatic organisms. 
9

3. Genetic toxicity test method.

Comment. AWOs suggested that the genetic toxicity sections of the proposed rule should be 

amended to mandate (rather than encourage) the use of in vitro genetic toxicity testing; with the 

requirement that any company proposing to use an in vivo method first provide a compelling 

justification for an exemption from the in vitro test methodology, with opportunity for public 

comment. AWOs stated that in response to comments on the first proposed test rule, EPA 

anticipated that test sponsors would perform less expensive in vitro tests unless there were 

sound scientific reasons not to do so. However, AWOs indicated that retrospective analysis of 

all test plans submitted to the HPV Challenge Program indicated that only 21% of in vivo

chromosomal aberration tests originally proposed were subsequently replaced by in vitro tests

in revised test plans.

Response. In the proposed rule, and in its comments to test sponsors, EPA has made it clear that 

in vitro testing is preferred for collecting screening data for genetic toxicity endpoints. In the 

HPV Challenge Program and in the proposed rule, EPA has asked sponsors who submit test 

plans for in vivo genetic toxicity testing to include justification for the in vivo method instead of 

the in vitro method. EPA does not mandate that genetic toxicity testing be conducted in vitro in 

the HPV program because, as stated in the proposed rule, chemical properties may preclude use 

of the in vitro method. Also, extant in vivo genetic toxicity data for an HPV chemical or a 

supporting HPV chemical compound may be adequate to satisfy the genetic toxicity endpoints 

so that further testing is not required. In cases where an HPV chemical requires genetic 

mutation or chromosomal aberrations testing, EPA permits the use of in vivo testing, but only 

after those providing the testing present a strong justification for in vivo testing in lieu of in 

vitro testing.

4. Reproductive and developmental toxicity screening.

Comment. AWOs commented that EPA should amend the proposed rule to require companies 

to use the combined repeated-dose toxicity study with the reproduction/developmental toxicity 

screening test unless, before testing, a compelling justification for an exemption from the 
10

combined protocol is provided, with opportunity for public comment available. AWOs stated 

that in response to comments on the first proposed test rule, EPA anticipated that test sponsors 

would perform less expensive combined tests unless there were sound scientific reasons not to 

do so. However, AWOs indicated that retrospective analysis of all test plans submitted to the 

HPV Challenge Program showed that only 35% of individual repeated-dose, reproductive and 

developmental toxicity tests originally proposed were subsequently replaced by proposals for 

combined repeated-dose reproductive/developmental toxicity screening tests.

Response. EPA does, in fact, recommend use of the Combined Repeated Dose Toxicity Study 

with the Reproduction/Developmental Toxicity Screening Test (40 CFR 799.9365). However, 

there may be valid reasons to test a particular chemical using both 40 CFR 799.9355 

(Reproduction/Developmental Toxicity Screening Test) and 40 CFR 799.9305 (Repeated Dose 

28-Day Oral Toxicity Study in Rodents) to fill the Mammalian Toxicity - Repeated 

Dose/Reproduction/Developmental data needs, and is requiring in the final test rule that anyone 

who uses the combination of 40 CFR 799.9355 and 40 CFR 799.9305 in place of 40 CFR 

799.9365 submits to EPA a rationale for conducting these alternate tests.

Comment. AWOs stated that an embryonic stem cell test (EST) has been validated by the 

ECVAM as a test for embryotoxicity (Buesen et al., 2009; Festag et al., 2007a, 2007b; 

Genschow et al., 2002; Spielmann, 2009; van Dartel et al., 2010). AWOs suggested that this 

test is ideally suited for the HPV Challenge Program whereby chemicals testing positive for 

embryotoxicity could be classified as probable developmental toxicants without further testing.

Response. EPA does not agree. In a recent article on the embryonic stem cell test (EST; 

Buensen et al. 2009), the authors and developers of the original EST assay acknowledge that 

the version of the test that was validated by the ECVAM has since been shown to have some 

significant drawbacks. The main drawbacks, from EPA’s perspective, are the test’s reliance on 

a morphological endpoint (contracting cardiomyocytes) and the training and validation required 

to ensure that personnel can reliably obtain repeat assessments for this endpoint. The 

measurement for this endpoint is subjective rather than objective. In an attempt to address these 

issues, as well as the issue of time and expense of testing, Buensen et al. (2009) have 
11

established a new version of the EST, which they refer to as molecular fluorescence activated 

cell sorting (FACS)-EST. However, the authors acknowledge that further studies which 

incorporate a much wider selection of test compounds are necessary to assess the capabilities 

and limitations of the new technique.

D. Animal Welfare Considerations

Comment. AWOs expressed concern that EPA is still proposing extensive animal testing of 

HPV chemicals and that these proposals continue to ignore existing information and basic 

animal welfare principles that EPA enumerated in its October 14, 1999 letter to program 

participants. AWOs stated that experience gained during the review of hundreds of test plans 

should be applied to reduce the need for additional animal testing. This experience was 

summarized in comments submitted by the Physicians Committee for Responsible Medicine 

(PCRM) to EPA on June 4, 2008 regarding the Chemicals Assessment and Management 

Program (ChAMP). The comments by PCRM were included as an attachment to AWO 

comments on the proposed test rule.

Response. EPA agrees with AWOs that experience gained during the review of hundreds of test 

plans should be applied to reduce the need for additional animal sacrifices testing. The testing 

in this rule was required only after extensive efforts by industry, public commenters and EPA 

scientists to determine the availability of extant data to satisfy the required aquatic and human 

health endpoints for the test rule chemicals, the availability of supporting chemical data that 

would cover the needed endpoints in lieu of the need to collect further data, the use of available 

in vitro data where acceptable, as well as modeled data and weight-of-evidence approaches 

where appropriate. All available and currently acceptable approaches combined were used to 

eliminate as much additional animal testing as possible. After exhausting all other available 

means available to satisfy the required endpoints, the Agency determined that tests involving

the use of animals for these 16 chemicals were required to provide the data needed to guide risk 

assessments and risk management actions that would protect the environment and human 

health. While additional test rules for HPV chemicals may be promulgated by EPA, similar 

approaches as outlined above will be taken to minimize the use of live animals in testing.
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Once the required data are obtained on these chemicals, they will add to our 

compendium of knowledge on the toxicity of chemicals. EPA expects that this additional 

knowledge will aid in reducing the need for further animal testing on comparable chemicals in 

the future.

Comment. AWOs urged OPPT to implement an approach outlined in the National Research 

Council’s report, “Toxicity Testing in the Twenty-first Century: a Vision and a Strategy” 

(NRC, 2007) in which a recommendation was made to shift from whole animal tests that assess 

apical endpoints toward more human-relevant in silico and in vitro methods that assess toxicity 

at the molecular level.

Response. EPA, the National Toxicology Program (NTP) and the National Institutes of Health 

Chemical Genomics Center (NCGC) have formed a partnership known as Tox21, dedicated to 

the advancement of toxicity testing. Tox21 has responded to the NRC report with the 

publication in March 2009 of The U.S. Environmental Protection Agency’s “Strategic Plan for 

Evaluating the Toxicity of Chemicals” (http://www.epa.gov/osa/spc/toxicitytesting/). In this 

plan, EPA states its commitment to the use of new scientific tools in computational, 

information and molecular sciences to strengthen toxicity testing and risk assessment 

approaches. The change from using studies in animals to a reliance on results from biochemical

and cell-based assays for regulatory decisions is a process requiring sufficient time to 

determine that predictions made by these new methods are adequate and reliable. 

E. Chemical Specific Comments:

1. Benzene, 1,2-dimethyl-3-nitro- (3-NOX) (CASRN 83-41-0).

Comment. BASF, the only producer of this chemical substance, indicates in their comment that 

3-NOX is no longer a HPV chemical. Data from the most recent IUR cycles (2002 and 2006) 

show that the manufacture/import of this substance is well below the 1 million lb threshold 

established for HPV status. BASF noted that its IUR-reported production volume of 3-NOX 
13

has been less than 100,000 pounds since 2002, and stated that the company has no plans to 

exceed 100,000 pounds in the future. BASF also submitted information indicating that there is 

some additional production of 3-NOX. However, this additional production of 3-NOX is as a 

byproduct produced during the production of another registered chemical and this byproduct is 

incinerated and managed according to a RCRA permit. Therefore, the chemical is not a high 

production volume chemical and is not subject to the rule. 

Response: BASF is the only producer of this chemical substance and EPA finds the 

additional information submitted by the company supportive of its claims. Therefore, EPA

agrees 3-NOX appears to no longer meet the production volume or environmental release Bfinding thresholds and is not including this chemical in this final test rule. However, EPA may 

reevaluate this decision based on IUR data to be reported later in 2011. In the event IUR or 

other data provides new or additional support for the TSCA section 4(a)(1)(B)(i) finding for 3-

NOX, EPA will take appropriate steps to proceed with a test rule for this chemical substance.

Comment. AWOs noted that the proposed test rule requires mammalian acute toxicity and 

combined repeated-dose reproduction/developmental toxicity screening tests for 3-NOX. 

AWOs found an abstract from a 1994 German toxicology assessment (Anonymous, 1994) in 

the TOXLINE database that summarized results of acute and repeated-dose oral toxicity of 3-

NOX in rats. AWOs stated that EPA should review the information in this assessment 

regarding acute and repeated-dose toxicity and determine whether reproductive organs were 

examined in the repeated dose toxicity study.

Response: EPA retrieved the 1994 German toxicology assessment for 3-NOX (BG Chemie, 

1994) that was mentioned by the AWOs. Although the information regarding acute and 

repeated-dose (28-day) oral toxicity studies in rats were cited to unpublished studies from 

chemical companies in Germany, the BG Chemie (1994) summaries of these studies provided 

sufficient information for EPA to consider the studies to adequately characterize the acute and 

repeated-dose hazards of oral exposure to 3-NOX. 
14

Comment. AWOs stated that data for a “similar” chemical (1,2-dimethyl-4-nitrobenzene 

[CASRN 99-51-4]) were submitted to the HPV Program and that EPA considered the 

submitted data adequate for assessing health effects endpoints for 1,2-dimethyl-4-nitrobenzene. 

AWOs noted that data for several analogues, including 2,4-dimethyl-1-nitrobenzene (CASRN 

89-87-2) and 4-nitrotoluene (CASRN 99-99-0), were used in the HPV submission for 1,2-

dimethyl-4-nitrobenzene to complete the SIDS dataset and that EPA considered results of 

existing reproductive and developmental toxicity data for 4-nitrotoluene to be adequate. AWOs 

urged EPA to consider this additional information in its assessment of 3-NOX. AWOs consider 

it likely that the weight of the additional evidence is sufficient to eliminate the toxicity testing 

requirements for this chemical. The toxicity testing requirements in the proposed test rule for 

benzene, 1,2-dimethyl-3-nitro- include acute mammalian toxicity, in vitro or in vivo

chromosomal damage, and a combined repeated-dose reproductive/developmental screening 

test.

Response. In its assessment of the HPV submission for 1,2-dimethyl-4-nitrobenzene, EPA 

noted physical/chemical similarities between the sponsored chemical and analogues that 

included 2,4-dimethyl-1-nitrobenzene and 4 nitrotoluene. The determination that submitted 

information for the sponsored chemical and the analogs was adequate for assessing health 

effects was based in part on comparative information for acute toxicity and genotoxicity 

endpoints between sponsored and analogue chemicals. However, in the absence of information 

regarding acute toxicity, repeated-dose toxicity, reproductive/developmental toxicity, or 

genotoxicity of benzene, 1,2-dimethyl-3-nitro-, comparisons with data from analogue 

chemicals are not possible. Therefore, if 3-NOX still met the finding of “substantial 

production,” EPA would still require these mammalian toxicity testing requirements listed in 

the proposed test rule.

2. 3-Pentanone (CASRN 96-22-0).

Comment. AWOs stated that the proposed test rule requires mammalian acute toxicity and 

combined repeated-dose reproduction/developmental toxicity screening tests for 3-pentanone. 

