+ A e> P. O. Box 331, Millwood, VA 22646 (540) 837 1602 Task Fort June 7,2002 Document Control Office (7407) Office of Pollution Prevention and Toxics Room G 099 Environmental Protection Agency 401 M Street, S. W. Washington, D. C. 20460 Re: Testing Consent drder for 1,1,2 Trichloroethane; Final Reports of Subchronic Inhalation Toxicity and Pharmacokinetic Model Development Studies (Docket No. OPPTS 42 198Cj Dear Sirs:

Pursuant to Table 1 of the Enforceable Consent Agreement for 1,1,2 Trichloroethane, the HAP Task Force submits the enclosed final reports: 4  "A 90 Day Inhalation Study of 1,1,2 Trichloroethane (I, 1,2 TCE) in Rats. (With Satellite Groups for Pharmacokinetic Evaluations in Rats and Mice)" WIL Research Laboratories, Inc.

"Physiologically Based Pharmacokinetic Model Development, Simulations, and Sensitivity Analysis for Repeated Exposure to 1,1,2 Trichloroethane" by The Sapphire Group Six copies of each report are enclosed. Please do not hesitate to let me kno& there are any questions. G, CD 9 % zg ~~~~~~~~~~~~ 11~~~~~~  <;~ y& y/,~ ; $rr Peter E. Voytek, Pi. D. 4 ;; Manager +

Enclosures C. Auer (w/ out enclosures) R. Leukroth (w/ out enclos& es) J. Schaeffer W. C. Norman, Esq. FINAL REPORT Volume 1 of 2 (Text, Summary and Individual Tables l 3 1)

STUDY TITLE A 90 Day Inhalation Toxicity Study of 1, 1,2 Trichloroethane (1, 1,2 TCE) in Rats (With Satellite Groups for Pharmacokinetic Evaluations in Rats and Mice)

STUDY NUMBER WIL 4 17002 DATA REQUIREMENT U. S. EPA TSCA 799.9346 STUDY DIRECTOR Daniel T. Kirkpatrick, Ph. D., D. A. B. T.

STUDY INITIATION DATE May 15,200l STUDY COMPLETION DATE May 23,2002 PERFORMING LABORATORY WIL Research Laboratories, Inc. 1407 George Road Ashland, OH 44805 928 1 SPONSOR HAP Task Force P. O. Box 331 Millwood, VA 22646 1 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane COMPLIANCESTATEMENT This study, designated WIL 417002, was conducted in compliance with the Environmental Protection Agency Toxic Substances Control Act (EPA TSCA) Good Laboratory Practice ReguIations (40 CFR Part 792), the standard operating procedures of WIL Research Laboratories, Inc., and the protocol as approved by the sponsor. The protocol was designed to generally conform with the EPA TSCA Guideline 799.9346:

90 Day Inhalation Toxicity Study. Analyses of test article samples were performed by the sponsor (presented in Appendix A); it is unknown whether the characterization analyses were conducted according to Good Laboratory Practices (GLP).

Study Difector 2 of 619 . WIL 4 17002 1, 1,2 Trichloroethane HAP Task Force TABLE OF CONTENTS VOLUME 1 w Compliance Statement .............................................................................. 2 Table Of Contents ..................................................................................... 3 Index Of Tables ......................................................................................... 6 Index Of Appendices ................................................................................. 8 Summary .................................................................................................... 9 Objective ..................................................................................................... 9 Study Design ............................................................................................... 9 Results ....................................................................................................... 10 Conclusions ............................................................................................... 11 Introduction ............................................................................................. 12 General Study Information ........................................................................ 12 Key Study Dates ........................................................................................ 12 Study Design ............................................................................................ 13 Experimental Procedures  Materials And Methods ........................... 14 Test Article ................................................................................................ 14 Test Article Identification ......................................................................... 14 Exposure Methods.. ................................................................................... 14 Generation Of Exposure Atmosphere ....................................................... 16 Characterization Of Exposure Atmospheres ............................................. 16 Organization And Treatment Regimen ..................................................... 16 Animal Receipt And Acclimation/ Pretest Period.. .................................... 19 Animal Housing ........................................................................................ 19 Diet, Drinking Water And Maintenance ................................................... 20 Environmental Conditions ........................................................................ 20 Assignment Of Animals To Treatment Groups ....................................... .20 Data Retention ........................................................................................... 21 1.

1.1.

1.2.

1.3.

1.4.

2.

2.1.

2.2.

3.

4.

4.1.

4.1.1.

4.1.2.

4.1.3.

4.1.4.

4.1.5.

4.2.

4.3.

4.4.

4.5.

4.6.

4.7.

3 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane VOLUME 1 (continued) Page 5.

5.1.

5.2.

5.3.

5.4.

5.4.1.

5.4.2.

5.5.

5.6.

5.7.

5.7.1.

5.7.2.

5.7.3.

5.8.

6.

6.1.

6.2.

6.3.

6.4.

6.4.1.

6.4.2.

6.5.

6.6.

6.6.1.

6.6.2.

6.6.3.

7.

8.

9. Parameters Evaluated ............................................................................. 22 Clinical Observations And Survival (All Groups) ................................... 22 Body Weights (All Groups) ..................................................................... .22 Food Consumption (Toxicology Groups) ................................................ .22 Clinical Pathology (Toxicology Groups) .................................................. 23 Hematology ............................................................................................... 23 Serum Chemistry ....................................................................................... 24 Plasma Level Determinations (Pharmocokinetic Groups) ........................ 24 Ophthalmic Examinations (Toxicology Groups) ..................................... .25 Anatomic Pathology (Toxicology Groups). .............................................. 25 Macroscopic Examination ......................................................................... 25 Organ Weights ........................................................................................... 27 Slide Preparation And Microscopic Examination.. .................................. .27 Statistical Methods .................................................................................... 27 Results And Discussion (Toxicology Groups) ....................................... 29 Clinical Observations And Survival .......................................................... 29 Body Weights ............................................................................................ 29 Food Consumption .................................................................................... 30 Clinical Pathology ..................................................................................... 30 Hematology ............................................................................................... 30 Serum Chemistry.. ..................................................................................... 3 1 Ophthalmic Examinations.. ....................................................................... 3 1 Anatomic Pathology .................................................................................. 31 Macroscopic Examination.. ....................................................................... 3 1 Organ Weights.. ......................................................................................... 32 Microscopic Examination ......................................................................... 32 Conclusions .............................................................................................. 37 Key Study Personnel And Report Submission ..................................... 40 Quality Assurance Unit Statement ........................................................ 41 4of619 WIL 417002 HAP Task Force 1, 1,2 Trichloroethane VOLUME 1 (continued) &

9.1. Phases Inspected.. ...................................................................................... 41 9.2. Approval .................................................................................................... 43 10. References ................................................................................................ 44 11. Deviations From The Protocol ............................................................... 46 5 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane INDEX OF TABLES VOLUME 1 (continued) Page 1~ Summary Of Survival And Disposition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48 2. Summary Of Clinical Findings: Total Occurrence/ No. Of Animals (Detailed Physical Examinations/ Dispositions) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 3. Summary Of Clinical Findings: Total Occurrence/ No. Of Animals (Prior To Exposure) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 4. Summary Of Clinical Findings: Total Occurrence/ No. Of Animals (Following Exposure) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 5. Summary Of Clinical Findings: Total Occurrence/ No. Of Animals (Non Exposure Days) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5 8 6. Summary Of Body Weights [G] ................................................................................ .60 7. Summary Of Body Weight Changes [G] .................................................................. .68 7A. Summary Of Cumulative Body Weight Changes [G] .............................................. .74 8. Summary Of Weekly Food Consumption [G/ Animal/ Day] ...................................... 80 9. Summary Of Hematology Values .............................................................................. 86 10. Summary Of Leukocyte Differential Counts [%] ...................................................... 90 11. Summary Of Leukocyte Counts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92 12. Summary Of Serum Chemistry Values . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 13. Summary Of Ophthalmic Examination Findings (Week  1 Pretest Evaluation) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 14. Summary Of Ophthalmic Examination Findings (Week 12 Evaluation) . . . . . . . . . . . . . . . . 103 15. Gross Necropsy Observations Incidence Summary (Unscheduled Death) . . . . . . . . . . ... 104 16. Gross Necropsy Observations Incidence Summary (Primary Necropsy). .............. .105 17. Organ Weights (Grams)  Summary Of Means ...................................................... .107 6of619 WIL 417002 HAP Task Force 1, 1,2 Trichloroethane VOLUME 1 (continued) &

18. Organ Weights Relative To Final Body Weights (Grams/ 100 Grams) . . . . . . . . . . . . . . . . . . . 112 19. Organ Weights Relative To Brain Weights (Grams/ 100 Grams) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118 20. Individual Survival And Disposition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 2 1. Individual Body Weights [G] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127 22. Individual Body Weight Changes [G] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143 23. Individual Food Consumption [G/ AnimaVDay] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159 24. Individual Hematology Values.. .............................................................................. .175 25. Individual Leukocyte Differential Counts [" A]. ....................................................... .183 26. Individual Leukocyte Counts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 1 27. Individual Serum Chemistry Values.. ...................................................................... .199 28. Individual Ophthalmic Examination Findings (Week 1 Pretest Evaluation). ...... .. 215 29. Individual Ophthalmic Examination Findings (Week 12 Evaluation) ................... .2 19 30. Individual Macroscopic Findings (Unscheduled Death). ........................................ .223 3 1. Individual Macroscopic Findings (Primary Necropsy) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 224 VOLUME 2 32. Individual Organ Weights And Final Body Weights [G] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305 33. Individual Organ Wts. Relative To Final Body Wts. [G/ 100 G] ............................ .3 17 34. Individual Organ Weights Relative To Brain Weights [G/ 100 G] ......................... .329 7 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane INDEX OF APPENDICES A.

B.

C.

D.

E.

F.

G.

H.

I.

J.

K.

L. VOLUME 2 (continued) Paae Analysis of Test Article Samples (Sponsor Provided) . . . . . . . . . . . . . . . . . . . . . . . . . . . .. I.............. 341 Purity Assessment (WIL Research Laboratories, Inc.) ..**...................................,.... 353 Test Atmosphere Generation and Environmental Conditions During Exposure (WIL Research Laboratories, Inc.) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366 Clinical Pathology Parameters For Control Animals (Study Week 8) . . . . . . . . . . . . . . . . . . . . 414 Pretest Clinical Observations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4 19 Serological Screening (BioReliance Corporation) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 423 Clinical Pathology Methods, Procedures and References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 427 Individual Clinical Observations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . , , . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .432 Dispositions and Body Weights (Pharmacokinetic Groups) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 493 Ophthalmic Examination Report (David A. Wilkie, D. V. M., M. S., D. A. C. V. O.) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 502 Histopathology Report (Experimental Pathology Laboratories, Inc.) . . . . . . . . . . . . . . . . . . . . . 504 Study Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 585 8 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 1. SUMMARY 1.1. OBJECTIVE The objective of the study was to evaluate potential subchronic toxic effects of 1, 1, 2 Trichloroethane when administered via whole body inhalation to rats for 90 days.

In addition, 36 female F 344 rats and 36 female B6C3Fl mice were exposed by whole

body inhalation for 28 days and used for a pharmacokinetic study.

1.2. STUDY DESIGN Potential subchronic toxic effects of the test article, 1, 1, 2 Trichloroethane, were evaluated in this 90 day study in rats. The test article was administered via whole body inhalation as a vapor to three groups (Groups 2 4), each comprised of 10 male and 10 female Fischer 344 CDF@( F 344) Crl: BR rats, five days per week, as a six hour exposure,

for 13 consecutive weeks (minimum of 65 exposures). The targeted exposure concentrations were 15, 40 and 100 ppm. The exposure concentrations were measured by chromatographic methods and were found to be 15, 40 and 100 ppm. A concurrent control group' (Group 1) of identical design was exposed to clean, filtered air on a comparable regimen. Clinical examinations were performed daily and detailed physical examinations were conducted weekly. Individual body weights and food consumption were recorded weekly. Ophthalmic examinations were conducted prior to exposure initiation and near exposure termination. Clinical pathology evaluations (hematology and serum chemistry) were conducted at the end of the exposure period. Complete necropsies were performed on all animals and selected organs were weighed. Selected tissues were examined microscopically.

For the pharmacokinetic assessment, 36 female Fischer 344 CDF@( F 344) Crl: BR rats and 36 female B6C3FlKrl: BR mice (Groups 5 and 6, respectively), were exposed five days per week, as a six hour exposure, for four consecutive weeks (minimum of 20 exposures). The targeted exposure concentration was 100 ppm. Blood samples were collected at 4, 6, 6.25 and 8 hours after the start of exposure from three animals/ specie/ timepoint on the first, third and fifth days of study week 4. Following 9 of 619 WIL 417002 HAP Task Force 1, 1,2 Trichloroethane blood collection, the anesthetized animals were exsanguinated and organs/ tissues were collected and were frozen using liquid nitrogen for potential analysis.

1.3. RESULTS There were no test article related deaths. Clinical observations, body weights, food consumption, hematology parameters and organ weights were unaffected by test article exposure. No test article related ophthalmic findings were noted, and no test article

related macroscopic changes were observed at the scheduled necropsy.

Test article related effects noted in the 15 ppm group consisted of:

l Minimal hepatocellular vacuolization in 3/ l 0 females.

l Microscopic lesions in the olfactory epithelium of the nasal turbinates (vacuolization'microcyst formation in 2/ l 0 males and 4/ l 0 females).

Test article related effects noted in the 40 ppm group consisted of:

l Minimal hepatocellular vacuolization in 3/ 10 females.

l Microscopic lesions in the olfactory epithelium of the nasal turbinates (atrophy in 6/ 10 males and 7/ 10 females, vacuolizatiommicrocyst formation in 6/ 10 males and 4/ 10 females and respiratory epithelial metaplasia in l/ 10 males and l/ 10 females).

Possible test article related effects noted in the 100 ppm group consisted of:

l Decreased absolute body weights and body weight gains from study week 0 to 1 in females.

l Increased mean cholesterol levels in the males and females and decreased mean glucose in females.

l Minimal hepatocellular vacuolization in 5/ l 0 males and 1 O/ l 0 females.

l Microscopic lesions in the olfactory epithelium of the nasal turbinates (atrophy in 7/ l 0 males and lo/ 10 females, vacuolization'microcyst formation in lO/ lO males and 8/ 10 females and respiratory epithelial metaplasia in 300 males and 5/ 10 females).

lOof WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 1.4. CONCLUSIONS Based on the results of this study, the no observed effect level (NOEL) for whole body inhalation of 1, 1, 2 Trichloroethane to rats for 13 weeks was less than 15 ppm.

However, the no observed adverse effect level (NOAEL) was less than 40 ppm since the hepatic and nasal effects observed for the low concentration group were not considered toxicologically significant. In addition, a major difference in chemical sensitivity of the nasal airways between the rat and human may exist because of the vastly greater presence of olfactory epithelium in the rat.

11 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 2. INTRODUCTION 2.1. GENERAL STUDY INFORMATION This report presents the data from "A 90 Day Inhalation Toxicity Study of 1, 1,2 Trichloroethane (1, 1, 2 TCE) in Rats (With Satellite Groups for Pharmacokinetic Evaluations in Rats and Mice)". Due to software spacing constraints, the study title appears as, "90 Day Inhal. Study of 1, 1, 2 TCE in Rats with PK in Rats and Mice" on the report tables.

