## HuntingdonLife Sciences(ERC) ReportNo: 

## APT 0101024378

Study 

## Title SATELLITE PROCEDURE GASOLINE TBAVAPOR CONDENSATE 

## RATMICRONUCLEUS TEST TEST GUIDELINES: 

## US EPA Micronucleus Assay 79.64, 

## CFR Vol. 59, No. 122,27June 1994. 

## US EPA (1998) Health Effects Test Guidelines; OPPTS 870.5395 MammalianErythrocyte Micronucleus Test. 

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



## SUBCONTRACTOR: Huntingdon Life Sciences Ltd., Eye Research Centre 

## (ERC) Eye, Suffolk P237PX ENGLAND.
HUNTINGDONLIFE 

## SCIENCES LTD

## (PRC) STUDYNO.: 

## 00-6131 HUNTINGDONLIFE 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 1of ??

## HuntingdonLife Sciences (ERC) Report No: APT 0101024378 

## CONTENTS

COMPLIANCEWITH GOOD LABORATORY PRACTICE STANDARDS


## ......................... ERC

. 

## QUALITY ASSURANCE STATEMENT


## ...................................................................... PRC

. 

## QUALITY ASSURANCE STATEMENT


## ...................................................................... RESPONSIBLE PERSONNELAND SClENTlFICAPPROVAL


## ............................................

................................................................................................................................ SUMMARY INTRODUCTION 

## ...................................................................................................................... EXPERIMENTAL PROCEDURE

## ............................................................................................. ASSESSMENT OFRESULTS 

## .................................................................................................. MAINTENANCEOFRECORDS

## .............................................................................................. RESULTS


## ................................................................................................................................... CONCLUSION

# ........................................................................................................................... REFERENCES 

## ........................................................................................................................... Page 4

## HuntingdonLife Sciences(ERC) Report No: APT 0101024378



## CONTENTS 

# . continued TABLES 1

# . Summary ofresults and statisticalanalysis

## ...................................................................... 2


## . Results for individualanimals

. 

## 24 hour samplingtime


## .................................................. APPENDICES 

## 1



## . Historical negative and positive controlvalues 

## ...............................................................

........................................................................... 2

# . Animal exposureand observations data

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

## COMPLIANCEWITH GOODLABORATORY PRACTICE STANDARDS
The slide evaluation phase of the study described in this report was conducted in compliance with the followingGoodLaboratory Practice standardsand 

## I consider the data generated to be valid. The 

## UK Good Laboratory PracticeRegulations 1999(StatutoryInstrument No. 3106). OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MClCHEM(98)17. EC CommissionDirective 1999111/ECof 8 March 1999 (Official Journal No. 

## L 7718). US EPA 79.60, CFR Vol. 59,No. 122,27June 1994. No complianceis 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 SciencesLtd., 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 stabilityof the positive control articlemixture has not been determinedby the Testing Facility.

Gary M. Hoffman, 

## B.A.,

## D.A.B.T., Study Director, Huntingdon Life Sciences Ltd., PRC. :4: Date

## HuntingdonLife Sciences(ERC) ReportNo: 

## APT 0101024378 

## ERC

# - QUALITY ASSURANCE STATEMENT The following inspection and audit have been carried out in relation to the slide evaluationphase of this study: 

## Study Phase

ProcessBased Inspection Slide scoring 

## Report Audit Date of Inspection Date of Reporting to Principal Investigator and Test Site Management

1

## November 2002 1November 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 appropriateCompany Management as indicated above. 

## Report Audit:This appendixhas been audited by the test site Quality AssuranceDepartment. This audit was conducted and reported to the Principal Investigator, test site Management, Study Director, test facilityManagement and lead Quality Assurance Department as indicated above. Study based inspectionswere 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, HuntingdonLife Sciences Ltd. Date

## Huntingdon Life Sciences (ERC) Report No: 

## APT 0101024378 

## PRC

# - QUALITY ASSURANCE STATEMENT Listedbelow are the dates that this studywas inspectedby 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 Directorand Type ofInspection Date(s)ofInspection Management GLP Protocol Review 

## 9 Apr 02 

## 9 Apr 02 Positive Control 21 Aug 02 Genotoxicity Dose Prep & 

## Adm 22 Aug 02 GenotoxicityNecropsy 22 Aug 02 22 Aug 02

Fran Jannone,

## B.A.,RQAP-GLP QualityAssurance SeniorAuditor Date

## Huntingdon Life Sciences (ERC)ReportNo: 

## 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

## HuntingdonLife 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 occasionby 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 substancegroups 24 hours after the 20" exposure and fromthe 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 assessedby examinationof at least 1000 erythrocytes from each animal. A record of the incidence of micronucleated mature erythrocytes was alsokept. 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 Condensatecompared to negative control values (P>0.01in each case). The positive control compound, Cyclophosphamide, produced large, significant increases in the frequencyof micronucleated immature erythrocytes(Pe0.001). 

