DRAFT REPORT 

Study Title 

In Vivo-In Vitro Rat Peripheral Lymphocyte Sister Chromatid Exchange Assay 

Test Substance 

Gasoline TBA Vapor Condensate 

Author 

Ramadevi Gudi, Ph.D. 

Caren M. Brown, M.S. 

Study Completion Date 

(Pending final report) 

Performing Laboratory 

BioReliance 

9630 Medical Center Drive 

Rockville, Maryland 20850 

BioReliance Study Number 

Client 

Huntingdon Life Sciences 

Princeton Research Center (PRC) 

Mettlers Road 

East Millstone, New Jersey 08875-2360 

- HLS Study Number 

Page 1 of 21 
STATEMENT OF COMPLIANCE 

The Sister Chromatid Exchange Assay, BioReliance Study No. AA40PA. 130.BTL, was 

conducted in compliance with US EPA 79.60, CFR Vol. 59, No. 122,27 June 1994 and US 

EPA GLP 79.60, CFR Vo. 59, No. 122, 27 June 1997. This study was performed according 

to protocol and BioReliance's Standard Operating Procedure for Sister Chromatid Exchange 

Assay with no exceptions. 

-&. 

Rarnadevi Gudi, Ph.D. Date 

Principal Investigator 

BioReliance 

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

Study Director 

Huntingdon Life Sciences 

BioReliance Study No. : AA40PA. 130.BTL 2 
Quality Assurance Statement 

Study Title: IN VIVO-IN VITRO RAT PERIPHERAL LYMPHOCYTE SISTER CHROMATID 

EXCHANGE ASSAY 

study Number: AA40PA. 130.BTL 

Study Director: Ramadevi Gudi, Ph.D. 

This study has been divided into a series of in-process phases. Using a random 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 (2 1 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 of Good Laboratory Practice and to assure 

that the study is conducted according to the protocol and relevant Standard Operating Procedures. 

The following are the inspection dates, phases inspected, and report dates of QA inspections of this study. 

Inspect On 26-Aug-02 - 26-Aug-02 To 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-03 To Mgmt 24-Jan-03 

Phase Draft Report 

This report describes the methods and procedures used in the study and the reported results accurately reflect 

the raw data of the study. 

DATE 

QUALITY ASSURANCE 

BioReliance Study No. : AA40PA. I30.BTL 3 
PRC - QUALITY ASSURANCE STATEMENT 

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

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

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

as can be reasonably established. 

Reported to 

Study Director and 

Type of Inspection Date(s) of Inspection Management 

GLP Protocol Review 9 Apr 02 9 Apr 02 

Positive Control 21 Aug 02 

Genotoxicity Dose Prep 

& Adm 

Genotoxicity Necropsy 22 Aug 02 

22 Aug 02 

22 Aug 02 

Fran Jannone, B . A., RQAP-GLP 

Quality Assurance Senior Auditor 

Date 

BioReliance Study No.: AA4OPA. 130.BTL 4 
SIGNATURE PAGE 

SCIENTIST 

The following Scientist was responsible for the overall conduct of the SCE portion of this 

study: 

Ramadevi Gudi, Ph.D. 

Principal Investigator 

BioReliance 

Date 

SCIENTIFIC REVIEW 

The following Scientists have 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 

BioReliance Study No. : AA40PA. 130.BTL 
In Vivo-In Vitro Rat Peripheral Lymphocyte Sister Chromatid Exchange Assay 

DRAFT REPORT 

Client: 

Study Director: 

Performing Laboratory 

(Subcontractor): 

BioReliance Principal Investigator: 

Test Substance I.D.: 

Sponsor Project Number: 

BioReliance Study No.: 

Experimental Start Date for SCE: 

Experimental Completion Date for SCE: 

Huntingdon Life Sciences 

Princeton Research Center (PRC) 

Mettlers Road 

East Millstone, New Jersey 08875-2360 

Gary Hoffman, 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 

21 October 2002 

BioReliance Study No. : AA4OPA. 130.BTL 
TABLE OF CONTENTS 

Page 

SUMMARY .......................................................................................................................................................... 8 