AWOs noted that a search of fragrance raw materials monographs identified a reproductive 
15

toxicity study of 3-pentanone in mice (Carlson et al., 1975). AWOs further noted that a 

structurally similar chemical, 2-pentanone (CASRN 99-51-4) was sponsored in the HPV 

Challenge program and that all health effects endpoints were fulfilled for this chemical. AWOs 

urged EPA to consider the existing data for 2-pentanone, and other short chain aliphatic 

ketones such as methyl ethyl ketone (CASRN 78-93-3) in its assessment of 3-pentanone.

Response. AWOs incorrectly stated the test rule requirements for 3-pentanone. The proposed 

test rule proposed a bacterial reverse mutation test and a reproduction/developmental toxicity 

screening test. For these proposed studies, EPA assessed existing data for 2-pentanone 

submitted to the HPV Challenge Program. Based on similar chemical/physical properties and 

comparative acute lethality data for 2-pentanone and 3-pentanone, EPA considers a read-across 

approach from 2-pentanone to 3-pentanone to be appropriate, and EPA has determined that the 

toxicity data submitted for 2-pentanone to the HPV Challenge program are adequate to satisfy 

the requirements for bacterial reverse mutation (in vitro) and reproduction/developmental 

toxicity screening testing of 3-pentanone. Therefore, additional testing of 3-pentanone is not 

necessary, and EPA has dropped this chemical from the test rule. 

3. Benzene, 1-chloro-4-(trifluoromethyl)- (PCBTF) (CASRN 98-56-6).

Comment. GCC in their comments provided results of a log Kow study, an aerobic 

biodegradation study and an anaerobic biodegradation study conducted by Lilly Research 

Laboratories and submitted to EPA under a 1983 negotiated testing agreement (NTA, 48 FR 

32730), in which Occidental Chemical Corporation (Occidental) had agreed to a set of testing 

to evaluate the human health effects, environmental effects, chemical fate and 

physical/chemical properties of PCBTF. 

Response. EPA agrees that additional log Kow testing for PCBTF is unnecessary. Although the 

submitted study does not appear to meet GLP requirements, the test itself appears valid and the 

measured log Kow value is close to the one estimated by EPI Suite 4.1 (3.7 vs. 3.6), which lends 

confidence to the results.
16

However; the aerobic and anaerobic biodegradation studies were inconclusive, in part or 

large measure because the lab did not adequately address the substance's volatility, which is 

substantial. Although substantial, it is not so high as to render irrelevant any concern (thus 

testing) relating to media other than air. This means that biodegradation testing should proceed. 

Since the rule specifies ready biodegradability, this endpoint should be determined using a 

guideline appropriate for volatile chemicals. In any case, it would be a good idea for the testing 

lab to run high-performance liquid chromatography (HPLC) analyses and confirm that the test 

substance has been delivered to the test vessels in the amount specified in the guideline as a 

check to be sure that most of it is not lost to the atmosphere somewhere between removing it 

from the supply bottle and getting it into the inoculated test medium. This is what happened the 

first time PCBTF was tested under the NTA. This requires considerable care and even with the 

most commonly used methods for volatiles, one has to inject the substance directly into the 

bottle through a leak-proof septum.

Comment. Regarding the need to conduct acute toxicity studies in fish, invertebrates, and 

plants, GCC provided results of five ecotoxicity studies performed by Occidental and submitted 

to EPA under the 1983 negotiated testing agreement. These ecotoxicity studies address acute 

toxicity in rainbow trout (Hooker, 1979a) and bluegill sunfish (Hooker, 1979b), chronic 

toxicity in fathead minnows (Hooker, 1981a), acute toxicity in Daphnia (Hooker, 1979c), and 

chronic toxicity in Daphnia (Hooker, 1979d). GCC believes that additional toxicity tests in fish 

and Daphnia are not needed because (1) EPA stated in its 1985 Federal Register (50 FR 42216)

notice that “the acute and subchronic environmental effects of [PCBTF] on aquatic species are 

well characterized [and] additional testing is not considered necessary”, and (2) the five studies 

are acceptable based on an evaluation under Section 5 of EPA’s 1999 Guidance for Assessing 

the Adequacy of Existing Data. SMC, GCC, and IsleChem believe that the only data gap for 

PCBTF is algal toxicity. SMC and IsleChem noted that even though a data gap exists, it is 

questionable whether a study can be justified given the extremely low water solubility of 

PCBTF. GCC proposed that the final test rule be modified to require only algal toxicity testing 

(ASTM E 1218-04e1).
17

Response. EPA has reviewed the studies provided by GCC and concluded that the requirements 

to conduct acute studies in fish and invertebrates are satisfied. However, EPA does not agree 

with the assertion of SMC and IsleChem that it is questionable whether a toxicity to aquatic 

plants study can be justified given the extremely low water solubility of PCBTF. If the water 

solubility of PCBTF was great enough to permit studies to be conducted with fish and 

invertebrates, it should be feasible to conduct a toxicity study in algae. Therefore, EPA 

maintains the requirement to conduct a toxicity study in algae.

Comment. Regarding the need to conduct a reproduction/developmental toxicity test, GCC, 

IsleChem and SMC provided results of a two-generation reproductive toxicity study in rats 

performed by Occidental prior to the NTA and submitted to EPA under the NTA (Hooker, 

1981b). GCC, IsleChem and SMC consider this study to be valid and acceptable for the HPV 

Chemical Challenge and OECD SIDS programs. In addition, GCC, IsleChem and SMC noted 

that the National Toxicology Program (NTP) currently has ongoing repeated-dose inhalation 

toxicity and developmental toxicity studies of PCBTF, such that additional testing requirements 

would be duplicative of the NTP studies. 

Response. EPA has reviewed the two-generation study provided by GCC, IsleChem and SMC 

and found it adequate to satisfy the test rule requirement. EPA also agrees with the GCC, 

IsleChem and SMC that the two NTP studies to be conducted, i.e., a teratology inhalation study 

and a continuous breeding inhalation study in rats, will provide sufficient information regarding 

reproductive/developmental endpoints to satisfy the test rule requirement. Therefore, EPA is 

not finalizing the reproduction/developmental toxicity screening test rule proposed for PCBTF.

4. Benzenesulfonic acid, 3-nitro-, sodium salt (1:1) (CASRN 127-68-4).

Comment. AWOs stated that the proposed test rule requires a reproductive/developmental 

toxicity screening test for benzenesulfonic acid, 3-nitro-, sodium salt. AWOs noted that a 

search of TOXLINE identified an abstract from a 1996 report on a safety assessment for this 

chemical (Anonymous, 1996) in which it was stated that a median lethal dose in rats was not 

achieved at an oral dose of 5,000 mg/kg, that chronic administration of a composite hair dye 
18

formulation containing 2.25% of the test chemical did not cause gross or microscopic changes 

in dogs, and that the same hair dye formulation was not classified as a reproductive toxicant in 

rats or rabbits administered the formulation in the diet at concentrations of 0.005% or 0.02%. 

Response. EPA reviewed the study report (Wernick et al., 1975) of chronic, reproductive, and 

developmental toxicity that was summarized in the safety assessment for benzenesulfonic acid,

3-nitro-, sodium salt (sodium m-nitrobenzenesulfonate) mentioned by AWOs. The study 

employed a composite hair dye formulation that included 15 separate dyes and 10 separate base 

substances; sodium m-nitrobenzenesulfonate constituted only 2.25% of the composite 

formulation, thus precluding the usefulness of the study for hazard characterization. The 

requirement for a reproduction/developmental toxicity screening test will be maintained.

Comment. AWOs urged EPA to consider a read-across approach from the sponsored HPV 

program chemicals benzenesulfonic acid (CASRN 98-11-3) and hydroxybenzenesulfonic acid 

(CASRN 1333-39-7) for reproductive/developmental toxicity endpoints.

Response. Regarding AWOs’ suggestion to use a read-across approach from sponsored HPV 

program chemicals, EPA reviewed its comments on mammalian toxicity testing plans for 

benzenesulfonic acid and hydroxybenzenesulfonic acid. EPA determined that mammalian tests 

would not likely provide meaningful systemic toxicity information for these two chemicals, 

which are strong acids. However, the chemical behavior of benzenesulfonic acid, 3-nitro-, 

sodium salt in mammalian systems has not been provided by the AWOs and inadequate data 

are available from which to predict the behavior of benzenesulfonic acid, 3-nitro-, sodium salt

based on available results for benzenesulfonic acid and hydroxybenzenesulfonic acid. 

Therefore, EPA considers data regarding reproductive and developmental toxicity endpoints for 

benzenesulfonic acid, 3-nitro-, sodium salt to be necessary and reaffirms the requirement for a 

reproductive/developmental toxicity screening test for benzenesulfonic acid, 3-nitro-, sodium 

salt.

5. 1-Tetracosanol (CASRN 506-51-4); 1-Hexacosanol (CASRN 506-52-5).
19

Comment. 1-Tetracosanol was included in the proposed test rule based on volumes reported 

under a 2006 Preliminary Assessment Information Reporting Rule (PAIR). The volume 

reported by the only manufacturer, SASOL, under PAIR was 1,980,944 Kg. A comment by 

SASOL indicated that an error was made in converting pounds to kilograms resulting in an 

overstatement of the volume. SASOL recalculated the volume for this chemical as 198,094.4 

Kg. Therefore, the chemical is not a high production volume chemical and is not subject to the 

rule.

Response: EPA has examined the corrected information submitted by SASOL and agrees 1-

tetracosanol is not a high production volume chemical and has not included 1-tetracosanol in 

the test rule. However, EPA may reevaluate this decision based on IUR data to be reported later 

in 2011. In the event IUR or other data provides new or additional support for the TSCA 

section 4(a)(1)(B)(i) finding for 1-tetracosanol, EPA will further evaluate the information 

submitted by SASOL regarding the chemical/physical properties of this chemical substance, 

and, if necessary, EPA will take appropriate steps to proceed with a test rule for this chemical 

substance.

Comment. 1-Hexacosanol was included in the proposed test rule based on volumes reported 

under a 2006 Preliminary Assessment Information Reporting Rule (PAIR). The volume 

reported by the only manufacturer, (SASOL) under PAIR was 974,283 Kg. A comment by 

SASOL indicated that an error was made in converting pounds to kilograms resulting in an 

overstatement of the volume. SASOL recalculated the volume for this chemical as 97,428.3 

Kg. Therefore, the chemical is not a high production volume chemical and is not subject to the 

rule.

Response: EPA has examined the corrected information submitted by SASOL and agrees 1-

hexacosanol is not a high production volume chemical and has not included 1-hexacosanol in 

the test rule. However, EPA may reevaluate this decision based on IUR data to be reported later 

in 2011. In the event IUR or other data provides new or additional support for the TSCA

section 4(a)(1)(B)(i) finding for 1-hexacosanol, EPA will further evaluate the information 
20

submitted by SASOL regarding the chemical/physical properties of this chemical substance, 

and, if necessary, take appropriate steps to proceed with a test rule for this chemical substance.

Comment. SASOL requested that EPA remove 1-tetracosanol and 1-hexacosanol from the 

proposed test rule because SASOL believes that sufficient information is already available in 

order to determine or predict the health or environmental effects of these substances. SASOL

does not believe that the proposed studies are necessary to adequately characterize the effects 

of 1-tetracosanol and 1-hexacosanol and, in some cases, are not technically feasible. SASOL

stated that it imports 1-tetracosanol and 1-hexacosanol as components of mixtures of alcohols, 

not as individual substances; therefore, if testing were to be required, it should be as mixtures 

containing these substances rather than as the individual substances. 

Response. As detailed below, based on data provided by SASOL and AWOs, EPA agrees that 

there is adequate information available to characterize the toxicological properties of 1-

tetracosanol and 1-hexacosanol. EPA notes that using data available for other long chain 

alcohols is a valid read-across approach. This approach is supported by the OECD SIDS Initial 

Assessment Profile of the Long Chain Alcohols Category (April 2006), which concluded that a 

comparison of the SIDS endpoints for the linear alcohols C6–C22 demonstrated a clear inverse 

relationship between chain length and toxicological properties. Therefore, read-across of 

toxicity from C6—C22 alcohols to 1-tetracosanol (C24) and 1-hexacosanol (C26) is acceptable 

as conservative prediction of toxicity. 

