# STUDY NO. 00

- 6130

SPONSOR STUDY NO. 211

- DIPE -S

GASOLINE DIPE VAPOR CONDENSATE:

A 13

- WEEK WHOLE

- BODY INHALATION TOXICITY STUDY IN RATS

WITH NEUROTOXICITY A SSESSMENTS AND 4

- WEEK IN VIVO

GENOTOXICITY AND IMMUNOTOXICITY ASSESSME NTS

Final Report

Volume I of IV

Submitted to: American Petroleum Institute (API)

1220 L Street, Northwest

Washington, DC 20005

- 4070

Attn: Thomas M. Gray, M.S., D.A.B.T.

Date: 31 May 2013

Page 1 of 1446

# Huntingdon Life Sciences 00-6130 Page 5

211- DIPE -S Final Report

GASOLINE DIPE VAPOR CONDENSATE:

A 13

- WEEK WHOLE

- BODY INHALATION TOXICITY STUDY IN RATS

WITH NEUROTOXICITY ASSESSMENTS AND 4

- WEEK IN VIVO

GENOTOXICITY AND IMMUNOTOXICITY ASSESSME NTS

SUMMARY

This study was designed to assess the potential inhalation toxicity of Gasoline DIPE

Vapor Condensate when administered via whole

- body exposures to Sprague Dawley rats

for 13 weeks followed by a 4 week recovery period. The assessments included routine

tox icology parameters as well as detailed evaluations of neurotoxicity parameters

(presented in this report except GFAP evaluations appended to this report) and

genotoxicity and immunotoxicity parameters (appended to this report). The test

substance was admi nistered (using a nitrogen enriched air mixture) at target

concentrations of 2000, 10000 and 20000 mg/m 3 

# for 6 hours/day, generally 5 days/week

for 13 weeks (at least 65 exposures). In addition, an Air Control group received nitrogen

enriched air only while in chamber. Exposure levels were determined using an infra -red

spectrophotometer 4 times per chamber per day. Additionally, the test substance’s major

components were assayed once per chamber per week. Particle size distribution

measurements were als o made once per chamber per week using a TSI Aerodynamic

Particle Sizer.

Viability checks were performed twice daily to check for mortality and signs of severe

toxic or pharmacologic effects. Physical observations and body weights were collected

twice p retest and weekly during the study period. Feed consumption measurements were

obtained beginning the week prior to treatment initiation and weekly during the study.

Ophthalmoscopic examinations were conducted pretest and at study termination.

Functional observational batteries and motor activity assessments were performed pretest

and after 4, 8 and 13 weeks of exposure. Hematology, coagulation and clinical chemistry

studies were performed at 4 weeks and termination of the exposures. After 13 weeks of

e xposures, all animals were sacrificed except recovery animals, which were sacrificed

after an additional 4

- week recovery period. Selected organs were weighed and

organ/body weight and organ/brain weight ratios calculated. Complete macroscopic

postmortem examinations were performed on all animals. Histopathological evaluations

(including neuropathology evaluations) of all selected or prescribed tissues were

conducted on all Air Control and 20000 mg/m 3 

# exposed animals at the Terminal interval

but no evalua tions were made at the Recovery interval. Lungs and gross lesions were

examined for all animals at the Terminal interval.

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SUMMARY

The mean ( ± standard deviation) analytical exposure concentrations of Gasoline DIPE

Vapor Condensate were determined to be 0 ± 0, 1995 ± 132, 10030 ± 692 and 20120

± 869 mg/m 3 

# for the Air Control and the exposure groups, respectively. The analytically

measured exposure levels of the airborne test substance were accept ably close, in the

opinion of the study director, to the targeted exposure levels. Chamber environmental

conditions averaged 24 ° C temperature and 50% relative humidity. Particle sizing results

indicated that the atmospheres were essentially vapor only, as expected, since there was

no substantial difference between the particulate levels in the test substance chambers

and the Air Control chambers. Analysis of the major components in the neat test

substance and the test atmospheres showed a reasonably close comparison between the

neat test substance and the vaporized test substance. This data demonstrated that the test

animals wer e exposed, as expected, to all of the major components of the test substance

in their reasonably proper proportion. The data was consistent from week

- to

- week

during the study indicating stability of the test substance and the atmosphere generation

techniq ues.

There was no test substance

- related mortality throughout the exposure and recovery

phases of the study. The test animals were unremarkable during the in

- chamber exposure

periods. During the non

- exposure periods, an exposure

- related increase in the frequency

of red nasal discharge observations was seen in the 20000 mg/m 3 

# exposed animals

compared to the Air Control animals and the other test substance exposed animals

starting as early as Week 2 for the males and Week 4 for the females. These observat ions

abated during the recovery period. There were no indications of treatment

- related ocular

disease in the test substance exposed animals. There were no exposure

- related

differences in absolute body weights or in body weight changes or in feed consumpt ion

in the test substance exposed animals compared to the Air Control animals. Test

substance exposure was not associated with any change in counts of the 26 nominal FOB

measures, or the 4 continuous FOB measures or with changes in motor activity.

Ther e were exposure

- related differences in hematology values in test substance exposed

animals compared to the Air Control animals at the 4 th 

# week and terminal and recovery

intervals. These differences were limited to statistically significant decreases (21-2 3%)

in reticulocyte counts for the 20000 mg/m 3 

# exposed male animals which occurred at both

exposures period intervals (4 th 

# week and terminal) and at termination of the recovery

interval. Statistical ly significant (13- 17%) decreases also occurred in reticu locyte counts

for the 2000 and 10000 mg/m 3 

# exposed male animals but only at the 4 th 

# week interval.

Similar differences did not occur in female animals at any exposure level and any interval

but there were s tatistical ly significant (35

- 36%) decreases in ab solute monocytes for the

20000 mg/m 3 

# exposed fe male animals but only at the 4 th 

# week and recovery intervals

. There were no exposure

- related differences in coagulation values in test substance

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211- DIPE -S Final Report

SUMMARY

exposed animals compared to the Air Control animals at the 4th week and Terminal

intervals. There were exposure

- related differences in clinical chemistry values in the test

substance exposed animals compared to the Air Control animals at the 4 th 

# week and

terminal and recovery intervals. Decreases, relative to control values, occurred in several

serum enzymes for test substance

- exposed groups after 4 or 13 weeks of exposure and/or

at the end of the recovery period. Enzymes affected were aspartate aminotransferase,

lactate dehydrogenase, creatine kinase and sorbitol d ehydrogenase. While some of the

differences (e.g. lactate dehydrogenase and sorbitol dehydrogenase) were consistently

statistically significant at the 4 and 13 weeks and/or recovery intervals, decreases in these

enzymes are not usually considered an adverse response to treatment and there were no

corresponding histopathology findings.

There were no exposure

- related differences in organ weights and brain measurements in

the test substance exposed animals compared to the Air Control animals at the Terminal

and/or Recovery intervals. No gross abnormalities related to test substance exposure

were evident at necropsy examination. Microscopic findings that were considered

exposure- related were found only in the kidneys of male animals exposed to

20000 mg/m 3 

# of Gasoline DIPE Vapor Condensate following the Terminal sacrifice.

These renal histopathologic changes were consistent with hyaline droplet nephropathy,

attributable to accumulation of alpha

- 2 microglobulin within renal tubular epithelial cells.

This spec ies

- and gender

- specific change has been well documented in male rats exposed

to a variety of hydrocarbon compounds and is not considered relevant to humans. No test

substance related histopathologic changes were noted in other protocol

- specified tissues,

including lung, nasoturbinates and larynx. No microscopic changes attributable to test

substance exposure were observed in brain, spinal cord, eyes with optic nerve, peripheral

nerves or ganglia among the 20000 mg/m 3 

# exposed satellite animals.

In conclusion, thirteen weeks of exposure of rats to Gasoline DIPE Vapor Condensate

resulted in hydrocarbon nephropathy in male animals exposed to 20000 mg/m 3 

# of vapor.

However, this finding has been generally accepted (US EPA 1991. Alpha 2

microglobulin: association with chemically induced renal toxicity and neoplasia in male

rat. In Risk Assessment Forum, p.85. US Govt. Printing Office, Washington, DC.) not to

be relevant to human risk assessment. Thirteen weeks of exposure of rats to Gasoline

DIPE Vapor Conden sate also resulted in increased red nasal discharge and slight, non

reversible decreases in reticulocyte values (at the 4 th 

# week and terminal and recovery

intervals) in the male animals exposed to 20000 mg/m 3 

# of vapor. Therefore, the

10000 mg/m 3 

# exposure level (excluding male rat nephropathy) was considered a no

observable adverse effect level (NOAEL). The NOAEL for other satellite study

endpoints are included in the Table on page 8.

# Huntingdon Life Sciences 00-6130 Page 8

211- DIPE -S Final Report

SUMMARY

Study Type

[Laboratory] Duration of

Exposure Basis for NOAEL NOAEL

Inhalation Subchronic 1



# [HLS,

East Millstone, NJ] 13 weeks Positive:

altered increases in red nasal

discharge and slight, nonreversible decreases in

reticulocyte values in the male

animals exposed to

20000 mg/m 3 

# 10000mg/m 3



# Neurobehavioral and

Neuropathol ogic Effects 4 weeks Negative 20000mg/m 3



# Micronucleus 2



# [HLS,

Eye Suffolk, England] 4 weeks Negative 20000mg/m 3



# Sister Chromatid

Exchange 3



# [BioReliance,

Rockville, MD] 4 weeks Negative 20000mg/m 3



# Immunotoxicity 4



# [ImmunoTox Inc,

Richmond, VA] 4 week s Positive:

significant decrease in the IgM

antibody- forming cell (AFC)

response to the T-dependent

antigen, sheep erythrocytes, in

the female animals exposed to

20000 mg/m 3 

# 10 000mg/m 3



# GFAP 5



# [Molecular

Neurotoxicology Lab -

NIOSH,

Morgantown, VA] 13 weeks Negative 20000mg/m 3

1 

# Light Hydrocarbon Nephropathy observed in males at 20000mg/m 3 

# exposure level

2 

# Appendix X: Micronucleus Satellite Study Report

3 

# Appendix Y: Sister Chromatid Exchange Satellite Study Report

4 

# Appendix Z: Immunotoxicity Satellite Study Report

5 

# Appendix AA: GFAP Satellite Study Report

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211- DIPE -S Final Report

TABLE OF CONTENTS

VOLUME I

COVER PAGE .....................................................................................................................1

STATEMENT OF COMPLIANCE .....................................................................................2

SIGNATURE PAGE ............................................................................................................3

QUALITY ASSURANCE STATEMENT ..........................................................................4

SUMMARY .........................................................................................................................5

TABLE OF CONTENTS .....................................................................................................9

1. 

# INTRODUCTION ................................................................................................13

2. 

# MATERIALS AND METHODS

2.1. Study Management ....................................................................................13

2.2. Study Dates ................................................................................................13

2.3. Experimental Outline .................................................................................15

2.4. Test Substance ...........................................................................................17

2.5. Test Animals ..............................................................................................18

2.6. Animal Assignment ...................................................................................19

2.7. Animal Identification .................................................................................20

2.8. Veterinary Care ..........................................................................................20

2.9. Husbandry During Non-Exposure Periods.................................................20

2.10. Husb andry During Exposure Periods .........................................................22

2.11. Test Substance Administration and Chamber Operation ...........................23

2.12. Experimental Evaluations ..........................................................................27

2.13. Neurobehavioral Studies ............................................................................29

2.14. Clinical Pathology ......................................................................................31

2.15. Genotoxicity Evaluations ...........................................................................35

2.16. Immunotoxicity Evaluations ......................................................................36

2.17. Postmortem Evaluations ............................................................................37

2.18. Neuropathology Sacrifice ...........................................................................41

2.19. GFAP Evaluations .....................................................................................42

2.20. Statistical Analysis .....................................................................................43

2.21. Data Storage ...............................................................................................45

2.22. Regulatory References ...............................................................................45

2.23. Protocol Deviations ....................................................................................47

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TABLE OF CONTENTS

3. 

# RESULTS AND D ISCUSSION

3.1. Chamber Monitoring ..................................................................................48

3.2. Mortality .....................................................................................................50

3.3. Clinical Observations .................................................................................50

3.4. Ophthalmoscopic Examinations ................................................................51

3.5. Body Weights .............................................................................................51

3.6. Feed Consumption .....................................................................................51

3.7. Neurobehavioral Studies ............................................................................51

3.8. Clinical Pathology ......................................................................................53

3.9. Organ W eights and Brain Measurements ..................................................54

3.10. Pathology ...................................................................................................55

4. 

# CONCLUSION ....................................................................................................57

REFERENCES ..................................................................................................................58

FIGURES

1. Chamber Generation System and Whole-Body Exposure Chamber ...........................61

2. Mean Body Weights – Males (Main/Recovery) ..........................................................63

3. Mean Body Weights – Females (Main/Recovery) .......................................................64

4. Mean Feed Consumption – Males (Main/Recovery) ...................................................65

5. Mean Feed Consumption – Females (Main/Recovery) ...............................................66

GENERAL PREFACE ......................................................................................................67

TABLES

1. Summary of In -Chamber Observations .......................................................................68

2. Summary of Clinical Observations ..............................................................................73

3. Summary of Ophthalmoscopic Findings ......................................................................82

Ophthalmologist Reports ..............................................................................................83

Summary of Ophthalmology Observations ..................................................................87

4. Mean Body Weights .....................................................................................................95

5. Mean Body Weight Change from Baseline ................................................................105

6. Mean Feed Consumption Values ...............................................................................117

7. Summary of FOB and Motor Activity Analyses ........................................................127

8. Mean Motor Activity Values .....................................................................................139

9. Summary of Functional Observational Battery Evaluations ......................................147

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TABLE OF CONTENTS

10. Mean Hematology Values ..........................................................................................179

11. Mean Coagulation Values ..........................................................................................192

12. Mean Clinical Chemistry Values ...............................................................................197

13. Mean Organ Weights and Brain Measurements ........................................................216

14. Incidence Summary Report for Gross Necropsy Observations ..................................243

15. Incidence Summary of Microscopic Findings with Severity Levels ..........................247

APPENDICES

A. Chamber Monitoring Results .....................................................................................267

B. Individual Clinical Observations ...............................................................................292

C. Individual Ophthalmology Observations ...................................................................321

D. Individual Body Weights ...........................................................................................361

Volume II

COVER PAGE .................................................................................................................381

TABLE OF CONTENTS .................................................................................................382

APPENDICES

E. Individual Body Weight Change from Baseline. .......................................................383

F. Individual Feed Consumption Values ........................................................................399

G. Individual Motor Activity Values ..............................................................................419

H. Individual Functional Observational Battery Evaluations .........................................435

I. Individual Hematology Values ..................................................................................472

J. Individual Coagulation Values ...................................................................................513

K. Individual Clinical Chemistry Values ........................................................................530

L. Individual Organ Weights and Brain Measurements .................................................591

M. Individual Animal Gross and Microscopic Observations ..........................................630

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TABLE OF CONTENTS

Volume III

COVER PAGE .................................................................................................................816

TABLE OF CONTENTS .................................................................................................817

APPENDICES

N. Equipment List ...........................................................................................................818

O. Miran Calibration .......................................................................................................821

P. Chamber Distribution Records ..................................................................................824

Q. Testing Room and Chambers Environmental Monitoring .........................................826

R. Animal Terminati on History ......................................................................................828

S. Analytical Report .......................................................................................................844

T. Neurobehavioral Positive Control ..............................................................................869

U. Neuropathology Historical Control ............................................................................962

V. Protocol and Protocol Amendment ............................................................................968

W. Testing Facility Personnel ........................................................................................1056

Volume IV

COVER PAGE ...............................................................................................................1057

TABLE OF CONTENTS ...............................................................................................1058

APPENDICES

X. Micronucleus Satellite Study Report ......................................................................1059

Y. Sister Chromatid Exchange Satellite Study Report ................................................1159

Z. Immunotoxicity Satellite Study Report ..................................................................1251

AA. GFAP Satellite Study Report ..................................................................................1315

BB. Report Amendments ...............................................................................................1446

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1. INTRODUCTION

This study was designed to assess the potential inhalation toxicity of Gasoline

DIPE Vapor Condensate when administered via whole

- body exposures to rats for

at least 13 weeks followed by a 4 week recovery period. The assessmentincluded

routi ne toxicology parameters as well as detailed evaluations of neurotoxicity,

genotoxicity and immunotoxicity parameters.

2. MATERIALS AND METHOD S

2.1. STUDY MANAGEMENT

2.1.1. SPONSOR

American Petroleum Institute (API)

1220 L Street, Northwest

Washington, D.C. 20005-4070

2.1.2. SP ONSOR REPRESENTATIVE

Thomas M. Gray, M.S., D.A.B.T.

2.1.3. TESTING FACILITY

Huntingdon Life Sciences (HLS)

Mettlers Road

East Millstone, New Jersey 08875 -2360

2.1.4. STUDY DIRECTOR

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

2.1.5. CONTRIBUTORS

Departmental supervisory personnel are liste d on the personnel

page of this report (Appendix W ).

2.2. STUDY DATES

2.2.1. STUDY INITIATION

17 December 2001 (Date Study Director signed the Protocol)

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211-DIPE-S Final Report

2.2.2. DATE OF ANIMAL RECEIPT

24 January 2002 (main/recovery/neuropathology study animals)

2.2.3. EXPOSURE INITIATION

12 February 2002 (Experimental Start Date)

2.2.4. EXPOSURE TERMINATION

13 - 14 May 2002

2.2.5. TERMINAL SACRIFICE

13 - 14 May 2002 (main study animals)

16 - 17 May 2002 (neuropathology study animals)

2.2.6. RECOVERY SACRIFICE

14 June 2002

2.2.7. EXPERIMENTAL TERMINATION

29 May 2013 (Date of last data collection = Date final report is

signed by the Pathologist)

2.2.8. STUDY COMPLETION

31 May 2013 (Date Final Report is signed by the Study Director)Huntingdon Life Sciences00-6130Page 15211-DIPE-S FinalReport2.3.EXPERIMENTAL OUTLINEGroupExposureLevelaNumber of Animals (MAIN and RECOVERY STUDIES)(mg/m3)InitialbClinical Laboratory StudiesNecropsyMicroscopic Pathology4 Weeks &TerminalRecoverycTerminal RecoverycTerminal RecoverycM F M F M F M F M F M F M FI (control) 0 (air only) 20 20 10 10 10 10 10 10 10 10 10 10 10 10II (low) 2,000 10 10 9d10 - - 9d10 - - 9d10 - -III (mid) 10,000 10 10 1010 - - 1010 - - 1010 - -IV (high) 20,000 20 20 10 10 10 10 10 10 1010 10 10 1010aExposures were 6 hours/day, generally 5 days/week for 13 weeks for at least 65 exposures for Main and Recovery Studies. Animals were notexposed on test days when FOB and motor activity evaluations were performed.bThis number of animals includes 10/sex/group designated for the Terminal Sacrifice (Main Study animals) and 10/sex/group (Groups I & IV only)designated for Recovery after the Terminal Sacrifice (Recovery Study animals).cThe recovery period was 4 weeks. It wasdecided by the Sponsor that Clinical Laboratory Studies would not be performed on recovery group animalsif no significant effects were observed in animals during the exposure period. Past testing on gasoline, gasoline blending streams, and neatoxygenates had not demonstrated delayed expression of toxicity after exposure was terminated that had not been seen in exposed animals. For thisstudy, only hematology and clinical chemistrywere performed for the Recovery interval.dA Group II male animal (#2070) was euthanized on 29April02 as a result of an accidental injury. Since this animal was 1 of 5 male animalsdesignated for GFAP evaluation after 13 weeks of exposure, it wasdecided to replace this animal with 1 of 10 male animals designated for MainStudy evaluations.M = Male; F = Female; The first day of exposure was Day 0.Huntingdon Life Sciences00-6130Page 16211-DIPE-S FinalReportEXPERIMENTAL OUTLINE (CONTINUED)GroupExposureLevelaNumber of Animals (SATELLITE STUDIES)(mg/m3)InitialbNeuropathologyGFAPImmunotoxicityGenotoxicity(Micronucleus and SCE)M F M F M F M F M FI (control) 0 (air only) 15 25 5 5 5 5 - 10 5 5II (low) 2,000 15 25 5 5 5 5 - 10 5 5III (mid) 10,000 15 25 5 5 5 5 - 10 5 5IV (high) 20,000 15 25 5 5 5 5 - 10 5 5aExposures were 6 hours/day, generally 5 days/week for 13 weeks for at least 65 exposures for Neuropathology and GFAP Studies. Exposures were 6hours/day, generally 5 days/week for 4 weeks for 20 exposures for Genotoxicity and Immunotoxicity Studies. Animals were not exposed on testdays when FOB and motor activity evaluations were performed.bThis number of animals included 5/sex/groupdesignated for neuropathology after the Terminal Sacrifice, 5/sex/group designatedfor GFAPevaluation after the Terminal Sacrifice, 10 females/group designated for Immunotoxicity (PFC) evaluations following a 4 week exposure period and5/sex/group designated for Genotoxicity (Micronucleus and Sister Chromatid Exchange) evaluations following a 4 week exposure period. For theImmunotoxicity and Genotoxicity evaluations,separate groups of animals (10 females and 5/sex/evaluation, respectively) were used as positivecontrol animals for these evaluations. For Immunotoxicity evaluations, positive control rats weregiven a dose of 50 mg/kg Cyclophosphamide (CP)intraperitoneally once per day for 4 days prior to sacrifice.For Genotoxicity evaluations, positive control rats were given adose of 5 mg/kg (SCE)and 40 mg/kg (Micronucleus) CP intraperitoneally within 24 hours prior to sacrifice. These positive control animals were not chamber exposed priorto dosing and sacrifice.M = Male; F = Female; The first day of exposure was Day 0.GFAP = Glial Fibrillary Acidic Protein; SCE = Sister Chromatid Exchange; PFC = Plaque Forming Cell

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2.4. TEST SUBSTANCE

Gasoline DIPE Vapor Condensate

2.4.1. SUPPLIER

Chevron Global Technology Services Co.

100 Chevron Way

Richmond, CA 94802-9627

2.4.2. LOT NUMBER

API 01-06

2.4.3. PURITY

100% Gasoline DIPE Vapor Condensate

2.4.4. DESCRIPTION

Colorless liquid

2.4.5. DATE RECEIVED

7 December 2001

2.4.6. EXPIRATION DATE

Not available, stable per MSDS. The test substance stability was

tested concurrently with the st udy and the results (see Appendix S)

showed it to be stable for the duration of the study.

2.4.7. ANALYSIS

Documentation of the identity, strength, composition, and method of

synthesis, fabrication and/or derivation of the batch of the test

substance and the maintenance of these records was the

responsibility of the Sponsor.

2.4.8. STORAGE

Room temperature

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2.4.9. DISPENSING

The test substance was received from Chevron Research Company

in 100-gallon cylinders. Since only 5-gallon cylinders were

practical to be used for exposure operations, the test substance was

dispensed, as needed, at the Testing Facility from the 100 gallon

cylinders into 5-gallon cylinders using nitrogen pressurization.

2.4.10. ARCHIVAL SAMPLE

A sample from the lot of test substa nce is stored in the Archives of

the Testing Facility under conditions specified for test substance

storage.

2.4.11. DISPOSITION

The unused portion of the test subs tance as well as any empty test

substance containers were returned to the sponsor on

11 January 07. Empty test substance containers were returned to

the sponsor on an as needed basis. The sponsor was responsible

for tracking their disposition.

2.5. TEST ANIMALS

2.5.1. SPECIES

Albino Rats (Outbred) VAF/Plus 



# Sprague-Dawley – derived [Crl:CD ® 

# (SD) IGS BR]

2.5.2. SUPPLIER

Charles River Laboratories

Kingston, New York 12484

2.5.3. JUSTIFICATION FOR TEST ANIMAL SELECTION

The rat is a rodent animal model commonly utilized in toxicity

studies as recommended in the referenced guidelines. In addition,

a historical database is available for comparative evaluation.

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2.5.4. NUMBER OF ANIMALS

Placed on test:

310 total (130 males, 180 females)

2.5.5. AGE AT RECEIPT AND AT INITIATION OF EXPOSURE

Study Phase Age at Receipt

(weeks) Age at Exposures Initiation

(weeks)

Main/Recovery ~6 ~9

Neuropathology ~6 ~9

GFAP ~6 ~9

Genotoxicity ~4 ~6

Immunotoxicity ~4 ~7

Note: Females were nulliparous and non-pregnant.

2.5.6. WEIGHT AT INITIATION OF EXPOSURES (GRAMS)

Study Phase Male Female

Mean Range Mean Range

Main/Recovery 306 277-351 222 192-263

Neuropathology 301 254-331 226 201-257

GFAP 308 280-336 217 188-240

Genotoxicity 123 108-138 113 95-130

Immunotoxicity - - 182 165-204

Individual weights of animals placed on test were within ± 20% of

the mean weight for each sex.

2.5.7. ACCLIMATION PERIOD

Animals were acclimated for up to three weeks. All animals were

examined during the acclimation period to confirm suitability for

study.

2.6. ANIMAL ASSIGNMENT

More animals than required for the study were purchased and acclimated.

Animals considered suitable for study on the basis of pretest physical

examinations, body weight data and pr etest ophthalmology evaluations were

randomly assigned by a computerized random sort program, by sex, to

# Huntingdon Life Sciences 00-6130 Page 20

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control or treated groups in an attempt to equalize mean group body

weights. The Main/Recovery study and each of the Neuropathology, GFAP,

Genotoxicity and Immunotoxicity studies were randomized separately.

Disposition of all animals not utilized in the study is maintained in the study

file.

2.7. ANIMAL IDENTIFICATION

Each rat was identified with a metal ear tag bearing its assigned animal

number. The assigned animal number plus the study number comprised

the unique animal number for each animal. When a tag was lost, it was

replaced. In addition, each cage was provided with a cage card, which

was color-coded for exposure level identification and contained study

number and animal number information.

2.8. VETERINARY CARE

Animals were monitored by the t echnical staff for any conditions

requiring possible veterinary care.

2.9. HUSBANDRY DURING NON-EXPOSURE PERIODS

2.9.1. FACILITIES MANAGEMENT/ANIMAL HUSBANDRY

Currently acceptable practices of good animal husbandry were

followed e.g., Guide for the Care and Use of Laboratory Animals ;

National Academy Press, 1996. Huntingdon Life Sciences, East

Millstone, New Jersey is fully accredited by the Association for

Assessment and Accreditation of Laboratory Animal Care

International (AAALAC).

2.9.2. HOUSING

Animals younger than 5 weeks at receipt were housed 2/cage during

the initial week of acclimation. Animals 5 weeks or older were

housed individually in suspended stainless steel wire mesh cages.

2.9.3. FEED

Certified Rodent Diet, No. 5002; (Meal) (PMI Nutrition

International, St. Louis, Missouri) was available without

restriction. Fresh feed was presented weekly.

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2.9.4. FEED ANALYSIS

Analysis of each feed lot used during this study was performed by

the manufacturer. Results are maintained on file by the

manufacturer. There were no known contaminants in the feed

which were expected to interfere with the results of this study.

2.9.5. WATER

Water (Elizabethtown Water Company, Westfield, New Jersey)

was available without restriction via an automated watering

system.

2.9.6. WATER ANALYSIS

Monthly water analyses are conducted by Elizabethtown Water

Company, Westfield, New Jersey (Raritan-Millstone Plant) to

ensure that water meets standards specified under the EPA Federal

Safe Drinking Water Act Regula tions (40 CFR Part 141). In

addition, water samples are collected biannually from

representative rooms in the Testing Facility; chemical and

microbiological water analyses are conducted on these samples by

a subcontract laboratory. Results of all water analyses are

maintained on file at the Testing Facility. There were no known

contaminants in the water, which were expected to interfere with

the results of this study.

2.9.7. ENVIRONMENTAL CONDITIONS

Light/Dark Cycle

A twelve hour light/dark cycle controlled via an automatic timer

was provided.

Temperature

Temperature was monitored in accordance with Testing Facility

SOPs and maintained within the specified range to the maximum

extent possible.

Desired: 18 to 26 ° C

Actual: 19 to 25 ° C

Daily Average: 19 to 23 ° C

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Relative Humidity

Relative humidity was monitored in accordance with Testing

Facility SOPs and maintained with in the specified range to the

maximum extent possible. Excurs ions outside the specified ranges

were not considered to have affected the integrity of the study.

Desired: 30 to 70%

Actual: 25 to 76%

Daily Average: 40 to 59%

2.10. HUSBANDRY DURING EXPOSURE PERIODS

2.10.1. HOUSING

Animals were individually housed in stainless steel, wire mesh

cages within a 1000 Liter stainl ess steel and glass whole-body

exposure chamber. Two chambers were used for each exposure

level, in general one for male animals and one for female animals,

with the placement of the animals in the whole-body exposure

chambers rotated weekly to en sure uniform exposure of the

animals. A description of the animal rotation is included in the

raw data.

2.10.2. FEED

None was provided during exposure.

2.10.3. WATER

None was provided during exposure.

2.10.4. ENVIRONMENTAL CONDITIONS

Chamber static pressure and airflow were monitored continuously

and recorded every half-hour during the exposure. Chamber

temperature and relative humidity were also monitored

continuously and recorded every half-hour during the exposure and

maintained, to the maximum extent possible, within the ranges

presented below. Excursions outside the specified ranges were not

considered to have affected the integrity of the study. `

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Temperature

Desired: 20 to 24 ° C

Actual: 21 to 28 ° C

Relative Humidity

Desired: 40 to 60%

Actual: 37 to 69%

2.11. TEST SUBSTANCE ADMINISTRATION AND CHAMBER

OPERATION

2.11.1. ROUTE OF ADMINISTRATION

Inhalation via whole-body exposures.

2.11.2. JUSTIFICATION FOR ROUTE OF ADMINISTRATION

The inhalation route is one of the potential routes of human

exposure to this test substance and is the route specified in the

referenced guidelines.

2.11.3. FREQUENCY OF ADMINISTRATION

Once daily, 6 hours/day.

2.11.4. DURATION OF ADMINISTRATION

The test substance was administered for 6 hours/day, generally

5 days per week for 13 weeks for at least 65 exposures for

Main/Recovery/Neuropathology/GFAP Studies. Animals were not

exposed on test days when FOB and motor activity evaluations

were performed. Animals for the Genotoxicity and

Immunotoxicity evaluations were exposed for 6 hours/day,

generally 5 days per week for 4 weeks for at least 20 exposures.

Test substance administration continued through the day prior to

necropsy for all animals except th e animals in Groups I and IV

designated for recovery which were held untreated for at least

4 additional weeks prior to necropsy.

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2.11.5. ADMINISTRATION OF TEST SUBSTANCE

The test substance was administered as a vapor in the breathing air

of the animals. The test atmosphere was generated by an

appropriate procedure determined during pre-study trials. The

trials were performed to evaluate the optimal set of conditions and

equipment to generate a stable atmosphere at the target exposure

levels and maintain uniform conditions throughout the exposure

chambers. Two chambers (designated chambers A and B) were used

for each exposure level, in general one for male animals and one for

female animals.

Group I

Houseline nitrogen was delivered from a regulator with a

backpressure gauge via 1⁄4” tubing to a flowmeter regulated by a

metering valve. This nitrogen flow (~ 20 Lpm) was then split via a

1⁄4” stainless steel “T” to bot h chamber turrets of the 1m 3 

# glass and

stainless steel exposure chambers where it was mixed with room

air as it was drawn into the chambers.

Groups II, III and IV

Houseline nitrogen was delivered from a regulator with a

backpressure gauge through a stainl ess steel fitting to create three

flow systems: the test substance pressurization flow, the purge

flow and the volatilization flow.

The nitrogen for the test substance pressurization flow was

directed through a metering valve, attached to a backpressure

gauge, into the vapor inlet valve of the test substance cylinder.

The metering valve was used to adjust and maintain the pressure

within the cylinder. From the pressurized cylinder, the test

substance flowed from the liquid outlet valve through a disconnect

fitting and through a filter to prev ent equipment contamination.

From the filter, the test substance flowed to a liquid flowmeter via

1 

# / 8 

# ” tubing. The outlet of the flowmeter was regulated by a

metering valve. From this metering valve, the test substance

flowed via 1 

# / 8 

# ” tubing onto the glass helix of a counter current

volatilization chamber. This glass helix was heated by a nichrome

wire which was controlled by an autotransformer and was inserted

in the center of the glass tube that supported the helix. The

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targeted temperature within the counter current chamber for each

group (83 ° F for Group II, 80 ° F for Group III and 78 ° F for Group

IV) was monitored using a digital alarm module.

The nitrogen for the purge flow system was directed, via 1⁄4” tubing

to a flowmeter regulated by a metering valve. The purge nitrogen

was delivered via 1 

# / 8 

# ” tubing to the bottom of the tube containing

the nichrome wire. This nitr ogen flow continuously purged the

area surrounding the nichrome wire within the tube, thereby,

protecting the wire from oxidation.

The nitrogen for the volatilization system was directed via 1⁄4”

tubing to a flowmeter regulated by a metering valve. From the

flowmeter, the volatilization nitrogen (~ 20 Lpm) flowed via 1⁄4”

tubing to a ball and socket joint at the bottom of the volatilization

chamber. This nitrogen flowed up through the volatilization

chamber passing over the coil and volatilizing the test substance.

The pressure within the counter-current volatilization chamber was

maintained slightly negative to the room and was monitored with a

pressure gauge.

This test substance laden nitrogen exited the top of the

volatilization chamber through a stainless steel “T” which divided

the flow, via 1⁄2” tubing, to the turrets of two 1m 3 

# glass and

stainless steel exposure chambers. As the test substance laden

nitrogen was drawn into each of the chambers, it was mixed with

room air.

The whole-body exposure chambers each had a volume of

approximately 1000 Liters (1m 3 

# ). Each chamber was operated at a

minimum flow rate of 200 Liters per minute. The final airflow

was set to provide at least one air change (calculated by dividing

the chamber volume by the airflow rate) in 5.0 minutes (12 air

changes/hour) and a T 99 

# equilibrium time (calculated by

multiplying the air change by the expone ntial factor 4.6) of at most

23 minutes.

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Grou p Airflow Rate Air Chan g e T 99



# ( L p m ) ( min ) ( min )

Chambers A/B Chambers A/B Chambers A/B

I 232/224 4.3/4.5 20/21

II 227/221 4.4/4.5 20/21

III 227/227 4.4/4.4 20/20

IV 224/224 4.5/4.5 21/21

This chamber size and airflow rates were considered adequate to

maintain the oxygen level at least 19% and the animal loading

factor below 5%. At the end of each exposure, all animals

generally remained in the chambe r for a minimum of 30 minutes.

During this time, each chamber was operated at approximately the

same flow rate using clean air only. The chambers were exhausted

through the in house filtering system , which consisted of a coarse

filter, a HEPA filter and activated charcoal.

See Figure 1 and Appendix N for equipment details.

2.11.6. EXPOSURE CONCENTRATION DETERMINATION

A nominal exposure concentration wa s calculated. The flow of air

through the chamber was monitored using appropriate calibrated

equipment. The test substance consumed (weight difference of the

5 gallon cylinder) during the exposure (mg) was divided by the

total volume of air (m 3 

# ) passing through the chamber (volumetric

combined flow rate for the 2 ch ambers times total exposure time)

to calculate the nominal concentration (mg/m 3 

# ).

During each exposure, measurements of airborne concentrations were

performed in the animals’ breathing zone at least 4 times using an

appropriate sampling procedure (~20 Lpm) and infrared (IR)

spectrophotometric analytical proce dure. Also, one charcoal tube

sample was collected (0.2 Lpm) per chamber per week and analyzed

by gas chromatography (GC) to characterize at least 10 major

components (comprising at least 80% by weight of the test substance)

to show test substance stability and comparison between the neat

liquid test substance and the vaporized test atmospheres.

See Appendices N and O for equipment details. See Appendix S

for Analytical Report.

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2.11.7. PARTICLE SIZE DISTRIBUTION ANALYSIS

During each week of exposure, par ticle size determinations were

performed using a TSI Aerodynamic Particle Sizer to characterize

the aerodynamic particle size distri bution of any aerosol present.

The samples were drawn for 20 seconds at a flow rate of 5.00

Lpm. The mass median aerodynamic diameter, geometric standard

deviation and total mass concentration were calculated based on

the amount of particles collected.

See Appendix N for equipment details.

2.11.8. CHAMBER AND EXPOSURE ROOM ENVIRONMENT

Chamber oxygen levels (maintained at least 19%) were measured

pretest and at the beginning and end of the study. Air samples in

the exposure room were taken in the vapor generation area pretest

and at the beginning and end of the study. Light (maintained

approximately 30-40 foot-candles at 1.0 meter above the floor) and

noise levels (maintained below 85 decibels) in the exposure room

were measured pretest and at the beginning and end of the study.

See Appendix N for equipment details.

2.12. EXPERIMENTAL EVALUATIONS

2.12.1. VIABILITY CHECKS (IN-CAGE)

Animals were observed in their cages twice daily for general

appearance, mortality and signs of severe toxic or pharmacologic

effects.

2.12.2. PHYSICAL EXAMINATIONS

All animals were observed as a group at least once during each

exposure.

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Each animal (except Neuropathology, GFAP, Genotoxicity and

Immunotoxicity animals) was removed from its cage and examined

twice pretest and once weekly during the study period.

