Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

Study Tirle 

SATELLITE PROCEDUXE 

GASOLINE DIPE VAPOR CONDENSATE 

RAT MICRONUCLEUS TEST 

TEST GUIDELINES: US EPA Micronucleus Assay 79.64, CFR Vol. 59, No. 

122,27 June 1994. 

US EPA (1998) Health Effects Test Guidelines; OPPTS 

870.5395 Mammalian Erythrocyte Micronucleus Test. 

AUTHOR: Christine E. Mason, B.Sc. (Hons.) 

SUBCONTRACTOR: Huntingdon Life Sciences Ltd., 

Eye Research Centre (ERC) 

Eye, Suffolk 

IP23 7Px 

ENGLAND. 

HUNTINGDON SCIENCES 

LTD (PRC) NO.: 00-6130 

HUNTINGDON LIFE SCIENCES 

LTD (ERC) STUDY NO.: APT1009 

SUBCONTRACTOR'S SPONSOR: Huntingdon Life Sciences 

Princeton Research Center (PRC) 

Mettlers Road 

East Millstone, NJ 08875-2360 

USA 

Page 1 of 90 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

CONTENTS 

Page 

COMPLIANCE WlTH GOOD LABORATORY PRACTICE STANDARDS ......................... 4 

ERC . QUALITY ASSURANCE STATEMENT ...................................................................... 5 

PRC . QUALITY ASSURANCE STATEMENT ...................................................................... 6 

RESPONSIBLE PERSONNEL AND SCIENTIFIC APPROVAL ............................................ 7 

SUMMARY ................................................................................................................................ 

INTRODUCTION ...................................................................................................................... 

EXPERIMENTAL PROCEDURE ............................................................................................. 

ASSESSMENT OF RESULTS .................................................................................................. 

MAINTENANCE OF RECORDS .............................................................................................. 

RESULTS ................................................................................................................................... 15 

CONCLUSION ........................................................................................................................... 15 

REFERENCES ........................................................................................................................... 16 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

CONTENTS . continued 

TABLES 

1 . Summary of results and statistical analysis ...................................................................... 18 

2 . Results for individual animals . 24 hour sampling time .................................................. 19 

APPENDICES 

1 . Historical negative and positive control values ............................................................... 21 

2 . Animal exposure and observations data ........................................................................... 23 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS 

The slide evaluation phase of the study described in this report was conducted in compliance with the 

following Good Laboratory Practice standards and I consider the data generated to be valid. 

The UK Good Laboratory Practice Regulations 1999 (Statutory Instrument No. 3 106). 

OECD Principles of Good Laboratory Practice (as revised in 1997), 

ENVMClCHEM(98) 17. 

EC Commission Directive 199911 1EC of 8 March 1999 (Official Journal No. L 7718). 

US EPA 79.60, CFR Vol. 59, No. 122,27 June 1994. 

No compliance is claimed for work presented in the Experimental Procedure - In-life phase 

or Appendix 2 of this report. 

Christine E. Mason, B.Sc. (Hons.), 

Principal Investigator, 

Huntingdon Life Sciences Ltd., ERC. 

Date 

I am claiming compliance for the whole study with the following exceptions: 

The identity, strength, purity and composition or other characteristics to define the positive control 

article has not been determined by the Testing Facility. The positive control article has been 

characterized as per the Certificate of Analysis on file with the Testing Facility. The stability of the 

positive control article has not been determined by the Testing Facility. Analyses to determine the 

uniformity (as applicable) or concentration of the positive control mixture were not performed by the 

Testing Facility. The stability of the positive control article mixture has not been determined by the 

Testing Facility. 

......................... -- -- 

Gary M. Hoffman, B.A., D.A.B.T., 

Study Director, 

Huntingdon Life Sciences Ltd., PRC. 

Date 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

ERC - QUALITY ASSURANCE STATEMENT 

The following inspection and audit have been carried out in relation to the slide evaluation phase of 

this study: 

Study Phase Date of Inspection Date of Reporting 

Process Based Inspection 

Slide scoring 3 July 2002 5 July 2002 

6 September 2002 6 September 2002 

Report Audit 

Process Based Inspection: At or about the time this phase of the study was in progress, inspections 

of routine and repetitive procedures employed on this type of study were carried out. The slide 

scoring inspection was conducted and reported to appropriate Company Management as indicated 

above. 

Report Audit: This appendix has been audited by the Quality Assurance Department. This audit was 

conducted and reported to the Principal Investigator and Company Management as indicated above. 

Study based inspections were not performed on this phase of the study. 

The methods, procedures and observations were found to be accurately described and the reported 

results of this appendix to reflect the raw data. 

. . -..- -.. - 

Neal Jolly, B.Sc., M.R.Q.A., 

Group Manager, 

Department of Quality Assurance, 

Huntingdon Life Sciences Ltd. 

Date 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

PRC - QUALITY ASSURANCE STATEMENT 

Listed below are the dates that ths study was inspected by the Quality Assurance Unit of 

Huntingdon Life Sciences, East Millstone, New Jersey, and the dates that findings were 

reported to the Study Director and Management. This report reflects the raw data as far as can 

be reasonably established. 

Date(s) of Reported to Study 

Type of Inspection Inspection Director and Management 

Date 

Quality Assurance Auditor 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

RESPONSIBLE PERSONNEL AND SCIENTIFIC APPROVAL 

Gary M. Hoffman, B. A., D. A.B .T., 

Study Director 

Department of Safety Assessment, PRC. 

Date 

Christine E. Mason, B.Sc. (Hons.), 

Principal Investigator 

Department of Genetic Toxicology, ERC 

Chris Davison, B.Sc., 

Statistician 

Department of Statistics, ERC 

Date 

Date 
Huntingdon Life Sciences (ERC) Report No: APT 009/023700 

SUMMARY 

This satellite micronucleus study was designed to assess the potential induction of micronuclei by 

Gasoline DIPE Vapor Condensate in bone marrow cells of the rat. Animals were exposed for four 

weeks (5 days per week) by inhalation administration of the test substance at exposure levels of 2000, 

10000 and 20000 mg/m3. 

The test substance and negative control were administered by inhalation. The negative control group 

received clean air. A positive control group was dosed on one occasion by intraperitoneal injection, 

with cyclophosphamide at 40 mgfkg bodyweight. 

Bone marrow smears were obtained from five male and five female animals in the negative control 

and each of the test substance groups 24 hours after the 2oth exposure and from the positive control 

group 24 hours after dosing. One smear from each animal was examined for the presence of 

micronuclei in 2000 immature erythrocytes. The proportion of immature erythrocytes was assessed 

by examination of at least 1000 erythrocytes from each animal. A record of the incidence of 

micronucleated mature erythrocytes was also kept. 