Comment. AWOs noted that the proposed test rule requires fish and mammalian acute toxicity 

and combined repeated-dose reproduction/developmental toxicity screening tests for 1-

tetracosanol and 1-hexacosanol which have already been evaluated as part of the Aluminum 

Alkoxides Category. AWOs stated that these chemicals should be removed from the chemical 

list because they have already been thoroughly evaluated as part of the Aluminum Alkoxides 

Category sponsored by The Soap and Detergent Association; AWOs provided a web address 

for the evaluation that was submitted to the HPV Challenge program 

(http://www.epa.gov/HPV/pubs/summaries/almalkct/c16706tc.htm).
21

Response. EPA reviewed the data submitted to the HPV Challenge program for the Aluminum 

Alkoxides Category and determined that long chain alcohols were tested as analogues for the 

aluminum alkoxides. The data obtained for the Long Chain Alcohols in the HPV submission 

for Aluminum Alkoxides Category are the same as the data that SASOL discussed for Long 

Chain Alcohols. Responses to specific comments are addressed below.

Comment. SASOL does not possess results for a chronic toxicity study in Daphnia or toxicity 

study in algae. However, SASOL stated that the aquatic toxicity of alcohols was extensively 

reviewed in the LCOH SIAR(OECD, 2006); some relevant conclusions were: (1) studies of 

acute effects in fish (96-h LC50) for single carbon chain lengths greater than C13-14 in fish 

showed no effects up to the limit of water solubility, (2) studies of acute effects in invertebrates 

(48- to 96-h EC50) for single chain lengths greater than C13 showed that no effects were likely 

up to the limit of water solubility, (3) studies of effects in algae (72-h ErC50) for single chain 

lengths greater than C14 showed that no effects were likely up to the limit of water solubility, 

and (4) chronic studies in invertebrates (21-d NOEC) for single chain lengths greater than C15 

showed no effects expected up to the limit of water solubility. SASOL stated that the chronic 

toxicity studies in Daphnia required making extensive modifications to the standard 

experimental protocol to account for the rapid biodegradation of the alcohols and low 

solubility. SASOL noted that even with the extensive modifications and extreme measures 

taken, significant degradation of the alcohols occurred. SASOL stated that based on the 

experiences with the LCOHs tested, it would be unlikely that a chronic Daphnia study with 1-

tetracosanol and 1-hexacosanol would be technically possible.

Response. Information provided by SASOL and information in EPA’s Screening-Level Hazard 

Characterization for Long Chain Alcohols Category (EPA, 2009) indicates that, for single chain 

lengths >C14 and >C15, respectively, no effects are likely on algal growth rate and on chronic 

exposure in invertebrates up to the limit of water solubility. Therefore, EPA will not require

aquatic toxicity testing (i.e., chronic testing in Daphnia and toxicity to algae).

Comment. SASOL stated that a recent review (Veenstra et al., 2009) of the health effects of the 

LCOH category concluded that all members had low toxicity following acute and repeated 
22

exposures and that they are not mutagenic, carcinogenic, or developmental and reproductive 

toxins. The data were found to support the overall conclusion that there is an inverse 

relationship between chain length and the toxicity of long chain alcohols. SASOL indicated that 

it is reasonable to predict that 1-tetracosanol and 1-hexacosanol might have even lower toxic 

potency than the alcohols in the category for which information on health effects is available. 

According to SNA, the available data can be used as conservative predictors of the health 

effects of 1-tetracosanol and 1-hexacosanol.

Response. EPA agrees that information regarding the LCOH category (C6–C22) can be used as 

a conservative predictor for the health effects of 1-tetracosanol and 1-hexacosanol. EPA’s 

Screening Level Hazard Characterization for the Long Chain Alcohols Category (2009), which 

is based upon the OECD SIAR for long chain alcohols, states that the category is justified on 

the basis of similarities in structural, physicochemical, environmental and health parameters. 

That document also concluded that a clear relationship between chain length and toxicological 

properties (as chain length increased, toxicity decreased) justified read across from shorter 

chain alcohols to category members. 

In addition, SASOL noted that part of the health effects database in the LCOH SIAR 

(OECD, 2006) includes studies of Alcohols, C24-34, which is a mixture of long chain alcohols 

derived from beeswax. This substance is composed of a mixture of long-chain alcohols 

including significant amounts of 1-tetracosanol (13.24%) and 1-hexacosanol (15.34%).

Information from the studies conducted with long chain alcohols derived from beeswax was 

evaluated along with the additional data provided by SASOL below. 

Comment. Regarding the requirement to conduct acute oral toxicity studies for 1-tetracosanol 

and 1-hexacosanol, SASOL noted that the LCOH SIAR (OECD, 2006) cites existing oral LD50

values for linear alcohols (1-octanol to 1-docosanol [C22]) that range from >2,000 mg/kg to 

well over 10,000 mg/kg data; most values represent the maximum administered dose. SASOL

believes that acute oral LD50s for 1-tetracosanol and 1-hexacosanol would fall within this range 

and are more likely to be even higher. SASOL indicated that conducting the proposed studies 
23

would not further the understanding of the low acute toxicity of these substances and would 

result in unnecessary use of animals for testing.

Response. EPA agrees that based on information obtained for the LCOH category, it is 

reasonable to predict that 1-tetracosanol and 1-hexacosanol will have low acute toxicity. EPA’s 

Screening Level Hazard Characterization for the Long Chain Alcohols Category (EPA, 2009) 

concluded that the acute toxicity is low for the category members following oral, dermal, or 

inhalation exposure. These data satisfy the test rule endpoint. Therefore, EPA will not require 

the Mammalian Toxicity - Acute endpoint for 1-tetracosanol and 1-hexacosanol.

Comment. Regarding the requirement to conduct a combined repeated-dose 

reproduction/developmental toxicity screening test, SASOL noted that the LCOH SIAR 

(OECD, 2006) references a number of repeated-dose studies of structurally-related long chain 

alcohols (a table is provided). SASOL stated that these studies provide a sufficient basis to 

predict a low order of repeated-dose toxicity for 1-tetracosanol and 1-hexacosanol. SASOL

indicated that conducting the proposed studies would not further the understanding of the low 

acute toxicity of these substances and would result in unnecessary use of animals for testing.

Response. EPA has evaluated the information presented in the LCOH SIAR (OECD, 2006) and 

confirmed that numerous repeated duration studies have been conducted with alcohols of 

varying carbon chain length, including C24–34 (derived from beeswax). Robust summaries of 

these studies are also available as part of the Aluminum Alkoxides Category. EPA has also 

confirmed that the reliability score of these studies is 1 or 2. 

(Note: A score of 1 equals “reliable without restrictions” and is assigned to 

“studies or data...generated according to generally valid and/or internationally 

accepted testing guidelines (preferably performed according to GLP) or in which

the test parameters documented are based on a specific (national) testing 

guideline...or in which all parameters described are closely related/comparable to 

a guideline method;”

A score of 2 equals “reliable with restrictions” and is assigned to “studies or 

data...(mostly not performed according to GLP), in which the test parameters 

documented do not totally comply with the specific testing guideline, but are 

sufficient to accept the data or in which investigations are described which cannot 
24

be subsumed under a testing guideline, but which are nevertheless well 

documented and scientifically acceptable.”)”( OECD, 2005).)

EPA agrees that based on the information available, it is reasonable to conclude 

that the predicted repeated-dose toxicity NOAELs and LOAELs for 1-tetracosanol and 

1-hexacosanol will fall within the range of values for the tested long chain alcohols. 

Therefore, these data satisfy the test rule endpoint and EPA will not require the 

Mammalian Toxicity-Repeated Dose/Reproduction/Developmental endpoint for 1-

tetracosanol and 1-hexacosanol.

Comment. SASOL stated that the LCOH SIAR (OECD, 2006) includes reference to studies that 

found no significant effects on pregnancy rates, uterine parameters, time to pregnancy, and 

gestation length, and no microscopic changes in the reproductive organs of rats exposed to 1-

dodecanol (C12) and 1-octadecanol (C18). SASOL noted that studies of reproductive 

parameters with other long chain alcohols (C22 and C24-34) in rats and/or dogs for periods of 

up to a year showed these substances to have low reproductive toxic potency and that 

conducting the proposed reproductive studies with 1-tetracosanol and 1-hexacosanol seems 

unnecessary. SASOL also noted that results of developmental toxicity studies referenced in the 

LCOH SIAR (OECD, 2006) indicate that long chain alcohols have no potential for 

developmental effects and embryotoxicity or fetotoxicity (tables are provided).

Response. Based on the evaluation of the information provided by SASOL and of the robust 

summaries of the mentioned studies, the information available is adequate to predict that 1-

tetracosanol and 1-hexacosanol do not exhibit potential for developmental toxicity. This is 

consistent with the conclusion of EPA’s Screening Level Hazard Characterization for the Long 

Chain Alcohols Category (EPA, 2009) that overall, the available reproductive and 

developmental toxicity data for the long chain alcohols indicate no developmental toxicity and 

minimal maternal toxicity with repeated exposure. The information available is found adequate 

to characterize the Mammalian Toxicity-Repeated Dose/Reproduction/Developmental endpoint 

for 1-tetracosanol and 1-hexacosanol and EPA will not require the Mammalian ToxicityRepeated Dose/Reproduction/Developmental endpoint for 1-tetracosanol and 1-hexacosanol.
25

Comment. Regarding the requirement to assess the mutagenicity and clastogenicity of 1-

tetracosanol and 1-hexacosanol, SASOL stated that the LCOH SIAR (OECD, 2006) provides 

pertinent data for a whole range of linear alcohols (including C22 and C24–34) and that the 

results were consistently negative (examples are presented in a table). SASOL noted that the 

category of long chain aliphatic alcohols does not contain any structural elements that are of 

concern for potential mutagenic activity and that there is no reason to predict that 1-

tetracosanol and 1-hexacosanol would deviate from this behavior. SASOL stated that 

conducting the proposed studies would not yield any useful information.

Response. EPA has evaluated the information in the LCOH SIAR (OECD, 2006) and the 

pertinent robust summaries of these studies available as part of the evaluation of the Aluminum 

Alkoxides Category and concludes that the data available are sufficient to predict with 

reasonable certainty that 1-tetracosanol and 1-hexacosanol would not be mutagenic or 

clastogenic. Therefore, EPA will not require the Mammalian Toxicity – Genotoxicity endpoint 

for 1-tetracosanol and 1-hexacosanol.

6. 2-Propenoic acid, 2-carboxyethyl ester (B-CEA) (CASRN 24615-84-7).

Comment. The Society of Chemical Manufacturers and Affiliates (SOCMA) commented on 

behalf of two member companies, Bimax, Inc. and Rhodia, Inc. to question EPA’s addition of 

B-CEA to the proposed test rule. SOCMA indicates that there are only two manufacturers of 

this material, Bimax, Inc and Rhodia, Inc. and the combined production is well below 1 million 

pounds of this material and is not likely to increase significantly. Bimax, Inc and Rhodia, Inc 

have subsequently submitted corrections to their respective 2006 IUR reports that have 

substantiated the claim that the combined production volume for this chemical is well below 1 

million pounds. 

Response: EPA has examined the information on production volumes for B-CEA submitted by 

SOCMA (and IUR corrections submitted directly from Bimax, Inc. and Rhodia, Inc.) and agrees

B-CEA is not a high production volume chemical and has not included B-CEA in the test rule.

However, EPA may reevaluate this decision based on IUR data to be reported later in 2011. In 
26

the event IUR or other data provides new or additional support for the TSCA section

4(a)(1)(B)(i) finding for B-CEA, EPA will take appropriate steps to proceed with a test rule for 

this chemical substance.