Examinations included observations of general condition, skin and

fur, eyes, nose, oral cavity, abdomen and external genitalia as well

as evaluations of respiration, palpation for tissue masses,

circulatory effects, autonomic effects, central nervous system

effects, changes in motor activity, and reactivity to handling or

sensory stimuli. Neuropathology, GFAP, Genotoxicity and

Immunotoxicity animals were removed from their cage and

examined once pretest before selection for the study.

2.12.3. OPHTHALMOSCOPIC EXAMINATION

All animals (except Genotoxicity and Immunotoxicity animals) were

examined pretest and at study termination. Eyelids, lacrimal

apparatus and conjunctiva were examined grossly; cornea, anterior

chamber, lens, vitreous humor, retina and optic disc were examined

by indirect ophthalmoscopy. The eyes were examined after

instillation of a mydriatic (Mydriacyl ® 

# 1%).

2.12.4. BODY WEIGHT

Animals were removed from their cages and weighed twice

pretest, weekly during treatment and terminally (after fasting).

Terminal, fasted body weights were obtained just prior to

necropsy.

2.12.5. FEED CONSUMPTION

Feed consumption was measured (w eighed) during the week prior to

treatment initiation and weekly throughout the study. Feed was

available without restriction 7 days/week except during inhalation

exposures and when fasting prior to blood collection. Animals

were presented with weighed feeder s at the scheduled intervals.

After 7 days, the feeders were re weighed and the resulting weight

subtracted from the initial feeder weight to obtain the grams of

feed consumed per animal per week. The grams consumed per

kilogram of body weight per day was then calculated for each

animal.

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Calculation

Feed Consumption (g/kg/day) = grams of feed consumed ÷ 7 days

body weight (kg) a



# a 

# Average of current and prior measurement

2.13. NEUROBEHAVIORAL STUDIES

Testing was staggered over four se ssions with approximately equal

numbers of animals per sex per expos ure group in each session. Animals

being tested were not exposed on days of neurological evaluations, and

testing was conducted at least 16 hours after the previous exposure.

Testing was performed on the Neuropa thology and the GFAP animals.

Noise level in the functional observational battery room was maintained

within a level of 55 to 65 decibels by a white noise generator, Model MDF

5001 (MDF Products, Inc. Danbury, Connecticut). Temperature,

humidity, noise level and illumination of each room were measured and

recorded to ensure that variations in environmental conditions were

minimal during all evaluations. The functional observational battery was

performed for all animals before evaluation of motor activity.

2.13.1. FUNCTIONAL OBSERVATIONAL BATTERY

Method

A functional observational battery (Moser, 1989) was performed

on 10 animals/sex/group. With the exception of pretest,

evaluations were performed “blind”, i.e., the observer did not

know the identity of the animal’s exposure group. Time of testing

was balanced across treatment groups.

The following evaluations were perf ormed as part of the functional

observational battery:

Home Cage Evaluations: posture, vocalization and palpebral

closure.

Handling Evaluations: reactivity to general stimuli (such as

removal from cage or handling); assessment of signs of autonomic

function: lacrimation, salivation, altered fur appearance, or

red/crusty deposits around eyes.

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Open Field Evaluations: arousal level, locomotion and gait; count

of urination and defecation; convulsions, tremors, abnormal

movements or behaviors, excessive or repetitive actions;

piloerection and exophthalmos.

Reflex Assessments: response to visual (approach response) and

auditory (finger snap) stimuli; response to a tail pinch; pinna

reflex ; pupillary function.

Grip Strength (Meyer et al ., 1979): Grip strength was measured

using a Grip Strength Meter (Columbus Instruments International

Corporation, Columbus, Ohio).

Landing Foot Splay: Each animal was dropped into a pan of sand

from a height of one foot. The distance between the marks left by

the hindpaws was measured in centimeters.

Proprioception: Animals were restrained on a horizontal surface

by grasping the thorax. The observer gently pulled back on the

hindlimb of the rat so that the dorsal surface of the paw was on the

testing surface, and then released the hindlimb. The ability of the

animal to return the limb to a normal position was evaluated.

Air Righting Ability: Animals were held upside down and

dropped from a height of one foot into a container of bedding. The

landing position of each animal was recorded.

Body Weight: Animals were removed from their cages and

weighed using a Mettler Balance, Model PE4000 (Mettler

Instrument Corporation, Hightstown, New Jersey).

Frequency

Pretest and during the 4 th 

# , 8 th 

# and 13 th 

# weeks of exposure.

Temperature

Temperature was monitored and recorded twice daily, just prior to

the first evaluation and just after the last evaluation.

Desired: 18 to 26 ° C

Actual: 21 to 23 ° C

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Relative Humidity

Relative humidity was monitored and recorded twice daily, just

prior to the first evaluation and just after the last evaluation.

Excursions outside the specified ra nge did not affect the integrity

of the study.

Desired: 30 to 70%

Actual: 32 to 76%

Noise Level

Noise level was monitored using a Digital Sound Level Meter

(Sper Scientific, Ltd., Scottsdale, Arizona) and recorded twice

daily, just prior to the first evaluation and just after the last

evaluation.

Desired: 55 to 65 dB

Actual: 60 dB

Illumination

Illumination was monitored using a Photometer 1 (Quantum

Instruments, Inc., Garden City, New York) and recorded twice

daily, just prior to the first evaluation and just after the last

evaluation.

Desired: <80 footcandles

Actual: 54 to 57 footcandles

2.13.2. MOTOR ACTIVITY

Method

Using a modified version of Sc hulze’s procedures (Schulze, 1990),

the locomotor activity of 10 animals/sex/group was monitored

using an automated Photobeam Activity System (San Diego

Instruments, Inc., San Diego, California). Sessions were

60 minutes in length; each session was divided into 12 intervals of

5 minutes. The time of testing was balanced across treatment

groups.

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Frequency

Pretest and during the 4 th 

# , 8 th 

# and 13 th 

# weeks of exposure.

Temperature

Temperature was monitored and recorded twice daily, just prior to

the first evaluation and just after the last evaluation.

Desired: 18 to 26 ° C

Actual: 22 to 24 ° C

Relative Humidity

Relative humidity was monitored and recorded twice daily, just

prior to the first evaluation and just after the last evaluation.

Excursions outside the specified ranges were not considered to

have affected the integrity of the study.

Desired: 30 to 70%

Actual: 28 to 72%

Noise Level

Noise level was monitored using a Digital Sound Level Meter,

Model 840029 (Sper Scientific, Ltd., Scottsdale, Arizona) and

recorded twice daily, just prior to the first evaluation and just after

the last evaluation.

Desired: 55 to 65 dB

Actual: 60 to 62 dB

Illumination

Illumination was monitored using a Photometer 1 (Quantum

Instruments, Inc., Garden City, New York) and recorded twice

daily, just prior to the first evaluation and just after the last

evaluation.

Desired: <80 footcandles

Actual: 56 to 59 footcandles

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2.14. CLINICAL PATHOLOGY

Clinical pathology procedures and parameters were based on those

recommended in guidelines published by the Joint Scientific Committee

for International Harmonization of Clinical Pathology Testing in

“Harmonization of Animal Clinical Pathology Testing in Toxicity and

Safety Studies”, Fund. Appl. Tox. : 29, 198-201 (1996).

Blood were obtained from lightly anesthetized (carbon dioxide/oxygen

; 

# 60%/40%) animals via puncture of the orbital sinus (retrobulbar). Rats were

fasted overnight prior to blood collection. The Main Study animals

(10/sex/group) were bled at the Interim (~4 weeks) and Terminal intervals.

Groups I & IV animals in Recovery ( 10/sex/group) were bled for 

# hematology

and clinical chemistry only.

2.14.1. HEMATOLOGY

Blood for hematology studies was collected (approximately

0.25 mL) into tubes containing EDTA anticoagulant.

Blood samples were analyzed as follows:

ADVIA 120 HematologyAnalyzer, Bayer Corporation.

Hemoglobin concentration

Hematocrit

Erythrocyte count

Platelet count

Mean corpuscular volume

Mean corpuscular hemoglobin

Mean corpuscular hemoglobin concentration

Total leukocyte count

Reticulocyte count

Differential leukocyte count 1



# Other

Erythrocyte morphology (Henry, 1991)

1 

# When questionable values were obtained, manual differential leukocyte counts (Henry, 1991) and

absolute value calculations were performed for verification.

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2.14.2. COAGULATION

Blood for coagulation studies was collected (approximately

1.0 mL) into tubes containing sodium citrate anticoagulant.

Blood samples were analyzed as follows:

Mechanical clot detection system, STA Compact ®



# , Diagnostica Stago Products

Prothrombin time

Activated partial thromboplastin time

2.14.3. CLINICAL CHEMISTRY

Blood for clinical chemistry studies was collected (approximately

1.5 mL for all rats except ~1.0 mL for the female rats at the 4 th



# week interval) into tubes with no anticoagulant, allowed to clot,

and centrifuged to obtain serum.

Blood samples were analyzed as follows:

Hitachi 717, Roche Corporation Automatic Analyzer

Aspartate aminotransferase ( Kinetic - Modified IFCC Technique )



# Alanine aminotransferase ( Kinetic - Modified IFCC Technique )



# Alkaline phosphatase ( Kinetic – Modified AMP Buffer )



# Lactate dehydrogenase ( Kinetic - Lactate-pyruvate Technique )



# Sorbitol dehydrogenase ( Sigma Diagnostics NADH )



# Blood urea nitrogen ( Kinetic - Modified Urease )



# Creatinine ( Kinetic –Modified Jaffe Method )



# Glucose ( Hexokinase Method )



# Creatine kinase ( Kinetic – Modified NAC Method )



# Cholesterol ( Enzymatic – Modified Trinder Method )



# Total protein ( Biuret Technique )



# Albumin ( Bromocresol Green Method )



# Total bilirubin ( Modified Wahlefield et al. )



# Direct bilirubin ( Modified Jendrassik and Grof Method )



# Sodium ( Ion Selective Electrode )



# Potassium ( Ion Selective Electrode )



# Chloride ( Ion Selective Electrode )



# Calcium ( Cresolphthalein Complexone Method )

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Inorganic phosphorus ( Phosphomolybdate - UV Method )



# Gamma-glutamyl transferase ( Kinetic – Modified Persijn and Vander Silk

Method)



# Bile acids ( Mashige, et al., Enzymatic Method, Sigma )



# Other

Globulin ( calculated value; total protein - albumin )



# Albumin/globulin ratio ( calculated value; albumin ÷ globulin )



# Indirect bilirubin ( calculated value; total bilirubin – direct bilirubin )



# 2.14.4. RETENTION/STORAGE OF SPECIMENS

Any remaining (frozen) serum, which may have limited storage

stability, will be stored for up to six months after completion of

assays and will then be discarded. Peripheral blood smears will be

retained and archived with the study.

2.15. GENOTOXICITY EVALUATIONS

Note: The same animals were used for both of the following evaluations

except a separate positive control group was used for each evaluation:

Sister Chromatid Exchange (SCE) Evaluations

Sprague Dawley-derived (CD  

# ) rats, 5/sex/group, were exposed to

Gasoline DIPE Vapor Condensate via i nhalation for a 4 week (5 days per

week) exposure period by Huntingdon Life Sciences (PRC). PRC

personnel also performed the treatme nt of non-exposed (5/sex) positive

control animals (5 mg/kg cyclophosphamide, IP with a 0.5 mg/mL

solution @ 10 mL/kg, within 24 hours pr ior to sacrifice). BioReliance

personnel sacrificed (isoflurane inhalation) the animals on the day after

the final exposure, collected blood (2-4 mL placed into sodium heparin

tubes) from the abdominal aorta and transported the samples on ice packs

to BioReliance, Rockville, MD (attn: Ramadevi Gudi, Ph.D.). Upon

receipt, samples were processed and evaluated for determination of SCE

response as detailed in the referenced guidelines and procedures. The

Testing Facility was responsible for the GLP compliance of this

subcontractor.

See the BioReliance report (not included within this report) for materials

& methods, results, discussion and conclusion.

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Micronucleus Evaluations

Sprague Dawley-derived (CD  

# ) rats, 5/sex/group, were exposed to

Gasoline DIPE Vapor Condensate via i nhalation for a 4 week (5 days per

week) exposure period by Huntingdon Life Sciences (PRC). PRC

personnel also performed the treatme nt of non-exposed (5/sex) positive

control animals (40 mg/kg cyclophosphamide, IP with a 4.0 mg/mL

solution @ 10 mL/kg, within 24 hours prior to sacrifice). The positive

control animals were sacrificed under carbon dioxide anesthesia.

BioReliance personnel removed the femurs immediately after the blood

sample collection for the SCE evaluation and prepared fixed, unstained

slides which PRC personnel shipped to Eye Research Centre, Eye,

Suffolk, UK (attn: Christine Mason, B.Sc.). Upon receipt, the slides were

stained and evaluated for determination of micronucleus response as

detailed in the referenced guidelines and procedures. The Testing Facility

was responsible for the GLP compliance of this subcontractor.

See the Huntingdon UK report (not included within this report) for

materials & methods, results, discussion and conclusion.

2.16. IMMUNOTOXICITY EVALUATIONS

Female Sprague Dawley-derived (CD  

# ) rats, 10/group, were exposed to

Gasoline DIPE Vapor Condensate via i nhalation for a 4 week (5 days per

week) exposure period by Huntingdon Life Sciences (PRC). PRC

personnel also performed the treatment of non-exposed (10 females)

positive control animals (50 mg/kg cyclophosphamide, IP with a 5 mg/mL

solution @ 10 mL/kg, once per day for four days prior to sacrifice). Four

days prior to sacrifice, animals were sensitized with sheep red blood cells

(2 x 10 8 

# ) via tail vein injection by ImmunoTox, Inc. personnel. On the

day after the final exposure, bl ood samples were obtained from

anesthetized (carbon dioxide/oxygen) an imals via orbital collection, and

the serum separated and frozen at –70 o 

# C by PRC personnel. PRC

personnel then sacrificed the animal s via carbon dioxide inhalation. The

thymus of each animal was removed, weighed, and preserved (10%

neutral buffered formalin) for possible histopathology. The spleen of each

animal was aseptically removed, weighed and shipped intact in

HEPES/EBSS/Gentamicin solution (p repared at PRC) in individual

shipping containers on wet ice (2-8 o 

# C) via overnight delivery to

ImmunoTox., Inc., FedEx Station 3 South, 10 th 

# Street, Richmond, VA

(attn: Vanessa L. Peachee, M.S.). Upon receipt, spleen samples were

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further processed for determination of Plaque Forming Cell (PFC)

response as detailed in the referenced guidelines and procedures. The

frozen serum samples will be retain ed by the Testing Facility until the

spleen samples are evaluated for all subchronic inhalation studies in this test

program and then discarded if not need ed by the subcontractor. The Testing

Facility was responsible for the GLP compliance of this subcontractor.

See the Immunotox report (not included within this report) for materials &

methods, results, discussion and conclusion.

2.17. POSTMORTEM EVALUATIONS

2.17.1. NECROPSY INFORMATION

Necropsy of all animals was perf ormed after having been treated

for 13 weeks and/or held for appr oximately 4 weeks of recovery.

Animals were fasted overnight prior to necropsy. A necropsy

schedule was established to ensure that approximately equal

numbers of males and females were examined on each day of

necropsy and that examination of animals of both sexes were

performed at similar times of the day throughout the necropsy

period.

Method of Euthanasia

Exsanguination following carbon di oxide inhalation for Main

Study and Recovery Study animals.

2.17.2. MACROSCOPIC EXAMINATIONS

Complete macroscopic postmortem examinations were performed

on all Main/Recovery and Neuropathology animals. The GFAP

animal (Animal No. 2070) that wa s sacrificed for humane reasons

prior to termination was also subject to a macroscopic

examination. The macroscopic postmortem examinations included

examination of the external surface and all orifices; the external

surfaces of the brain and spinal co rd; the organs and tissues of the

cranial, thoracic, abdominal and pelvic cavities and neck; and the

remainder of the carcass for the presence of macroscopic

morphologic abnormalities.

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2.17.3. ORGAN WEIGHTS

Organs indicated in Table I (page 39) were weighed for all Main

and Recovery Study animals at the scheduled necropsy. Prior to

weighing, the organs were carefully dissected and properly

trimmed to remove adipose and other contiguous tissues in a

uniform manner. Organs were we ighed as soon as possible after

dissection in order to avoid drying . Paired organs were weighed

together.

2.17.4. TISSUES PRESERVED AND EXAMINED HISTOPATHOLOGICALLY

The tissues listed in Table I (p age 39) were obtained at the

scheduled sacrifice interval and preserved for all Main and

Recovery Study animals. In addition, slides of the indicated

tissues were prepared and examined microscopically for Main

Study and Recovery (gross lesions only) animals. Any

abnormalities not noted during macroscopic examinations which

were seen during histology processing were recorded.

Screening for alpha 2u-globulin nephropathy

Slides were prepared from tissu e samples of the kidneys of all

Groups I and IV main study male animals from the terminal

necropsy, using hematoxylin and eo sin stain only, not with Mallory

Heidenhain stain.

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TABLE I

TISSUE WEIGHED PRESERVED EXAMINED

MICROSCOPICALLY

Groups

I, IV II, III

Adrenal glands X X X

Aorta (thoracic) X X

Bone (sternum/femur with articular

surface) X X

Bone marrow (femur) X a a

Brain (medulla/pons, cerebrum and

cerebellum) X X X

Epididymides X X X

Esophagus X X

Eyes with optic nerve X

Heart X X X

Kidneys X X X

Lacrimal glands X

Large intestine (cecum, colon and rectum) X X

Larynx b 

# X X

Liver X X X

Lungs (with mainstem bronchi) X X X X

Lymph nodes (mediastinal and mesenteric) X X

Mammary gland X

Muscle (biceps femoris) X

Nasopharyngeal tissue c 

# X X

Nerve (sciatic) X X

Ovaries X X X

Pancreas X X

Pituitary X

Prostate X X X

Salivary glands with submandibular lymph

Node X X

Seminal vesicles X X X

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TISSUE WEIGHED PRESERVED EXAMINED

MICROSCOPICALLY

Groups

I, IV II, III

Skin X

Small intestine (duodenum, ileum,

jejunum) X X

Spinal cord (cervical, thoracic, lumbar) X X

Spleen X X X

Stomach X X

Testes X X X

Thymus X X X

Thyroid (with parathyroids) X X

Trachea X X

Urinary bladder X X

Uterus (body/horns with cervix) X X X

Zymbal’s gland X

Gross lesions and tissue masses X X X

a 

# No examination; bone marrow smears were prepared and archived.

b 

# The laryngeal mucosa was examined. Sections of the larynx examined included the epith elium covering the

base of the epiglottis, the ventral pouch and the medial surfaces of th e vocal processes of the arytenoid

cartilages.

c 

# Four sections of the nasopharyngeal tissue were examin ed. This included sections through the nasal cavity

and examinations of the squamous, transitional, respiratory and olfactory epithelia.



# Preservatives

All tissues - 10% neutral buffered formalin.

Eyes were placed in glutaraldehyde/paraformaldehyde initially and

then retained in 10% formalin. Testes and epididymides were

placed in Modified Davidson’s solution initially and then retained

in 10% formalin. Lungs and urin ary bladder were infused with

formalin prior to their immersion into a larger volume of the same

fixative.

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Processing

After fixation, the tissues and organs from all animals were

routinely processed, embedded in paraffin, cut at a microtome

setting of 4-7 microns, mounted on glass slides, stained with

hematoxylin and eosin and examined by light microscopy. The

bones were decalcified in Decalcifier II TM

.

After decalcification the skull was serially sectioned transversely

at approximately one centimeter intervals. All sections were

examined, post-fixation, for the presence of macroscopically

visible morphologic abnormalities. Four sections per animal,

described as follows, were processe d, embedded in paraffin, cut at

4-7 microns, mounted on glass slides, stained with hematoxylin

and eosin and examined by light microscopy. The first section

included the area between the upper incisor tooth and incisive

papilla. The second section include d the area between the incisive

papilla and the first palatal ridge. The third section included the

area between the second palatal ridge and first upper molar tooth.

The fourth section included the area between the first upper molar

tooth and nasopharynx.

Larynx sections were prepared from two sites; one was the area of

the ventral diverticulum and the other was the area of the ventral

seromucous glands at the base of the epiglottis. These were

classified as Larynx: Ventral Diverticulum (V-DVTC) and

Larynx: Ventral Seromucous Glands (V-SM-G), respectively, for

the purposes of data entry. In a few instances, sections of larynx

were not from the aforementioned pl anes of section. These were

classified simply as Larynx for the purposes of data entry.

2.18. NEUROPATHOLOGY SACRIFICE

Following 13 weeks of exposures, designated animals (5/sex/group) were

anesthetized with an intraperitoneal injection (~1.5 mL/kg or

~0.6 mL/male and ~0.4 mL/female) of ketamine/xylazine/acepromazine

and transcardially perfused with phosphate buffered saline followed by

1% glutaraldehyde and 4% paraformaldehyde in the same buffer. After

perfusion, the tissues listed in Table II below were obtained.

Measurement of the size (length and wi dth) and weight of the whole brain

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(cerebrum, cerebellum and pons-medulla) were made. All tissues were

then placed into a fresh solution of the same fixative prior to processing.

TABLE II

Tissue Preserved

(Groups) Microscopic

Examination

Groups

All I and IV II and III

brain (forebrain, central cerebrum, hippocampus, basal

ganglia, midbrain, cerebellum and pons, medulla) X X

eye with optic nerve X X

spinal cord (cervical, t horacic, lumbar, cross and

longitudinal sections) X X

sciatic nerve (cross and longitudinal sections) X X

tibial nerve (cross and longitudinal sections) X X

sural nerve (cross and longitudinal sections) X X

trigeminal ganglia X X

dorsal root ganglia (from C 3 

# -C 6 

# and L 4 

# -L 6 

# ) X X

dorsal root fibers (from C 3 

# -C 6 

# and L 4 

# -L 6 

# ) X X

ventral root fibers (from C 3 

# -C 6 

# and L 4 

# -L 6 

# ) X X

lungs and trachea X X X

tissues with macroscopic findings X X X



# Peripheral nerves were post-fixed in 1% osmium tetroxide,

processed and embedded in epoxy resin and sectioned at

approximately 2 microns and stained with toluidine blue. All other

tissues, including the brain, eye with optic nerve, spinal cord,

trigeminal ganglia, dorsal root ganglia, dorsal and ventral root

fibers, lungs and trachea were pr ocessed by standard techniques,

embedded in paraffin and sectioned at approximately 6 microns.

The tissues listed in Table II above were examined microscopically

for all animals as indicated. Tissues with macroscopic lesions

were examined in all animals. Any abnormalities not noted during

macroscopic postmortem examinations, which were seen during

histological processing, were recorded.

2.19. GFAP EVALUATIONS

Sprague Dawley-derived (CD  

# ) rats, 5/sex/group, were exposed to

Gasoline DIPE Vapor Condensate via i nhalation for a 13 week (5 days per

week) exposure period by Huntingdon Life Sciences (PRC). United States

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Center for Disease Control (CDC) Health Effects Laboratory personnel

sacrificed (via carbon dioxide inhalation) the animals after the 13 week

exposure period, and removed, weighe d and processed the brains. PRC

personnel shipped the samples on dry ice to the CDC Health Effects

Laboratory, Morgantown, WV (attn: James P. O’Callaghan, Ph.D.). Upon

receipt, samples were further processed for determination of GFAP

response as detailed in the referenced guidelines and procedures. The

Sponsor was responsible for the sp irit of GLP compliance of this

subcontractor.

See the CDC report (not included with in this report) for materials &

methods, results, discussion and conclusion.

2.20. STATISTICAL ANALYSIS

The following parameters were analyzed statistically:

mean body weight values and body weight changes (from pretest)

mean feed consumption values (pre sented as grams of feed/kg of body

weight/day)

mean clinical pathology values

mean organ weights, organ/body wei ght ratios and organ/brain weight

ratios

mean motor activity counts

mean FOB data including forelimb and hindlimb grip strength

measurements and mean landing foot splay measurements

2.20.1. METHOD OF ANALYSIS

Mean values of all exposure groups were compared to the mean

value for the control group at each time interval.

Evaluation of equality of group means was made by the

appropriate statistical method, followed by a multiple comparison

test if needed. Bartlett’s test (Bartlett, 1937; Sokal and Rohlf,

1995) was performed to determine if groups had equal variances.

For all parameters except organ we ights, if the variances were

equal, parametric procedures were used; if not, nonparametric

procedures were used. Organ we ight data was analyzed only by

parametric methods. The parametric method was the standard oneway analysis of variance (ANOVA) using the F ratio to assess

significance (Armitage, 1971; Dunlap and Duffy, 1975). If

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significant differences among the means were indicated, additional

tests were used to determine which means were significantly

different from the control: Dunnett’s (Dunlap et al., 1981; Dunnett,

1955, 1964), Williams (Williams, 1971, 1972), or Cochran and

Cox’s modified t-test (Cochran and Cox, 1959). The

nonparametric method was the Kruskal-Wallis test (Kruskal and

Wallis, 1952, 1953) and if differences were indicated, Shirley’s

test (Shirley, 1977), Steel’s test (Steel, 1959) or Pairwise

Comparison with Bonferroni Correction (Games and Howell,

1976) were used to determine which means differed from control.

Bartlett’s test for equality of variance was conducted at the 1%

significance level; all other statistical tests were conducted at the

5% and 1% significance levels.

Motor Activity and Functional Observational Battery (FOB) Data:

All analyses included sex as an independent variable. In those

instances where there were signifi cant effects of sex, separate

analyses by sex may have been done to explain the nature of the

effect. The analyses were perf ormed by Mark J. Nicolich, Ph.D.,

Statistician, ExxonMobil Biomedical Sciences, Inc, Annandale,

NJ. The Sponsor was responsible for the Subcontractor’s GLP

compliance.

Motor Activity Data: The analysis of the motor activity used a

mixed model analysis of covariance with an unstructured error

relationship among the 5 minute periods, and a first order

autoregressive error structure on weeks. The pretest response was

used as the covariate. The residuals from the model were tested

for normality at the p<0.01 level of significance. If the residuals

were not normally distributed, the dependent variable (number of

beam breaks) was converted to a function of its percentile rank,

then transformed by the inverse normal distribution (the

transformation is known as Blom's transformation where the rank

order is replaced by [rank-0. 375]/[n+0.25]), and reanalyzed

(Blom, 1958).

Functional Observational Data: Some of the sensorimotor and

observational battery variables were measured on a continuous

scale and some were measured on a nominal or count scale. All

measures were classified as e ither continuous or nominal. The

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analysis of the continuous variables was conducted by a mixed

model analysis of covariance with a first order autoregressive error

structure on the time points. The pr etest response was used as the

covariate. The residuals from the model were tested for normality

at the p<0.01 level of significance. Those variables that did not

exhibit normally distributed residuals at the 0.01 level were

transformed by Blom's normalized rank transformation and

reanalyzed (Blom, 1958). The nom inal data of the functional

observational battery was analyzed by a cumulative logit repeated

measures analysis with contrast terms to test for differences

between control and dosed groups (Agresti, 1989). For the

measures that were count data (s uch as number of rears or number

of urinations), the count responses may have been grouped into

several data ranges to allow sufficient data in the cells. For

example, the count of urinations might have been separated into

three groups of 0, 1 to 4, 5 or more, and analyzed.

2.21. DATA STORAGE

All raw data, preserved specimens, and retained samples, as well as the

original study protocol and the original final report are to be maintained in

the Archives of the Testing Facility (including those generated by the

subcontractors) upon completion of the study. The Sponsor will

determine the final disposition of these materials.

2.22. REGULATORY REFERENCES

2.22.1. TEST GUIDELINES

This study was designed to meet or exceed the pertinent

requirements of:

US EPA Vehicle Emissions Inha lation Exposure Guideline 79.61,

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

US EPA Subchronic Toxicity Study with Specific Health Effect

Assessments 79.62, CFR Vol. 59, No. 122, 27 June 1994.

US EPA OPPTS Health Effects Test Guidelines 870.3465, 90-Day

Inhalation Toxicity, August 1998.

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US EPA OPPTS Health Effects Test Guidelines 870.1350, Acute

Inhalation Toxicity with Histopathology (respiratory tract

histopathology sections, only), Public Draft June 1996.

US EPA OPPTS Health Effects Test Guidelines 870.6200,

Neurotoxicity Screening Battery, August 1998.

US EPA Neuropathology Assessments 79.66, CFR Vol. 59,

No. 122, 27 June 1994.

US EPA GFAP Assay 79.67, CFR Vol. 59, No. 122, 27 June 1994.

US EPA OPPTS Health Effects Test Guidelines 870.7800,

Immunotoxicity, August 1998.

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

27 June 1994.

US EPA OPPTS Health Effects Test Guidelines 870.5395,

Mammalian Erythrocyte Micronucleus Test, August 1998.

US EPA SCE Assay 79.65, CFR Vol. 59, No. 122, 27 June 1994.

2.22.2. GOOD LABORATORY PRACTICES

This study was conducted in accordance with the United States

Environmental Protection Agency’s (EPA) Good Laboratory

Practice Standards 79.60, CFR Vol. 59, No. 122, 27 June 1994,

with the exception of the characterization of the test substance,

which was not available prior to the experimental initiation.

2.22.3. ANIMAL WELFARE ACT COMPLIANCE

This study complied with all appropriate parts of the Animal

Welfare Act Regulations: 9 CF R Parts 1 and 2 Final Rules,

Federal Register, Volume 54, No. 168, August 31, 1989,

pp. 36112-36163, effective October 30, 1989 and 9 CFR Part 3

Animal Welfare Standards; Final Rule, Federal Register,

Volume 56, No. 32, February 15, 1991, pp. 6426-6505, effective

March 18, 1991.

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2.23. PROTOCOL DEVIATIONS

The following protocol deviations o ccurred during the study but were not

considered to have compromised the validity or integrity of the study:

1. Due to oversight, none of the exposure room (test substance

concentration, light and sound) a nd chamber environmental (oxygen)

parameters required to be measured at the middle of the study

(Week 6) were collected.

2. Due to fasting that night, the Group I animals were removed early

(before the thirty minute simulated clearing period was completed)

from the exposure chamber on 12 March 2002.

3. The testes from Animal Nos. 4067 and 4069 were not fixed in

Modified Davidson’s solution as required by protocol.

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3. RESULTS AND DISCUSSION

3.1. CHAMBER MONITORING

(Figure 1, Appendices A, P, Q & S)

Chamber distribution analyses (see Appendix P) showed that the test

substance was evenly distributed within each chamber. Chamber

monitoring (see Appendix Q) showed that the chamber oxygen levels

were at least 20%. Chamber room monitoring (see Appendix Q) showed

that no test substance was present in the room and that the sound and light

levels were acceptable.

The target and mean (± standard deviation) analytical (IR) and nominal

concentrations (see Appendix A) are summarized as follows:

Group Test

Substance Target

Concentration

(mg/m 3 

# ) Analytical

Concentration*

(mg/m 3 

# ) Nominal

Concentration*

(mg/m 3 

# )

I Air Control 0 0 ± 0 0 ± 0

II Gasoline DIPE

Vapor

Condensate 2000 1995 ± 132 2068 ± 83

III Gasoline DIPE

Vapor

Condensate 10000 10030 ± 692 9827 ± 284

IV Gasoline DIPE

Vapor

Condensate 20000 20120 ± 869 19070 ± 513

*These are averages for the two chambers for each exposure level since no

significant difference between chambers wa s seen. These mean and SD are for

all individual daily measurements (see Appendix A) throughout the study.

The analytically measured exposure levels of the airborne test substance

were acceptably close, in the opinion of the study director, to the targeted

exposure levels. The slight differe nces (<6%) between measured and

nominal concentrations were typical for this type of vapor exposure where

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a ratio of 1:1 is expected. Chamber environmental conditions averaged

24 ° C temperature and 50% relative humidity.

Mean particle size distribution measurements for the exposures are

summarized as follows:

Group Test

Substance Mass Median

Aerodynamic

Diameter*

(μm) Geometric

Standard

Deviation* Total

Mass

Concentration*

(mg/m 3 

# )

I Air Control 2.299 2.172 5.17 x 10 -3



# II Gasoline DIPE

Vapor

Condensate 1.861 2.036 4.15 x 10 -3



# III Gasoline DIPE

Vapor

Condensate 1.541 2.158 5.91 x 10 -3



# IV Gasoline DIPE

Vapor

Condensate 1.605 2.195 6.33 x 10 -3



# *These are averages for the two cham bers for each exposure level since no

significant difference between chambers was seen.

These results indicated that the atmospheres were essentially vapor only,

as expected, since there was no substantial difference between the test

substance chambers and the air control chambers. Some particles are

invariably present in chamber atmospheres, representing background air.

It is unlikely that gasoline vapor condensed on these particles given the

volatility of these light end vapor components and the high level of

consistency in constituent proportions demonstrated analytically from

week to week as next discussed.

Analysis of the major components in the neat test substance and the test

atmospheres (see Appendix S) showed a reasonably close comparison

between the neat test substance and th e vaporized test substance. This

data demonstrated that the test animals were exposed, as expected, to all

of the major components of the test substance in their reasonably proper

proportion. The data was consistent from week-to-week during the study

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indicating stability of the test substance and the atmosphere generation

techniques.

3.2. MORTALITY

(Appendix R)

All animals survived throughout the exposure and recovery phases of the

study except a 2000 mg/m 3 

# exposed male animal (#2070) which was

euthanized on Study Day 76 as a result of an accidental injury. Since this

animal was 1 of 5 male animals from this test group designated for GFAP

evaluation after 13 weeks of exposure, it was decided to replace this

animal with 1 of 10 male animals ( #2060) from this test group designated

for Main Study evaluations.

3.3. CLINICAL OBSERVATIONS

3.3.1. IN-CHAMBER OBSERVATIONS

(Table 1)

The test animals were unremarkable during the exposure periods.

3.3.2. WEEKLY CLINICAL OBSERVATIONS

(Table 2 & Appendix B)

An exposure-related increase in the frequency of red nasal

discharge observations was seen in the 20000 mg/m 3 

# exposed

animals compared to the Air Cont rol animals and the other test

substance exposed animals starting as early as Week 2 for the

males and Week 4 for the females. These observations abated

during the recovery period. Otherwise, the test animals were

generally unremarkable during the non-exposure periods

(afternoon evaluations). Scattered observations such as

chromodacryorrhea were noted but were not attributed to

treatment.

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3.4. OPHTHALMOSCOPIC EXAMINATIONS

(Table 3 & Appendix C)

In the opinion of the veterinary ophthalmologist, there were no indications

of treatment-related ocular disease in the test substance exposed animals.

3.5. BODY WEIGHTS

(Figures 2 - 3, Tables 4 - 5 & Appendices D - E)

There were no exposure-related differences in absolute body weights or in

body weight changes in the test substance exposed animals compared to

the Air Control animals. A few sta tistically significant increases (relative

to control values) were noted at weeks 9-12 in the 10000 mg/m 3 

# exposed

female animals compared to the Air Control animals. However, these

were not in a treatment-related pattern. Also, increases in body weights

are not usually considered an adverse response to treatment.

3.6. FEED CONSUMPTION

(Figures 4 - 5, Table 6 & Appendix F)

There were no exposure-related differences in feed consumption in the test

substance exposed animals compared to the Air Control animals. A few

statistically significant increases (relativ e to control values) were noted at

weeks 7, 8 and 10 in the 20000 mg/m 3 

# exposed female animals compared

to the Air Control animals. However, these were not in a treatmentrelated pattern. Also, increases in feed consumption are not usually

considered an adverse response to treatment. Feed consumption also

increased during the recovery period probably since the animals were no

longer in the exposure chambers without feed for 6 hours per day.

3.7. NEUROBEHAVIORAL STUDIES

3.7.1. MOTOR ACTIVITY

(Tables 7 (Summary Report) and 8 & Appendix G)

The results of the analyses indicated statistically significant

differences in responses in exposure levels for both the original

data and the Blom transformed data (p<0.002). Specifically,

contrast analysis of the Blom transformed data showed that

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response from low and medium e xposure groups were significantly

different from the control group. However, there was no linear

relationship between exposure levels and response, or any

consistent change in activity related to exposure. The attached

data plots do not suggest a consistent exposure related response,

but show an increase in activity in the female control group and

decrease in activity in the female mid exposure group only during

study Week 8 (there was not a statistically significant time by

exposure effect).

3.7.2. FUNCTIONAL OBSERVATIONAL BATTERY

(Tables 7 (Summary Report) and 9 & Appendix H)

FOB Continuous: None of the four continuous measures indicated

statistically significant exposure related differences. The residuals

from the forelimb grip strength, foot splay, and Blom transformed

hindlimb grip strength were normally distributed. The residuals

from body weight (both observed a nd Blom transformed data) and

hindlimb grip strength (observed data) were not normally

distributed. Because the signifi cance levels for the observed and

Blom transformed body weight were not near the 0.05 level of

significance the deviation from normality was not considered

crucial in these analyses.

FOB Discrete: For the 26 measures on a nominal or count scale

only 6 measures showed any variation in response: posture, ease of

removal, ease of handling, def ecation count, urination count, and

sensitivity to light touch. For these 6 measures none indicated

statistically significant exposure related differences. For the

remaining 20 variables all animals, or at most four responses (less

than 1.5% of the observations), showed the same response in all

groups at all time points and were not analyzed because of the lack

of meaningful variation in response.