## It is concluded that GasolineTBA Vapor Condensatedid not show any evidence of causing chromosome damage or bone marrow cell toxicity when administered by inhalation exposure in this in 

## vivo 

## test procedure.

## HuntingdonLife 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. Theprocedures used were based on the recommendations of the following guidelines: US EPA MicronucleusAssay 79.64, CFR Vol. 59,No. 122,27June 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 accuratelymodelled in an 

## in vitro system. Young adult rats are chosen for use because of the high rate of cell divisionin 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 causedby 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). Micronucleatedimmature erythrocytesappear in thebone marrow approximately 24 hours after inductionof chromosome damage. These immature erythrocytescanbe differentiatedby a variety of stainingtechniques 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 incidenceof micronucleated immature erythrocytesis indicativeof recent exposure to a chromosome-damaging agent. A simultaneous marked increase in the incidence of micronucleated mature erythrocytes is not expected and may be indicativeofmicronucleus-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 laggingchromosomes at anaphase which may not be incorporated into the daughter nuclei. These lagging chromosomes are not excluded from the erythroblastwith the main nucleus and hence also give rise to micronuclei.

## HuntingdonLife 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 erythrocytesis 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 indicativeof a cytostaticor 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 11September 2002 and 28 October 2002 respectively.

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

## EXPERIMENTAL PROCEDURE In-lifephase 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 10mVkg bodyweight. Cyclophosphamide (CP, CAS # 

## 6055-19-2, lot number 91K1176, received 20 August 2002, expiration 30 September 2004, white powder, storage2-8"C, purity 99.5%), was obtained fromthe 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 experimentaldesign 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 4081

## - 4085 4591 - 

### 4595 6 

## Cyclophospharnide 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 exposureperiod 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 supernatantdrawn off, leaving a small amount of serumwith 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

## HuntingdonLife Sciences (ERC) ReportNo: 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, IP237PX, England for slide staining and analysis. The remaining 2 smears and the cell pellet (refrigerated)were held in reserve at PRC in case of technicalproblems 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 stainsDNA-containing bodies deep purple while leaving mast cell granules unstained. The method also allows reasonable differentiationof mature and immature erythrocytesand produces permanent preparations. Two slides from each animal were stained as follows: Hydrolysed in Bouin's fluid atroom temperature for approximately 28 hours. Washed threetimes in

## purified water 

## (5minutes per wash). Stained in Schiffs reagent for onehour atroom 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 fiveminutes in purified water. Stained for 30 minutes in Mayer's Haemalum diluted 

## 9 volumes: 1 volume with aqueous acridine orange solution

## inpurified water (1mgtrnl). Rinsed in purified water. Rinsedinrunningtap water. 10. Washed for

## 5 minutes 

## in purified water. 11. Air-dried. 12. Slideswere mounted with coverslipsusing DPX mountant. 13. Themountant was allowedto 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) ReportNo: 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 technicalproblems with the first smear. Micronuclei are identifiedby the following criteria: Large enough to discernmorphological characteristics Should possess a generally rounded shapewith a clearly defined outline Shouldbe deeply stained and similar in colourto the nuclei of other cells - 

## not black Should lie in the same focal plane as the cell Lack internal structure,ie 

## they arepyknotic 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 recommendedby 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 PracticeRegulations 1999 (StatutoryInstrument No. 3106), the OECD Principles of Good Laboratory Practice (as revised in 1997),ENV/MC/CHEM(98)17 and the EC CommissionDirective 1999111/ECof 8 March 1999(Official JournalNo. L 7718). Slideswere 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 resultsbecause: They are suited to analysis of data consisting of discretelintegervalues with ties such as the incidenceof 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 analysisof variance are very similar). 'Outliers' are frequently found in the proportionof immature erythrocytesforboth 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 interpretationand 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 straightforwardpermutation 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 comparedwith 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 controlrange. 

## 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 significantdose-related decrease in the proportionof 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 archivingmay be disposedof.

## HuntingdonLife Sciences(ERC) ReportNo: 

## APT 

## 0101024378 All other samples and specimens and all raw data will be retained by Huntingdon Life SciencesPRC 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 subjectto a reasonablefee 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.