7. Benzenesulfonic acid, dimethyl- (CASRN 25321-41-9).

Comment. Nease provided a rationale for using data for surrogate chemicals to satisfy the 

proposed test rule for benzenesulfonic acid, dimethyl- because limited data exist regarding the 

physicochemical and toxicological properties of benzenesulfonic acid, dimethyl-. Two groups 

of surrogates were proposed: (1) several salts of the compound, and (2) acid forms of 

structurally-related aromatic sulfonic acids. The use of salts was considered justified on the 

grounds that in aqueous media, pH >1, aromatic sulfonic acids are expected to be highly 

ionized. Thus, data available for benzenesulfonic acid, dimethyl- salts for many endpoints, in 

particular those studies that may take place in aqueous media at or near pH 7 can be used for 

benzenesulfonic acid, dimethyl-. The use of data on the structurally-related aromatic sulfonic 

acids such as benzenesulfonic acid (0 methyl groups) and p-toluenesulfonic acid (1 methyl 

group) was considered justified on the grounds that the addition of methyl groups to the 

aromatic ring will have only a slight electronic effect on the ring and will also “block” 

substituent sites, thus reducing chemical reactivity (benzenesulfonic acid, dimethyl- has 2 

methyl groups attached to the ring). Nease stated that based on the predicted differences in 

chemical properties among these chemicals, the use of data to fill data gaps among the group is 

warranted.

Nease noted that there are several isomers of this test substance, although the CAS entry

does not specify position of the methyl groups. Citing assessments sponsored by Nease and 

conducted by Toxicology Excellence for Risk Assessment (TERA) and the Consortium for 

Environmental Risk Management (CERM), Nease concluded that all testing requirements have 

been previously addressed. Nease believes different isomers, as well as structural analogues, 

are acceptable surrogates. The study analyses utilized data for xylenesulfonic acid (2,4-

dimethylbenzenesulfonic acid) (XSA) and its salts, due to aqueous solubility and the resulting 

expectation of ionization in aquous media. Nease also provided data available for p-
27

toluenesulfonic acid (pTSA) and benzesulfonic acid (BSA), citing their structural similarity to 

benzenesulfonic acid, dimethyl-. Nease provided data on the physical properties of XSA, 

estimated values from EPISuite v4.0, and measured values for the HPV test substance from 

MSDS provided by Arichem company. Properties of the surrogate compounds also provided by 

Nease. 

Response. EPA has evaluated the data provided by Nease on 2,4-xylenesulfonic acid (2,4-XSA) 

isomer (CASRN: 88-61-9) and other analog substances, including XSA salts. With regard to 

physical/chemical (p-chem) properties, EPA finds that the analogs with fewer than two methyl 

groups are unsatisfactory, as are the data for XSA salts. The 2,4-XSA isomer is a solid that is 

recrystallized from water, whereas the test rule chemical (XSA: CASRN 25321-41-9) is an 

unspecified mixture of isomers and a liquid (Arichem MSDS: 95% pure). Acid salt compounds 

have different p-chem properties from the acid forms. For example: pure 2,4-XSA has a 

melting point of 61 - 62 o C and the salt of XSA (CASRN 1300-72-7), which is a mixture of 

salts of isomers, has a melting point of 27 o C. The test rule chemical (XSA: CASRN 25321-

41-9), which is a mixture of isomers, has a melting point of < 25 o C and is expected to be a 

liquid. The boiling point (or decomposition point) is still needed, as is the vapor pressure. 

(Note: the Arichem MSDS listed a flashpoint of 95 °C, so vapor pressure at an elevated 

temperature is sufficient to allow ignition.) Solubility in water was listed at 100% by Arichem, 

so additional solubility data is not needed, but the log Kow partition coefficient should be 

measured and this test requirement remains in the rule. 

Comment. Nease also provided a number of studies on surrogate chemicals and stated that these 

should be adequate for the ready biodegradation testing endpoint in the rule.

Response. Regarding the test requirement for ready biodegradability, some of the studies cited 

by Nease do not fulfill the ready biodegradability endpoint, primarily because they employ the 

use of a microbial inoculum that has been adapted to biodegrade the test substance. A basic 

principle of ready biodegradation testing is that adapted microorganisms are not used. 
28

However, EPA finds that two studies, one using OECD guideline 301B Modified Sturm 

Test on CASRN 1300-72-7 (Xylene sulfonic acid, sodium salt) and another study using OECD 

301C Modified MITI Test on CASRN 827-21-4 (also described as Xylene sulfonic acid, 

sodium salt) fulfill the ready biodegradation endpoint. Results were 87% biodegradation using

the OECD guideline 301B (Modified Sturm Test), and EPA finds these results acceptable. 

Though there was 0% biodegradation in the OECD 301C Modified MITI test, failure in the 

OECD 301C Modified MITI test alone is not usually taken as indication that a substance is not 

degradable in the environment; this test is very stringent and chemicals that fail often 

biodegrade rapidly in the environment and/or are found to be inherently biodegradable.

Therefore EPA agrees no further Ready Biodegradability testing is needed.

Comment: Nease also stated that the studies provided could satisfy the toxicity testing 

endpoints proposed by EPA.

Response. EPA agrees (see specific comments below). The justification provided by Nease for 

using salts of benzenesulfonic acid, dimethyl and acid forms of structurally-related aromatic 

sulfonic acids as read-across chemicals to fill data gaps for toxicological properties of 

benzenesulfonic acid, dimethyl- is appropriate.

Comment. Regarding the need to collect data on acute toxicity to fish and aquatic invertebrates 

and on toxicity to plants (algae), Nease stated that data are available for sodium and calcium 

xylenesulfonate (salts of benzenesulfonic acid, dimethyl-). Nease provided robust summaries of 

these two chemicals. Nease noted that the test data were measured for aqueous solutions of the 

substances at approximately 40–41% active ingredient and were corrected for 100% active 

ingredient (Table 3 provided by Nease). According to Nease, these data are acceptable as readacross.

Response. As indicated above, sodium and calcium xylene sulfonate are regarded as 

appropriate analogues for benzenesulfonic acid, dimethyl- for read-across purposes. The only 

measured value for benzenesulfonic acid, dimethyl- presented in Table 3 of Nease’s comments
29

is a 24-hour LC50 for acute toxicity to fish of >5 mg/L. Since the study is provided a reliability 

score of 4, it cannot be used to fulfill the test rule requirement (a score of 4 equals “not 

assignable” and is given to “studies or data....which do not give sufficient experimental details 

and which are only listed in short abstracts or secondary literature (books, reviews, etc.,).”

(OECD, 2005). Nease also presented read-across values for acute toxicity to fish, invertebrates, 

and algae obtained with the sodium and calcium salts of benzenesulfonic acid, dimethyl-

(corrected to 100% active ingredient) and provided a link to the summaries (Appendix 6 in

Nease’s Table of Contents). Summaries of these studies are also found in Appendix 5a for the 

sodium salt (SIDS Dossier) and Appendix 5e (SIDS Dossier) for the calcium salt. EPA 

reviewed these summaries and found the studies adequately conducted, and the results 

appropriate for read-across to satisfy the test rule requirement. Therefore, EPA will not finalize 

the test rules for aquatic toxicity for benzenesulfonic acid, dimethyl-.

Comment. Nease noted that very limited mammalian data are available for neat

benzenesulfonic acid, dimethyl-, but there are some data on products that contain 80–94% of 

the chemical. Nease stated that since exposure in the environment would be to the deprotonated 

form, salts (K, Na, Ca, NH4) are used as a surrogate for some mammalian toxicity endpoints. 

Selected data are summarized in Table 4 of Nease. Two acute toxicity LD50s are available, an 

oral LD50 and an intraperitoneal LD50. The oral LD50 of >5,000 mg/kg is included in the 

IUCLID dataset, but few details are available. However, Nease noted that this value is 

supported by reliable LD50s for the salts (summaries presented in a SIDS Dossier).

Response. The lack of detail (i.e., number of animals, vehicle, and duration of the observation 

period) in the study with benzenesulfonic acid, dimethyl- summarized in the IUCLID dataset 

precludes its use to satisfy the endpoint of Mammalian Toxicity–Acute. However, acute studies 

with the calcium and sodium salts summarized in SIDS Dossiers are adequate to satisfy the test 

rule requirement. Therefore, EPA will not finalize the test rule for the mammalian acute 

toxicity endpoint for benzenesulfonic acid, dimethyl-.

Comment. Nease stated that there are no data regarding genetic toxicity for benzenesulfonic

acid, dimethyl-, but data are available for the sodium and calcium salts (summaries presented in 
30

a SIDS Dossier) and for p-toluenesulfonic acid and benzenesulfonic acid (summaries presented 

in IUCLID datasets). Assays for gene mutation and chromosomal aberration were mostly 

negative.

Response. EPA reviewed the summaries of in vitro mutagenicity and clastogenicity tests 

conducted with the sodium salt, in vitro mutagenicity and in vivo clastogenicity tests conducted 

with the calcium salt, in vitro mutagenicity and clastogenicity tests with p-toluenesulfonic acid, 

and an in vitro mutagenicity test with benzenesulfonic acid and concludes that the studies were 

conducted appropriately and as such, can be used to fill the data gap for the test rule 

requirement. Therefore, EPA will not finalize the test rule for Mammalian Toxicity –

Genotoxicity for benzenesulfonic acid, dimethyl-.

Comment. Nease noted that there are no data regarding the repeated-dose toxicity endpoint for 

benzenesulfonic acid dimethyl-, but that data on the various salts could be used as surrogates 

(shown in Table 4 and summaries presented in a SIDS Dossier).

Response. Repeated-dose studies conducted with the sodium salt of benzenesulfonic acid 

dimethyl- listed in Table 4 of Nease include 90-day dermal studies in rats and mice, two-year 

dermal studies in rats and mice, two 90-day dietary studies in rats, and a 90-day dietary study in 

mice. EPA has reviewed the summaries of these studies in the SIDS Dossier and concludes that 

they provide adequate information to satisfy the test rule requirement. Therefore, EPA will not 

finalize the test rule for Mammalian Toxicity – Repeated Dose for benzenesulfonic acid, 

dimethyl-.

Comment. Nease stated that there are no data regarding the Reproductive/Developmental 

Toxicity endpoint for benzenesulfonic acid dimethyl-, but there are relevant data for the salts. 

Nease noted that a 90-day oral feeding study with the sodium salt in rats included an 

examination of the reproductive organs and no effects were reported. In addition, a gestational 

exposure study with the calcium salt in rats reported no treatment-related effects (summaries 

presented in a SIDS Dossier).
31

Response. The gestational exposure study with the calcium salt of benzenesulfonic acid 

dimethyl- and the 90-day dietary studies in rats with the sodium salt of benzenesulfonic acid 

dimethyl- that conducted histological examinations on the reproductive organs of males and 

females satisfy the tests rule requirement. Therefore, EPA will not finalize the test rule for 

Mammalian Toxicity –Reproduction/Developmental for benzenesulfonic acid, dimethyl-.

8. Methanesulfonamide, N-[2-[(4-amino-3-methylphenyl)ethylamino]ethyl]-, sulfate (2:3) 

(CD3) (CASRN 25646-71-3).

Comment. Eastman and I3A stated that CD3 is no longer a high production volume chemical. 

Eastman stated that they have “ceased all manufacture and import of CD3 in 2006 and have no 

plans to restart...at this time." Eastman previously reported CD3 manufacture under 2006 IUR.

I3A stated that CD3 is a material used to process images that have been captured by a 

photosensitive silver halide – i.e., traditional photographic film. I3A commented that the 

volume of CD3 that is currently imported has decreased to an annual amount of 435,851 lbs 

based on 2009 production/import data. None of their member companies have produced CD3 

since 2006. I3A indicates import volumes are well below the threshold, and are continually 

declining. The use of photographic film has been nearly completely replaced by digital 

photography. They add that there is no chance that this trend will be reversed, or that the 

related use of CD3 will increase.

Response: EPA has reviewed the information provided by Eastman and I3A updating the past 

production and importation profile to the present time for CD3. EPA agrees that this substance 

is no longer a high production volume chemical and has not included CD3 in the test rule.

However, EPA may reevaluate this decision based on IUR data to be reported later in 2011. In 

the event IUR or other data provides new or additional support for the TSCA section

4(a)(1)(B)(i) finding for CD3, EPA will take appropriate steps to proceed with a test rule for 

this chemical substance.

