## DRAFT 

## REPORT Study Title In Vivo-In Vitro Rat Peripheral Lymphocyte Sister ChromatidExchangeAssay
Test Substance Gasoline TBA Vapor Condensate Author Ramadevi Gudi, Ph.D. Caren M. Brown, M.S. Study Completion Date

(Pending finalreport) Performing Laboratory BioReliance 9630Medical CenterDrive Rockville, Maryland 20850 BioReliance StudyNumber

Client Huntingdon Life Sciences Princeton Research Center (PRC) MettlersRoad East Millstone, New Jersey 08875-2360 HLS StudyNumber -



## Page 

## 1of21

# STATEMENT 

# OF COMPLIANCE
The SisterChromatidExchangeAssay,BioRelianceStudy No. AA40PA.130.BTL,was conducted in compliance with US EPA 79.60, CFR Vol. 59,No. 122,27June 1994and US EPA GLP 79.60, CFR Vo. 59,No. 122,27 June 1997. This study was performed according to protocol and BioReliance's Standard OperatingProcedure for Sister ChromatidExchange Assay with no exceptions. 

# -&. Rarnadevi Gudi, Ph.D. Date Principal Investigator BioReliance 

## Gary

# M. Hoffman,B.A., 

## D.A.B.T. Date StudyDirector HuntingdonLife Sciences

BioReliance Study

## No. : 

### AA40PA. 

## 130.BTL 2

# Quality Assurance Statement

Study Title: 

### IN VIVO-IN VITRO RAT PERIPHERAL LYMPHOCYTE SISTER CHROMATID EXCHANGE ASSAY studyNumber: 

### AA40PA. 130.BTL Study Director: Ramadevi Gudi,Ph.D. This study has been divided into a series of in-process phases. Using arandom sampling approach, Quality Assurance monitors each of these phases over a series of studies. Procedures, documentation, equipment records,etc., are examined in order to assure that the study is performed in accordance with the U.S. FDA Good Laboratory Practice Regulations (21 

### CFR 

## 58), the U.S. EPA GLPs (40 

### CFR 792 and 40 CFR 160), the 

### UK GLP Regulations, the Japanese GLP Standard, and the OECD Principles ofGood Laboratory Practice and to assure that the study is conducted according to the protocol and relevant Standard OperatingProcedures. The following are the inspection dates, phases inspected, and report dates of QA inspections ofthis study.
Inspect On 26-Aug-02 

## - 26-Aug-02To Study Dir 26-Aug-02 To Mgmt 29-Aug-02 Phase Harvest of cells Inspect On 20-Dec-02 - 

### 22-Dec-02 To Study Dir 24-Jan-03To Mgmt 24-Jan-03 Phase Draft Report
This report describesthe methods and procedures used in the study and the reported results accurately reflect the raw data ofthe study.

DATE QUALITYASSURANCE

BioReliance 

### StudyNo. :

### AA40PA. 

### I30.BTL 3

# PRC

# - QUALITY ASSURANCESTATEMENT 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 StudyDirector and Management. Thisreport reflects theraw 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 GenotoxicityDose Prep & 

## Adm Genotoxicity Necropsy 22 Aug 02 22 Aug 02

22 Aug 02

Fran Jannone, 

## B

# . A., RQAP-GLP Quality Assurance SeniorAuditor Date

BioReliance

### StudyNo.: AA4OPA. 130.BTL 4

## SIGNATUREPAGE SCIENTIST Thefollowing Scientistwas 

## responsible for the overallconductof the SCEportion ofthis study:

Ramadevi Gudi, Ph.D. Principal Investigator BioReliance Date



# SCIENTIFICREVIEW The following Scientistshave reviewed and approved this report:

David Jacobson-Kram,Ph.D., DABT Date Vice-president, Toxicology BioReliance 

## Gary

# M. Hoffman, B.A., DABT Date Study Director Huntingdon Life Sciences

BioRelianceStudy

## No. : 

### AA40PA. 130.BTL

## InVivo-In Vitro Rat Peripheral Lymphocyte Sister Chromatid Exchange Assay
DRAFT REPORT
Client:

Study Director: Performing Laboratory (Subcontractor):
BioReliance Principal Investigator: Test Substance

## I.D.: SponsorProjectNumber: BioReliance Study No.: Experimental Start Date for SCE: Experimental Completion Date for SCE: Huntingdon Life Sciences Princeton Research Center (PRC) Mettlers Road EastMillstone, New Jersey 08875-2360 GaryHoffman, B.A., DABT BioReliance 9630 Medical Center Drive Rockville, Maryland 20850 Ramadevi Gudi, Ph.D. Gasoline TBA Vapor Condensate 00-6131 AA40PA.130.BTL 23

## August 2002 21October 2002

BioRelianceStudy

## No. : AA4OPA. 

### 130.BTL

# TABLEOF CONTENTS Page

SUMMARY

# .......................................................................................................................................................... 8 

### PURPOSE 

# ............................................................................................................................................................9 

### CHARACTERIZATION OF TESTAND CONTROL SUBSTANCES

# ..................................................... 9 

### MATERIALSAND METHODS

# ........................................................................................................................ 9 

### RESULTS AND DISCUSSION

# ........................................................................................................................ 11 CONCLUSION

# .................................................................................................................................................. 12 REFERENCES

# .................................................................................................................................................. 13 DATA TABLES 

# ................................................................................................................................................. 14 APPENDIX A: CONTRACTING SPONSOR'S EXPOSUREDATA AND ANIMALDATA

# ................... 21



## BioReliance Study

### No


## .:

### AA4OPA


## . 130.BTL

## SUMMARY
The test substance, Gasoline TBA Vapor Condensate, was tested in the rat peripheral lymphocyte sister chromatid exchange (SCE)assay followinginhalation exposureofrats. Information on test system, experimental design and methodology will be provided by Huntingdon Life Sciences.All in vivo portions of the study, including inhalation exposures were performedby Huntingdon Life Sciences. Rats were exposed by inhalation to negative (air) control, 2,000, 10,000 or 20,000 mg/m3of Gasoline TBA Vapor Condensate for 4 weeks 