No statistically significant increases in the frequency of micronucleated immature erythrocytes and no 

substantial decrease in the proportion of immature erythrocytes were observed in rats treated with 

Gasoline DPE Vapor Condensate compared to negative control values (P>0.01 in each case). 

The positive control compound, Cyclophosphamide, produced large, highly significant increases in 

the frequency of micronucleated immature erythrocytes and a decrease in the proportion of immature 

erythrocytes (P<O.OO 1). 

It is concluded that Gasoline DPE Vapor Condensate did not show any evidence of causing 

chromosome damage or bone marrow cell toxicity when administered by inhalation exposure in this 

in vivo test procedure. 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

INTRODUCTION 

The purpose of this satellite micronucleus study was to assess the potential of Gasoline DlPE Vapor 

Condensate to induce mutagenic effects in rats following inhalation admmistration using an in vivo 

cytogenetic system (Boller and Schmid 1970, MacGregor et a1 1987, Mavournin et a1 1990). The 

inhalation route was selected for use in this test as the most likely route of human exposure. 

The procedures used were based on the recommendations of the following guidelines: 

US EPA Micronucleus Assay 79.64, CFR Vol. 59, No. 122,27 June 1994. 

US EPA (1998) Health Effects Test Guidelines; OPPTS 870.5395 Mammalian 

Erythrocyte Micronucleus Test. 

The bone marrow micronucleus test, originally developed by Matter and Schmid (1971), is a widely 

employed and internationally accepted short-term assay for identification of genotoxic effects 

(chromosome damage and aneuploidy) associated with mutagens and carcinogens (Mavournin et a1 

1990). This in vivo system allows consideration of various factors including pharmacokinetics, 

metabolism and DNA repair which cannot be accurately modelled in an in vitro system. Young adult 

rats are chosen for use because of the high rate of cell division in the bone marrow, because of the 

wealth of background data on this species, and because of their general suitability for toxicological 

investigations. 

In mitotic cells in which chromosomal breakage has been caused by the test substance or its 

metabolites, acentric fragments of the chromosomes do not separate at the anaphase stage of cell 

division. After telophase these fragments may not be included in the nuclei of the daughter cells and 

hence will form single or multiple micronuclei (Howell-Jolly bodies) in the cytoplasm of these cells. 

Micronuclei are seen in a wide variety of cells, but erythrocytes are chosen for examination since 

micronuclei are not obscured by the main nucleus and are therefore easily detected in this cell type 

(Boller and Schmid 1970). 

Micronucleated immature erythrocytes appear in the bone marrow approximately 24 hours after 

induction of chromosome damage. These immature erythrocytes can be differentiated by a variety of 

staining techniques which rely on their relatively high content of residual RNA. Using the Feulgen 

method, they stain blue while mature erythrocytes (whch contain little RNA) are counterstained orange. 

An increased incidence of micronucleated immature erythrocytes is indicative of recent exposure to a 

chromosome-damaging agent. A simultaneous marked increase in the incidence of micronucleated 

mature erythrocytes is not expected and may be indicative of micronucleus-like artifacts (Schmid 1976). 

Substances which interfere with the mitotic spindle apparatus will cause non-disjunction (unequal 

separation of the chromosomes at anaphase resulting in aneuploidy) or lagging chromosomes at 

anaphase which may not be incorporated into the daughter nuclei. These lagging chromosomes are 

not excluded from the erythroblast with the main nucleus and hence also give rise to micronuclei. 

Any toxic effects of the test substance on the nucleated cells may lead either to a reduction in cell 

division or to cell death. These effects in turn lead to a reduction in the number of nucleated cells 

and immature erythrocytes; to compensate for this, peripheral blood is shunted into the bone marrow 

(von Ledebur and Schmid 1973). If the proportion of immature erythrocytes is found to be 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

significantly less than the control value, this is taken as being indicative of toxicity. A very large 

decrease in the proportion would be indicative of a cytostatic or cytotoxic effect. 

The slide evaluation phase of the satellite micronucleus study was performed at the Department of 

Genetic Toxicology, Huntingdon Life Sciences (ERC), Eye, Suffolk, IP23 7PX, England. 

The experimental start and completion dates of the slide evaluation phase of the study were 7 May 

2002 and 30 July 2002 respectively. 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

EXPERTMENTAL PROCEDURE 

In-life phase 

The in-life phase of the study was carried out at the Princeton Research Center starting on 15 March 

2002 and was completed on 11 April 2002. 

All animals in the negative control and test substance groups were exposed for four weeks (5 days per 

week) by inhalation. The non-exposed positive control group was dosed with Cyclophosphamide 

administered on one occasion by intraperitoneal injection at a volume dosage of 10 mlkg 

bodyweight. Cyclophosphamide (CP, CAS # 6055-1 9-2, lot number 1 O8HO568, received 10 April 

2002, expiration 30 June 2002, white powder, storage 2-8"C, purity 99.2%), was obtained from the 

Sigma Chemical Company (responsible for its characterization), and was dissolved and diluted in 

sterile distilled water at Huntingdon Life Sciences to stock concentrations of 4.0 mglmL for use as 

the positive control for the micronucleus study. 

The experimental design is shown below: 

Group Treatment Exposure Level Animal Numbers 

(mg/m3) Male Female 

1 Air control 1081 - 1085 1591 - 1595 

2 Test Substance 2000 2071 - 2075 2581 - 2585 

3 Test Substance 10000 3071 - 3075 3581 - 3585 

4 Test Substance 20000 408 1 - 4085 4591 - 4595 

6 Cyclophosphamide 40 (mgkg) 6051 - 6055 6561 - 6565 

Five males and five females from the negative control and each of the test substance groups were 

sacrificed 24 hours after the final exposure period by isoflurane inhalatiodexsanguination. Five 

males and five females from the positive control group were sacrificed 24 hours after CP dosing by 

C02 inhalatiodexsanguination. Both femurs were exposed, cut just above the knee and the bone 

marrow was aspirated into a syringe containing a small volume (about 0.5 mL) of serum. The cells 

were then flushed into a centrifuge tube of cold serum. The tubes were identified by labels 

containing the study, group number, and animal number. 

The bone marrow cells were pelleted by centrifugation at about 150 x g for about 5 min and the 

supernatant drawn off, leaving a small amount of serum with the cell pellet. The cells were 

resuspended by aspiration with a pasteur pipette and a small drop of cells was spread onto a clean 

glass slide. Four slides were prepared from each animal. The slides were allowed to air dry, fixed by 

dipping for about 3 minutes in methanol, and aged overnight or longer prior to staining. Slides were 

labelled with experiment and animal number using a lead pencil. 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

Two slides from each animal were despatched to Huntingdon Life Sciences (ERC), Eye, Suffolk, 

IP23 7PX, England for slide staining and analysis. The remaining 2 smears and the cell pellet 

(refrigerated) were held in reserve at PRC in case of technical problems with the first 2 smears. 