The following computer protocols were used for data collection during the study:

Computer Protocol Type of Data Collected WIL 4 17002.. .......................................... Main study data WIL 4 17002R.. ....................................... Rat pharmacokinetic group data WIL 417002M.. ...................................... Mouse pharmacokinetic group data WIL 4 17002P ......................................... Pretest rat data WIL 4 17002Q.. ....................................... Pretest mouse data 2.2. KEY STUDY DATES Date( s) Event( s)

May 15,200l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Experimental starting date (animal receipt) May 29,200l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Assignment to study groups (Toxicology Phase) June 1,200l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Assignment to study groups (Pharmacokinetic Phase) May 31, 2001 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Initiation of dosing for Toxicology Phase (study week 0) June 5,200l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Initiation of dosing for Pharrnacokinetic Phase (study week 0) June 25, 27 and 29, 2001 . . . . . . . . . . . . . . . . . . . . . . . . Pharmacokinetic blood collection August 30 and 3 1, 200 1 . . . . . . . . . . . . . . . . . . . . . . . . . . Primary necropsy (study week 13) December 3 1, 2001 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Experimental termination date (last histopathological examination)

12of619 . WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 3. STUDY DESIGN Acclimation and Pretest Period Assignment of Animals to Study Groups 10 Animals/ Sex/ Group (1  4) Treated Six Hours/ Day, 5 Days/ Week for 13 Weeks for a Minimum of 65 Exposures I I Clinical Observations Recorded / Daik;; zeFod Clinical Pathology Evaluations Conducted at Appropriate Intervals pzz? QkJ 36 Females/ Specie/ Group (5  6)

Clinical Observations Recorded Daily; Body Weights Recorded as Appropriate / Blood Samples for Plasma Level 1 DeteArm$ atiF gll;;; d at Ophthalmic Examinations Performed at Appropriate Intervals I I Animals Found Dead or Euthanized 1 Necropsies Performed on All Surviving Animals/ Sex/ Group During Week 13; Selected Organs Weighed; Selected Tissues Examined Microscopically 13 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 4. EXPERIMENTALPROCEDURES MATERIALS AND METHODS 4.1. TESTARTICLE 4.1.1. TESTARTICLEIDENTIFICATION The test article, 1, 1, 2 Trichloroethane, was received from Dow Chemical, Freeport,

Texas, as follows:

Identification Quantity Physical Received Description Date of Receint 1, 1,2 Trichloroethane Lot No. NH06019301 [WIL Log No.: 5015A] 3 Buckets Total gross weight: 81059. Og Clear, colorless liquid 12/ 29/ 01 The purity of the test article was 99.55% by weight based on the analyses of the test article samples provided by WIL Research Laboratories, Inc. to the sponsor (Appendix A). Purity (Appendix B) and stability data for the test article were conducted by the Analytical Chemistry Department, WIL Research Laboratories, Inc. The test article was stored at room temperature, and was considered stable under these conditions. A reserve sample of the test article (11.7 grams) was collected on February 20, 200 1, and stored in the Archives of WIL Research Laboratories, Inc.

4.1.2. EXPOSURE METHODS The exposures were conducted in 2 m3 stainless steel and glass whole body exposure chambers. One chamber was dedicated for each group for the duration of the study. For the high concentration groups, the toxicology and pharrnacokinetic group rats (with the exception of blood collection days, discussed below) and pharmacokinetic group mice were exposed in the same inhalation chamber. This is considered to be acceptable for 6 hour exposures and for the short duration of the pharmacokinetic study (28 days). The chambers were operated under dynamic conditions from a HEPA and charcoal filtered air source.

Animal cage batteries were rotated on a daily basis through various chamber locations in order to reduce the potential differences in exposure due to any residual non homogeneity in chamber concentrations.

14 of 619 WIL 417002 HAP Task Force 1, 1,2 Trichloroethane Air flow rate through the chamber was such that there was at least 10 air changes per hour.

Protocol defined ranges for daily average chamber temperature and relative humidity were 22k2" C and 30 70%, respectively. These parameters were monitored continuously and recorded approximately every 30 or 35 minutes during each day's exposure. Oxygen content of the exposure atmosphere was measured during the methods developmental phase of the study and was at least 19%.

The animals were caged individually, without food and water, during the exposures with the exception of the mice in the pharmacokinetic phase, which were allowed access to water. The exposure period was six hours per day, five days per week for 13 consecutive weeks (minimum of 65 exposures). For each exposure, all animals were housed in normal animal colony rooms during non exposure hours. For each day's exposure, the animals were transferred to exposure caging, transported to the exposure chamber room, exposed for the requisite duration and returned to their home cages. The control group was exposed to clean, filtered air under conditions otherwise identical to the test article treated groups.

Mean exposure temperature ranged from 21 to 25° C and relative humidity ranged from 42 to 49%. Exposure methods and conditions are detailed in Appendix C.

On each day of PK blood collection (lS*, 3rd and 5' days of exposure of the fourth exposure week), the female rats designated for blood collection on the specific day were exposed to the target concentration of 100 ppm in a 500 L NYU type whole body chamber (that is,

separate from the remaining animals in the group). This was a completely separate exposure system with its own test atmosphere vapor generator, chamber dilution airflow and calibrated exhaust airflow. The methods used for this generation and exposure system were similar to those for the other test article systems. This change for the pharmacokinetic (PK)

study rats facilitated blood collection at the appropriate time points relative to the start of exposure and prompt collection of blood after animals were removed from the exposure chamber (4 and 6 hour time points). The use of different exposure systems for PK study rats and mice allowed the PK rat exposure to be started approximately 30 minutes after or 15of619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane before the mouse exposure. Exposure methods and conditions are detailed in Appendix C.

4.1.3. GENERATION OF EXPOSURE ATMOSPHERE Vapors of the test article were generated using a glass bead vaporization system. The vaporization column consisted of a glass tube filled with glass beads. Compressed air was metered to the bottom of the column using calibrated flowmeters. Vaporization occurred as the test article flowed down through the beads and the compressed air flowed up through the column. The concentrated vapors were piped to the chamber inlet where the concentration was reduced to the desired level by mixing with the chamber ventilation air. The vaporization columns were heated by heating tapes operated from Omega CN370 temperature controllers. Exposure methods and conditions are detailed in Appendix C.

4.1.4. CHARACTERIZATION OF EXPOSURE ATMOSPHERES Nominal exposure concentrations were determined by weighing the containers of test article prior to and again after each day's exposure, and calculating the concentration from the weight difference and the total volume of air passed through each chamber during that day's exposure.

Actual exposure concentrations were determined approximately every 30 or 35 minutes using a gas chromatographic method. Exposure methods and conditions are detailed in Appendix C.

4.1.5. ORGANIZATION AND TREATMENT REGIMEN The study consisted of six groups. The toxicological phase of the study consisted of four groups of Fischer 344 CDF@( F 344) Crl: BR rats which were exposed five days per week,

as a six hour exposure, for 13 consecutive weeks (minimum of 65 exposures). The pharmacokinetic phase consisted of one group of female Fischer 344 CDF@( F 344) Crl: BR rats and one group of female B6C3FlKrl: BR mice which were exposed five days per week, as a six hour exposure, for four consecutive weeks .

16 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane The following table presents the study group assignment:

Toxicology Groups (WIL 4 17002)

Group Number Test Article Svecie Target Exposure Number of Animals Level (pvm) Males Females 1 Filtered Air Rat 0 10 10 2 Low 1 ,l ,2 TCE Rat 15 10 10 3 Mid 1,1,2 TCE Rat 40 10 10 4 High 1,1,2 TCE Rat 100 10 10 Toxicokinetic Groups (WIL 4 17002R [Rat1 4 17002M IMousel)

Group Target Exposure Number of Animals Number Test Article Specie Levela (ppm) Males Females 5 High 1,1,2 TCE Rat 100 36 6 High 1,1,2 TCE Mouse 100 36 a = Groups 5 and 6 were exposed using the same exposure system and methods used for Group 4 (except for Group 5 PK animals on day of animals' blood collection).

Exposure concentrations were selected based on results from a prior dose range finding study (4 1700 1').

The selected route of administration was whole body inhalation exposure as this is a potential route of human exposure. The animal models, the Fischer 344 CDF@( F

344) Crl: BR rat and the B6C3Fl/ Crl: BR mouse, are recognized as appropriate for subchronic inhalation toxicity and pharmacokinetic studies and are recommended in the TSCA guideline.

On July 26, 2001 (study day 56), Groups 1 and 2 were loaded into the incorrect cage batteries. This error resulted in the control group animals (Group 1) receiving a single six hour exposure to 15 ppm 1, 1, 2 TCE and the Group 2 (15 ppm group) animals receiving a six hour exposure to filtered air. The error was discovered upon returning the animals to their home cages. Although this error would have been avoided if the single responsible individual followed normal verification procedures, a second, independent 17 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane verification step was added and all remaining exposures were completed without further exposure errors.

At the time of this incident, there was approximately five weeks of scheduled exposures remaining prior to conduct of the primary toxicity evaluations. Since the study was designed to evaluate potential subchronic effects of 1,1,2 TCE following repeated six

hour, five day/ week exposures for 13 weeks, it is considered unlikely that a single exposure of the control animals to the low level of 15 ppm would affect the study outcome. Even if a single exposure to 15 ppm had some slight acute effect, complete resolution would be expected during the approximately five weeks remaining before end of the exposure phase. Previous toxicity testing of 1,1,2 TCE cited in Patty's Industrial Hygiene and Toxicology indicated that inhalation exposure of rats to 15 ppm for six months (7 hours/ day, 5 days/ week) had no toxic effects as indicated by all standard endpoints2.

In order to ensure that there was no significant acute effect on the control group animals,

blood was collected from six males and six females in the control group at approximately 18 hours following the exposure (presented in Appendix D). In addition, blood was collected from five untreated male Fischer 344 rats from the original shipment (no untreated females from the original shipment were available). Serum enzyme levels,

including alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase levels were determined to evaluate possible effects on the liver since this was a primary target tissue of 1, 1, 2 TCE toxicity. The results of the serum enzyme evaluation did not suggest an effect that would interfere with the interpretation of the results of this study As will be described in the results and conclusions sections of this report (sections 6 and 7), the results of this study support the conclusion that the single exposure of the control group at 15 ppm did not affect the study interpretation or objectives. Since there were no adverse effects for the animals in the 15 ppm group following 13 weeks of exposure, no 18 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane adverse or confounding effects would be present approximately five weeks following a single exposure.

4.2. ANIMAL RECEIPT Am ACCLIMATION/ PRETEST PERIOD Fifty male and eighty six female Fischer 344 CDF@( F 344) Crl: BR rats and forty seven female B6C3Fl mice in good health were received on May 15, 2001, from Charles River Laboratories, Raleigh, North Carolina and Portage, Michigan, respectively. The animals were 41 days old at receipt. Each animal was examined by a qualified technician on the day of receipt and weighed three days later. Animals were uniquely identified by Monel@

metal ear tags (rats) or by a tail tattoo (mice) displaying the permanent identification number. All animals were housed for a minimum 16 day acclimation period. During this period, each animal was observed twice daily for mortality and general changes in appearance or behavior.

Serological screening was conducted on five rats of each sex and five female mice approximately one week following receipt. Blood was collected via the vena cava from the anesthetized animals. The animals were then exsanguinated and discarded. Serum samples for rats were analyzed for antibodies to the following pathogens: Sendai,

RCVSDA, Reo, KRV, LCM, H l, PVM, Ropv viruses, M pulmonis and CARB. Serum samples for mice were analyzed for Sendai, EDIM, GDVII, MHV, PVM, MPV, MVM and M pulmonis. The results are presented in Appendix F.

Pretest data collection began on May 18, 2001. Individual body weights were recorded and detailed physical examinations were performed four times for the rats and four times for the mice prior to the initiation of exposure. Food consumption, clinical pathology parameters and ophthalmic examinations were also recorded for pretest animals prior to the initiation of dosing. Pretest clinical observations are presented in Appendix E.

4.3. ANIMAL HOUSING Upon arrival, all animals were housed individually in clean, wire mesh cages suspended above cage board. Animals were maintained in accordance with the "Guide for the Care 19 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane and Use of Laboratory Animalstt3. The animal facilities at WIL Research Laboratories,

Inc., are accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC International).

4.4. DIET, DRINKING WATER AND MAINTENANCE The basal diet used in this study, PM1 Nutrition International, Inc. Certified Rodent LabDiet@ 5002, is a certified feed with appropriate analyses performed by the manufacturer and provided to WIL Research Laboratories, Inc. Municipal water supplying the facility is sampled for contaminants according to the standard operating procedures. The results of the diet and water analyses are maintained at WIL Research Laboratories, Inc. No contaminants were present in animal feed or water at concentrations sufficient to interfere with the objectives of this study. Reverse osmosis

treated (on site) drinking water, delivered by an automatic watering system, and the basal diet were provided ad Zibitum throughout the study (except during exposures for rats and during exposures for mice which had access to water only).

4.5. ENVIRONMENTAL CONDITIONS All animals were housed throughout the acclimation period and during the study in an environmentally controlled room. Controls were set to maintain a temperature of 72 f 4° F (22 & 3° C) and a relative humidity of approximately 30 70%. Room temperature and relative humidity were monitored continuously and recorded daily. The actual temperature ranged from 70.1" F to 74.2" F (21.2" C to 23.4" C) and humidity ranged from 39.6% to 62.2% during the study. Light timers were set to provide a 12 hour light/ l2 hour dark photoperiod. The 12 hour light/ l2 hour dark photoperiod was interrupted as necessary to allow for the performance of protocol specified activities.

4.6. ASSIGNMENT OF ANIMALS To TREATMENT GROUPS Two and four days prior to exposure (May 29, and June 1,2001, for the main study and the pharmacokinetic phase, respectively), all available animals were weighed using the WIL Toxicology Data Management System (WTDMSTM) and examined in detail for 20of619 WIL 417002 HAP Task Force 1, 1,2 Trichloroethane physical abnormalities. These data were reviewed by the study director and animals judged suitable for assignment to the study were selected for use in the computerized randomization procedure. The main study was randomized by stratification in a block design. The randomization for the pharmacokinetic groups was conducted using an unstratified method for the mice and the remaining rats. A printout containing the animal numbers, corresponding body weights and individual group assignments was generated.

The animals then were arranged according to the printout. For the toxicology groups, the control and 15, 40 and 100 ppm groups (Groups 1  4, respectively) each consisted of 10 males and 10 females. Each pharmacokinetic group (Groups 5 and 6) consisted of 36 females. The selected animals were approximately eight weeks old at the initiation of dosing; body weight values ranged from 153 g to 191 g for males and from 118 g to 140 g for females in the toxicology groups and from 122 g to 143 g for rats and from 19.3 g to 21.9 g for mice in the pharmacokinetic groups. Individual body weights at randomization were within + 20% of the mean for each sex.

4.7. DATA RETENTION The sponsor has title to all documentation records, raw data, specimens or other work product generated during the performance of the study. All work product including raw paper data and specimens are retained in the Archives at WIL Research Laboratories,

Inc., as specified in the study protocol.

