Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

Study Title 

SATELLITE PROCEDURE 

GASOLINE TBA 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 

P23 7PX 

ENGLAND. 

HUNTINGDON LIFE SCIENCES 

LTD (PRC) STUDY NO.: 00-613 1 

HUNTINGDON LIFE SCIENCES 

LTD (ERC) STUDY NO.: APT1010 

SUBCONTRACTOR~S SPONSOR: Huntingdon Life Sciences 

Princeton Research Center (PRC) 

Mettlers Road 

East Millstone, NJ 08875-2360 

USA 

Page 1 of ?? 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

CONTENTS 

COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS ......................... 

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

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

RESPONSIBLE PERSONNEL AND SClENTlFIC APPROVAL ............................................ 

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

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

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

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

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

RESULTS ................................................................................................................................... 

CONCLUSION ........................................................................................................................... 

REFERENCES ........................................................................................................................... 

Page 

4 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

CONTENTS . continued 

TABLES 

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

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

APPENDICES 

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

2 . Animal exposure and observations data ........................................................................... 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

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), 

ENV/MClCHEM(98) 17. 

EC Commission Directive 199911 1/EC 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. 

:4: 

Date 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

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 

Process Based Inspection 

Slide scoring 

Report Audit 

Date of Inspection Date of Reporting to 

Principal Investigator 

and Test Site 

Management 

1 November 2002 1 November 2002 

25 November 2002 25 November 2002 

Date of Reporting 

to Study Director, 

Test Facility 

Management and 

Lead OA 

28 November 2002 

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 test site Quality Assurance Department. This audit 

was conducted and reported to the Principal Investigator, test site Management, Study Director, test 

facility Management and lead Quality Assurance Department 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 0101024378 

PRC - QUALITY ASSURANCE STATEMENT 

Listed below are the dates that this 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. 

Reported to 

Study Director and 

Type of Inspection Date(s) of Inspection Management 

GLP Protocol Review 9 Apr 02 9 Apr 02 

Positive Control 21 Aug 02 

Genotoxicity Dose Prep 

& Adm 

22 Aug 02 

Genotoxicity Necropsy 22 Aug 02 22 Aug 02 

Fran Jannone, B .A., RQAP-GLP 

Quality Assurance Senior Auditor 

Date 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

RESPONSIBLE PERSONNEL AND SCIENTIFIC APPROVAL 

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

Study Director 

Department of Safety Assessment, PRC. 

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

Principal Investigator 

Department of Genetic Toxicology, ERC 

Chris Davison, B.Sc., Date 

Statistician 

Department of Statistics, ERC 
Huntingdon Life Sciences (ERC) Report No: APT 010/024378 

SUMMARY 

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

Gasoline TBA 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 mg/kg 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 20" 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 TBA Vapor Condensate compared to negative control values (P>0.01 in each case). 

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

frequency of micronucleated immature erythrocytes (Pe0.001). 

It is concluded that Gasoline TBA 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 0101024378 

INTRODUCTION 

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

Condensate to induce mutagenic effects in rats following inhalation administration 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, the wealth of background 

data on this species, and 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 rnicronuclei 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 (which 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. 
Huntingdon Life Sciences (ERC) Report No: APT 010/024378 

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 Schrnid 1973). If the proportion of immature erythrocytes is found to be 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 rnicronucleus study was performed at the Department of 

Genetic Toxicology, Huntingdon Life Sciences (ERC), Eye, Suffolk, IF23 7PX, England. 

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

11 September 2002 and 28 October 2002 respectively. 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

EXPERIMENTAL PROCEDURE 

In-life phase 

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

completed on ?. 

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 mVkg bodyweight. 

Cyclophosphamide (CP, CAS # 6055-19-2, lot number 91K1176, received 20 August 2002, expiration 

30 September 2004, white powder, storage 2-8"C, purity 99.5%), 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 mg/mL 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 307 1 - 3075 3581 - 3585 

4 Test Substance 20000 408 1 - 4085 4591 - 4595 

6 Cyclophospharnide 40 (mgkg) 605 1 - 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 inhalation/exsanguination. Five males 

and five females from the positive control group were sacrificed 24 hours after CP dosing by C02 

inhalation/exsanguination. 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 
Huntingdon Life Sciences (ERC) Report No: APT 010/024378 

minutes in methanol, and aged overnight or longer prior to staining. Slides were labelled with 

experiment and animal number using a lead pencil. 