Slide evaluation 

Due to the presence of mast cell granules in rat bone smears, which appear identical to micronuclei 

when stained using the Romanowslq methods, a modified Feulgen staining method is employed for 

the rat micronucleus test in this laboratory. This method specifically stains DNA-containing bodies 

deep purple while leaving mast cell granules unstained. The method also allows reasonable 

differentiation of mature and immature erythrocytes and produces permanent preparations. 

Two slides from each animal were stained as follows: 

1. Hydrolysed in Bouin's fluid at room temperature for approximately 30 hours. 

2. Washed three times in purified water (5 minutes per wash). 

3. Stained in Schiffs reagent for one hour at room temperature. 

4. Washed three times in purified water (5 minutes per wash). 

5. Counter-stained for ten minutes in very dilute (approximately 0.06 gll) aqueous Eosin 

yellowish. 

6. Washed for five minutes in purified water. 

7. Stained for 30 minutes in Mayer's Haemalum diluted 9 volumes: 1 volume with aqueous 

acridine orange solution in purified water (1 mgtrnl). 

8. Rinsed in purified water. 

9. Rinsed in running tap water. 

10. Washed for 5 minutes in purified water. 

11. Ax-dned. 

12. Slides were mounted with coverslips using DPX mountant. 

13. The mountant was allowed to harden at approximately 37OC. 

NB All stains and Bouin's fluid were filtered immediately prior to use to remove particulate material. 

The stained smears were examined (under code) by light microscopy to determine the incidence of 

micronucleated cells per 2000 polychromatic erythrocytes per animal. One smear per animal was 

examined. The remaining smears were held temporarily in reserve in case of technical problems with 

the first smear. 

Micronuclei are identified by the following criteria: 

Large enough to discern morphological characteristics 

Should possess a generally rounded shape with a clearly defined outline 

Should be deeply stained and similar in colour to the nuclei of other cells - not black 

Should lie in the same focal plane as the cell 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

Lack internal structure, ie they are pyknotic 

There should be no micronucleus-like debris in the area surrounding the cell 

The proportion of immature erythrocytes for each animal was assessed by examination of at least 

1000 erythrocytes. A record of the number of micronucleated mature erythrocytes observed during 

assessment of this proportion was also kept as recommended by Schd (1976). 

Deviations from Protocol 

This phase of the study was conducted in compliance with the following additional Good Laboratory 

Practice Standards: the UK Good Laboratory Practice Regulations 1999 (Statutory Instrument No. 

3106), the OECD Principles of Good Laboratory Practice (as revised in 1997), 

ENV/MC/CHEM(98)17 and the EC Commission Directive 199911 11EC of 8 March 1999 (Official 

Journal No. L 7718). 

The statistical analysis was performed at Huntingdon Life Sciences Ltd., Eye, Suffolk, P23 7PX, 

England. 

ASSESSMENT OF RESULTS 

The results for each treatment group were compared with the results for the concurrent negative 

control group using non-parametric statistics. Non-parametric statistical methods were chosen for 

analysis of results because: 

They are suited to analysis of data consisting of discretelinteger values with ties such 

as the incidence of micronucleated immature erythrocytes. 

The methods make few assumptions about the underlying distribution of data and 

therefore the values do not require transformation to fit a theoretical distribution 

(where data can be approximately fitted to a normal distribution, the results of nonparametric analysis and classical analysis of variance are very similar). 

'Outliers' are frequently found in the proportion of immature erythrocytes for both 

control and treated animals; non-parametric analysis based on rank does not give 

these values an undue weighting. 

For incidences of micronucleated immature erythrocytes, exact one-sided p-values are calculated by 

permutation (StatXact, CYTEL Sofhvare Corporation, Cambridge, Massachussetts). Comparison of 

several dose levels is made with the concurrent control using the Linear by Linear Association test 

for trend, in a step-down fashion if significance is detected (Agresti et al. 1990); for individual intergroup comparisons (ie the positive control group) this procedure simplifies to a straightforward 

permutation test (Gibbons 1985). For assessment of effects on the proportion of immature 

erythrocytes, equivalent permutation tests based on rank scores are used, ie exact versions of 

Wilcoxon's sum of ranks test and Jonckheere's test for trend. 
Huntingdon Life Sciences (ERC) Report No: APT 009/023700 

A positive response is normally indicated by a statistically significant dose-related increase in the 

incidence of micronucleated immature erythrocytes for thk treatment group compared with the 

concurrent control group (P<0.01); individual andlor group mean values should exceed the laboratory 

historical control range (Morrison and Ashby 1995). 

A negative result is indicated where individual and group mean incidences of micronucleated 

immature erythrocytes for the group treated with the test substance are not significantly greater than 

incidences for the concurrent control group (P>0.01) and where these values fall within the historical 

control range. An equivocal response is obtained when the results do not meet the criteria specified 

for a positive or negative response. 

Bone marrow cell toxicity (or depression) is normally indicated by a substantial and statistically 

significant dose-related decrease in the proportion of immature erythrocytes (P<0.01). 

MAINTENANCE OF RECORDS 

All raw data, samples and specimens arising from the performance of this phase of the study will 

remain the property of the Sponsor. 

Types of sample and specimen that are unsuitable, by reason of instability, for long term retention 

and archiving may be disposed of after the periods stated in Huntingdon Life Sciences, Standard 

Operating Procedures. 

All other samples and specimens and all raw data will be retained by Huntingdon Life Sciences PRC 

in its archive for a period of one year from the date on which the Study Director signs the final report. 

After such time, the Sponsor will be contacted and their advice sought on the return, disposal or 

further retention of the materials. If requested, Huntingdon Life Sciences will continue to retain the 

materials subject to a reasonable fee being agreed with the Sponsor. 

Huntingdon Life Sciences will retain the Quality Assurance records relevant to this study and a copy 

of the final report in its archive indefinitely. 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

RESULTS 

MICRONUCLEUS TEST 

Micronucleated immature erythrocyte counts (mie) 

The test substance did not cause any statistically significant increases in the number of 

micronucleated immature erythrocytes [P>0.0 11. 

Cyclophosphamide caused large, highly significant increases in the frequency of micronucleated 

immature erythrocytes [P<0.00 11. 

Micronucleated mature erythrocytes (mme) 

The test substance did not cause any substantial increases in the incidence of micronucleated mature 

erythrocytes. 