The raw data, reserve samples of the test and control articles, if appropriate, and the original final report are retained in the Archives at WIL Research Laboratories, Inc., in compliance with regulatory requirements.

21 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 5. PARAMETERSEVALUATED 5.1. CLINICALOBSERVATIONSANDSURVIVAL( ALLGROUPS)

All animals were observed twice daily, once in the morning and once in the afternoon, for mortality and moribundity.

For the toxicology group animals, clinical examinations were performed twice daily on exposure days, prior to exposure and O 2 hours following exposure (designated as 2 hours post dosing for report presentation purposes). On non exposure days, clinical observations were performed once in the morning. For the pharmacokinetic animals,

clinical observations were performed prior to exposure and on non exposure days they were performed once in the morning. All significant findings were recorded. Detailed physical examinations were conducted on all main study animals weekly, beginning one week prior to randomization, on the day of randomization, weekly during the exposure period and prior to the scheduled necropsy. In addition, a detailed physical examination was conducted on all animals that were euthanized in extremis, to assist in the determination of the cause of death.

5.2. BODYWEIGHTS( ALLGROWS)

Individual body weights were recorded weekly, beginning at least one week prior to test article administration (study week 1). Mean body weights and mean body weight changes were calculated for the corresponding intervals. Final body weights (fasted for rats) were recorded prior to each scheduled necropsy. Individual body weights for the toxicokinetic group animals are presented in Appendix G.

5.3. FOODCONSUMPTION( TOXICOLOGYGROUPQ Individual food consumption was recorded weekly, beginning at least one week prior to test article administration (study week 1). Food intake was calculated as g/ animal/ day for the corresponding body weight intervals.

22 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 5.4. CLINICALPATHOLOGY( TOXICOLOGYGROWS)

Blood samples for clinical pathology evaluations (hematology and serum chemistry) were collected from all animals at the scheduled necropsy (study week 13). The animals were fasted overnight prior to the collection of blood samples. Blood was collected from the vena cava at the time of necropsy from animals under deep anesthesia from isoflurane inhalation, Clinical pathology methods, procedures and references are presented in Appendix G. In addition, blood samples were collected from the control animals during study week 8 and analyzed for possible effects on the liver (Appendix D).

The following parameters were evaluated.

5.4.1. HEMATOLOGY Total Leukocyte Count (White Cell) Prothrombin Time (Pro Time) Erythrocyte Count (Red Cells) Activated Partial Thromboplastin Time Hemoglobin (APTT) Hematocrit Differential Leukocyte Count  Mean Corpuscular Volume (MCV) Percent and Absolute Mean Corpuscular Hemoglobin Neutrophil (MCH) Lymphocyte Mean Corpuscular Hemoglobin Monocyte Concentration (MCHC) Eosinophil Platelet Count (Platelet) Basophil ( )  Designates tabular abbreviation 23 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 5.4.2. SERUM CHEMISTRY Albumin Aspartate Aminotransferase Total Protein (Aspartat Transfer) Globulin [by calculation] Gamma Glutamyltransferase" Albumin/ Globulin Ratio (Glutamyl Transfer) (A/ G Ratio) [by calculation] Glucose Total Bilirubin (Total Bili) Total Cholesterol (Cholesterol) Urea Nitrogen Calcium Creatinine Chloride Alkaline Phosphatase Phosphorus (Alkaline Phos'tse) Potassium Alanine Aminotransferase Sodium (Alanine Transfer) Triglycerides (Triglyceride)

( )  Designates tabular abbreviation a  Formerly gamma glutamyl transpeptidase 5.5. PLASMA LEVEL DETERMINATIONS (PJXARMOCOKINETIC GROIJPS)

Blood samples for test article plasma level determination were collected on the first, third and fifth days of the fourth week at 4, 6, 6.25 and 8 hours following the exposure start.

Three animals/ specie were sampled at each time point. To accommodate the 4 hour timepoint, the high concentration chambers were opened quickly, the designated animals (three rats and three mice) were removed and the chambers were resealed. The animals were anesthetized as quickly as possible and blood was collected via the vena cava into hepranized syringes. The blood was transferred in duplicate directly into headspace vials for analysis, and the remaining sample was collected and saved in vacutainers with dry sodium heparin. For the headspace vial samples, duplicate blood samples of 100 l.. tL from each animal were aliquoted into cold, pre weighed 20 mL headspace vials with crimp tops and PTFE silicone septa. The vials were sealed as quickly as possible. The headspace vials were weighed before and after the addition of blood to verify volumes added gravimetrically. Immediately after weighing, the vials were placed on dry ice. For each animal, the remaining blood was placed in a pre weighed, pre cooled vacutainer tube, which was sealed and reweighed.

24 of 619 WIL 417002 HAP Task Force 1, 1,2 Trichloroethane Following blood collection, the anesthetized animals were exsanguinated to complete euthanasia. The lungs, liver, kidneys, spleen and whole brain were collected at stored at 70° C at WIL Research Laboratories, Inc. In order to minimize chemical loss, all sample manipulations were conducted on ice. Each organ was double wrapped in plastic wrap,

placed in a labeled aluminum, foil pouch, frozen in liquid nitrogen and sealed in a plastic storage bag. At the time of necropsy, storage stability spike standards were prepared in headspace vials using control blood (from untreated rats) spiked with known quantities of 1, 1, 2 Trichloroethane. In addition, unspiked control blood (from untreated rats) was frozen and shipped to Battelle Northwest Laboratories, Richland, Washington, for preparation of standards (for calibration curves) and samples of 1, 1, 2 Trichloroethane solution used to prepare the storage spike samples included. Plasma was prepared and frozen at 70° C. The plasma samples were shipped in Styrofoam containers on dry ice via overnight courier to Battelle Warehouse, Richland, Washington. The analytical laboratory and/ or the sponsor are responsible for subsequent data analysis and reporting of the pharmacokinetic evaluations. Individual dispositions and body weights for the pharmacokinetic animals are presented in Appendix I.

5.6. OPHTHALMIC EXAMINATIONS (TOXICOLOGY GROUPS)

Ocular examinations were conducted on all animals prior to the initiation of dosing (May 23,200l; study week l), near the end of the dosing period (August 23,200l; study week 12). All ocular examinations were conducted using an indirect ophthalmoscope (or other suitable equivalent equipment), preceded by pupillary dilation with an appropriate mydriatic agent. Examinations were performed by David A. Wilkie, D. V. M., M. S.,

D. A. C. V. O. (Appendix J).

5.7. ANATOMIC PATHOLOGY (TOXICOLOGY GROUPS)

5.7.1. MACROSCOPIC EXAMINATION A complete necropsy was conducted on all animals. Animals were euthanized by isoflurane anesthesia followed by exsanguination. The necropsies included, but were not limited to, examination of the external surface, all orifices, and the cranial, thoracic,

25 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane abdominal and pelvic cavities including viscera. The following tissues and organs were collected and placed in 10% neutral buffered formalin (except as noted):

Adrenals (2) Aorta Bone with marrow Sternum Bone marrow smear (taken from femur)* Brain (forebrain, midbrain, hindbrain) Epididymidesb Eyes with optic nerve (2) ' Exorbital lacrimal glands (2) Gastrointestinal tract Esophagus Stomach Duodenum Jej unum Ileum Cecum Colon Rectum Harderian glands (2) Heart Kidneys (2) Larynx (3 levels) d Liver (sections of two lobes) Lungs (including bronchi, fixed by inflation with fixative) Lymph nodes Mediastinal Tracheobronchial Mamrnary gland (females only) Nasal Tissuese Ovaries with oviducts (2) Pancreas Parathyroidsf Peripheral nerve (sciatic) Pituitary Prostate Salivary glands [mandibular (2)] Seminal vesicles (2) Skeletal muscle (rectus femoris) Skin Spinal cord (cervical, thoracic, lumbar) Spleen Testes (2) b Thymus Thyroids Trachea Urinary bladder Uterus with vagina All gross lesions (when possible)

a = Bone marrow smears were obtained at necropsy but not placed in formalin. b = Fixed in Bouin's solution. ' = Fixed in Davidson's solution. d = Three cross sections were examined in accordance with the method described by Renne4. e = The entire head was removed and preserved. Following decalcification, six cross sections of the nasal cavities were prepared for microscopic examination in accordance with the method described by Morgan'. f = Parathyroid glands were examined microscopically if in the plane of section and in all cases when a gross lesion was noted.

26 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 5.7.2. ORGAN WEIGHTS The following organs were weighed from all animals at the scheduled necropsies:

Adrenals Brain Epididymides Heart Kidneys Liver Lungs (prior to inflation with fixative) Ovaries Spleen Testes Thymus Thyroids (with parathyroids)"

a = Weighed after fixation.

Paired organs were weighed together. Organ to final body weight and organ to brain weight ratios were calculated.

5.7.3. SLIDEPREPARATION AND MICROSCOPICEXAMINATION Following fixation, protocol specified tissues were shipped to Experimental Pathology Laboratories, Inc. (EPL), Herndon, Virginia, for preparation of blocks and slides. EPL is responsible for the GLP compliance of the work conducted at their facility.

Microscopic examination was performed on all tissues listed in Section 5.7.1. from all animals in the control and 100 ppm groups at the scheduled necropsies. The larynx,

kidneys, liver, lungs, nasal tissues, trachea, and all gross lesions were examined from all animals in the 15 and 40 ppm groups at the primary necropsy. Missing tissues were identified as not found at necropsy, lost at necropsy, lost during processing, not in plane of section or other reasons as appropriate. Microscopic examinations were conducted by Frederick A. Ruecker, D. V. M., M. S., D. A. C. V. P., Experimental Pathology Laboratories,

Inc. (Appendix K).

5.8. STATISTICALMETHODS All analyses were conducted using two tailed tests for minimum significance levels of 1% and 5%, comparing each test article treated group to the control group by sex. Each mean was presented with the standard deviation (S. D.) and the number of animals (N)

27 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane used to calculate the mean. Statistical analyses were not conducted if the number of animals was two or less.

All statistical tests were performed using appropriate computing devices or programs.

Body weight, body weight change, food consumption, clinical pathology and organ weight data were subjected to a parametric one way analysis of variance (ANOVA) to determine intergroup differences. If the ANOVA revealed statistical significance (p< O. O5), Dunnett's test7 was used to compare the test article treated groups to the control group. Clinical pathology values for white blood cell types that occur at a low incidence (i. e., monocytes, eosinophils and basophils) were not subjected to statistical analysis.

28 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 6. RESULTS AND DISCUSSION (TOXICOLOGY GROUPS)

6.1. CLINICAL OBSERVATIONS AND SURVIVAL Summary Data: Tables 1,2,3,4,5 Individual Data: Table 20, Appendix F One female (no. 69377) in the 15 ppm group was euthanized in extremis on study day 30.

Clinical findings noted prior to euthanasia consisted of impaired use of hindlimbs, dried red material around both eyes, red discharge from both eyes and decreased defecation.

These findings were not considered test article related since they were not noted in the 40 and 100 ppm groups.

All other animals survived to the scheduled necropsy (study week 13). No test article related clinical signs were noted during the study. Clinical signs observed in the test article treated groups were noted similarly in the control group, were limited to single occasions for one animal, were not present in a dose related manner and/ or were those types of findings commonly observed in laboratory rats of this strain and age.

6.2. BODY WEIGHTS Summary Data: Tables 6, 7, 7A Individual Data: Tables 2 1,22 Test article related decreased mean body weights and body weight gains were noted during study week 0 to 1 in the 100 ppm group females. These differences were statistically significant (`~~ 0.05 or p< O. Ol) when compared to the control group.

There were no other test article related effects on body weights. However, several statistically significant (~~ 0.05 or p< O. Ol) differences in absolute body weight and body weight gains were noted when compared to the control group. Decreased mean body weights were noted in the 15 and 100 ppm group females at study week 7. Increased mean body weight gains were noted in the 15 and 100 ppm group males during study week 1 to 2, in the 15, 40 and 100 ppm group females during study week 7 to 8 and 29 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane decreased body weight gains were noted in the 15 and 100 ppm group females during study week 6 to 7. Since there were no time or dose related trends apparent, these differences were not test article related. No other remarkable differences were noted.

6.3. FOODCONSUMPTION Summary Data: Table 8 Individual Data: Table 23 There were no test article related effects on food consumption. However, several statistically significant (+ O. OS or ~~ 0.01) differences in food consumption were noted when compared to the control group. Increased mean food consumption was noted in the 15 ppm group males during study week 1 to 2 and in the 15 and 100 ppm group males during study weeks 8 to 10. Decreased food consumption was noted in the 40 and 100 ppm group females during study week 0 to 1, the 100 ppm group females during study week 6 to 7 and in the 15 and 100 group females during study week 12 to 13. Since there were no time or dose related trends apparent and/ or the differences from the control group were very small, these differences were not test article related. No other remarkable differences were noted.

6.4. CLINICALPATHOLOGY 6.4.1. HEMATOLOGY Summary Data: Tables 9, 10, 11 Individual Data: Tables 24,25,26 There were no test article related effects on hematology parameters.

30 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 6.4.2. SERUM CHEMISTRY Summary Data: Table 12 Individual Data: Table 27 Possible test article related increased mean cholesterol was noted in the 100 ppm group males and 40 and 100 ppm group females (statistically significant at ~~ 0.01 or ~~ 0.05 when compared to the control group). Mean glucose was decreased and statistically significant (pcO. 05) in the 100 ppm group females when the control and treated groups were compared. The difference from the control group appeared to be test article related for females exposed to the high concentration. There were no other potential test article

related effects on serum chemistry parameters.

6.5. OPHTHALMIC EXAMINATIONS Summary Data: Tables 13, 14, Appendix G Individual Data: Tables 28,29 No treatment related ophthalmic findings were noted at the study week 12 examination.

6.6. ANATOMIC PATHOLOGY 6.6.1. MACROSCOPIC EXAMINATION Summary Data: Tables 15, 16 Individual Data: Tables 30,3 1 There were no treatment related macroscopic findings at the scheduled necropsy or for the animal euthanized in extremis. Findings in the test article treated groups were noted similarly in the control group, were noted at a low incidence, typically single animals and/ or were findings common in laboratory rats.

31 of 619 1, 1,2 Trichloroethane 6.6.2. ORGAN WEIGHTS Summary Data: Tables 17, 18, 19 Individual Data: Tables 32,33,34 There were no test article related effects on organ weights. However, several statistically significant (~~ 0.05 or pcO. 01) differences were noted when the control and treated groups were compared. Mean kidney and heart weights relative to final body weight were increased in the 100 ppm group females and mean adrenal weights relative to final body weight and relative to brain weight were increased in the 15 ppm group females.

Since the absolute organ weights were not affected and no dose related trends were apparent, these differences were not considered test article related.

6.6.3. MICROSCOPIC EXAMINATION Summary Data: Appendix K Individual Data: Appendix K Test article related effects were noted in the liver and nasal turbinates of all test article

treated groups. Minimal hepatocellular vacuolization was noted in 3/ 10 females each from the 15 and 40 ppm groups and 5/ l 0 males and lO/ lO females from the 100 ppm group. The lesion was characterized by the presence of small vacuoles within the cytoplasm of the hepatocytes in a focal to multifocal distribution within the sections examined. In the females, there was no apparent dose related increase in severity between the three exposure groups, nor any dose related increase in incidence between the 15 and 40 ppm groups. While special stains were not employed to identify the material in the vacuoles, their morphology was consistent with lipid accumulation.