## (5 days exposure per week). One additional group of animals (5 males and 5 females) was dosed with 

## 5 mglkg of cyclophosphamide (CP, positive control) by intraperitoneal injection 24 hours prior to blood collection. BioReliancepersonnel collectedrat blood samples at 

## 24 hours after the end of a 

# 4 week (5 days per week) exposure period. Rat blood sampleswere cultured and processed forthe 

## SCE assay atthe BioReliance facility. A minimum of 25 second divisionmetaphases per animal were scored for SCEs. At least 100consecutivemetaphasesper animalwere scored for the number ofcells in first-, second-, or third-division metaphase for each animal 

## as 

## an indictor of toxicity (cell cycle delay). At least 1000 cells were scored for mitotic index per animal. A one-tailed Dunnett's t test for multiple comparisons was performed to compare the SCE frequency of test exposure levels to the negative control frequency. No statistically significant increase (p0.05) in average SCE frequency was observed in males or females in any groups exposed to Gasoline TBA Vapor Condensate. Based onthe findings of this study, the test substance, Gasoline TBA Vapor Condensate, was concluded to be negative for the induction of sister chromatid exchanges in rat peripheral lymphocytes.

BioReliance Study

## No. : 

### AA40PA 130.BTL

## PURPOSE The purpose of this study was to evaluate the potential of Gasoline TBA Vapor Condensateto induce SCEsinrat peripheral lymphocytes following inhalationexposures.


## CHARACTERIZATIONOF TEST AND CONTROL SUBSTANCES Information regarding test substance and negative control and their characterizationswill be providedby Huntingdon Life Sciences. Cyclophosphamide (CP, CAS No. 6055-19-2, lot number 9 1 Kll76, expiration date 30 September 2004, white powder, storage 2-8OC, purity 99.5% by HPLC), was obtained from the Sigma Chemical Company St. Louis, 

# MO (responsible for its characterization). CP was shipped overnight on 19 August 2002 from BioReliance to Huntingdon Life Sciences. CP was dissolved and diluted in sterile distilled water at ~untih~don Life Sciences to a stock concentrationof0.5 mg/mL foruse asthe positive control forthe SCE study. CP has been characterized as per the Certificate of Analysis on file with the testing facility. The stability of CP and of the CP mixtures used in this assay was demonstratedby acceptableresults that met the criteria for a valid test. 

## MATERIALSAND METHODS Test System Information on the test system will be provided by Huntingdon Life Sciences. 

## Sister Chromatid Exchange Assay Peripheral blood lymphocytes were obtained from Sprague-Dawley rats that were exposed with negative (air) control, 2000, 10,000 or 20,000 mg/m3of Gasoline TBA Vapor Condensate, or positive control (5 mgkg of CP injected 

# IP at 10 rnL/kg, 24 hours prior to sacrifice). Animals were anesthetized with isoflurane by inhalation exposure. Blood samples were collectedinto sodium heparin tubes from the abdominal aorta by BioReliancepersonnel on site at Huntingdon Life Sciences. The blood samples were transported to BioReliance on the day ofcollectiononwet ice. 

## Cell Culture and CoIIectionof Metaphase Cells Within 24 hours after blood collection, the whole blood samples were cultured in duplicates per animal in 

# RPMI 1640 culture medium with 25 mM Hepes buffer supplemented, 10% fetal bovine serum, antibiotics (pencillin G, 100dmL and streptomycin sulfate, 0.1 mg/mL), 20 pg/mL phytohemagglutinin and an additional 2 

# mM L-glutamine. Cultures were initiated by inoculating 0.5 mL of whole blood per 

## 5 

# mL of complete medium
BioReliance Study

### No. :

### AA40PA. 130.BTL 9

## in T-25 sterile disposable tissue culture flasks and incubated at 37OC. Approximately 21 hours after initiation of the cultures, the cells were exposed to 5 &mL of bromodeoxyuridine (BrdU, Sigma Co., St. Louis, MO). Approximately 68 hours after initiation of the cultures, 0.2 pg/dof colcemid was added to each flask and incubated for approximately 4 hours. Approximately 72 hours after initiationofthe cultures,the cells were collected by centrifugation at approximately 1200 rpm for about 5 minutes. The cell pellet was resuspended in 5 mL 0.075 M KC1 and incubated at 37f l°Cfor 20 minutes. At the end of the KC1 treatment and immediately prior to centrifuging, the cells were gently mixed and approximately 0.5 mL 

## of fixative (methano1:glacial acetic acid, 3:l vlv) was added to each tube. The cells were collectedby centrifugation, the supernatant aspirated, and the cells were fixedwith two washes with approximately 3-5 

# mL of fixative and stored in fixativeovernight or longer at approximately24°C. 

## SlidePreparation To prepare slides, the fixed cells were centrifuged at approximately 1200 rpm for 5 minutes, the supernatant was aspirated, and the cells were resuspended in 1 

# rnL fresh fixative. The cells were collected by centrifugation and the supernatant aspirated, leaving 0.1 to 0.3 mL fixative above the cell pellet. One to two drops of the cell suspension was dropped onto a glass slide and allowed to air dry overnight. Slides were identified by the studynumber, animal number, replicate tube designation and dateprepared. The dried slides were then stained. 