Proportion of immature erythrocytes (% ie/[ie + me]) 

The test substance failed to cause any significant decreases in the proportion of immature 

erythrocytes [P>0.0 11. 

Cyclophosphamide caused statistically significant decreases in the proportion [P<0.001]. 

CONCLUSION 

No statistically significant increases in the frequency of micronucleated immature erythrocytes and 

no substantial decrease in the proportion of immature erythrocytes were observed in rats treated with 

Gasoline DIPE Vapor Condensate compared to negative control values (P>0.01 in each case). 

It is concluded that Gasoline DIPE Vapor Condensate did not show any evidence of causing 

chromosome damage or bone marrow cell toxicity when administered by inhalation in this in vivo test 

procedure. 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

REFERENCES 

AGRESTI, A., MEHTA, C.R. and PATEL, N.R. (1990) Exact inference for contingency tables with 

ordered categories. Journal of the American Statistical Association, 85,453. 

BOLLER, K. and SCHMID, W. (1970) Chemical mutagenesis in mammals. The bone marrow of the 

Chinese hamster as an in vivo test system. Haematological findings after treatment with Trenimon 

(translation). Humangenetik, 11, 34. 

CYTEL (1995) StatXact 3 for Windows: Statistical Software for Exact Nonparametric Inference. Cytel 

Software Corporation, NC, USA. 

GIBBONS, J.D. (1985) Nonparametric Statistical Inference, 2nd edition, Marcel Dekker, New York. 

JONCKHEERE, A.R. (1954) A distribution-fi-ee k-sample test against ordered alternatives. 

Biometries, 41, 133-145. 

KRUSKAL, W.H. and WALLIS, W.A. (1952) Use of Ranks in One-Criterion Variance Analysis. 

Journal of the American Statistical Association, 47,5 83-62 1. 

KRUSKAL, W.H. and WALLIS, W.A. (1953) Errata for Kruskal-Wallis (1952). Journal of the 

American Statistical Association, 47,583-62 1. 

MacGREGOR, J.T., HEDDLE, J.A., HITE, M., MARGOLIN, B.H., RAMEL, C., SALAMONE, 

M.F., TICE, R.R. and WILD, D. (1987) Guidelines for the conduct of micronucleus assays in 

mammalian bone marrow erythrocytes. Mutation Research, 189, 103. 

MATTER, B. and SCHMID, W. (1971) Trenimon-induced chromosomal damage in bone marrow 

cells of six mammalian species, evaluated by the micronucleus test. Mutation Research, 12,417. 

MAVOURNIN, K.H., BLAKEY, D.H., CIMINO, M.C., SALAMONE, M.F. and HEDDLE, J.A. 

(1990) The in vivo micronucleus assay in mammalian bone marrow and peripheral blood. A report of 

the US Environmental Protection Agency Gene-Tox Program. Mutation Research, 239,29. 

MORRISON, V. and ASHBY, J. (1995) High resolution rodent bone marrow micronucleus assays of 

1,2-dimethylhydrazine : implication of systemic toxicity and individual responders. Mutagenesis, 10, 

129. 

SAS INSTITUTE (1989) SAS/STAT User's Guide, Version 6, Fourth Edition, Vo1.2. SAS Institute Inc., 

Cary, NC, USA. 

SAS INSTITUTE (1 996) SAS/STAT Software: Changes and Enhancements through Release 6.1 1. SAS 

Institute, Cary, NC, USA. 

SAS INSTITUTE (1996) SAS/STAT Software: Changes and Enhancements for Release 6.12. SAS 

Institute, Cary, NC, USA. 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

SCHMID, W. (1976) The micronucleus test for cytogenetic analysis. In: HOLLANDER, A. (ed.) 

Chemical Mutagens, Principles and Methods for their Detection, 4, 3 1. Published by Plenum Press, 

New York. 

von LEDEBUR, M. and SCHMID, W. (1973) The rnicronucleus test. Methodological aspects. 

Mutation Research, 19, 109. 

WILCOXON, F. (1945). Individual comparisons by ranking methods. Biometries Bulletin, 1, 80-83. 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

TABLE 1 

Summary of results and statistical analysis 

Sampling Treatment Exposure level % iel(ie+me) t Incidence mie Incidence rnme 

time (mg/m" (group mean) (group mean) (group mean)" 

24 Hours Negative control 5 1 0.8 0.0 

TS 2000 52 0.8 1.1 

TS 10000 49 0.9 1.1 

TS 20000 53 0.7 0.8 

Cyclophosphamide 40 mgkg 43*** 15.1""" 0.0 

TS Gasoline DIPE Vapor Condensate 

% ie/(ie+me) Proportion of immature erythrocytes 

mie Number of micronucleated cells observed per 2000 immature erythrocytes examined 

mme Number of micronucleated cells calculated per 2000 mature erythrocytes 

Results of statistical analysis using the appropriate nonparametric method of analysis based on 

permutation (one-sided probabilities): 

*** P < 0.001 (highly significant) 

otherwise P > 0.01 (not significant) 

1- Occasional apparent errors of k 1% may occur due to rounding of values for presentation in the 

table 

" Formula for calculation of incidence mme (group mean): 

Sum of mouv incidence mme scored x 2000 

Sum of group me scored 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

TABLE 2 

Results for individual animals - 24 hour sampling time 

I Treatment Exposure level Animal ie me % ie/(ie+me) Incidence Incidence 

(mg/m') number mie me 

Negative control - M 1081 486 594 45 0 0 

M 1082 593 527 5 3 1 

M 1083 650 612 52 1 

M 1084 556 515 5 2 1 

M 1085 626 522 5 5 1 

F 1591 567 53 1 52 2 

F 1592 567 503 53 0 

F 1593 519 594 47 1 

F 1594 542 507 52 1 

F 1595 564 555 5 0 0 

TS 2000 M 2071 532 . 520 5 1 0 

M 2072 572 490 54 0 

M 2073 64 1 547 54 2 

M 2074 582 571 50 1 

M 2075 600 505 54 2 

F 2581 603 5 64 52 0 

F 2582 524 517 50 1 

F 2583 620 566 52 0 

F 2584 509 524 49 1 

F 2585 560 500 5 3 1 

TS 10000 M 3071 531 . 536 5 0 1 

M 3072 530 510 5 1 1 

M 3073 491 585 46 0 

M 3074 582 593 5 0 1 

M 3075 516 568 48 1 

F 3581 485 599 45 1 

F3582 498 603 45 2 

F 3583 574 479 5 5 1 

F3584 519 536 49 1 

F 3585 626 603 5 1 0 

TS 20000 M 4081 63 5 65 1 49 1 

M4082 580 457 56 1 

M 4083 642 489 57 1 

M 4084 545 535 5 0 0 

M 4085 529 50 1 5 1 0 

F 4591 616 43 1 5 9 1 

F4592 582 555 5 1 1 

F 4593 665 427 61 0 

F4594 518 595 47 1 

F 4595 524 574 48 1 

TS Gasoline DPE Vapor Condensate 

% iel(ie+me) Proportion of immature erythrocytes 

mie Number of micronucleated cells observed per 2000 immature erythrocytes 

me Total number of mature erythrocytes examined for micronuclei 

me Number of micronucleated mature erythrocytes observed 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