However, after 13 weeks of exposure to concentrations up to 100 ppm there were no microscopic findings indicative of centrilobular hepatocellular necrosis, the primary and most toxicologically significant effect on the liver of exposure to chlorinated short chain hydrocarbons, including 1,1,2 TCE. Centrilobular hepatocellular necrosis was observed in rats following a single exposure to 1,1,2 TCE at concentrations from 5 8 to 1527 ppm'.

32 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane Similarly, there were no test article related increases in serum levels of enzymes known to be increased following hepatocellular injury (e. g. increased levels of serum alanine aminotransferase).

Several microscopic lesions were noted in the nasal turbinates in the 40 and 100 ppm group males and all test article treated groups of females. The incidences of these lesions are delineated in the following text table:

Text Table 1: Incidence of Selected Microscopic Lesions in Nasal Turbinates Group:

Atrophy, Olfactory Epithelium VacuolizationiCyst, Olfactory Epithelium Respiratory Epithelium Metaplasia, Olfactory Epithelium 1 0 ppm M F O/ loa o/ 10 (O/ 60) b (O/ 60)

l/ IO l/ l0 (l/ 60) (1160)

o/ 10 o/ 10 (O/ 60) (O/ 60) 15 ppm M F o/ 10 o/ 10 (0160) (OW 2110 4110 (2/ 60) (4/ 60)

l/ l0 o/ 10 (1160) (0160)

" = Number of animals affected/ number of animals examined 3 40 ppm M F 6110 7/ 10 (S/ 60) (7/ 60)

6tlO 4/ 10 (11160) (4160)

l/ l0 l/ 10 (2/ 60) (l/ 60) 4 100 ppm M F 7110 lo/ lo (15/ 60) (13160)

lo/ lo 8/ 10 (E/ 60) (12/ 60)

3110 5/ 10 (3/ 60) (5160)

b = Total number of individual sections affected/ total number of individual sections examined Please note: bolded numbers are test article related Atrophy of the olfactory epithelium was characterized by a thinning of the olfactory epithelial mucosa primarily due to a reduction in the thickness of the nuclear cell layer.

The lesions were generally observed in sections from Levels 3, 4, and 5, of minimal to mild severity, and focal in distribution, primarily affecting the epithelium of the nasal septum in the dorsal medial meatus. Occasionally, there was a complete absence of the olfactory mucosa in the affected area. The mechanism for the cell loss was not microscopically obvious; however, since the lesion was only observed in animals from the 40 and 100 ppm groups, it was interpreted as related to test article exposure. The focal nature of the lesion could not be explained.

Vacuolization and/ or microcyst formation in the olfactory epithelium was characterized by the presence of small focal to multifocal vacuoles/ microcysts within the basilar regions of the olfactory mucosa. Again, the most commonly affected area was the 33 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane olfactory epithelium lining the dorsal medial meatus at Levels 2 through 5. Typically, the wall of these vacuoles/ cysts was devoid of any structure, but occasionally, they were lined by a cuboidal epithelium, which most resembled the epithelium of Bowman's glands. In general, the vacuoles were devoid of any content; however, occasionally an eosinophilic globular material was observed within. While the change was also observed in one control group male and female, the incidence of animals affected and levels per animal affected generally increased with exposure to increasing concentrations to the test article. In the low concentration group, the incidence of this finding was low (2 of 10 males and 4 of 10 females) and was observed in only one nasal section (i. e, level) per animal. Although test article related, this effect is considered to represent a slight response in the low exposure group. Although the pathogenesis of the lesion was not apparent, the character of the lesion suggests that it is probably degenerative in nature.

Whether exposure to the test article directly induced the formation of these vacuoles/ cysts or secondarily accelerated a process that is normally present within the nasal olfactory mucosa could not be determined.

Respiratory epithelial metaplasia of the olfactory epithelium was observed in males from the 15,40 and 100 ppm groups and females from the 40 and 100 ppm groups. This lesion was characterized by the presence of focal areas wherein the olfactory epithelium underwent metaplastic change to respiratory epithelium. Again, these changes were only observed in the olfactory epithelium lining the dorsal medial meatus at Levels 4 and 5.

Although one male in the 15 ppm group (no. 69335) also was affected, the lesion in this animal was considered incidental and not related to test article exposure since it was associated with inflammation within the nasal cavity interpreted as spontaneous in origin.

Respiratory epithelial metaplasia is commonly considered to be a sequela to atrophy and degeneration of the olfactory epithelium with replacement by ciliated and non ciliated respiratory epithelium. It may be seen as a spontaneous focal lesion in aged rats'.

Whether test article exposure directly induced this change or accelerated the normal aging process could not be determined.

34of619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane Additional microscopic findings (described below) noted in rats from control and test article treated groups included mononuclear cell infiltrates in various tissues,

granulomatous inflammation in the larynx and trachea, and mineralization or mineral deposition in the kidney, larynx, nasal turbinates, and cornea of the eye. In general, there were no significant differences in the incidence of these lesions between control and test article treated groups. Therefore, these findings were considered incidental and unrelated to test article exposure. Since the test article exposure was via inhalation, it should be emphasized that no test article related effects were observed for respiratory tract tissues other than the nasal tissues.

Mononuclear cell infiltrates, observed in various tissues, typically consisted of foci of lymphocytes and plasma cells generally accompanied by a few neutrophils as a minor component. This morphology was consistent with subacute inflammation.

Granulomatous inflammation was observed in the larynx at all three levels examined.

The lesion was characterized by the presence of histiocytes and an occasional multinucleated giant cell forming discrete focal microgranulomas surrounding a mineralized center located just below the mucosal epithelial layer. Similar lesions were also occasionally observed in the trachea. This lesion is typically observed as a response to a penetrating foreign body, and its location suggests that it may have resulted from the penetration of food material aspirated into the larynx and trachea. However, any such material, if present in the mineralized core, could not be identified.

Mineralization in the kidney of both control and treated male and female rats was characterized by the accumulation of lightly to intensely staining basophilic material within the lurnina of the renal tubules. The mineral deposition observed in the larynx was located within the central core of the microgranulomas, as already discussed. These changes were considered incidental and not related to treatment.

Mineral deposition in the nasal turbinates was observed in sections from Levels 2 through 6 of both control and test article treated male and female rats. This lesion was 35 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane characterized by the deposition of basophilic material just underlying the epithelium of the transitional, respiratory, and olfactory mucosae. In some cases, the mineral could be localized to the basement membrane. Similar deposits associated with the basement membrane of the respiratory and olfactory mucosae have been reported to increase in frequency with age5.

Mineral deposition within the cornea of the eye was characterized by the presence of lightly staining basophilic material within lamina propria of the cornea just underlying the cornea1 epithelium. The lesion was focal in distribution, typically located in the central region of the cornea, and of minimal severity. The pathogenesis of this lesion was not determined. However, since the incidence was not significantly different between controls and test article treated rats, it was considered incidental and not related to treatment.

Other findings in the test article treated groups were noted similarly in the control group,

were noted at a low incidence, typically single animals and/ or were findings common in laboratory rats.

36 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 7. CONCLUSIONS There were no test article related deaths. Clinical observations, body weights, food consumption, hematology parameters and organ weights were unaffected by test article exposure. No test article related ophthalmic findings were noted, and no test article

related macroscopic changes were observed at the scheduled necropsy.

Test article related effects noted in the 15 ppm group consisted of:

l Minimal hepatocellular vacuolization in 3/ l 0 females.

l Microscopic lesions in the olfactory epithelium of the nasal turbinates (vacuolizatiotimicrocyst formation in 2/ l 0 males and 4/ l 0 females).

Test article related effects noted in the 40 ppm group consisted of:

l Minimal hepatocellular vacuolization in 3/ l 0 females.

l Microscopic lesions in the olfactory epithelium of the nasal turbinates (atrophy in 6/ 10 males and 7/ l 0 females, vacuolization/ microcyst formation in 6/ l 0 males and 4/ 10 females and respiratory epithelial metaplasia in l/ 10 males and l/ 10 females).

Possible test article related effects noted in the 100 ppm group consisted of:

l Decreased absolute body weights and body weight gains from study week 0 to 1 in females.

l Increased mean cholesterol levels in the males and females and decreased mean glucose in females.

l Minimal hepatocellular vacuolization in 5/ 10 males and lo/ 10 females.

l Microscopic lesions in the olfactory epithelium of the nasal turbinates (atrophy in 7/ l 0 males and lo/ 10 females, vacuolization/ microcyst formation in lO/ lO males and S/ 10 females and respiratory epithelial metaplasia in 3/ 10 males and 5/ 10 females).

Chlorinated, short chain hydrocarbons are believed to be activated to oxidative species by cytochrome P450 metabolizing enzymes of hepatocytes in the centrilobular regions of the liver. Following 13 weeks of inhalation exposure to 1,1,2 TCE, test article related microscopic findings in the liver were limited to minimal hepatocellular vacuolization,

which might represent lipid accumulation. However, the presence of this finding without 37 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane centrilobular necrosis or an increase in serum levels of enzymes indicative of hepatocellular injury suggests that this minimal degree of vacuolization (in particular where only 3 of 10 females were affected) lacks toxicological significance'. It is possible that the rate of oxidative events is too slow at these exposure levels to produce frank cellular injury or the hepatocytes have become insensitive to such events.

A relatively high concentration of microsomal metabolizing enzymes is present in the rodent nasal olfactory epithelium. As a result, the nasal olfactory epithelium of the rat is extremely sensitive to agents that may be activated by cytochrome P 450 metabolizing enzymes. It has been reported that the fraction of the nasal airways covered by olfactory epithelium is approximately 50% in the F344 rat compared to 3% in the human". In addition, based on information cited by Haschek and Witschi, the relative surface area of the nasal airways (normalized to body weight) is approximately 16 times greater in the rat compared to the human' I. Therefore, the rat may be considered one of the most sensitive animal models for chemical injury to the olfactory epithelium. Olfactory epithelial necrosis was observed in rats following a single exposure to 1,1,2 TCE in an earlier study'. Following the subchronic exposures in the current study, animals in the high and mid concentration groups had changes in the olfactory epithelium, including atrophy and respiratory epithelial metaplasia. A third test article related finding in the olfactory epithelium, vacuolizationimicrocyst formation, was observed for animals in all 1,1,2

TCE exposed groups. However, the slight response observed in the low concentration group was not considered to represent an adverse response since the finding was not accompanied by frank cellular injury (i. e., necrosis, atrophy or metaplasia). Since the test article exposure was via inhalation, it should also be emphasized that no test article

related effects were observed for respiratory tract tissues other than the nasal tissues.

Based on the results of this study, the no observed effect level (NOEL) for whole body inhalation of 1, 1, 2 Trichloroethane to rats for 13 weeks was less than 15 ppm.

However, the no observed adverse effect level (NOAEL) was less than 40 ppm since the hepatic and nasal effects observed for the low concentration group were not considered 38 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane toxicologically significant. In addition, a major difference in chemical sensitivity of the nasal airways between the rat and human may exist because of the vastly greater presence of olfactory epithelium in the rat. Therefore, the presence of atrophy and metaplasia in a tissue that is extremely sensitive in the rat compared to other species may be misleading in estimating human risk.

39of619 WEL 417002 HAP Task Force I ) 1,2 Trichloroethane 8. KEYSTUDYPERSONNELANDREPORTSUBMISSION Study Supervisors:

Susan C. Haley, B. S. Jonathan M. Hurley, B. S. Sally A. Keets, A. S. Carol A. Kopp. B. S.

Michael A. Safron, AS., H. T. (A. S. C. P.) Daniel W. Sved, Ph. D., Acting Group Supervisor, Clinical Pathology Operations Manager, Genera! Toxicology Acting Senior Operations Manager, Vivarium Manager, Gross Pathology and Developmental Toxicology Laboratory Manager, Histology Director, MetaboIism and Analytical Chemistry Report Prepared By: `A Study Analyst Reviewed By:

Robert A. W" ally, B. S# Group Supervisor, Study A. nalysi& d Reports Director, Inhalati6n Toxicology 40 of 619 WIL 4 17002 HAP Task Force 9. QUALITYASSURANCE UNITSTATEMENT 9.1. PHASESINSPECTED Date( s) of Insnection( s) 6/ 20/ O 1 612710 1 7/ l l/ O1 713 o/ o 1 x/ 30/ 0 1 1 l/ 15/ 01 11115, 16/ 01 1 l/ 16,26/ 01 1 l/ 16/ 01 1 l/ 16/ 01 11/ 28,29/ 01, 1131, 217102 1 l/ 28,29/ 01, 2/ 4,7/ 02 1 l/ 28,29/ 01, 2/ 1,7/ 02 12/ 27/ 01, l/ 30, 217102 l/ 30,2/ 7/ 02 211,3,7, X102 217102 217102 1120,22,24,25, 28102,2/ 4/ 02 Phase Insnected Test Article Exposure Blood Collection Detailed Physicals/ Body WeightslFood Weights Animal Care and Equipment Necropsy Study Records (I l) Study Records (I 2) Study Records (I 3) Study Records (N 1) Study Records (C 1,2,3) Date( s) Findings Reported to Studv Director 6/ 2010 1 612710 1 Date( s) Findings Reported to Management 7/ 30/ 01 7130/ 01 7/ l l/ O1 713 l/ O1 813010 1 1 l/ 26/ 01 1112610 1 1 l/ 29/ 01 1 l/ 26/ 01 1 l/ 2910 1 8/ 3 l/ O1 Xl3 l/ 01 9128/ O 1 12/ 26/ 01 12/ 26/ O 1 12/ 26101 12/ 26/ O 1 12126101 Study Records (Ex 3) 217102 3128102 Study Records (Ex 4) 217102 3128102 Study Records (Ex 5) 217102 3128102 Study Records (Ex 1) 217102 3128102 Study Records (Ex 2) 217102 3128102 Draft Report (Inhalation) 218102 3128102 Study Records (A l) 217102 3128102 Draft Report (Purity Assessment) 218102 3128102 Draft Report (w/ o Inhalation and Purity Reports) 215102 3 /28/ 02 1, 1,2 Trichloroethane This study was inspected in accordance with the U. S. Environmental Protection Agency TSCA guideline 799.9346: 90 Day Inhalation Toxicity Study, the standard operating procedures of WIL Research Laboratories, Inc., and the sponsor's protocol and protocol amendments with the following exceptions. The data located in Appendix A (Certificate of Analysis) were the responsibility of the sponsor. The data located in Appendix F (Serological Screening) were the responsibility of BioReliance Corporation. The data located in Appendix K (Histopathology Report) were the responsibility of Experimental Pathology Laboratories, Inc. Quality Assurance findings, derived from the inspections during the conduct of the study and from the inspections of the raw data and draft report,

41 of 619 1, 1,2 Trichloroethane are documented and have been reported to the study director. A status report is submitted to management monthly.

This report accurately reflects the data generated during the study. The methods and procedures used in the study were those specified in the protocol, its amendments and the standard operating procedures of WIL Research Laboratories, Inc.