PURPOSE ............................................................................................................................................................ 9 

CHARACTERIZATION OF TEST AND CONTROL SUBSTANCES ..................................................... 9 

MATERIALS AND METHODS ........................................................................................................................ 9 

RESULTS AND DISCUSSION ........................................................................................................................ 11 

CONCLUSION .................................................................................................................................................. 12 

REFERENCES .................................................................................................................................................. 13 

DATA TABLES ................................................................................................................................................. 14 

APPENDIX A: CONTRACTING SPONSOR'S EXPOSURE DATA AND ANIMAL DATA ................... 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 following inhalation exposure of rats. 

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 exposed by inhalation to negative (air) control, 2,000, 10,000 or 

20,000 mg/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 mglkg of cyclophosphamide (CP, positive control) by intraperitoneal injection 24 hours 

prior to blood collection. BioReliance personnel collected rat blood samples at 24 hours after 

the end of a 4 week (5 days per week) exposure period. Rat blood samples were cultured and 

processed for the SCE assay at the BioReliance facility. 

A minimum of 25 second division metaphases per animal were scored for SCEs. At least 

100 consecutive metaphases per 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 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 on the 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 

Condensate to induce SCEs in rat peripheral lymphocytes following inhalation exposures. 

CHARACTERIZATION OF TEST AND CONTROL SUBSTANCES 

Information regarding test substance and negative control and their characterizations will 

be provided by Huntingdon Life Sciences. 

Cyclophosphamide (CP, CAS No. 6055-1 9-2, 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 

concentration of 0.5 mg/mL for use as the positive control for the 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 demonstrated by 

acceptable results that met the criteria for a valid test. 

MATERIALS AND 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/m3 of 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 collected into sodium heparin tubes from the abdominal aorta by BioReliance personnel 

on site at Huntingdon Life Sciences. The blood samples were transported to BioReliance on 

the day of collection on wet ice. 

Cell Culture and CoIIection of 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, 100 dmL 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/d of colcemid was added to each flask and incubated for 

approximately 4 hours. Approximately 72 hours after initiation of the 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°C for 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 collected by centrifugation, the supernatant aspirated, and the cells were 

fixed with two washes with approximately 3-5 mL of fixative and stored in fixative overnight 

or longer at approximately 24°C. 

Slide Preparation 

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 

study number, animal number, replicate tube designation and date prepared. 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 fresh by diluting 40 mL of Hoechst 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 

left to air dry. 

Evaluation of SCEs 

Slides were 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 

of toxicity (cell cycle delay). At least 1000 cells were scored for mitotic index per animal. 

BioReliance Study No. : AA40PA. 130.BTL 
Evaluation of Test Results 

The number of SCEs per second-division metaphase cell were presented for each group 

along with the range of SCEsImetaphase for animals within a group. SCE frequencies as 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, cells x 2)+(number M, cells x 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 

evaluation of this 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-level responsive 

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 of the response and the number of exposure levels affected. 

Criteria for Determination of a Valid Test 

The mean SCEs/cell/animal for the positive control animals must be statistically 

increased relative to 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 of the final report, all of the above will be shipped to Huntingdon Life Sciences 

to be archived. All study materials returned to Huntingdon Life Sciences will 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 

evaluation at BioReliance until the final report is issued in accordance with the relevant Good 

Laboratory Practice Regulations. 

RESULTS AND DISCUSSION 

Sister chromatid exchanges involve a symmetrical exchange 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 exposure of rats. 

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 exposed by inhalation to negative (air) control, 2,000, 10,000 or 20,000 mg/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 

days per week) exposure period. Rat blood samples were cultured and processed for the SCE 

assay at BioReliance. 

A minimum of 25 second division metaphases per animal were scored for SCEs. At least 

100 consecutive metaphases per 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 

Generation Time (AGT) and the mitotic index per group for males are presented 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 relative to the negative control group. 

CONCLUSION 

The positive control fulfilled the requirements 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 in rat peripheral 

was concluded to be negative for 

lymphocytes. 