TABLE 2 - continued 

Results for individual animals - 24 hour sampling time 

Treatment Dosage Animal ie me % iel(ie+me) Incidence Incidence 

number mie mme 

C~clophosphamide 40 mgkg M 605 1 422 604 4 1 13 0 

M 6052 525 593 47 11 0 

M 6053 506 63 1 45 7 0 

M 6054 503 694 42 12 0 

M 6055 487 55 1 47 6 0 

F6561 419 609 4 1 19 0 

F 6562 483 623 44 25 0 

F 6563 462 599 44 25 0 

F 6564 422 683 3 8 9 0 

F 6565 473 72 1 40 24 0 

% ie/(ie+me) Proportion of immature 

mie Number of micronucleated cells observed per 2000 immature erythrocytes 

me Total number of mature erythrocytes examined for micronuclei 

mme Number of micronucleated mature erythrocytes observed 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

APPENDIX 1 

Historical negative control values (1 July2000 - 30 June 2002) 

Frequency of micronucleated immature erythrocytes (individual animals) 

Number of micronucleated immature erythrocytes 

(MIE per 2000) 

Individual man 0.79 

Historical negative controlvalues (1 July2000 - 30 June 2002) 

Frequency of micronucleated immature erythrocytes (Group mean values) 

Number of micronucleated immature erythrocytes (MIEper 2000) 

Group man 0.73 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

APPENDLX 1 - continued 

Historical positive control values (1 July 2000 - 30 June 2002) 

Frequency of micronucleated immature erythrocytes (Individual animals) 

Cyclophosphamide 

Number of micronucleated immature erythrocytes 

(M lE per 2000 cells) 

Individual mean 38.7 

Historical positive control values (1 July 2000 - 30 June 2002) 

Frequency of micronucleated immature erythrocytes (Group mean values) 

Cyclophosphamide 

Number of micronucleated immature erythrocytes (MIEper 2000) 

Group mean 39.9 
Huntingdon Life Sciences (ERC) Report No: APT 0091023700 

APPENDIX2 . 

ANIMAL EXPOSURE AND OBSERVATIONS DATA 
Huntingdon Life Sciences 00-6130 Page 1 

2 1 1 -DIPE-S Draft Final Report 

Animal Exposure and Animal Data 

Preface I Appendix 2 1 

INTRODUCTION: The following is data generated at Huntingdon Life Sciences, East 

Millstone, NJ. The separately issued main study report should be referenced for details of 

the procedures used for test atmosphere generationkharacterization and animal 

evaluations. 

STUDY DATES: Date of Animal Receipt: 4 March 2002 

Experimental Initiation Date: 15 March 2002 (in-life) 

Experimental Completion Date: 11 April 2002 (in-life) 

Draft Report Date: 6 March 2003 

EXPOSURES AND IN-LIFE SUMMARY: The actual measured results during the 

exposures were comparable to the targeted exposure levels. There were no exposurerelated effects seen in the test animals with regards to body weights and feed 

consumption. 

TABLE OF CONTENTS 

TABLES 

A. Chamber Monitoring Results ........................................................................................... 

B. Summary of Clinical Observations .................................................................................. 

C. Mean Body Weights (grams) ........................................................................................... 

D. Mean Body Weight Change (grams) ................................................................................ 

E. Mean Feed Consumption Values (gramslkglday) ............................................................ 

F. Individual Clinical Observations ..................................................................................... 

G. Individual Body Weights (grams) .................................................................................... 

H. Individual Body Weight Change (grams) ........................................................................ 

I. Individual Feed Consumption Values (grams/kg/day) ..................................................... 

J. Animal Termination History ............................................................................................ 
Table A GASOLINE DlPE VAPOR CONDENSATE: A 13-WEEK 00-61 30 

WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 

Date Exposure 

Number 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 

S.D. 

Chamber Monitoring Results 

Cumulative Exposure Record 

Group IA - 0 mg/m3 (Air Control) 

Nominal 

(w/m3) 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

Mean 

(mglm3) 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

alytical Chamber Concentration 

Individual 

(ms 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

MMAD 

(crm) 

0.9053 

2.495 

2.357 

2.187 

1.986 

0.731 

Particle Size 

:terminat 

GSD 

1s 

TMC 

(mg/m3) 

1.20E-02 

4.91 E-03 

7.60E-03 

2.32E-03 

6.71 E-03 

4.13E-03 

Chamber Environment 

h, 

Temperature 

("C) 

25 

27 

25 

24 

24 

24 

24 

24 

25 

25 

25 

25 

25 

24 

23 

24 

23 

24 

25 

25 

24.5 

0.9 

In 

Humidity 

(%I 

53 

54 

53 

54 

52 

53 

58 

53 

5 1 

52 

55 

54 

50 

56 

52 

55 

55 

51 

51 

53 

53.3 

1.9 
Table A GASOLINE DlPE VAPOR CONDENSATE: A 13-WEEK 

WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 

Chamber Monitoring Results 

Cumulative Exposure Record 
Table A GASOLINE DlPE VAPOR CONDENSATE: A 13-WEEK 00-61 30 

WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 

Chamber Monitoring Results 

Cumulative Exposure Record 

Group IIA - 2,000 mglrn3 

Chamber Environment 

Particle Size Mean 

Analytical Chamber Concentration 

- - 

- 

Humidity 

("/.I 

50 

50 

48 

50 

49 

50 

54 

51 

48 

48 

52 

50 

47 

52 

48 

5 1 

51 

48 

47 

49 

49.7 

1.8 

1s 

TMC 

(rnglm3) 

1 .I 9E-02 

4.72E-03 

4.56E-03 

1.53E-03 

5.68E-03 

4.40E-03 

remperature 

("'3 

23 

26 

23 

23 

23 

23 

23 

23 

24 

24 

24 

24 

24 

23 

23 

23 

23 

23 

24 

24 

23.5 

0.8 

Date Exposure 

Number 

Nominal 

(mg/m3) 

21 60 

2190 

2190 

21 60 

21 30 

2180 

2050 

2090 

2030 

2170 

2110 

2110 

2090 

2030 

2130 

2060 

2220 

1980 

2040 

2050 

2109 

67 

MMAD 

(clrn) 

0.794 

3.91 1 

0.8422 

1.438 

1.746 

1.473 

Mean 

(mglm3) 

I968 

2068 

2205 

2095 

2040 

21 08 

1913 

1918 

1913 

21 33 

2090 

2028 

I843 

I845 

I875 

2093 

2150 

1950 

I938 

1955 

Individual 

(ms 

1920 

2050 

21 90 

2060 

2030 

2040 

1940 

1900 

1760 

21 30 

2030 

2040 

1630 

1790 

1890 

21 30 

2030 

1810 

2030 

1950 

2006 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 

S.D. 
Table A GASOLINE DlPE VAPOR CONDENSATE: A 13-WEEK 00-61 30 

WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 

Date ixposure 

Number 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 

S.D. 