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 Romanowsky 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: 

Hydrolysed in Bouin's fluid at room temperature for approximately 28 hours. 

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

Stained in Schiff s reagent for one hour at room temperature. 

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

Counter-stained for ten minutes in very dilute (approximately 0.06 gA) aqueous Eosin 

yellowish. 

Washed for five minutes in purified water. 

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

acridine orange solution in purified water (1 mgtrnl). 

Rinsed in purified water. 

Rinsed in running tap water. 

10. Washed for 5 minutes in purified water. 

11. Air-dried. 

12. Slides were mounted with coverslips using DPX mountant. 

13. The mountant was allowed to harden at approximately 37°C. 

NB All stains and Bouin's fluid were filtered immediately prior to use to remove particulate material. 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

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 

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 Schrnid (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 1/EC of 8 March 1999 (Official Journal No. L 7718). 

Slides were hydrolysed in Bouin's fluid at room temperature for approximately 28 hours. 

The statistical analysis was performed at Huntingdon Life Sciences Ltd., Eye, Suffolk, IP23 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 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

data can be approximately fitted to a normal distribution, the results of non-parametric 

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. 

Unless there is a substantial difference in response between sexes (which occurs only rarely) results for 

the two sexes are combined to facilitate interpretation and maximise the power of statistical analysis. 

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

permutation (StatXact, CYTEL Software 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 inter-group 

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. 

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

incidence of micronucleated immature erythrocytes for the 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 (Pe0.01). 

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. 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

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 0101024378 

RESULTS 

MICRONUCLEUS TEST 

Statistical analysis was performed for both sexes and no substantial difference in response to treatment 

was recorded. The data from both sexes was pooled (Table 1) and the results are as follows: 

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, significant increases in the frequency of micronucleated immature 

erythrocytes [Pc0.001]. 

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>O.Ol]. 

Cyclophosphamide failed to cause a statistically significant decrease in the proportion [P>0.01]. 

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

It is concluded that Gasoline TBA 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 0101024378 

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-free k-sample test against ordered alternatives. Biometrics, 

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, 583-621. 

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

Statistical Association, 47, 583-621. 

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 (1996) SAS/STAT Software: Changes and Enhancements through Release 6.11. SAS 

Institute, Cary, NC, USA. 

SAS INSTlTUTE (1996) SAS/STAT Software: Changes and Enhancements for Release 6.12. SAS Institute, 

Cary, NC, USA. 
Huntingdon Life Sciences (ERC) Report No: APT 0 101024378 

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 010/024378 

TABLE 1 

Summary of results and statistical analysis 

Sampling Treatment Exposure level Proportion of ie Incidence mie Incidence mme a 

time (mg/m3' mean+SDt meankSD meankSD 

24 Hours Negative control 43k4.5 1.6k1.5 O.OkO.0 

TS 2000 46k2.9 0.9k0.6 O.OkO.0 

TS 10000 44k3.1 1.6k1.6 O.OrO.0 

TS 20000 46k4.9 1.8k1.1 0.3k0.3 

Cyclophosphamide 40 mgkg 41k4.9 13.7k5.9*** O.OkO.0 

TS Gasoline TBA Vapor Condensate 

ie Immature erythrocytes 

mie Number of micronucleated cells observed per 2000 immature erythrocytes examined 

me Mature erythrocytes 

mme Number of micronucleated cells calculated per 2000 mature erythrocytes 

SD Standard deviation 

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

permutation (one-sided probabilities): 

*** P < 0.001 (significant) 

otherwise P > 0.01 (not significant) 

t 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 group incidence mme scored x 2000 

Sum of group me scored 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

TABLE 2 

Results for individual animals - 24 hour sampling time 

Treatment Exposure level Animal ie me Proportion of Incidence Incidence 

(mg/m3) number ie rnie me 

qenative control M 1081 454 553 45 1 0 

M 1082 548 700 44 5 0 

M 1083 397 655 3 8 0 0 

M1084 518 701 42 3 0 

M 1085 501 517 49 2 0 

F1591 442 719 3 8 2 0 

F1592 511 534 49 1 0 

F 1593 551 648 46 1 0 

F 1594 510 589 46 1 0 

F 4595 579 672 46 2 0 

T S Gasoline TBA Vapor Condensate 

e Immature erythrocytes 

rnie 

me 

mme 

Number of micronucleated cells observed per 2000 immature erythrocytes 

Total number of mature erythrocytes examined for micronuclei 

Number of micronucleated mature erythrocytes observed 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

