STUDY TITLE 

2-BROM0-3,3,3-TRIFLUOROPROPENE, E-1-BROM0-3,3,3-TRIFLUOROPROPENE 

AND 1-BROM0-2-TRIFLUOROMETHYL-3,3,3-TRIFLUOROPROPENE: 

AN ACUTE (4-HOUR) INHALATION TOXICITY STUDY IN THE RAT 

VIA NOSE-ONLY EXPOSURE 

DATA REQUIREMENTS 

OECD Part 403 

OPPTS 870.1300 

AUTHOR 

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

STUDY COMPLETION DATE 

14 January 2004 

PERFORMING LAB ORA TORY 

Huntingdon Life Sciences 

Mettlers Road, P.O. Box 2360 

East Millstone, New Jersey 08875-2360 

LABORATORY PROJECT NUMBER 

02-5449 

SPONSOR 

American Pacific Corporation 

3770 Howard Hughes Parkway 

Suite 300 

Las Vegas, Nevada 89109 

TOTAL PAGES 

172 
Huntingdon Life Sciences 02-5449 

GLP COMPLIANCE STATEMENT 

Page 2 

Final Report 

This study was conducted in compliance with the OECD Principles of Good Laboratory 

Practices ENV /MC/CHEM (98) 17 and the United States Environmental Protection 

Agency's Good Laboratory Practice Standards 40 CFR Part 792 (TSCA). 

Study Director: £~ 14-JM,o4-- ~· ~ Gary M. offman, B.A., D.A.B.T. 
Huntingdon Life Sciences 

SCIENTISTS 

02-5449 

SIGNATURE PAGE 

Page 3 

Final Report 

The following Scientists were responsible for the overall conduct of this study. 

Departmental supervisory personnel are listed on the personnel page of this report 

(Appendix D). 

~~ - He:i.V.M.,Ph.D. 

Pathologist 

SCIENTIFIC REVIEW 

The following Scientists have reviewed and approved this report. 

CarolS. Auletta, MBA, D.A.B.T., R.A.C. 

Senior Director, Safety Assessment 

Sylv~h.D., Diplomate A.C.V.P. 

Senior Vice President, Safety Assessment 

Date 

/23 ~- dL( 

Date 
Huntingdon Life Sciences 02-5449 Page4 

Final Report 

QUALITY ASSURANCE STATEMENT 

Listed b,elow are the dates that this study was inspected by the Quality Assurance Unit of 

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

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

as can be reasonably established. 

Type of 

Inspection 

GLP Protocol 

Review 

Exposure, 

Monitoring, 

Equipment& 

Training Records 

Terminal Necropsy 

& Training 

Records 

Final Report & 

Study Data 

Date(s) of 

Inspection 

07 Jun 02 

17 Sep 02 

01 Oct 02 

06 10 Jan 03 

Nancy L. Beck 

Senior Quality Assurance Auditor 

Reported to 

Study Director and Management 

07 Jun 02 

17 Sep 02 

04 Oct 02 

10 Jan 03 

141ct"'"i?C~ J_col{ 

Date 
/ \ .. 

Huntingdon Life Sciences 02-5449 

2-BROM0-3,3,3-TRIFLUOROPROPENE, 

E-1-BROM0-3,3,3-TRIFLUOROPROPENE 

Page 5 

Final Report 

ANI) 1-BROM0-2-TRIFLUOROMETHYL-3,3,3-TRIFLUOROPROPENE: 

AN ACUTE (4-HOUR) INHALATION TOXICITY STUDY IN THE RAT 

VIA NOSE-ONLY EXPOSURE 

SUMMARY 

This study was designed to assess the potential and comparative acute inhalation toxicity 

of 2-Bromo-3,3,3-Trifluoropropene, E-1-Bromo-3,3,3-Trifluoropropene and 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene when administered via nose-only 

exposure. The test substances were initially administered as a vapor to groups of 

Sprague-Dawley CD® rats (5/sex/group) for four hours at a target concentration of 

25,000 ppm. These exposures were performed on 17 September 2002. Based on the 

resulting mortality, additional exposures were required. 2-Bromo-3,3,3-Trifluoropropene 

was exposed at a targeted concentration of 5,000 ppm for four hours. 

E-1-Bromo-3,3,3-Trifluoropropene was exposed at a targeted concentration of 

50,000 ppm for four hours. These exposures were performed on 21 October 2002. The 

exposure levels were determined using a MIRAN® Ambient Air Analyzer. 

Observations for signs of toxicity were conducted on all animals at 15-minute intervals 

during the first hour of exposure and hourly for the remainder of the exposure. Detailed 

physical observations were performed on all animals just prior to exposure, immediately 

upon removal from the chamber, hourly for two hours post-exposure and once daily 

thereafter. Body weight measurements were obtained just prior to exposure and weekly 

thereafter. After a 14-day post-exposure observation period, survivors were sacrificed. 

Complete macroscopic postmortem examinations were performed on all animals at the 

time of death or sacrifice. Selected organs were weighed for all terminally sacrificed 

animals and the lungs were examined microscopically for all test animals. 