## Slide Staining The slides were stained by using a modified flourescence-plus-Giemsa 

## technique. Hoechst 33258 stain, 5 pg/mL, was prepared freshby diluting40 mL 

## ofHoechst 33258 stock stain 50 pg/mL in distilled water, with 360 mL 

## phosphate buffer, pH 6.8. The slides were stained in Hoechst, 5 pg/mL, for 10 minutes, rinsed in deionized water and dried between sheets of bibulous paper. Three drops of phosphate buffer was placed on each slide and the buffer covered with a coverslip. The slides were then placed on a slide warmer maintained at approximately 60-+2OC and covered with a "Black Ray" lamp with 15 watt tubes for 

# 4-6 minutes. The coverslips were removed, the slides rinsed in deionized water and counterstained with 5% Giemsa solution (fieshly prepared by diluting 

## 20 mL Giemsa in 380 mL deionized water) for 6-10 minutes. The slides were rinsed in deionized water and leftto air dry. 

## Evaluation ofSCEs Slideswere coded using random numbers by an individual not involved with the scoring process. Metaphase cells were examined under oil immersion without prior knowledge of treatment groups. Whenever possible, a minimum of 

# 25 second division metaphases per animal were scored for SCEs. At least 100 consecutive metaphase cells were scored for the number of cells in first-, second- or third-division metaphase for each animal as an indicator oftoxicity (cell cycle delay). At least 1000cells were scored for mitotic indexper animal.

BioReliance Study 

## No. :

### AA40PA. 130.BTL

## Evaluation ofTest Results The number of SCEs per second-divisionmetaphase cell were presented for each group along with the range of SCEsImetaphasefor animals within a group. SCE frequenciesas the grand mean 

# * standard deviation were presented for each group. The Average Generation Time (AGT) is estimated as: = 

## Number of hours 

## in BrdU 

# X 1001 [(number 

# M, cells x l)+(nurnber 

# M, cellsx 2)+(number 

# M, cellsx 

## 3)]. A Dunnett's t test for multiple comparisons was performed to compare the SCE frequency of test exposure levels to the negative control frequency, and if p50.05, that exposure level was considered significant. The conclusion of the study was based on the Principal Investigator's evaluation of all the data, including the biological as well as statistical significance. The following criteria were used as guidelines 

## in evaluationofthis study: The test substance was considered positive if an exposure-level responsive and statistically significant increase is observed over a minimum of two exposure levels. A statistically significant increase at the high exposure level with an exposure-levelresponsive although not statistically significant increase at lower exposure levels was assessed 

## as suspect. A statistically significant increase at one or more exposure levels with no evidence of an exposure level response was assessed as 

## equivocal or as negative according to the magnitude ofthe response and the number ofexposure levels affected. CriteriaforDeterminationofa Valid Test The mean SCEs/cell/animal for the positive control animals must be statistically increasedrelativeto the negative control. Deviations No known deviations from the protocol or assay-method SOPS occurred during the conduct of this study. Archives All raw data, the protocol and all reports generated at BioReliance will be maintained according to Standard Operating Procedure OPQP3040 by the BioReliance RAQA unit headquartered at: BioReliance, 14920 Broschart Road, Rockville, 

# MD 20850. After submission ofthe final report, all of the above will be shipped to Huntingdon Life Sciences to be archived. All study materials returned to Huntingdon Life Scienceswill first be copied and the copy will be retained in the BioReliance archives for a minimum of 10 years. All specimens, such as microscope slides, will be held in storage as long as the quality affords evaluationat BioRelianceuntil the final report isissued in accordance with the relevant Good Laboratory Practice Regulations. RESULTS

# AND DISCUSSION Sister chromatid exchanges involve a symmetricalexchange at one locus,between sister chromatids of the chromosomes. Giemsa method for the differential staining of sister chromatids by Perry and Wolff (1974a) Wolff and Perry (1974) have made it possible to BioReliance Study

### No. :AA40PA. 

### 130.BTL 11

## distinguish between sister chromatids. These techniques involve exposing cells to 5-bromodeoxyuridine (BrdU) for at least 2 rounds of replication so that M2 chromosomes consist of one chromatid unifilarly substituted with BrdU and the other bifilarly substituted. The chromatids of such chromosomes stain differentially with Giemsa stain. The observed SCE represents a recombination between DNA helices in the replicating chromosomes and used as a sensitive cytological end point which correlates with DNA damage. The test substance, Gasoline TBA Vapor Condensate, was tested in the rat peripheral lymphocyte sister chromatid exchange (SCE) assay following inhalation exposureofrats. Information on test system, experimental design and methodology will be provided by Huntingdon Life Sciences. All in vivo portions of the study, including inhalation exposures, were performed by Huntingdon Life Sciences. Rats were exposedby inhalationto negative (air) control, 2,000, 10,000or 20,000mg/m3 of Gasoline TBA Vapor Condensate for 4 weeks (5 days exposure per week). One additional group of animals (5 males and 5 females) was dosed with 5 mgkg cyclophosphamide(CP, positive control) by intraperitoneal injection 24 hours prior to blood collection. Animals dosed at 5 mgkg of CP were used as the positive control group for the SCE assay. BioReliance personnel collected rat blood samples at 24 hours after the end of a 4 week (5 daysper week) exposureperiod. Rat blood samples were cultured and processed for the SCE assay at BioReliance. A minimum of 25 second division metaphasesper animalwere scored for SCEs. At least 100 consecutive metaphasesper animal were scored for the number of cells in first-, second-, or third-division metaphase for each animal as an indictor of toxicity (cell cycle delay). At least 1000 cells are scored for mitotic index per animal. The number of SCEs per second-division metaphase cell were presented for each animal (both males and females) in Tables 1 to 5. The number of MI, M2 and 

## M3 cells out of 100 metaphases and the mitotic index out of 1000 cells per animal are also presented in Tables 1 to 

## 5. The range and the average SCEs per animal and the SCE frequencies as the grand mean 

# * standard deviation per group for males are presented in Table 

# 6 and for females in Table 7. The Average GenerationTime (AGT) andthe mitotic indexper groupfor males arepresented in Table 

## 6 and for females in Table 

## 7. A Dunnett's t test for multiple comparisons was performed to compare the average SCE frequency of test exposure levels to the negative control frequency. No statistically significant increase in average SCE frequency (p>0.05) was observed in males or females in any groups exposed to Gasoline TBA Vapor Condensate. The positive control group (CP, 5 mg/kg) induced statistically significant increases in SCE frequency (p10.05) in both males and females. No appreciable difference was observed in AGT and in mitotic index in the test substance groups relativeto the negative control group. 