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3.8. CLINICAL PATHOLOGY

3.8.1. HEMATOLOGY

(Table 10 & Appendix I)

There were exposure-related differences in hematology values in

test substance exposed animals compared to the Air Control

animals at the 4 th 

# week and terminal and recovery intervals. These

differences were limited to statistically significant decreases

(21-23%) in reticulocyte counts for the 20000 mg/m 3 

# exposed male

animals which occurred at both exposures period intervals

(4 th 

# week and terminal) and at termination of the recovery interval.

Statistically significant (13-17%) decreases also occurred in

reticulocyte counts for the 2000 and 10000 mg/m 3 

# exposed male

animals but only at the 4 th 

# week interval. Similar differences did

not occur in female animals at any exposure level and any interval

but there were statistically significant (35-36%) decreases in

absolute monocytes for the 20000 mg/m 3 

# exposed female animals

but only at the 4 th 

# week and recovery intervals. Other differences

(such as increases in platelets in all exposed groups of males at the

terminal interval) were seen but not in an exposure-level related

pattern and were attributed to normal variability.

3.8.2. COAGULATION

(Table 11 & Appendix J)

There were no exposure-related differences in coagulation values

in test substance exposed animals compared to the Air Control

animals at the 4th week and Terminal intervals. A few statistically

significant differences in prothrombin time and activated partial

thromboplastin time were noted in males at the 4th week interval

but were not in a treatment-related pattern or the absolute

differences were minimal and were not similarly seen at the

Terminal interval.

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3.8.3. CLINICAL CHEMISTRY

(Table 12 & Appendix K)

There were exposure-related differences in clinical chemistry

values in the test substance expos ed animals compared to the Air

Control animals at the 4 th 

# week and terminal and recovery

intervals. Decreases, relative to control values, occurred in several

serum enzymes for test substance-exposed groups after 4 weeks of

exposure and/or at the terminal or recovery intervals. Enzymes

affected were alanine aminotransferase, aspartate

aminotransferase, lactate dehydrogenase, creatine kinase and

sorbitol dehydrogenase. While some of the differences (e.g.

lactate dehydrogenase and sorbitol dehydrogenase) were

consistently statistically significant at the 4 weeks and terminal

and/or recovery intervals, decreases in these enzymes are not

usually considered an adverse response to treatment and there were

no corresponding histopathology findings. Other differences (such

as increases in sodium and/or potassium in all exposed groups of

females at the terminal interval) were seen but not in an exposurelevel related pattern and were attributed to normal variability.

3.9. ORGAN WEIGHTS AND BRAIN MEASUREMENTS

(Table 13 & Appendix L)

There were no exposure-related differences in organ weights and brain

measurements in the test substance exposed animals compared to the Air

Control animals at the Terminal and/or Recovery intervals. A few

statistically significant differences were noted including decreased

absolute, but not relative to body wei ght, lung weights at the Terminal

interval. However, these were not seen in an exposure level related

pattern and/or the absolute differences were minimal and there were no

corresponding histopathology findings. The brain weights in perfused

Neuropathology animals were lower in general than those in non-perfused

Main Study animals since the perfusion fixatives desiccate the brain

tissue.

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3.10. PATHOLOGY

3.10.1. MACROSCOPIC

(Table 14 & Appendix M)

No gross abnormalities related to test substance exposure were

evident at necropsy examination. The macroscopic lesions noted

were similar to those routinely encountered in animals of this age

and species or were represented by individual animals and

considered incidental findings.

3.10.2. MICROSCOPIC

(Table 15 & Appendix M)

All animals in the Air Control and 20000 mg/m 3 

# exposure groups

had all protocol-required tissues examined following the Terminal

sacrifice. The lungs and gross le sions were also examined in all

test animals following the Terminal sacrifice. No examinations

were considered necessary for the Recovery animals other than any

gross lesions. Microscopic findings that were considered

exposure-related were found only in the kidneys of male animals

exposed to 20000 mg/m 3 

# of Gasoline DIPE Vapor Condensate

following the Terminal sacrifice. Because these renal findings

were expected and have been fully investigated during prior testing

with gasoline test substances, no further examinations of animals

from the Terminal or Recovery sacrifice were made for this study.

Kidneys

Among animals having microscopic examination of complete

tissues, all male rats exposed to 20000 mg/m 3 

# of Gasoline DIPE

Vapor Condensate had eosinophilic hyaline granules within the

cytoplasm of renal proximal tubular epithelial cells. This lesion

was not noted in control animals or females exposed to the test

substance. Most 20000 mg/m 3 

# exposed males also had evidence of

tubular regeneration (basophilic tubules).

These renal histopathologic changes are consistent with hyaline

droplet nephropathy, attributable to accumulation of alpha-2

microglobulin within renal tubular epithelial cells. This species-

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and gender-specific change has been well documented in male rats

exposed to a variety of hydrocarbon compounds (Alden, CL, 1986)

and is not considered relevant to humans.

Histologic Findings in Kidneys of Rats Exposed to Gasoline

DIPE Vapor Condensate

Affected Animals

Males Females

Exposure level (mg/m 3 

# x10 3 

# ) 0 20 0 20

n= 10 10 10 10

Eosinophilic hyaline granules,

proximal tubular epithelium

Minimal 0 1 0 0

Slight 0 9 0 0

Basophilic cortical

tubules

Minimal 2 5 0 2

Slight 1 2 0 0

Other tissues: No test substance related histopathologic changes

were noted in other protocol-specified tissues, including lung,

nasoturbinates and larynx.

3.10.3. NEUROPATHOLOGY

(Table 15 & Appendix M)

No microscopic changes attributable to test substance exposure

were observed in brain, spinal cord, eyes with optic nerve,

peripheral nerves or ganglia among the 20,000 mg/m 3 

# exposed

satellite animals.

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4. CONCLUSION

Thirteen weeks of exposure of rats to Gasoline DIPE Vapor Condensate resulted

in hydrocarbon nephropathy in male animals exposed to 20000 mg/m 3 

# of vapor.

However, this finding has been ge nerally accepted (US EPA 1991. Alpha 2

microglobulin: association with chemically induced renal toxicity and neoplasia

in male rat. In Risk Assessment Fo rum, p.85. US Govt. Printing Office,

Washington, DC.) not to be relevant to hum an risk assessment. Thirteen weeks of

exposure of rats to Gasoline DIPE Vapor Condensate also resulted in increased

red nasal discharge and slight, non-reversib le decreases in reticulocyte values (at

the 4 th 

# week and terminal and recovery interv als) in the male animals exposed to

20000 mg/m 3 

# of vapor. Therefore, the 10000 mg/m 3 

# exposure level (excluding

male rat nephropathy) was considered a no observable adverse effect level

(NOAEL).

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REFERENCES

Agresti, Alan, "A survey of models for repeated ordered categorical response data",

Statistics in Medicine , vol 8, 1989, pg 1209-1224.

Alden, C.L. 1986. A Review of Uni que Male Rat Hydrocarbon Nephropathy. Toxicol

Pathol 14: 109-11.

Armitage, P. 1971. Statistical Methods in Medical Research. Oxford, UK: Blackwell

Scientific Publications.

Bartlett, M.S. 1937. Properties of sufficiency and statistical tests. Proceedings of the

Royal Society, Series A, 160: 268-282.

Blom, G., Statistical Estimates and Transforme d Beta Variables, J. Wiley and Sons, NY,

1958.

Cochran, W.G. and Cox, G.M. 1959. Experimental Designs , New York: John Wiley,

100-102.

Dunlap, W.P. and Duffy, J.A. 1975. Fortra n IV Functions for Calculating Exact

Probabilities Associated with Z, Chi-Square, T and F Values. Behav. Res. Methods and

Instrumentations 7: 59-60.

Dunlap, W.P., Marx, M.S. and Agamy, G.G. 1981. Fortran IV functions for calculating

probabilities associated with Dunnett’s test. Behav. Res. Methods and Instrumentation

13: 363-366.

Dunnett, C.W. 1955. A multiple comparison procedure for comparing several treatments

with a control. Journal of the American Statistical Association 50: 1096-1121.

Dunnett, C.W. 1964. New tables for multip le comparisons with a control. Biometrics 20-

3: 482-491.

Fisher, R.A. 1973. Statistical Methods for Research Workers . 14 th 

# edition. New York:

Hafner Publishing Company.

Galecki, A.T. “General class of covariance structures for two or more repeated factors in

longitudinal data analysis,” Communications in Statistics-Theory and Methods, 23(11),

1994 pg 3105 -3119.

# Huntingdon Life Sciences 00-6130 Page 59

211-DIPE-S Final Report

Games, P.A. and Howell, J.F. 1976. Pairwise multiple comparison procedures with

unequal n’s and/or variances: a monte-carlo study. Journal of Educational Statistics 1:

113-125.

Henry, John Bernard, M.D. 1991. Clinical Diagnosis and Management by Laboratory

Methods . 18 th 

# edition. Philadelphia: W.B. Saunders Co.

Kruskal, W.H. and Wallis, W.A. 1952. Use of Ranks in one-criterion variance analysis.

Journal of the American Statistical Association 47: 583-621.

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

American Statistical Association 48: 907-911.

Mantel, N. 1963. Chi-square test with one de gree of freedom: extensions of the MantelHaenszel procedure. Journal of the American Statistical Association 58: 690-700.

Meyer, O. et al. 1979. A method for the rou tine assessment of fore- and hindlimb grip

strength of rats and mice. Neurobehavioral Toxicology 1: 233-236.

Moser, V.C. 1989. Screening approaches to neurotoxicity: a functional observational

battery. Journal of American College of Toxicology 8-1: 85-94.

Pinheiro, J.C. and Bates, D.M. Mixed-Eff ects Models in S and S-PLUS, Springer-Verlag,

NY, 2002.

Schulze, Gene E. 1990. Large-scale assessment of motor activity in rodents: procedures

for routine use in toxicology studies. Journal of American College of Toxicology 9-4:

455-463.

Shapiro, S.S. and Wilk, M.B., “An analysis of variance test for normality (complete

samples)”, Biometrika , 52, 1965, pg 591-611.

Shirley, E.A.C. 1977. A non-parametric equivalent of Williams’ test for contrasting

increasing dose levels of a treatment. Biometrics 33: 386-389.

Sokal, R.R. and Rohlf, F.J. 1995. Biometry. 3rd Edition. San Francisco: W.H. Freeman

pp. 369-371.

# Huntingdon Life Sciences 00-6130 Page 60

211-DIPE-S Final Report

Steel, R.G.D. 1959. A multiple comparison rank sum test: Treatments versus control.

Biometrics 15: 560-572.

Williams, D.A. 1971. A test for differences between treatment means when several dose

levels are compared with a zero dose control. Biometrics 27: 103-117.

Williams, D.A. 1972. The comparison of severa l dose levels with a zero dose control.

Biometrics 28: 519-531.

# Huntingdon Life Sciences 00-6130 Page 61

211-DIPE-S Final Report

Group I Chamber Generation System

and Whole-Body Exposure Chamber Figure 1

Notes:

1. A pair of chambers was used for the Group I exposures.

2. Sampling Ports H-11 (left-bottom), H- 12 (left-top), H-13 (right-bottom) and

H-14 (right-top) used for pretest distri bution sampling, were located on the

back wall of the chambers.

# Huntingdon Life Sciences 00-6130 Page 62

211-DIPE-S Final Report

Groups II, III and IV Chamber Generation System

and Whole-Body Exposure Chamber Figure 1

Notes:

1. A pair of chambers was used for each of the Groups II, III and IV exposures.

2. Sampling Ports H-11 (left-bottom), H- 12 (left-top), H-13 (right-bottom) and

H-14 (right-top) used for pretest distri bution sampling, were located on the

back wall of the chambers.

# Huntingdon Life Sciences 00-6130 Page 63

211-DIPE-S Final Report

Males Mean Body Weights

(Main/Recovery) Figure 2

200 230 260 290 320 350 380 410 440 470 500 530 560 590

-1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17Body Weight (grams)



# Week Group I - 0 mg/m^3

Group II - 2,000 mg/m^3

Group III - 10,000 mg/m^3

Group IV - 20,000 mg/m^3

# Huntingdon Life Sciences 00-6130 Page 64

211-DIPE-S Final Report

Females Mean Body Weights

(Main/Recovery) Figure 3

170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330 340

-1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17



# WeekBody Weight (grams)



# Group I - 0 mg/m^3

Group II - 2,000 mg/m^3

Group III - 10,000 mg/m^3

Group IV - 20,000 mg/m^3

# Huntingdon Life Sciences 00-6130 Page 65

211-DIPE-S Final Report

Males Mean Feed Consumption

(Main/Recovery) Figure 4

40 50 60 70 80 90 100

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17



# WeekFeed Consumption (g/kg/day) 

# Group I - 0 mg/m^3

Group II - 2,000 mg/m^3

Group III - 10,000 mg/m^3

Group IV - 20,000 mg/m^3

# Huntingdon Life Sciences 00-6130 Page 66

211-DIPE-S Final Report

Females Mean Feed Consumption

(Main/Recovery) Figure 5

55 60 65 70 75 80 85 90 95 100

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17



# WeekFeed Consumption (g/kg/day) 

# Group I - 0 mg/m^3

Group II - 2,000 mg/m^3

Group III - 10,000 mg/m^3

Group IV - 20,000 mg/m^3

# Huntingdon Life Sciences 00-6130 Page 67

211-DIPE-S Final Report

General Preface

General Notes

When N (number of animals ) presented on the mean tables is less than that

specified in the Experimental Outline, refer to indivi dual values in appendices for

an explanation of the missing data.

Individual animal data values present ed in this report may be rounded.

Unrounded individual animal data values ar e used to calculate the reported mean

and standard deviation values. Therefore, use of the reported individual values

to reproduce means, standard deviations and/or to perform any subsequent

calculations may produce minor discr epancies between the calculated values

and those presented in this report.

Due to computer system limitation, the assigning of group numbers in some

printouts of the report was different from the protocol and other data. Groups I to

IV were presented as Groups 1 to 4 in some printouts of this report.

The following letter designations were used for some printouts for this study:

00-6130 Toxicity (Main and Recovery) Animals

00-6130N Neuropathology Animals

Key to Abbreviations

M = Males

F = Females

SD = Standard Deviation

N = Number of AnimalsHuntingdon Life Sciences01-6130Page 68211-DIPE-S FinalReportSummary of In-Chamber ObservationsTable 1All = 100% of the animals exhibiting a given observation.Note: In-chamber observations are based on all animals present in the exposure chamber at the time, including satellite animals.Exposure Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14Group I – 0 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup II – 2000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup III – 10000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup IV – 20000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllHuntingdon Life Sciences01-6130Page 69211-DIPE-S FinalReportSummary of In-Chamber ObservationsTable 1All = 100% of the animals exhibiting a given observation.Note: In-chamber observations are based on all animals present in the exposure chamber at the time, including satellite animals.Exposure Day 15 16 17 18 19 20 21 22 23 24 25 26 27 28Group I – 0 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup II – 2000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup III – 10000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup IV – 20000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllHuntingdon Life Sciences01-6130Page 70211-DIPE-S FinalReportSummary of In-Chamber ObservationsTable 1All = 100% of the animals exhibiting a given observation.Note: In-chamber observations are based on all animals present in the exposure chamber at the time, including satellite animals.Exposure Day 29 30 31 32 33 34 35 36 37 38 39 40 41 42Group I – 0 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup II – 2000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup III – 10000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup IV – 20000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllHuntingdon Life Sciences01-6130Page 71211-DIPE-S FinalReportSummary of In-Chamber ObservationsTable 1All = 100% of the animals exhibiting a given observation.Note: In-chamber observations are based on all animals present in the exposure chamber at the time, including satellite animals.Exposure Day43 44 45 46 47 48 49 50 51 52 53 54 55 56Group I – 0 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup II – 2000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup III – 10000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup IV – 20000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllHuntingdon Life Sciences01-6130Page 72211-DIPE-S FinalReportSummary of In-Chamber ObservationsTable 1All = 100% of the animals exhibiting a given observation.Note: In-chamber observations are based on all animals present in the exposure chamber at the time, including satellite animals.Exposure Day 57 58 59 60 61 62 63 64 65 66 67 68 69 70Group I – 0 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup II – 2000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup III – 10000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All AllGroup IV – 20000 mg/m3NormalWithin Normal Limits All All All AllAll All All All All All All All All All

# Huntingdon Life Sciences 00-6130 Page 73

211-DIPE-S Final Report

Summary of Clinical

Observations Table 2

For summarization purposes, descriptive comments [i.e., location of scab(s)

and/or sore(s), etc.] are not presented in this table. These data are contained in

the study raw data if needed.

Total represents a cumulative total of al l animals with the indicated observation

one or more times during the study.

Corresponding exposure levels for each group were as follows:

Group 1 – 0 mg/m 3



# Group 2 – 2000 mg/m 3



# Group 3 – 10000 mg/m 3



# Group 4 – 20000 mg/m 325-SEP-200310:42HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)MALES#OFANIMALSEXAMINEDNORMALWITHINNORMALLIMITSDEADTERMINALSACRIFICERECOVERYSACRIFICEREASSIGNEDTOSATELLITEGROUPDERMALGENERALSCABSTABLE2GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSSUMMARYOFCLINICALOBSERVATIONSDAYOFSTUDYGROUP#-11-161320273441485562697683909197104111118122TOTAL1202020202020202020202020202020210101010101010101010101010101031010101010101010101010101010104202020202020202020202020202020120201821010103101094202020123412341234123400000000000000000000000000000000000000000000201018 1816161810109 8 79 9 8 9 818 1816 14 160000000000002010000000000000101000000000000010100000000000000010000000000000001018 177 510915 150000000000000000000000000000000017 176 781015150000000000000000000000000000000015691400000000000000001669160000000000000000143413555500000000000051010101010500000500000510101010100000545500000100000090090000000000000000100010000000000000000010001000000000000000001000100200100100200010009001000100101000000001010000001000000000000002010Page 7425-SEP-200310:42HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)MALESTABLE2GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSSUMMARYOFCLINICALOBSERVATIONSDAYOFSTUDYGROUP#-11-161320273441485562 69 76839091 97104111118 122TOTAL#OFANIMALSEXAMINEDALOPECIA-GENERALALOPECIA-EXTREMITIES/SNOUTOCULARCHROMODACRYORRHEA-BILATERALCHROMODACRYORRHEA-UNILATERALORAL/BUCCALNASALDISCHARGE-RED1202020202020202020202020202020210101010101010101010101010101031010101010101010101010101010104202020202020202020202020202020123412341234123412340000000000000000000000000000000000000000000000000000000000000000000000000000000200100001000000001001001010010001000021130010110100012001210400001101000110000213000011020001100002030000110200021000141200001102000210001322000011020000100012030000110200002000231400001102000011002312000010010000000002115101050050010101000000051010101010000000000000000000000000100100000000000000000000000000000000000000000000000000000000000000000000000000000000000000000010110200023101040605011Page 7525-SEP-200310:42HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)MALESTABLE2GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSSUMMARYOFCLINICALOBSERVATIONSDAYOFSTUDYGROUP#-11-161320273441 485562 697683909197104111118122TOTAL#OFANIMALSEXAMINEDNASALDISCHARGE-CLEARINCISORSMALOCCLUDED120202101020202020202020201010101010101010310101010104202020202012341234000000000000000000000000000000000000000010101010102020202020000000010000000100000001000000010000000120101020000100012020202010101010101010102020202000000001000000010000000100000000555500000000100010000000001000100000000010001000000000100010000000001000100000000000010001Page 7625-SEP-200310:42HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)FEMALES#OFANIMALSEXAMINEDNORMALWITHINNORMALLIMITSDEADTERMINALSACRIFICERECOVERYSACRIFICEDERMALGENERALALOPECIA-GENERALALOPECIA-EXTREMITIES/SNOUTTABLE2GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSSUMMARYOFCLINICALOBSERVATIONSDAYOFSTUDYGROUP#-11-1613202734414855626976839091 97104111118 122TOTAL120202020202020202020202020202021010101010101010101010101010103101010101010101010101010101010420202020202020202020202020202012020202020210109 9 8310109 9 84202019 18 1712341234123412340000000000000000000000000000000000000000000000000000000001000000000000000100000017 17 15 16 16 15 137 8 8 8 9 8 87 5 7 6 6 68131114 14 16131200000000010010000000000001001010000000000100201000000000010020100000000001002010000000000100201000000000010030101413108 7 38 8 411119000000000100301000000000010030205555000000003010510101010105000005000005101010101000005555000000000000800800000000000010009008000000000000100090090000000000001000900902001001002000100010001000100101000000001010000010000000000001003020Page 7725-SEP-200310:42HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)FEMALESTABLE2GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSSUMMARYOFCLINICALOBSERVATIONSDAYOFSTUDYGROUP#-11-161320273441485562697683909197104111 118122TOTAL#OFANIMALSEXAMINEDYELLOWANO-GENITALSTAININGOCULARCHROMODACRYORRHEA-BILATERALCHROMODACRYORRHEA-UNILATERALLACRIMATION-UNILATERALORAL/BUCCALNASALDISCHARGE-RED1202020202020202020202020202020210101010101010101010101010101031010101010101010101010101010104202020202020202020202020202020123412341234123412340000000000000000000000000000000000000000000000000000000001110000000000000000001100000000000000000122000000000000000022360000000000010000214800000000000000003125000000010000000021350000000100000001203200010000010100003035000100000101000040270001000021010000302700020000210100003107000100001100000021055101010101050000050000051010101010000000000000000000000000000000000000100200000000000000000002000000000001000000000000000000010000000000000000000000000002000121030001070408015Page 7825-SEP-200310:42HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)FEMALESTABLE2GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSSUMMARYOFCLINICALOBSERVATIONSDAYOFSTUDYGROUP#-11-161320273441485562 69 7683909197104111118 122TOTAL#OFANIMALSEXAMINEDNASALDISCHARGE-CLEARINCISORSMALOCCLUDEDINCISORSBROKEN/MISSING12020202020202020202020202020202101010101010101010101010101010310101010101010101010101010101042020202020202020202020202020201234123412340000000000000000000000000000000000000001000000000001000000000001000000000001000100000001000000000000000100000000000100000000010100000000010100000000010100000000010100000000010000005101010101050000050000051010101010000000000000000000000000000000000001000000000001000000000001000000000000000101020001Page 7925-SEP-200311:51HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupMALES#OFANIMALSEXAMINEDNORMALWITHINNORMALLIMITSDEADTERMINALSACRIFICETABLE2GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSSUMMARYOFCLINICALOBSERVATIONSDAYOFSTUDYGROUP#-1193 94TOTAL1 53225233 5 3 24 5 2315005250053500545005103 25202353032 54023 5Page 8025-SEP-200311:51HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupFEMALES#OFANIMALSEXAMINEDNORMALWITHINNORMALLIMITSDEADTERMINALSACRIFICETABLE2GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSSUMMARYOFCLINICALOBSERVATIONSDAYOFSTUDYGROUP#-119394TOTAL15232 5323 52345 321 5005250053 500545005102 3 520325302 3 540325Page 81

# Huntingdon Life Sciences 00

- 6130 Page 82

211

- DIPE

- S Final Report

Summary of Ophthalmoscopic Findings

Preface Table 3

Table of Contents

Ophthalmologist Reports

.................................................................................... 83

Summary of Ophthalmology Observations

......................................................... 87

# Tab.le 3 - Pretest (Main Study)



# LIONEL F. RUBIN, V.M.D.

1116 Saint Andrews Road

Bryn Mawr, PA 1901 0

(61 0) 520 9430

February 2, 2002

Huntingdon Life Sciences, Inc.

Mettlers Road, Box 2360

East Millstone, NJ 08875-2360

Re: study A-12 00-6130

Ophthalmoscopic examination of study A-12 00-6130 pretest rats was performed

January 31, 2002. Rats with ocular abnormalities were identified and should be withdrawn

from inclusion in the study if convenient.

Lionel F. Rubin, V. M. D. 

# Page 83

# Table 3 - Termination (Main Study)

LIONEL F. RUBIN, V.M.D.

1116 Saint Andrews Road

Bryn Mawr, PA 19010

(61 0) 520 9430

May 11,2002

Huntingdon Life Sciences, Inc.

Mettlers Road, Box 2360

East Millstone, NJ 08875-2360

Re: study A-12 00-6130

Ophthalmoscopic examination of study A-12 00-6130 rats was performed May 10,

2002 (terminal examination). I have reviewed the findings of the type and incidence of

ocular abnormalities. There is no indication of dose or compound related ocular disease.

In my opinion, none of the ocular abnormalities is attributable to the administration of the

test compound.

Lionel F. Rubin, V. M.D. 

# Page 84

# Table 3 - Pretest (Neuropathology)

LIONEL F. RUBIN, V.M.D.

1116 Saint Andrews Road

Huntingdon Life Sciences, Inc.

Mettlers Road, Box 2360

East Millstone, NJ 08875-2360

Re: study A-12 00-6130N Bryn Mawr, PA 19010

(610) 520 9430

February 2, 2002

Ophthalmoscopic examination of study A-12 00-6130N pretest rats was performed

January 31, 2002. Rats with ocular abnormalities were identified and should be withdrawn

from inclusion in the study if convenient.

# ~~ Lionel F. Rubin, V. M. D. 

# Page 85

# Table 3 ·Termination (Neuropathology)



# LIONEL F. RUBIN, V.M.D.

Huntingdon Life Sciences, Inc.

Mettlers Road, Box 2360

East Millstone, NJ 08875-2360

Re: study A-12 00-6130N 1116 Saint Andrews Road

Bryn Mawr, PA 19010

(61 0) 520 9430

May 11,2002

Ophthalmoscopic examination of study A-12 00-6130N rats was performed May 1 0,

2002 (terminal examination). No ocular abnormalities were seen. There is no indication of

dose or compound related ocular disease.

Lionel F. Rubin, V. M.D. 

# Page 8625-SEP-200311:57HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE3GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSDOSEGROUP:DOSELEVEL(MG/M3):MALEStotalnumberexaminedNOABNORMALITIESDETECTEDNostatisticallysignificantdifferencesSUMMARYOFOPHTHALMOLOGYOBSERVATIONS1020220001031000010WEEK-242000020Page 8725-SEP-200311:57HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE3GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSDOSEGROUP:DOSELEVEL(MG/M3):FEMALEStotalnumberexaminedNOABNORMALITIESDETECTEDNostatisticallysignificantdifferencesSUMMARYOFOPHTHALMOLOGYOBSERVATIONS1020220001031000010WEEK-242000020Page 8825-SEP-200311:57HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE3GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSDOSEGROUP:DOSELEVEL(MG/M3):MALEStotalnumberexaminedCONJUNCTIVANCONJUNCTIVITISN%NostatisticallysignificantdifferencesSUMMARYOFOPHTHALMOLOGYOBSERVATIONS1020000.022000101110.031000010000.0WEEK1242000020115.0Page 8925-SEP-200311:57HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE3GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSDOSEGROUP:DOSELEVEL(MG/M3):FEMALEStotalnumberexaminedIRISNANTERIORUVEITIS(IRITIS)N%CONJUNCTIVAN-----------CONJUNCTIVITISN%NostatisticallysignificantdifferencesSUMMARYOFOPHTHALMOLOGYOBSERVATIONS1020000.0115.02200010000.01110.031000010000.0000.0WEEK1242000020115.0115.0Page 9025-SEP-200311:59HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE3GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSDOSEGROUP:DOSELEVEL(MG/M3):MALEStotalnumberexaminedNOABNORMALITIESDETECTEDNostatisticallysignificantdifferencesSUMMARYOFOPHTHALMOLOGYOBSERVATIONS1052200053100005WEEK-24200005Page 9125-SEP-200311:59HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE3GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSDOSEGROUP:DOSELEVEL(MG/M3):FEMALEStotalnumberexaminedNOABNORMALITIESDETECTEDNostatisticallysignificantdifferencesSUMMARYOFOPHTHALMOLOGYOBSERVATIONS1052200053100005WEEK-24200005Page 9225-SEP-200311:59HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE3GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSDOSEGROUP:DOSELEVEL(MG/M3):MALEStotalnumberexaminedNOABNORMALITIESDETECTEDNostatisticallysignificantdifferencesSUMMARYOFOPHTHALMOLOGYOBSERVATIONS1052200053100005WEEK124200005Page 9325-SEP-200311:59HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE3GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSDOSEGROUP:DOSELEVEL(MG/M3):FEMALEStotalnumberexaminedNOABNORMALITIESDETECTEDNostatisticallysignificantdifferencesSUMMARYOFOPHTHALMOLOGYOBSERVATIONS1052200053100005WEEK124200005Page 9425-SEP-200312:00HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE4GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK-1MEANS.D.NWEEK0MEANS.D.NWEEK1MEANS.D.NWEEK2MEANS.D.NWEEK3MEANS.D.NWEEK4MEANS.D.NWEEK5MEANS.D.NWEEK6MEANS.D.NWEEK7MEANS.D.NMEANBODYWEIGHTS(GRAMS)102208.12030613.72034218.52037319.62039919.82042022.62043922.42045523.72047925.920220002207.51030710.41034211.01037312.01040213.91042716.91044318.01046220.21048222.7103100002208.11030718.41033724.91036926.31039128.31041729.11043531.61045532.11047933.4104200002208.12030314.52033520.12036422.92039025.52041129.32042931.72044733.02047236.420-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 9525-SEP-200312:00HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE4GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK8MEANS.D.NWEEK9MEANS.D.NWEEK10MEANS.D.NWEEK11MEANS.D.NWEEK12MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTS(GRAMS)1049226.52050727.32051827.52053327.62054228.5202200049723.81051125.81051926.51053229.71053635.61031000049434.61050738.31052241.31053343.21054045.21042000048437.22050239.72051441.12052645.42053446.620Page 9625-SEP-200312:01HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE4GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK13MEANS.D.NWEEK14MEANS.D.NWEEK15MEANS.D.NWEEK16MEANS.D.NWEEK17MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTS(GRAMS)1053833.21054633.81055631.91056535.71056935.91042000052732.31053930.71055430.61056431.31057029.610Page 9725-SEP-200312:03HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE4GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK-1MEANS.D.NWEEK0MEANS.D.NWEEK1MEANS.D.NWEEK2MEANS.D.NWEEK3MEANS.D.NWEEK4MEANS.D.NWEEK5MEANS.D.NWEEK6MEANS.D.NWEEK7MEANS.D.NMEANBODYWEIGHTS(GRAMS)101798.62022213.42023714.12025117.72025918.22027017.02027719.32027920.02028922.120220001798.61021714.21023616.21024721.11025921.71026819.51027218.01027720.51028621.8103100001798.61022520.61024322.21025921.31026620.81028124.21028925.01029326.71030324.5104200001798.72022215.02023517.02024818.42025619.12026320.02027220.72027620.92028121.720-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 9825-SEP-200312:03HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE4GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK8MEANS.D.NWEEK9MEANS.D.NWEEK10MEANS.D.NWEEK11MEANS.D.NWEEK12MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTS(GRAMS)1029423.22029921.82030523.12030822.92031022.2202200029120.81029521.81030226.61030322.81030824.71031000030729.81032030.81032429.01032630.51032832.4ro42000028622.52029323.62029622.62029922.92030123.320Page 9925-SEP-200312:03HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE4GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK13MEANS.D.NWEEK14MEANS.D.NWEEK15MEANS.D.NWEEK16MEANS.D.NWEEK17MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTS(GRAMS)1030319.91030520.11030820.61030922.31030922.11042000030116.31030116.51030815.11031116.21031516.010Page 10025-SEP-200312:05HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE4GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK-1MEANS.D.NWEEK0MEANS.D.NWEEK1MEANS.D.NWEEK2MEANS.D.NWEEK3MEANS.D.NWEEK4MEANS.D.NWEEK5MEANS.D.NWEEK6MEANS.D.NWEEK7MEANS.D.NMEANBODYWEIGHTS(GRAMS)1021811.0529921.5533430.1536332.2538939.5541139.6543745.6545746.7548155.052200021615.2529824.6533534.2536939.5539541.3541946.9544349.1546154.5548059.0531000021713.6530228.4533625.0537327.2540430.1543032.0545637.3547839.7549840.1542000021810.0530621.1534523.7537925.8540528.2542727.7545030.7546338.1548138.85-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 10125-SEP-200312:05HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE4GASOLINEDIPE.VAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK8MEANS.D.NWEEK9MEANS.D.NWEEK10MEANS.D.NWEEK11MEANS.D.NWEEK12MEANS.D.NWEEK13MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTS(GRAMS)1049258.8550864.3552162.9553666.8554970.5555572.552200049559.9551362.5552359.8554160.9554862.0555569.9531000051242.8553246.6554345.6556049.8557552.8558254.0542000049048.3550651.1551858.6553160.7553870.2553770.55Page 10225-SEP-200312:06HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE4GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK-1MEANS.D.NWEEK0MEANS.D.NWEEK1MEANS.D.NWEEK2MEANS.D.NWEEK3MEANS.D.N·wEEK4MEANS.D.NWEEK5MEANS.D.NWEEK6MEANS.D.NWEEK7MEANS.D.NMEANBODYWEIGHTS(GRAMS)101837.7523015.0525213.9526822.0527619.7528622.5530025.6530228.8531228.052200018010.0522514.852449.8525710.3527319.9527620.0528715.3528818.7529821.153100001799.0522524.1523829.2525231.0525936.0527237.8527941.0528543.1529145.954200001827.752229.6524113.5525413.4526511.5527012.5528216.4528720.3529521.45-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 10325-SEP-200312:06HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE4GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK8MEANS.D.NWEEK9MEANS.D.NWEEK10MEANS.D.NWEEK11MEANS.D.NWEEK12MEANS.D.NWEEK13MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTS(GRAMS)1031629.8532131.6532635.8533033.2533738.1533240.552200029824.6530520.8531023.3531725.3532429.1531823.7531000029846.0530249.5530749.7531150.5531950.9531755.1542000029619.1530326.2530929.0531527.5531925.8531826.25Page 10425-SEP-200312:07HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO1MEANS.D.NWEEK0TO2MEANS.D.NWEEK0TO3MEANS.D.NWEEK0TO4MEANS.D.NWEEK0TO5MEANS.D.NWEEK0TO6MEANS.D.NMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)10367.5206710.6209410.82011414.02013313.82014915.02022000356.310666.610958.81012012.21013610.81015513.310310000318.4106210.2108413.91011014.31012816.51014817.610420000327.5206110.4208713.42010818.02012620.62014421.920-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 10525-SEP-200312:07HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO7MEANS.D.NWEEK0TO8MEANS.D.NWEEK0TO9MEANS.D.NWEEK0TO10MEANS.D.NWEEK0TO11MEANS.D.NWEEK0TO12MEANS.D.NMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)1017318.02018618.82020219.92021219.32022719.52023720.8202200017515.21019016.91020419.31021219.61022522.61022927.71031000017218.71018720.71020123.41021525.61022627.61023329.81042000016925.620f8126.72019929.12021130.62022334.72023135.720-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 10625-SEP-200312:09HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO13MEANS.D.NWEEK0TO14MEANS.D.NWEEK0TO15MEANS.D.NWEEK0TO16MEANS.D.NWEEK0TO17MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)1023222.71024023.71025020.91025925.01026325.51042000022722.41023920.11025421.91026423.01027022.310Page 10725-SEP-200312:11HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO1MEANS.D.NWEEK0TO2MEANS.D.NWEEK0TO3MEANS.D.NWEEK0TO4MEANS.D.NWEEK0TO5MEANS.D.NWEEK0TO6MEANS.D.NMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)10156.120299.5203710.620489.1205513.1205713.12022000196.5103012.5104111.810518.9105510.0106015.210310000189.2103411.0104113.9105611.3106414.2106915.010420000136.520267.120349.220417.920so9.2205410.120-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 10825-SEP-200312:11HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO7MEANS.D.NWEEK0TO8MEANS.D.NWEEK0TO9MEANS.D.NWEEK0TO10MEANS.D.NWEEK0TOllMEANS.D.NWEEK0TO12MEANS.D.NMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)106716.1207217.6207716.6208317.1208616.8208817.120220006912.9107412.7107814.2108519.2108515.5109116.7103100007814.8108316.51095**17.51099*17.010101*20.410103*19.3104200005911.5206410.72071ll.82074ll.42077ll.O2079ll.320-----------------------------------------------------------------------------------------------------------------------------------Statisticalkey:*=p<0.05**=p<0.01Page 10925-SEP-200312:11HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO13MEANS.D.NWEEK0TO14MEANS.D.NWEEK0TO15MEANS.D.NWEEK0TO16MEANS.D.NWEEK0TO17MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)108115.8108215.2108615.7108716.8108717.1104200007912.7107913.9108610.4109011.6109412.310Page 11025-SEP-200312:12HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO1MEANS.D.NWEEK0TO2MEANS.D.NWEEK0TO3MEANS.D.NWEEK0TO4MEANS.D.NWEEK0TO5MEANS.D.NWEEK0TO6MEANS.D.NMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)103410.256311.559019.9511122.7513828.8515830.35220003710.257115.659717.5512123.2514524.7516330.05310000358.25727.951038.5512914.5515511.9517714.25420000396.357311.059913.0512113.4514417.4515727.15-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 11125-SEP-200312:12HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO7MEANS.D.NWEEK0TO8MEANS.D.NWEEK0TO9MEANS.D.NWEEK0TO10MEANS.D.NWEEK0TO11MEANS.D.NWEEK0TO12MEANS.D.NMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)1018138.3519242.8520948.4522147.1523651.2525054.652200018234.9519736.0521538.7522537.0524337.6525038.2531000019716.2521019.4523022.3524221.3525824.2527326.2542000017529.6518442.5520044.6521252.4522555.2523365.45-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 11225-SEP-200312:16HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESWEEKDOSEGROUP:DOSELEVEL(MG/M3):0TO13MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)1025655.252200025745.6531000028026.4542000023166.25Page 11325-SEP-200312:18HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO1MEANS.D.NWEEK0TO2MEANS.D.NWEEK0TO3MEANS.D.NWEEK0TO4MEANS.D.NWEEK0TO5MEANS.D.NWEEK0TO6MEANS.D.NMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)10213.153711.654610.955611.856915.557118.6522000198.15328.25485.35518.956110.45625.55310000137.95278.953413.154715.155518.156020.254200001910.453213.054311.45499.156114.156519.05-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 11425-SEP-200312:18HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0TO7MEANS.D.NWEEK0TO8MEANS.D.NWEEK0TO9MEANS.D.NWEEK0TO10MEANS.D.NWEEK0TO11MEANS.D.NWEEK0TO12MEANS.D.NMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)108118.558622.359022.659627.259926.0510629.0522000737.857312.95809.35859.559211.659916.053100006623.657323.557726.358226.558628.159427.654200007318.657415.958123.558726.859323.659722.35-----------------------------------------------------------------------------------------------------------------------------------NostatisticallysignificantdifferencesPage 11525-SEP-200312:18HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE5GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESWEEKDOSEGROUP:DOSELEVEL(MG/M3):0TO13MEANS.D.NNostatisticallysignificantdifferencesMEANBODYWEIGHTCHANGEFROMBASELINE(GRAMS)1010231.05220009314.453100009232.054200009622.05Page 11625-SEP-200312:22HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0MEANS.D.NWEEK1MEANS.D.NWEEK2MEANS.D.NWEEK3MEANS.D.NWEEK4MEANS.D.NWEEK5MEANS.D.NWEEK6MEANS.D.NWEEK7MEANS.D.NWEEK8MEANS.D.NNostatisticallysignificantdifferencesMEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY)10974.920804.620713.420663.720623.720604.220602.920572.520532.52022000964.710773.210712.710673.010633.610591.810593.010553.510523.810310000963.710786.910714.110672.510643.19593.310615.710573.910543.210420000975.720764.420703.820664.020613.520584.720584.220563.019534.220Page 11725-SEP-200312:22HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK9MEANS.D.NWEEK10MEANS.D.NWEEK11MEANS.D.NWEEK12MEANS.D.NNostatisticallysignificantdifferencesMEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY)10522.520513.220so3.620482.72022000512.810493.010482.810463.910310000523.410512.810482.310471.610420000534.120512.820485.520482.720Page 11825-SEP-200312:23HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK13MEANS.D.NWEEK14MEANS.D.NWEEK15MEANS.D.NWEEK16MEANS.D.NWEEK17MEANS.D.NNostatisticallysignificantdifferencesMEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY)10463.010483.910483.210472.910463.510420000452.210so3.710514.310494.210483.710Page 11925-SEP-200312:23HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0MEANS.D.NWEEK1MEANS.D.NWEEK2MEANS.D.NWEEK3MEANS.D.NWEEK4MEANS.D.NWEEK5MEANS.D.NWEEK6MEANS.D.NWEEK7MEANS.D.NWEEK8MEANS.D.NNostatisticallysignificantdifferencesMEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY).10975.319813.619773.719734.120713.920673.819664.519654.820633.82022000988.010835.110776.910755.310693.59642.910664.510645.810634.610310000974.49785.89755.010713.510713.910644.510663.110653.310643.510420000969.420787.119764.720724.220704.719684.320673.820654.220643.819Page 12025-SEP-200312:23HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK9MEANS.D.NWEEK10MEANS.D.NWEEK11MEANS.D.NWEEK12MEANS.D.NNostatisticallysignificantdifferencesMEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY)10634.620623.620584.520573.82022000625.010635.410594.510613.810310000634.110614.510582.810593.110420000623.120625.420593.720604.120Page 12125-SEP-200312:24HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)TABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK13MEANS.D.NWEEK14MEANS.D.NWEEK15MEANS.D.NWEEK16MEANS.D.NWEEK17MEANS.D.NNostatisticallysignificantdifferencesMEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY)10563.810627.510583.010572.79594.110420000562.8106413.010626.19649.810604.710Page 12225-SEP-200312:25HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK0MEANS.D.NWEEK1MEANS.D.NWEEK2MEANS.D.NWEEK3MEANS.D.NWEEK4MEANS.D.NWEEK5MEANS.D.NWEEK6MEANS.D.NWEEK7MEANS.D.NWEEK8MEANS.D.NNostatisticallysignificantdifferencesMEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY)10983.85823.25733.45683.35633.55633.25603.55593.35532.4522000975.25814.45741.74672.25621.35621.55581.65551.75521.35310000962.75785.75733.35692.65641.75622.55582.15563.05521.65420000964.55793.25714.45663.75622.95623.55593.15551.65533.45Page 12325-SEP-200312:25HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK9MEANS.D.NWEEK10MEANS.D.NWEEK11MEANS.D.NWEEK12MEANS.D.NWEEK13MEANS.D.NNostatisticallysignificantdifferencesMEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY)10532.75511.15491.75492.05462.2522000522.25501.85492.15464.85441.65310000521.85502.05501.35470.65441.25420000522.05535.15491.75482.55452.25Page 12425-SEP-200312:26HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESMEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY)DOSEGROUP:DOSELEVEL(MG/M3):WEEK0MEANS.D.NWEEK1MEANS.D.NWEEK2MEANS.D.NWEEK3MEANS.D.NWEEK4MEANS.D.NWEEK5MEANS.D.NWEEK6MEANS.D.NWEEK7MEANS.D.NWEEK8MEANS.D.NStatisticalkey:*=p<0.05**=p<0.0110983.45843.55792.55744.05680.85683.45641.65632.85582.85220009710.65908.448411.75754.05683.75693.65663.55652.85593.45310000967.75795.85744.14713.55683.25652.65652.34631.15592.0542000010112.25845.55797.05765.35712.95703.55695.6568**2.4563*3.55Page 12525-SEP-200312:26HuntingdonLifeSciences00-6130NNeurotoxicitySub-GroupTABLE6GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESDOSEGROUP:DOSELEVEL(MG/M3):WEEK9MEANS.D.NWEEK10MEANS.D.NWEEK11MEANS.D.NWEEK12MEANS.D.NWEEK13MEANS.D.NStatisticalkey:*=p<0.05MEANFEEDCONSUMPTIONVALUES(GRAMS/KG/DAY)10603.05593.05583.05581.85514.3522000616.75632.55613.05603.25523.25310000591.85592.55573.35573.75522.25420000634.2564*3.65623.55613.85563.05Page 126