42 of 619 WIL4 17002 HAP Task Force 1, 1,2 Trichloroethane 9.2. APPROVAL This study was inspected according to the criteria discussed in Section 9.1.

ReDot t Audited BY:

F& byn N. Durr, B. S. Auditor I, Quality Assurance Report Reviewed BY:

Margaret A. Franklin Unit Supervisor, Quality Assurance a Date 43 of 619 WIL 417002 HAP Task Force 1, 1,2 Trichloroethane 10. REFERENCES 1. WIL 417001. (2001) Acute Inhalation Toxicity (with Histopathology) Study of 1,1,2 Trichloroethane (1,1,2 TCE) in Rats. WIL Research Laboratories, Inc., Ashland, OH.

2. Torkelson, T. R. (1994) Halogenated aliphatic hydrocarbons containing chlorine, bromine, and iodine. In: Patty's Industrial Hygiene and Toxicology, Fourth Edition (Clayton, G. D. and Clayton, F. E., eds.) John Wiley & Sons, Inc., New York, NY., pp. 41284132.

3. National Resource Council (1996) Guide for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, Commission on Life Sciences. National Academy Press, Washington, DC.

4. Snedecor, G. W. and Co& ran, W. G. (1980) One way classifications; analysis of variance. In: Statistical Methods, Seventh Edition. The Iowa State University Press, Ames, IA, pp. 215 237.

5. Dunnett, C. W. (1964) New tables for multiple comparisons with a control. Biometrics, 20: 482 49 1,

6. Renne, R. A., Gideon, K. M., Miller, R. A., Mellnick, P. W., and Grumbein, S. L. (1992). Histologic methods and interspecies variations in the laryngeal histology of F344/ N rats and B6C3Fl mice. Toxicologic Pathology 20( 1); 44 5 1.

7. Morgan, K. T. (199 1) Approaches to the identification and recording of nasal lesions in toxicology studies. Toxicologic Pathology, 19( 4): 337 351.

8. Greaves, P. (2000) Histopathology of Preclinical Toxicity Studies: Interpretation and Relevance in Drug Safety Evaluation, Second Edition. Elsevier Science, Amsterdam, The Netherlands, pg. 2 11.

9. Cattley, R. C. and Popp, J. A. (2002) Liver. In: Handbook of Toxicologic Pathology, Second Edition, (Haschek, W. M., Rousseaux, C. G. and Wallig, M. A., eds.) Academic Press, San Diego, CA, pp. 187 225.

10. Dungworth, D. L., Hahn F. F. and Nikula, K. J. (1995) Noncarcinogenic responses of the respiratory tract to inhaled toxicants. In: Concepts in Inhalation Toxicology, Second Edition, (McClellan, R. O. and Henderson, R. F., eds.) Taylor & Francis, Washington, D. C., pp. 533 576.

44of619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane 11. Haschek, W. M and Wits& i, H. R. (1991) Respiratory system. In: Handbook of Toxicologic Pathology (Haschek, .M. and Rousseaux, C. G., eds.) Academic Press, Inc., San Diego, CA, pp. 761 827.

45 of 619 WIL 4 17002 Hap Task Force 1, 1,2 Trichloroethane 11. DEVIATIONS FROM THE PROTOCOL This study was conducted in accordance with the protocol and protocol amendments,

except for the following.

l Section 4.1.2. On July 26, 2001, the Group 1 (0 ppm) animals were inadvertently placed in the 15 ppm exposure chamber and the Group 2 (15 ppm) animals were inadvertently placed in the 0 ppm exposure chamber for the six hour exposure period.

l Section 4.1.2. The negative pressure and chamber ventilation rates for chambers l 4 were not recorded at one interval on August 3, 2001; therefore, the mean negative pressure and CVR were calculated from nine readings.

l Section 4.1.2. The mean chamber ventilation rate for chamber 3 descended below the stated range of 400 500 LPM on August 14,28 and 30,200l.

l Section 4.1.3. Eight mm glass beads were also used in the vaporization column.

l Section 4.1.4. The standards prepared on July 26, 2001 and August 16, 2001 were out of the + 10% range as specified by the protocol. The low standard (9 ppm), prepared on July 26,2001, was approved by the study director for a range of + 15%.

l Section 4.1.5. On July 27, 2001, chamber 1 was exposed for only four and one half hours. This exposure yielded nine environmental samples and eight concentration samples.

l Section 5.7. On August 30,2001, the heart for animal no. 69346 (Group 4 male) was lost at necropsy prior to weighing. Therefore, a weight was not recorded and histopathological examination was not possible.

These deviations did not negatively impact the quality or integrity of the data nor the outcome of the study.

46 of 619 WIL 4 17002 HAP Task Force 1, 1,2 Trichloroethane TABLES 1  31 47 of 619 m II e r 4 48 of 619 TABLE 1 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 2 SPONSOR: HAP TASK FORCE SUMMARY OF SURVIVAL AND DISPOSITION FEMALES GROUP : 1 2 3 4 ______________________________________ WEEK LIVE FD EE SE LIVE FD EE SE LIVE FD EE SE LIVE FD EE SE 0 10 0 0 0 10 0 0 0 10 0 0 0 10 0 0 0 1 10 0 0 0 10 0 0 0 10 0 0 0 10 0 0 0 2 10 0 0 0 10 0 0 0 10 0 0 0 10 0 0 0 3 10 0 0 0 10 0 0 0 10 0 0 0 10 0 0 0 4 10 0 0 0 9 010 10 0 0 0 10 0 0 0 5 10 0 0 0 9 0 0 0 10 0 0 0 10 0 0 0 6 10 0 0 0 9 0 0 0 10 0 0 0 10 0 0 0 7 10 0 0 0 9 0 0 0 10 0 0 0 10 0 0 0 a 10 0 0 0 9 0 0 0 10 0 0 0 10 0 0 0 9 10 0 0 0 9 0 0 0 10 0 0 0 la 0 0 0 10 10 0 0 0 9 0 0 0 10 0 0 0 10 0 0 0 11 10 0 0 0 9 0 0 0 10 0 0 0 10 0 0 0 12 10 0 0 0 9 0 0 0 10 0 0 0 10 0 0 0 _'"_____"__"__"_ '"__________________"__"~~~~~~~~~~~~~~~~~~

WEEK = WEEK OF STUDY FD = FOUND DEAD EE = EUTHANIZED IN EXTREMIS SE = SCHEDULED EUTHANASIA ______________________________________

l 0 PPM 2 15 PPM 3 40 PPM 4 100 PPM PSURVv4.02 1x/ 28/ 2001 TABLE 2 (DETAILED PHYSICAL EXAMINATIONS/ DISPOSITIONS) PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 1 SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS  MALE ___ ______________________________________ TABLE RANGE: 05 31 01 TO 08 31 01 GROUP: 1 2 3 4 ______________________________________

NORMAL NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION PRIMARY NECROPSY (WEEK 13)

EYES/ EARS/ NOSE DRIED RED MATERIAL AROUND RIGHT EYE DRIED RED MATERIAL AROUND LEFT EYE RED DISCHARGE RIGHT EYE RED DISCHARGE LEFT EYE OPACITY LEFT EYE EXCRETA DECREASED DEFECATION 0" _ SOFT FECES % o\ BODY/ INTEG II z SCABBING END OF TAIL ORAL/ DENTAL UPPER INCISOR( S) MALALIGNED _______________________________ l 0 PPM 2 15 PPM 115/ 10 lO/ lO I/ 1 4/ 3 1/ 1 2/ 2 1/ 1 1/ 1 1/ 1 o/ 0 o/ 0 _______________________________ 3 40 PPM 4 100 PPM 123/ 10 118/ 10 124/ 10 lO/ lO 1/ 1 3/ 2 o/ 0 3/ 3 o/ 0 o/ 0 1/ 1 o/ 0 o/ 0 ___________________ lO/ lO lO/ lO 01 0 1/ 1 4/ 2 o/ 0 o/ 0 I/ 1 2/ 1 o/ 0 :: f 3/ 1 o/ 0 2/ 1 o/ 0 51 of 619 TABLE 2 (DETAILED PHYSICAL EXAMINATIONS~ DISPOSITI~ N~) PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 3 SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS _____ F E M A L E _____ ______________________________________

TABLE RANGE: 05 31 01 TO 08 31 01 GROUP: 1 2 3 4 ________________________________________

NORMAL NO SIGNIFICANT CLINICAL OBSERVATIONS 102/ 10 92/ 10 104/ 10 lOP/ lO DISPOSITION EUTHANIZED IN EXTREMIS DUE PHYSICAL Q/ 0 1/ 1 o/ 0 Q/ 0 CONDITION PRIMARY NECROPSY (WEEK 13) lO/ lO g/ 9 lO/ lO 10/ 3.0 BEHAVIOR/ CNS IMPAIRED USE OF LEFT HINDLIMB Q/ 0 11 1 o/ 0 o/ 0 IMPAIRED USE OF RIGHT HINDLIMB Q/ 0 1/ 1 o/ 0 Q/ 0 CARDIO PULMONARY RALES Q/ 0 Q/ 0 2/ 2 1/ 1 EYES/ EARS/ NOSE DRIED RED MATERIAL AROUND RIGHT EYE Q/ 0 1/ 1 o/ 0 o/ 0 DRIED RED MATERIAL AROUND LEFT EYE 2/ 2 2/ 2 2/ 1 Q/ 0 RED DISCHARGE RIGHT EYE I/ J. 1/ 1 o/ 0 o/ 0 RED DISCHARGE LEFT EYE o/ cl 2/ 2 o/ 0 o/ 0 OPACITY RIGHT EYE 3/ 1 Q/ 0 Q/ 0 Q/ 0 PUPIL DILATED RIGHT EYE l/ 1 o/ 0 o/ 0 Q/ 0 ________________________________________

l 0 PPM 2 15 PPM 3 40 PPM 4 100 PPM TABLE 2 (DETAILED PHYSICAL EXAMINATIONS/ DISPOSITI~ NS) PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 4 SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS  F E M A L E _ __________________~

TABLE RANGE: 05 31 01 TO 08 31 01 GROUP: 1 2 3 4 _______________

EXCRETA DECREASED DEFECATION o/ 0 1/ 1 o/ 0 o/ 0 SOFT FECES o/ 0 o/ 0 o/ 0 I/ 1 BODY/ INTEG II SCABBING END OF TAIL o/ 0 41 1 2/ 1 o/ 0 SPECIAL I WET YELLOW MATERIAL UROGENITAL AREA 19/ 8 16/ 8 8/ 4 lO/ 5 DRIED YELLOW MATERIAL UROGENITAL AREA 5/ 3 4/ 2 12/ 6 v/ 5 DRIED YELLOW MATERIAL VENTRAL TRUNK o/ 0 o/ 0 1/ 1 o/ 0 WET YELLOW MATERIAL DORSAL TRUNK o/ 0 1/ 1 o/ 0 o/ 0 SPECIAL II 2 END OF TAIL MISSING o/ 0 lO/ 1 o/ 0 o/ 0 TAIL BROKEN o/ 0 1/ 1 o/ 0 o/ 0 or ________________________________________ OY l 0 PPM 2 15 PPM 3 40 PPM 4 100 PPM s PCSUV4.03 02/ 06/ 2002 TABLE 3 (PRIOR ~0 DOSING) PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 1 SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS ____ MALE __ ________________________________________ TABLE RANGE: 05 31 01 TO 08 30 01 GROUP: 1 2 3 4 ________________________________________

EYES/ EARS/ NOSE DRIED RED MATERIAL AROUND NOSE o/ 0 DRIED RED MATERIAL AROUND RIGHT EYE 4/ 4 DRIED RED MATERIAL AROUND LEFT EYE 3/ 2 RED DISCHARGE RIGHT EYE I/ 1 RED DISCHARGE LEFT EYE 1/ 1 EXCRETA DECREASED DEFECATION o/ 0 SOFT FECES I/ 1 SPECIAL I WET YELLOW MATERIAL UROGENITAL AREA 2/ 1 DRIED YELLOW MATERIAL UROGENITAL AREA o/ 0 3 WET BROWN MATERIAL ANOGENITAL AREA o/ 0 _ _____ _________________ g l O PPM 2 15 PPM 3 40 PPM 4

0\

6 2/ 1 o/ 0 o/ 0 o/ 0 01 0 2/ 1 4/ 1 3/ 1 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 1/ 1 o/ 0 o/ 0 11 1 o/ 0 2/ 2 o/ 0 o/ 0 I/ 1 l/ 1 o/ 0 4/ 3 o/ 0 o/ 0 I/ 1 ____________________________________ 100 PPM PROJECT NO.: WIL 417002 TABLE 3 (PRIOR TO DOSING) 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 2 SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS _____ F E M A L E ____ __________________~~~ TABLE RANGE: 05 31 01 TO 08 30 01 GROUP: 1 2 3 4 __________

BODY/ INTEGUMENT DERMAL ATONIA l/ 1 EYES/ EARS/ NOSE DRIED RED MATERIAL AROUND RIGHT EYE 3/ 2 DRIED RED MATERIAL AROUND LEFT EYE 3/ 2 EXCRETA DECREASED DEFECATION SPECIAL I WET YELLOW MATERIAL UROGENITAL AREA DRIED YELLOW MATERIAL UROGENITAL AREA 13/ 6 7/ 6 DRIED YELLOW MATERIAL VENTRAL TRUNK o/ 0 s DRIED BROWN MATERIAL ANOGENITAL AREA o/ 0 WET YELLOW MATERIAL HINDLIMB 1/ 1 % WET YELLOW ANOGENITAL AREA 21 1 o PPM 2 15 PPM 3 40 PPM 4 100 PPM __ o/ 0 o/ 0 1/ 1 o/ 0 2/ 1 I/ 1 o/ 0 o/ 0 14/ 5 lO/ 4 9/ 4 4/ 3 11 1 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 _______ o/ 0 o/ 0 o/ 0 o/ 0 12/ 5 16/ 6 o/ 0 1/ 1 o/ 0 o/ 0 ________

PCSUv4.03 02/ 06/ 2002 R: 02/ 06/ 2002 TABLE 4 (FOLLOWING EXPOSURE) PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 1 SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS __ MALE ___ ________________________________________

TABLE RANGE: GROUP:

EYES/ EARS/ NOSE DRIED RED MATERIAL AROUND NOSE DRIED RED MATERIAL AROUND RIGHT EYE WET RED MATERIAL AROUND LEFT EYE DRIED RED MATERIAL AROUND LEFT EYE RED DISCHARGE RIGHT EYE RED DISCHARGE LEFT EYE CLEAR DISCHARGE RIGHT EYE CLEAR DISCHARGE LEFT EYE RED NASAL DISCHARGE EXCRETA SOFT FECES DERMAL OBS WET YELLOW MATERIAL UROGENITAL AREA DRIED YELLOW MATERIAL UROGENITAL AREA DRIED BROWN MATERIAL ANOGENITAL AREA WET YELLOW ANOGENITAL AREA _______________________________________

l o PPM 2 15 PPM 3 __ 1 _________ 05 31 01 TO 08 30 01 2 3 _________

I/ 1 o/ 0 o/ 0 53/ 2 o/ 0 2/ 1 o/ 0 1/ 1 o/ 0 o/ 0 o/ 0 o/ 0 2/ 1 o/ 0 2/ 2 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 01 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 4 __________________

o/ 0 8/ 2 o/ 0 1/ 1 4/ 1 o/ 0 1/ 1 o/ 0 o/ 0 o/ 0 4/ 2 4/ 2 o/ 0 01 0 40 PPM 4 100 PPM PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 4 (FOLLOWING EXPOSURE) 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS PAGE 2 TABLE RANGE: 05 31 01 TO 08 30 01 GROUP: 1 2 3 4 ________________________________________ __________________________________