Comment. I3A stated that the proposed test rule for CD3 requires screening level data for 

physical/chemical properties, environmental fate and pathways, aquatic toxicity and 
32

mammalian toxicity. I3A believes that existing data on CD3 and a close structural analogue, 

N,N-bis-(2-hydroxyethyl)-p-phenylenediamine (CASRN 54381-16-7), provide the required 

information and help to avoid duplicative testing. I3A provided results of studies for CD3 that 

include physical/chemical properties endpoints (melting point, boiling point, vapor pressure, 

water solubility, and Log Pow), biodegradation, aquatic toxicity endpoints (acute toxicity to fish 

and aquatic invertebrates, toxicity to aquatic plants), and mammalian toxicity endpoints 

(repeated-dose studies). The data were extracted from published studies and internal company 

reports. Data provided for the proposed analogue chemical include repeated-dose toxicity 

studies and developmental and reproductive toxicity studies.

Response. Regarding the endpoints for melting point, log Kow, and water solubility, EPA finds 

the data provided acceptable and is not requiring those testing endpoints in this final rule. 

Regarding the boiling point and vapor pressure endpoints, as a general rule, EPA does not 

accept estimated data for chemical/physical properties in lieu of measured data. However, EPA 

has determined for this chemical that measured data are not required for these endpoints. The 

melting point of 168 - 171 oC with decomposition at 200 oC obviates the need for a boiling 

point determination. In addition, the material is a high melting stable solid and has negligible 

vapor pressure at room temperature, so this test endpoint is also not needed.

However, in regards to toxicity testing, although both the analog and subject test 

substance are based on a p-phenylenediamine substructure, the proposed analogue, N,N-bis-(2-

hydroxyethyl)-p-phenylenediamine (CASRN 54381-16-7), contains structural features that are 

not shared by the test rule substance. The commenter has not addressed whether the differences 

in structure between the two substances (and potentially, in their expected metabolites) would 

influence their toxicological properties. The structural differences in question reside in the side 

chains attached to the respective amines. Specifically, the test rule substance contains one alkyl 

group and one sulfonamide-substituted alkyl group on the amine. In contrast, the proposed 

analogue contains two hydroxyethyl groups in these positions. The differences in the electronic 

character of these two different types of functional groups (the sulfonamide is electronwithdrawing, while the alcohols are electron-donating) can influence how these chemicals will 

interact in biological systems, including enzyme active sites. In addition, the hydroxyethyl 
33

groups have the potential to undergo metabolic oxidation to their corresponding carboxylic 

acids. A potential metabolic product from this pathway is a bis(carboxymethyl)amine, which 

would have the potential to chelate ions of biological significance, such as calcium, 

magnesium, iron and zinc. In the absence of any discussion concerning how and why the data 

generated for the proposed analogue could be expected to accurately represent the toxicity of 

the test rule substance, the use of the proposed analogue is not supported. 

With respect to the required toxicity testing, the commenter has provided a range of 

aquatic toxicity studies conducted using CD3 as the test substance, including two 96-hour acute 

toxicity to fish studies, both performed in fathead minnow (Primephales promelas), a 96-hour 

acute toxicity to Daphnia study, and a toxicity to aquatic plants (algae) study. However, the 

method section of the summary of one study indicates that the fish were tested in containers 

which also held snails, water fleas, flatworms, segmented worms, side swimmers, and pill bugs 

as part of a “multispecies screening test.” This method of testing deviates from acceptable 

testing methods for the acute toxicity to fish endpoint and is, therefore, not acceptable in 

fulfillment of the test rule for acute toxicity to fish testing. It is not clear what impact the 

presence of other organisms might have on the toxicity of the test substance to the fish. The 

other study, conducted as an acute, flow-through toxicity study, appears to be acceptable, with 

a 96-hour LC50 of 3.6 mg/L obtained for the test substance. Therefore, the need for acute 

toxicity to fish testing has been met by this study.

However, the acute toxicity to Daphnia study, has the same weakness as the first acute 

toxicity to fish study described above; i.e., the Daphnia were tested in the direct presence of 

other aquatic organisms. Also, the acute toxicity to aquatic plants (algae) study used an 

exposure period of 7 days; the test protocol for the test rule requires a 96-hour exposure. 

Therefore, if this substance had been included in the final rule, the chronic toxicity to Daphnia

and toxicity to aquatic plants testing requirements for CD3 would still be required.

Two repeated-dose toxicity studies were submitted for which the testing was performed 

using CD3. Both of these tests were performed in hamsters; one was a study conducted for 27.5 

months with the test substance delivered by gavage; the other was a 2-year cancer bioassay 
34

with the test material delivered in feed. Although these tests could be considered adequate to 

cover the requirement for repeated-dose testing, the only reproductive/developmental testing 

results were supplied on the proposed analogue compound. Since, as explained above, testing 

on the analog is not considered acceptable for filling the requirements of the test rule for CD3, 

if this test substance had been included in the final rule, reproductive/developmental testing 

would still be required. Moreover, since the repeated-dose testing was performed on hamsters, 

it is considered preferable that the requirement for mammalian toxicity testing be performed on 

rats using the combined repeated dose toxicity study with the reproduction/developmental 

toxicity screening test: 40 CFR 799.9365. 

In summary, if CD3 met the finding of “substantial production,” aquatic toxicity testing 

would still be required for CD3 as indicated by the proposed test rule, although further acute 

toxicity to fish testing would not be required. Acute toxicity to Daphnia testing and acute 

toxicity to aquatic plants (algae) testing would also still be required. In addition, mammalian 

toxicity-repeated dose/reproduction/developmental testing would still be required. If conducted, 

all of these tests should be conducted using CD3 as the test substance.

9. 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) (CASRN 

52556-42-0).

Comment. GE submitted comments requesting exemption from the final rule based on 

technical information provided to EPA that their production does not create any substantial 

risk to human health or the environment based on exposure. The case for inclusion in this test 

rule is based on total production volume for all manufacturing sites. GE is not claiming they 

no longer manufacture the chemical. They are only stating that they think there is limited 

exposure from their manufacturing process. GE submitted comment requesting exemption from 

test rule because use/processing does not result in signifcant risk to humans or environment.

Response: GE contributes to a national aggregate of more than 1 million pounds. The 

exposure criteria (general population, consumers, workers) are also met based on aggregate 
35

information. Therefore, this chemical will remain in the final test rule, and GE, as a 

manufacturer of this substance is also subject to the final test rule.

10. Coal/Coal Tar Chemical Substances: 

 Solvent naphtha (coal) (CASRN 65996-79-4);

Tar oils, coal (CASRN 65996-82-9);

Tar, coal, high-temperature (CASRN 65996-89-6);

Distillates (coal tar) (CASRN 65996-92-1);

Pitch, coal tar-petroleum (CASRN 68187-57-5);

Extract residues (coal), tar oil alk., naphthalene distn. residues (CASRN 73665-18-6).

Comment. ACCCI considers the six coal tar-derived chemicals included in the proposed test 

rule to represent a category of similar substances because they are all derived from coal tar and 

are considered to share similar chemical compositions and properties. Therefore, ACCCI 

considers coal tar products such as coal tar creosote, coal tar, and a large number of other coal 

tar products (some of which are undergoing evaluation under the European Registration, 

Evaluation, Authorization and Restriction of Chemicals [REACH] program) to be analogues to 

the six coal tar-derived test rule substances. ACCCI proposed a read-across approach between 

the coal tar-derived test rule substances and the analogues to predict chemical and toxicological 

properties for those substances without readily available data. ACCCI submitted compositional 

data for the six coal tar-derived test rule substances and for coal tar creosote (one of the 

proposed analogues). ACCCI included a table (Table 2.4) that presents information regarding 

proposed analogues for the coal tar-derived test rule substances based on similarities in the 

distillation process, physical properties, and chemical composition. ACCCI stated that the 

submission of coal tar products to the REACH program likely includes relevant information, 

but that time constraints of the proposed test rule deadline made it impossible to obtain this 

information and assess it for relevance. ACCCI stated that no further testing of coal tar-derived 

test rule substances should be initiated prior to evaluation of data for coal tar substances 

submitted to the REACH program and evaluation of data for coal tar petroleum pitch submitted 

to the HPV program as Heavy Fuel Oils Category.
36

Besides the above comments submitted on May 26, 2010, ACCCI submitted an 

additional set of comments on July 29, 2010. In these comments ACCI requests that the above 

six chemicals be removed from the test rule. This request is based on the results of an 

independent survey commissioned by ACCI to provide accurate and reliable estimates of 

current exposure to the six coal-derived chemicals. The survey concludes that there is not 

adequate support for a finding of significant or substantial exposure for any of the six 

substances. The commissioned report concludes that the discrepancy between information on 

consumer and commercial use contained in EPA’s Exposure Findings Document and the 

commissioned survey results appear to be due in part, to mistakes in reporting that may have 

been reflected in the 2006 IUR reports filed by certain facilities.

Response: EPA read and evaluated the comments and all substantive information submitted by 

ACCCI including the independent commissioned survey of all manufacturers and processors of 

the six coal-derived chemicals. EPA also asked for additional supporting information from the 

individual companies. The companies provided this information. Based on the confirmatory 

information provided by the companies, EPA agrees that these substances no longer meet the 

production (tar, coal, high temperature) or exposure-based findings (all six coal-tar derived 

chemical substances) for requiring testing under TSCA section 4(a)(1)(B). Therefore, EPA will 

not include these chemicals in the test rule. However, EPA may reevaluate this decision based 

on IUR data to be reported later in 2011. In the event IUR or other data provides new or 

additional support for the TSCA section 4(a)(1)(B)(i) finding for any of these six coal-tar 

derived chemical substances, EPA will take appropriate steps to proceed with a test rule for the 

chemical substance(s).

EPA notes that the analogues proposed for this submission appear to have been chosen 

on the basis of process similarities with the six test rule substances. While it is reasonable to 

expect that similar process will produce similar substances, the only process information 

available consists of the CAS definitions associated with the CAS registry numbers for the test 

rule substances and the proposed analogues. These definitions are broadly written and often do 

not provide enough information to assess the similarities of these streams. The suitability of 

any proposed analogue must be verified by comparing its compositional similarity with its 
37

corresponding target stream. Although it is understood that coal tar distillates and derivatives 

are highly variable, complex mixtures, nearly half of the constituents of two of the six test rule 

substances (coal tar, high temperature and coal tar distillates) are unidentified, and nearly 88% 

of the constituents of a third substance (coal tar-petroleum pitch) are unidentified. Minimal 

compositional information is provided for the proposed analogues, so comparison to the test 

rule substances is impossible. 

In the absence of compositional data to allow direct comparison of these streams, 

similarities in measured data, such as physical properties, may help support a claim that two 

substances can possess similar characteristics. However, there are differences in physical 

properties (notably boiling range and vapor pressure) between some of the test rule substances 

and their corresponding proposed analogues that appear to signal significant differences in 

chemical composition. Therefore, the use of the analogues proposed by ACCCI is inadequately 

supported in the submission due to poor characterization of the streams involved and 

differences in their reported physical properties. 

EPA does, however, agree with the commenters with respect to not needing additional 

testing for ready biodegradability. Although ready biodegradability data for the 6 subject

substances (including 73665-18-6) was not provided, the commenter did, however, provide 

ready biodegradability data for 5 individual components and inherent biodegradability data for 

one individual component of the subject substances from the Japanese MITI biodegradation 

and accumulation database. EPA typically judges studies from this database to be acceptable. 

These data indicate the majority of the components are not readily biodegradable. It follows 

that mixtures of these substances or mixtures containing these substances as a major component 

are not likely to be readily biodegradable. The following statement is made by ACCI on page 

16 of their comment document:

“Based on limited standard ready biodegradation data (Table 2.6) and the relatively limited 

solubility of the coal tar substances (Table 2.5), it is expected that biodegradation of these 

substances would be minimal, and it is unlikely that further testing would alter this conclusion.”
38

This statement acknowledges that the substances can be classified as not readily biodegradable. 

EPA agrees with this statement, does not think additional ready biodegradability testing is 

necessary, and is therefore dropping that test endpoint for these six substances. 

Specific comments regarding the six test rule streams and their corresponding proposed 

analogues are detailed below.