# Huntingdon Life Sciences 00

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Summary of FOB and Motor Activity Analyses Table 7

# Analysis of Data from Clean Air Act, Section 211,

Huntingdon Life Sciences Study 00-6130

Under a Contract with API

This report details the statistical analysis of data from a series of behavioral observations using rats

from Huntingdon Life Sciences Study 00-6130. There were 10 animals of each sex in each of four

exposure groups (control, low, medium, and high exposure).

The data were mailed to ExxonMobil Biomedical Sciences, Inc., by Huntingdon Life Sciences

[Huntingdon Life Sciences, Inc., Mettlers Road, East Millstone, NJ 08875] on a data CD. The

data were in Microsoft Excel® format and converted to a Statistical Analysis System (SAS) data

base using a SAS program. The statistical analyses were done in SAS® version 8.02. The final

report, computer output, and original data CD will be sent to Huntingdon Life Sciences, Inc. A

copy of the final report will be sent to API [American Petroleum Institute, 1220 L St., NW,

Washington, DC 20005-4070].

The behavioral observations were the Functional Observational Battery (FOB) and motor activity

observations. The FOB consisted of 30 observations grouped in five areas (home cage, handling,

open field, reflex, and other). The areas and variables are listed in Table I below. The FOB and

motor activity data were collected on all animals prior to the start of the study and at 4th and 8th and

13th weeks.

Some of the FOB variables are measured on a continuous scale and some are nominal or count

scale. The continuous data are printed in italics in Table I. 

# The statistical analysis of the

continuous FOB variables was by a mixed model analysis of covariance with a first order

autoregressive error structure on the time points [Galecki, 1994, Pinheiro and Bates, 2002]. The

pretest response was used as the covariate. The residuals from the model were tested for normality

by the Shapiro- Wilk test. Those variables that did not exhibit normally distributed residuals at the

0.01 level of significance were converted to a function of its percentile rank, then transformed by

the inverse normal distribution (the transformation is known as Blom's transformation where the

rank order is replaced by [rank-0. 375]/[n+0.25]), and reanalyzed [Blom, 1958].

The nominal and count data of the FOB were analyzed by a cumulative logit repeated measures

analysis with contrast terms to test for differences between control and exposed groups [Agresti,

1989]. For the measures that are count data (such as number ofrears or number of urinations), the

count responses may be grouped into several data ranges to allow sufficient data in the cells. The

statistical model partitions the variation of the response variable among the variables sex, exposure

group, week number and their interactions.

The motor activity experiment measured the number of beam breaks in an activity box during a

five-minute interval. During a single session the experiment ran for 12 consecutive intervals (1

hour total). Data were analyzed by a mixed model analysis of covariance with an unstructured

error relationship among the five-minute periods, and a first order autoregressive error structure

on weeks [Galecki, 1994, Pinheiro and Bates, 2002]. The pretest response was used as the

covariate. The residuals from the model were tested for normality by the Kolmogorov-Smimov

1 

# Page 128

# test. Those variables that did not exhibit normally distributed residuals at the 0.01 level of

significance were converted to a function of its percentile rank, then transformed by the inverse

normal distribution (the transformation is known as Blom's transformation where the rank order

is replaced by [rank-0. 375]/[n+0.25]), and reanalyzed [Blom, 1958].

RESULTS: Table I

Functional Observational Battery Areas and Measures

(italics indicate a measure analyzed as a continuous variable)



# Home Cage Evaluations

Posture

Vocalization

Palpebral closure



# Handling Evaluations

Ease of removal

Ease of handling

Chromodacryorrhea

Lacrimation

Salivation

Altered fur appearance



# Open Field Evaluations

Gait and posture

Locomotion

Arousal level

Exophthalmia

Piloerection

Defecation count

Urination count

Fasciculation

Tremors

Convulsions 

# Reflex Assessments

Response to visual stimuli

Response to auditory stimuli

Pain perception

Pupillary response

Sensitivity to light touch

Proprioception



# Other

Forelimb grip strength

Hindlimb grip strength

Air righting ability



# Landingfoot splay

Body weight

FOB Continuous

None of the four continuous measures indicated statistically significant exposure related

differences. The residuals from the forelimb grip strength, foot splay, and Blom transformed

hindlimb grip strength were normally distributed. The residuals from body weight (both observed

and Blom transformed data) and hindlimb grip strength (observed data) were not normally

distributed. Because the significance levels for the observed and Blom transformed body weight

were not near the 0.05 level of significance the deviation from normality was not considered crucial

2 

# Page 129

# in these analyses.

FOB Discrete

For the 26 measures on a nominal or count scale only 6 measures showed any variation in response:

posture, ease of removal, ease of handling, defecation count, urination count, and sensitivity to

light touch. For these 6 measures none indicated statistically significant exposure related

differences. For the remaining 20 variables all animals, or at most four responses (less than 1.5%

of the observations), showed the same response in all groups at all time points and were not

analyzed because of the lack of meaningful variation in response.

Motor Activity

The residuals from the motor activity data analysis and those from the Blom transformed data were

not normally distributed as is required for the statistical analysis. A possible explanation for the

non-normal distribution of the residuals is that approximately 27% of the observations are zero. It 

# is

difficult to have normally distributed residuals when a large percentage of the data have the same

value. However, inspection of the residuals shows a reasonably symmetric distribution with

slightly heavier tails than a normal distribution. Overall, these departures of the residuals from

normality are not thought to effect the analysis conclusions.

The results of the analyses indicated statistically significant differences in responses in exposure

levels for both the original data and the Blom transformed data (p<0.002).

Contrast analysis of the Blom transformed data showed that response from low and medium

exposure groups were significantly different from the control group. Least squares means of the

mean number of beam breaks across the three exposure groups are shown in Table II. The least

squares means are the means accounting for initial activity (pre-test), sex, study week, replicates,

and their interactions. There is no linear relationship between exposure levels and response, or any

consistent change in activity related to exposure. The attached data plots do not suggest a

consistent exposure related response, but show an increase in activity in the female control group

and decrease in activity in the female mid exposure group only during study week 8 (there was not a

statistically significant time by exposure effect).

Table II.

M ean 

# B B ak In 

# 1 earn re s ner terva

Exposure Least Squares Mean

Groun (Breaks/interval)

control 56.53

low 46.23**

medium 42.87**

high 52.18

**different from control p<O.Ol

3 

# Page 130

# CONCLUSION:

Based on these findings, administration of the test substance at the exposures tested is not

associated with any change in counts of the 26 nominal FOB measures, or the 4 continuous FOB

measures, or with consistent changes in motor activity.

# ~~__L/ Mark J. Nicolich,

# ~'**~~-""** OC'-



# Statistician

ExxonMobil Biomedical Sciences, Inc

1545 Route 22 East

PO Box 971

Annandale, NJ 08801-0971

# ~--~**~~**

George

# Bukh~r, Ph.D.

Consultant

References:

BLOM'S TRANSFORMATION 

# r/?fftr

July 24, 2009

Blom, G., Statistical Estimates and Transformed Beta Variables, J. Wiley and Sons, NY, 1958.

CUMULATIVE LOGIT

Agresti, Alan, "A survey of models for repeated ordered categorical response data", Statistics in

Medicine, vol8, 

# 1989, pg 1209-1224.

KOLMOGOROV -SMIRNOV TEST

Stephens, M.A., 'EDF statistics for goodness of fit and some comparisons', Journal ofthe American

Statistical Association, vol69, 1974, pg 730-737.

MIXED MODEL ANALYSES

Galecki, A.T., "General class of covariance structures for two or more repeated factors in

longitudinal data analysis," Communications in Statistics-Theory and Methods, 23(11), 1994 pg

3105 -3119.

Pinheiro, J.C. and Bates, D.M., Mixed-Effects Models in S and S-PLUS, Springer-Verlag, NY,

2002.

SAS

SAS/STAT, V8.02, copyright© 1999-2001, SAS Institute, Inc., Cary, NC.



# 4 

# Page 131

# SHAPIRO- WILK TEST

Shapiro, S.S. and Wilk, M.B., "An analysis of variance test for normality (complete samples),"

Biometrika, 52, 

# 1965, pg 

# 591-611.

5 

# Page 132225200nsc:175Q)....r:::150-..fQ)\c.125tnns1ooQ)....m1sr:::nsQ)50::?!25Pretest\\...\-.-\I••\\\1',\:·"'.\,:·.\I·.\ . ...,....,.I\··'=-,·.'-/\,·.\\·A(,Y,\o\MotorActivitvMeansStudy61!0Sex=FemaleWeek4WeekS\.\\\....:.f-i.r.\1:I·\1:I•\I'·I'\1··.I•I•\-·............'·.\'....'\.."'.·..0'-'-..i'·..._,..;><:.___../.......Week131 2 3 4 s 6 7 8 9a11 2 3 4 s 6 7 8 9a11 2 3 4 s 6 7 8 9a11 2 3 4 s 6 7 8 9a1TimePeriodDoseGroup-1---·2--3......4Page 133225-2001cuc:175(I)s::::150-'-8.125tJ)100(I)'-m75s::::cu(I)50::iPretest\_\.\IIIIIIIIIIIII1 2 3 4 5 6 7 8 9 1 1 101 2IIIIIMotorActivitvMeansStudySex=MaleWeek4WeekSI•'I:I:I'\I'\\·.I\'I'I',I\',I:I:y,\:),v·1•,I...\\\\\-·./,....\'''\.'·IIIIIIIIIIIIIIIIIIIIIIII1 2 3 4 5 6 7 8 9!1 2 3 4 5 6 7 8 9 1 1 101 2TimePeriodDoseGroup--1---·2--3......4Week13III1-IIIIIIIII1 2 3 4 5 6 7 8 9!Page 134MALEExposureWeekGroup1 20cmean116.20 143.60sd85.3037.27n1010Lmean119.80172.20sd89.1446.06n1010Mmean136.70140.10sd67.3079.16n1010Hmean120.90142.60sd92.6868.96n10104cmean193.60167.40sd32.1246.75n1010Lmean186.00144.90sd39.8948.78n1010Mmean158.00168.40sd92.7531.25n1010Hmean167.00179.40sd72.8132.98n10108cmean159.10121.70sd28.9265.69n10108 Lmean150.80119.90Study00-6130Mean,sd,andSampleSizeforBeamBreaksperIntervalBySex,StudyWeek,andExposureGroupInterval3 45 6 7149.30 127.90109.40105.5082.7048.7842.3438.3635.2152.131010101010136.70121.80107.80115.5087.0042.9759.7253.7652.8555.211010101010107.20119.10111.8073.0056.8047.9545.1244.13.53.9562.411010101010126.80 121.60102.6087.7051.6055.1952.4956.0352.5446.451010101010136.00139.80109.9078.4058.2042.3746.4162.4957.0256.331010101010111.3086.4085.4072.0046.2033.0069.3783.3569.2850.891010101010137.0091.2067.4084.4043.3038.5347.7053.6085.9065.181010101010127.40108.7072.9062.4043.7055.8650.5969.4053.9960.11101010101079.2071.7055.8041.9026.7055.2751.8459.6654.0248.32101010101055.7043.0025.5023.0019.008910111269.4057.9039.7021.0023.0055.5155.6944.8845.6942.26101010101062.7058.7016.0015.309.3048.5954.7326.6831.9022.08101010101043.6017.6010.304.503.2050.6438.0018.1913.197.74101010101055.6039.5021.805.901.5052.8268.8647.5512.622.46101010101056.6037.0036.9027.601.3061.9954.4455.6342.382.06101010101018.8022.8016.5021.3015.9030.8456.4043.1856.6344.04101010101031.7018.4013.602.403.9057.6829.9934.564.068.27101010101021.4026.507.1010.800.8037.0941.3011.4218.691.40101010101015.301.2011.5020.5019.9023.092.8222.3154.3944.7610101010101.207.6010.9020.9020.70Page 135ExposureIntervalWeekGroup1 23456 7 89101112sd54.2654.5940.9438.2141.0945.5821.961.4814.1815.3139.6140.82n101010101010101010101010Mmean163.0089.5058.5035.8040.0027.209.402.804.803.201.802.80sd51.2577.9752.2942.2152.0543.4814.185.098.756.863.462.49n101010101010101010101010Hmean155.30107.8083.5056.8044.0037.6040.7047.8034.2017.9018.1011.70sd60.7445.8442.0449.9642.6248.1343.8075.8156.9134.8341.8720.27n10101010101010101010101013cmean182.30133.10108.2053.4045.9037.5011.4017.5025.4037.3033.609.60sd56.8644.1031.5540.3631.4049.4910.7333.0449.1544.0943.0116.83n101010101010101010101010Lmean195.33119.8978.4445.4425.56 23.7833.7825.2225.0010.564.7818.33sd29.1730.5960.7847.5520.9927.0159.1438.9147.8628.326.6746.85n 9 9 9 9 9 99 9 9 9 9 9Mmean162.30 114.5071.8038.2025.7032.2016.1025.107.8020.1020.906.30sd61.0257.6349.1349.3639.0738.8428.4641.7213.3444.9137.807.85n101010101010101010101010Hmean173.50102.7072.9054.2048.2033.6010.5020.4032.7012.704.801.20sd41.9322.8339.8349.4059.4843.4715.3133.7346.0230.4210.282.10n101010101010101010101010Page 136FEMALEExposureIntervalWeekGroup1 2 3 45 6 7 8 91011120cmean119.10112.6099.8089.6066.4070.7043.6037.2025.9033.8017.50 13.90sd39.8226.6835.1853.1249.1655.5944.8550.0433.3957.6438.8730.05n101010101010101010101010Lmean140.10119.7076.1056.3070.0072.20 69.7067.3048.7053.8041.9023.40sd30.5835.4549.2145.6758.9961.28 64.5756.8143.9254.2560.6331.85n101010101010101010101010Mmean128.50 116.9079.8042.6041.0061.1054.1042.7049.1023.9016.30 12.60sd40.8840.9839.19 34.3756.7959.6964.3846.3360.3344.1733.5726.31n101010101010101010101010Hmean122.90118.4082.9080.2084.0068.50 65.2049.0050.5064.8044.6025.80sd52.8232.2740.8437.5840.9035.1546.8845.9153.3070.0243.6429.78n1010101010101010101010104cmean125.80116.0071.70 74.7055.7048.8043.8039.3020.706.304.400.50sd32.78 35.7434.5557.2449.2542.3151.1350.1927.0715.839.291.27n101010101010101010101010Lmean136.10121.60100.0080.1060.0050.1014.1011.009.203.905.402.10sd25.7717.8138.9649.2549.3064.2128.7322.3025.9710.6916.734.20n101010101010101010101010Mmean135.90119.9078.9062.1024.2032.6027.1016.401.500.300.702.60sd24.5853.9553.42 56.0436.8632.2247.2133.192.460.671.648.22n101010101010101010101010Hmean127.30120.00113.4063.6062.7061.8054.6039.6048.10 45.6023.6018.30sd50.5040.1429.6645.4944.0655.4355.1145.6054.3761.1839.5732.16n1010101010101010101010108cmean133.90112.3088.2072.2052.5048.8024.5014.1012.608.406.009.20sd38.2346.2352.2041.2242.2448.0936.9222.9724.5325.5212.6624.96n101010101010101010101010Lmean123.5073.5072.6029.9030.3017.9012.1019.308.70 8.704.904.40sd31.8736.9029.6034.0941.0532.0726.3143.1824.2027.519.8911.88n101010101010101010101010Page 137ExposureIntervalWeekGroup1 2 3 45 6 7 891011128Mmean123.9067.2046.709.103.700.100.001.3010.509.102.305.30sd42.3730.8752.8016.417.590.320.001.8328.0214.113.9514.74n101010101010101010101010Hmean136.30104.3075.3040.2025.8029.9019.7012.108.606.007.003.60sd45.98 45.7751.0338.6135.2539.2733.3033.4127.2018.62 12.178.59n10101010101010101010101013cmean137.50103.5089.5065.1038.6022.6014.5014.0011.9018.3015.104.10sd42.0638.7841.9247.9046.3929.3522.9227.4314.6736.0322.208.36n101010101010101010101010Lmean140.0068.3053.1023.8014.307.2023.9023.0023.4034.0022.507.20sd30.9417.6141.9627.1335.1313.2851.4542.1145.7752.7339.8320.02n101010101010101010101010Mmean128.8087.4053.0048.6015.6023.4013.5011.507.7014.1015.804.00sd40.8737.5452.0454.7019.6046.7131.7623.7011.4228.7141.278.63n101010101010101010101010Hmean148.1096.2070.8043.3042.0019.704.409.6023.302.401.702.70sd35.19 35.5552.4049.5232.0033.437.4720.0551.065.934.694.45n10Hl101010-10.....101010----101010Page 138

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Males Mean Motor Activity Values a



# Pretest Table 8

a 

# Number of beam breaks per interval Interval

1 2 3 4 5 6 7 8 9 10 11 12

Group I

- 0 mg/m 3



# Mean 116.2 143.6 149.3 127.9 109.4 105.5 82.7 69.4 57.9 39.7 21.0 23.0

SD 85.30 37.27 48.78 42.34 38.36 35.21 52.13 55.51 55.69 44.88 45.69 42.26

N 10 10 10 10 10 10 10 10 10 10 10 10

Group II

- 2000 mg/m 3



# Mean 119.8 172.2 136.7 121.8 107.8 115.5 87.0 62.7 58.7 16.0 15.3 9.3

SD 89.14 46.06 42.97 59.72 53.76 52.85 55.21 48.59 54.73 26.68 31.90 22.08

N 10 10 10 10 10 10 10 10 10 10 10 10

Group III

- 10000 mg/m 3



# Mean 136.7 140.1 107.2 119.1 111.8 73.0 56.8 43.6 17.6 10.3 4.5 3.2

SD 67.30 79.16 47.95 45.12 44.13 53.95 62.41 50.64 38.00 18.19 13.19 7.74

N 10 10 10 10 10 10 10 10 10 10 10 10

Group IV

- 20000 mg/m 3



# Mean 120.9 142.6 126.8 121.6 102.6 87.7 51.6 55.6 39.5 21.8 5.9 1.5

SD 92.68 68.96 55.19 52.49 56.03 52.54 46.45 52.82 68.86 47.55 12.62 2.46

N 10 10 10 10 10 10 10 10 10 10 10 10

# Huntingdon Life Sciences 00

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Females Mean Motor Activity Values a



# Pretest Table 8

a 

# Number of beam breaks per interval Interval

1 2 3 4 5 6 7 8 9 10 11 12

Group I

- 0 mg/m 3



# Mean 119.1 112.6 99.8 89.6 66.4 70.7 43.6 37.2 25.9 33.8 17.5 13.9

SD 39.82 26.68 35.18 53.12 49.16 55.59 44.85 50.04 33.39 57.64 38.87 30.05

N 10 10 10 10 10 10 10 10 10 10 10 10

Group II

- 2000 mg/m 3



# Mean 140.1 119.7 76.1 56.3 70.0 72.2 69.7 67.3 48.7 53.8 41.9 23.4

SD 30.58 35.45 49.21 45.67 58.99 61.28 64.57 56.81 43.92 54.25 60.63 31.85

N 10 10 10 10 10 10 10 10 10 10 10 10

Group III

- 10000 mg/m 3



# Mean 128.5 116.9 79.8 42.6 41.0 61.1 54.1 42.7 49.1 23.9 16.3 12.6

SD 40.88 40.98 39.19 34.37 56.79 59.69 64.38 46.33 60.33 44.17 33.57 26.31

N 10 10 10 10 10 10 10 10 10 10 10 10

Group IV

- 20000 mg/m 3



# Mean 122.9 118.4 82.9 80.2 84.0 68.5 65.2 49.0 50.5 64.8 44.6 2 5.8

SD 52.82 32.27 40.84 37.58 40.90 35.15 46.88 45.91 53.30 70.02 43.64 29.78

N 10 10 10 10 10 10 10 10 10 10 10 10

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# 4 th 

# week of exposure Table 8

a 

# Number of beam breaks per interval Interval

1 2 3 4 5 6 7 8 9 10 11 12

Group I

- 0 mg/m 3



# Mean 193.6 167.4 136.0 139.8 109.9 78.4 58.2 56.6 37.0 3 6.9 27.6 1.3

SD 32.12 46.75 42.37 46.41 62.49 57.02 56.33 61.99 54.44 55.63 42.38 2.06

N 10 10 10 10 10 10 10 10 10 10 10 10

Group II

- 2000 mg/m 3



# Mean 186.0 144.9 111.3 86.4 85.4 72.0 46.2 18.8 22.8 16.5 21.3 15.9

SD 39.89 48.78 33.00 69.37 83.35 69.28 50.89 30.84 56.40 43.18 56.63 44.04

N 10 10 10 10 10 10 10 10 10 10 10 10

Group III

- 10000 mg/m 3



# Mean 158.0 168.4 137.0 91.2 67.4 84.4 43.3 31.7 18.4 13.6 2.4 3.9

SD 92.75 31.25 38.53 47.70 53.60 85.90 65.18 57.68 29.99 34.56 4.06 8.27

N 10 10 10 10 10 10 10 10 10 10 10 10

Group IV

- 20000 mg/m 3



# Mean 167.0 179.4 127.4 108.7 72.9 62.4 43.7 21.4 26.5 7.1 10.8 0.8

SD 72.81 32.98 55.86 50.59 69.40 53.99 60.11 37.09 41.30 11.42 1 8.69 1.40

N 10 10 10 10 10 10 10 10 10 10 10 10

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# 4 th 

# week of exposure Table 8

a 

# Number of beam breaks per interval Interval

1 2 3 4 5 6 7 8 9 10 11 12

Group I

- 0 mg/m 3



# Mean 125.8 116.0 71.7 74.7 55.7 48.8 43.8 39.3 20.7 6.3 4.4 0.5

SD 32.78 35.74 34.55 57.24 49.25 42.31 51.13 50.19 27.07 15.83 9.29 1.27

N 10 10 10 10 10 10 10 10 10 10 10 10

Group II

- 2000 mg/m 3



# Mean 133.9 120.7 95.4 79.4 56.9 53.7 17.3 12.1 10.1 3.9 5.4 2.1

SD 25.00 18.61 37.31 48.27 45.89 66.66 30.92 22.62 25.77 10.69 16.73 4.20

N 10 10 10 10 10 10 10 10 10 10 10 10

Group III

- 10000 mg/m 3



# Mean 135.9 119.9 78.9 62.1 24.2 32.6 27.1 16.4 1.5 0.3 0.7 2.6

SD 24.58 53.95 53.42 56.04 36.86 32.22 47.21 33.19 2.46 0.67 1.64 8.22

N 10 10 10 10 10 10 10 10 10 10 10 10

Gr oup IV

- 20000 mg/m 3



# Mean 127.3 120.0 113.4 63.6 62.7 61.8 54.6 39.6 48.1 45.6 23.6 18.3

SD 50.50 40.14 29.66 45.49 44.06 55.43 55.11 45.60 54.37 61.18 39.57 32.16

N 10 10 10 10 10 10 10 10 10 10 10 10

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# 8 th 

# week of exposure Table 8

a 

# Number of beam breaks per interval Interval

1 2 3 4 5 6 7 8 9 10 11 12

Group I

- 0 mg/m 3



# Mean 159.1 121.7 79.2 71.7 55.8 41.9 26.7 15.3 1.2 11.5 20.5 19.9

SD 28.92 65.69 55.27 51.84 59.66 54.02 48.32 23.09 2.82 22.31 54.39 44.76

N 10 10 10 10 10 10 10 10 10 10 10 10

Group II

- 2000 mg/m 3



# Mean 150.8 119.9 55.7 43.0 25.5 23.0 19.0 1.2 7.6 10.9 20.9 20.7

SD 54.26 54.59 40.94 38.21 41.09 45.58 21.96 1.48 14.18 15.31 39.61 40.82

N 10 10 10 10 10 10 10 10 10 10 10 10

Group III

- 10000 mg/m 3



# Mean 167.0 99.2 57 .3 33.3 55.8 32.8 13.3 2.9 7.6 1.4 1.8 2.8

SD 56.84 89.93 51.85 38.35 62.50 43.48 16.48 5.04 11.19 2.07 3.46 2.49

N 10 10 10 10 10 10 10 10 10 10 10 10

Group IV

- 20000 mg/m 3



# Mean 155.3 107.8 83.5 56.8 44.0 37.6 40.7 47.8 34 .2 17.9 18.1 11.7

SD 60.74 45.84 42.04 49.96 42.62 48.13 43.80 75.81 56.91 34.83 41.87 20.27

N 10 10 10 10 10 10 10 10 10 10 10 10

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# 8 th 

# week of exposure Table 8

a 

# Number of beam breaks per interval Interval

1 2 3 4 5 6 7 8 9 10 11 12

Group I

- 0 mg/m 3



# Mean 133.9 112.3 88.2 72.2 52.5 48.8 24.5 14 .1 12.6 8.4 6.0 9.2

SD 38.23 46.23 52.20 41.22 42.24 48.09 36.92 22.97 24.53 25.52 12.66 24.96

N 10 10 10 10 10 10 10 10 10 10 10 10

Group II

- 2000 mg/m 3



# Mean 123.5 73.5 72.6 29.9 30.3 17.9 12.1 19.3 8.7 8.7 4.9 4.4

SD 31. 87 36.90 29.60 34.09 41.05 32.07 26.31 43.18 24.20 27.51 9.89 11.88

N 10 10 10 10 10 10 10 10 10 10 10 10

Group III

- 10000 mg/m 3



# Mean 123.9 67.2 46.7 9.1 3.7 0.1 0.0 1.3 10.5 9.1 2.3 5.3

SD 42.37 30.87 52.80 16.41 7.59 0.32 0.00 1.83 2 8.02 14.11 3.95 14.74

N 10 10 10 10 10 10 10 10 10 10 10 10

Group IV

- 20000 mg/m 3



# Mean 136.3 104.3 75.3 40.2 25.8 29.9 19.7 12.1 8.6 6.0 7.0 3.6

SD 45.98 45.77 51.03 38.61 35.25 39.27 33.30 33.41 27.20 18.62 12.17 8.59

N 1 0 10 10 10 10 10 10 10 10 10 10 10

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# 13 th 

# week of exposure Table 8

a 

# Number of beam breaks per interval

b 

# Animal 2070 sacrificed for humane reason

. Interval

1 2 3 4 5 6 7 8 9 10 11 12

Group I

- 0 mg/m 3



# Mean 182.3 133.1 108.2 53.4 45.9 37.5 11.4 17.5 25.4 37.3 33.6 9.6

SD 56.86 44.10 31.55 40.36 31.40 49.49 10.73 33.04 49.15 44.09 43.01 16.83

N 10 10 10 10 10 10 10 10 10 10 10 10

Group II

- 2000 mg/m 3



# Mean 195.3 119.9 78.4 45.4 25.6 23.8 33.8 25.2 25.0 10.6 4.8 18.3

SD 29.17 30.59 60.78 47.55 20.99 27.01 59.14 38.91 47.86 28.32 6.67 46.85

N 9 b 

# 9 9 9 9 9 9 9 9 9 9 9

Group III

- 10000 mg/m 3



# Mean 162.3 114.5 71.8 38.2 25.7 32.2 16.1 25.1 7.8 20.1 20.9 6.3

SD 61.02 57.63 49.13 49.36 39.07 38.84 28.46 41.72 13.34 44.91 37.80 7.85

N 10 10 10 10 10 10 10 10 10 10 10 10

Group IV

- 20000 mg/m 3



# Mean 173.5 102.7 72.9 54.2 48.2 33.6 10.5 20.4 32.7 12.7 4.8 1.2

SD 41.93 22.83 39.83 49.40 59.48 43.47 15.31 33.73 46.02 30.42 10.28 2.10

N 10 10 10 10 10 10 10 10 10 10 10 10

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# 13 th 

# week of exposure Table 8

Interval

1 2 3 4 5 6 7 8 9 10 11 12

Group I

- 0 mg/m 3



# Mean 137.5 103.5 89.5 65.1 38.6 22.6 14.5 14.0 11.9 18.3 15.1 4.1

SD 42.06 38.78 41.92 47.90 46.39 29.35 22.92 27.43 14.67 36.03 22.20 8.36

N 10 10 10 10 10 10 10 10 10 10 10 10

Group II

- 2000 mg/m 3



# M ean 140.0 68.3 53.1 23.8 14.3 7.2 23.9 23.0 23.4 34.0 22.5 7.2

SD 30.94 17.61 41.96 27.13 35.13 13.28 51.45 42.11 45.77 52.73 39.83 20.02

N 10 10 10 10 10 10 10 10 10 10 10 10

Group III

- 10000 mg/m 3



# Mean 128.8 87.4 53.0 48.6 15.6 23.4 13.5 11.5 7.7 14.1 15.8 4.0

SD 40.87 37.54 52.04 54.70 19.60 46.71 31.76 23.70 11.42 28.71 41.27 8.63

N 10 10 10 10 10 10 10 10 10 10 10 10

Group IV

- 20000 mg/m 3



# Mean 148.1 96.2 70.8 43.3 42.0 19.7 4.4 9.6 23.3 2.4 1.7 2.7

SD 35.19 35.55 52.40 49.52 32.00 33.43 7.47 20.05 51.06 5.93 4.69 4.45

N 10 10 10 10 10 10 10 10 10 10 10 10

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Pretest Table 9

*Significantly different from control mean; p ≤ 0.05.