BODY/ INTEGUMENT DERMAL ATONIA EYES/ EARS/ NOSE DRIED RED MATERIAL AROUND RIGHT EYE DRIED RED MATERIAL AROUND LEFT EYE RED DISCHARGE RIGHT EYE RED DISCHARGE LEFT EYE EXCRETA SOFT FECES ORAL/ DENTAL UPPER INCISOR( S) MALALIGNED 2 DERMAL OBS 0, WET YELLOW MATERIAL UROGENITAL AREA OY DRIED YELLOW MATERIAL UROGENITAL AREA ;; DRIED YELLOW MATERIAL VENTRAL TRUNK WET BROWN MATERIAL ANOGENITAL AREA WET YELLOW MATERIAL HINDLIMB WET YELLOW ANOGENITAL AREA 41 3 2/ 1 a/ 2 o/ 0 o/ 0 o/ 0 21/ 7 13/ 7 o/ 0 o/ 0 11 1 2/ 1 o/ 0 42/ 8 12/ 5 o/ 0 1/ 1 o/ 0 o/ 0 o/ 0 1/ 1 o/ 0 o/ 0 o/ 0 o/ 0 55/ 10 7/ 5 I/ 1 o/ 0 o/ 0 o/ cl ________________________________________ l 0 PPM 2 15 PPM 3 40 PPM 4 100 PPM PCSUV4.03 02/ 06/ 2002 R: 02/ 06/ 2002 TABLE 5 (NON EXPOSURE DAYS) PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS PAGE 1 __ MALE ____  _ _ _  _ _

TABLE RANGE: 05 31 01 TO 08 30 01 GROUP: 1 2 3 4 EYES/ EARS/ NOSE DRIED RED MATERIAL AROUND NOSE DRIED RED MATERIAL AROUND RIGHT EYE DRIED RED MATERIAL AROUND LEFT EYE RED DISCHARGE LEFT EYE EXCRETA SOFT FECES SPECIAL WET YELLOW MATERIAL UROGENITAL AREA DRIED YELLOW MATERIAL UROGENITAL AREA WET BROWN MATERIAL ANOGENITAL AREA DRIED BROWN MATERIAL ANOGENITAL AREA ___________________ kz 1 0 PPM 2 15 PPM 0, o\

s 1/ 1 o/ 0 2/ 2 o/ 0 1/ 1 1/ 1 o/ 0 2/ 2 o/ 0 1/ 1 o/ 0 1/ 1 2/ 2 o/ 0 2/ 1 o/ 0 1/ 1 o/ 0 1/ 1 o/ 0 _________________~~ 3 40 PPM 4 100 PPM 59 of 619 J 60of619 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 6 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF BODY WEIGHTS [G] PAGE 2 __ MALE ____ GROUP: 0 PPM 15 PPM 40 PPM 100 PPM _______________________________________

WEEK 3 MEAN 235. 243. 237. 239. S. D. 10.3 15.7 18.0 10.5 N 10 10 10 10 4 MEAN 248. 257. 250. 254. S. D. 11.4 15.9 21.4 11.3 N 10 10 10 9 5 MEAN 257. 267. 261. 265. S. D. 13.0 16.3 22.2 12.9 N 10 10 10 10 6 MEAN 268. 277. 270. 275.

S. D. N 15.6 10 20.0 10 22.1 10 12.8 10 7 MEAN 280. 290. 280. 284. S. D. 15.8 21.4 24.9 10.5 N 10 10 10 10 ________________________________________

None significantly different from control group TABLE 6 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 3 SPONSOR: HAP TASK FORCE SUMMARY OF BODY WEIGHTS tG1 ________________________________________

GROUP: ________________________________________

WEEK 8 MEAN S. D. N 9 MEAN S. D. N 10 MEAN S. D. N 11 s MEAN S. D.

0, N m 5; 12 MEAN S. D. N 0 PPM ______

285.

15.3 10 292. 306. 294. 302. 16.2 19.3 23.2 13.4 10 10 10 10 286. 300. 294. 304. 19.4 23.6 24.7 12.9 10 10 10 10 299. 320. 307. 317.

29.0 23.3 29.0 13.4 10 10 10 10 308. 326. 311. 322.

22.2 23.4 28.8 14.3 10 10 10 10 MALE ____ 15 PPM 300.

20.0 10 40 PPM 100 PPM 289. 294.

24.6 14.5 10 10 ________________________________________ None significantly different from control group 63 of 619 TABLE 6 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 5 SPONSOR: HAP TASK FORCE SUMMARY OF BODY WEIGHTS [Gl ___________________

_____ F E M A L E ___

GROUP: 0 PPM 15 PPM 40 PPM 100 PPM _______________________________________ WEEK 2 MEAN 95. 91. 98. 94. S. D. 8.1 9.5 9.6 1.7 N 10 10 10 10 1 MEAN 110. 108. 109. 107. S. D. 7.0 7.3 6.6 6.9 N 10 10 10 10 127. 128. 128. 127. 5.8 5.9 6.1 5.5 10 10 10 10 0 MEAN S. D. N 1 2 MEAN 145. 141. 140. S. D. 5.4 6.1 5.9 137.* 3.7 0 N 10 10 10 10 ch o\

G 2 MEAN 155. 152. 149. 148. S. D. 5.1 6.8 9.1 6.2 N 10 10 10 10 _____________________________________ * = Significantly different from the control group at 0.05 using Dunnett's test TABLE 6 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 6 SPONSOR: HAP TASK FORCE SUMMARY OF BODY WEIGHTS [G]

____________________________________ ____ F E M A L E _____ GROUP: 0 PPM 15 PPM 40 PPM 100 PPM ______________~~~ WEEK 3 MEAN 164. 159. 159. 157. S. D. 5.7 8.0 7.5 6.0 N 10 10 10 10 4 MEAN 171. 166. 167. 164. S. D. 5.8 7.9 9.0 8.2 N 10 10 10 9 5 MEAN 177. 172. 173. 171. S. D. 9.1 8.3 7.7 7.5 N 10 9 10 10 6 MEAN 182. 175. 175. 175. S. D. 8.5 8.1 9.1 8.5 N 10 9 10 10 7 MEAN 189, 178.* 180. 178.* S. D. 9.1 8.5 8.9 7.3 N 10 9 10 10 ________________ * = Significantly different from the control group at 0.05 using Dunnett's test PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 6 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF BODY WEIGHTS [G] PAGE 7 _______________________________________

_____ F E M A L E ____

GROUP: 0 PPM 15 PPM 40 PPM 100 PPM ______________________________________

WEEK 8 MEAN 190. 182. 185. 182. S. D. 10.1 8.5 9.9 6.7 N 10 9 10 10 9 MEAN S. D. N 10 MEAN S. D. N 11 MEAN S. D. N 12 MEAN S. D. N 194. 188. 188. 187. 11.7 8.7 11.5 7.9 10 9 10 10 191. 184. 188. 185. 7.4 9.7 10.3 9.4 10 9 10 10 197. 192. 195. 192. 20.3 9.8 10.8 7.8 10 9 10 10 202. 192. 197. 193. 13.5 10.1 11.9 9.6 10 9 10 10 ______________________________________   _ _ _  _

None significantly different from control group 67 of 619 I *

68 of 619 TABLE 7 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 2 SPONSOR: HAP TASK FORCE SUMMARY OF BODY WEIGHT CHANGES [G]

______________________________________ ___ MALE __ GROUP: 0 PPM 15 PPM 40 PPM 100 PPM ______________________________________ WEEK 3T0 4 MEAN 13. 15. 13. 14. S. D. 2.1 7.7 4.1 3.3 N 10 10 10 9 4T0 5 MEAN 9. 9. 10. 13. S. D. 6.1 4.7 4.0 3.7 N 10 10 10 9 5T0 6 MEAN 10. 11. 9. 10. S. D. 5.4 5.6 4.1 3.2 N 10 10 10 10 6T0 7 % MEAN 12. 12. 10. 9. S. D. 2.7 3.8 3.9 4.2 OH, N 10 10 10 10 OY G 7T0 MEAN 8 5. 10. 9. 10. S. D. 6.3 3.5 4.9 5.5 N 10 10 10 10 I I I I . .

70 of 619 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 7 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF BODY WEIGHT CHANGES [Gl PAGE 4 ________________________________~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ____ F E M A L E _____

GROUP: 0 PPM 15 PPM 40 PPM 100 PPM _______________________________________

WEEK 2 TO 1 MEAN 14. 11. S. D. 2.9 2.9 N 10 10 lTO 0 MEAN 18. 19. S. D. 3.9 3.2 N 10 10 OTO 1 MEAN 17. 14. S. D. 5.3 4.3 N 10 10 1TO 2 MEAN 11. 10. S. D. 3.2 3.9 N 10 I. 0 2T0 3 MEAN 9. 8. S. D. 2.7 1.9 N 10 10 ________________________________________

** = Significantly different from the control group at 0.01 using Dunnett's test 10. 13. 5.0 2.6 10 10 19. 20. 3.1 2.8 10 10 12. 5.4 10 9. 10. 4.3 4.8 10 10 10. 9. 3.8 3.0 10 10 lo.** 4.7 10 72 of 619 .mo d .r l N .mo r .r l cr 73 of 619 TABLE ?A PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SPONSOR: HAP TASK FORCE SUMMARY OF CUMULATIVE BODY WEIGHT CHANGES [Gl ________________________________________ _ MALE ____ GROUP: 0 PPM 15 PPM _______________________________________ WEEK OTO 1 MEAN 30. 32. S. D. 7.6 4.6 N 10 10 OTO 2 MEAN 48. 57. S. D. 10.9 8.3 N 10 10 OTO 3 MEAN 66. 73. S. D. 10.8 13.5 N 10 10 2 OTO 4 MEAN 79. 87. S. D. 12.2 14.2 0  i N 10 10 o\

5, OTO MEAN 5 88. 97. S. D. 13.9 14.8 N 10 10 40 PPM ___

27. 27. 7.1 7.6 10 10 49. 51. 8.4 9.9 10 10 67. 69. 11.6 12.4 10 10 80. 85. 14.8 12.7 10 9 91. 15.5 10 None significantly different from control group PAGE 1 _______________

100 PPM _________

95. 16.0 10 75 of 619 76 of 619 77 of 619 PROJECT NO.: WIL 417002 .SPONSOR: HAP TASK FORCE TABLE 7A 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF CUMULATIVE BODY WEIGHT CHANGES [Gl PAGE 5 _______________________________________

____ F E M A L E _____

GROUP: 0 PPM 15 PPM 40 PPM 100 PPM WEEK OTO 6 MEAN 55. 48. 48. 48. S. D. 11.9 10.2 11.0 12.3 N 10 9 10 10 OTO 7 MEAN 62. 50. 52. 50. S. D. 11.8 11.5 9.6 11.3 N 10 9 10 10 OTO 8 MEAN 63. 55. 57. 55. S. D. 13.7 11.1 10.9 10.4 N 10 9 10 10 OTO 9 2 MEAN 67. 60. 60. 59. S. D. 14.6 11.9 12.3 11.8 0 ++ J, N 10 9 10 10 6\

;;; OTO MEAN 10 63. 56. 60. 58. S. D. 10.4 13.2 11.4 12.4 N 10 9 10 10 _______________________________________ None significantly different from control group 79 of 619 80 of 619 TABLE 8 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 2 SPONSOR: HAP TASK FORCE SUMMARY OF WEEKLY ~00~ CONSUMPTION [G/ ANIMAL/ DAY]

_____________________________________ __ MALE __ GROUP: 0 PPM 15 PPM 40 PPM 100 PPM ________________________________________

WEEK 4T0 5 MEAN 17. 17. 17. 17. S. D. 1.2 0.9 1.1 0.9 N 10 10 10 10 5T0 6 MEAN 17. 18. 17. 17. S. D. 1.1 1.4 1.4 0.8 N 10 10 10 10 6T0 7 MEAN 17. 17. 16. 17. S. D. 1.0 1.4 1.3 0.9 N 10 10 10 10 7T0 8 MEAN 17. S. D. 0.9 N 10 18. 17. 17. 1.3 1.2 0.7 10 10 10 8T0 9 MEAN 16. 18.** 17. 18.* S. D. 1.2 1.1 1.0 0.8 N 10 10 10 10 _______________________________________ * = Significantly different from the control group at 0.05 using Dunnett's test ** = Significantly different from the control group at 0.01 using Dunnett's test 82 of 619 83 of 619 84of619 85 of 619 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE ANALYSIS GROUP: ______________ WHITE CELL (thous/ uL) MEAN S. D. N RED CELLS (mil/ uL) MEAN S. D. N HEMOGLOBIN (g/ dL) MEAN S. D. N HEMATOCRIT (%) MEAN S. D. N MCV (fL) MEAN S. D. N TABLE 9 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF HEMATOLOGY VALUES PAGE 1 WEEK 13 0 PPM _ MALE _____ 15 PPM 40 PPM 100 PPM 7.3 7.3 7.1 6.6 1.43 1.27 1.36 1.18 10 10 10 10 8.44 8.64 8.50 8.60 0.522 0.357 0.409 0.620 10 10 10 10 14.7 15.2 14.7 15.0 0.93 0.66 0.74 0.98 10 10 10 10 41.1 41.8 40.8 41.9 2.49 1.84 2.50 3.06 10 10 10 10 48.7 48.4 47.9 48.7 0.71 0.73 1.80 0.53 10 10 10 10 ______________________________________ g/ dL = GRAMS/ DECILITER, thous/ uL = THOUSANDS/ MICROLITER, mil/ uL = MILLIONS/ MICROLITER, uug = PICOGRAMS, fL = FEMTOLITERS ________________________________________

None significantly different from control group PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 9 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF HEMATOLOGY VALUES PAGE 2 WEEK 13 ANALYSIS GROUP:

MCH (uug) MEAN S. D. N MCHC (g/ dL) MEAN S. D. N PLATELET (thous/ uL) MEAN S. D. N 0 PPM 17.4 0.25 10 35.8 0.42 10 906. 192.7 10 ____ MALE _____ 15 PPM 40 PPM 100 PPM 17.6 17.3 17.4 0.23 0.51 0.31 10 10 10 36.3 36.0 35.8 0.34 0.43 0.47 10 10 10 900. 873. 929. 202.2 262.8 262.5 10 10 10 PROTIME (seconds)

3 MEAN S. D.

r?, N CA G APTT (seconds) MEAN S. D. N 13.9 13.7 13.9 13.6 1.10 0.78 1.04 0.46 9 8 9 7 24.0 24.7 23.2 23.5 4.29 3.19 4.43 2.85 9 8 9 7 _______________________________________

g/ dL = GRAMS/ DECILITER, thous/ uL = THOUSANDS/ MICROLITER, mil/ uL = MILLIONS/ MICROLITER, uug = PICOGRAMS, fL = FEMTOLITERS ________________________________________