BioReliance Study No. : AA4OPA. 130.BTL 
REFERENCES 

Evans, H.J. (1976) Cytological methods for detecting chemical mutagens, in: A. Hollaender 

(Ed.), Chemical Mutagens, Principles and Methods for their Detection, vol. 4. Plenum Press, 

New York, NY. 

MINITAB User's guide 2: data Analysis and Quality Tools, Release 12 for Windowsm 95 

WindowsNTm February 1998 Chapter 3: Analysis of Variance, 3-1 to3-7. 

Peny P. and Wolff S. (1 974a) New Giemsa method for the differential staining of sister 

chromatid, Nature (London) 25 1, 1 56- 1 5 8 

Wolff S. and Peny P. (1974) Differential Giemsa staining of sister chromatid and the study 

of sister chromatid exchanges without autoradiography. Chromosoma 48,341-353 

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 108 1- 1085 are males and 1591- 1595 are females. 

Number of MI, M2 and M3 cells are obtained from 100 metaphases. 

Mitotic index; number mitotic figures x 100/1000 cells countedkmimal. 

BioReliance Study No. : AAIOPA. 130.BTL 14 
TABLE 2 

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

Gasoline TBA Vapor Condensate 

Male and Female Rats 

1 Number of SCE's Counted I 

I I Animal Number 1 

' Animal numbers 207 1-2075 are males and 258 1-2585 are females. 

Number of M1, M2 and M3 cells are obtained fiom 100 metaphases. 

Mitotic index; number mitotic figures x 100/1000 cells counted/animal. 

BioReliance Study No. : AA40PA. 130.BTL 
TABLE 3 

Number of Sister Chromatid Exchanges per cell at 10,000 rng/m3 of 

Gasoline TBA Vapor Condensate 

Male and Female Rats 

' Animal numbers 3071-3075 are males and 3581-3585 are females. 

Number of MI, M2 and M3 cells are obtained from 100 metaphases. 

Mitotic index; number mitotic figures x 100/1000 cells countedlanirnal. 

BioReliance Study No. : 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 males and 4591-4595 are females. 

Number of MI, M2 and M3 cells are obtained from 100 metaphases. 

Mitotic index; number mitotic figures x 100/1000 cells countedlanimal. 

BioReliance Study No. : 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 Animal Number 

Cell No. 5051 5052 5053 

25 20 28 

20 21 20 

21 23 25 

4 22 24 26 

' Animal numbers 5051-5055 are ma11 ; and 5561-5565 are females. 

Number of M1, M2 and M3 cells are obtained from 100 metaphases. 

Mitotic index; number mitotic figures x 100/1000 cells counterYanimal. 

BioReliance Study No. : AA40PA. 130.BTL 18 
TABLE 6 

Summary of Sister Chromatid Exchange Data -Gasoline TBA Vapor Condensate Male Rats 

Sister Chromatid Exchange data 

Total Range of 

# of # of SCEs Mean Group Group 

Treatment Animal Metaphases SCEs per 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 

Gasoline TBA Vapor Condensate 

10,000 mg/m3 

3071 25 

3072 25 

3073 25 

3074 25 

3075 25 

Gasoline TBA Vapor Condensate 

20,000 mg/m3 

408 1 25 

4082 25 

4083 25 

4084 25 

4085 25 

Cyclophospharnide 

5 mgfkg 

505 1 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 generation time: Number of hours in BrdU X 1001 (number M, cells x I)+(number M, cells x 2)+(number M3 cells x 3) 

BioReliance Study No. : AA40PA. 130.BTL 19 
TABLE 7 

Summary of Sister Chromatid Exchange Data - Gasoline TBA Vapor Condensate Female Rats 

Sister Chromatid Exchange data 

Total Range of 

# of # of SCEs Mean Group Group 

Treatment Animal Metaphases SCEs per 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 

10,000 mg/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 

5 mgk 

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 of all 125 metaphases scored. 