**Significantly diff erent from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean. Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Body Weight (grams) Mean 256.8 256.5 254.3 260.8

S.D. 17.0 23.0 19.3 23.2

N 10 10 10 10

Forelimb Grip Strength (grams of force)

Mean 878.3 950.0 878.0 967.3

S.D. 116.8 128.0 84.9 105.7

N 10 10 10 10

Hindlimb Grip Strength (grams of force)

Mean 585.0 593.8 548.0 612.5

S.D. 114.9 143.4 100.4 134.5

N 10 10 10 10

Landing Foot Splay ( centimeters)

Mean 6.2 6.2 6.3 6.9

S.D. 1.4 1.4 1.0 1.0

N 10 10 10 10

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Pretest Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals

Examined 10 10 10 10

Home Cage Eval uations

Posture Sitting or Standing Normally 9 10 10 10

Alert, Oriented Toward Observer 1 0 0 0

Asleep 0 0 0 0

Lying on Side 0 0 0 0

Flattened 0 0 0 0

Crouched 0 0 0 0

Vocalizations Not Present 10 10 10 10

Present 0 0 0 0

Palpebral Closure Eyelids Open 10 10 10 10

Eyelids Completely Closed 0 0 0 0

Eyelids Slightly Drooping 0 0 0 0

Eyelids Half Closed 0 0 0 0

Handling Evaluations

Ease of Removal Very Easy 10 9 10 8

Easy 0 1 0 2

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Ease of Handling Very easy; totally limp 2 3 2 3

Normal 8 7 8 7

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Chromodacryorrhea Not Present 10 10 10 10

Present 0 0 0 0

Lacrimation No lacrimation 10 10 10 10

Some lacrimation 0 0 0 0

Extreme lacrimation 0 0 0 0

Salivation Not Present 10 10 10 10

Slight 0 0 0 0

Moderate 0 0 0 0

Extreme 0 0 0 0

Coat Normal 10 10 10 10

Slightly Soiled 0 0 0 0

Moderately Soiled 0 0 0 0

Extremely Soiled 0 0 0 0

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Pretest Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals

Examined 10 10 10 10

Open Field Evaluations

Gait Normal 10 10 10 10

Ata xia 0 0 0 0

Body Drags 0 0 0 0

Limbs splayed or dragging 0 0 0 0

Hunched posture 0 0 0 0

Locomotion Normal 10 10 10 10

Increased movement 0 0 0 0

Decreased movement 0 0 0 0

None 0 0 0 0

Arousal Normal 10 10 10 10

Low, Slight S tupor 0 0 0 0

Very Low, Stupor 0 0 0 0

High, Slight Excitement, Tense 0 0 0 0

Very High, Hyper

- Alert 0 0 0 0

Exophthalmia Not Present 10 10 10 10

Present 0 0 0 0

Piloerection Not Present 10 10 10 10

Present 0 0 0 0

Feces N umber of pellets 9 3 6 10

Unformed Stool 0 0 0 0

Urine Number of Pools 3 4 17 8

Polyuria 0 0 0 0

Fasciculation Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Tremors Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Convulsions Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

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Pretest Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals

Examined 10 10 10 10

Reflex Assessments

V isual Approach Normal, slowly approaches 10 10 10 10

No Reaction 0 0 0 0

Freezes or pulls away slightly 0 0 0 0

Jumps or Turns Abruptly to Avoid, or

attacks and/or bites 0 0 0 0

Audition Assessment Normal 10 10 10 10

No Reaction 0 0 0 0

E xaggerated, Jumps, Flips, Bite 0 0 0 0

Pain Perception Normal 10 10 10 10

No Reaction 0 0 0 0

Increased Pain 0 0 0 0

Pupil Response Pupil Constricts Normally 10 10 10 10

Pupil Size does not Change 0 0 0 0

Miosis 0 0 0 0

Mydriasis 0 0 0 0

Proprioception Returns leg to original position 10 10 10 10

Returns leg only partially to original

position 0 0 0 0

No response 0 0 0 0

Pinna Reflex Ear flattens against head 0 0 0 0

Animals shakes head 10 10 10 10

No respon se 0 0 0 0

Air Righting Reflex Lands on All Four Feet (Normal) 10 10 10 10

Slightly Uncoordinated 0 0 0 0

Lands on Side 0 0 0 0

Lands on Back 0 0 0 0

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Pretest Table 9

*Significantly different from control mean; p ≤ 0.05.

**Significantly different from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean. Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Body Weight (grams) Mean 197.0 194.6 195.5 195.1

S.D. 10.2 8.5 13.3 11.1

N 10 10 10 10

Forelimb Grip Strength (grams of force)

Mean 884.5 920.3 898.0 894.5

S.D. 108.8 136.1 84.7 129.8

N 10 10 10 10

Hindlimb Grip Strength (grams of force)

Mean 661.3 596.5 583.0 625.3

S.D. 94.5 73.7 108.3 88.5

N 10 10 10 10

Landing Foot Splay (centimeters)

Mean 4.9 5.2 5.1 5.3

S.D. 0.7 0.6 1.2 1.2

N 10 10 10 10

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Pretest Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Home Cage Evaluations

Posture Sitting or Standing Normally 9 8 9 10

Alert, Oriented Toward Observer 1 2 1 0

Asleep 0 0 0 0

Lying on Side 0 0 0 0

Flattened 0 0 0 0

Crouched 0 0 0 0

Vocalizations Not Present 10 10 10 10

Present 0 0 0 0

Palpebral Closure Eyelids Open 10 10 10 10

Eyelids Completely Closed 0 0 0 0

Eyelids Slight ly Drooping 0 0 0 0

Eyelids Half Closed 0 0 0 0

Handling Evaluations

Ease of Removal Very Easy 10 10 10 10

Easy 0 0 0 0

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Ease of Handling Very easy; totally limp 3 2 3 2

Normal 7 8 7 8

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Chromodacryorrhea Not Present 10 10 10 10

Present 0 0 0 0

Lacrimation No lacrimation 10 10 10 10

Some lacrimation 0 0 0 0

Extreme lacrimatio n 0 0 0 0

Salivation Not Present 10 10 10 10

Slight 0 0 0 0

Moderate 0 0 0 0

Extreme 0 0 0 0

Coat Normal 10 10 10 10

Slightly Soiled 0 0 0 0

Moderately Soiled 0 0 0 0

Extremely Soiled 0 0 0 0

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Pretest Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Open Field Evaluations

Gait Normal 10 10 10 10

Ataxia 0 0 0 0

Body Drags 0 0 0 0

Limbs splayed or dragging 0 0 0 0

Hunched posture 0 0 0 0

Locomotion Normal 10 10 10 10

Increased movement 0 0 0 0

Decreased movement 0 0 0 0

None 0 0 0 0

Arousal Normal 10 10 10 10

Low, Slight Stupor 0 0 0 0

Very Low, Stupor 0 0 0 0

High, Slight Excitement, Tense 0 0 0 0

Very High, Hyper

- Alert 0 0 0 0

Exophthalmia Not Present 10 10 10 10

Present 0 0 0 0

Piloerection Not Present 10 10 10 10

Present 0 0 0 0

Feces Number of pellets 1 0 0 1

Unformed Stool 0 0 0 0

Urine Number of Pools 3 0 7 2

Poly uria 0 0 0 0

Fasciculation Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Tremors Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Convulsions Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

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Pretest Table 9

Gro up I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Reflex Assessments

Visual Approach Normal, slowly approaches 10 10 10 10

No Reaction 0 0 0 0

Freezes or pulls aw ay slightly 0 0 0 0

Jumps or Turns Abruptly to Avoid,

or attacks and/or bites 0 0 0 0

Audition Assessment Normal 10 10 10 10

No Reaction 0 0 0 0

Exaggerated, Jumps, Flips, Bite 0 0 0 0

Pain Perception Normal 10 10 10 10

No Reaction 0 0 0 0

Increased Pain 0 0 0 0

Pupil Response Pupil Constricts Normally 10 10 10 10

Pupil Size does not Change 0 0 0 0

Miosis 0 0 0 0

Mydriasis 0 0 0 0

Proprioception Returns leg to original position 10 10 10 10

Returns leg only p artially to original

position 0 0 0 0

No response 0 0 0 0

Pinna Reflex Ear flattens against head 0 0 0 1

Animals shakes head 10 10 10 9

No response 0 0 0 0

Air Righting Reflex Lands on All Four Feet (Normal) 10 10 10 10

Slightly Unc oordinated 0 0 0 0

Lands on Side 0 0 0 0

Lands on Back 0 0 0 0

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# week of exposure Table 9

*Significantly different from control mean; p ≤ 0.05.

**Significantly different from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean. Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Body Weight (grams) Mean 397.1 396.3 410.1 405.5

S.D. 33.3 37.0 30.0 23.1

N 10 10 10 10

Forelimb Grip Strength (grams of force)

Mean 1270.3 1327.3 1328.0 1327.5

S.D. 139.0 139.1 218.0 137.5

N 10 10 10 10

Hindlimb Grip Strength (grams of force)

Mean 834.5 780.8 848.8 883.8

S.D. 126.7 1 81.3 273.2 206.9

N 10 10 10 10

Landing Foot Splay (centimeters)

Mean 7.1 6.5 6.4 7.0

S.D. 1.7 0.9 1.4 1.8

N 10 10 10 10

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4 th 

# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Home Cage Evaluations

Posture Sitting or Standing Normally 9 9 8 9

Alert, Oriented Toward Observer 1 1 2 1

Asleep 0 0 0 0

Lying on Side 0 0 0 0

Flattened 0 0 0 0

Crouched 0 0 0 0

Vocalizations Not Present 10 10 10 10

Present 0 0 0 0

Palpebral Closure Eyelids Open 10 10 10 10

Eyelids Completely Closed 0 0 0 0

Eyelids Slightly Drooping 0 0 0 0

Eyelids Half Closed 0 0 0 0

Handling Evaluations

Ease of Removal Very Easy 10 10 7 10

Easy 0 0 3 0

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Ease of Handling Very easy; totally limp 0 0 0 0

Normal 10 10 10 10

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Chromodacryorrhea Not Present 10 10 10 10

Present 0 0 0 0

Lacrimation No lacrimation 10 10 10 10

Some lacrimation 0 0 0 0

Extreme lacrimation 0 0 0 0

Salivation Not Present 10 10 10 10

Slight 0 0 0 0

Moderate 0 0 0 0

Ex treme 0 0 0 0

Coat Normal 10 10 10 10

Slightly Soiled 0 0 0 0

Moderately Soiled 0 0 0 0

Extremely Soiled 0 0 0 0

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4 th 

# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Open Field Evaluations

Gait Normal 10 10 10 10

Ataxia 0 0 0 0

Body Drags 0 0 0 0

Limbs splayed or dragging 0 0 0 0

Hunched posture 0 0 0 0

Locomotion Normal 10 10 10 10

Increased movement 0 0 0 0

Decreased movement 0 0 0 0

None 0 0 0 0

Arousal Normal 10 10 10 10

Low, Slight Stupor 0 0 0 0

Very Low, Stupor 0 0 0 0

High, Slight Excitement, Tense 0 0 0 0

Very High, Hyper

- Alert 0 0 0 0

Exophthalmia Not Present 10 10 10 10

Present 0 0 0 0

Pi loerection Not Present 10 10 10 10

Present 0 0 0 0

Feces Number of pellets 10 5 25 14

Unformed Stool 0 0 0 0

Urine Number of Pools 12 18 9 8

Polyuria 0 0 0 0

Fasciculation Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Tremors Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Convulsions Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Numb er of Animals Examined 10 10 10 10

Reflex Assessments

Visual Approach Normal, slowly approaches 9 10 10 10

No Reaction 1 0 0 0

Freezes or pulls away slightly 0 0 0 0

Jumps or Turns Abruptly to Avoid, or

attacks and/or bites 0 0 0 0

Audition Assessment Normal 10 10 10 10

No Reaction 0 0 0 0

Exaggerated, Jumps, Flips, Bite 0 0 0 0

Pain Perception Normal 10 10 10 10

No Reaction 0 0 0 0

Increased Pain 0 0 0 0

Pupil Response Pupil Constricts Normally 10 10 10 1 0

Pupil Size does not Change 0 0 0 0

Miosis (bilateral) 0 0 0 0

Mydriasis 0 0 0 0

Proprioception Returns leg to original position 10 10 10 10

Returns leg only partially to original

position 0 0 0 0

No response 0 0 0 0

Pinna Reflex Ear flattens against head 0 1 0 0

Animals shakes head 10 9 10 10

No response 0 0 0 0

Air Righting Reflex Lands on All Four Feet (Normal) 10 10 10 10

Slightly Uncoordinated 0 0 0 0

Lands on Side 0 0 0 0

Lands on Back 0 0 0 0

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*Significantly different from control mean; p ≤ 0.05.

**Significantly different from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean. Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Body Weight (grams) Mean 271.5 262.3 254.4 253.3

S.D. 19.4 22.8 26.3 13.1

N 10 10 10 10

Forelimb Grip Strength (grams of force)

Mean 1179.0 117 1.5 1172.8 1184.0

S.D. 74.1 107.9 196.8 106.4

N 10 10 10 10

Hindlimb Grip Strength (grams of force)

Mean 787.3 767.0 783.8 712.0

S.D. 128.9 141.9 104.8 112.3

N 10 10 10 10

Landing Foot Splay (centimeters)

Mean 5.4 5.2 5.5 6.0

S.D. 0.8 0.8 1.2 1.2

N 10 10 10 10

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Home Cage Evaluations

Posture Sitting or Standing Normally 7 6 6 5

Alert, Oriented Toward Observer 2 3 4 5

Asleep 1 1 0 0

Lying on Side 0 0 0 0

Flattened 0 0 0 0

Crouched 0 0 0 0

Vocalizations Not Present 10 10 10 10

Present 0 0 0 0

Palpebral Closure Eyelids Open 9 9 10 10

Eyelids Completely Closed 1 1 0 0

Eyelids Slightly Drooping 0 0 0 0

Eyelids Half Closed 0 0 0 0

Handling Evaluations

Ease of Removal Very Easy 10 10 10 9

Easy 0 0 0 1

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Ease of Handling Normal 0 1 2 0

Moderately Difficult 10 9 8 10

Freezes 0 0 0 0

Difficult 0 0 0 0

Totally limp 0 0 0 0

Chromodacryorrhea Not Present 10 10 10 10

Present 0 0 0 0

Lacrimation No lacrimation 10 10 10 10

Some lacrimation 0 0 0 0

Extreme lacrimation 0 0 0 0

Salivation Not Present 10 10 10 10

Slight 0 0 0 0

Moderate 0 0 0 0

Extreme 0 0 0 0

Coat Normal 10 10 10 10

Slightly Soiled 0 0 0 0

Moderately Soiled 0 0 0 0

Extremely Soiled 0 0 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Open Field Evaluations

Gait Normal 10 10 10 10

Ataxia 0 0 0 0

Body Drags 0 0 0 0

Limbs splayed or dragging 0 0 0 0

Hunched posture 0 0 0 0

Locomotion Normal 10 10 10 10

Increased movement 0 0 0 0

Decreased movement 0 0 0 0

None 0 0 0 0

Arousal Normal 10 10 10 10

Low, Slight Stupor 0 0 0 0

Very Low, Stu por 0 0 0 0

High, Slight Excitement, Tense 0 0 0 0

Very High, Hyper

- Alert 0 0 0 0

Exophthalmia Not Present 10 10 10 10

Present 0 0 0 0

Piloerection Not Present 10 10 10 10

Present 0 0 0 0

Feces Number of pellets 4 0 1 0

Un formed Stool 0 0 0 0

Urine Number of Pools 2 0 3 0

Polyuria 0 0 0 0

Fasciculation Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Tremors Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Convulsions No t Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Reflex Assessments

Visual Approach Normal, slowly app roaches 10 10 10 10

No Reaction 0 0 0 0

Freezes or pulls away slightly 0 0 0 0

Jumps or Turns Abruptly to Avoid,

or attacks and/or bites 0 0 0 0

Audition Assessment Normal 10 10 10 10

No Reaction 0 0 0 0

Exaggerated, Jumps, Flips, Bite 0 0 0 0

Pain Perception Normal 10 10 10 10

No Reaction 0 0 0 0

Increased Pain 0 0 0 0

Pupil Response Pupil Constricts Normally 10 10 10 10

Pupil Size does not Change 0 0 0 0

Miosis (bilateral) 0 0 0 0

Mydriasis 0 0 0 0

Propr ioception Returns leg to original position 10 10 10 10

Returns leg only partially to

original position 0 0 0 0

No response 0 0 0 0

Pinna Reflex Ear flattens against head 1 0 1 0

Animals shakes head 9 10 9 10

No response 0 0 0 0

Air Righting Reflex Lands on All Four Feet (Normal) 10 10 10 10

Slightly Uncoordinated 0 0 0 0

Lands on Side 0 0 0 0

Lands on Back 0 0 0 0

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*Significantly different from control mean; p ≤ 0.05.

**Significantly different from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean. Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Body Weight (g rams) Mean 485.1 463.1 504.9 501.8

S.D. 46.7 85.2 35.1 44.5

N 10 10 10 10

Forelimb Grip Strength (grams of force)

Mean 1207.3 1175.0 1138.0 1266.0

S.D. 175.9 230.0 228.6 192.2

N 10 10 10 10

Hindlimb Grip Strength (grams of force)

Mean 921.5 803.8 838.5 859.0

S.D. 195.1 172.6 351.5 170.4

N 10 10 10 10

Landing Foot Splay (centimeters)

Mean 7.1 6.3 5.9 6.3

S.D. 1.7 1.0 1.4 0.8

N 10 10 10 10

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# week of exposure Table 9

Group I G roup II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals

Examined 10 10 10 10

Home Cage Evaluations

Posture Sitting or Standing Normally 8 8 8 5

Alert, Oriented Toward Observer 2 2 2 5

Asleep 0 0 0 0

Lying on Side 0 0 0 0

Flattened 0 0 0 0

Crouched 0 0 0 0

Vocalizations Not Present 10 10 10 10

Present 0 0 0 0

Palpebral Closure Eyelids Open 10 10 10 10

Eyelids Completely Closed 0 0 0 0

Eyelids Slightly Drooping 0 0 0 0

Eyelids Half Closed 0 0 0 0

Handling Evaluations

Ease of Removal Very Easy 7 8 10 8

Easy 3 2 0 2

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Ease of Handling Very easy; totally limp 0 0 0 0

Normal 10 10 10 10

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Chromodacryorrhea Not Present 10 9 10 10

Present 0 1 0 0

Lacrimation No lacrimation 10 9 10 10

Some lacrimation 0 1 0 0

Extreme lacrimation 0 0 0 0

Saliv ation Not Present 10 10 10 10

Slight 0 0 0 0

Moderate 0 0 0 0

Extreme 0 0 0 0

Coat Normal 10 10 10 10

Slightly Soiled 0 0 0 0

Moderately Soiled 0 0 0 0

Extremely Soiled 0 0 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (m g/m 3 

# ): 0 2000 10000 20000

Number of Animals

Examined 10 10 10 10

Open Field Evaluations

Gait Normal 10 9 10 10

Ataxia 0 0 0 0

Body Drags 0 0 0 0

Limbs splayed or dragging 0 0 0 0

Hunched posture 0 1 0 0

Locomotion Normal 10 9 10 10

Increased movement 0 0 0 0

Decreased movement 0 1 0 0

None 0 0 0 0

Arousal Normal 10 9 10 10

Low, Slight Stupor 0 1 0 0

Very Low, Stupor 0 0 0 0

High, Slight Excitement, Tense 0 0 0 0

Very High, Hyper

- Alert 0 0 0 0

Exophthalmia Not Present 10 10 10 10

Present 0 0 0 0

Piloerection Not Present 10 10 10 10

Present 0 0 0 0

Feces Number of pellets 9 11 14 12

Unformed Stool 0 0 0 0

Urine Number of Pools 8 9 13 6

Polyuria 0 0 0 0

F asciculation Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Tremors Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Convulsions Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

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# week of exposure Table 9

Group I Group II Group II I Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals

Examined 10 10 10 10

Reflex Assessments

Visual Approach Normal, slowly approaches 10 10 10 10

No Reaction 0 0 0 0

Freezes or pulls away slightly 0 0 0 0

Jumps or Turns Abruptly to Avoid, or

attacks and/or bites 0 0 0 0

Audition Assessment Normal 10 10 10 10

No Reaction 0 0 0 0

Exaggerated, Jumps, Flips, Bite 0 0 0 0

Pain Perception Normal 10 10 10 10

No Reaction 0 0 0 0

Increased Pa in 0 0 0 0

Pupil Response Pupil Constricts Normally 10 10 10 10

Pupil Size does not Change 0 0 0 0

Miosis 0 0 0 0

Mydriasis 0 0 0 0

Proprioception Returns leg to original position 10 10 10 10

Returns leg only partially to original

position 0 0 0 0

No response 0 0 0 0

Pinna Reflex Ear flattens against head 0 0 0 0

Animals shakes head 10 10 10 10

No response 0 0 0 0

Air Righting Reflex Lands on All Four Feet (Normal) 10 10 10 10

Slightly Uncoordinated 0 0 0 0

Lands on Side 0 0 0 0

Lands on Back 0 0 0 0

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*Significantly different from control mean; p ≤ 0.05.

**Significantly different from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean. Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Body Weight (grams) Mean 302.5 288.8 280.3 282.3

S.D. 29.3 26.5 33.7 18.9

N 10 10 10 10

Forelimb Grip Strength (grams of force)

Mean 1169.3 1029.8 1148.5 1018.0

S.D. 129.5 152.9 205.0 177.3

N 10 10 10 10

Hindlimb Grip Strength (grams of force)

Mean 774.8 775.0 803.5 741.3

S.D. 166.5 148.1 115.2 130.8

N 10 10 10 10

Landing Foot Splay (centimeters)

Mean 5.0 5.0 4.8 5.5

S.D. 0.4 0.8 0.9 0.9

N 10 10 10 10

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Anima ls Examined 10 10 10 10

Home Cage Evaluations

Posture Sitting or Standing Normally 4 5 6 5

Alert, Oriented Toward Observer 5 5 4 5

Asleep 1 0 0 0

Lying on Side 0 0 0 0

Flattened 0 0 0 0

Crouched 0 0 0 0

Vocalizations Not Pr esent 10 10 10 10

Present 0 0 0 0

Palpebral Closure Eyelids Open 9 10 10 10

Eyelids Completely Closed 1 0 0 0

Eyelids Slightly Drooping 0 0 0 0

Eyelids Half Closed 0 0 0 0

Handling Evaluations

Ease of Removal Very Easy 10 10 10 10

Easy 0 0 0 0

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Ease of Handling Very easy; totally limp 0 0 0 0

Normal 10 10 10 10

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Chromodacryo rrhea Not Present 10 10 10 10

Present 0 0 0 0

Lacrimation No lacrimation 10 10 10 10

Some lacrimation 0 0 0 0

Extreme lacrimation 0 0 0 0

Salivation Not Present 10 10 10 10

Slight 0 0 0 0

Moderate 0 0 0 0

Extreme 0 0 0 0

Coat Normal 10 10 10 10

Slightly Soiled 0 0 0 0

Moderately Soiled 0 0 0 0

Extremely Soiled 0 0 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Open Fiel d Evaluations

Gait Normal 10 10 10 10

Ataxia 0 0 0 0

Body Drags 0 0 0 0

Limbs splayed or dragging 0 0 0 0

Hunched posture 0 0 0 0

Locomotion Normal 10 10 10 9

Increased movement 0 0 0 1

Decreased movement 0 0 0 0

None 0 0 0 0

Arousal Normal 10 10 10 9

Low, Slight Stupor 0 0 0 0

Very Low, Stupor 0 0 0 0

High, Slight Excitement, Tense 0 0 0 1

Very High, Hyper

- Alert 0 0 0 0

Exophthalmia Not Present 10 10 10 10

Present 0 0 0 0

Piloerection Not Present 10 10 10 10

Present 0 0 0 0

Feces Number of pellets 0 2 0 0

Unformed Stool 0 0 0 0

Urine Number of Pools 3 2 0 0

Polyuria 0 0 0 0

Fasciculation Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Tremors Not Pr esent 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Convulsions Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Reflex Assessments

Visual Approach Normal, slowly approaches 10 10 10 10

No Reaction 0 0 0 0

Freezes or pulls away slightly 0 0 0 0

Jumps or Turns Abruptly to Avoid,

or attacks and/or bites 0 0 0 0

Audition Assessment N ormal 10 10 10 10

No Reaction 0 0 0 0

Exaggerated, Jumps, Flips, Bite 0 0 0 0

Pain Perception Normal 10 10 10 10

No Reaction 0 0 0 0

Increased Pain 0 0 0 0

Pupil Response Pupil Constricts Normally 10 10 10 10

Pupil Size does not Change 0 0 0 0

Miosis 0 0 0 0

Mydriasis 0 0 0 0

Proprioception Returns leg to original position 10 10 10 10

Returns leg only partially to original

position 0 0 0 0

No response 0 0 0 0

Pinna Reflex Ear flattens against head 0 1 0 0

Animals shakes head 10 9 10 10

No response 0 0 0 0

Air Righting Reflex Lands on All Four Feet (Normal) 10 10 10 10

Slightly Uncoordinated 0 0 0 0

Lands on Side 0 0 0 0

Lands on Back 0 0 0 0

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*Significantly different from control mean; p ≤ 0.05.

**Significantly dif ferent from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean. Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Body Weight (grams) Mean 555.6 557.0 578.1 545.7

S.D. 59.6 49.3 44.9 53.7

N 10 9 10 10

Forelimb Grip Strength (grams of force)

Mean 1284.8 1196.4 1178.8 1309.3

S.D. 293.1 2 24.5 187.4 228.1

N 10 9 10 10

Hindlimb Grip Strength (grams of force)

Mean 1003.0 946.9 992.5 911.3

S.D. 225.8 226.3 328.0 325.9

N 10 9 10 10

Landing Foot Splay (centimeters)

Mean 6.3 6.2 5.7 5. 7

S.D. 1.6 1.1 1.1 1.1

N 10 9 10 10

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 9 10 10

Home Cage Evaluations

Posture Sitting or Standing Normal ly 8 6 9 6

Alert, Oriented Toward Observer 2 3 1 4

Asleep 0 0 0 0

Lying on Side 0 0 0 0

Flattened 0 0 0 0

Crouched 0 0 0 0

Vocalizations Not Present 10 9 10 10

Present 0 0 0 0

Palpebral Closure Eyelids Open 10 9 10 10

Eyelids Completely Closed 0 0 0 0

Eyelids Slightly Drooping 0 0 0 0

Eyelids Half Closed 0 0 0 0

Handling Evaluations

Ease of Removal Very Easy 9 8 9 6

Easy 0 1 1 3

Moderately Difficult 1 0 0 1

Freezes 0 0 0 0

Difficult 0 0 0 0

Eas e of Handling Normal 0 1 0 0

Moderately Difficult 9 8 10 9

Freezes 1 0 0 1

Difficult 0 0 0 0

Totally limp 0 0 0 0

Chromodacryorrhea Not Present 10 9 10 10

Present 0 0 0 0

Lacrimation No lacrimation 10 9 10 10

Some lacrimation 0 0 0 0

Extreme lacrimation 0 0 0 0

Salivation Not Present 10 9 10 10

Slight 0 0 0 0

Moderate 0 0 0 0

Extreme 0 0 0 0

Coat Normal 10 9 10 10

Slightly Soiled 0 0 0 0

Moderately Soiled 0 0 0 0

Extremely Soiled 0 0 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Open Field Evaluations

Gait Normal 10 9 10 10

Ataxia 0 0 0 0

Body Drags 0 0 0 0

Limbs splayed or dragging 0 0 0 0

H unched posture 0 0 0 0

Locomotion Normal 10 9 10 10

Increased movement 0 0 0 0

Decreased movement 0 0 0 0

None 0 0 0 0

Arousal Normal 10 9 10 10

Low, Slight Stupor 0 0 0 0

Very Low, Stupor 0 0 0 0

High, Slight Excitement, Tense 0 0 0 0

Very High, Hyper

- Alert 0 0 0 0

Exophthalmia Not Present 10 9 10 10

Present 0 0 0 0

Piloerection Not Present 10 9 10 10

Present 0 0 0 0

Feces Number of pellets 10 24 13 6

Unformed Stool 0 0 0 0

Urine Number of Pools 13 11 7 5

Polyuria 0 0 0 0

Fasciculation Not Present 10 9 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Tremors Not Present 10 9 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Convulsions Not Present 10 9 10 10

Slight 0 0 0 0

Extr eme 0 0 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 9 10 10

Reflex Assessments

Visual Approach Normal, slowly approaches 9 9 9 10

No Reaction 0 0 1 0

Freez es or pulls away slightly 1 0 0 0

Jumps or Turns Abruptly to Avoid, or

attacks and/or bites 0 0 0 0

Audition Assessment Normal 10 9 10 10

No Reaction 0 0 0 0

Exaggerated, Jumps, Flips, Bite 0 0 0 0

Pain Perception Normal 10 9 10 10

No Reaction 0 0 0 0

Increased Pain 0 0 0 0

Pupil Response Pupil Constricts Normally 10 9 10 10

Pupil Size does not Change 0 0 0 0

Miosis (bilateral) 0 0 0 0

Mydriasis 0 0 0 0

Proprioception Returns leg to original position 10 9 10 1 0

Returns leg only partially to original

position 0 0 0 0

No response 0 0 0 0

Pinna Reflex Ear flattens against head 0 0 0 0

Animals shakes head 10 9 10 10

No response 0 0 0 0

Air Righting Reflex Lands on All Four Feet (Normal) 10 9 10 10

Slightly Uncoordinated 0 0 0 0

Lands on Side 0 0 0 0

Lands on Back 0 0 0 0

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*Significantly different from control mean; p ≤ 0.05.

**Significantly different from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean. Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Body Weight (grams) Mean 328.8 314.1 304.7 304.9

S.D. 36.3 29.2 35 .5 23.7

N 10 10 10 10

Forelimb Grip Strength (grams of force)

Mean 1092.8 1022.8 1150.3 1038.0

S.D. 206.2 197.0 126.1 177.4

N 10 10 10 10

Hindlimb Grip Strength (grams of force)

Mean 877.8 782.3 849.5 804.3

S.D. 190.4 176.3 142.7 90.5

N 10 10 10 10

Landing Foot Splay (centimeters)

Mean 4.2 4.6 4.7 4.9

S.D. 0.7 0.9 0.5 0.5

N 10 10 10 10

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13 th 

# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/ m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Home Cage Evaluations

Posture Sitting or Standing Normally 8 3 9 6

Alert, Oriented Toward Observer 2 7 1 4

Asleep 0 0 0 0

Lying on Side 0 0 0 0

Flattened 0 0 0 0

Crouched 0 0 0 0

Vocalizations Not Present 10 10 10 10

Present 0 0 0 0

Palpebral Closure Eyelids Open 10 10 10 10

Eyelids Completely Closed 0 0 0 0

Eyelids Slightly Drooping 0 0 0 0

Eyelids Half Closed 0 0 0 0

Handling Eval uations

Ease of Removal Very Easy 10 9 10 10

Easy 0 1 0 0

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Ease of Handling Very easy; totally limp 1 1 1 1

Normal 9 9 9 9

Moderately Difficult 0 0 0 0

Freezes 0 0 0 0

Difficult 0 0 0 0

Chromodacryorrhea Not Present 10 10 10 10

Present 0 0 0 0

Lacrimation No lacrimation 10 10 10 10

Some lacrimation 0 0 0 0

Extreme lacrimation 0 0 0 0

Salivation Not Present 10 10 10 10

Slight 0 0 0 0

Moderate 0 0 0 0

Extreme 0 0 0 0

Coat Normal 10 10 10 10

Slightly Soiled 0 0 0 0

Moderately Soiled 0 0 0 0

Extremely Soiled 0 0 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Anima ls Examined 10 10 10 10

Open Field Evaluations

Gait Normal 10 10 10 10

Ataxia 0 0 0 0

Body Drags 0 0 0 0

Limbs splayed or dragging 0 0 0 0

Hunched posture 0 0 0 0

Locomotion Normal 10 9 10 9

Increased movement 0 1 0 1

Decre ased movement 0 0 0 0

None 0 0 0 0

Arousal Normal 10 9 10 9

Low, Slight Stupor 0 0 0 0

Very Low, Stupor 0 0 0 0

High, Slight Excitement, Tense 0 1 0 1

Very High, Hyper

- Alert 0 0 0 0

Exophthalmia Not Present 10 10 10 10

Present 0 0 0 0

Piloerection Not Present 10 10 10 10

Present 0 0 0 0

Feces Number of pellets 6 0 4 0

Unformed Stool 0 0 0 0

Urine Number of Pools 6 3 5 4

Polyuria 0 0 0 0

Fasciculation Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Tremors Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

Convulsions Not Present 10 10 10 10

Slight 0 0 0 0

Extreme 0 0 0 0

# Huntingdon Life Sciences 0 0

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# week of exposure Table 9

Group I Group II Group III Group IV

Exposure Level (mg/m 3 

# ): 0 2000 10000 20000

Number of Animals Examined 10 10 10 10

Reflex Assessments

Visual Approach Normal, slowly approaches 10 10 10 10

No Reaction 0 0 0 0

Freezes or pulls away slightly 0 0 0 0

Jumps or Turns Abruptly to Avoid,

or attacks and/or bites 0 0 0 0

Audition Assessment Normal 10 10 10 10

No Reaction 0 0 0 0

Exaggerated, Jumps, Flips, Bite 0 0 0 0

Pain Perception Normal 10 10 10 10

No Reaction 0 0 0 0

Increased Pain 0 0 0 0

Pupil Response Pupil Constricts Normal ly 10 10 10 10

Pupil Size does not Change 0 0 0 0

Miosis (bilateral) 0 0 0 0

Mydriasis 0 0 0 0

Proprioception Returns leg to original position 10 10 10 10

Returns leg only partially to

original position 0 0 0 0

No response 0 0 0 0

Pinna Reflex Ear flattens against head 0 0 0 1

Animals shakes head 10 10 10 9

No response 0 0 0 0

Air Righting Reflex Lands on All Four Feet (Normal) 10 10 10 10

Slightly Uncoordinated 0 0 0 0

Lands on Side 0 0 0 0

Lands on Back 0 0 0 0

# Huntingdon Life Sciences 00

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Mean Hematology Values

Preface Table 10

Reporting

Abbreviation Parameter Units

HGB Hemoglobin Concentration g/dL

HCT Hematocrit percent

RBC Erythrocyte Count 10 6 

# / μ L

RETIC Absolute Reticulocyte Count 10 9 

# /L

PLT Platelet Count 10 3 

# / μ L

MCV Mean Corpuscular Volume fL

MCH Mean Corpuscular Hemoglobin pg

MCHC Mean Corpuscular Hemo globin

Concentration g/dL

WBC Total Leukocyte Count 10 3 

# / μ L

ANEU Absolute Neutrophils 10 3 

# / μ L

ALYM Absolute Lymphocytes 10 3 

# / μ L

AMONO Absolute Monocytes 10 3 

# / μ L

AEOS Absolute Eosinophils 10 3 

# / μ L

ABASO Absolute Basophils 10 3 

# / μ L

ALUC Absolute Large Unstained Cells 10 3 

# / μ L

Key to Statistical Symbols:

*Significantly different from control mean; p ≤ 0.05.