Nominal 

(mg/m3) 

2160 

21 90 

2190 

2160 

21 30 

21 80 

2050 

2090 

2030 

2170 

2110 

2110 

2090 

2030 

21 30 

2060 

2220 

1980 

2040 

2050 

21 09 

67 

Chamber Monitoring Results 

Cumulative Exposure Record 

Grow IIB - 2.000 mdm3 

Mean 

(mg/m3) 

2020 

2098 

2020 

2028 

2040 

2073 

1960 

I968 

1960 

I978 

1915 

1998 

21 30 

1960 

1970 

2098 

2055 

I858 

1868 

1943 

alytical Chamber Concentration 

Individual 

(ms 

2020 

2200 

201 0 

1930 

2060 

2020 

1990 

2050 

2000 

1980 

1920 

1980 

2400 

1840 

2020 

2220 

1890 

1790 

1780 

1980 

1997 

MMAD 

(vm) 

0.8998 

0.9819 

0.8897 

1.403 

1.044 

0.243 

Particle Size 

Determinations 

GSD TMC 

(mg/m3) 

1.47E-02 

1.76E-03 

5.04E-03 

1.74E-03 

5.81 E-03 

6.1 3E-03 

n 

Humidity 

(%) 

52 

52 

50 

52 

51 

51 

55 

52 

52 

50 

54 

53 

48 

54 

49 

54 

54 

49 

52 

52 

51.8 

1.9 
Table A GASOLINE DlPE VAPOR CONDENSATE: A 13-WEEK 00-61 30 

WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 

Date Exposure 

Number 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 

S.D. 

Chamber Monitoring Results 

Cumulative Exposure Record 

Group lllA - 10,000 mg/m3 

Nominal 

(mg/m3) 

9980 

9530 

10000 

10200 

10100 

10300 

9370 

9690 

9940 

9390 

9540 

9670 

9830 

10000 

10600 

10000 

9470 

10000 

9880 

10100 

9880 

32 1 

1 

Mean 

(mglm3) 

9973 

lo260 

10000 

10320 

1001 0 

9970 

9475 

9450 

9773 

9670 

9930 

1001 0 

9790 

9970 

9730 

9745 

9098 

10450 

9828 

9945 

~alytical Chamber Concentration 

Individual 

Particle Size 

MMAD 

(IJm) 

0.7927 

1.316 

0.7720 

1.490 

1 .O93 

0.365 

sterminat 

GSD 

1s 

TMC 

(mglm3) 

2.41 E-02 

2.91 E-03 

5.54E-03 

1.78E-03 

8.58E-03 

1.05E-02 

Chamber 

M 

remperature 

("'7 

23 

26 

24 

23 

23 

23 

23 

23 

24 

24 

24 

24 

24 

23 

23 

23 

23 

23 

24 

24 

23.6 

0.8 

vironment 

n 

Humidity 

(%) 

50 

50 

47 

50 

50 

49 

50 

48 

48 

48 

48 

49 

47 

50 

48 

48 

49 

47 

49 

49 

48.7 

1 .I 
Table A GASOLINE DlPE VAPOR CONDENSATE: A 13-WEEK 00-61 30 

WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 

Chamber Monitoring Results 

Date xposure 

Number 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 

S.D. 

Nominal 

(m!3/m3) 

9980 

9530 

10000 

10200 

101 00 

10300 

9370 

9690 

9940 

9390 

9540 

9670 

9830 

10000 

10600 

10000 

9470 

10000 

9880 

10100 

9880 

321 

Cumulative Exposure Record 

Group lllB -10.000 mg/rn3 

Chamber Environment 

Mean 

(mg1rn3) 

9983 

9325 

9980 

9548 

951 0 

10110 

9188 

9573 

10430 

9818 

9493 

10070 

lo260 

9033 

lo630 

10680 

10250 

9763 

10130 

lo780 

alytical Chamber Concentration 

Individual MMAD 

(IJrn) 

0.7866 

4.191 

0.7259 

1.767 

1.868 

1.621 

Particle Size 

Determinations 

GSD TMC 

(mg1m3) 

2.44E-02 

5.14E-03 

5.49E-03 

3.29E-03 

9.58E-03 

9.93E-03 

Temperature 

("C) 

23 

26 

23 

24 

24 

24 

24 

24 

23 

23 

23 

24 

24 

24 

24 

24 

24 

24 

24 

23 

23.8 

0.7 

In 

Humidity 

("/.I 

49 

49 

46 

46 

46 

46 

46 

45 

46 

46 

46 

47 

44 

47 

46 

45 

45 

45 

48 

46 

46.2 

1.3 
Table A GASOLINE DlPE VAPOR CONDENSATE: A 13-WEEK 00-61 30 

WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 

Chamber Monitoring Results 

Cumulative Exposure Record 

Group IVA - 20,000 mglm3 

Date Exposure 

Number 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 

S.D. 