## HuntingdonLife Sciences(ERC) Report No: APT 0101024378
RESULTS

MICRONUCLEUSTEST 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 erythrocytecounts (mie) The test substance did not cause any statistically significant increases in the number of micronucleated immature erythrocytes [P>0.011. 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]. Cyclophosphamidefailed 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 Condensatecompared to negative control values (P>0.01 in each case). It is concluded that Gasoline

## TBA Vapor Condensatedid not show any evidence of causingchromosome damage or bone marrow cell toxicity when administeredby 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 ofthe 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-freek-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 ofthe American StatisticalAssociation,47, 583-621. KRUSKAL, W.H. and WALLIS, W.A. (1953) Errata for Kruskal-Wallis (1952). Journal of theAmerican StatisticalAssociation,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 sixmammalian species, evaluatedby 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 ProtectionAgency Gene-Tox Program. Mutation Research, 239,29. MORRISON, V. and ASHBY, J. (1995) High resolutionrodent bone marrow micronucleusassays of 1,2- dimethylhydrazine :

## implicationof 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/STATSoftware: Changes and Enhancementsfor Release 6.12. SAS Institute, Cary, NC, USA.

## HuntingdonLife 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.Publishedby Plenum Press,New York.
von LEDEBUR, 

## M. and SCHMID,

## W. (1973) The rnicronucleus test. Methodological aspects. Mutation Research, 

## 19, 109. WILCOXON,F. (1945). Individualcomparisonsby ranking methods. Biometries Bulletin, 

## 1,80-83.

## HuntingdonLife Sciences(ERC) Report No: APT 010/024378



## TABLE 1 

## Summary of 

## resultsand statistical analysis Sampling Treatment Exposure level Proportionof ie Incidencemie 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 GasolineTBA Vapor Condensate ie Immature erythrocytes mie Number of micronucleated cells observedper 2000 

## immature erythrocytesexamined me Mature erythrocytes mme Number of micronucleated cells calculatedper 2000 

## mature erythrocytes SD Standarddeviation Results of statistical analysisusing the appropriatenonparametric method of analysisbased on permutation (one-sidedprobabilities): 

# *** 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 calculationof incidencemme (group mean): Sumof group incidence

## mme scored 

## x 2000 

## Sumof group 

## me scored

## HuntingdonLife Sciences (ERC) Report No: APT 0101024378


## TABLE2 

## Results for individual animals

# - 24 

## hour sampling time Treatment Exposure level Animal ie me Proportion of Incidence Incidence (mg/m3) number ie rnie me qenativecontrol 

### 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 

### 38 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 observedper 

### 2000 immature erythrocytes Total number of mature erythrocytes examined for micronuclei Number of micronucleated mature erythrocytes observed

## HuntingdonLife Sciences (ERC) Report No: APT 

## 0101024378



## TABLE2

# - continued Resultsfor 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 366 701 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 436 651 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 observedper 

### 2000 immature erythrocytes Total number of mature erythrocytesexamined for micronuclei Number of micronucleated mature erythrocytes observed

## HuntingdonLife Sciences

## (ERC)ReportNo: 

## APT 0101024378



## APPENDIX 1
Historical negative controlvalues (1 July 2000

# - 30June 2002) Frequencyof micronucleatedimmatureerythrocytes(individualanimals)

Number of micronucleated immature erythrocytes (MIE

### per 2000) Individual man0.79 

### Historicalnegative controlvalues (1 July2000

# - 30June 2002) Frequencyof micronucleatedimmatureerythrocytes(Group meanvalues)

Number of micronucleated immature erythrocytes (MIEper 2000) 

### Group man0.73

## Huntingdon Life Sciences

## (ERC) Report No: APT 

## 0101024378 

## APPENDIX 1

# -continued
Historical positive control values (1 July 2000

# - 30 June 2002) Frequencyof micronucleated immature erythrocytes (Individualanimals) 

### Cyclophosphamide

Number of micronucleated immature erythrocytes (MIE per 2000 

### cells) Individualmean 38.7



### Historical positivecontrolvalues 

### (1 July 2000

# - 30 June 2002) Frequencyof micronucleated immature erythrocytes (Group meanvalues) 

### Cyclophosphamide

Number of micronucleated immature erythrocytes (MIEper 2000) 

### Group 

### man 39.9

## HuntingdonLife Sciences(ERC) Report No: APT 

## 010/024378 

## APPENDIX 

## 2 ANIMALEXPOSURE AND OBSERVATIONS DATA

## Huntingdon Life Sciences 00-6131 Page 1 211-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 detailsof the procedures used for test atmosphere 