# CONCLUSION The positive control fulfilledtherequirements for a valid test. Under the conditions of the assay described in this report, based on the findings of this study,the test substance, Gasoline

# TBA Vapor condensate, the induction of sister chromatid exchanges

# inrat peripheral was concluded to be negative for lymphocytes.

BioRelianceStudy 

## No. :

### AA4OPA. 130.BTL

## REFERENCES Evans,

## H.J. (1976) Cytological methods for detectingchemical mutagens,in: 

## A. Hollaender (Ed.), ChemicalMutagens, Principles and Methods fortheirDetection, vol. 4. Plenum Press, New York, NY. MINITAB User's guide 2: data Analysis and Quality Tools, Release 12for Windowsm 95 WindowsNTm February 1998Chapter 3: Analysis ofVariance, 3-1 to3-7. PenyP. and Wolff 

## S. (1974a)New Giemsamethod for the differential staining of sister chromatid, Nature (London)

## 251, 1 56-

## 1 58 Wolff 

## S. and Peny P. (1974) Differential Giemsa stainingof sister chromatid andthe study of sister chromatidexchanges without autoradiography. Chromosoma48,341-353

BioReliance Study

## No. :

### AA40PA. 130.BTL

### TABLE 1 

## Number of Sister Chromatid Exchanges per cell in Negative Air Control Group Air only Male and Female Rats 

# 1 Number of 

## SCE's Counted 

# I I I Animal Number 

# I



# 'Animal numbers 1081-1085are males and 1591-1595are females.

Number ofMI,

## M2 and M3 cells are obtained from 100 metaphases. Mitotic index; number mitotic figures x 100/1000cellscountedkmimal.

BioRelianceStudyNo. :

## AAIOPA. 130.BTL 14

### TABLE2


### Number of Sister Chromatid Exchanges per cell at 2000 mg/m30f Gasoline 

### TBA Vapor Condensate Male and Female Rats 

# 1 Number 

## ofSCE's Counted 

# I I I Animal Number 

# 1



# 'Animal numbers 2071-2075are males and 2581-2585 are females.

Number ofM1, M2 and 

## M3 cells are obtained fiom 100metaphases. Mitotic index; number mitotic figures 

## x 100/1000cells counted/animal.

BioReliance Study No. :

### AA40PA. 130.BTL

### TABLE 3 

## Number of Sister Chromatid Exchanges per cell at 10,000 

## rng/m3of GasolineTBA

## Vapor Condensate Male and Female Rats



# 'Animal numbers3071-3075 are males and3581-3585 are females.



## NumberofMI, 

## M2 andM3 cells are obtainedfrom 100metaphases. Mitoticindex;numbermitotic figures

## x 100/1000cellscountedlanirnal.

BioReliance StudyNo. :

### AA4OPA. 

## 130.BTL

### TABLE 4 

### Number 

## of Sister Chromatid Exchanges per cell at 

## 20,000 mg/m3 

## of Gasoline 

### TBA Vapor Condensate Male and Female Rats 

# I Number

## of SCE's Counted 

# I

'Animal numbers 4081-4085 are malesand4591-4595 are females.

Numberof

## MI,

## M2 andM3 cells are obtainedfrom 100metaphases. Mitotic index;numbermitotic figuresx 100/1000cells countedlanimal.

BioReliance StudyNo. : 

## AA40PA. 130.BTL

### TABLE 5 Number of Sister Chromatid Exchanges per cell at 

### 5 mglkg Cyclophosphamide Male 

### and Female Rats 

## Number of 

## SCE's Counted 

# I AnimalNumber 

## CellNo. 

## 5051 5052 5053 25 20 28 20 21 20 21 23 25 

## 4 22 24 26



# 'Animal numbers 5051-5055arema11 ;

## and 5561-5565 are females.

Number of

## M1,M2 

## and M3 cells are obtained from 100metaphases. Mitotic index; number mitotic figures

## x 100/1000cells counterYanimal.

BioRelianceStudyNo. : 

### AA40PA. 

## 130.BTL 18

### TABLE 6 

### Summary of Sister Chromatid Exchange Data -Gasoline TBA Vapor Condensate MaleRats Sister Chromatid Exchange data Total Range of # 

### of # 

### of SCEs Mean Group Group Treatment Animal Metaphases SCEsper per Animal SCEs Range Mean # 

### Scored Animal Min Max per cell Min Max Std. Dev.* Air 1081 25 1082 25 1083 25 1084 25 1085 25 

### Gasoline TBA Vapor Condensate 2000 

### mg/m3 2071 25 2072 25 2073 25 2074 25 2075 25 

### GasolineTBA Vapor Condensate 10,000

### mg/m3 3071 25 3072 25 3073 25 3074 25 3075 25 

### GasolineTBA Vapor Condensate 20,000

### mg/m3 408 1 25 4082 25 4083 25 4084 25 4085 25 

### Cyclophospharnide 5 

### mgfkg 5051 25 5052 25 5053 25 5054 25 5055 25 

### AGT MI per per per 4nimal Group Group



# * The standard deviation

### was calculated using the data of all 125

### metaphases scored. 

## $ =

### ~hnett's t test (p <0.05) 

### Mitotic Index: (Number of cells 

## in mitosis out of 1000

### cells) AGT=Average generationtime: Number of hours in BrdU

## X 1001

### (number M, cells

## x I)+(number 

## M, cells

## x 2)+(number M3cells

## x 3)



### BioReliance Study No. :

### AA40PA. 130.BTL 19

### TABLE 7 

### Summaryof Sister Chromatid Exchange Data -

### Gasoline TBA Vapor CondensateFemale Rats 

### Sister Chromatid Exchange data Total Range of # 

### of # 

### of SCEs Mean Group Group Treatment Animal Metaphases SCEsper per Animal SCEs Range Mean

Air 1591 25 1592 25 1593 25 1594 25 1595 25 Gasoline TBA Vapor Condensate 2000 mg/m3 2581 25 2582 25 2583 25 2584 25 2585 25 Gasoline TBA Vapor Condensate 10,000mg/m3 3581 25 3582 25 3583 25 3584 25 3585 25 Gasoline TBA Vapor Condensate 20,000 mg/m3 459 1 25 4592 25 4593 25 4594 25 4595 25 AGT 