**Significantly different from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control meanHuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-Week4GroupHGBHCTg/dL%1M0mg/m(3)Mean16.048.4SD0.541.43n10 102M2000mg/m(3)Mean15.848.4SD0.361.47n10 103M10000mg/m(3)Mean15.547.2SD0.422.02n10 104M**20000mg/m(3)Mean15.446.9SD0.511.70n10 10RBCX10A6/UL8.620.294108.730.392108.460.481108.550.30910RETICPLTMCVMCHMCHCWBCX10A9/LX10A3/ULfLpgg/dLX10A3/uL202.490256.218.533.016.0030.41146.41.390.470.502.13610 1010101010*177.191455.618.232.716.5019.35 194.91.650.610.332.64110 10 1010 10 10**168.891555.818.432.917.1625.48215.01.480.78 0.793.406101010101010**155.893054.918.032.714.6226.41287.21.150.460.462.39210 10 10101010Page 180HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-Week4GroupANEUALYM1Momg/m(3)Mean1.9413.21SD0.4372.022n10102M2000mg/m(3)Mean1.7813.96SD0.5552.674n10103M10000mg/m(3)Mean1.6714.69SD0.6052.744n10104M20000mg/m(3)Mean2.0211.81SD0.8242.062n1010AMONOAEOSABASOALUC0.42 0.22 0.140.070.1050.1020.0600.041101010100.36 0.15 0.160.090.1520.0500.0450.026101010100.390.17 0.15 0.110.168 0.1390.0450.035101010100.330.230.130.100.1600.1740.0350.04010101010Page 181HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-Week4GroupHGBHCTg/dL%1F0mg/m(3)Mean15.646.6SD0.471.42n10102F2000mg/m(3)Mean15.947.9SD0.853.54n10 103F10000mg/m(3)Mean15.245.7SD0.572.19n10 104F20000mg/m(3)Mean15.145.2SD0.712.51n10 10RBCX10A6/uL8.380.282108.710.687108.180.464108.270.55510RETICPLTMCVMCHMCHCWBCX10A9/Lx10A3/ULfLpgg/dLX10A3/UL194.4100055.618.633.515.7233.69153.01.140.350.333.5441010 10101010189.696955.018.333.214.9430.79190.61.780.87 0.962.40110 1010101010188.995155.918.633.415.4135.92297.81.040.550.613.381101010101010185.1104854.718.333.414.1536.99229.92.150.650.562.460101010101010Page 182HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-Week4GroupANEUALYMX10A3/uLX10A3/uL1F0mg/m(3)Mean1.7813.15SD0.4073.106n10102F2000mg/m(3)Mean1.4012.80SD0.4202.068n10103F10000mg/m(3)Mean1.7912.87SD0.5352.790n10104F20000mg/m(3)Mean1.6311.88SD0.6892.096n1010AMONOAEOSABASOALUCX10A3/uLx10A3/uLx10A3/uLx10A3/UL0.390.150.150.100.0750.0770.0580.044101010100.310.210.130.100.1120.1970.0360.025101010100.310.210.120.100.1180.1880.0450.03310101010**0.250.190.110.090.100·0.1360.0270.03410101010Page 183HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-TerminationGroupHGBHCTg/dL%1M0mg/m(3)Mean16.248.1SD0.581.63n10102M2000mg/m(3)Mean16.247.8SD0.551.67n993M10000mg/m(3)Mean15.746.8SDl.082.90n10104M20000mg/m(3)Mean16.047.7SD0.792.76n1010RBCRBTICX10A6/ULX10A9/L9.16189.20.36730.6910109.15169.90.39519.11998.97202.00.65594.131010**9.26150.10.47718.811010PLTMCVMCHMCHCWBCX10A3/ULfLpgg/dLX10A3/UL85552.617.733.712.08282.91.520.540.602.8741010101010*112552.217.733.913.28143.21.210.530.684.34799999*112352.217.533.611.10197.81.681.191.892.0421010101010*104451.517.333.611.31241.81.010.410.702.9921010101010Page 184HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-TerminationGroupANEUALYM1M0mg/m(3)Mean2.269.14SD0.7092.501n10102M2000mg/m(3)Mean2.1010.54SD0.9253.844n993M10000mg/m(3)Mean1.778.81SD0.5661.713n10104M20000mg/m(3)Mean1.808.95SD0.5372.672n1010AMONOAEOSABASOALUC0.330.190.110.050.1000.1070.0910.032101010100.310.160.110.080.1610.0690.0590.05499990.270.140.070.050.1790.0910.0200.013101010100.280.20o.os0.060.1670.1220.0520.03910101010Page 185HuntingdonLifesciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-TerminationGroupHGBHCTg/dL%1F0mg/m(3)Mean15.645.5SD0.461.42n10102F2000mg/m(3)Mean16.147.2SD0.913.34n10103F10000mg/m(3)Mean15.946.0SD0.521.69n10104F20000mg/m(3)Mean15.846.5SD0.551.81n99RBC8.330.278108.690.662108.390.415108.610.4149RETICPLTMCVMCHMCHCWBCfLpgg/dL171.3102454.618.734.29.9029.67224.51.250.540.333.189101010101010171.792154.418.634.29.5622.32158.21.730.770.882.268101010101010179.6101354.819.034.69.8834.10232.11.280.400.562.805101010101010193.3102054.018.434.09.2334.14243.62.540.840.411.807999999Page 186HuntingdonLifesciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-TerminationGroupANEUALYM1Forng/rn(3)Mean1.887.54SD1.0702.575n10102F2000rng/rn(3)Mean1.068.10SD0.3032.021n10103F10000rng/rn(3)Mean1.557.78SD0.6372.110n10104F20000rng/rn(3)Mean1.347.46SD0.5591.406n99AMONOAEOSABASOALUC0.240.150.050.040.1170.1010.0270.025101010100.180.130.050.050.0790.0490.0250.032101010100.210.220.070.050.1170.2010.0330.013101010100.160.140.050.040.0530.0590.0320.0289999Page 187HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-RecoveryGroup1M0mg/m(3)MeanSDn4M20000mg/m(3)MeanSDnHGBg/dL15.50.551015.50.5210HCT%48.12.231047.21.8410RBC9.100.446108.990.41610RETIC162.434.8110**126.812.3410PLT983196.2101020238.510MCVfL52.90.881052.61.6810MCHpg17.10.941017.31.2210MCHCg/dL32.41.501032.91.5610WBC12.623.3701012.023.13110Page 188HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-RecoveryGroup1M0mg/m(3)MeanSDn4M20000mg/m(3)MeanSDnANEUX10A3/uL1.780.508101.960.83410ALYMX10A3/UL9.922.938109.323.28410AMONOx10A3/uL0.460.175100.350.11610AEOSX10A3/uL0.210.185100.170.13610ABASOX10A3/uL0.200.103100.160.06010ALUCX10A3/uL0.050.034100.060.02510Page 189HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-RecoveryGroup1F0mg/m(3)MeanSDn4F20000mg/m(3)MeanSDnHGBg/dL15.30.661015.60.4410HCT%45.82.271046.91.9610RBCX10A6/UL8.260.314108.500.35510RETICx10A9/L133.026.0210118.524.2510PLTX10A3/UL1044180.110878225.710MCVfL55.41.221055.11.2710MCHpg18.50.511018.30.5110MCHCg/dL33.50.571033.20.7510WBCx10A3/uL10.621.4091010.482.09810Page 190HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable10MeanHematologyValues-RecoveryGroup1Fomg/m(3)MeanSDn4F20000mg/m(3)MeanSDnANEU1.710.574101.520.68510ALYM8.301.345108.361.42510AMONO0.340.11010**0.220.04910AEOS0.110.03310*0.190.15510ABASO0.120.040100.160.06810ALUC0.040.012100.040.01410Page 191

# Huntingdon Life Sciences 00

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Mean Coagulation Values

Preface Table 11

Reporting

A bbreviation Parameter Units

PT Prothrombin Time Seconds

APTT Activated Partial Thromboplastin Time Seconds

Key to Statistical Symbols:

*Significantly different from control mean; p ≤ 0.05.

**Significantly different from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean.HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable11MeanCoagulationValues-Week4GroupPTAPTTSeconds Seconds1Momg/m(3)Mean11.9 17.7SD0.502.67n10102M***2000mg/m(3)Mean12.520.4SD0.341.50n10103M***10000mg/m(3)Mean12.620.4SD0.641.69n10104M***20000mg/m(3)Mean12.521.1SD0.742.04n1010Page 193HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable11MeanCoagulationValues-Week4GroupPTAPTTSecondsSeconds1Fomg/m(3)Mean11.419.8SD0.461.26n10102F2000mg/m(3)Mean11.519.2SD0.581.71n993F10000mg/m(3)Mean11.619.5SD0.332.63n10104F20000mg/m(3)Mean11.619.8SD0.461.16n1010Page 194HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable11MeanCoagulationValues-TerminationGroupPTAPTTSeconds Seconds1M0mg/m(3)Mean12.2 19.2SD0.443.16n10102M2000mg/m(3)Mean12.721.2SD0.582.27n993M10000mg/m(3)Mean12.7 19.8SD1.063.03n10104M20000mg/m(3)Mean12.520.5SD00722.67n1010Page 195HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable11MeanCoagulationValues-TerminationGroupPTAPTTSecondsSeconds1F0mg/m(3)Mean11.218.2SD0.272.99n10102F2000mg/m(3)Mean11.116.6SD0.371.45n10103F10000mg/m(3)Mean11.217.2SD0.292.21n10104F20000mg/m(3)Mean11.216.9SD0.381.78n1010Page 196

# Huntingdon Life Sciences 00

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Mean Clinical Chemistry Values

Preface Table 12

Reporting

Abbreviation Parameter Units

AST Aspartate Aminotransferase U/L

ALT Alanine Aminotransferase U/L

ALKP Alkaline Phosphatase U/L

LD Lactate Dehydrogenase U/L

SDH Sorbitol Dehydrogenase U/L

BU N Blood Urea Nitrogen mg/dL

CREAT Creatinine mg/dL

GLU Fasting Glucose mg/dL

CK Creatine Kinase U/L

CHOL Cholesterol (Enzymatic) mg/dL

TP Total Protein g/dL

ALB Albumin g/dL

GLOB Globulin (calculated) g/dL

A/G Albumin/Globulin Ratio (calculated)

TBILI Tota l Bilirubin mg/dL

DBILI Direct Bilirubin mg/dL

IBILI Indirect Bilirubin (calculated) mg/dL

Na + 

# Sodium mEq/L

K + 

# Potassium mEq/L

Cl

- 

# Chloride mEq/L

Ca ++ 

# Calcium mg/dL

PHOS Inorganic Phosphorus mg/dL

GGT Gamma

- Glutamyl Transferase U/L

Bile Acids Bile Acids μ m ol/L

Key to Statistical Symbols:

*Significantly different from control mean; p ≤ 0.05.

**Significantly different from control mean; p ≤ 0.01.

If no asterisks, no statistically significant differences from control mean.HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-Week4GroupASTALTU/LU/L1M0mg/m(3)Mean8144SD9.76.9n10102M2000mg/m(3)Mean7539SD2.86.2n10103M10000mg/m(3)Mean7840SD11.77.7n10104M**20000mg/m(3)Mean7l38SD8.85.3n1010ALKPLDU/LU/L15818535.8 31.41010*15813221.324.51010**156 11827.265.11010144**8922.721.11010SOHBUNCREATGLUCKU/Lmg/dLmg/dLmg/dLU/L18.0130.31272402.902.10.059.1191.81010101010**15.3120.3118 1512.771.80.038.728.31010101010 14.7120.31111493.761.80.067.662.o1010101010** ***14.4120.31141282.781.40.037.530.81010101010Page 198HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-Week4GroupCHOLTPmg/dLg/dL1M0mg/m(3)Mean666.5SD9.10.27n10102M2000mg/m(3)Mean616.4SD11.20.21n10103M10000mg/m(3)Mean576.4SD10.30.33n10104M20000mg/m(3)Mean596.4SD9.60.30n1010ALBg/dL4.10.20104.10.07104.10.16104.10.2410GlobA/GTBILIDBILIIBILI..Na+g/dLmg/dLmg/dL mg/dLmEq/L2.41.80.130.040.091480.170.170.0250.0080.0231.01010101010102.31.80.120.030.091480.180.130.0220.0090.0161.31010101010102.31.80.130.040.091470.210.150.0250.0070.0221.1101010101092.31.80.110.030.081480.170.150.0170.0090.0181.2101010101010Page 199HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-week4GroupK+Cl-Ca++PHOSGGTBileAcidsmEq/LmEq/Lmg/dLmg/dLU/Lumol/L1M0mg/m(3)Mean5.61031.1.510.0052SD0.400.70.241.000.641.8n10101.01.010102M2000mg/m(3)Mean5.51.0211.39.8077SD0.33l.l.0.230.410.085.1.n1.01.0101010103M10000mg/m(3)Mean5.41.0311.51.0.2065SD0.381.30.220.730.072.9n99991094M20000mg/m(3)Mean5.510311.39.8037SD0.502.00.330.670.025.4n10101010 101.0Page 200HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-Week4GroupASTALTU/LU/L1F0mg/m(3}Mean7736SD14.37.9n10102F2000mg/m(3}Mean7834SD13.a7.6n10103F10000mg/m(3}Mean7033SD6.34.0n10104F20000mg/m(3)Mean69*29SD3.9 3.7n1010ALKPLDU/LU/L958714.338.6101010811721.788.01010996830.716.91010896215.023.21010SDHBUNCREATGLUCKU/Lmg/dLmg/dLmg/dLU/L15.3150.51221282.261.10.074.854.3101010101015.614.0.4135 1655.972.30.0828.564.61010101010*11.5130.41171082.171.80.066.222.01010101010**12.0140.41161252.731.80.038.658.21010101010Page 201HuntingdonLifesciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-Week4GroupCHOLTPrng/dLg/dLlF0rng/rn(3)Mean817.4SD12.70.59n10102F2000rng/rn(3)Mean857.2SD11.80.36nB103F10000rng/rn(3)Mean957.4SD12.50.36n8104F20000rng/rn(3)Mean927.3SD18.80.49n810ALBg/dL4.80.42104.70.27104.80.23104.80.2910GlobA/GTBILIDBILIIBILINa+g/dLrng/dLrng/dLrng/dLrnEq/L2.61.90.180.040.131480.210.130.031000110.0281.01010101010102.51.90.170.040.131490.210.200.0400.0140.0273.3101010101010**2.61.90.170.040.131520.190.120.0330.0160.0231.1101010101010**2.51.90.150.030.111520.250.150.0370.0110.0341.6101010101010Page 202HuntingdonLifeSciencesStudyNo.00-6130sponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-Week4GroupK+Cl-Ca++PHOSGGTBileAcidsmEq/LmEq/Lmg/dL mg/dLU/Lumol/L1Fomg/m{3)Mean5.310211.99.9084SD0.521.40.450.560.059.0n101010101092F***2000mg/m{3)Mean6.010412.010.8026SD0.543.40.241.730.318.9n101010101093F 10000mg/m{3)Mean5.810512.010.2045SD0.311.80.200.850.051.1n101010101084F** **20000mg/m{3)Mean5.610611.89.5034SD0.371.80.330.71o.o19.8n10101010109Page 203HuntingdonLifesciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryvalues-TerminationGroupASTALTU/LU/L1Morng/m(3)Mean8544SD11.46.4n10102M2000rng/rn(3)Mean8545SD14.56.7n993M10000rng/rn(3)Mean8441SD14.57.4n10104M20000rng/rn(3)Mean7238SD13.18.4n1010ALKPU/L9015.2108914.899313.9108516.910LDSOHBUNCREATGLUCKU/LU/Lrng/dLrng/dLmg/dLU/L18117.6150.4149190117.64.482.00.0724.9124.410101010101024617.0150.5165234159.05.191.30.0752.5185.299999918915.9140.4131193128.55.131.40.1220.4121.810101010101011315.4150.413613543.56.081.70.0734.246.4101010101010Page 204HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryvalues-TerminationGroupCHOLTPmg/dLg/dL1M0mg/m(3)Mean697.1SD14.30.35n10102M2000mg/m(3)Mean647.0SD17.00.30n993M10000mg/m(3)Mean667.0SD15.60.30n10104M20000mg/m(3)Mean616.8SD17.40.40n1010ALBGlobg/dLg/dL4.32.80.230.1910104.32.70.190.23994.42.60.080.2510104.22.60.200.241010A/GTBILIDBILIIBILINa+mg/dL mg/dL mg/dLmEq/L1.50.110.030.081480.100.0170.0190.0191.110101010101.60.110.020.091480.160.0250.0090.0190.799999*1.70.100.030.071480.160.0150.0130.0151.71010101010*1.60.090.020.071480.130.0210.0090.0181.41010101010Page 205HuntingdonLifeSciencesStudyNo.SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-TerminationGroupK+Cl-Ca++PHOSGGTBileAcidsmEq/L mEq/Lmg/dLmg/dLU/Lumol/L1Momg/m(3)Mean5.911.38.4027SD0.372.80.650.960.017.2n1010101010102M2000mg/m(3)Mean5.710111.18.9023SD0.532.20.612.290.016.5n9999993M10000mg/m(3)Mean5.810211.18.6028SD0.581.80.322.040.017.2n1010101010104M20000mg/m(3)Mean5.910210.98.8021SD0.583.00.211.520.67.9n101010101010Page 206HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-TerminationGroupASTALTU/LU/L1F0mg/m(3)Mean9142SD18.610.7n10102F2000mg/m(3)Mean8436SD9.811.1n10103F**10000mg/m(3)Mean7133SD9.63.7n10104F**20000mg/m(3)Mean7536SD11.913.4n1010ALKPU/L439.7104811.2104312.210445.910LDSDHBUNCREATGLUCKU/LU/Lmg/dLmg/dL mg/dLU/L27018.2150.5140210114.34.671.80.0625.996.510101010101031815.1170.5135184163.62.321.90.0727.271.9101010101010 12315.1140.513612440.33.571.50.0619.065.7101010101010**  ****8612.7160.51298731.34.051.90.0317.322.0101010101010Page 207HuntingdonLifesciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-TerminationGroupCHOLTPmg/dLg/dL1Fomg/m(3)Mean777.7SD14.10.71n10102F2000mg/m(3)Mean807.7SD20.20.47n10103F10000mg/m(3)Mean907.7SD17.60.64n10104F20000mg/m(3)Mean917.9SD13.00.45n1010ALBg/dL5.10.49105.10.44105.10.46105.10.3910GlobA/GTBILIDBILIIBILINa+g/dLmg/dLmg/dLmg/dLmEq/L2.71.90.140.030.111480.310.190.0360.0140.0282.21010101010102.52.00.150.030.121480.180.240.0390.0130.0311.8101010101010*2.71.90.130.020.111500.310.270.0130.0110.0091.5101010101010**2.81.80.130.030.101500.170.150.0220.0080.0212.1101010101010Page 208HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-TerminationGroupK+Cl-Ca++PHOSGGTBileAcidsmEq/LmEq/Lmg/dLmg/dLU/Lumol/L1F0mg/m(3)Mean4.810311.36.8037SD0.633.30.391.41o.o27.9n1010101010102F**2000mg/m(3)Mean5.710311.66.8033SD0.493.40.510.90o.o26.9n1010101010103F**10000mg/m(3)Mean5.510311.67.2025SD0.472.40.261.320.014.5n1010101010104F20000mg/m(3)****Mean6.010311.97.1054SD0.483.70.460.910.454.0n101010101010Page 209HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-RecoveryGroup1M0mg/m(3)4M20000mg/m(3)MeanSDnMeanSDnASTU/L8940.710*729.210ALTU/L5122.710439.110ALKPU/L8313.710866.310LDU/L209175.310**9434.610SDHU/L19.65.3810*14.72.6410BUNmg/dL131.010131.510CREATmg/dL0.40.05100.3o.os10GLUmg/dL11830.51011327.610CKU/L15485.61010046.710Page 210HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPB-STable12MeanClinicalChemistryValues-RecoveryGroup1Momg/m(3)MeanSDn4M20000mg/m(3)MeanSDnCHOLmg/dL6713.2106719.010TPg/dL7.00.23106.90.1610ALBg/dL4.20.17104.20.1410Globg/dL2.80.20102.70.1810A/G1.50.14101.60.1510TBILImg/dL0.130.019100.130.01310DBILImg/dL0.040.011100.030.01210IBILImg/dL0.090.017100.100.01510Na+mBq/L1461.810**1481.010Page 211HUntingdonLifeSciencesstudyNo.oo-6130SponsorStudyNo.211-DIPB-STable12MeanClinicalChemistryvalues-RecoveryGroup1M0mg/m(3)4M20000mg/m(3)MeanSDDMeanSDnK+mBq/L6.10.51106.20.2410ClmBq/L1041.510**1051.310C&++mg/dL11.20.281011.10.4310PHOSmg/dL7.80.64107.40.3910GGTU/L00.41000.310BileAcidsumol/L3924.2103111.210Page 212HuntingdonLifeSciencesStudyNo.00-6130SponsorStudyNo.211-DIPB-STable12MeanClinicalChemistryvalues-RecoveryGroup1F0mg/m(3l4F20000mg/m(3lMeanSDnMeanSDnASTU/L10975.7108129.610ALT0/L6242.0105126.310ALKPU/L4111.8104010.410LDU/L14954.910**6919.910SOHU/L19.48.2110*12.85.2510BUNmg/dL141.810152.310CRBATmg/dL0.40.05100.5o.os10GLUmg/dL997.41010310.910CK0/L10320.410*7722.410Page 213HuntingdonLifeSciencesStudyMo.00-6130SponsorStudyMo.211-DIPE-STable12MeanClinicalChemistryValues-RecoveryGroup1F0mg/m(3)4F20000mg/ml3)MeanSDnMeanSDnCHOLmg/dL9620.1109619.210TPg/dL7.60.62107.90.4410ALBg/dL5.00.47105.10.4510Globg/dL2.60.25102.90.3410A/G1.90.21101.90.2810TBILImg/dL0.190.025100.170.04010DBILImg/dL0.040.014100.040.01810IBILImg/dL0.130.019100.130.02510Ma+mEq/L1491.610*1491.410Page 214HUntingdonSciencesStudyNo.00-6130SponsorStudyNo.211-DIPE-STable12MeanClinicalChemistryValues-RecoveryGroup1F0mg/m(3)4F20000mg/m(3)MeanSDnMeanSDn!!:+5.70.34105.50.3610Cl1062.9101071.610Ca++11.60.421011.80.4410PHOS6.50.60106.20.7110GGTU/L00.71000.710BileAcidsumol/I.3421.6105457.510Page 215

# Huntingdon Life Sciences 00

- 6130 Page 216

211

- DIP E

- S Final Report

Mean Organ Weights and

Brain Measurements

Preface Table 13

Table of Contents

Absolute Organ Weights (Terminal Sacrifice

- Main Study)

.............................. 217

% Organ to Body Weight (Terminal Sacrifice

- Main Study)

............................. 221

% Organ to Brain Weight (Terminal Sacrifice

- Main Study)

............................. 225

Neurotox Brain Measurements (Terminal Sacrifice)

......................................... 229

Absolute Organ Weights (Recovery Sacrifice

- Main Study)

............................ 231

% Organ to Body Weight (Recovery Sacrifice

- Main Study)

............................ 235

% Organ to Brain Weight (Recovery Sacrifice

- Main Study)

............................ 239

Note: Brain length and width were measured in centimeters.

Corresponding exposure levels for each group were as follows:

Group 1

- 0 mg/m 3



# Group 2

- 2000 mg/m 3



# Group 3

- 10000 mg/m 3



# Group 4

- 20000 mg/m 3



# Key to Abbreviations:

g = Grams

wt. = Weight

observ. = ObservedHuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:MainStudy-TerminalSacrificeGroupSummarystatisticsforabsoluteorganweights(g)Studynumber:006130BrainHeartTerminalBodywt.(g)AdrenalGlandsEpididymisKidneys1Me an:Standarddeviation:Numberofobserv.:2Me an:Standarddeviation:Numberofobserv.:3Me an:Standarddeviation:Numberofobserv.:4Me an:Standarddeviation:Numberofobserv.:528.921.9(10)506.235.2(9)514.446.0(10)516.658.7(10)0.07250.0066(10)0.07260.0105(9)0.06800.0123(10)0.07010.0080(10)Ma1e2.21550.0985(10)2.14770.1038(9)2.16740.0766(10)2.16390.0921(10)Anim a1s1.57650.1600(10)1.63470.2308(9)1.43790.4102(10)1.62590.1776(10)1.74220.1418(10)1.65660.1391(9)1.66140.1786(10)1.64370.2459(10)4.34640.4963(10)4.25920.3751(9)4.47380.4413(10)4.56840.4495(10)Printed:30-Mar-04Page:1Liver15.83451.6854(10)14.70901.9306(9)15.04971.5655(10)15.53932.5562(10)Version4.2.2Lungs2.30080.3150(10)2.09400.1722(9)2.15000.2456(10)2.0055*0.2828(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%{$)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 217HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseySummarystatisticsforabsoluteorganweights(g)Studynumber:006130Table13:MainStudy-TerminalSacrificeGroup1M e an:Standarddeviation:Numberofobserv.:2M e an:Standarddeviation:Numberofobserv.:3M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)528.921.9(10)506.235.2(9)514.446.0(10)516.658.7(10)SeminalvesiclesProstateM a 1 e1.13311.72790.28700.2915(10)(10)1.07661.42150.18930.2231(9)(9)1.24921.75060.25330.5404(10)(10)1.15171.74140.23880.3770(10)(10)TestesSpleenA n i m a 1 s0.77443.60930.12090.5810(10)(10)0.84193.42850.12720.3058(9)(9)0.79513.41890.13150.8429(10)(10)0.77523.73690.15360.2558(10)(10)Thymus0.36970.0893(10)0.39020.0750(9)0.35630.0638(10)0.33460.0674(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 218HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:MainStudy-TerminalSacrificeGroupSummarystatisticsforabsoluteorganweights(g)Studynumber:006130BrainKidneysTerminalBodywt.(g)AdrenalGlandsHeart1Me an:Standarddeviation:Numberofobserv.:2M e an:Standarddeviation:Numberofobserv.:3M e an:Standarddeviation:Numberofobserv.:4Me an:Standarddeviation:Numberofobserv.:294.024.5(10)288.525.2(10)309.132.0(10)284.628.3(10)0.06830.0078(10)0.07680.0118(10)0.06750.0151(10)0.06960.0115(10)F e m a l e2.02820.0739(10)2.06830.1021(10)2.10270.0831(10)2.07940.0692(10)Anim als1.05650.1207(10)1.08220.1119(10)1.11830.1006(10)1.00790.0943(10)2.36020.1791(10)2.39570.3473(10)2.65430.2585(10)2.42400.2768(10)Liver8.31270.9646(10)8.38211.0932(10)9.16851.5541(10)8.82501.0968(10)Printed:30-Mar-04Page:1Lungs1.59540.1767(10)1.61580.2393(10)1.73110.1689(10)1.60560.2027(10)Version4.2.2Ovaries0.09290.0152(10)0.09210.0140(10)0.09150.0157(10)0.08900.0132(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 219HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:MainStudy-TerminalSacrificeGroup1M e an:Standarddeviation:Numberofobserv.:2M e an:Standarddeviation:Numberofobserv.:3M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)294.024.5(10)288.525.2(10)309.132.0(10)284.628.3(10)Summarystatisticsforabsoluteorganweights(g)Studynumber:006130ThymusSpleenUterusF e m a 1 eA n i m a 1 s0.57680.31900.89840.05570.04830.1815(10)(10)(10)0.55020.33370.6860$0.08890.05170.0609(10)(10)(10)0.58810.33921.00490.08740.06120.3861(10)(10)(10)0.57710.29910.80030.07550.06880.2566(10)(10)(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 220HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:MainStudy-TerminalSacrificeGroupSummaryStatisticsfor%OrgantoBodyWeightStudynumber:006130BrainHeartTerminalBodywt.(g)AdrenalGlandsEpididymis1Me an:Standarddeviation:Numberofobserv.:2Me an:Standarddeviation:Numberofobserv.:3Me an:Standarddeviation:Numberofobserv.:4Me an:Standarddeviation:Numberofobserv.:528.921.9(10)506.235.2(9)514.446.0(10)516.658.7(10)0.01370.0012(10)0.01450.0027(9)0.01320.0017(10)0.01360.0015(10)Ma1e0.41900.0135(10)0.42580.0310(9)0.42340.0284(10)0.42300.0429(10)Anim a1s0.29800.0268(10)0.32290.0390(9)0.28000.0837(10)0.31720.0419(10)0.32950.0245(10)0.32770.0235(9)0.32320.0235(10)0.31740.0167(10)Kidneys0.82110.0801(10)0.84130.0437(9)0.87140.0715(10)0.88660.0521(10)Printed:30-Mar-04Page:1Liver2.99170.2792(10)2.89810.2180(9)2.92680.1934(10)2.99570.2264(10)Version4.2.2Lungs0.43450.0510(10)0.41540.0435(9)0.41800.0313(10)0.3876*0.0225(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 221HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseySummaryStatisticsfor%OrgantoBodyWeightStudynumber:006130Table13:MainStudy-TerminalSacrificeGroup1M e an:Standarddeviation:Numberofobserv.:2M e an:Standarddeviation:Numberofobserv.:3M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)528.921.9(10)506.235.2(9)514.446.0(10)516.658.7(10)SeminalvesiclesProstateM a 1 e0.21490.32680.05610.0537(10)(10)0.21370.28050.04100.0365(9)(9)0.24470.34230.05460.1093(10)(10)0.22300.33870.03910.0722(10)(10)TestesSpleenA n i m a 1 s0.14660.68150.02290.0990(10)(10)0.16720.68000.02820.0737(9)(9)0.15520.66530.02590.1664(10)(10)0.14960.73200.02260.1040(10)(10)Thymus0.06950.0146(10)0.07670.0121(9)0.06940.0120(10)0.06460.0100(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 222HuntingdonLifeSciencesSummaryStatisticsfor%OrgantoBodyWeightPrincetonResearchCenterStudynumber:006130EastMillstone,NewJerseyTable13:MainStudy-TerminalSacrificeGroupTerminalBrainKidneysBodywt.(g)AdrenalGlandsHeartF e m a l eA n i m a l s1M e an:294.00.02340.69280.35960.8067Standarddeviation:24.50.00340.04630.03190.0829Numberofobserv.:(10)(10)(10)(10)(10)2M e an:288.50.02680.72060.37580.8300Standarddeviation:25.20.00450.05770.03140.0946Numberofobserv.:(10)(10)(10)(10)(10)3M e an:309.10.02190.68670.36290.8646Standarddeviation:32.00.00470.07610.02290.1009Numberofobserv.:(10)(10)(10)(10)(10)4M e an:284.60.02460.73630.35540.8542Standarddeviation:28.30.00440.06680.02830.0851Numberofobserv.:(10)(10)(10)(10)(10)Liver2.83320.3111(10)2.89980.1907(10)2.95820.2915(10)3.10460.2585(10)Printed:30-Mar-04Page:1Version4.2.2LungsOvaries0.54300.03180.04560.0060(10){10)0.56130.03220.08240.0058(10)(10)0.56390.02980.06620.0055(10)(10)0.56940.03150.09960.0055(10)(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%{$)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 223HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:MainStudy-TerminalSacrificeGroupTerminalBodywt.(g)1M e an:294.0Standarddeviation:24.5Numberofobserv.:(10)2M e an:288.5Standarddeviation:25.2Numberofobserv.:(10)3M e an:309.1Standarddeviation:32.0Numberofobserv.:(10)4M e an:284.6Standarddeviation:28.3Numberofobserv.:(10)SummaryStatisticsfor%OrgantoBodyWeightStudynumber:006130ThymusSpleenUterusF e m a l e A n i m a l s0.19740.10900.30670.02520.01810.0645(10)(10)(10)0.19030.11660.2383%0.02250.02140.0174(10)(10)(10)0.19040.10990.32850.02250.01740.1320(10)(10)(10)0.20380.10550.28520.02650.02700.0981(10)(10)(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 224HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:MainStudy-TerminalSacrificeGroupSummaryStatisticsfor%OrgantoBrainWeightStudynumber:006130BrainHeartTerminalBodywt.(g)AdrenalGlandsEpididymis1Me an:Standarddeviation:Numberofobserv.:2Me an:Standarddeviation:Numberofobserv.:3M e an:Standarddeviation:Numberofobserv.:4Me an:Standarddeviation:Numberofobserv.:528.921.9(10)506.235.2(9)514.446.0(10)516.658.7(10)3.27510.3068(10)3.39700.5725(9)3.14070.5634(10)3.24100.3416(10)Ma1e100.00000.0000(10)100.00000.0000(9)100.00000.0000(10)100.00000.0000(10)Anim a1s71.18616.7627(10)76.07549.9999(9)66.467919.1909(10)75.412810.1435(10)78.79237.5643(10)77.34408.0110(9)76.70958.2462(10)76.071011.9499(10)Kidneys196.387423.1171(10)198.433216.3400(9)206.509719.8247(10)211.086218.7674(10)Printed:30-Mar-04Page:1Liver714.107263.3307(10)686.091194.1824(9)694.302766.4631(10)718.6379120.2962(10)Version4.2.2Lungs103.879513.5444(10)97.57057.6301(9)99.11979.7955(10)92.693312.8593(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 225HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseySummaryStatisticsfor%OrgantoBrainWeightStudynumber:006130Table13:MainStudy-TerminalSacrificeGroup1M e an:Standarddeviation:Numberofobserv.:2M e an:Standarddeviation:Numberofobserv.:3M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)528.921.9(10)506.235.2(9)514.446.0(10)516.658.7(10)SeminalvesiclesProstateM a 1 e51.204378.002612.869612.6975(10)(10)50.162665.9880%8.78428.2619(9)(9)57.474381.118910.528625.8248(10)(10)53.455280.490211.9996 17.0347(10)(10)TestesSpleenA n i m a 1 s35.0394163.22565.860027.5023(10)(10)39.2541159.75286.088713.3620(9)(9)36.7023158.01055.954439.5019(10)(10)35.8259173.29076.951317.9686(10)(10)Thymus16.63053.6458(10)18.19423.6350(9)16.50363.2765(10)15.46633.0664(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 226HuntingdonLifeSciencesSummaryStatisticsfor%OrgantoBrainWeightPrinted:30-Mar-04PrincetonResearchCenterStudynumber:006130Page:1EastMillstone,NewJerseyVersion4.2.2Table13:MainStudy-TerminalSacrificeGroupTerminalBrainKidneysLungsBodywt.(g)AdrenalGlandsHeartLiverOvariesF e m a 1 e A n i m a 1 s1M e an:294.03.3732100.000052.0429116.3003409.499778.55854.5942Standarddeviation:24.50.42540.00004.97196.723340.35346.86920.8412Numberofobserv.:(10)(10)(10)(10)(10)(10)(10)(10)2M e an:288.53.7110100.000052.3417115.7209404.926077.91274.4751Standarddeviation:25.20.53460.00004.906415.058744.98919.57580.7969Numberofobserv.:(10)(10)(10)(10)(10)(10)(10)(10)3M e an:309.13.2164100.000053.2591126.2977436.490882.37224.3506Standarddeviation:32.00.73970.00005.269211.892175.69057.86790.7408Numberofobserv.:(10)(10)(10)(10)(10)(10)(10)(10)4M e an:284.63.3459100.000048.4561116.3956423.982877.15554.2777Standarddeviation:28.30.54340.00004.046111.125846.34709.04470.6114Numberofobserv.:(10)(10)(10)(10)(10)(10)(10)(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 227HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:MainStudy-TerminalSacrificeGroup1M e an:Standarddeviation:Numberofobserv.:2M e an:Standarddeviation:Numberofobserv.:3M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)294.024.5(10)288.525.2(10)309.132.0(10)284.628.3(10)SummaryStatisticsfor%OrgantoBrainWeightStudynumber:006130ThymusSpleenUterusFemaleA n i m a l s28.497015.787344.41483.22832.77919.7483(10)(10)(10)26.682616.121333.1895$4.54652.24832.7911(10)(10)(10)28.050116.164347.75004.64312.925017.8805(10)(10)(10)27.812814.350638.65114.02353.232313.0661(10)(10)(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 228Summarystatisticsforabsoluteorganweights(g)Studynumber:006130NHuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:TerminalSacrifice-NeurotoxBrainMeasurementGroup1M e an:Standarddeviation:Numberofobserv.:2M e an:Standarddeviation:Numberofobserv.:3M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)558.672.5(5)561.272.2(5)583.652.4(5)542.568.1(5)LengthweightWidthM a 1 eA n i m a 1 s1.68182.11.50.14820.00.0(5)(5)(5)1.75322.21.50.07100.10.0(5)(5)(5)1.78962.21.50.11890.10.0(5)(5)(5)1.75122.11.50.08760.10.0(5)(5)(5)Printed:30-Mar-04Page:1Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%{$)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 229Summarystatisticsforabsoluteorganweights(g)Studynumber:006130NHuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:TerminalSacrifice-NeurotoxBrainMeasurementsGroup1M e an:Standarddeviation:Numberofobserv.:2M e an:Standarddeviation:Numberofobserv.:3M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)330.540.3(5)318.524.5(5)315.953.5(5)316.729.6(5)LengthWeightWidthF e m a 1 eA n i m a 1 s1.64942.11.50.03400.00.0(5)(5)(5)1.69982.11.50.03090.00.0(5)(5)(5)1.67862.11.40.14980.00.1(5)(5)(5)1.70582.11.40.06780.00.0(5)(5)(5)Printed:30-Mar-04Page:1Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 230HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:RecoverySacrifice-MainStudyGroupSummarystatisticsforabsoluteorganweights(g)Studynumber:006130BrainHeartTerminalBodywt.(g)AdrenalGlandsEpididymisKidneysM a 1 e A n i m a 1 s1M e an:540.50.06862.24161.55431.69894.3624Standarddeviation:36.80.01070.10020.07510.17290.4534Numberofobserv.:(10)(10)(10)(10)(10)(10)4M e an:543.60.06912.26321.63571.72844.6263Standarddeviation:29.80.00890.09200.13610.14210.5443Numberofobserv.:(10)(10)(10)(10)(10)(10)Printed:30-Mar-04Page:1Version4.2.2LiverLungs14.86552.02321.29320.2499(10)(10)15.76602.09821.37370.1702(10)(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 231HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseySummarystatisticsforabsoluteorganweights(g)Studynumber:006130Table13:RecoverySacrifice-MainStudyGroup1M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)540.536.8(10)543.629.8(10)SeminalvesiclesProstateM a 1 e1.27881.84350.26420.5598(10)(10)1.18432.19060.18310.5179(10)(10)TestesSpleenA n i m a 1 s0.86873.42050.14580.2335(10)(10)0.89023.7421*0.18340.2829(10)(10)Thymus0.27510.0973(10)0.30010.0782(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 232HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:RecoverySacrifice-MainStudyGroupSummarystatisticsforabsoluteorganweights(g)Studynumber:006130BrainKidneysTerminalBodywt.(g)AdrenalGlandsHeartF e m a 1 e A n i m a 1 s1M e an:288.90.07462.14201.02822.3961Standarddeviation:22.20.01030.07640.06170.1492Numberofobserv.:(10)(10)(10)(10)(10)4M e an:294.50.07832.09811.18212.4379Standarddeviation:17.10.00650.05040.31980.1904Numberofobserv.:(10)(10)(10)(10)(10)Liver8.82770.5781(10)8.95861.2645(10)Printed:30-Mar-04Page:1Version4.2.2LungsOvaries1.53520.09200.09010.0255(10)(10)1.60760.09710.20250.0244(10)(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 233HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:RecoverySacrifice-MainStudyGroup1M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)288.922.2(10)294.517.1(10)Summarystatisticsforabsoluteorganweights(g)Studynumber:006130ThymusSpleenUterusF e m a 1 e A n i m a 1 s0.60310.25430.99480.07570.05860.2960(10)(10)(10)0.59870.2655 0.81990.09160.03990.3129(10)(10)(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 234HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:RecoverySacrifice-MainStudyGroupSummaryStatisticsfor%OrgantoBodyWeightStudynumber:006130BrainHeartTerminalBodywt.(g)AdrenalGlandsEpididymisM a 1 e A n i m a 1 s1M e an:540.50.01280.41590.28890.3144Standarddeviation:36.80.00240.02610.02560.0254Numberofobserv.:(10)(10)(10)(10)(10)4M e an:543.60.01280.41730.30200.3181Standarddeviation:29.80.00220.02550.03360.0225Numberofobserv.:(10)(10)(10)(10)(10)Kidneys0.80730.0661(10)0.85200.0977(10)Printed:30-Mar-04Page:1Version4.2.2LiverLungs2.75420.37650.21570.0588(10)(10)2.90150.38670.21550.0335(10)(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 235HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseySummaryStatisticsfor%OrgantoBodyWeightStudynumber:006130Table13:RecoverySacrifice-MainStudyGroup1M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt_(g)540.536.8(10)543.629.8(10)SeminalvesiclesProstateM a 1 e0.23900.33860.05930.0915(10)(10)0.21910.40340.04090.0946(10)(10)TestesSpleenA n i m a 1 s0.16030.63430.02220.0466(10)(10)0.16290.6898*0-02720.0600(10)(10)Thymus0.05070.0177(10)0.05500.0129(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%{$)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 236HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:RecoverySacrifice-MainStudyGroupSummaryStatisticsfor%OrgantoBodyWeightStudynumber:006130BrainKidneysTerminalBodywt.(g)AdrenalGlandsHeartF e m a 1 eA n i m a 1 s1M e an:288.90.02590.74510.35740.8320Standarddeviation:22.20.00420.05950.03020.0612Numberofobserv.:(10)(10)(10)(10)(10)4M e an:294.50.02660.71490.40160.8286Standarddeviation:17.10.00210.05070.10590.0585Numberofobserv.:(10)(10)(10)(10)(10)Liver3.07100.3050(10)3.03810.3681(10)Printed:30-Mar-04Page:1Version4.2.2LungsOvaries0.53490.03210.05760.0096(10)(10)0.54570.03290.06010.0076(10)(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 237HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:RecoverySacrifice-MainStudyGroup1M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)288.922.2(10)294.517.1(10)SummaryStatisticsfor%OrgantoBodyWeightStudynumber:006130ThymusSpleenUterusF e m a1e A n i m a 1 s0.20870.08840.34520.01970.02020.1004(10)(10)(10)0.20330.09010.27750.02800.01250.1000(10)(10)(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 238HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:RecoverySacrifice-MainStudyGroupSummaryStatisticsfor%OrgantoBrainWeightStudynumber:006130BrainHeartTerminalBodywt.(g)AdrenalGlandsEpididymisM a l eA n i m a l s1M e an:540.53.0693100.000069.511875.7644Standarddeviation:36.80.53710.00005.36186.6130Numberofobserv.:(10)(10)(10)(10)(10)4M e an:543.63.0600100.000072.4622 76.4156Standarddeviation:29.80.44300.00007.48116.1973Numberofobserv.:(10)(10)(10)(10)(10)Kidneys194.478316.4682(10)204.132419.2810(10)Printed:30-Mar-04Page:1Version4.2.2LiverLungs664.405390.574963.734313.7582(10)(10)696.840692.798955.86187.8662(10)(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 239HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseySummaryStatisticsfor%OrgantoBrainWeightStudynumber:006130Table13:RecoverySacrifice-MainStudyGroup1M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)540.536.8(10)543.629.8(10)SeminalvesiclesProstateM a 1 e57.201481.614412.079023.0587(10)(10)52.330697.33357.698324.8323(10)(10)TestesSpleenA n i m a 1 s38.8754152.66536.96949.1502(10)(10)39.3058165.7971*7.666116.6181(10)(10)Thymus12.34404.6503(10)13.23473.2593(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 240HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:RecoverySacrifice-MainStudyGroupSummaryStatisticsfor%OrgantoBrainWeightStudynumber:006130BrainKidneysTerminalBodywt.(g)AdrenalGlandsHeartFemaleA n i m a 1 s1M e an:288.93.4780100.000048.0365111.9848Standarddeviation:22.20.43270.00003.02778.0711Numberofobserv.:(10)(10)(10)(10)(10)4M e an:294.53.7344100.000056.2085116.2735Standarddeviation:17.10.32600.000014.19229.7910Numberofobserv.:(10)(10)(10)(10)(10)Liver412.208324.5428(10)426.618856.1668(10)Printed:30-Mar-04Page:1Version4.2.2LungsOvaries71.78544.30745.37231.2211(10)(10)76.66484.63349.72961.1826(10)(10)*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withDunnett'stestofsignificance%($)=meanvalueofgroupwassignificantlydifferentfromcontrolatP =0.05(0.01)withModifiedTtestofsignificancePage 241HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable13:RecoverySacrifice-MainStudyGroup1M e an:Standarddeviation:Numberofobserv.:4M e an:Standarddeviation:Numberofobserv.:TerminalBodywt.(g)288.922.2(10)294.517.1(10)SummaryStatisticsfor%OrgantoBrainWeightStudynumber:006130ThymusSpleenUterusF e m a 1 eA n i m a 1 s28.211211.913146.40723.91612.862013.5217(10)(10)(10)28.521812.668939.15124.17611.962915.1460(10)(10)(10)Printed:30-Mar-04Page:2Version4.2.2*(+)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withDunnett'stestofsignificance%{$)=meanvalueofgroupwassignificantlydifferentfromcontrolatP=0.05(0.01)withModifiedTtestofsignificancePage 242