TABLE 2 - continued 

Results for individual animals - 24 hour sampling time 

Treatment Dosage Animal ie me Proportion of Incidence Incidence 

number ie rnie mrne 

Cyclophosphamide 40 mg/kg M 6051 366 70 1 34 18 0 

M 6052 390 697 36 22 0 

M 6053 494 708 41 17 0 

M 6054 453 693 40 12 0 

M 6055 346 663 34 21 0 

F 6561 447 588 43 7 0 

F 6562 436 65 1 40 11 0 

F 6563 525 594 47 14 0 

F 6564 587 773 43 4 0 

F 6565 592 643 48 11 0 

ie 

mie 

me 

rnrne 

Immature erythrocytes 

Number of micronucleated cells observed per 2000 immature erythrocytes 

Total number of mature erythrocytes examined for micronuclei 

Number of micronucleated mature erythrocytes observed 
Huntingdon Life Sciences (ERC) Report No: APT 0101024378 

APPENDIX 1 

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

Frequency of micronucleated immature erythrocytes (individual animals) 

Number of micronucleated immature erythrocytes 

(M IE per 2000) 

Individual man 0.79 

Historical negative control values (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 0101024378 

APPENDIX 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 

(MIE 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) 

Cyclophospham ide 

Number of micronucleated immature erythrocytes (MIEper 2000) 

Group man 39.9 
Huntingdon Life Sciences (ERC) Report No: APT 010/024378 

APPENDIX 2 

ANIMAL EXPOSURE AND OBSERVATIONS DATA 
Huntingdon Life Sciences 00-6 13 1 Page 1 

21 1-TBA-S Draft Final Re~ort 

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. 

Animal Exposure and Animal Data 

Preface 

STUDY DATES: Date of Animal Receipt: 15 July 2002 

Experimental Initiation Date: 26 July 2002 (in-life) 

Experimental Completion Date: 22 August 2002 (in-life) 

Draft Report Date: 2 June 2004 

Appendix 2 

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 (pretest only) ............................................................ C. Mean Body Weights (grams) ........................................................................................... D. Mean Body Weight Change from Baseline (grams) ........................................................ E. Mean Feed Consumption Values (gramskglday) ............................................................ 

F. Individual Clinical Observations (pretest only) ............................................................... G. Individual Body Weights (grams) .................................................................................... H. Individual Body Weight Change from Baseline (grams) ................................................. 

I. Individual Feed Consumption Values (grams/kg/day) ..................................................... J. Animal Termination History ............................................................................................ 
Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-61 31 

Chamber Monitoring Results 

Cumulative Exposure Record 
Table A GASOLINE TEA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-61 31 

Chamber Monitoring Results 

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. 

Cumulative Exposure Record 

Group IB - 0 (air only) mglm3 

Nominal 

(ms/m3) 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

Mean 

(ms/m3) 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

alytical Chamber Concentration 

Individual 

I 

MMAD 

Olm) 

3.102 

2.850 

0.7383 

3.732 

Particle Size 

terminal 

GSD 

ns 

TMC 

(mslm3) 

7.27E-03 

5.51 E-03 

2.12E-02 

6.47E-03 

Chamber Environment 

Mean 

Temperature 

("C) 

23 

24 

24 

25 

25 

25 

25 

25 

24 

24 

24 

24 

24 

25 

25 

25 

25 

25 

24 

Humidity 

("w 

55 

55 

53 

53 

56 

56 

56 

53 

45 

49 

50 

52 

53 

54 

54 

56 

54 

52 

56 

56 

53.4 

2.8 
Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131 

Chamber Monitoring Results 

Date Exposure 

Number 

Mean 

Nominal 

(mglm3) 

1830 

2090 

2070 

1990 

201 0 

2150 

2110 

2080 

1950 

2020 

21 60 

2020 

2020 

21 90 

2090 

2090 

21 40 

2200 

2070 

21 30 

Cumulative Exposure Record 

Grou~ HA - 2.000 ma/m3 

Mean 

(mglm") 