Solvent naphtha (coal), CASRN 65996-79-4: 

Proposed analogue: Distillates (coal tar), light oils, CASRN 84650-03-3

Solvent naphtha is one of the better characterized of the streams (92.3% of the constituents are 

identified in Table 2.1), with indene identified as the major component (58.5%). The 

composition of the proposed analogue stream is not provided. The boiling point ranges reported 

in the CAS definitions for these two streams and reported in Table 2.4 appear to be comparable. 

However, the boiling ranges reported in Table 2.5 are significantly different: 83-87 °C for 

solvent naphtha and 230-500 °C for coal tar distillates light oils. This difference suggests that 

the vapor pressure of solvent naphtha could be several orders of magnitude higher than that 

reported for the proposed analogue (0.005 torr).

Tar oils, coal, CASRN 65996-82-9:

Proposed analogue: Distillates (coal tar), light oils, CASRN 84650-03-3

83.9% of the constituents of coal tar oils are identified in Table 2.1, with the major component 

identified as naphthalene (45.94%). The composition of the proposed analogue stream is not 

provided. The boiling point ranges reported in the CAS definitions for these two streams and 

reported in Table 2.4 appear to be comparable. However, the boiling ranges, vapor pressure 

ranges and water solubilities reported for these two streams in Table 2.5 differ. Differences in 

physical properties are likely to reflect compositional differences between these two streams, 

which weaken support for using data for the proposed analogue to characterize coal tar oils.

Tar, coal, high-temp., CASRN 65996-89-6:

Proposed analogue: Tar, coal, CASRN 8007-45-2
39

Approximately 50% of the constituents in high-temp coal tar are identified, with the major 

component identified as naphthalene (12.29%). The composition of the proposed analogue 

stream is not provided. The vapor pressures reported in Table 2.5 suggest that the proposed 

analogue may contain more volatile components than the parent stream.

Distillates (coal tar), CASRN 65996-92-1:

Proposed analogue: Anthracene oil, CASRN 90640-80-5

Proposed analogue: Distillates (coal tar) heavy oils, CASRN 90640-86-1

Approximately 55% of the constituents of coal tar distillates are identified, of which 

benzo(f)quinoline is the principle constituent (12.52%). No compositional information is 

provided for either of the two proposed analogue streams. The CAS registry file definition for 

anthracene oil identifies its primary constituents as phenanthrene, anthracene and carbazole. 

According to the compositional information in Table 2.1, phenanthrene comprises 3.3% of coal 

tar distillates, carbazole comprises 1.83% and anthracene is absent. The available information 

does not sufficiently demonstrate compositional similarity among these three process streams.

Pitch, coal tar-petroleum, CASRN 68187-57-5:

Proposed analogue: Pitch, petroleum, arom., CASRN 68187-58-6

Proposed analogue: Pitch, coal tar, high-temp., CASRN 65996-93-2

This is the least characterized of the test rule streams submitted. Only 12.1% of the constituents 

have been identified, with benzo(a)pyrene as the primary component at 1.27%. Compositional 

information is not provided for the two proposed analogue streams. The vapor pressures 

reported in table 2.5 for the two analogues differ from each other by at least 4 orders of 

magnitude, suggesting that these streams may not be that similar to each other, and their 

similarity to the test rule stream is not apparent. In addition, the source(s) for this stream are not 

clearly described, making comparison to the proposed analogues difficult if not impossible.

Extract residues (coal), tar oil alk., naphthalene distn. residues, CASRN 73665-18-6:

Proposed analogue: Creosote, CASRN 8001-58-9

Approximately 80% of the constituents of this stream have been identified, with acenaphthene 

as the primary constituent (13.91%). Partial justification for the use of this analogue is 
40

presented in Table 2.2; however, the compositional information is presented on the basis of 

broad chemical categories rather than individual constituents.

Furthermore, ACCCI’s read-across strategy for toxicological endpoints is not clearly 

explained. ACCCI presents toxicity data for various coal tar substances and coal tar 

components, but does not synthesize these data into a comprehensive table which demonstrates 

how available data will be read across to the six test-rule substances for each required endpoint. 

Data for each of the required endpoints are not available for each of the proposed analogues in 

ACCCI’s Table 2.4, yet ACCCI claims that almost all of the test-rule required endpoints can be 

satisfied via read-across of data from “comparable substances” (ACCCI’s Table 2.19).

Comment. ACCCI considers measured data for primary constituents of coal tar substances (e.g., 

indene or naphthalene) to be appropriate for read across to coal tar-derived test rule substances 

where data are not readily available.

Response. In addition to the proposed analogue process streams already discussed, the 

submitter presented physical property and/or fate data for 11 single chemicals that have been 

identified in the test rule streams. In general, single chemicals do not adequately represent 

complex process streams because they do not account for interactions between different 

constituents within mixtures. As shown in the table below, the different test rule streams 

contain varying amounts of these individual chemicals. Together, the single chemicals 

comprise between 3.2% and 72.7% of the six test rule stream components. This information 

suggests that data on the primary constituents of coal tar substances are not adequate for readacross to coal tar-derived test rules substances.

Component Coal-derived process streams, CASRNs

65996-79-4 65996-82-9 65996-89-6 65996-92-1 68187-57-5 73665-18-9

1-Me 

naphthalene

0 2.20 0.80 0 0 5.30

2-Me 

naphthalene

0 5.90 1.60 0.13 0 13.70

Indene 58.50 3.00 1.30 0 0 0
41

Acenapthene 0 5.39 0.48 0 0.04 13.91

Anthracene 0 1.81 1.63 0 0.11 3.82

Benzo(a)pyrene 0 0 0.60 0.98 0.77 0.10

Fluoranthene 0 1.21 3.23 0 1.02 2.48

Fluorene 0 0 1.70 0 0 8.70

Naphthalene 1.37 45.94 12.29 0.01 0 1.18

Phenanthrene 0 6.51 4.50 3.30 0.37 11.74

Pyrene 0 0.76 2.42 9.41 0.92 1.69

Totals 59.87 72.72 30.55 13.83 3.23 62.62

Comment. ACCCI emphasized the fact that each of the coal tar substances is the product of a 

process that includes variability in feedstock and distillation methods; thus each coal tar 

substance is inherently a category of similar but not identical mixtures. ACCCI noted that 

testing of a given substance would obviate the need to infer that small differences in 

composition do not markedly alter toxicological properties.

Response. As indicated above, ACCCI did not provide adequate information documenting that 

the differences in composition between coal tar substances are small. Therefore, it is not clear 

that differences in toxicological properties between coal tar substances would be small.

Comment. ACCCI stated that no two coal tar substances are identical in their chemical 

composition, but that the constituent compounds that contribute to aquatic toxicity are 

generally the same, which indicates that toxicity data for the major constituents can generally 

be used in evaluating aquatic toxicity of the coal tar substances. ACCCI presented aquatic 

toxicity data for major coal tar creosote constituent compounds intended to be representative of 

the aquatic toxicity hazard of the six coal tar-derived test rule substances.

Response. No evidence was provided to suggest that toxicity data from major coal tar creosote 

constituent compounds would adequately represent the toxicity of the six coal tar-derived test 

rule substances. As indicated above, information on the chemical composition of coal tar 

substances that was provided does not adequately indicate that the coal tar substances are 

sufficiently similar to use as analogues for one another. The log Kow and water solubility values 
42

for “primary components” of coal tar substances, as shown in Table 2.5 of ACCCI’s comments, 

vary greatly. The log Kow of individual components ranges from 2.92 to 5.97, while the log Kow

of the proposed analog creosote is 1.0; because log Kow influences aquatic toxicity, the toxicity

of individual components may not be representative of the toxicity of the coal tar mixtures. 

Water solubility of individual components ranges from insoluble to 31 mg/L, while the water 

solubility of the six coal tar test-rule substances ranges from insoluble to slightly soluble; again, 

these differences suggest that the toxicity of individual substances will not be representative of 

the toxicity of the coal tar mixtures.

Comment. The proposed test rule requires aquatic toxicity testing for each of the six coal tarderived test rule substances. For the acute toxicity to aquatic invertebrates endpoint, ACCCI 

provided information for solvent naphtha (coal) (CASRN 65996-79-4) regarding the acute 

toxicity to aquatic invertebrates (48-hour EC50 of 1-6 mg/L for daphnids) and vertebrates (96-

hour LC50 of 20-50 mg/L for fathead minnows) and cited the information to a European 

Commission document titled “Solvent naphtha (coal) (CASRN 65996-79-4): IUCLID data set” 

dated February 19, 2000 (European Commission, 2000). ACCCI also provided acute aquatic 

invertebrate and vertebrate toxicity data for selected coal tar substances considered to be 

analogues to the test rule substances, and for selected coal tar polycyclic aromatic hydrocarbons 

(PAHs). ACCCI considered the acute toxicity to aquatic invertebrates and vertebrates endpoints 

to be satisfied using data from solvent naphtha, analogue data, PAH data, and a read-across 

approach.

Response. EPA retrieved the IUCLID dataset for solvent naphtha (European Commission, 

2000), as well as available IUCLID datasets for substances considered by ACCCI to be 

analogues to the coal tar-derived test rule substances. For solvent naphtha, the ACCCI and 

IUCLID data set summaries provided insufficient information from which to assess the 

adequacy of the studies that evaluated acute toxicity to aquatic invertebrates and vertebrates 

endpoints. The IUCLID data set (European Commission, 2000) cited Aral Aktiengesellschaft 

Bochum as the source of study information, but provided no additional reference information. 

Additional study details would facilitate a determination of study adequacy. EPA has 

determined that the a category approach for the six coal tar-derived test rule substances 
43

suggested by ACCCI is not acceptable based on compositional and physical/chemical 

differences between these substances. EPA has also determined that compositional and 

physical/chemical differences among ACCCI-suggested analogue substances and coal tarderived test rule substances precludes the usefulness of a read-across approach either between 

the test rule substances or from the proposed analogue substances and the test rule substances. 

If these substances had been included in the final rule, the acute toxicity to aquatic invertebrates 

endpoint for each of the six coal tar-derived test rule substances would still be required.

Comment. For the chronic toxicity to aquatic organisms endpoint, ACCCI cited a 21-day fish 

study that reported a LOEC of 1.9 mg/L for coal tar, one of the substances that ACCCI 

considers to represent an analogue to the coal tar-derived test rule substances. ACCCI also 

provided results from tests of selected individual PAHs in fish and aquatic invertebrates. For 

the toxicity to aquatic plants endpoint, ACCCI provided summary data for algae exposed to 

high temp coal tar pitch and for algae, macrophytes and duckweed exposed to coal tar creosote, 

substances considered by ACCCI to be analogues to the coal tar-derived test rule substances. 

ACCCI considered the chronic toxicity to aquatic organisms and toxicity to aquatic plants 

endpoints satisfied based on a read-across approach.

Response. EPA has determined that compositional and physical/chemical differences among 

ACCCI-suggested analogue substances, individual PAHs, and coal tar-derived test rule 

substances precludes the usefulness of a read-across approach. If these substances had been 

included in the final rule, the chronic toxicity to Daphnia and toxicity to aquatic plants testing 

requirements for each of the six coal tar-derived test rule substances would still be required..

Comment. ACCCI stated that several microcosm studies have been performed on coal tar 

creosote and that results of these studies can be used to supplement the data from the standard 

aquatic toxicity tests for which data were presented.

Response. Data presented from microcosm studies do not satisfy the aquatic toxicity endpoints 

specified in the test rule. Furthermore, EPA has determined that compositional and 
44

physical/chemical differences among ACCCI-suggested analogue substances and coal tarderived test rule substances precludes the usefulness of a read-across approach.