# Huntingdon Life Sciences 00

- 6130 Page 243

211

- DIPE -S Final Report

Incidence Summary Report

for Gross Necropsy Observations

Preface Tab le 14

Table of Contents

Main Study

- Terminal Sacrifice

........................................................................ 244

Main Study

- Recovery Sacrifice

....................................................................... 245

Neuropathology Sacrifice

.................................................................................. 246

Key to Abbreviations

LN = Lymph Node

Submandib/Max = Submandibular/Submaxillary

Corresponding exposure levels for each group were as follow s:

Group 1

- 0 mg/m 3



# Group 2

- 2000 mg/m 3



# Group 3

- 10000 mg/m 3



# Group 4

- 20000 mg/m 3HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable14:MainStudy-TerminalSacrificeGroup:Numberingroup:Withinnormallimits..............EpididymisSmall•••••••••0•••••0••••••••••KidneysDilatedPelvis.................Cyst...........................LungsDiscolored.....................SeminalvesiclesSmall•0•••••00.000••0••••••••••Submandib/MaxLNDiscolored.....................Enlarged.......................TestesSmall••••00••••0••••••••000••••Soft...........................ThymusDiscolored.....................Skin-protocolHairThin/Absent0••••0.0••••••0110312120111000IncidenceSummaryReportforGrossNecropsyObservationsStudynumber:006130Males2 3910770100002201000001010100410501031000000IIIII110700010000002Females--2 310108 80000001100000000000011410900010000000Printed:15-Dec-11Page:1Version4.2.2Page 244HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyIncidenceSummaryReportforGrossNecropsyObservationsStudynumber:006130Table14:MainStudy-RecoverySacrificeGroup:Numberingroup:Withinnormallimits••••00••••••••LungsDiscolored•••••000•••••0.00••••ThymusDiscolored.....................Skin-protocolHairThin/Absent•0•••••••••••••VaginaNodule••0•••••••00•••00.0000.0.Males11071200410811001--1IIIIFemales--1 410108911001010Printed:15-Dec-11Page:1Version4.2.2Page 245HuntingdonLifeSciencesPrincetonResearchCenterEastMillstone,NewJerseyTable14:NeurotoxGroup:Numberingroup:Withinnormallimits•••0.0.0••0000LungsDiscolored••••0000•••••••••••0.KidneysDiscolored.....................ThymusDiscolored0••00••••••••••0000••Skin-protocolHairThin/Absent000•••000••••00Scab00.0••••••••••00•••••••00.015311000IncidenceSummaryReportforGrossNecropsyObservationsStudynumber:006130NMales2 35 55 5000000000045301111IIIII15500000Females2 35 5431 20000000045410000Printed:15-Dec-11Page:1Version4.2.2Page 246

# Huntingdon Life Sciences 00

- 6130 Page 247

211

- DIPE -S Final Report

Inc idence Summary of Microscopic

Findings with Severity Levels

Preface Table 15

Table of Contents

Main Study

- Terminal Sacrifice

........................................................................ 248

Main Study

- Recovery Sacrifice

....................................................................... 257

Neuropathology Sacrifice

.................................................................................. 264

Key to Abbreviations

Fibr = Fiber

Gang = Ganglia

LN = Lymph Node

Nad = No abnormal diagnoses

Nose/Turb Sec 1 = Nose/Turbinates Section 1

Nose/Turb Sec 2 = Nose/Turbinates Section 2

Nose/Turb Sec 3 = Nose/Turbinates Section 3

Nose/Turb Sec 4 = Nose/Turbinates Section 4

SC = Spinal Cord

Submandib/Max = Submandibular/Submaxillary

V- DVTC = Ventral Diverticulum

V- SM -G = Ventral Seromucinous G lands

Corresponding exposure levels for each group were as follows:

Group 1

- 0 mg/m 3



# Group 2

- 2000 mg/m 3



# Group 3

- 10000 mg/m 3



# Group 4

- 20000 mg/m 315-Dec-11;17:43HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--A nim a l sControlsfromgroup(s):1Animalsex:--M a l e s--Dosagegroup:Ctls2 3 4T i s s u e s W i t h D i agn o s e sNo.ingroup:1091010IAdrenalGlands.................................Numberexamined:100010ICONGESTIONNad>Minimal>............................TotalIncidenceofFindingObserved:Aorta..........................................Numberexamined:Brain..........................................Numberexamined:Cecum..........................................Numberexamined:CervicalSC....................................Numberexamined:Colon.................LUMEN:NEMATODE(S).........................Numberexamined:Nad>Present>............................TotalIncidenceofFindingObserved:Duodenum.......................................Numberexamined:Epididymis.....................................Numberexamined:Esophagus......................................Numberexamined:Femur..........................................Numberexamined:Heart..........................................Numberexamined:MONONUCLEARCELLINFILTRATEWITHMYOFIBERDEGENERATION/LOSSNad>Minimal>............................TotalIncidenceofFindingObserved:Ileum..........................................Numberexamined:Jejunum........................................Numberexamined:AllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeath10001010101010911101091010644101000000000000000000000000000000000000000000010001010101010100010101010109111010IIIrange:13-May-02To14-May-02A f fe c t e d--F e m a 1 e s--Ctls10109111010101010911101010108221010210000000000000000000000310000000000000000000000410101000101010101010001010101010001010Page 24815-Dec-11;17:43HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--Anim alsControlsfromgroup(s):1Animalsex:--M a l e s--IDosagegroup:Ctls2 3 4IT i s s u e sWi t h D i ag n o s e sNo.ingroup:1091010IKidneys........................................Numberexamined:100010ICORTEX:PROXIMALCONVOLUTEDTUBULAR-EPITHELIUM-EOSINOPHILIC/HYALINEDROPLETSNad>Minimal>Slight>............................TotalIncidenceofFindingObserved:BASOPHILICTUBULESNad>Minimal>Slight>............................TotalIncidenceofFindingObserved:CORTEX:LYMPHOIDCELLAGGREGATE(S)Nad>Minimal>............................TotalIncidenceofFindingObserved:CORTEX/CORTICO-MEDULLARYJUNCTION:MINERALDEPOSIT(S)Nad>Minimal>Slight>............................TotalIncidenceofFindingObserved:CORTEX/CORTICO-MEDULLARYJUNCTION:TUBULE(S)DILATEDNad>Minimal>............................TotalIncidenceofFindingObserved:10000721364410000911000000000000000000000000000000000000019103527288100001000AllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:13-May-02To14-May-02Af fe cted--Fe m ale s--Ctls2 3 41010101010100001000082245161000000000000000000000000000000000000000001010000820282264041000Page 24915-Dec-11;17:43HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--A nim a 1 sControlsfromgroup(s):1Animalsex:M a 1 e s--IDosagegroup:Ctls2 3 4IT i s s u e s W i t h D i a g n o s e sNo.ingroup:1091010IKidneys........................................Numberexamined:100010UROTHELIALHYPERPLASIANad>Minimal>............................TotalIncidenceofFindingObserved:PELVIS:DILATEDNad>Minimal>Slight>............................TotalIncidenceofFindingObserved:CORTICALCYSTNad>Present>............................TotalIncidenceofFindingObserved:Larynx:V-DVTC.................................Numberexamined:SUBMUCOSALLYMPHOCYTICAGGREGATENad>Minimal>............................TotalIncidenceofFindingObserved:SUBMUCOSALCYSTSWITHEOSINOPHILICCRYSTALSNad>Present>............................TotalIncidenceofFindingObserved:Larynx:V-SM-G.................................Numberexamined:GRANULOMATOUSCHONDROPATHYNad>Minimal>Slight>............................TotalIncidenceofFindingObserved:1000811291110555822101000000000000000000000000000000000000000000000000911910110001055510001010000AllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:13-May-02To14-May-02Af fe cted--Fe m a1e s--Ctls2 3 41010101010001010001000010001082273310712300000000000000000000000000000000000000000000100010000100010644911107303Page 25015-Dec-11;17:43HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--A n i m a 1 sControlsfromgroup(s):1Animalsex:--M a 1 e sIDosagegroup:Ctls234ITis s u e swi t h D i agn o s e sNo.ing:rou12:1091010ILarynx:V-SM-G.................................Numberexamined:100010ISUBMUCOSALLYMPHOCYTICAGGREGATENad>6008Minimal>4001Slight>0001............................TotalIncidenceofFindingObserved:4002Liver..........................................Numberexamined:100010MONONUCLEARCELLAGGREGATESNad>1002Minimal>9008............................TotalIncidenceofFindingObserved:9008LumbarSC......................................Numberexamined:100010ILungs..........................................Numberexamined:1091010ALVEOLAR/INTRAALVEOLARMACROPHAGESNad>1091010IMinimal>0000I............................TotalIncidenceofFindingObserved:0000IHEMORRHAGENad>9 9109Minimal>1001............................TotalIncidenceofFindingObserved:1001VISCERALPLEURA/CAPSULE:FIBROSISNad>1091010I............................TotalIncidenceofFindingObserved:0000IAllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:13-May-02To14-May-02A f fe c t e d--F e m a 1 e s--Ctls23 4101010101000106002400800004008100010400160096009100010101010101010108000200021010101000000000101010100000Page 25115-Dec-11;17:43HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--Anim alsControlsfromgroup(s):1Animalsex:M a l e sDosagegroup:Ctls2 3 4T i s s u e s W i t h D i a g n o s e sNo.ingroup:1091010IMediastinalLN.................................Numberexamined:9009ISINUSES:FREEERYTHROCYTESNad>Minimal>Slight>............................TotalIncidenceofFindingObserved:MesentericLN..................................Numberexamined:Nose/TurbSec1................................Numberexamined:ODONTOPATHYNad>Minimal>Moderate>............................TotalIncidenceofFindingObserved:SEPTALDEVIATIONNad>Present>............................TotalIncidenceofFindingObserved:NASOLACRIMALDUCT(S):LUMEN-HEMORRHAGE(S)Nad>Minimal>Slight>............................TotalIncidenceofFindingObserved:Nose/TurbSec2................................Numberexamined:SEPTALDEVIATIONNad>Present>............................TotalIncidenceofFindingObserved:351610101000010001000010100000000000000000000000000000000000000000000063031010901110009011101000AllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:13-May-02To14-May-02Af fe cted--F e m a l e s--Ctls2 3 41010101010730310109101100010000109110000000000000000000000000000000000000000000010721310109101911910110911Page 25215-Dec-11;17:43HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--Anim alsControlsfromgroup(s):1Animalsex:M a l e s--Dosagegroup:Ctls2 3 4T i s s u e s W i t h D i a g n o s e sNo.ingroup:1091010Nose/TurbSec2................................Numberexamined:100010ODONTOPATHYNad>Minimal>Slight>Moderate>............................TotalIncidenceofFindingObserved:NASOLACRIMALDUCT(S):LUMEN-EOSINOPHILICMATERIALNad>Slight>............................TotalIncidenceofFindingObserved:NASOLACRIMALDUCT(S):LUMEN-HEMORRHAGE(S)Nad>Minimal>Slight>............................TotalIncidenceofFindingObserved:Nose/TurbSec3................................Numberexamined:NASOLACRIMALDUCT(S):LUMEN-EOSINOPHILICMATERIALNad>Minimal>............................TotalIncidenceofFindingObserved:Nose/TurbSec4................................Numberexamined:Ovaries........................................Numberexamined:MINERALIZATIONNad>Minimal>............................TotalIncidenceofFindingObserved:Pancreas.......................................Numberexamined:Parathyroid....................................Numberexamined:1000008221000010911101010000000000000000000000000000000000000009010110009011101000101010AllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:13-May-02To14-May-02Af fe cted--Fe m ale s--Ctls2 3 410101010100010811021000100001010001010100010100000000000000000000000000000000000000000000000900111000910110100010109111010Page 25315-Dec-11;17:43HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--Ani m a 1 sControlsfromgroup(s):1Animalsex:--M a 1 e s--IDosagegroup:Ctls234IT i s s u e swi t hD i agn o s e sNo.ingrouQ:1091010IProstate.......................................Numberexamined:100010ILYMPHOIDCELLAGGREGATE(S)Nad>80010Minimal>2000............................TotalIncidenceofFindingObserved:2000Rectum/LowColon...............................Numberexamined:100010LUMEN:NEMATODE(S)Nad>7009IPresent>3001I............................TotalIncidenceofFindingObserved:3001ISalivaryglands................................Numberexamined:100010SciaticNerve..................................Numberexamined:100010ISeminalvesicles...............................Numberexamined:100110SECRETORYPRODUCT:DECREASEDNad>10009Minimal>0001Slight>0010............................TotalIncidenceofFindingObserved:0011Skin...........................................Numberexamined:0000IEPITHELIUM:SQUAMOUSCELLHYPERPLASIANad>0000IMinimal>0000I............................TotalIncidenceofFindingObserved:0000IEPITHELIUM:HYPERKERATOSISNad>0000Minimal>0000............................TotalIncidenceofFindingObserved:0000AllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:13-May-02To14-May-02A f fe c t e d--F e m a 1 e s--Ctls23 4101010101000109008100210021000101000102 1 10111010001000110010101010Page 25415-Dec-11;17>43HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--Anim a1sControlsfromgroup(s):1Animalsex:--M a 1 e s--IDosagegroup:Ctls2 3 4IT i s s u e s W i t h D i a g n o s e sNo.ingroup:1091010ISkin...........................................Numberexamined:0000IHYPOTRICHOSISMinimal>............................TotalIncidenceofFindingObserved:Spleen.........................................Numberexamined:Sternum........................................Numberexamined:Stomach........................................Numberexamined:GLANDULARECTASIANad>Minimal>............................TotalIncidenceofFindingObserved:Submandib/MaxLN...............................Numberexamined:SINUSES:FREEERYTHROCYTESNad>Minimal>Slight>............................TotalIncidenceofFindingObserved:Testes.........................................Numberexamined:SEMINIFEROUSTUBULES:DILATEDNad>Minimal>............................TotalIncidenceofFindingObserved:GERMINALEPITHELIUM:DEGENERATION/ATROPHYNad>Severe>............................TotalIncidenceofFindingObserved:ThoracicSC....................................Numberexamined:00101010100010811210911100010000000000000000000000000000000000011000110001010101000109011101000100010IIAllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:13-May-02To14-May-02Af fe cte d--Fe m a1e s--Ctls2 3 4101010102 1 10221010101000101000010110000000000001100000000000000101010911101000010Page 25515-Dec-11;17:43HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--Ani m a l sControlsfromgroup(s):1Animalsex:M a l e s--IDosagegroup:Ctls234IT i s s u e swi t hD i agn o s e sNo.ingrouj;:>:1091010IThymus.........................................Numberexamined:100110ICONGESTIONNad>9008Minimal>101 2............................TotalIncidenceofFindingObserved:101 2MEDULLARYHYPERPLASIANad>100110IMinimal>0000I............................TotalIncidenceofFindingObserved:0000IThyroidgland..................................Numberexamined:100010ITrachea........................................Numberexamined:100010IUrinarybladder................................Numberexamined:100010IUterus.........................................Numberexamined:ILUMEN:DILATEDNad>Slight>............................TotalIncidenceofFindingObserved:CERVIX:LYMPHANGIECTASIANad>IPresent>I............................TotalIncidenceofFindingObserved:IVagina.........................................Numberexamined:CYST............................TotalIncidenceofFindingObserved:IAllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:13-May-02To14-May-02A f fe c t e d--F e m a l e s--Ctls23 410101010100010100010000000009001010001000100010100010100010100010800920012001900101000100000000000Page 25615-Dec-11-17:44HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsRecoverySacrifice--Anim a1sControlsfromgroup(s):1Animalsex:--M a 1 e s--IDosagegroup:Ctls4IT i s s u e s W i t h D i a g n o s e sNo.ingroup:1010IAdrenalGlands.................................Numberexamined:00ICONGESTION............................TotalIncidenceofFindingObserved:Aorta..........................................Numberexamined:Brain..........................................Numberexamined:Cecum..........................................Numberexamined:CervicalSC....................................Numberexamined:Colon..........................................Numberexamined:LUMEN:NEMATODE(S)............................TotalIncidenceofFindingObserved:Duodenum.......................................Numberexamined:Epididymis.....................................Numberexamined:Esophagus......................................Numberexamined:Femur..........................................Numberexamined:Heart..........................................Numberexamined:MONONUCLEARCELLINFILTRATEWITHMYOFIBERDEGENERATION/LOSS............................TotalIncidenceofFindingObserved:Ileum..........................................Numberexamined:Jejunum........................................Numberexamined:AllDiagnoses;Phases:P3;Deathtypes:ScheduledFS;Dateofdeath000000000000000000000000000000range:14-Jun-02To14-Jun-02A f f e c t e dF e m a 1 e s--Ctls41010000000000000000000000000000000Page 25715-Dec-11-17:44HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsRecoverySacrifice--Anim a l sControlsfromgroup(s):1Animalsex:--M a l e sDosagegroup:Ctls4T i s s u e sWi t h D i ag n o s e sNo.ingroup:1010IKidneys........................................Numberexamined:00CORTEX:PROXIMALCONVOLUTEDTUBULAR-EPITHELIUM-EOSINOPHILIC/HYALINEDROPLETS............................TotalIncidenceofFindingObserved:BASOPHILICTUBULES............................TotalIncidenceofFindingObserved:CORTEX:LYMPHOIDCELLAGGREGATE(S)............................TotalIncidenceofFindingObserved:CORTEX/CORTICO-MEDULLARYJUNCTION:MINERALDEPOSIT(S)............................TotalIncidenceofFindingObserved:CORTEX/CORTICO-MEDULLARYJUNCTION:TUBULE(S)DILATED............................TotalIncidenceofFindingObserved:UROTHELIALHYPERPLASIA............................TotalIncidenceofFindingObserved:PELVIS:DILATED............................TotalIncidenceofFindingObserved:CORTICALCYST............................TotalIncidenceofFindingObserved:Larynx:V-DVTC.................................Numberexamined:SUBMUCOSALLYMPHOCYTICAGGREGATE............................TotalIncidenceofFindingObserved:SUBMUCOSALCYSTSWITHEOSINOPHILICCRYSTALS............................TotalIncidenceofFindingObserved:AllDiagnoses;Phases:P3;Deathtypes:ScheduledFS;Dateofdeath0000000000000000000000range:14-Jun-02To14-Jun-02A f f e c t e dF e m a l e s--Ctls41010000000000000000000000000Page 25815-Dec-11-17:44HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsRecoverySacrifice--Anim a1sControlsfromgroup(s):1Animalsex:--M a 1 e s--Dosagegroup:Ctls4T i s s u e s W i t h D i a g n o s e sNo.ingroup:1010Larynx:V-SM-G.................................Numberexamined:00GRANULOMATOUSCHONDROPATHY............................TotalIncidenceofFindingObserved:SUBMUCOSALLYMPHOCYTICAGGREGATE............................TotalIncidenceofFindingObserved:Liver..........................................Numberexamined:MONONUCLEARCELLAGGREGATES............................TotalIncidenceofFindingObserved:LumbarSC......................................Numberexamined:Lungs..........................................Numberexamined:ALVEOLAR/INTRAALVEOLARMACROPHAGESNad>............................TotalIncidenceofFindingObserved:HEMORRHAGENad>............................TotalIncidenceofFindingObserved:VISCERALPLEURA/CAPSULE:FIBROSISNad>Minimal>............................TotalIncidenceofFindingObserved:MediastinalLN.................................Numberexamined:SINUSES:FREEERYTHROCYTES............................TotalIncidenceofFindingObserved:MesentericLN..................................Numberexamined:AllDiagnoses;Phases:P3;Deathtypes:ScheduledFS;Dateofdeath00000000001 11 1001 100110000000000range:14-Jun-02To14-Jun-02A f f e c t e dF e m a 1 e s--Ctls410100000000000001111001100011010000000Page 25915-Dec-11-17:44HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsRecoverySacrifice--Anim a1sControlsfromgroup(s):1Animalsex:--M a 1 e s--IDosagegroup:Ctls4IT i s s u e s W i t h D i a g n o s e sNo.ingroup:1010INose/TurbSec1................................Numberexamined:00IODONTOPATHY............................TotalIncidenceofFindingObserved:SEPTALDEVIATION............................TotalIncidenceofFindingObserved:NASOLACRIMALDUCT(S):LUMEN-HEMORRHAGE(S)............................TotalIncidenceofFindingObserved:Nose/TurbSec2................................Numberexamined:SEPTALDEVIATION............................TotalIncidenceofFindingObserved:ODONTOPATHY............................TotalIncidenceofFindingObserved:NASOLACRIMALDUCT(S):LUMEN-EOSINOPHILICMATERIAL............................TotalIncidenceofFindingObserved:NASOLACRIMALDUCT(S):LUMEN-HEMORRHAGE(S)............................TotalIncidenceofFindingObserved:Nose/TurbSec3................................Numberexamined:NASOLACRIMALDUCT(S):LUMEN-EOSINOPHILICMATERIAL............................TotalIncidenceofFindingObserved:Nose/TurbSec4................................Numberexamined:Ovaries........................................Numberexamined:AllDiagnoses;Phases:P3;Deathtypes:ScheduledFS;Dateofdeathrange:000000000000000000000014-Jun-02To14-Jun-02A f f e c t e dF e m a 1 e s--Ctls4101000000000000000000000000000Page 26015-Dec-11-17:44HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsRecoverySacrifice--Anim a1sControlsfromgroup(s):1Animalsex:--Ma 1 e s--IDosagegroup:Ctls4IT i s s u e swithDiag n o s e sNo.ingroup:1010IOvaries........................................Numberexamined:MINERALIZATION............................TotalIncidenceofFindingObserved:Pancreas.......................................Numberexamined:Parathyroid....................................Numberexamined:Prostate.......................................Numberexamined:LYMPHOIDCELLAGGREGATE(S)............................TotalIncidenceofFindingObserved:Rectum/LowColon...............................Numberexamined:LUMEN:NEMATODE(S)............................TotalIncidenceofFindingObserved:Salivaryglands................................Numberexamined:SciaticNerve..................................Numberexamined:Seminalvesicles...............................Numberexamined:SECRETORYPRODUCT:DECREASED............................TotalIncidenceofFindingObserved:Skin...........................................Numberexamined:EPITHELIUM:SQUAMOUSCELLHYPERPLASIANad>............................TotalIncidenceofFindingObserved:EPITHELIUM:HYPERKERATOSISNad>............................TotalIncidenceofFindingObserved:AllDiagnoses;Phases:P3;Deathtypes:ScheduledFS;Dateofdeath000000000000000000000000000000range:14-Jun-02To14-Jun-02A f f e c t e dFe m a 1 e s--Ctls1000000000110104100000000000000Page 26115-Dec-11-17:44HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmuno.toxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsRecoverySacrifice--Anim a1sControlsfromgroup(s):1Animalsex:--M a 1 e sDosagegroup:Ctls4T i s s u e sWi t h D i ag n o s e sNo.ingroup:1010Skinexamined:00IMinimal>............................TotalIncidenceofFindingObserved:Spleen.........................................Numberexamined:Sternum........................................Numberexamined:Stomach........................................Numberexamined:GLANDULARECTASIA............................TotalIncidenceofFindingObserved:Submandib/MaxLN...............................Numberexamined:SINUSES:FREEERYTHROCYTES............................TotalIncidenceofFindingObserved:Testes.........................................Numberexamined:SEMINIFEROUSTUBULES:DILATED............................TotalIncidenceofFindingObserved:GERMINALEPITHELIUM:DEGENERATION/ATROPHY............................TotalIncidenceofFindingObserved:ThoracicSC....................................Numberexamined:Thymus.........................................Numberexamined:CONGESTIONMinimal>............................TotalIncidenceofFindingObserved:AllDiagnoses;Phases:P3;Deathtypes:ScheduledFS;Dateofdeath0000000000000000000000002 1212 1range:14-Jun-02To14-Jun-02A f f e c t e dF e m a 1 e s--Ctls1011100000000004100000000000000Page 26215-Dec-11-17:44HuntingdonLifeSciences006130MainStudyGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsRecoverySacrifice--Anim a1sControlsfromgroup(s):1Animalsex:--M a 1 e s--IDosagegroup:Ctls4IT i s s u e s W i t h D i a g n o s e sNo.ingroup:1010IThymus.........................................Numberexamined:2 1MEDULLARYHYPERPLASIANad>............................TotalIncidenceofFindingObserved:Thyroidgland..................................Numberexamined:Trachea........................................Numberexamined:Urinarybladder................................Numberexamined:Uterus.........................................Numberexamined:LUMEN:DILATED............................TotalIncidenceofFindingObserved:CERVIX:LYMPHANGIECTASIA............................TotalIncidenceofFindingObserved:Vagina.........................................Numberexamined:CYSTPresent>............................TotalIncidenceofFindingObserved:2000010000AllDiagnoses;Phases:P3;Deathtypes:ScheduledFS;Dateofdeathrange:14-Jun-02To14-Jun-02A f f e c t e dF e m a 1 e s--Ctls41010000000000000000000101010Page 26315-Dec-11;17:58HuntingdonLifeSciences006130NNeurotoxGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--A n i m a l sControlsfromgroup(s):1Animalsex:--M a l e s--IDosagegroup:Ctls2 34IT i s s u e swi t h D i agn o s e sNo.ingrou12:55 55IBrain..........................................Numberexamined:5005ICervicalSC....................................Numberexamined:5005DorsalRootFibr...............................Numberexamined:5005DorsalRootGang...............................Numberexamined:5005Eyes...........................................Numberexamined:5005CILIARYBODY:MONONUCLEARCELLINFILTRATENad>5005IMinimal>0000I............................TotalIncidenceofFindingObserved:0000IKidneys........................................Numberexamined:1001IPROXIMALTUBULAREPITHELIUM:EOSINOPHILICCRYSTALSNad>1000ISlight>000lI............................TotalIncidenceofFindingObserved:0001IDILATEDTUBULESNad>0001ISlight>1000I............................TotalIncidenceofFindingObserved:l000ILumbarSC......................................Numberexamined:5005ILungs..........................................Numberexamined:5 555IHEMORRHAGE(S)Nad>5 453IMinimal>0102I............................TotalIncidenceofFindingObserved:0102IAllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:16-May-02To17-May-02A f f e c t e d--F e m a l e sCtls23 45 5 5550055005500550055005400510001000000000000000000000000000000050055 5 55554500100010Page 26415-Dec-11;17:58HuntingdonLifeSciences006130NNeurotoxGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--Anim a1sControlsfromgroup(s):1Animalsex:--Male s--IDosagegroup:Ctls2 3 4IT i s s u e s W i t h D i aqn o s e sNo.inqroup:5 5 5 5ILungs..........................................Numberexamined:5 5 5 5ALVEOLARMACROPHAGESNad>Minimal>............................TotalIncidenceofFindingObserved:Opticnerve....................................Numberexamined:SciaticNerve..................................Numberexamined:Skin........examined:Marked>............................TotalIncidenceofFindingObserved:SURFACEBACTERIAPresent>............................TotalIncidenceofFindingObserved:DERMALGRANULATIONTISSUEModerate>............................TotalIncidenceofFindingObserved:NEUTROPHILICINFILTRATEModerate>............................TotalIncidenceofFindingObserved:HYPOTRICHOSISSlight>............................TotalIncidenceofFindingObserved:SuralNerve....................................Numberexamined:ThoracicSC....................................Numberexamined:500550000000000055500000000000000000411000000000000000500551111111111155AllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeathrange:16-May-02To17-May-02Af fe cted--F e m a 1 e sCtls2 35 5 5550055000000000004555000000000000000005500000000000000000455500550000000000055Page 26515-Dec-11;17:58HuntingdonLifeSciences006130NNeurotoxGasolineDIPEVaporCondensate:A13-WeekWhole-BodyInhalationToxicityStudyinRatswithNeurotoxicityAssessmentsand4-WeekInVivoGenotoxicityandImmunotoxicityAssessmentsIncidenceSummaryofMicroscopicFindingswithSeverityLevelsTerminalSacrifice--A nim a l sControlsfromgroup(s):1Animalsex:--M a l e s--Dosagegroup:Ctls2 3 4IT i s s u e s W i t h D i a g n o s e sNo.ingroup:5 5 5 5IThymus.........................................Numberexamined:0001CONGESTIONSlight>............................TotalIncidenceofFindingObserved:TibialNerve...................................Numberexamined:Trachea........................................Numberexamined:TrigeminalGang................................Numberexamined:VentrlRootFibr...............................Numberexamined:AllDiagnoses;Phases:P2;Deathtypes:ScheduledFS;Dateofdeath005555range:0010010055 5500500516-May-02To17-May-02A f f e c t e d--F e m a l e sCtls23 455 5 50000000000005005555 550055005Page 266