1840 

2000 

1935 

1933 

1953 

1975 

2005 

1970 

1963 

21 35 

21 88 

1953 

1845 

2043 

2023 

201 3 

2070 

2090 

2068 

21 83 

alytical Chamber Concentration 

Individual MMAD 

Olm) 

2.31 2 

4.144 

0.7525 

0.8197 

Particle Size 

ztermina 

GSD 

Ins 

TMC 

(mglm3) 

4.69E-03 

9.38E-03 

2.35E-02 

2.61 E-03 

Chamber En 

M ea 

TemDerature 

ronment 

Humidity 

("w 

50 

50 

48 

49 

51 

51 

50 

48 

40 

44 

44 

46 

47 

47 

48 

50 

49 

48 

48 

52 

48.0 

2.8 
Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131 

Chamber Monitoring Results 

Cumulative Exposure Record 

Group IIB - 2,000 mslm3 

Chamber El 

Me; 

ronment 

Humidity 

("w 

52 

53 

50 

52 

54 

55 

55 

5 1 

43 

46 

47 

50 

49 

50 

52 

53 

53 

50 

54 

56 

Particle Size 

Nominal alytical Chamber Concentration Determinations 

(mglm3) 

1830 

2090 

2070 

1990 

201 0 

2150 

21 10 

2080 

1950 

2020 

21 60 

2020 

2020 

21 90 

2090 

2090 

21 40 

2200 

2070 

21 30 

2071 

88 

Date Exposure Temperature 

Number Mean 

(mslm3) 

1928 

2105 

2060 

1935 

1830 

2048 

21 23 

2020 

2018 

1970 

2203 

1968 

2088 

21 65 

2008 

1995 

2028 

2073 

2030 

2025 

Individual MMAD 

Olm) 