1 = Dunnett's t test (p 4.05) 

Mitotic Index: (Number of cells in mitosis out of 1000 cells) 

AGT=Average generation time: Number of hours in BrdU X 100/ (number M, cells x l)+(number M, cells x 2)+(number M, cells x 3) 

BioReliance Study No. : AA40PA. 130.BTL 20 
Appendix A: Contracting Sponsor's Exposure Data and Animal Data 

BioReliance Study No. : AA40PA. 130.BTL 2 1 
Huntingdon Life Sciences 00-6 13 1 Page 1 

21 1-TBA-S Draft Final Report 

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

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

the procedures used for test atmosphere generationlcharacterization and animal 

evaluations. 

Animal Exposure and Animal Data 

Preface 

STUDY DATES: Date of Animal Receipt: 15 July 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. Summary of Clinical Observations (pretest only) ............................................................ C. Mean Body Weights (grams) ........................................................................................... 

D. Mean Body Weight Change from Baseline (grams) ........................................................ E. Mean Feed Consumption Values (grams/kg/day) ............................................................ F. Individual Clinical Observations (pretest only) ............................................................... G. Individual Body Weights (grams) .................................................................................... H. Individual Body Weight Change from Baseline (grams) ................................................. 

I. Individual Feed Consumption Values (gramskglday) ..................................................... 

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

Chamber Monitoring Results 

Cumulative Exposure Record 
Appendix A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY STUDY IN RATS 00-6131 

Chamber Monitoring Results 

Date Exposure 

Number 

1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Mean 

S.D. 

Cumulative Exposure Record 

Group 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 

ns 

TMC 

Owm3) 

7.27E-03 

5.51 E-03 

2.1 2E-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 

2.8 
Appendix A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-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 

21 50 

2110 

2080 

1950 

2020 

21 60 

2020 

2020 

21 90 

2090 

2090 

21 40 

2200 

2070 

21 30 

alytical Chamber Concentration 

Individual 

ms 

TMC 

(mglm") 

4.69E-03 

9.38E-03 

2.35E-02 

Humidity 

("4 

50 

50 

48 

49 

5 1 

5 1 

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 

21 35 

21 88 

1953 

1845 

2043 

2023 

201 3 

2070 

2090 

2068 

21 83 

MMAD 

Olm) 

2.312 

4.144 

0.7525 

eterrnina 

GSD 

Mean 
Appendix A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-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 

21 05 

2060 

1935 

1830 

2048 

21 23 

2020 

2018 

1970 

2203 

1968 

2088 

21 65 

2008 

1995 

2028 

2073 

2023 

2025 

alytlcal Chamber Concentration 

Individual 

I 

MMAD 

Olm) 

3.582 

2.398 

0.7703 

Particle Size 

!terminal 

GSD 

ns 

TMC 

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 

3.4 
Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY 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 

981 8 

10210 

lo580 

10200 

10010 

9763 

10190 

10220 

9463 

10180 

10020 

10440 

9868 

erminati 

MMAD GSD 

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 GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-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 - 10,000 mg/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 

881 0 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 GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY 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 

1.81 E-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 

5 1 

54 

50.6 

3.0 
Table A GASOLINE TBA VAPOR CONDENSATE: A 13-WEEK WHOLE-BODY INHALATION TOXICITY 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 

Cumulative Exposure 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 Mean 
Huntingdon 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 LIMITS 
Huntingdon 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 LIMITS 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE C 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

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 differences 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE C 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

FEMALES MEAN BODY WEIGHTS (GRAMS) 

DOSE GROUP: I I I I11 I V 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 differences 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE D 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MALES MEAN BODY WEIGHT CHANGE FROM BASELINE (GRAMS) ................................................................................................................................... 