Nominal ~alytical Chamber Concentration 

(mglm3) 

191 00 

19400 

I9000 

I9300 

19800 

I9000 

I9500 

I9300 

19300 

19600 

18400 

18600 

18700 

18900 

18800 

18900 

191 00 

18800 

I7600 

19700 

19040 

504 

Individual 

1 

Mean 

(mglm3) 

19100 

19980 

19880 

19250 

19650 

19350 

20280 

19550 

20330 

20250 

19780 

19480 

20080 

20450 

20300 

19480 

20100 

19830 

I9050 

20780 

(mo 

I8500 

20500 

20300 

19000 

20400 

19500 

20300 

1 9700 

20100 

1 9700 

19800 

18900 

19700 

20200 

19800 

19000 

19500 

19500 

18800 

20800 

19850 

MMAD 

(~.rm) 

0.7739 

2.346 

0.8053 

1.220 

1.286 

0.735 

Particle Size 

Determinations 

GSD TMC 

(m91m3) 

2.44E-02 

4.30E-03 

7.67E-03 

1.09E-03 

9.37E-03 

1.04E-02 

Chamber Environment 

rn 

Humidity 

("A) 

52 

52 

48 

51 

50 

50 

51 

49 

49 

49 

48 

49 

47 

50 

52 

50 

50 

49 

50 

51 

49.9 

1.4 
Table A GASOLINE DlPE VAPOR CONDENSATE: A 13-WEEK 00-61 30 

WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 

Chamber Monitoring Results 

Cumulative Exposure Record 

Group IVB - 20,000 mglm3 

Chamber Environment 

Particle Size 

Determinations 

M 

Temperature 

("C) 

24 

26 

24 

25 

25 

25 

25 

25 

24 

24 

24 

24 

24 

25 

25 

25 

25 

25 

25 

24 

24.7 

0.6 

In 

Humidity 

(%I 

51 

52 

49 

50 

49 

49 

50 

48 

5 1 

50 

50 

50 

48 

49 

50 

49 

48 

48 

52 

50 

49.7 

1.2 

Nominal 

(mglm3) 

19100 

19400 

19000 

19300 

19800 

19000 

19500 

19300 

19300 

19600 

I8400 

18600 

18700 

I8900 

18800 

18900 

191 00 

18800 

I7600 

19700 

19040 

504 

Date Exposure 

Number 

&tical Chamber Concentration 

Individual TMC 

(mglm3) 

2.31 E-02 

3.14E-03 

9.04E-03 

2.38E-03 

9.42E-03 

9.60E-03 

MMAD 

(~m) 

0.7578 

0.9018 

0.7779 

1.517 

0.989 

0.358 

Mean GSD 

(mglm3) 

20430 

20680 

20980 

20580 

20800 

20730 

20750 

21 030 

19850 

20380 

19930 

19650 

18930 

18830 

19250 

19900 

19700 

19880 

18230 

20350 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 1 

TABLE B 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

NORMAL 

WITHIN NORMAL LIMITS 

DEAD 

TERMINAL SACRIFICE 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 2 

TABLE B 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

FEMALES SUMMARY OF CLINICAL OBSERVATIONS 

# OF ANIMALS EXAMINED 155 

255 

355 

4 5 5 

NORMAL 

WITHIN NORMAL LIMITS 

DEAD 

TERMINAL SACRIFICE 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 1 

TABLE C 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

WEEK 

WEEK 

WEEK 

WEEK 

WEEK 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N ........................................... 

No statistically significant differences 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 2 

TABLE C 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

WEEK 0 

WEEK 1 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

WEEK 2 

WEEK 3 MEAN 

S.D. 

N 

WEEK 4 MEAN 

S.D. 

N -------------- 

differences 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 1 

TABLE D 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

WEEK 0 TO 1 MEAN 

S.D. 

N 

WEEK 0 TO 2 MEAN 

S.D. 

N 

WEEK 0 TO 3 MEAN 

S.D. 

N 

No statistically significant differences 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 2 

TABLE D 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

WEEK 0 TO 1 

WEEK 0 TO 2 

WEEK 0 TO 3 

WEEK 0 TO 4 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

MEAN 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 1 

TABLE E 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MALES MEAN FEED CONSUMPTION VALUES (GRAMS/KG/DAY) ................................................................................................................................... 

DOSE GROUP: 1 2 3 4 6 

DOSE LEVEL (MG/M3) : 0 2000 10000 20000 MICRO+CONTROL ................................................................................................................................... 

WEEK 1 MEAN 

S.D. 

N 

WEEK 2 MEAN 

S.D. 

N 

WEEK 3 MEAN 

S.D. 

N 

WEEK 4 MEAN 

S.D. 

N ............................................... 

No statistically significant differences 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 2 

TABLE E 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

FEMALES MEAN FEED CONSUMPTION VALUES (GRAMS/KG/DAY) ................................................................................................................................... 

DOSE GROUP: 1 2 3 4 6 

DOSE LEVEL (MG/M3): 0 2000 10000 20000 MICRO+CONTROL ................................................................................................................................... 

WEEK 1 MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

WEEK 2 

WEEK 3 MEAN 

S.D. 

N 

WEEK 4 MEAN 

S.D. 

N 

---------------- 

Statistical key: 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 1 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

1081 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

1082 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

1083 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

1084 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

1085 WITHIN NORMAL LIMITS P 

TERMINAL SACRIFICE P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 2 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL CLINICAL OBSERVATIONS 

MALES GROUP 2 2000 MG/M3 

2071 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

2072 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

2073 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

2074 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

2075 WITHIN NORMAL LIMITS P 

TERMINAL SACRIFICE P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 3 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL CLINICAL OBSERVATIONS 

MALES GROUP 3 10000 MG/M3 ................................................................................................................................... 

3071 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

3072 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

3073 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

3074 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

3075 WITHIN NORMAL LIMITS P 

TERMINAL SACRIFICE P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 4 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

4081 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

4082 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

4083 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

4084 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

4085 WITHIN NORMAL LIMITS P 

TERMINAL SACRIFICE P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 5 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

6051 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

6052 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

6053 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

6054 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

6055 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 6 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL CLINICAL OBSERVATIONS 

FEMALES GROUP 1 0 MG/M3 

ANIMAL# OBSERVATIONS 

DAY OF - 2 

STUDY 4 7 

1591 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

1592 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

1593 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

1594 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

1595 WITHIN NORMAL LIMITS P 

TERMINAL SACRIFICE P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 7 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL# OBSERVATIONS 

DAY OF - 2 

STUDY 4 7 

2581 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

2582 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

2583 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

2584 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

2585 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE ............................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 8 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL CLINICAL OBSERVATIONS 

FEMALES GROUP 3 10000 MG/M3 

3581 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

3582 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

3583 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

3584 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

3585 WITHIN NORMAL LIMITS P 

TERMINAL SACRIFICE P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 9 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL CLINICAL OBSERVATIONS 

FEMALES GROUP 4 20000 MG/M3 

4591 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

4592 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

4593 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

4594 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

4595 WITHIN NORMAL LIMITS P 

TERMINAL SACRIFICE P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 10 

TABLE F 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL CLINICAL OBSERVATIONS 

FEMALES GROUP 6 MICRO+CONTROL 

6561 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

6562 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

6563 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

6564 WITHIN NORMAL LIMITS 

TERMINAL SACRIFICE 

6565 WITHIN NORMAL LIMITS P 

TERMINAL SACRIFICE P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 1 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 2 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

MALES GROUP 2 2000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 3 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

MALES GROUP 3 10000 MG/M3 ................................................................................................................................... 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 4 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

MALES GROUP 4 20000 MG/M3 ................................................................................................................................... 