3.582 

2.398 

0.7703 

0.8346 

GSD TMC 

~~~~m3) 

6.02E-03 

2.09E-03 

1.73E-02 

2.64E-03 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 
Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131 

Date Exposure 

Number 

21-Aug-02 ( 20 

Mean 

S.D. 

Chamber Monitoring Results 

Cumulative Exposure Record 

Group lllA - 10,000 mg/m3 

Nominal 

(mg/m3) 

101 00 

9960 

10200 

10400 

10300 

10400 

9790 

10000 

9580 

10500 

991 0 

9770 

9770 

10000 

9560 

9660 

9850 

10100 

10600 

10000 

10020 

302 

Analytical Chamber Concentration 

Mean I Individual 

Particle Size 

Determinati 

MMAD I GSD 

IS 

TMC 

(mg/m3) 

7.41 E-03 

3.72E-03 

2.45E-02 

2.23E-03 

Chamber Environment 

Mean 

Temperature 

("C) 

23 

24 

24 

23 

23 

23 

23 

23 

24 

24 

24 

24 

24 

23 

23 

23 

23 

23 

24 

24 

23.5 

0.5 

Humidity 

("w 

50 

50 

50 

50 

52 

52 

52 

49 

40 

44 

44 

47 

47 

49 

51 

52 

52 

48 

50 

51 

49.0 

3.2 
Table A GASOLINE TEA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131 

Chamber Monitoring Results 

Exposure 

Number 

Day 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 

S.D. 

Date 

Cumulative Exposure Record 

Group lllB - 10,000 mg/m3 

I 

Nominal 

(mslm3) 

101 00 

9960 

10200 

10400 

10300 

10400 

9790 

10000 

9580 

10500 

991 0 

9770 

9770 

10000 

9560 

9660 

9850 

10100 

10600 

10000 

10020 

302 

Mean 

(ms/m3) 

1001 0 

10500 

10020 

10000 

10090 

10160 

10100 

10240 

lo620 

10750 

11150 

lo260 

10680 

lo280 

10240 

9605 

10330 

9880 

lo380 

9955 

Analvtical Chamber Concentration 

Individual I MMAD I GSD I TMC 

Particle Size 

Determinations 

Chamber Environment 

Temperature 

("C) 

23 

23 

23 

24 

24 

24 

24 

24 

24 

24 

24 

23 

24 

24 

24 

24 

24 

24 

24 

23 

23.8 

0.4 

1 

Humidity 

(W 

47 

48 

49 

47 

48 

48 

48 

46 

39 

42 

41 

44 

45 

45 

46 

47 

48 

45 

47 

48 

45.9 

2.7 
Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131 

Chamber Monitoring Results 

Cumulative Exposure Record 

Grow IVA - 20.000 malm3 

Date Exposure 

Number 

Mean 

~alytical Chamber Concentration 

Individual 

Nominal 

(ms/m3) 

18900 

19100 

18400 

18500 

18900 

19100 

18900 

18800 

19100 

19700 

18800 

18700 

18800 

19000 

19500 

19400 

19100 

19300 

19200 

18800 

19000 

321 

I 

MMAD 

Olm) 

1.689 

2.505 

0.7685 

1.301 

1 

Mean 

(mglm3) 

20500 

20650 

19800 

20380 

19500 

19480 

19250 

19530 

19950 

21380 

20530 

19480 

19200 

18500 

19000 

19430 

20630 

19500 

20250 

20300 

I Chamber Environment 

Particle Size I Mean 

terminations ( Temperature I Humidity 

GSD I TMC I I 
Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131 

Chamber Monitoring Results 

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. 

Cumulative Exposure Record 

Group IVB - 20,000 mglm3 

Nominal 

(ms/m3) 

18900 

191 00 

18400 

18500 

18900 

19100 

18900 

18800 

191 00 

19700 

18800 

18700 

18800 

19000 

19500 

19400 

191 00 

19300 

19200 

18800 

19000 

321 

Mean 

(mg/m3) 

19550 

19650 

20080 

19400 

20250 

201 80 

19600 

19880 

19880 

20650 

19330 

20300 

20050 

19750 

19980 

20580 

20650 

20330 

20030 

20500 

alytical Chamber Concentration 

Individual 

I 

MMAD 

Olm) 

1.032 

1.793 

0.7982 

0.9155 

Particle Size 

1s 

TMC 

(mglm3) 

1 .WE-03 

2.30E-03 

2.51 E-02 

3.08E-03 

Chamber En 

Mea 

Temperature 

("C) 

24 

24 

24 

26 

26 

25 

26 

26 

25 

25 

24 

24 

25 

26 

25 

25 

25 

26 

25 

24 

ronment 

Humidity 

("w 

52 

54 

52 

5 1 

51 

53 

52 

51 

43 

47 

46 

48 

50 

50 

51 

55 

53 

50 

52 

55 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

TABLE B 

GASOLINE TBA 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 

PAGE 1 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE B 

GASOLINE TBA 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 ................................................................................................................................... 

DAY OF STUDY 

GROUP# -4 TOTAL ................................................................................................................................... 

# OF ANIMALS EXAMINED 15 

2 5 

3 5 

4 5 

NORMAL 

WITHIN NORMAL LIMITS 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE C 

GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION 

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

DAY 

DAY 

DAY 

DAY 

DAY 

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 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE C 

GASOLINE TBA 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 BODY WEIGHTS (GRAMS) ................................................................................................................................... 

DOSE GROUP: I I I I11 I V VI 

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

DAY -3 MEAN 

S.D. 

N 

DAY 4 

DAY 11 

DAY 18 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

DAY 25 MEAN 237 222 235 235 238 

S.D. 22.9 22.0 15.0 10.2 10.7 

N 5 5 5 5 5 ................................................................................................................................... 