Comment. For the mammalian acute toxicity endpoint, ACCCI presented summary acute oral 

toxicity data for solvent naphtha (CASRN 65996-79-4), for coal tar creosote (CASRN 8001-

58-9) and coal tar pitch (CASRN 65996-93-2) which are considered by ACCCI to be analogue 

substances for the coal tar-derived test rule substances, and for primary components of coal tar 

substances that include indene (CASRN 95-13-6), naphthalene (CASRN 91-20-3), 2-

methylnaphthalene (CASRN 91-57-6), 1-methylnaphthalene (CASRN 90-12-0), phenanthrene 

(CASRN 85-01-8), pyrene (CASRN 129-00-0), fluoranthene (CASRN 206-44-0), and 

benzo[a]pyrene (CASRN 50-32-8). ACCCI also presented summary data for acute inhalation 

toxicity of solvent naphtha, creosote, and indene. ACCCI indicated that the provided data 

satisfy the acute oral toxicity endpoint based on a read-across approach among the six coal tarderived test rule substances and between analogue substances, primary components of coal tar 

substances, and the test rule substances.

Response. EPA reviewed the acute oral and inhalation toxicity information provided by 

ACCCI. Acute toxicity data in the IUCLID dataset for solvent naphtha (European Commission, 

2000) were insufficiently summarized for the purpose of assessing study adequacy. The 

European Commission, 2000) cited Aral Aktiengesellschaft Bochum as the source of study 

information, but provided no additional reference information. Additional study details would 

facilitate a determination of study adequacy. Based on insufficient study details provided for 

solvent naphtha and the determination by EPA that compositional and physical/chemical 

differences among ACCCI-suggested analogue substances and coal tar-derived test rule 

substances precludes the usefulness of a read-across approach. If these substances had been 

included in the final rule, the acute oral toxicity testing requirement for each of the six coal tarderived test rule substances would still be required.

Comment. For the repeated-dose toxicity endpoint, ACCCI presented summary data from oral 

studies of mice administered coal tar (CASRN 8007-45-2; considered by ACCCI to be an 

analogue for the coal tar-derived test rule substances) orally for 15 days (Weyand et al., 1991), 
45

28 days (Culp and Beland, 1994), 185 days (Weyand et al., 1994), 260 days (Weyand et al., 

1995), or 2 years (Culp et al., 1996, 1998). ACCCI also presented summary data from an oral 

study of rats administered a substance described by the study authors as manufactured gas plant 

residue (a type of coal tar formed as a byproduct of coal gasification) for 5 weeks (Chadwick et 

al., 1995). ACCCI identified this substance as coal tar (CASRN 0887-45-2). ACCCI indicated 

that the submitted summaries satisfy the repeated-dose toxicity endpoint for the six coal tarderived test rule substances based on a read-across approach.

Response. EPA reviewed the studies that were summarized by ACCCI for the repeated-dose 

toxicity endpoint. The studies were designed to assess body weight effects, carcinogenicity, and 

DNA adduct formation. The 185-day oral study of Weyand et al. (1994) was the only study to 

report information regarding incidences of nonneoplastic lesions and the histopathological 

examinations for nonneoplastic lesions were performed only on control and high-dose animals 

sacrificed after 95 or 185 days of treatment. EPA determined that the studies for the repeateddose toxicity endpoint that were summarized by ACCCI did not adequately assess 

comprehensive noncancer endpoints at multiple dose levels. Furthermore, EPA determined that 

compositional and physical/chemical differences among ACCCI-suggested analogue 

substances and coal tar-derived test rule substances precludes the usefulness of a read-across 

approach. If these substances had been included in the final rule, the repeated-dose toxicity 

testing requirement for the coal tar-derived test rule substances would still be required.

Comment. The proposed test rule requires in vitro mutagenicity testing for each of the six coal 

tar-derived test rule substances. ACCCI reported the results of a bacterial mutagenicity assay 

for tar, coal, high-temperature (CASRN 65996-89-6; one of the coal tar-derived test rule 

substances). ACCCI also provided results of bacterial mutagenicity assays of substances 

considered to be analogue coal tar substances: coal tar (no CASRN supplied by ACCCI), coal 

tar identified as crude tar, pitch, oils (no CASRN supplied by ACCCI), coal tar pitch volatiles

(no CASRN supplied by ACCCI), and coal tar creosote (CASRN 8001-58-9). In each assay, 

the test substance was positive for mutagenicity in the presence of metabolic activation. ACCCI 

indicated that the provided data satisfy the in vitro mutagenicity endpoint for the coal tarderived test rule substances based on a read-across approach.
46

Response. EPA considers the requirement for in vitro mutagenicity testing of tar, coal, hightemperature (CASRN 65996-89-6) to be satisfied by the information provided by ACCCI for 

this test rule substance. EPA has determined that compositional and physical/chemical 

differences among ACCCI-suggested analogue substances and coal tar-derived test rule 

substances precludes the usefulness of a read-across approach to the remaining five coal-tar test 

rule substances, and therefore these data are not acceptable substitutes for the data that would 

be obtained from a test rule for these five chemical substances. Therefore, if these substances 

had been included in the final rule, this testing would still be required.

Comment. The proposed test rule requires in vitro or in vivo chromosomal damage testing of 

each of the six coal tar-derived test rule substances. ACCCI cited secondary sources (ATSDR, 

2002; WHO, 2004) and three primary sources (Bender et al., 1988; Burgaz et al., 1998; Yadav 

and Seth, 1998) that reported positive results in assays of workers exposed to coal tar 

substances; the results included increased frequencies of chromosomal abnormalities, sister 

chromatid exchanges, and micronuclei. ACCCI also reported increased DNA adduct levels in 

mice administered coal tar or coal tar waste (Culp and Beland, 1994; Culp et al., 1966; Weyand 

et al. 1991, 1994). ACCCI cited US DOE (1994) as a source for reported coal tar-induced 

increased frequency of micronuclei in mammalian V79 cells. ACCCI noted the absence of 

genotoxicity data for solvent naphtha, but indicated that the primary component (indene) was 

negative for mutagenicity in an Ames test using S. typhimurium strains TA98, TA100, TA1535, 

and TA1537 (Curvall et al., 1982). ACCCI stated that based on (1) the current classification of 

coal tar and some PAHs as carcinogens and (2) numerous available in vitro and in vivo assays, 

additional genotoxicity assays are not warranted for the coal tar-derived test rule substances, 

with the possible exception of solvent naphtha.

Response. Chromosomal damage reported in the workers exposed to coal tar substances was 

not directly attributable to the coal tar substances since exposure to other potentially genotoxic 

chemicals occurred in these workers. The increased frequency of micronuclei in V79 cells was 

induced by a substance described by the study authors as low-temperature coal tar, which is not 

one of the six coal tar-derived test rule substances. EPA determined that compositional and 
47

physical/chemical differences among ACCCI-suggested analogue substances and coal tarderived test rule substances precludes the usefulness of a read-across approach. If these 

substances had been included in the final rule, the chromosomal damage testing requirement for 

the six coal tar-derived test rule substances would still be required.

Comment. The proposed test rule requires repeated-dose and reproduction/developmental 

toxicity screening tests for solvent naphtha (coal) (CASRN 65996-79-4); tar oils, coal (CASRN 

65996-82-9); tar, coal, high-temperature (CASRN 65996-89-6); pitch, coal tar-petroleum 

(CASRN 68187-57-5); and extract residues (coal), tar oil alk., naphthalene distn. residues 

(CASRN 73665-18-6). The proposed test rule also requires a reproduction/developmental 

toxicity screening test for distillates (coal tar) (CASRN 65996-92-1). ACCCI presented 

summary data from studies of rats exposed to coal tar liquid during gestation (Hackett et al., 

1984; Springer et al., 1986). ACCCI stated that the composition of the coal tar liquid samples 

used in these studies was “relatively similar to coal tar distillates (CASRN 65996-92-1), extract 

residues (CASRN 73665-18-9), and high temperature coal tar (CASRN 65996-89-6). ACCCI 

also provided summaries of studies in which rats or rabbits were exposed to coal tar creosote

(CASRN 8001-58-9) during gestation (York, 1994, 1995a, 1995b) and a study in which rats 

were exposed to coal tar creosote for two generations (Marcinowski, 1993). ACCCI indicated 

that these data preclude the need for additional repeated-dose and reproduction/developmental 

toxicity testing of the six coal tar-derived test rule substances based on a read-across approach.

Response. EPA determined that compositional and physical/chemical differences among 

ACCCI-suggested analogue substances and coal tar-derived test rule substances precludes the 

usefulness of a read-across approach. If these substances had been included in the final rule,

repeated-dose and reproduction/developmental toxicity testing for the coal tar-derived test rule 

substances would still be required.

Comment. ACCCI stated that, should any further testing for repeated-dose and 

reproductive/developmental toxicity endpoints be required, it should be conducted in a stepwise 

manner. As an example, ACCCI indicated that should a reproduction/developmental toxicity 

screening test for coal tar distillates (one of the six coal tar-derived test rule substances) be 
48

conducted and that results similar to those from existing studies of coal tar and creosote would 

strengthen the proposed read-across approach. ACCCI also indicated that if EPA were to deem 

an additional combined repeated-dose reproduction/developmental toxicity screening test 

necessary, such a study should be performed using solvent naphtha (one of the coal tar-derived 

test rule substances) due to its differences in composition from the other coal tar-derived test 

rule substances.

Response. As noted above, EPA determined that compositional and physical/chemical 

differences among ACCCI-suggested analogue substances and coal tar-derived test rule 

substances precludes the usefulness of a read-across approach. The potential acceptability of a 

“stepwise” scheme for testing these chemical substances would need more evidence to support 

their alleged similarity than was provided by ACCCI in their comments. Given additional 

supporting data, EPA would be willing to consider such an approach.

11. 1,4-Benzenedicarboxylic acid, 1,4-dimethyl ester, manuf. of, by-products from (CASRN 

68988-22-7).

Comment: INVISTA submitted comments claiming minimal potential for release or exposure to 1,4-

Benzenedicarboxylic acid, 1,4-dimethyl ester, manuf. of, by-products from, and on that basis 

no longer meets the exposure-based finding for requiring testing under section 4.

Response. EPA has reviewed the information provided by INVISTA and agrees that this 

substance no longer meets the environmental release or human exposure B-finding thresholds. 

Therefore, EPA is not including this chemical in the final test rule. However, EPA may 

reevaluate this decision based on IUR data to be reported later in 2011. In the event IUR or 

other data provides new or additional support for the TSCA section 4(a)(1)(B)(i) finding for 

this chemical substance, EPA will take appropriate steps to proceed with a test rule for 1,4-

Benzenedicarboxylic acid, 1,4-dimethyl ester, manuf. of, by-products from.

Comment: INVISTA also noted that it has completed 10 of the 12 studies identified in the 

proposed test rule and submitted final and draft reports to EPA on December 3, 2010. 
49

INVISTA also stated that it has not undertaken two of the proposed studies (chronic toxicity to 

Daphnia and combined repeated dose toxicity study) because it believes they are no longer 

necessary for purposes of characterizing the substance. INVISTA stated that it has already 

conducted an acute toxicity test to Daphnia using the OECD 202 method. INVISTA believes 

the results of this study serve as a screening level ecotoxicological evaluation similar to that 

performed for many other HPV chemicals. Thus, INVISTA contends that a chronic study on 

Daphnia is not necessary. INVISTA also commented that the combined repeated dose toxicity 

study is also not necessary because such studies normally would be requested for chemicals 

with potential widespread distribution in the environment. INVISTA indicated that DMT 

residue does not have the potential for widespread distribution in the environment.

Response: The proposed testing included melting point, boiling point, vapor pressure, water 

solubility and log Kow. With respect to these data submitted by INVISTA:

For the melting point determination, the use of differential scanning calorimetry and 

thermogravimetric analysis were performed using OECD guideline 102. After analysis of the 

data, it was found that this residue had a two glass transition states at -20C and 36C. Once the 

temperature rose above 50C, decomposition was observed. EPA finds these data acceptable and 

is not requiring melting point. Since decomposition was observed as the temperature rose above 

50C as noted, EPA is also not requiring boiling point testing. Vapor pressure was determined 

using thermogravimetric analysis following OECD guideline 104 appropriately. Two tests were 

performed to confirm the data collected and the measurement was given in both mm Hg and Pa 

units. EPA finds these results acceptable and would, therefore, not require vapor pressure 

testing.