## - MI per per per # 

### Scored Animal Min Max per cell Min Max + 

### Std. Dev.*

Cyclophospharnide 5mgk 5561 25 598 19 

# - 31 23.9 5562 25 496 15 

## - 26 19.8 5563 25 494 14 

# - 24 19.8 14 to 31 22.1* 

# * 3.4 5564 25 614 20 

# - 31 24.6 5565 25 558 20 

# - 31 22.3 Animal Group Group



# * The standard deviation

## was calculated using the data ofall 125metaphases scored. 

# 1

=

### Dunnett's t

### test 

## (p 4.05) Mitotic Index: (Number

## of cells in mitosis out of 1000cells) AGT=Averagegenerationtime: Number ofhours 

## in BrdU

## X 100/(numberM, cells

## x l)+(number M, cells x 2)+(numberM, cells

## x 3)

BioReliance StudyNo. :

### AA40PA. 130.BTL 20

### AppendixA: 

### Contracting Sponsor's Exposure Data and Animal Data



### BioReliance StudyNo. :

### AA40PA. 130.BTL 2

### 1

## Huntingdon Life Sciences 00-6131 Page 1 21 1-TBA-S Draft Final Report



# INTRODUCTION: The following is data generated at Huntingdon Life Sciences,East Millstone,NJ. The separatelyissued main studyreport should be referenced for details of the procedures used for test atmosphere 

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



# STUDY DATES: Date of Animal Receipt: 15July 2002 Appendix A

Experimental Initiation Date: 26 July 2002 (in-life) Experimental Completion Date: 22 August 2002 (in-life) Draft Report Date: 2 June 2004


# 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. Summaryof Clinical Observations(pretest only)

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

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

# ........................................................ E. Mean Feed ConsumptionValues (grams/kg/day)

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

# ............................................................... G. 

## IndividualBody Weights (grams)

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

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

## I. IndividualFeed ConsumptionValues (gramskglday)


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



# ............................................................................................ J. Animal TerminationHistoryAppendix A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131



### Chamber 

### Monitoring Results Cumulative Exposure RecordAppendix A GASOLINETBA VAPOR CONDENSATE: A 13-WEEKWHOLE-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 IB

# - 0 

### (air only) mg/m3
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 (mglm3) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 

### Individual MMAD Olm)

3.102

2.850

0.7383 

### Particle Size nsTMC

Owm3)

7.27E-03

5.51E-03

2.12E-02 

### Chamber Environment Mean Temperature ("C) 23 24 24 25 25 25 25 25 24 24 24 24 24 25 25 25 25 25 24 23 

### 24.4 0.7 

### Humidity ("@ 55 55 53 53 56 56 56 53 45 49 50 52 53 54 54 56 54 52 56 56 

### 53.4 2.8Appendix A GASOLINETBA VAPOR CONDENSATE: A 13-WEEKWHOLE-BODY INHALATION TOXICITY STUDY 

### IN RATS 00-6131



### Chamber Monitoring Results Cumulative Exposure Record Group IIA

# - 2,000 mglm3
Date Particle Size Chamber Environment Mean Nominal (mglm")

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

### alytical Chamber Concentration Individual msTMC (mglm")

4.69E-03

9.38E-03

2.35E-02 

### Humidity 

## ("4 50 50 48 49 5 1 51 50 48 40 44 44 46 47 47 48 50 49 48 48 52 

### Exposure Number Temperature ("C) 23 23 24 23 23 23 23 23 24 24 24 24 24 23 23 23 23 23 24 24 

### Mean (mglmg)

1840 2000 1935 1933 1953 1975 2005 1 970 1963 2135 2188 1953 1845 2043 2023 201 3 2070 2090 2068 2183 

### MMAD Olm)

2.312

4.144

0.7525 

### eterrnina GSD

MeanAppendix A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEKWHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131

Chamber Monitoring Results

Date Cumulative Exposure Record Group IIB

# - 2,000 mg/m3 

# , Number

Mean 

# 1 2071 S.D. 

# 1 88 Mean (mg/m3)

1928 2105 2060 1935 1830 2048 2123 2020 2018 1970 2203 1968 2088 2165 2008 1995 2028 2073 2023 2025 alytlcal Chamber Concentration Individual I MMAD Olm)

3.582

2.398

0.7703 Particle Size !terminal

GSD nsTMC mm3)

6.02E-03

2.09E-03

1.73E-02 Chamber Environment Mean Temperature ("C) 22 22 23 23 23 23 23 23 23 23 23 22 23 24 23 23 23 24 22 22 22.9 0.6 Humidity 

# ("w 52 53 50 52 54 55 55 5 1 43 46 45 50 49 50 52 53 53 50 54 56 51.2 3.4Table A GASOLINE TBA VAPOR CONDENSATE:A 13-WEEK WHOLE-BODY INHALATIONTOXICITY STUDY IN RATS 00-6131
Chamber Monitoring Results Cumulative Exposure Record

Grou~ IIIA 

# - 10.000 ma/m3 Chamber Environment Particle Size ~alytical Chamber Concentration Individual Nominal (mglm")

10100 9960 10200 10400 10300 10400 9790 10000 9580 10500 9910 9770 9770 10000 9560 9660 9850 10100 10600 10000 10020 302 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 Date Exposure Number Humidity 