No statistically significant differences 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE D 

GASOLINE TBA 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 BODY WEIGHT CHANGE FROM BASELINE (GRAMS) ................................................................................................................................... 

DOSE GROUP: I I I I I I IV VI 

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

DAY -3 TO 11 MEAN 

S.D. 

N 

DAY -3T0 18 MEAN 

S.D. 

N 

DAY -3T0 25 

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

MEAN 

S.D. 

N 

No statistically significant differences 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE D 

GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION 

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

DAY -3T0 11 

DAY -3T0 18 

MEAN 

S.D. 

N 

MEAN 

S.D. 

N 

DAY -3T0 25 MEAN 107 

S.D. 16.8 

N 5 

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

No statistically significant differences 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE E 

GASOLINE TBA 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: I 

DOSE LEVEL (MG/M3) : 0 ....................................................... 

DAY 4 MEAN 

S.D. 

N 

DAY 11 MEAN 

S.D. 

N 

DAY 18 MEAN 

S.D. 

N 

DAY 25 

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

MEAN 

S.D. 

N 

No statistically significant differences 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE E 

GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION 

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

DAY 4 MEAN 

S.D. 

N 

DAY 11 MEAN 

S.D. 

N 

DAY 18 MEAN 

S.D. 

N 

DAY 25 MEAN 

S.D. 

N 

Statistical key: * = pc0.05 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

APPENDIX F 

GASOLINE TBA 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 WEEKLY CLINICAL OBSERVATIONS 

MALES GROUP I 0 MG/M3 ................................................................................................... 

1081 WITHIN NORMAL LIMITS P 

1082 WITHIN NORMAL LIMITS P 

1083 WITHIN NORMAL LIMITS P 

1084 WITHIN NORMAL LIMITS P 

1085 WITHIN NORMAL LIMITS P ................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

APPENDIX F 

GASOLINE TBA 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 GROUP I1 2000 MG/M3 .......................................... 

ANIMAL# OBSERVATIONS 

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

2071 WITHIN NORMAL LIMITS 

2072 WITHIN NORMAL LIMITS 

2073 WITHIN NORMAL LIMITS 

2074 WITHIN NORMAL LIMITS 

2075 WITHIN NORMAL LIMITS 

INDIVIDUAL WEEKLY CLINICAL OBSERVATIONS 

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

DAY OF 

STUDY -. 

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

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 3 

APPENDIX F 

GASOLINE TBA 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 WEEKLY CLINICAL OBSERVATIONS 

MALES GROUP I11 10000 MG/M3 ................................................................................................................................... 

3071 WITHIN NORMAL LIMITS P 

3072 WITHIN NORMAL LIMITS P 

3073 WITHIN NORMAL LIMITS P 

3074 WITHIN NORMAL LIMITS P 

3075 WITHIN NORMAL LIMITS P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 4 

APPENDIX F 

GASOLINE TBA 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 WEEKLY CLINICAL OBSERVATIONS 

MALES GROUP IV 20000 MG/M3 .................................................................................................................... 

4081 WITHIN NORMAL LIMITS P 

4082 WITHIN NORMAL LIMITS P 

4083 WITHIN NORMAL LIMITS P 

4084 WITHIN NORMAL LIMITS P 

- - 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 5 

APPENDIX F 

GASOLINE TBA 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 WEEKLY CLINICAL OBSERVATIONS 

MALES GROUP VI MICRO+CONTROL 

ANIMAL# OBSERVATIONS 

DAY OF 

STUDY 4 

6051 WITHIN NORMAL LIMITS P 

6052 WITHIN NORMAL LIMITS P 

6053 WITHIN NORMAL LIMITS P 

6054 WITHIN NORMAL LIMITS P 

6055 WITHIN NORMAL LIMITS P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 6 

APPENDIX F 

GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION 

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

1591 WITHIN NORMAL LIMITS 

1592 WITHIN NORMAL LIMITS 

1593 WITHIN NORMAL LIMITS 

1594 WITHIN NORMAL LIMITS 

1595 WITHIN NORMAL LIMITS 

INDIVIDUAL WEEKLY CLINICAL OBSERVATIONS 

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

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

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 7 

APPENDIX F 

GASOLINE TBA 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 WEEKLY CLINICAL OBSERVATIONS 

FEMALES GROUP I1 2000 MG/M3 ................................................................................................................................... 

2581 WITHIN NORMAL LIMITS P 

2582 WITHIN NORMAL LIMITS P 

2583 WITHIN NORMAL LIMITS P 

2584 WITHIN NORMAL LIMITS P 

2585 WITHIN NORMAL LIMITS P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 8 

APPENDIX F 

GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION 

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WFEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL WEEKLY CLINICAL OBSERVATIONS 

FEMALES GROUP I11 10000 MG/M3 ................................................................................................................................... 