## generationkharacterization and animal evaluations. Animal Exposure and Animal Data Preface



# STUDY DATES: Date of Animal Receipt: 15July 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. 

# TABLEOF CONTENTS TABLES



# ........................................................................................... A. Chamber Monitoring Results B. Summaryof Clinical Observations(pretest only)

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

# ........................................................................................... D. Mean Body Weight Change from Baseline (grams) 

# ........................................................ E. Mean Feed ConsumptionValues (gramskglday)

# ............................................................ F. Individual Clinical Observations(pretest only) 

# ............................................................... G. IndividualBody Weights (grams)

# ....................................................................................

................................................. H. IndividualBody Weight Change from Baseline (grams) 

## I. Individual Feed ConsumptionValues (grams/kg/day)

# ..................................................... J. Animal TerminationHistory

# ............................................................................................Table A GASOLINETBA VAPOR CONDENSATE:A 13-WEEK WHOLE-BODY INHALATIONTOXICITY STUDY IN RATS 00-61 31 Chamber Monitoring Results Cumulative Exposure RecordTable A GASOLINETEA VAPOR CONDENSATE: A 13-WEEKWHOLE-BODY INHALATIONTOXICITY 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 2.8Table A GASOLINE TBA VAPOR CONDENSATE:A 13-WEEKWHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131

Chamber Monitoring Results

Date Exposure Number

Mean Nominal (mglm3)

1830 2090 2070 1990 2010 2150 2110 2080 1950 2020 2160 2020 2020 2190 2090 2090 2140 2200 2070 2130 Cumulative Exposure Record Grou~ HA

# - 2.000 ma/m3

Mean (mglm")

1840 2000 1935 1933 1953 1975 2005 1970 1963 2135 2188 1953 1845 2043 2023 2013 2070 2090 2068 2183 alytical Chamber Concentration Individual MMAD Olm)

2.312

4.144

0.7525

0.8197 Particle Size ztermina

GSD InsTMC (mglm3)

4.69E-03

9.38E-03

2.35E-02

2.61 E-03 Chamber En Mea 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 2.8Table A GASOLINETBA VAPOR CONDENSATE:A 13-WEEKWHOLE-BODY INHALATIONTOXICITY 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 ParticleSize Determinations alytical Chamber Concentration Nominal (mglm3)

1830 2090 2070 1990 2010 2150 2110 2080 1950 2020 2160 2020 2020 2190 2090 2090 2140 2200 2070 2130 2071

88 Temperature Date Exposure Number Mean (mslm3)

1928 2105 2060 1935 1830 2048 2123 2020 2018 1970 2203 1968 2088 2165 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 MeanTableA GASOLINETBA VAPOR CONDENSATE: A 13-WEEKWHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131

Date Exposure Number

21-Aug-02 

## ( 20 Mean S.D. Chamber Monitoring Results CumulativeExposure Record

Group lllA

# - 10,000 mg/m3
Nominal (mg/m3)

101 00 9960 10200 10400 10300 10400 9790 10000 9580 10500 9910 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 3.2Table A GASOLINETEA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATIONTOXICITY 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 9910 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 1Humidity 

## (W 47 48 49 47 48 48 48 46 39 42 41 44 45 45 46 47 48 45 47 48 45.9 2.7Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEKWHOLE-BODY INHALATIONTOXICITY STUDY IN RATS 00-6131 Chamber Monitoring Results CumulativeExposure 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 ITableA 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. CumulativeExposure 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 20180 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

2.51E-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 55Huntingdon 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 1Huntingdon 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 LIMITSHuntingdon 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 differencesHuntingdon 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 II 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 differencesHuntingdon 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 differencesHuntingdon 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 differencesHuntingdon 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 differencesHuntingdon 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: 

### * = pc0.05Huntingdon 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-PRESENTHuntingdon 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-PRESENTHuntingdon 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-PRESENTHuntingdon 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-PRESENTHuntingdon 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-PRESENTHuntingdon 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-PRESENTHuntingdon 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-PRESENTHuntingdon 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-PRESENTHuntingdon 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-PRESENTHuntingdon 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-PRESENTHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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 11Huntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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 FeederHuntingdon 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 5Huntingdon 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.

NHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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 25

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

105 97 91 106 98 97 105 110 8 8 145 SF 92 108 103 93 MEAN S.D.

NSF=Spilled FeederHuntingdon 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.

NHuntingdon 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.

NHuntingdon 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 HISTORYHuntingdon 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 SACRIFICEHuntingdon 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 SACRIFICEHuntingdon 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 ASSESSMENTSHuntingdon 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 DAYHuntingdon 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