## ("4 50 50 50 50 52 52 52 49 40 44 44 47 47 49 51 52 52 48 50 51 49.0 3.2 1 Mean (mdm3)

10490 9908 10130 lo380 10780 10000 10500 9818 10210 lo580 10200 10010 9763 10190 10220 9463 10180 10020 10440 9868 erminati GSD MMAD Olm)

8.779

2.937

0.7457

0.8931 TMC (mdm")

7.41 E-03

3.72E-03

2.45E-02

2.23E-03 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Mean S.D.Table A GASOLINETBA VAPOR CONDENSATE: A 13-WEEKWHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131



### Chamber Monitoring Results Cumulative Exposure Record

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. Group lllB

# - 10,000mg/m3 

# I Particle Size Nominal 

## 1 Analvtical Chamber Concentration Determinations Individual (mg/m3) 10100 10100 10100 11000 11200 9920 10600 10100 10100 10400 10100 10100 10000 10500 10700 10900 11100 11000 11100 10600 11800 11300 9950 10400 9920 1 1000 10300 10200 9420 1 1200 9200 8420 10300 10000 8810 10500 9800 10600 9320 10500



### lo260

588 (mslm3)

10100 9960 10200 10400 10300 10400 9790 10000 9580 10500 991 0 9770 9770 10000 9560 9660 9850 10100 10600 10000 

### 10020

302 TMC (mg/m3)

2.32E-03

2.80E-03

2.68E-02

3.03E-03 

### Mean (mglm3)

10010 10500 10020 10000 10090 10160 101 00 10240 10620 10750 11150 10260 10680 10280 10240 9605 10330 9880 10380 9955 

### Chamber Environment Mean Temperature ("C)

23 

### 23.8 0.4 Humidity 

### ("A)

48 

### 45.9 2.7

### Appendix A GASOLINETBA VAPOR CONDENSATE: A 13-WEEK 

### WHOLE-BODY INHALATIONTOXICITY STUDY IN RATS 00-6131



### Chamber Monitoring Results Cumulative Exposure Record

Grou~ IVA

# - 20,000 

### ms/m3
Date Exposure Number 1 2 3 4 

### 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 

### Mean S.D. Nominal (mglm3)

18900 19100 18400 18500 18900 191 00 18900 18800 191 00 19700 18800 18700 18800 19000 19500 19400 19100 19300 19200 18800 19000 321 

### Mean (mslm3) 20500 20650 19800 20380 19500 19480 19250 19530 19950 21380 20530 19480 19200 18500 19000 19430 20630 19500 20250 20300 

### lndivldual MMAD Olm)

1.689

2.505

0.7685 

### Particle Size !terminat

GSD 

### TMC (mslm3)

2.75E-03

3.68E-03

1.81E-02 

### Chamber 

### En Mea Temperature ("C) 26 26 26 25 25 25 25 25 26 26 26 26 26 25 25 25 25 25 26 26 25.5 0.5 

### ronment Humidity 

## ("/.I 5 1 52 52 51 52 52 53 5 1 43 45 46 49 49 50 53 55 53 50 51 54 50.6 3.0Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEKWHOLE-BODY INHALATIONTOXICITY STUDY IN RATS 00-6131

Date Exposure Number

-

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Mean S.D. Chamber Monitoring Results CumulativeExposure Record

Group IVB

# - 20,000 mg/m3
Nominal (mg/m3)

18900 19100 18400 18500 18900 191 00 18900 18800 19100 19700 18800 18700 18800 19000 19500 19400 19100 19300 19200 18800 19000 321 1 Mean (mg/m3)

19550 19650 20080 19400 20250 201 80 19600 19880 19880 20650 19330 20300 20050 1 9750 19980 20580 20650 20330 20030 20500 ~alytical Chamber Concentration Individual I MMAD bm)

1.032

1.793

0.7982

0.9155 

## I Chamber Environment Particle

### Size 

## I MeanHuntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TBA-S PAGE 1
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 MALES SUMMARY OF CLINICAL OBSERVATIONS

NORMAL WITHIN NORMAL LIMITSHuntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TEA-S 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 5 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 MALES MEAN BODY WEIGHTS (GRAMS)

---------------------------------------------------------------------------------------------------------------------------..------ DOSE GROUP: I I I I11 I V V DOSE LEVEL (MG/M3 ) : 0 2000 10000 20000 SCE+CONTROL



### ................................................................................................................................... DAY -3 MEAN S.D.

N DAY 4 MEAN S.D.

N MEAN S.D.

N DAY 11

DAY 18 MEAN S.D.



### N DAY 25 MEAN 355 349 350 S.D. 30.9 22.2 26.8

N 5 5 5



### ......................................................................................... 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 I I I11 I V V DOSE LEVEL (MG/M3): 0 2000 10000 20000 SCEtCONTROL

DAY -3 MEAN S.D.

N 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 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: DOSE LEVEL (MG/M3) : 

### - - .................... DAY -3 TO 4 MEAN S.D.

N DAY -3T0 11 MEAN S.D.

N DAY -3 TO 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 -3 TO 4 MEAN S.D.

N DAY -3 TO 11 MEAN S.D.

N DAY -3T0 18 MEAN S.D.

N DAY -3T0 25 MEAN 107 94 S.D. 16.8 20.8

N 5 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 I I I I I I V V DOSE LEVEL (MG/M3): 0 2000 10000 20000 SCEcCONTROL

-------------------------.--------------------------------------------------------------------------------------------------------- DAY 4

DAY 11

DAY 18 MEAN S.D.

N MEAN S.D.

N MEAN S.D.