3581 WITHIN NORMAL LIMITS P 

3582 WITHIN NORMAL LIMITS P 

3583 WITHIN NORMAL LIMITS P 

3584 WITHIN NORMAL LIMITS P 

3585 WITHIN NORMAL LIMITS P ............................................................................................ 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 9 

APPENDIX F 

GASOLINE TBA 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 WEEKLY CLINICAL OBSERVATIONS 

FEMALES GROUP IV 20000 MG/M3 .................................................................................................. 

DAY OF 

ANIMAL# OBSERVATIONS STUDY ............................................................ 

4591 WITHIN NORMAL LIMITS 

4592 WITHIN NORMAL LIMITS 

4593 WITHIN NORMAL LIMITS 

4594 WITHIN NORMAL LIMITS 

4595 WITHIN NORMAL LIMITS ............................................................ 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 10 

APPENDIX F 

GASOLINE TBA 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 WEEKLY CLINICAL OBSERVATIONS 

FEMALES GROUP VI MICRO+CONTROL .............................................................................................. 

DAY OF 

ANIMAL# OBSERVATIONS STUDY 4 .............................................................................................. 

6561 WITHIN NORMAL LIMITS P 

6562 WITHIN NORMAL LIMITS P 

6563 WITHIN NORMAL LIMITS P 

6564 WITHIN NORMAL LIMITS P 

6565 WITHIN NORMAL LIMITS P ................................................................................................................................... 

CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE G 

GASOLINE TBA 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 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE G 

GASOLINE TBA 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 I1 2000 MG/M~ 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 3 

TABLE G 

GASOLINE TBA 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 I11 10000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 4 

TABLE G 

GASOLINE TBA 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 IV 20000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 5 

TABLE G 

GASOLINE TBA 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 VI MICRO+CONTROL 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 6 

TABLE G 

GASOLINE TBA 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 I 0 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 7 

TABLE G 

GASOLINE TBA 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 I1 2000 MG/M3 

DAY OF STUDY 

ANIMAL# -3 4 11 18 25 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 8 

TABLE G 

GASOLINE TBA 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 I11 10000 MG/M3 

MEAN 

S.D. 

N 

OF STUDY 

4 11 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 9 

TABLE G 

GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION 

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMIWNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 10 

TABLE G 

GASOLINE TEA 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 VI MICRO+CONTROL 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE H 

GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION 

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMIWNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

MALES GROUP I 0 MG/M~ 

DAY OF STUDY 

ANIMAL# -3-4 -3-11 -3-18 -3-25 

1081 56 107 161 198 

1082 55 111 167 215 

1083 55 108 165 207 

1084 63 123 182 231 

1085 4 9 98 143 182 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE H 

GASOLINE TBA 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 I1 2000 MG/M3 

DAY OF STUDY 

ANIMAL# -3-4 -3-11 -3-18 -3-25 

2071 55 108 159 199 

2072 56 112 165 213 

2073 56 115 166 213 

2074 61 116 166 200 

2075 63 109 155 187 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 3 

TABLE H 

GASOLINE TBA 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 00-6131G 

Genotoxicity Sub-Group 

PAGE 4 

TABLE H 

GASOLINE TBA 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 IV 20000 MG/M3 ................................................................................................................................... 

DAY OF STUDY 

ANIMAL# -3-4 -3-11 -3-18 -3-25 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 5 

TABLE H 

GASOLINE TBA 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 VI MICRO+CONTROL ................................................................................................................................... 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 6 

TABLE H 

GASOLINE TBA 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 00-6131G 

Genotoxicity Sub-Group 

PAGE 7 

TABLE H 

GASOLINE TBA 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 00-6131G 

Genotoxicity Sub-Group 

PAGE 8 

TABLE H 

GASOLINE TBA 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 I11 10000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 9 

TABLE H 

GASOLINE TBA 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 00-6131G 

Genotoxicity Sub-Group 

PAGE 10 

TABLE H 

GASOLINE TBA 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 VI MICRO+CONTROL 

DAY OF STUDY 

ANIMAL# -3-4 -3-11 -3-18 -3-25 

6561 33 58 81 9 4 

6562 3 8 58 89 112 

6563 43 78 107 126 

6564 3 9 66 85 108 

6565 32 59 88 109 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE I 