Water solubility was visually determined by stirring for 48 hours and for 115 hours in 

double distilled water in a flask, which the study investigators say follows OECD guideline 

105. They mention visually determining the solubility but not any other more accurate 

analytical method. It is not clear how the study could adequately determine water solubility 

without using some analytical piece of equipment. EPA does not find this study acceptable as 

described, and if 1,4-Benzenedicarboxylic acid, 1,4-dimethyl ester, manuf. of, by-products 
50

from, met the finding of “substantial exposure,” EPA would still require that this testing be 

conducted. 

INVISTA also provided data for the log Kow endpoint. As described, the study was 

conducted using the HPLC method which is acceptable, however, the investigators ran the 

HPLC in a methanol/water mix. They also ran the experiment using known standards in the 

same methanol/water mixture. EPA considers, for this method to provide good data, known 

standards of similar structure must be used in order to calculate the log Kow of the sample via 

linear regression. The standards used are not that similar to the test sample in that none are 

benzoic acids or esters. If this testing were to be conducted, EPA believes this test should be 

rerun keeping the solvent system and standards in mind and is requiring log Kow testing in the 

final rule. 

The proposed test rule also included that ready biodegradation be determined. The 

ready biodegradability study submitted by INVISTA was conducted using a standard method 

(OECD 301B, CO2 Evolution Test (Modified Sturm Test) under GLP. The study is acceptable 

and fulfills the ready biodegradability test endpoint. No further ready biodegradability testing is 

needed.

The proposed test rule requires aquatic toxicity testing of 1,4-benzenedicarboxylic acid, 

1,4-dimethyl ester, manuf. of, by-products from (CASRN 68988-22-7). According to Table 3 

of the proposed test rule, the test requirements are for acute toxicity to fish and aquatic 

invertebrates and toxicity to aquatic plants testing if the log Kow for the test substance is less 

than 4.2 (i.e., the test substance belongs to Test Group 1). For a test substance with a log Kow

≥4.2 (i.e., the test substance belongs to Test Group 2), chronic toxicity to Daphnia and toxicity 

to plants (algae) testing are the required tests. 

INVISTA submitted the full study reports to EPA on December 3, 2010. The report 

provided compositional information on CASRN 68988-22-7. According to the analysis 

presented on page 38 of the report (Appendix 1), approximately 66.6% of the substance has 

been characterized. Of the 21 known components, three major constituents have been 
51

identified: dimethyl 1,4-benzenedicarboxylate (CASRN 120-61-8; 17.72%), trimethyl [1,1’-

biphenyl]-2,4’,5-tricarboxylate (CASRN 32741-92-7; 11.68%) and 1,4-benzenedicarboxylic 

acid, (4-methoxycarbonylphenyl)methyl methyl ester (CASRN 55334-51-5; 18.19%). The 

remaining components consist of esters of aromatic carboxylic acids, including benzene 

carboxylates, biphenyl carboxylates and carboxylates of polycyclic aromatics such as fluorene 

and dibenzo[b,d]pyran. More than 33% of the mixture remains uncharacterized. 

The log Kow determination for the test substance was made via the HPLC method 

(OECD Guidelines for the Testing of Chemicals no. 117). Table 3 in the report indicates that, 

when analyzed at a neutral pH, 30 component peaks were detected in the test substance, of 

which 15 had a log Kow ≥ 4.2. These 15 components together comprised approximately 25% of 

the total mixture. The water solubility of the substance was characterized as low (< 0.983 

mg/L). INVISTA also reported that acute Daphnia testing of DMT residue demonstrated 

toxicity, with a 48h-EC50 determination of 3.9 mg/L. 

Since nearly 25% of the proposed test rule substance is comprised of chemicals having 

log Kow values ≥ 4.2, the substance should be tested according to the requirements for Test 

Group 2. This determination is further supported by a characterization of the substance as 

having low solubility in water, evidence of moderate toxicity in acute Daphnia testing and no 

compositional information on approximately one-third of the proposed test rule substance.

INVISTA provided the report of an algae toxicity study on DMT residue. According to 

the report, this study was conducted in accordance with several guidelines, including OECD 

Guideline 201 (alga, growth inhibition test) and an OECD guidance document on aquatic 

toxicity testing of difficult substances and mixtures. Due to the low solubility of the substance, 

testing was performed using a water soluble fraction; this resulted in a 72h –EC50 determination 

of 3.4 mg/L. The submitted study is considered to be adequate, thus satisfying the requirement 

for toxicity to plants (algae) testing under Test Group 2. In order to satisfy the other 

requirement of Test Group 2, chronic toxicity to Daphnia testing would need to be conducted 

on 1,4-benzenedicarboxylic acid, 1,4-dimethyl ester, manuf. of, by-products from. INVISTA 

submitted the report of an acute Daphnia study, but not a chronic Daphnia study. Therefore, if 
52

this substance had remained in the test rule, a chronic toxicity to Daphnia test would still be 

required.

The proposed test rule also requires an acute mammalian toxicity test. INVISTA 

provided full studies for acute oral (OECD guideline 423) and dermal (OECD guideline 402) 

toxicity testing in rats of the proposed test rule substance. In the acute oral test, an LD50 > 2000 

mg/kg was reported. This is based on a limit test performed at 2000 mg/kg in which a single 

female rat (out of a total of 6 tested among two separate tests) was sacrificed moribund on day 

2 following administration of test material. In the acute dermal test, an LD50 > 2000 mg/kg was 

reported, based on 24-hour dermal application of test material at 2000 mg/kg and the survival 

of all treated rats. EPA considers the results to adequately characterize the hazard of acute oral 

and dermal exposure to CASRN 68988-22-7. Therefore, the proposed test rule requirement for 

acute mammalian toxicity testing is considered satisfied.

The proposed test rule also requires a bacterial reverse mutation assay and in vitro or in 

vivo testing for chromosomal damage. INVISTA submitted full study reports on genetic 

toxicity testing of CASRN 68988-22-7. Test results from a bacterial reverse mutation assay, a 

mammalian cell gene mutation assay (mouse lymphoma L5178Y cells), and an in vitro

chromosome aberration test of human peripheral blood lymphocytes were negative with and 

without exogenous metabolic activation. EPA considers the proposed genotoxicity test rule 

requirements to be satisfied.

The proposed test rule also requires repeated dose/reproduction/developmental 

mammalian toxicity testing. No information was provided on repeated 

dose/reproduction/developmental mammalian toxicity testing. These are required endpoints for 

testing and this requirement is not predicated on the degree of potential human exposure. 

In summary, if 1,4-Benzenedicarboxylic acid, 1,4-dimethyl ester, manuf. of, byproducts from, had met the finding of “substantial exposure,” EPA would still require water 

solubility, log Kow, and chronic toxicity to Daphnia. In addition, repeated

dose/reproduction/developmental mammalian toxicity would also still be required. However, 
53

the other test endpoints listed in the proposed rule for this chemical substance have adequate 

data and would not have been included in a final rule. 

12. 1-Decene, sulfurized (CASRN 72162-15-3).

Comment. AWOs stated that the proposed test rule requires fish and mammalian acute toxicity 

and combined repeated-dose reproduction/developmental toxicity screening tests for 1-decene, 

sulfurized, which has already been evaluated as part of the Alkyl Sulfide Category. AWOs 

noted that it became aware through a personal communication from an American Chemistry 

Council (ACC) employee that this chemical was dropped from the category in the final test 

plan revision for the Alkyl Sulfide Category because the sponsoring members discontinued 

manufacture and import of this chemical. AWOs stated that EPA should verify the current HPV 

status of this chemical. AWOs noted that EPA found the original Alkyl Sulfide Category test 

plan (which included 1-decene, sulfurized) to be “minimally acceptable,” that EPA believed 

these chemicals were unlikely to pose acute aquatic hazards due to their hydrophobicity, and 

that existing mammalian acute toxicity data indicated these compounds possess low acute 

toxicity hazard by a variety of exposure routes. AWOs consider the proposed fish and 

mammalian acute toxicity tests for 1-decene, sulfurized (CASRN 72162-15-3) to be 

unnecessary.

Response. The particular tests for aquatic toxicity testing of 1-decene, sulfurized (CASRN 

72162-15-3) in the proposed test rule depend on the log Kow for the chemical. For log Kow < 

4.2, testing is required for acute toxicity to fish and daphnia and toxicity to aquatic plants. For 

log Kow ≥ 4.2, testing is required for chronic toxicity to Daphnia and toxicity to aquatic plants.

EPA has reviewed the HPV file for the “alkyl sulfide” category and noted that the 

original test plan submitted by the CMA Association Petroleum Additives Panel Health, 

Environmental and Regulatory Task Group (HERTG) March 28, 2000, comprised five 

chemicals, including 1-decene, sulfurized. The test plan included no data for 1-decene, 

sulfurized regarding aquatic or mammalian toxicity endpoints.
54

HERTG provided results of studies for methyl propene derivative (CASRN 68511-50-2) 

that assessed its acute toxicity to fish, acute toxicity to invertebrates, and toxicity to aquatic 

plants using water accommodated fraction or water soluble fraction methodology; these 

endpoints were considered satisfied for 1-decene, sulfurized based on read across. In comments 

on the original test plan submission for the proposed alkyl sulfides category, EPA stated that 

the aquatic toxicity studies for methyl propene derivative were inadequate because they were 

not performed according to the most appropriate procedures for chemicals such as the alkyl 

sulfides that exhibit low water solubility and that testing should be performed at or below the 

water solubility limit. EPA noted that such substances are not likely to pose acute aquatic 

hazards, but may have potential to pose chronic aquatic hazards. EPA also suggested that 

aquatic toxicity testing requirements could be satisfied with a 21-day chronic daphnid test and a 

toxicity to aquatic plants test. EPA noted that chronic aquatic testing may not be necessary if 

the water solubility of the members of the proposed alkyl sulfides category (including 1-

decene, sulfurized) are sufficiently low. EPA maintains this position for the proposed test rule 

that requires aquatic toxicity testing for 1-decene, sulfurized.

HERTG provided results of mammalian acute oral toxicity studies for methyl propene 

derivative and propanol/dodecylthio derivative (CASRN 67124-09-8); acute inhalation toxicity 

studies for methyl propene derivative and trimethyl pentane derivative (CASRN 68515-88-8); 

and acute dermal toxicity studies for propanol/dodecylthio derivative, trimethyl pentane 

derivative, and C15-C18 alkene derivative (CASRN 67762-55-4); the mammalian acute 

toxicity endpoint was considered satisfied for 1-decene, sulfurized based on read across. EPA 

commented that the acute toxicity data for the four proposed alkyl sulfides category members 

did not show qualitative or quantitative differences and that the results suggested low acute 

toxicity, but also noted that the sponsor did not provide information regarding the mammalian 

acute toxicity endpoint for 1-decene, sulfurized, and that the biological responses for the other 

four proposed alkyl sulfides category members appear too low to rank the proposed category 

members according to biological activity.

HERTG provided results of studies of propanol/dodecylthio derivative (28-day oral study 

in rats), methyl propene derivative (13-week dermal study in rats and 21- and 28-day dermal 
55

studies in rabbits), and trimethyl pentane derivative (28-day inhalation and dermal studies in 

rats); the repeated-dose toxicity endpoint for 1-decene, sulfurized was considered satisfied 

based on read across. EPA was not convinced of the adequacy of a category approach for the 

repeated-dose endpoint because different routes of exposure were used across the studies and 

there was some suggestion of differences in toxicity among the substances tested. Therefore, 

EPA maintains the proposed test rule requirement for repeated-dose testing of 1-decene, 

sulfurized. The specific requirement is for a combined repeated-dose and 

reproductive/developmental toxicity screening test.

HERTG proposed to perform a one-generation reproduction study of 

propanol/dodecylthio derivative (intended to be applicable for the other alkyl sulfide category

members including 1-decene, sulfurized), and if results show low reproductive/developmental 

toxicity, no further testing would be considered necessary. EPA noted that a revised test plan 

for the proposed alkyl sulfides category, submitted by HERTG on May 18, 2005, does not 

include 1-decene, sulfurized. Therefore, in the absence of information to assess the 

reproductive/developmental toxicity of 1-decene, sulfurized, EPA maintains its requirement for 

a combined repeated-dose and reproductive/developmental toxicity screening test as stated in 

the proposed test rule.
56

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