None significantly different from control group PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 9 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF HEMATOLOGY VALUES PAGE 3 WEEK 13 ___ __  e F E M A L E _____ ANALYSIS GROUP: 0 PPM 15 PPM 40 PPM 100 PPM ________________________________________

WHITE CELL (thous/ uL) MEAN 5.3 5.6 5.2 4.1 S. D. 1.13 1.44 1.49 1.06 N 8 8 8 9 RED CELLS (mil/ uL) MEAN 7.65 7.69 7.71 7.87 S. D. 0.311 0.400 0.211 0.394 N 8 8 8 9 HEMOGLOBIN (g/ dL) MEAN S. D. N 14.5 14.2 14.5 14.7 0.59 1.06 0.29 0.72 8 8 8 9 HEMATOCRIT (%)

E MEAN 39.9 40.1 39.9 S. D. 1.67 2.21 1.05 41.3 2.11 0, N m 55 MCV (fL) MEAN 52.2 52.2 51.7 52.4 8 8 9 S. D. 0.56 0.78 0.7% 0.88 N 8 8 8 9 ______ ______________________________________

g/ dL = GRAMS/ DECILITER, thous/ uL = THOUSANDS/ MICROLITER, mil/ uL = MILLIONS/ MICROLITER, uug = PICOGRAMS, fL = FEMTOLITERS ________________________________________

None significantly different from control group 89 of 619 s i*` 0 co .ri m NO IO .! I 3 l 90of619 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 10 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF LEUKOCYTE DIFFERENTIAL COUNTS [% I PAGE 2 WEEK 13 w F E M A L E _____

ANALYSIS GROUP: 0 PPM 15 PPM ______________________________________ NEUTROPHIL MEAN 17 15 S. D. 3.3 3.5 N 8 8 LYMPHOCYTE MEAN 81 83 S. D. 4.2 4.9 N 8 8 MONOCYTE MEAN 2 2 S. D. 1.4 1.8 N 8 8 EOSINOPHIL a MEAN 0 0 r 2 S. D. N 0.4 8 0.0 8 z BASOPHIL MEAN 0 0 S. D. 0.0 0.0 N 8 8 None significantly different from control group _____ 40 PPM 100 PPM _________________

15 15 3.8 2.4 8 9 81 81 4.7 3.7 8 9 3 3 1.8 1.5 8 9 0 0 0.0 0.4 8 9 0 0 0.0 0.0 8 9 ____________

PCPSV4.04 01/ 18/ 2002 "22 l 4 . 0 92of619 I , PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 12 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF SERUM CHEMISTRY VALUES PAGE 1 WEEK 13 ___________________ __ MALE ___ ANALYSIS GROUP: 0 PPM 15 PPM 40 PPM 100 PPM ALBUMIN (g/ dL) MEAN 4.2 4.3 4.2 4.3 S. D. 0.26 0.25 0.27 0.32 N 10 10 10 10 TOTAL PROTEIN (g/ dL) MEAN S. D. N 7.3 7.4 7.2 7.3 0.60 0.48 0.51 0.65 10 10 10 10 GLOBULIN (g/ dL) MEAN S. D. N 3.1 3.1 3.0 3.0 0.38 0.26 0.29 0.34 10 10 10 10 A/ G RATIO MEAN 1.38 1.38 1.40 1.42 S. D. 0.123 0.077 0.089 0.074 N 10 10 10 10 TOTAL BILI (mg/ dL) MEAN S. D. N 0.2 0.2 0.1 0.2 0.11 0.10 0.07 0.11 10 10 10 10 _______________________________________ U/ L = INTERNATIONAL UNIT/ LITER, g/ dL = GRAMS/ DECILITER, mg/ dL = MILLIGRAMS/ DECILITER, mEq/ L = milliEquivalents/ Liter _____________________________________ None significantly different from control group 95 of 619 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 12 YO DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF SERUM CHEMISTRY VALUES PAGE 3 WEEK 13 ___ MALE __ ANALYSIS GROUP: 0 PPM 15 PPM 40 P PM 100 PPM ______________________________________

GLUTAMYLTRANSFER (U/ L) MEAN 0. 0. 0. 0. S. D. 0.0 0.0 0.0 0.0 N 10 10 10 10 GLUCOSE (mg/ dL) MEAN 152. 151. 154. 146. S. D. 16.1 20.8 18.3 18.2 N 10 10 10 10 CHOLESTEROL (mg/ dL) MEAN 53. 58. 55. 63.** S. D. 5.3 7.2 5.6 3.4 N 10 10 10 10 CALCIUM (mg/ dL) z MEAN 11.5 11.6 11.7 11.8 S. D. 0.51 0.54 0.40 0.70 on, N 10 10 10 10 a G CHLORIDE (mEq/ L) MEAN 101. 100. 100. 100. S. D. 1.4 1.1 1.5 1.7 N 10 10 10 10 ______________________________________

U/ L = INTERNATIONAL UNIT/ LITER, g/ dL = GRAMS/ DECILITER, mg/ dL = MILLIGRAMS/ DECILITER, mEq/ L = milliEquivaIents/ Liter _____________________________________ ** = Significantly different from the control group at 0.01 using Dunnett's test 97 of 619 TABLE 12 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 5 SPONSOR: HAP TASK FORCE SUMMARY OF SERUM CHEMISTRY VALUES WEEK 13 ANALYSIS GROUP: _____________

ALBUMIN (g/ dL) MEAN S. D. N TOTAL PROTEIN (g/ dL) MEAN S. D. N GLOBULIN (g/ dL) MEAN S. D. N A/ G RATIO MEAN S. D. N TOTAL BILI (mg/ dL) MEAN S. D. N e F E M A L E ____

0 PPM 15 PPM 40 PPM 100 PPM ___________________~

3.9 3.9 4.1 4.2 0.17 0.27 0.23 0.20 9 9 9 9 6.7 6.6 7.0 7.0 0.36 0.49 0.45 0.53 8 9 9 9 2.7 2.7 2.9 0.24 0.24 0.24 8 9 9 1.44 1.48 1.42 1.49 0.099 0.073 0.069 0.118 8 9 9 9 0.2 0.2 0.2 0.2 0.10 0.10 0.20 0.20 8 9 9 9 ________________________________________ U/ L = INTERNATIONAL UNIT/ LITER, g/ dL = GRAMS/ DECILITER, mg/ dL = MILLIGRAMS/ DECILITER, mEq/ L = milliEquivalents/ Liter None significantly different from control group PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE ANALYSIS GROUP: ____________ UREA NITROGEN (mg/ dL) MEAN S. V. N CREATININE (mg/ dL) MEAN S. D. N ALKALINEPHOS'TSE (U/ L) MEAN S. D. N ALANINE TRANSFER (U/ L) MEAN S. D. N ASPARTATTRANSFER (U/ L) MEAN S. D. N __ TABLE 12 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF SERUM CHEMISTRY VALUES PAGE 6 WEEK 13 _______________________________________  _ _ _ _ _ _  _

 w F E M A L E ____

0 PPM 15 PPM 40 PPM 100 PPM 16.2 17.0 17.5 16.9 2.77 0.72 1.41 1.67 8 9 9 9 0.2 0.2 0.2 0.2 0.05 0.07 0.07 0.08 8 9 9 9 65. 68. 70. 71. 12.4 6.2 7.5 7.5 9 9 9 9 57. 21.7 9 50. 55. 49. 6.8 11.9 8.5 9 9 9 138. 111. 93. 114. 83.1 45.7 17.8 69.5 9 9 9 9 U/ L = INTERNATIONAL UNIT/ LITER, g/ dL = GRAMS/ DECILITER, mg/ dL = MILLIGRAMS/ DECILITER, mEq/ L = milliEquivalents/ Liter ________________________________________ None significantly different from control group lOOof 101 of619 102of619 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 14 (WEEK 12 EVALUATION) 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE SUMMARY OF OPHTHALMIC EXAMINATION FINDINGS PAGE 1 _______________________________________~ NUMBER OF ANIMALS IN DOSE GROUP NUMBER OF ANIMALS EXAMINED EXAM 2( WEEK 12)

CORNEAL CRYSTALS  BILATERAL CORNEAL CRYSTALS  UNILATERAL KERATITIS  UNILATERAL CATARACT  UNILATERAL _________________

l 0 PPM 2 15 PPM 3 __ M A L E  ____ F E M A L E  GROUP: 1 2 3 4 1 2 3 4 _________ 10 10 10 10 10 10 10 10 10 10 10 10 10 9 10 10 10 10 10 10 9 9 10 10 0 0 0 0 2 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 0 0 0 _______________________________________ ____ 40 PPM 4 100 PPM POPSIv4.0 01/ 18/ 2002 104of619 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 16 (PRIMARY NEcRopsY) 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE GROSS NECROPSY OBSERVATIONS INCIDENCE SUMMARY PAGE 1 SCHEDULED NECROPSY ___________

GROUP: 1 2 NUMBER OF ANIMALS IN DOSE GROUP NUMBER OF ANIMALS EXAMINED WEEK M A L E   F E M A L E  3 4 1 2 3 4 _________________ _ ______ __ 10 9 0 0 1 0 0 0 0 0 1 10 10 0 0 0 0 0 0 0 0 0 EYES/ OPTIC N. EYE( S) OPACITY LIVER AREA( S), RAISED LUNGS AREA( S), WHITE LYMPH NODE, TR/ BR DISCOLORATION, DARK RED LYMPH NODE, MED % DISCOLORATION, DARK RED o\ OVARIES G AREA( S), WHITE PANCREAS PALE THYMUS DISCOLORATION, DARK RED PITUITARY SMALL ________________ l 0 PPM 2 NA = NOT APPLICABLE 10 10 10 10 10 10 13 10 10 10 10 10 10 0 0 0 0 0 0 0 1 1 1 0 1 1 0 0 0 1 1 0 0 NA NA NA 1 0 1 0 0 0 0 0 0 0 0 0 0 15 PPM 3 40 PPM 4 100 PPM TABLE 16 (PRIMARY NECROPSY) PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 2 SPONSOR: HAP TASK FORCE GROSS NECROPSY OBSERVATIONS INCIDENCE SUMMARY SCHEDULED NECROPSY __ M A L E  ___ F E M A L E  GROUP: 1 2 3 4 1 2 3 4 ________________________________________ NUMBER OF ANIMALS NUMBER OF ANIMALS SEMINAL VESICLES IN DOSE GROUP EXAMINED WEEK 10 10 0 0 2 0 0 0 0 10 10 10 10 10 10 10 9 10 10 10 10 1 0 0 NA NA NA NA 0 0 0 0 1 1 0 0 0 2 0 0 0 0 1 0 1 0 0 0 0 1 0 0 0 0 0 0 1 NA NA NA 6 8 6 0 9 0 6 0 0 0 0 0 0 0 10 13 SMALL SKELETAL MUSCLE DISCOLORATION,

SKIN MATTING, RED DARK RED MATTING, YELLOW SPLEEN ACCESSORY STOMACH CONTENTS, DARK RED LYMPH NODE, MAN DISCOLORATION, DARK RED UTERUS CONTENTS, CLEAR FLUID NA NO SIGNIFICANT CHANGES OBSERVED  ALL EXAMINED TISSUES 8 ___________________

PGRSIv4.02 11/ 28/ 2001 _____________________________________ l o PPM 2 15 PPM 3 40 PPM 4 NA = NOT APPLICABLE ____ 100 PPM 107 of 619 108of619 109of619 llOof619 TABLE 17 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 5 SPONSOR: HAP TASK FORCE ORGAN WEIGHTS (GRAMS)  SUMMARY OF MEANS WEEK 13 _ _ _ _ _ _ _ _ _ SPLEEN OVARIES THYMUS GROUP: 0 PPM _____ F E M A L E ___

15 PPM _ 40 PPM 100 PPM __________________________

MEAN 0.48 0.45 0.46 0.45 S. D. 0.030 0.050 0.022 0.027 N 10 9 10 10 MEAN 0.0957 0.0989 S. D. 0.00928 0.01719 N 10 9 MEAN 0.2163 0.1910 0.2042 0.2048 S. D. 0.02952 0.02205 0.01427 0.02387 N 10 9 10 10 ADRENAL GLANDS F F MEAN 0.0536 0.0589 F S. D. 0.00691 0.00529 0 N 10 9 2 G THYROIDS/ PARA MEAN 0.0150 0.0135 S. D. 0.00316 0.00187 N 10 9 ______________________________________ None significantly different from control group 0.0925 0.0977 0.01308 0.01334 10 10 0.0544 0.00479 10 0.0141 0.0142 0.00246 0.00349 10 10 __

POFBSTv4.1 11/ 28/ 2001 TABLE 18 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 1 SPONSOR: HAP TASK FORCE ORGAN WEIGHTS RELATIVE TO FINAL BODY WEIGHTS (GRAMS/ 100 GRAMS) WEEK 13 ________________________________________ _____ MALE __ GROUP: 0 PPM 15 PPM 40 PPM 100 PPM ________________________________________ FINAL BODY WT( G) MEAN S. D. N BRAIN LIVER KIDNEYS w L oy cn LUNGS ;; MEAN 0.636 0.629 0.634 0.609 S. D. 0.0347 0.0450 0.0466 0.0228 N 10 10 10 10 MEAN 2.791 2.809 2.707 2.791 S. D. 0.1170 0.1521 0.1315 0.1323 N 10 10 10 10 MEAN 0.682 0.696 S. D. 0.0303 0.0315 N 10 10 288. 302. 294. 305. 17.9 23.5 28.3 12.5 10 10 10 10 MEAN 0.416 0.413 S. D. 0.0231 0.0224 N 10 10 ________________________________________ __ None significantly different from control group __ 0.715 0.712 0.0362 0.0348 10 10 0.399 0.400 0.0318 0.0154 10 10 _______________________________________ PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 18 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE ORGAN WEIGHTS RELATIVE TO FINAL BODY WEIGHTS (GRAMS/ 100 GRAMS) PAGE 2 WEEK 13 ________________________________________ _____ MALE ____ GROUP: 0 PPM 15 PPM 40 PPM 100 PPM _______________________________________ ____

HEART MEAN 0.326 0.317 0.328 0.321 S. D. 0.0246 0.0130 0.0136 0.0182 N 10 10 10 9 SPLEEN MEAN S. D. N 0.198 0.203 0.198 0.205 0.0264 0.0123 0.0159 0.0159 10 10 10 10 EPIDIDYMIDES MEAN 0.339 0.326 0.327 0.332 S. D. 0.0253 0.0228 0.0187 0.0200 N 10 10 10 10 TESTES +

t; MEAN S. D. 0 N 1.055 1.036 1.060 1.029 0.0459 0.0770 0.0735 0.0364 10 10 10 10 THYMUS G MEAN 0.063 0.067 0.065 0.066 S. D. 0.0047 0.0060 0.0107 0.0074 N 10 10 10 10 ________________ None significantly different from control group 114 of 619

115of619 TABLE 18 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 5 SPONSOR: HAP TASK FORCE ORGAN WEIGHTS RELATIVE TO FINAL BODY WEIGHTS (GRAMS/ 100 GRAMS) WEEK 13 _____________________________________ ____ ____ F E M A L E ___ GROUP: 0 PPM 15 PPM 40 PPM 100 PPM _____________________________________