# Huntingdon Life Sciences 00

- 6130 Page 267

211

- DIPE

- S Final Report

Chamber Monitoring Results

Preface Append ix A

Key to Abbreviations

MMAD = Mass Median Aerodynamic Diameter

GSD = Geometric Standard Deviation

TMC = Total Mass ConcentrationHuntingdonLifeSciences00-6130DayDateExposureNominalNumberMean(mg/m3)(mg/m3)012-Feb-02100113-Feb-02200214-Feb-02300315-Feb-02400618-Feb-02500719-Feb-02600820-Feb-02700921-Feb-028001022-Feb-029001325-Feb-0210001426-Feb-0211001527-Feb-0212001628-Feb-0213001701-Mar-0214002004-Mar-0215002105-Mar-0216002206-Mar-0217002307-Mar-0218002408-Mar-0219002711-Mar-0220002812-Mar-0221002913-Mar-0222003014-Mar-022300GASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGrouplA-0mg/m3(AirControl)AnalyticalChamberConcentrationIndividualMMAD(mg/m3)00000000000000001.2090000000000000000000000004.471000000000000000000002.303000000000000000000001.520000000000000ParticleSizeDeterminationsGSDTMC(mg/m3)1.4951.94E-032.6827.37E-032.3693.03E-031.8763.20E-03--ChamberEnvironmentMeanTemperatureHumidity(OC)(%)24572558255825562556245824542454245524512658255325572558255424442454245924542455255221532554Page 268HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)3115-Mar-022403216-Mar-022503418-Mar-022603519-Mar-022703620-Mar-022803721-Mar-022903822-Mar-023004125-Mar-023104226-Mar-023204327-Mar-023304428-Mar-023404529-Mar-02350481-Apr-02360492-Apr-02370503-Apr-02380514-Apr-02390525-Apr-02400558-Apr-02410569-Apr-024205710-Apr-024305811-Apr-024405912-Apr-024506215-Apr-024606316-Apr-02470MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGrouplA-0mg/m3(AirControl)AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(.urn)00000000000.90530000000000000000000000000000002.495000000000000000000002.35700000000000000000000000002.18700000000000000000000000001.4130000000000ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)25532.1371.20E-022754255324542452245324582.1304.91E-0324532551255225552.5017.60E-03255425502456235224552.0492.32E-03235524512551255324521.9988.01E-03245025472454Page 269HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)6417-Apr-024806518-Apr-024906619-Apr-025006922-Apr-025107023-Apr-025207124-Apr-025307225-Apr-025407326-Apr-025507629-Apr-025607730-Apr-02570781-May-02580792-May-02590803-May-02600836-May-02610847-May-02620858-May-02630869-May-026408710-May-026508912-May-026609013-May-026709114-May-026809215-May-026909316-May-02700Mean0S.D.0MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGrouplA-0mg/m3(AirControl)AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(pm)0000000000000001.76700000000000000000000000002.13000000000000000000000000001.32500000000000000000000000001.594000000000000000000000000001.97500.899ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)245224522.8585.16E-03244924502554255525521.9274.07E-03255425492453245324511.6051.29E-03245424512452255124512.0932.01E-032450255024542253225622542.1324.84E-0324.353.30.3923.13E-030.92.8Page 270HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)012-Feb-0210113-Feb-0220214-Feb-0230315-Feb-0240618-Feb-0250719-Feb-0260820-Feb-0270921-Feb-02801022-Feb-02901325-Feb-021001426-Feb-021101527-Feb-021201628-Feb-02130171-Mar-02140204-Mar-02150215-Mar-02160226-Mar-02170237-Mar-02180248-Mar-021902711-Mar-022002812-Mar-022102913-Mar-022203014-Mar-022303115-Mar-02240MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIB-0mg/m3(AirControl)AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(.urn)000000000000000000001.2300000000000000000000000000000003.22200000000000000000000000002.09000000000000000000000000001.46500000000000000000000Chamber EnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)2358235924601.6271.72E-032458245724592455245424572.3097.63E-03245425592456236924581.8073.69E-03245524452455255924551.9662.72E-0324552355225624572355Page 271HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)3216-Mar-022503418-Mar-022603519-Mar-022703620-Mar-022803721-Mar-022903822-Mar-023004125-Mar-023104226-Mar-023204327-Mar-023304428-Mar-023404529-Mar-02350481-Apr-02360492-Apr-02370503-Apr-02380514-Apr-02390525-Apr-02400558-Apr-02410569-Apr-024205710-Apr-024305811-Apr-024405912-Apr-024506215-Apr-024606316-Apr-024706417-Apr-02480MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroup18-0mg/m3(AirControl)AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(pm)000000.99330000000000000000000000000000003.236000000000000000000001.24000000000000000000000000002.02300000000000000000000000001.877000000000000000ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)2.5591.41E-022653245124552453245324582.5793.65E-0324542355245424582.3744.86E-03245624522456245324561.9942.81E-03245624522456245523552.9329.96E-032353234924552454Page 272HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)6518-Apr-024906619-Apr-025006922-Apr-025107023-Apr-025207124-Apr-025307225-Apr-025407326-Apr-025507629-Apr-025607730-Apr-02570781-May-02580792-May-02590803-May-02600836-May-02610847-May-02620858-May-02630869-May-026408710-May-026508912-May-026609013-May-026709114-May-02680Mean0S.D.0MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIB-0mg/m3(AirControl)AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(.urn)00000000001.879000000000000000000000000011.6200000000000000000000000001.50300000000000000000000000001.70800000000000000002.62202.790ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)24532.4605.66E-03245024512358245823552.6431.06E-02235324522454245424531.7591.54E-03255525512355245423521.7492.43E-0323542353235722542.2125.49E-0323.855.00.4173.94E-030.73.1Page 273HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)012-Feb-0211940113-Feb-0221990214-Feb-0231950315-Feb-0242150618-Feb-0252010719-Feb-0262090820-Feb-0272060921-Feb-02821301022-Feb-02920001325-Feb-021020901426-Feb-021119701527-Feb-021219901628-Feb-02132030171-Mar-02142030204-Mar-02152050215-Mar-02161900226-Mar-02172100237-Mar-02182090248-Mar-021921502711-Mar-022020202812-Mar-022120102913-Mar-022220103014-Mar-022320803115-Mar-02242160MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIIA-2,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(pm)198519901860202020701930177019102020202019532000199020201800208519002170215021201.4221995198020001980202020251920201020702100198519201980202020201998200019402030202019502300161019401950199519201900205021101.18219982080191020002000195819201960201019402010205019801990202020352040205020502000199820901980200019201.83218651940173018001990200020101990200020001915202017501970192019831900203020201980192820201820197019001.50618831850213017901760198819502020198020001905186018401900202019681870192019502130ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(oC)(%)2352245323521.9333.09E-032450245024522448244824492.0604.38E-03244825502448235123522.3212.65E-03245023422350245523502.0533.51E-0324522350224923502350Page 274HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)3216-Mar-022521903418-Mar-022621903519-Mar-022721603620-Mar-022821303721-Mar-022921803822-Mar-023020504125-Mar-023120904226-Mar-023220304327-Mar-023321704428-Mar-023421104529-Mar-02352110481-Apr-02362090492-Apr-02372030503-Apr-02382130514-Apr-02392060525-Apr-02402220558-Apr-02411980569-Apr-024220405710-Apr-024320505811-Apr-024420605912-Apr-024520606215-Apr-024620606316-Apr-024719106417-Apr-02482000MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIIA-2,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(pm)206819002050210022200.79402205233021902280202020952100206021102110204020202030207020402108217020402110211019131950194018401920191819001900183020403.911191320801760183019802133222021302150203020902130203020202180202820402040201020200.842218432120163018301790184519001790181018801875179018901890193020932240213020101990215018602030234023701.43819502010181017302250193819802030172020201955201019501920194019602010203019601840182018701890173017901.154195820102110173019802040222020201920200020802240202020102050ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)1.9081.19E-022650234823502349I2350I2354I2.4464.72E-0323512448244824522.224 4.56E-03245024472352234823512.0531.53E-03235123482447244923492.0455.62E-032348234523512348Page 275HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)6518-Apr-024920106619-Apr-025021406922-Apr-025119307023-Apr-025219707124-Apr-025320607225-Apr-025419707326-Apr-025520607629-Apr-025620607730-Apr-02571900781-May-02582090792-May-02592040803-May-02602070836-May-02612060847-May-02622120858-May-02632190869-May-026419908710-May-026520808912-May-026621309013-May-026722309114-May-026823309215-May-026921509316-May-02702100Mean2068S.D.83MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIIA-2,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3){tim)20002040202019202020210820302130218020901.072188517001970202018501963201018702020195020451950239018801960187519201970173018802020201020202040201012.3419852030197020201920189818801930192018602163229022102110204020652150212019802010201020201990200020301.35119582150200018501830193519201950191019601890188020101750192018081880179018301730191521702020175017201.631194320101660182022801945192019501970194020152070201019702010202321601900201020201883190018901790195019812.3441293.103-ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)23491.8033.00E-03234623472450245124482.8756.96E-03244924462349II2349I23471.7831.34E-03244923472350234923461.8702.36E-032348234822512251225222492.1064.28E-0323.449.40.3032.79E-030.72.1----Page 276HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)012-Feb-0211940113-Feb-0221990214-Feb-0231950315-Feb-0242150618-Feb-0252010719-Feb-0262090820-Feb-0272060921-Feb-02821301022-Feb-02920001325-Feb-021020901426-Feb-021119701527-Feb-021219901628-Feb-02132030171-Mar-02142030204-Mar-02152050215-Mar-02161900226-Mar-02172100237-Mar-02182090248-Mar-021921502711-Mar-022020202812-Mar-022120102913-Mar-022220103014-Mar-022320803115-Mar-02242160MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupliB-2,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(.um)196321001810198019601943206019501900186019832020192018802110217824002030213021501.5892135201021702210215020632180199020002080194319801930192019402035209021201980195019031910171020001990207021602170196019901.37819381730192020002100199319202000201020401975206020002000184020501920215022101920211322102080204021203.38019182000186018601950199820101980200020002028219018902020201019581910202019501950202317601960218021901.1741963208022101850171019732040200019201930198519601950194020902020192019802160ParticleSizeDeterminationsGSDTMC(mg/m3)2.3213.46E-032.2735.03E-032.8273.14E-031.6332.54E-03----ChamberEnvironmentMeanTemperatureHumidity(oC)(%)235423562356235524542355235123502351234924542352235523552352224322522356225123532352225223532352Page 277HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)3216-Mar-022521903418-Mar-022621903519-Mar-022721603620-Mar-022821303721-Mar-022921803822-Mar-023020504125-Mar-023120904226-Mar-023220304327-Mar-023321704428-Mar-023421104529-Mar-02352110481-Apr-02362090492-Apr-02372030503-Apr-02382130514-Apr-02392060525-Apr-02402220558-Apr-02411980569-Apr-024220405710-Apr-024320505811-Apr-024420605912-Apr-024520606215-Apr-024620606316-Apr-02471910_6417-Apr-02482000MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroup118-2,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3){tim)209819502200219020500.89982020208020101980201020282150193020102020204021102060198020102073224020202020201019602040199018901920196819802050208017600.9819196016402000222019801978203019801950195019152020192018901830199820201980196020300.889721301810240021802130196021301840185020201970190020202040192020982170222020201980205522701890211019501.40318581790179017302120186817701780215017701943179019801980202019852100202019601860197020502020183019801.101200019302180179021001903214018401790184019202050192017301980-----·ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)2.471.47E-022552235023522351235123551.7621.76E-03235223522350I23541.9265.04E-03235323482354234923541.9891.74E-03235423492352235222521.7556.24E-032250224723532350Page 278HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)6518-Apr-024920106619-Apr-025021406922-Apr-025119307023-Apr-025219707124-Apr-025320607225-Apr-025419707326-Apr-025520607629-Apr-025620607730-Apr-02571900781-May-02582090792-May-02592040803-May-02602070836-May-02612060847-May-02622120858-May-02632190869-May-026419908710-May-026520808912-May-026621309013-May-026722309114-May-02682330Mean2067S.D.83MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupliB-2,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3){tim)19131860215017201920213322502180205020500.918720552300196020501910204322402040202018702133209024001990205020832310218019201920216822802130218020801.35820852100206021102070182822101730170016702068203020502090210019832000187020202040208321102180202020201.31420481940200021202130199520202000198019801973200021701740198020151920193020902120196322502040177017901.51020432030194019302270209521302090211020502075216021002020202020091.3771340.647ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)22521.5582.74E-03234823482353235522521.6922.33E-03235123492352235123501.6311.16E-03235323482253235122501.7202.23E-0322502250225322531.9664.01E-0322.851.80.3903.54E-030.52.4Page 279HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)012-Feb-0219470113-Feb-0229450214-Feb-0239540315-Feb-0249890618-Feb-0259830719-Feb-0269740820-Feb-02710800921-Feb-02899101022-Feb-02999501325-Feb-021098401426-Feb-021199901527-Feb-021299201628-Feb-021310100171-Mar-021410100204-Mar-02159200215-Mar-02169890226-Mar-02179730237-Mar-02189860248-Mar-021999602711-Mar-022098102812-Mar-022198902913-Mar-022297003014-Mar-0223101003115-Mar-02249980MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGrouplilA-10,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(tim)9685100009580958095809850929010200991010000929596508630935095501040099101150010200100000.98891031010600104009940103009925101009650965010300968099109190942010200103809910106001060010400975810200773010200109001048011400102009910104001.0439823994010500991089409943106009420991098401065010700103001070010900107001120010500102001090099351120088401060091001.708102009580106001050010100104801020010700107001030010420100001110010700987010240965010100108001040010480108001020010700102002.2451028099101080010200102009968109009580981095801003083001030011500100009973102009580991010200ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)2447244824471.8523.59E-032447244724502449245124502.1515.74E-03244725492447244724472.1832.65E-03244623392347245123502.6224.77E-0324472347224924502350Page 280HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)3216-Mar-022595303418-Mar-0226100003519-Mar-0227102003620-Mar-0228101003721-Mar-02"29103003822-Mar-023093704125-Mar-023196904226-Mar-023299404327-Mar-023393904428-Mar-023495404529-Mar-02359670481-Apr-02369830492-Apr-023710000503-Apr-023810600514-Apr-023910000525-Apr-02409470558-Apr-024110000569-Apr-024298805710-Apr-0243101005811-Apr-024499605912-Apr-024598406215-Apr-024699506316-Apr-024799506417-Apr-02489460MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGrouplilA-10,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(pm)1026010800106001190077200.7927100001000010100991010000103201080097401110096501001098401040095801020099709580100001030010000947594201030084409740945097409350952091901.31697731070091909910929096701000099109420935099308620100001050010600100109580104009870102000.77209790984097409580100009970100009840974010300973093201030097809520974510200952097409520909891008550893098101.490104501030010400106001050098281060010000929094209945978010000100001000010730109001070010500108009475926082601060097800.9063974310000105008170103001059010700103001170096501083011200102001120010700ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)2.4522.41E-022650244723502350234923502.4512.91E-0323482448244824481.8895.54E-03244924472350234823482.2491.78E-03234923472449244924491.6627.08E-032448244623522350Page 281HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)6518-Apr-0249101006619-Apr-025094706922-Apr-025195507023-Apr-025297807124-Apr-025397807225-Apr-025499407326-Apr-025596807629-Apr-025696207730-Apr-02579680781-May-025810200792-May-02599730803-May-026010100836-May-026110000847-May-02629670858-May-02639460869-May-026496908710-May-026593008912-May-026698609013-May-026798109114-May-026897109215-May-026995009316-May-02709390Mean9821S.D.286MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGrouplilA-10,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(tim)10090101001050099109840963399109260968096801.6181028010200117009100101001110010700108001090012000923391308470968096509770991010100942096501009010100101001090092601.2859488906088109580105009468987093501020084501001097401010010100101009400968099109290872010380102001000011000103001.24398401000098101000095509318100009650919084309658913084001080010300924087409610877098409490942095208620104004.537994096801020095801030096931070092609260955093658740929010200923010790965010400117001140010210100009740109001020099731.5347010.992ChamberEnvironmentiParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)23502.0066.01E-03234623472447244724471.7011.84E-03244724452348244823481.6461.17E-03244724472350244923472.5713.25E-032348234823512249224922492.1105.42E-0323.548.20.3475.91E-030.71.8-----Page 282HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)012-Feb-0219470113-Feb-0229450214-Feb-0239540315-Feb-0249890618-Feb-0259830719-Feb-0269740820-Feb-02710800921-Feb-02899101022-Feb-02999501325-Feb-021098401426-Feb-021199901527-Feb-021299201628-Feb-021310100171-Mar-021410100204-Mar-02159200215-Mar-02169890226-Mar-02179730237-Mar-02189860248-Mar-021999602711-Mar-022098102812-Mar-022198902913-Mar-022297003014-Mar-0223101003115-Mar-02249980MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIIIB-10,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(.urn)993010200974095801020099251040098409550991010650105001060010800107001027010900916010400106001.3519998104009710958010300100101010097409780104001015097801000010000108001015010700100009910100001024011000844011300102009480846095809480104001.01310120948099101050010600107409940103001120011500106501080010800107001030010010102001010095201020099559580994010000103001.7291014010900103001010092609743101001000095209350101709350108001060099109590910094801020095809655102009580942094201.44399951000010200991098709168814010400910090301010010000958010600102009983103009420991010300----ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)23462346I24472.3644.54E-032447I244624462446244724452.2076.20E-03244525492446244724472.3392.53E-03244623372344244723461.8263.34E-0323442445234823492349-Page 283HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)3216-Mar-022595303418-Mar-0226100003519-Mar-0227102003620-Mar-0228101003721-Mar-0229103003822-Mar-023093704125-Mar-023196904226-Mar-023299404327-Mar-023393904428-Mar-023495404529-Mar-02359670481-Apr-02369830492-Apr-023710000503-Apr-023810600514-Apr-023910000525-Apr-02409470558-Apr-024110000569-Apr-024298805710-Apr-0243101005811-Apr-024499605912-Apr-024598406215-Apr-024699506316-Apr-024799506417-Apr-02489460MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroup1118-10,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3){tim)9325958095201020080000.786699801020010300968097409548935091009740100009510916096509230100001011095201000010300106009188965010000855085509573965093501000092904.1911043011100104001000010200981810200984095809650949395809230958095801007010900101009190101000.725910260105001010099401050090338100935089409740106309910115001040010700106801100010700108001020010250104001020010400100001.7679763991098109910942010130106009940103009680107801030011100106001110092039520836093509580977510400850010200100000.8941988897801070092309840103101060096501100010000102401060097401040010200ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)2.3302.44E-022649234624462446244624463.2035.14E-032445!2346234623461.6335.49E-03244724442447244624452.1943.29E-03244524452448234623471.8378.92E-032346234424472446Page 284HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)6518-Apr-0249101006619-Apr-025094706922-Apr-025195507023-Apr-025297807124-Apr-025397807225-Apr-025499407326-Apr-025596807629-Apr-025696207730-Apr-02579680781-May-025810200792-May-02599730803-May-026010100836-May-026110000847-May-02629670858-May-02639460869-May-026496908710-May-026593008912-May-026698609013-May-026798109114-May-02689710Mean9832S.D.282MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroup1118-10,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(pm)1100011500113001060010600100709940974010400102000.8013100901020010400945010300975891309480932011100106501070010200110001070010240974010200105001050010530101001040011200104002.01398859840942095801070011250114001110011400111001113010900115001100011100104801080010100106001040010650104001020011600104001.66111180113001110011100112001035010600107001040097109923929094801160093201050010200105001000011300101801040094209910110001.7361055011500981010100108009650102001010086509650921887108580100009580100901.5476820.909ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(oC)(%)24471.9047.49E-03244424442446244623472.5465.59E-032446244424452445'24452.3323.48E-03254425442347234723451.9672.83E-0323462346224923462.2066.40E-0323.745.90.4025.73E-030.71.7Page 285HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)012-Feb-02118400113-Feb-02217700214-Feb-02318400315-Feb-024205006 '18-Feb-02520000719-Feb-02619600820-Feb-02720000921-Feb-028197001022-Feb-029196001325-Feb-0210196001426-Feb-0211199001527-Feb-0212194001628-Feb-021319100171-Mar-021418900204-Mar-021520000215-Mar-021619600226-Mar-021718500237-Mar-021819200248-Mar-0219190002711-Mar-0220188002812-Mar-0221195002913-Mar-0222193003014-Mar-0223191003115-Mar-022419100MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIVA-20,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(tim)1968019400 1920019700204001988020100189001930021200194802010018700192001990020430 203002030020700204001.77920600197002220020500200002013019800203001930021100206502090020900207002010020550 20300 2030020500 2110021400 2120022300 210002110020480 21600 2040020400195003.1982050020000 20800204002080020600197002070020900211002003020700 2060020700181002030020400 2040020100 203002075020800 2090020800205001.192209502090021300208002080020500211002010020400 204001990019500201002010019900200501980020500199002000019680189001810021000207001.35820130 20300 20100201002000020950 212002130020900204001938018200 1950020500193001910018200185002010019600ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)2548255025502.6304.97E-032551254925502450245324513.8121.15E-02245026532550255025511.7791.83E-03254924402449255224522.1704.54E-0325482549225125512552Page 286HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)3216-Mar-0225194003418-Mar-0226190003519-Mar-0227193003620-Mar-02281980037'21-Mar-0229190003822-Mar-0230195004125-Mar-0231193004226-Mar-0232193004327-Mar-0233196004428-Mar-0234184004529-Mar-023518600481-Apr-023618700492-Apr-023718900503-Apr-023818800514-Apr-023918900525-Apr-024019100558-Apr-024118800569-Apr-0242176005710-Apr-0243197005811-Apr-0244184005912-Apr-0245194006215-Apr-0246187006316-Apr-0247189006417-Apr-0248187006518-Apr-024918800MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIVA-20,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(tim)19980196002050020100197000.77391988018900203001980020500192501950019000195001900019650182002040019900201001935019600195001900019300202801990020300204002050019550195001970020100189002.34620330204002010020700201002025019800197002100020500197801950019800202001960019480194001890020300193000.8053200802010019700200002050020450208002020020300205002030020100198002040020900194801930019000200001960020100197001950020800204001.220198301990019500193002060019050189001880019000195002078020200208002130020800203302050020500201002020020680204002110021100201001.3732058020200204002040021300209002080021000210002080020100204002000020700193002065020400208002040021000ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)1.9492.44E-022852254824512450255025513.1914.30E-0325492549254925482.0047.67E-03264926472550245224501.5491.09E-03245025492550265125493.0111.12E-0225492548245424512451Page 287HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)6619-Apr-0250184006922-Apr-0251186007023-Apr-0252189007124-Apr-0253187007225-Apr-0254190007326-Apr-0255188007629-Apr-0256190007730-Apr-025719000781-May-025818900792-May-025918800803-May-026018800836-May-026119000847-May-026219100858-May-026319100869-May-0264186008710-May-0265189008912-May-0266191009013-May-0267195009114-May-0268191009215-May-0269187009316-May-027019300Mean19070S.D.510MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIVA-20,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(pm)20430205002110019700204000.9267203301980020500201002090021200212002110021200213002048019800203002060021200206302050021700201002020021080212002010022200208001.413198301930020300196002010020800206002120020200212002035020400202002010020700202302060020300203001970020100209001970019900199003.580209302030020900216002090020100207001900020000207002013020700200002040019400185501720019100189001900019180189001870019800193001.59718830183001800019300197001958019000193001960020400194502000019800193001870019900200002010019500200002030020200203002030020400201401.659770_____!!;878ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(oC)(%)1.5782.63E-03244824482548254925491.8261.91E-03254926472449255024502.8636.05E-03254925482552255125481.6811.85E-032449254924522349235123512.3116.46E-0324.749.70.7196.38E-030.81.9Page 288HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)012-Feb-02118400113-Feb-02217700214-Feb-02318400315-Feb-02420500618-Feb-02520000719-Feb-02619600820-Feb-02720000921-Feb-028197001022-Feb-029196001325-Feb-0210196001426-Feb-0211199001527-Feb-0212194001628-Feb-021319100171-Mar-021418900204-Mar-021520000215-Mar-021619600226-Mar-021718500237-Mar-021819200248-Mar-0219190002711-Mar-0220188002812-Mar-0221195002913-Mar-0222193003014-Mar-0223191003115-Mar-022419100MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIVB-20,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(pm)20000217002010018800194001920018200192001930020100199001980021100179002080022680233002260022600222001.1342168022500209002250020800213001920022000214002260020400200002070020200207002003019600196002010020800205302040021400202002010021200205002130021500215001.041213802170022600217001950020530209001990020500208002045020700205002020020400209002130021700202002040021100207002120021100214001.147214502140021700220002070019950200001970020000201002100021700207002090020700196501940019900195001980019350193001740020700200001.1822035020700205002000020200194001940019500192001950021080207002070021200217002043020800196002060020700IIChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3j(OC)(%)'2449245024511.9174.00E-03245224502549255025522553I2.1117.73E-0325502553I24512451!2450!1.7821.69E-0324502540I2548I25512551I2.0564.70E-032547I24482351I2451I2451Page 289HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)3216-Mar-0225194003418-Mar-0226190003519-Mar-0227193003620-Mar-0228198003721-Mar-0229190003822-Mar-0230195004125-Mar-0231193004226-Mar-0232193004327-Mar-0233196004428-Mar-0234184004529-Mar-023518600481-Apr-023618700492-Apr-023718900503-Apr-023818800514-Apr-023918900525-Apr-024019100558-Apr-024118800569-Apr-0242176005710-Apr-0243197005811-Apr-0244184005912-Apr-0245194006215-Apr-0246187006316-Apr-0247189006417-Apr-024818700MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIVB-20,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(.um)20680203002120020500207000.75782098020700213002030021600205802070020400207002050020800205002080020800211002073020300206002110020900207502070020700211002050021030210002120020700212000.901819850200002000019900195002038020000203002080020400199301890020400204002000019650194002080019900185000.7779189301850018900192001910018830179001900019100193001925019700189001890019500199001960019000207002030019700196002060019300193001.517198801960019300200002060018230183001820018100183002035020100208002080019700201502050020100200002000020200198002080020200200001.301196501960019200198002000020450201002080020800201001930018100192002020019700ChamberEnvironmentParticleSizeMeanDeterminationsTemperatureHumidityGSDTMC(mg/m3)(OC)(%)1.8422.31E-022652244925502549254925502.1953.14E-0325482451245024502.1149.04E-03245024482549255025492.0382.38E-03254825482552245024492.2539.39E-032449244825532551Page 290HuntingdonLifeSciences00-6130DayDateExposureNominalNumber(mg/m3)6518-Apr-0249188006619-Apr-0250184006922-Apr-0251186007023-Apr-0252189007124-Apr-0253187007225-Apr-0254190007326-Apr-0255188007629-Apr-0256190007730-Apr-025719000781-May-025818900792-May-025918800803-May-026018800836-May-026119000847-May-026219100858-May-026319100869-May-0264186008710-May-0265189008912-May-0266191009013-May-0267195009114-May-026819100Mean19070S.D.515MeanGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSChamberMonitoringResultsCumulativeExposureRecordGroupIVB-20,000mg/m3AnalyticalChamberConcentrationIndividualMMAD(mg/m3)(mg/m3)(tim)200802040020400197001980018850185001940018300192000.7490197801900020000194002070019850205001990020500185001943019200198001900019700194801940020400196001850019930208001920019200205001.357193001880019400193001970019030184001920018400201001990020200201001930020000188001790019200194001870019200190001950019300190006.591197501950020400197001940019680204001860019700200002033020900202002070019500200002020020100197002000020000200002010019900200001.699203502000020000208002060020100200002000019800206001888020000190001830018200201001.550ParticleSizeDeterminationsGSDTMC(mg/m3)1.5935.03E-031.9973.45E-033.0863.95E-032.0252.97E-032.0786.20E·03967-1.§42-0.3515.64E-03-----------ChamberEnvironmentMeanTemperatureHumidity(aC)(%)'2551254825482451245024502449244825492549254926492649245224512449244924492454234924.549.80.61.9Page 291

# Huntingdon Life Sciences 00

- 6130 Page 292

211

- DIPE

- S Final Report

Individual Clinical Observations

Preface Appendix B

For summarization purposes, descriptive comments [i.e., location of scab(s)

and/or sore(s), etc.] are not presented in this table. These data are contained in

the study raw data if needed.

Corresponding exposure levels for each group were as follows:

Group I

– 0 mg/m 3



# Group II

– 2000 mg/m 3



# Group III

– 10000 mg/m 3



# Group IV

– 20000 mg/m 325-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)MALESGROUP10MG/M3APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 -1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 2ANIMAL#1051105210531054105510561057105810591060OBSERVATIONSWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEDERMALGENERAL:ALOPECIA-EXTREMITIES/SNOUTWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEOCULAR:CHROMODACRYORRHEA-UNILATERALCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENT1 1 6 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p p p p p p p ppp p p p p p p p p p p p p p p p p p ppp p p p p p p p p p p p p p pp p p ppp p p p p p p p p p p p p p p p p p ppp p p p p p p p p p p p p p p p p p ppp p p p p p p p p p p p p p p p p p ppp p p p p p p p p p p p p p pp p p ppp p p p pp p pp3 3 3 3 3 3 3 3 3 33p p p p p p p p p p p p p p p p p p ppp p p p p p p p p p p pp pp p p pppPage 29325-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSMALESANIMAL#10611062106310641065106610671068GROUP10MG/M3OBSERVATIONSDAYOFSTUDYWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEOCULAR:CHROMODACRYORRHEA-UNILATERALWITHINNORMALLIMITSTERMINALSACRIFICEDERMALGENERAL:SCABSOCULAR:CHROMODACRYORRHEA-UNILATERALORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEDERMALGENERAL:SCABSCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENT1 1 1 11 -1 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 1 6307 4 1 8 52 9 6 301 7 4 1 82p p p p p p p p p p p pp p p p p pp pp pp pp pp p p p p p p pppp p p p p p pp p p p p p p p p p p p p pp p p p p p p p p p p p p ppppppp p p p p p p p p p p p p p pp p p p p p p p p p p p p p pp pp p p p p p p p pp p p ppppPage 29425-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESGROUP10MG/M3ANIMAL#OBSERVATIONS1069WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED1070WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1-1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p ppppp p p p pp p p p p p p p p p p ppp p pPage 29525-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSMALESANIMAL#20512052205320542055205620572058GROUP22000MG/M3OBSERVATIONSWITHINNORMALLIMITSTERMINALSACRIFICEDAYOFSTUDYDERMALGENERAL:ALOPECIA-EXTREMITIES/SNOUTOCULAR:CHROMODACRYORRHEA-UNILATERALWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENT1-1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p p p pp3 3 3 3 3 3 3 3pp p p p p p p p p p p p p ppp p p p p p p p p p p p pppp p p p p p p p p p p p p ppp p p p p p ppp p p p p p pp p p p p p p p p p p p p pppp p p p ppp p p p p p p p p pp p p p p p p p p p p ppp p pPage 29625-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESANIMAL#20592060GROUP22000MG/M3OBSERVATIONSWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSREASSIGNEDTOSATELLITEGROUPORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 1 1 11 - 1 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p p p ppp p p p p p p p p p p p p pppPage 29725-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESANIMAL#305130523053305430553056305730583059GROUP310000MG/M3OBSERVATIONSWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEDERMALGENERAL:SCABSDERMALGENERAL:ALOPECIA-GENERALORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY/1 -1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p p ppp p p p p p p p p p p p pppp p p p p p p p p p p p p ppp p p p p p p p p p p p p ppp p p p p p p p p p p p p ppp p p p p p p p p p p p p pp pp p p p p p2 2 2pp p p pp p pp p p p p p p p p p p p p p pp p p p p p p p p p p p p ppppppPage 29825-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESANIMAL#3060GROUP310000MG/M3OBSERVATIONSWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 -1 1 1 112 2 3 4 4 5 6 6 7 8 9 9 901 1211 6 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p ppp pPage 29925-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESGROUP420000MG/M3ANIMAL#OBSERVATIONS4051WITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED4052WITHINNORMALLIMITSRECOVERYSACRIFICE4053WITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED4054WITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED4055WITHINNORMALLIMITSRECOVERYSACRIFICE4056WITHINNORMALLIMITSRECOVERYSACRIFICEOCULAR:CHROMODACRYORRHEA-BILATERALORAL/BUCCAL:NASALDISCHARGE-RED4057WITHINNORMALLIMITSRECOVERYSACRIFICE4058WITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 -1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p p p p p ppp pp p p p p p p p p p p p p p pp p p ppp p p p pp p p p p p p pp p p ppp pp p p p p p p p p p p p p p p p p pppp p p p p p p p p p p p p p p p p p ppp p p p p p pp p p p p p p ppp pp pp p p p p p p p p p p p p p p p p p ppp p p p p p p pp p p p p p ppp p p pPage 30025-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)MALESGROUP420000MG/M3APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSANIMAL#OBSERVATIONSDAYOFSTUDY1 1 1 11-1 2 2 34 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 24059WITHINNORMALLIMITSRECOVERYSACRIFICE4060WITHINNORMALLIMITSRECOVERYSACRIFICEDERMALGENERAL:ALOPECIA-EXTREMITIES/SNOUTORAL/BUCCAL:NASALDISCHARGE-CLEAR4061WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED4062WITHINNORMALLIMITSTERMINALSACRIFICEDERMALGENERAL:ALOPECIA-EXTREMITIES/SNOUTOCULAR:CHROMODACRYORRHEA-BILATERALORAL/BUCCAL:NASALDISCHARGE-REDORAL/BUCCAL:INCISORSMALOCCLUDED4063WITHINNORMALLIMITSTERMINALSACRIFICE4064WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED4065WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTp p p p p p p p p p p p p p pp p p ppp p p p p p p pp p pp3 3 3 3 3 3 3 3pp p p p p p p p p p p ppppp p pp3 3 3 3 3 3 3 3 3 3p p p p p ppp p p p p p p p pp p p p p p p p p p p p p ppp p pp ppp p p p p p pp pp p p p p p p p p p p p pppPage 30125-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSMALESANIMAL#4066GROUP420000MG/M3OBSERVATIONSWITHINNORMALLIMITSTERMINALSACRIFICE4067WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED4068WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED4069WITHINNORMALLIMITSTERMINALSACRIFICE4070WITHINNORMALLIMITSTERMINALSACRIFICEOCULAR:CHROMODACRYORRHEA-UNILATERALCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 1 1 11 - 1 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p p p ppp p p p p p p p p p p p ppp pp p p p p p p p p p ppp p p pp p p p p p p p p p p p p p ppp p p p p pp p p p p p p pppPage 30225-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)FEMALESGROUP10MG/M3APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 -1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 2ANIMAL#155115521553155415551556155715581559OBSERVATIONSWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEDERMALGENERAL:ALOPECIA-EXTREMITIES/SNOUTWITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICEWITHINNORMALLIMITSRECOVERYSACRIFICECODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENT1 1 6 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p p p p p p p ppp p p p p p p p p p pp p p ppp p p pp p p p p p p p p p p p p p p p p p ppp p p p p p p p p p pp2 2 2 2 2 2 2 2p p p p p p p p p p p p p p p p p pppp p p p p p p p p p p p p p p p p p ppp p p p p p p p p p p p p p p p p p ppp p p p p p p p p p p p p p p p p p ppp p p p p p p p p p p p p p p p p p ppPage 30325-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)FEMALESGROUP10MG/M3APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 -1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 2ANIMAL#1560OBSERVATIONSWITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED1561WITHINNORMALLIMITSTERMINALSACRIFICEOCULAR:CHROMODACRYORRHEA-UNILATERALORAL/BUCCAL:NASALDISCHARGE-RED1562WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED1563WITHINNORMALLIMITSTERMINALSACRIFICE1564WITHINNORMALLIMITSTERMINALSACRIFICE1565WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED1566WITHINNORMALLIMITSTERMINALSACRIFICEDERMALGENERAL:ALOPECIA-EXTREMITIES/SNOUTOCULAR:CHROMODACRYORRHEA-UNILATERAL1567WITHINNORMALLIMITSTERMINALSACRIFICEDERMALGENERAL:ALOPECIA-EXTREMITIES/SNOUTCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENT1 1 6 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p pp p p ppp pp p p p p p p p p ppp pp p pp p p p p p pp ppp p p p pp p p p p p p p p p p p p ppp p p p p p p p p p p p p ppp p p p ppp p p p p p p p pp p p p p p pp3 3 3 3 3 3 3 3p p pp p p p pp3 3 3 3 3 3 3 3 3 3Page 30425-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESGROUP10MG/M3ANIMAL#OBSERVATIONS1568WITHINNORMALLIMITSTERMINALSACRIFICE1569WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED1570WITHINNORMALLIMITSTERMINALSACRIFICECODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 1 1 11 - 1 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p p p ppp p p p p p p p p p p ppp p pp p p p p p p p p p p p p p ppPage 30525-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSFEMALESGROUP22000MG/M3ANIMAL#255125522553255425552556255725582559OBSERVATIONSWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEDERMALGENERAL:ALOPECIA-GENERALWITHINNORMALLIMITSTERMINALSACRIFICEDAYOFSTUDYORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEWITHINNORMALLIMITSTERMINALSACRIFICEOCULAR:CHROMODACRYORRHEA-UNILATERALORAL/BUCCAL:INCISORSMALOCCLUDEDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENT1 -1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p pp p p p p p p ppp pp p p p p p p p p p p p p ppp p p p p p p p p p p p p ppp p pp2 2 2 2 2 2 2 2 2 2 2p p p pp p p p p ppp p p pp p p p p p p p p p p p pp pp p p p p p p pp p p p p ppp p p p p p p p p p p p p p pp p p p p p p p p pp p p p pp p p p pppppPage 30625-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)FEMALESGROUP22000MG/M3ANIMAL#2560OBSERVATIONSWITHINNORMALLIMITSTERMINALSACRIFICEAPPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 1 1 11-1 2 2 34 4 5 6 6 7 8 9 9 901 1 21 16 307 4 15 2 9 6 301 7 4 18 2p p p p p p p p p p p p p p ppCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTPage 30725-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSFEMALESGROUP310000MG/M3ANIMAL#3551OBSERVATIONSWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED3552WITHINNORMALLIMITSTERMINALSACRIFICEDAYOFSTUDYDERMALGENERAL:ALOPECIA-EXTREMITIES/SNOUTORAL/BUCCAL:NASALDISCHARGE-RED3553WITHINNORMALLIMITSTERMINALSACRIFICE3554WITHINNORMALLIMITSTERMINALSACRIFICE3555355635573558WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENT1 -1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p p p ppp p pppp p pp p p p p pp3 3 3 3 3 3 3 3p pp p p p p p p p p p p p p ppp p p p p p p p p p p p p ppp p p p p p p p p p p p pppp p p p p p p p p p p ppp p pp p p p p p p p p p p p p pppp p p p p p p p p p p ppp p pPage 30825-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSFEMALESGROUP310000MG/M3ANIMAL#OBSERVATIONS3559WITHINNORMALLIMITSTERMINALSACRIFICEORAL/BUCCAL:NASALDISCHARGE-RED3560WITHINNORMALLIMITSTERMINALSACRIFICEDAYOFSTUDYDERMALGENERAL:ALOPECIA-EXTREMITIES/SNOUTORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENT1 1 1 11 - 1 2 2 3 4 4 5 6 6 7 8 9 9 901 1 21 16 307 4 1 85 2 9 6 301 7 4 18 2p p p p p p p p p p p p ppppp p pp2 2p p p p p p p p p pPage 30925-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)FEMALESGROUP420000MG/M3APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 -1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 901 1 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31025-SEP-200313:51HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)APPENDIXBGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSFEMALESGROUP420000MG/M3ANIMAL#4558455945604561OBSERVATIONSWITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-REDWITHINNORMALLIMITSRECOVERYSACRIFICEORAL/BUCCAL:NASALDISCHARGE-CLEARWITHINNORMALLIMITSTERMINALSACRIFICE4562WITHINNORMALLIMITSTERMINALSACRIFICE4563WITHINNORMALLIMITSTERMINALSACRIFICEOCULAR:CHROMODACRYORRHEA-BILATERALOCULAR:CHROMODACRYORRHEA-UNILATERALORAL/BUCCAL:NASALDISCHARGE-REDORAL/BUCCAL:INCISORSMALOCCLUDED4564WITHINNORMALLIMITSTERMINALSACRIFICE4565WITHINNORMALLIMITSTERMINALSACRIFICEOCULAR:LACRIMATION-UNILATERALORAL/BUCCAL:NASALDISCHARGE-REDCODE:1-SLIGHT2-MODERATE3-MARKEDP-PRESENTINDIVIDUALCLINICALOBSERVATIONSDAYOFSTUDY1 -1 1 1 11 2 2 3 4 4 5 6 6 7 8 9 9 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33325-SEP-200313:55HuntingdonLifeSciences00-6130ToxicitySub-Group(MainandRecovery)MALESGROUP10MG/M3ANIMAL#PARTOFEYE106510661067106810691070APPENDIXCGASOLINEDIPEVAPORCONDENSATE:A13-WEEKWHOLE-BODYINHALATIONTOXICITYSTUDYINRATSWITHNEUROTOXICITYASSESSMENTSAND4-WEEKINVIVOGENOTOXICITYANDIMMUNOTOXICITYASSESSMENTSINDIVIDUALOPHTHALMOLOGYOBSERVATIONSOBSERVATIONNOVISIBLELESIONSNOVISIBLELESIONSNOVISIBLELESIONSNOVISIBLELESIONSNOVISIBLELESIONSNOVISIBLELESIONSWEEK12Page 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