WEEK OF STUDY 

ANIMAL# 0 1 2 3 4 

................................................................................................................................... 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 5 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

MALES GROUP 6 MICRO+CONTROL 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 6 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 7 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 8 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

FEMALES GROUP 3 10000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 9 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 10 

TABLE G 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 1 

TABLE H 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

MALES GROUP 1 0 MG/M3 

WEEK OF STUDY 

ANIMAL# 0-1 0-2 0-3 0-4 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 2 

TABLE H 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

MALES GROUP 2 2000 MG/M3 

MEAN 62 116 175 225 

S.D. 1.8 5.6 13.9 22.3 

N 5 5 5 5 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 3 

TABLE H 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

MALES GROUP 3 10000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 4 

TABLE H 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

MALES GROUP 4 20000 MG/M~ ................................................................................................................................... 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 5 

TABLE H 

GASOLINE DIPE VAPOR C0M)ENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 6 

TABLE H 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 7 

TABLE H 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

FEMALES GROUP 2 2000 MG/M3 ................................................................................................................................... 

WEEK OF STUDY 

ANIMAL# 0-1 0-2 0-3 0-4 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 8 

TABLE H 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 9 

TABLE H 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

FEMALES GROUP 4 20000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 10 

TABLE H 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 1 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 2 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 3 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

MALES GROUP 3 10000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 4 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

MALES GROUP 4 20000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 5 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AM) 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

MALES GROUP 6 MICRO+CONTROL 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 6 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

FEMALES GROUP 1 0 MG/M3 

WEEK OF STUDY 

ANIMAL# 1 2 3 4 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 7 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

FEMALES GROUP 2 2000 MG/M3 

WEEK OF STUDY 

ANIMAL# 1 2 3 4 

MEAN 

S.D. 

N 

SF-Spilled Feeder 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 8 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

FEMALES GROUP 3 10000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 9 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

FEMALES GROUP 4 20000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 10 

TABLE I 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

FEMALES GROUP 6 MICRO+CONTROL 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE I 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

MALES GROUP 1 0 MG/M3 

TYPE OF 

DEATH 

DATE OF WEEK OF STUDY 

DEATH STUDY DAY 

TERMINAL SACRIFICE 

TERMINAL SACRIFICE 

TERMINAL SACRIFICE 

TERMINAL SACRIFICE 

TERMINAL SACRIFICE 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 2 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

MALES GROUP 2 2000 MG/M3 ................................................................................................................................... 

TYPE OF DATE OF WEEK OF STUDY 

ANIMAL# DEATH DEATH STUDY DAY ................................................................................................................................... 

2071 TERMINAL SACRIFICE 11-APR-02 3 27 

2072 TERMINAL SACRIFICE 11-APR-02 3 27 

2073 TERMINAL SACRIFICE 11-APR-02 3 2 7 

2074 TERMINAL SACRIFICE 11-APR-02 3 27 

2075 TERMINAL SACRIFICE 11-APR-02 3 27 --------------------------------------------------------------------------------
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 3 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

MALES GROUP 3 10000 MG/M3 ................................................................................................................................... 

TYPE OF DATE OF WEEK OF STUDY 

ANIMAL# DEATH DEATH STUDY DAY 

................................................................................................................................... 

3071 TERMINAL SACRIFICE 11-APR-02 3 27 

3072 TERMINAL SACRIFICE 11-APR-02 3 27 

3073 TERMINAL SACRIFICE 11-APR-02 3 27 

3074 TERMINAL SACRIFICE 11-APR-02 3 27 

3075 TERMINAL SACRIFICE 11-APR-02 3 27 ................................................................................................................................... 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 4 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

MALES GROUP 4 20000 MG/M3 ................................................................................................................................... 

TYPE OF DATE OF WEEK OF STUDY 

ANIMAL# DEATH DEATH STUDY DAY 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 5 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

MALES GROUP6 MICRO+CONTROL ................................................................................................................................... 

TYPE OF DATE OF WEEK OF STUDY 

ANIMAL# DEATH DEATH STUDY DAY ................................................................................................................................... 

6051 TERMINAL SACRIFICE 11-APR-02 3 27 

6052 TERMINAL SACRIFICE 11-APR-02 3 27 

6053 TERMINAL SACRIFICE 11-APR-02 3 27 

6054 TERMINAL SACRIFICE 11-APR-02 3 27 

6055 TERMINAL SACRIFICE 11-APR-02 3 27 ------------------------------------.--------------------------------------------
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 6 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

FEMALES GROUP 1 0 MG/M3 ................................................................................................................................... 

TYPE OF DATE OF WEEK OF STUDY 

ANIMAL# DEATH DEATH STUDY DAY 

................................................................................................................................... 

1591 TERMINAL SACRIFICE 11-APR-02 3 27 

1592 TERMINAL SACRIFICE 11-APR-02 3 27 

1593 TERMINAL SACRIFICE 11-APR-02 3 27 

1594 TERMINAL SACRIFICE 11-APR-02 3 27 

1595 TERMINAL SACRIFICE 11-APR-02 3 27 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 7 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

FEMALES GROUP 2 2000 MG/M3 ----------------------------.----------------------------------------------------

TYPE OF DATE OF WEEK OF STUDY 

AN I MAL# DEATH DEATH STUDY DAY --------------------------------------------------------------------------------

2581 TERMINAL SACRIFICE 11-APR-02 3 27 

2582 TERMINAL SACRIFICE 11-APR-02 3 27 

2583 TERMINAL SACRIFICE 11-APR-02 3 2 7 

2584 TERMINAL SACRIFICE 11-APR-02 3 2 7 

2585 TERMINAL SACRIFICE 11-APR-02 3 27 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 8 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

FEMALES GROUP 3 10000 MG/M~ 

TYPE OF 

DEATH 

DATE OF WEEK OF STUDY 

DEATH STUDY DAY 

3581 TERMINAL SACRIFICE 

3582 TERMINAL SACRIFICE 

3583 TERMINAL SACRIFICE 

3584 TERMINAL SACRIFICE 

3585 TERMINAL SACRIFICE 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 9 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 
Huntingdon Life Sciences 00-6130G 

211-DIPE-S 

PAGE 10 

TABLE J 

GASOLINE DIPE VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY 

INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS 

AND 4-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

FEMALES GROUP 6 MICRO+CONTROL 

................................................................................................................................... 

TYPE OF DATE OF WEEK OF STUDY 

ANIMAL# DEATH DEATH STUDY DAY