N DAY 25 MEAN 85 8 8 83 83 84 S.D. 3.8 3.7 3.7 1.8 4.0

N 5 5 5 5 5



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



### ................................................................................................................................... DOSE GROUP: I I I III I V V DOSE LEVEL (MG/M3) : 0 2000 10000 20000 SCE+CONTROL

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 SPONSOR STUDY NO. 211-TEA-S TABLE 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

.

### -
ANIMAL# OBSERVATIONS
1081 WITHIN NORMAL LIMITS 1082 WITHIN NORMAL LIMITS 1083 WITHIN NORMAL LIMITS 1084 WITHIN NORMAL LIMITS 1085 WITHIN NORMAL LIMITS



### ................................................ CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT PAGE 1Huntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TBA-S TABLE 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 CLINICAL OBSERVATIONS MALES GROUP 2 2000 MG/M3

-------------------------------------------------------------------------.--.---------..------.------------------------- DAY OF ANIMAL# OBSERVATIONS STUDY 4

-----------------------------------------------------------.-------- 2071 WITHIN NORMAL LIMITS P 2072 WITHIN NORMAL LIMITS P 2073 WITHIN NORMAL LIMITS P 2074 WITHIN NORMAL LIMITS P 2075 WITHIN NORMAL LIMITS P



### .................................................................... CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT PAGE 2Huntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TBA-S PAGE 3 TABLE 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

ANIMAL# OBSERVATIONS DAY OF STUDY 4
3071 WITHIN NORMAL LIMITS P 3072 WITHIN NORMAL LIMITS P 3073 WITHIN NORMAL LIMITS 3074 WITHIN NORMAL LIMITS P 3075 WITHIN NORMAL LIMITS P



### ................................................................................................................................... CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENTHuntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TBA-S TABLE F GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK

MALES GROUP 4 20000 MG/M3



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



### ................................... 4081 WITHIN NORMAL LIMITS 4082 WITHIN NORMAL LIMITS 4083 WITHIN NORMAL LIMITS 4084 WITHIN NORMAL LIMITS 4085 WITHIN NORMAL LIMITS IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS INDIVIDUAL CLINICAL OBSERVATIONS PAGE 4

------------------------------------------------------------------------------.--------------------------------------..------------ CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENTHuntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TBA-S

GASOLINE TOXICITY IN PAGE 5 TABLE F TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS INDIVIDUAL CLINICAL OBSERVATIONS

OBSERVATIONS

.---------------------- WITHIN NORMAL LIMITS WITHIN NORMAL LIMITS WITHIN NORMAL LIMITS WITHIN NORMAL LIMITS WITHIN NORMAL LIMITS DAY OF STUDY 4



### ................................................... CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENTHuntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TBA-S PAGE 6
TABLE 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 CLINICAL OBSERVATIONS FEMALES GROUP 1 0 MG/M3

--------------------------------------------------------------. DAY OF ANIMAL# OBSERVATIONS STUDY

-----------.---------------------------------------------- 1591 WITHIN NORMAL LIMITS 1592 WITHIN NORMAL LIMITS 1593 WITHIN NORMAL LIMITS 1594 WITHIN NORMAL LIMITS 1595 WITHIN NORMAL LIMITS

---------------.------------------------------------------ CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENTHuntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TEA-S PAGE 7 TABLE F 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 CLINICAL OBSERVATIONS FEMALES GROUP 2 2000 MG/M3

ANIMAL# OBSERVATIONS



### ................................................ 2581 WITHIN NORMAL LIMITS 2582 WITHIN NORMAL LIMITS 2583 WITHIN NORMAL LIMITS 2584 WITHIN NORMAL LIMITS 2585 WITHIN NORMAL LIMITS



### ................................................ CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENTHuntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TBA-S

GASOLINE TOXICITY IN
FEMALES GROUP 3 10000 MG/M3 PAGE 8 TABLE F TEA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS INDIVIDUAL CLINICAL OBSERVATIONS

3581 WITHIN NORMAL LIMITS 3582 WITHIN NORMAL LIMITS 3583 WITHIN NORMAL LIMITS 3584 WITHIN NORMAL LIMITS 3585 WITHIN NORMAL LIMITS

---------------.---------------------------------- CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENTHuntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TBA-S TABLE 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 CLINICAL OBSERVATIONS

DAY OF ANIMAL# OBSERVATIONS STUDY 4



### ..................................................................... 4591 WITHIN NORMAL LIMITS P 4592 WITHIN NORMAL LIMITS P 4593 WITHIN NORMAL LIMITS 4594 WITHIN NORMAL LIMITS P 4595 WITHIN NORMAL LIMITS P



### ..................................................................... CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT PAGE 9Huntingdon Life Sciences 00-6131G SPONSOR STUDY NO. 211-TBA-S TABLE 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 CLINICAL OBSERVATIONS

DAY OF ANIMAL# OBSERVATIONS STUDY



### .......................................................... 5561 WITHIN NORMAL LIMITS 5562 WITHIN NORMAL LIMITS 5563 WITHIN NORMAL LIMITS 5564 WITHIN NORMAL LIMITS 5565 WITHIN NORMAL LIMITS



### .......................................................... CODE: 1-SLIGHT 2-MODERATE 3-MARKED P-PRESENT PAGE 10Huntingdon 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 INDIVIDUAL BODY WEIGHTS (GRAMS) MALES GROUP I 0 MG/M3

----------.------------------------------------------------------------------------------------------------------------------------ DAY OF STUDY ANIMAL# 

### - 3 4 11 18 2 5 1081 143 200 250 304 342 1082 141 197 252 309 357 1083 155 211 263 321 363 1084 167 230 290 349 398 1085 132 181 229 275 314 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

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

----------------------------------------------------------.--------------------------.--------.------------------------------------ DAY ANIMAL# 

### - 3



### ....................... 4081 143 4082 138 4083 147 4084 162 4085 141 OF STUDY 4 11 18 25 210 262 323 362 186 233 275 312 222 285 343 389 225 289 350 390 205 266 319 362 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 V SCE+CONTROL

------------------------------------------------------------------.----.----------------------------------------------------------- DAY OF STUDY ANIMAL# -3 4 11 18 25

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

-------------------------------------------------.--------------------------------------------------------------------------------- DAY OF STUDY ANIMAL# 

## - 3 4 11 18 25

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 2581 133 159 184 202 215 2582 139 162 193 212 222 2583 117 140 158 182 194 2584 126 161 186 215 226 2585 127 168 197 232 255
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 IMMLTNOTOXICITY ASSESSMENTS

MEAN S.D.