DOSE GROUP: 

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 differences 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE D 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

DAY -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 differences 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE E 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

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

DOSE GROUP: I 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 differences 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE E 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

FEMALES 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 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 differences 
Huntingdon 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 1 
Huntingdon 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 2 
Huntingdon 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-PRESENT 
Huntingdon 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-PRESENT 
Huntingdon 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-PRESENT 
Huntingdon 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-PRESENT 
Huntingdon 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-PRESENT 
Huntingdon 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-PRESENT 
Huntingdon 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 9 
Huntingdon 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 10 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE G 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

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. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE G 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 3 

TABLE G 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

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

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 4 

TABLE G 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

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

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. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 5 

TABLE G 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

MALES GROUP V SCE+CONTROL ------------------------------------------------------------------.----.--------

DAY OF STUDY 

ANIMAL# -3 4 11 18 25 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 6 

TABLE G 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

FEMALES GROUP I 0 MG/M3 

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

DAY OF STUDY 

ANIMAL# - 3 4 11 18 25 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 7 

TABLE G 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

FEMALES GROUP I1 2000 MG/M3 

DAY OF STUDY 

ANIMAL# - 3 4 11 18 25 

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. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 8 

TABLE G 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMLTNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 9 

TABLE G 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHTS (GRAMS) 

FEMALES GROUP IV 20000 MG/M~ ---------------------------------------------------------------------------.----

DAY OF STUDY 

ANIMAL# - 3 4 11 18 2 5 

MEAN 

S.D. 

N 
Huntingdon 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. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE H 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND 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. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE H 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

MALES GROUP I1 2000 MG/M3 ................................................................................................................................... 

DAY OF STUDY 

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

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 3 

TABLE H 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

MALES GROUP I11 10000 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 4 

TABLE H 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

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

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 5 

TABLE H 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND (-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

MALES GROUP V SCE+CONTROL 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 6 

TABLE H 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

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. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 7 

TABLE H 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

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

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 8 

TABLE H 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL BODY WEIGHT CHANGE FROM BASELINE (GRAMS) 

FEMALES GROUP I11 10000 MG/M3 

DAY OF STUDY 

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

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 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. 

N 
Huntingdon 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 10 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 1 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

MALES GROUP I 0 MG/M3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 3 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

MALES GROUP I11 10000 MG/M3 

MEAN 

S.D. 

N 

SF=Spilled Feeder 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 4 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 128 107 9 3 83 

S.D. 8.1 4.4 1.5 1.8 

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

Genotoxicity Sub-Group 

PAGE 5 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

MALES GROUP V SCE+CONTROL 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 6 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 7 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 8 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

FEMALES GROUP I11 10000 MG/M3 

MEAN 

S.D. 

N 

OF STUDY 

11 18 --------------- 

105 97 

106 98 

105 110 

14 5 SF 

108 103 

SF=Spilled Feeder 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 9 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

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

DAY 

ANIMAL# 4 ....................... 

4591 119 

4592 119 

4593 124 

4594 123 

4595 127 

OF STUDY 

11 18 2 5 

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

104 9 2 8 8 

101 9 4 89 

107 101 93 

103 101 9 5 

111 104 9 3 

MEAN 

S.D. 

N 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 10 

TABLE I 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

INDIVIDUAL FEED CONSUMPTION VALUES (GRAMS/KG/DAY) 

FEMALES GROUP V SCE+CONTROL ..................................................................................................................... 

MEAN 

S.D. 

N 

DAY OF STUDY 

4 11 18 

SF=Spilled Feeder 
Huntingdon 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 2 7 

1083 TERMINAL SACRIFICE 22-AUG-02 3 2 7 

1084 TERMINAL SACRIFICE 22-AUG-02 3 2 7 

1085 TERMINAL SACRIFICE 22-AUG-02 3 27 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 2 

TABLE J 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

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-02 
Huntingdon Life Sciences 00-6131G 

Genotoxicity Sub-Group 

PAGE 3 

TABLE J 

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

TOXICITY STUDY IN RATS WITH NEUROTOXICITY ASSESSMENTS AND 4-WEEK 

IN VIVO GENOTOXICITY AND IMMUNOTOXICITY ASSESSMENTS 

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 2 7 

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 SACRIFICE 
Huntingdon 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 ASSESSMENTS 
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 

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

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 ASSESSMENTS 
Huntingdon 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 2 7 

5565 TERMINAL SACRIFICE 22-AUG-02 3 27 ------------.---------------------------------------------------------.--------------