HEART MEAN 0.348 0.359 0.354 0.375* S. D. 0.0187 0.0217 0.0239 0.0204 N 10 9 10 10 SPLEEN MEAN 0.256 0.255 0.251 0.251 S. D. 0.0128 0.0277 0.0207 0.0110 N 10 9 10 10 OVARIES MEAN 0.051 0.056 0.051 0.055 S. D. 0.0058 0.0103 0.0088 0.0066 N 10 9 10 10 THYMUS + MEAN 0.115 0.108 0.112 0.115 z S. D. 0.0120 0.0147 0.0075 0.0131 0, N 10 9 10 10 cn ADRENAL GLANDS 5; MEAN 0.028 0.033** 0.030 0.032 S. D. 0.0031 0.0035 0.0030 0.0028 N 10 9 10 10 * = Significantly different from the control group at 0.05 using Dunnett's test ** = Significantly different from the control group at 0.01 using Dunnett's test 117of619 PROJECT NO.: WIL 417002 S!? ONSOR: HAP TASK FORCE TABLE 19 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE ORGAN WEIGHTS RELATIVE TO BRAIN WEIGHTS (GRAMS/ 100 GRAMS) PAGE 1 WEEK 13 __ MALE ___ GROUP: 0 PPM 15 PPM 40 PPM 100 PPM _____________________________________ BRAIN WT (GRAMS) MEAN 1.82 1.89 1.85 1.86 S. D. 0.051 0.073 0.099 0.054 N 10 10 10 10 LIVER MEAN 439.593 448.832 429.621 458.881 S. D. 19.3561 40.0560 40.3947 30.4098 N 10 10 10 10 KIDNEYS MEAN 107.459 111.060 113.389 117.063 S. D. 5.1934 7.0516 10.2130 7.2201 N 10 10 10 10 LUNGS MEAN S. D. N 65.419 66.000 63.224 65.720 2.9922 6.8888 6.2541 3.1061 10 10 10 10 HEART MEAN 51.290 50.603 51.986 52.517 S. D. 3.1086 3.6834 2.9534 3.0131 N 10 10 10 9 __________________~~~ None significantly different from control group 119of619 120of619 121 of 619 122of619 TABLE 20 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 1 SPONSOR: HAP TASK FORCE INDIVIDUAL SURVIVAL AND DISPOSITION ________________________________________ TYPE OF AGE IN DATE OF DAYS ON DEATH WEEKS A DEATH STUDY ___. ANIMAL SEX _________ 69302 M 69306 M 69308 M 69310 M 69312 M 69314 M 69317 M 69322 M 69337 M 69343 M GROUP _ _ _ _ _   0 PPM 0 PPM 0 PPM 0 PPM 0 PPM 0 PPM 0 PPM 0 PPM 0 PPM 0 PPM _____________________

SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA .___ 30 AUG 01 30 AUG 01 30 AUG 01 31 AUG 01 31 AUG 01 31 AUG 01 31 AUG 01 31 AUG 01 30 AUG 01 30 AUG 01 69305 M 69316 M 69320 M 69323 M 69326 M 69335 M 69336 M % 69338 69345 M M cn 69347 M G 69303 M 69309 M 69313 M 69315 M 69324 M 69328 M 69339 M 69344 M 15 PPM 15 PPM 15 PPM 15 PPM 15 PPM 15 PPM 15 PPM 15 PPM 15 PPM 15 PPM SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA _ _ _ _ _ _ _ _ _ _ 21 21 21 21 21 21 21 21 21 21 21 21 21 21 21 21 21 21 21 21 30 AUG 01 30 AUG 01 31 AUG 01 31 AUG 01 30 AUG 01 31 AUG 01 31 AUG 01 30 AUG 01 31 AUG 01 30 AUG 01 40 PPM 40 PPM 40 PPM 40 PPM 40 PPM 40 PPM 40 PPM 40 PPM SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA 21 21 21 21 21 21 21 21 30 AUG 01 31 AUG 01 30 AUG 01 30 AUG 01 31 AUG 01 31 AUG 01 30 AUG 01 31 AUG 01 91 91 91 92 92 92 92 92 91 91 91 91 92 92 91 92 92 91 92 91 91 92 91 91 92 92 91 92 A = CALCULATED TO THE NEAREST WHOLE WEEK USING THE MEAN AGE IN WEEKS AT INITIATION OF DOSING (8) TABLE 20 go DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE INDIVIDUAL SURVIVAL AND DISPOSITION PAGE 2 TYPE OF AGE IN DATE OF DAYS ON DEATH WEEKS A DEATH STUDY SCHEDULED EUTHANASIA 21 30 AUG 01 91 SCHEDULED EUTHANASIA 21 31 AUG 01 92 SCHEDULED EUTHANASIA 21 31 AUG 01 92 SCHEDULED EUTHANASIA 21 30 AUG 01 91 SCHEDULED EUTHANASIA 21 30 AUG 01 91 SCHEDULED EUTHANASIA 21 31 AUG 01 92 SCHEDULED EUTHANASIA 21 31 AUG 01 92 SCHEDULED EUTHANASIA 21 30 AUG 01 91 SCHEDULED EUTHANASIA 21 31 AUG 01 92 SCHEDULED EUTHANASIA 21 30 AUG 01 91 SCHEDULED EUTHANASIA 21 31 AUG 01 92 SCHEDULED EUTHANASIA 21 30 AUG 01 91 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE ANIMAL SEX GROUP ________________________

69348 M 40 PPM 69350 M 40 PPM 69304 M 100 PPM 69307 M 100 PPM 69318 M 100 PPM 69319 M 100 PPM 69327 M 100 PPM 69329 M 100 PPM 69332 M 100 PPM 69334 M 100 PPM 69342 M 100 PPM 69346 M 100 PPM _.

A = CALCULATED TO THE NEAREST WHOLE WEEK USING THE MEAN AGE IN WEEKS AT INITIATION OF DOSING (8) PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE ANIMAL SEX GROUP TABLE 20 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE INDIVIDUAL SURVIVAL AND DISPOSITION TYPE OF AGE IN DATE OF DAYS ON DEATH WEEKS A DEATH STUDY PAGE 3 69354 69366 69367 69378 69384 69398 69404 69416 69419 69432 69362 69376 69377 69382 69390 L 69393 WI 69396 0 69409 69413 69435 io 69359 69369 69370 69380 69388 69394 69426 69428 _______________________________________ A = CALCULATED TO THE NEAREST WHOLE WEEK USING THE MEAN AGE IN WEEKS AT INITIATION OF DOSING (8) 0 PPM SCHEDULED EUTHANASIA 0 PPM SCHEDULED EUTHANASIA 0 PPM SCHEDULED EUTHANASIA 0 PPM SCHEDULED EUTHANASIA 0 PPM SCHEDULED EUTHANASIA 0 PPM SCHEDULED EUTHANASIA 0 PPM SCHEDULED EUTHANASIA 0 PPM SCHEDULED EUTHANASIA 0 PPM SCHEDULED EUTHANASIA 0 PPM SCHEDULED EUTHANASIA 15 PPM SCHEDULED EUTHANASIA 15 PPM SCHEDULED EUTHANASIA 15 PPM EUTHANIZED IN EXTREMIS 15 PPM SCHEDULED EUTHANASIA 15 PPM SCHEDULED EUTHANASIA 15 PPM SCHEDULED EUTHANASIA 15 PPM SCHEDULED EUTHANASIA 15 PPM SCHEDULED EUTHANASIA 15 PPM SCHEDULED EUTHANASIA 15 PPM SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA 40 PPM 40 PPM 40 PPM 40 PPM 40 PPM 40 PPM 40 PPM 40 PPM 21 21 21 21 21 21 21 21 21 21 21 21 12 21 21 21 21 21 21 21 21 21 21 21 21 21 21 21 31 AUG 01 31 AUG 01 30 AUG 01 31 AUG 01 30 AUG 01 30 AUG 01 31 AUG 01 30 AUG 01 30 AUG 01 31 AUG 01 31 AUG 01 30 AUG 01 30 JUN 01 31 AUG 01 31 AUG 01 30 AUG 01 30 AUG 01 30 AUG 01 31 AUG 01 30 AUG 01 30 AUG 01 31 AUG 01 30 AUG 01 31 AUG 01 30 AUG 01 31 AUG 01 31 AUG 01 31 AUG 01 92 92 91 92 91 91 92 91 91 92 92 91 30 92 92 91 91 91 92 91 91 92 91 92 91 92 92 92 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 20 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE INDIVIDUAL SURVIVAL AND DISPOSITION PAGE 4 ANIMAL SEX GROUP  _____  _____. 69433 F 40 PPM 69436 F 40 PPM TYPE OF DEATH SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA 69375 F 100 PPM SCHEDULED EUTHANASIA 69379 F 100 PPM SCHEDULED EUTHANASIA 69383 F 100 PPM SCHEDULED EUTHANASIA 69385 F 100 PPM SCHEDULED EUTHANASIA 69392 F 100 PPM SCHEDULED EUTHANASIA 69402 F 100 PPM SCHEDULED EUTHANASIA 69408 F 100 PPM SCHEDULED EUTHANASIA 69415 F 100 PPM SCHEDULED EUTHANASIA 69427 F 100 PPM SCHEDULED EUTHANASIA 69431 F 100 PPM SCHEDULED EUTHANASIA AGE IN WEEKS A ____ 21 21 21 21 21 21 21 21 21 21 21 21 DATE OF DAYS ON DEATH STUDY ________ ____________________ _____ 30 AUG 01 91 30 AUG 01 91 30 AUG 01 91 30 AUG 01 91 31 AUG 01 92 31 AUG 01 92 31 AUG 01 92 31 AUG 01 92 31 AUG 01 92 30 AUG 01 91 30 AUG 01 91 30 AUG 01 91 ~ A = CALCULATED TO THE NEAREST WHOLE WEEK USING THE MEAN AGE IN WEEKS AT INITIATION OF DOSING (8) PDEADv4.02 r x 01/ 18/ 2002 TABLE 21 PROJECT NO.: WIL 417002 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE PAGE 1 SPONSOR: HAP TASK FORCE INDIVIDUAL BODY WEIGHTS [Gl MALE GROUP: 0 PPM WEEK 2 1 0 1 2 3 4 5 ______________________________________~ ANIMAL 69302 69306 69308 69310 69312 69314 69317 69322 69337 69343 98. 99. 110. 111. 221. 239. 221. 242.

107. 107. 105. 90. 108. 110. 123. 128. 136. 137. 133. 134. 134. 114. 130. 132. 161. 201. 165. 206. 178. 203. 171. 186. 175. 205. 169. 194. 179. 211. 153. 187. 166. 195. 172. 200. 207. 230. 208. 225. 233. 251. 208. 224. 234. 252. 212. 230. 209. 226. 218. 234. 252. 257. 241. 234. 264. 238. 266. 245. 236. 248. 258. 268. 256. 229. 268. 250. 277. 257. 251. 259.

MEAN 105. 130. 169. 199. 217. 235. 248. 257. S. D. 6.8 7.0 8.0 8.2 10.2 10.3 11.4 13.0 N 10 10 10 10 10 10 10 10 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE WEEK _____

ANIMAL 69305 69316 69320 69323 69326 69335 69336 69338 69345 107. 99. 114. 104. 106. 106. 110. 108. 103. 69347 100.

MEAN 106. S. D. 4.5 N 10 1        _ _

130. 126. 145. 131. 130. 123. 136. 135. 126. 123.

131. 170. 203. 227. 243. 257. 267. 6.8 8.6 8.7 11.5 15.7 15.9 16.3 10 10 10 10 10 10 10 90 DAY INHAL.

0 ._____

165. 191. 210. 225. 169. 205. 232. 254. 188. 216. 239. 255. 171. 206. 230. 254. 176. 212. 242. 268. 168. 207. 233. 220. 175. 203. 232. 244. 171. 201. 219. 234. 157. 194. 223. 245. 161. 190. 208. 227. TABLE 21 STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE INDIVIDUAL BODY WEIGHTS [Gl PAGE 2 MALE GROUP: 15 PPM 1 2 3 4 5 233. 269. 265. 271. 284. 255. 257. 243. 259. 238. 240. 278. 275. 276. 291. 276. 262. 254. 270. 244. PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE WEEK _____ ANIMAL 69303 69309 69313 69315 69324 69328 69339 69344 69348 69350 __

101. 124. 160. 175. 195. 213. 225. 236. 101. 129. 169. 210. 232. 251. 265. 274. 104. 129. 174. 207. 229. 252. 268. 272. 106. 124. 157. 179. 198. 212. 221. 225. 106. 128. 167. 193. 219. 243. 261. 277. 111. 133. 175. 207. 229. 254. 269. 283. 103. 126. 163. 185. 202. 213. 218. 232. 117. 145. 191. 218. 239. 256. 273. 284. 114. 139. 176. 205. 228. 242. 257. 269. 107. 131. 170. 196. 219. 236. 246. 255. MALE GROUP: 40 PPM 0 1 2 3 4 5 MEAN 107. 131. 170. 198. 219. 237. 250. 261. S. D. 5.4 6.7 9.7 14.3 15.5 18.0 21.4 22.2 N 10 10 10 10 10 10 10 10 TABLE 21 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE INDIVIDUAL BODY WEIGHTS [Gl PAGE 3 PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE TABLE 21 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE INDJVIDUAL BODY WEIGHTS [Gl WEEK _ _ _ _ _ _ _ _ _ _ _ . ANIMAL 69304 69307 69318 69319 69327 69329 69332 69334 69342 69346 PAGE 4 2 1 0 MALE GROUP: 100 PPM 1 2 3 4 5 _________________~~~

100. 129. 168 205 233 256. 268. 286. 105. 129. 170 204 229 248. 265. 280. 104. 128. 166 189 214 235. 253. 263. 95. 117. 156 185 212 235. 248. 265. 112. 139. 178 195 216 231. NA 247. 108. 131. 170 188 207 224. 233. 250. 112. 135. 172 194 218 234. 245. 256. 115. 140. 180 200 224 236. 251. 260. 115. 140. 175 209 237 255. 266. 278. 95. 120. 161 195 218 235. 253. 262.

MEAN 106. 131. 170. 196. 221. 239. 254. 265. S. D. 7.6 8.0 7.4 7.9 9.7 10.5 11.3 12.9 N 10 10 10 10 10 10 9 10 NA = NOT APPLICABLE PROJECT NO.: WIL 417002 SPONSOR: HAP TASK FORCE WEEK _ __.

ANIMAL 69302 69306 69308 69310 69312 69314 69317 69322 69337 69343 6 7 ____________

272. 286. 278. 289. 265. 272. 241. 255. 290. 305. 253. 269. 291. 303. 264. 274. 259. 269. 264. 276.

MEAN 268. 280. 285. 292. 286. 299. 308. 313. S. D. 15.6 15.8 15.3 16.2 19.4 29.0 22.2 17.6 N 10 10 10 10 10 10 10 10 TABLE 21 90 DAY INHAL. STUDY OF 1,1,2 TCE IN RATS WITH PK IN RATS & MICE INDIVIDUAL BODY WEIGHTS [G]

MALE GROUP: 10 PAGE 5 8 9 _______________ 0 PPM 11 12 13 ____________________________________