NHuntingdon 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 IMMUNOTOXICITY ASSESSMENTS INDIVIDUAL BODY WEIGHTS (GRAMS) FEMALES GROUP IV 20000 MG/M~

---------------------------------------------------------------------------.------------------------------------------------------- DAY OF STUDY ANIMAL# 

### - 3 4 11 18 25

MEAN S.D.

NHuntingdon Life Sciences 00-6131G Genotoxicity Sub-Group PAGE 10
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 V SCE+CONTROL
DAY OF STUDY ANIMAL# -3 4 11 18 25

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

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 IMMUNOTOXICITY ASSESSMENTS INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) MALES GROUP I 0 MG/M3 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 49 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

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 INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) MALES GROUP I11 10000 MG/M3

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



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

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 (-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) MALES GROUP V SCE+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 INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) FEMALES GROUP I 0 MG/M~

--------------------------------------------------------------------------------------.-------------------------------------------- DAY OF STUDY ANIMAL# -3-4 -3-11 -3-18 -3-25

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 INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) FEMALES GROUP I1 2000 MG/M3

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

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 DAY OF STUDY ANIMAL# -3-4 -3-11 -3-18 -3-25

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 INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) FEMALES GROUP IV 20000 MG/M3



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

MEAN S.D.

NHuntingdon Life Sciences 00-6131G Genotoxicity Sub-Group 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 PAGE 10Huntingdon 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

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

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

MEAN S.D.

NSF=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

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 V SCE+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

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

MEAN S.D.

N OF STUDY 11 18 ---------------

105 97 106 98 105 110 145 SF 108 103

SF=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 ANIMAL# 4



### ....................... 4591 119 4592 119 4593 124 4594 123 4595 127 OF STUDY 11 18 2 5

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

104 9 2 88 101 9 4 89 107 101 93 103 101 95 111 104 9 3 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 V SCE+CONTROL



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

MEAN S.D.

N DAY OF STUDY 4 11 18

SF=Spilled FeederHuntingdon Life Sciences 00-6131G Genotoxicity Sub-Group PAGE 1 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 I 0 MG/M~

................................................................................................................................... TYPE OF DATE OF WEEK OF STUDY ANIMAL# DEATH DEATH STUDY DAY

................................................................................................................................... 1081 TERMINAL SACRIFICE 22-AUG-02 3 27 1082 TERMINAL SACRIFICE 22-AUG-02 3 27 1083 TERMINAL SACRIFICE 22-AUG-02 3 27 1084 TERMINAL SACRIFICE 22-AUG-02 3 2 7 1085 TERMINAL SACRIFICE 22-AUG-02 3 27Huntingdon 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

2071 TERMINAL SACRIFICE 22-AUG-02 2072 TERMINAL SACRIFICE 22-AUG-02 2073 TERMINAL SACRIFICE 22-AUG-02 2074 TERMINAL SACRIFICE 22-AUG-02 2075 TERMINAL SACRIFICE 22-AUG-02Huntingdon 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

TYPE OF DATE OF WEEK OF STUDY ANIMAL# DEATH DEATH STUDY DAY

................................................................................................................................... 3071 TERMINAL SACRIFICE 22-AUG-02 3 27 3072 TERMINAL SACRIFICE 22-AUG-02 3 27 3073 TERMINAL SACRIFICE 22-AUG-02 3 2 7 3074 TERMINAL SACRIFICE 22-AUG-02 3 2 7 3075 TERMINAL SACRIFICE 22-AUG-02 3 27

...................................................................................................................................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 ANIMAL TERMINATION HISTORY MALES GROUP IV 20000 MG/M3

TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICEHuntingdon Life Sciences 00-6131G Genotoxicity Sub-Group PAGE 5 TABLE J GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND I-WEEK IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS ANIMAL TERMINATION HISTORY MALES GROUP V SCE+CONTROL 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 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 ASSESSMENTSHuntingdon 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

.------------------------------------------------------------------------..-----------------------------------.-------------------- 2581 TERMINAL SACRIFICE 22-AUG-02 3 27 2582 TERMINAL SACRIFICE 22-AUG-02 3 27 2583 TERMINAL SACRIFICE 22-AUG-02 3 27 2584 TERMINAL SACRIFICE 22-AUG-02 3 27 2585 TERMINAL SACRIFICE 22-AUG-02 3 27

------------------------------------------------------------------------------.----------------------------------------------------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/M3 TYPE OF DATE OF WEEK OF STUDY ANIMAL# DEATH DEATH STUDY DAY

-----------------------------------------------------------.-----.------------------------.----------------------------------..----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 ASSESSMENTSHuntingdon Life Sciences 00-6131G Genotoxicity Sub-Group

GASOLINE TOXICITY

FEMALES GROUP V SCE+CONTROL



### .................................... TYPE OF ANIMAL# DEATH PAGE 10
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



### .............................................................................. DATE OF WEEK OF STUDY DEATH STUDY DAY



### ................................................................................................................................... 5561 TERMINAL SACRIFICE 22-AUG-02 3 2 7 5562 TERMINAL SACRIFICE 22-AUG-02 3 2 7 5563 TERMINAL SACRIFICE 22-AUG-02 3 27 5564 TERMINAL SACRIFICE 22-AUG-02 3 27 5565 TERMINAL SACRIFICE 22-AUG-02 3 27

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