GASOLINE TBA 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 I 0 MG/M3 

DAY OF STUDY 

4 11 18 25 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE I 

GASOLINE TBA 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 I1 2000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 3 

TABLE I 

GASOLINE TBA 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 I11 10000 MG/M3 

DAY 

ANIMAL# 4 

MEAN 

S.D. 

N 

OF STUDY 

11 18 25 

SF=Spilled Feeder 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 4 

TABLE I 

GASOLINE TBA 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 IV 20000 MG/M3 

DAY OF STUDY 

4 11 18 2 5 

MEAN 128 107 93 83 

S.D. 8.1 4.4 1.5 1.8 

N 5 5 5 5 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 5 

TABLE I 

GASOLINE TBA 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 VI MICRO+CONTROL 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 6 

TABLE I 

GASOLINE TBA 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 I 0 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 7 

TABLE I 

GASOLINE TBA 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 00-6131G 

Genotoxicity Sub-Group 

PAGE 8 

TABLE I 

GASOLINE TBA 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 I11 10000 MG/M3 ................................................................................................................................... 

OF STUDY 

11 18 2 5 ................................................................................. 

105 97 91 

106 98 9 7 

105 110 8 8 

145 SF 9 2 

108 103 9 3 

MEAN 

S.D. 

N 

SF=Spilled Feeder 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 9 

TABLE I 

GASOLINE TBA 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 IV 20000 MG/M3 ................................................................................................................................... 

DAY OF STUDY 

ANIMAL# 4 11 18 25 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 10 

TABLE I 

GASOLINE TBA 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 VI MICRO+CONTROL 

DAY OF STUDY 

ANIMAL# 4 11 18 25 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

GASOLINE 

TOXICITY 

MALES GROUP I 0 MG/M3 

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

TYPE OF 

ANIMAL# DEATH 

PAGE 1 

TABLE J 

TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION 

STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE J 

GASOLINE TBA 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 I1 2000 MG/M3 

TYPE OF 

DEATH 

DATE OF WEEK OF STUDY 

DEATH STUDY DAY 

2071 TERMINAL SACRIFICE 

2072 TERMINAL SACRIFICE 

2073 TERMINAL SACRIFICE 

2074 TERMINAL SACRIFICE 

2075 TERMINAL SACRIFICE 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group ' 

PAGE 3 

TABLE J 

GASOLINE TBA 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 I11 10000 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 00-6131G 

Genotoxicity Sub-Group 

PAGE 4 

TABLE J 

GASOLINE TBA 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 

Genotoxicity Sub-Group 

GASOLINE 

TOXICITY 

IN 

MALES GROUP VI MICRO+CONTROL 

PAGE 5 

TABLE J 

TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION 

STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

ANIMAL TERMINATION HISTORY 

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

TYPE OF DATE OF WEEK OF STUDY 

ANIMAL# DEATH DEATH STUDY DAY 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 6 

TABLE J 

GASOLINE TBA 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 I 0 MG/M3 

1591 TERMINAL SACRIFICE 22-AUG-02 3 27 

1592 TERMINAL SACRIFICE 22-AUG-02 3 27 

1593 TERMINAL SACRIFICE 22-AUG-02 3 27 

1594 TERMINAL SACRIFICE 22-AUG-02 3 27 

1595 TERMINAL SACRIFICE 22-AUG-02 3 27 ................................................................................................................................... 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 7 

TABLE J 

GASOLINE TBA 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 I1 2000 MG/M3 ................................................................................................................................... 

TERMINAL SACRIFICE 22-AUG-02 

TERMINAL SACRIFICE 22-AUG-02 

TERMINAL SACRIFICE 22-AUG-02 

TERMINAL SACRIFICE 22-AUG-02 

TERMINAL SACRIFICE 22-AUG-02 .................................................................. 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 8 

TABLE J 

GASOLINE TBA 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 I11 10000 MG/M~ 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 9 

TABLE J 

GASOLINE TBA 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 IV 20000 MG/M3 ................................................................................................................................... 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 10 

TABLE J 

GASOLINE TBA 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 VI MICRO+CONTROL ................................................................................................................................... 

TYPE OF DATE OF WEEK OF STUDY 

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

6561 TERMINAL SACRIFICE 22-AUG-02 3 27 

6562 TERMINAL SACRIFICE 22-AUG-02 3 27 

6563 TERMINAL SACRIFICE 22-AUG-02 3 27 

6564 TERMINAL SACRIFICE 22-AUG-02 3 27 

6565 TERMINAL SACRIFICE 22-AUG-02 3 27