The results of the exposures and subsequent mortality are summarized as follows: 

Test Substance Analytical Mortality Mortality Mortality 

Concentration Males Females Total 

(ppm) 

2-Bromo-3,3,3- 26,580 515 515 10/10 

Trifluoropropene 

2-Bromo-3,3,3- 5,173 0/5 0/5 0/10 

Trifluoropropene 

E-1-Bromo-3,3,3- 45,850 2/5 0/5 2/10 

Tri:fluoropropene 

E-1-Bromo-3,3,3- 25,650 015 0/5 0/10 

Trifluoropropene 

1-Bromo-2- 24,800 5/5 515 10/10 

Tri:fluoromethyl-3,3,3-

Trifluoropropene 
Huntingdon Life Sciences 02-5449 

SUMMARY 

Page 6 

Draft Final Report 

All of t~e animals exposed to 2-Bromo-3,3,3-Trifluoropropene at 26,580 ppm were found 

dead or were euthanized in poor condition on the day after the exposure. All of the 

animals exposed to 1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene at 24,800 ppm 

were found dead within 30 minutes of the start of the exposure. Both of the male animals 

exposed to E-1-Bromo-3,3,3-Trifluoropropene at 45,850 ppm that were found dead died 

on the day of the exposure. All other animals survived to termination. 

Based on the above results, the LC50 values were calculated to be: 

Test Substance Animals LCso 95% Confidence Limits 

(ppm) (ppm) 

2-Bromo-3,3 ,3- Combined 11726 9054 to 15186 

Trifluoropropene Males 11726 7584 to 18129 

Females 11726 7584 to 18129 

E-1-Bromo-3,3,3- Combined 70955 51449 to 97857 

Trifluoropropene Males 50950 35358 to 73418 

Females >45850 Not Calculable 

1-Bromo-2- Combined <24,800 Not Calculable 

Trifluoromethyl-3,3,3- Males <24,800 Not Calculable 

Trifluoropropene Females <24,800 Not Calculable 

All surviving animals in all test groups gained weight (or did not lose weight) during both 

weeks of observations. 

2-Bromo-3,3,3-Trifluoropropene: The only sign of toxicity noted during the exposure 

period was labored breathing or gasping during the last hour of each exposure. Clinical 

signs of toxicity noted immediately following the exposures of 

2-Bromo-3,3,3-Trifluoropropene included clear or red nasal discharge, excessive 

salivation, labored breathing and moist rales. The severity and frequency of the signs was 

increased after the higher level exposure of 26,580 ppm compared to the lower level 

exposure of 5,173 ppm. Gasping and decreased activity were also noted but only after the 

higher exposure level. There were no apparent effects of exposure on the organs that were 

weighed. Macroscopically, one or more rats from both exposure levels had red 

discolorations of the lungs. Fluid was present in the lungs of one male from the higher 

level exposure of 26,580 ppm. Test substance-related findings were microscopically 

present in lungs of rats in the higher level exposure of 26,580 ppm. All of the rats, in this 

group only, had bronchiolar lesions consisting of desquamated epithelium, 

bronchiolar/peribronchiolar acute/subacute inflammation and/or intraluminal debris; the 

severity of the findings ranged from slight to moderate. Alveolar/intraalveolar 

macrophages (minimal to moderate) normally present in the lungs, were most numerous 

in several of these rats. 
(·· 

Huntingdon Life Sciences 02-5449 

SUMMARY 

Page 7 

Draft Final Report 

E-1-Bromo-3,3,3-Trifluoropropene: The only sign of toxicity noted during the 

exposure period was tail quivering and/or tremors during each exposure. Clinical signs of 

toxicity noted immediately following the exposures of E-1-Bromo-3 ,3 ,3-Trifluoropropene 

included red nasal discharge. The severity and frequency of the signs was increased after 

the higher level exposure of 45,850 ppm compared to the lower level exposure of 

25,650 ppm. Excessive salivation, labored breathing and moist rales were also noted but 

only after the higher exposure level. A few signs of toxicity similar to those seen 

immediately after the exposures were noted during the first week of the post-exposure 

observation period. A complete recovery in surviving animals was seen during the 

remainder of the post-exposure observation period. There were no apparent effects of 

exposure on the organs that were weighed. Macroscopically, one or more rats from the 

lower level exposure of 25,650 ppm had red discolorations of the lungs. Fluid was present 

in the lungs of two males from the higher level exposure of 45,850 ppm. 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene: All animals were found dead 

within 30 minutes of the exposure initiation. Macroscopically, one or more rats had red 

discoloration of the lungs. 

In conclusion, the 4-hour acute inhalation toxicity in rats of 

2-Bromo-3 ,3,3-Trifluoropropene, E-1-Bromo-3 ,3 ,3-Trifluoropropene and 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene was compared. On the basis of toxic 

responses and subsequent mortality, E-1-Bromo-3,3,3-Trifluoropropene was considered 

the least toxic with an LC50 of ~70,955 ppm, 2-Bromo-3,3,3-Trifluoropropene was 

considered moderately toxic with an LC50 of ~ 11,726 ppm and 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene was considered the most toxic with 

an LCso substantially less than the only tested level of 24,800 ppm. 
Huntingdon Life Sciences 02-5449 

TABLE OF CONTENTS 

Page 8 

Final Report 

COVERPAGE ..................................................................................................................... 1 

GLP COMPLIANCE STATEMENT .................................................................................. 2 

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

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

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

TABLE OF CONTENTS ..................................................................................................... 8 

1. INTRODUCTION ................................................................................... ............. 10 

2. MATERIALS AND METHODS 

2.1. Study Management .................................................................................... 10 

2.2. Study Dates ................................................................................................ 10 

2.3. Experimental Outline ................................................................................. 11 

2.4. Justifications .............................................................................................. 12 

2.5. Test Substances .......................................................................................... 12 

2.6. Test Animals .............................................................................................. 14 

2.7. Animal Assignment ................................................................................... 15 

2.8. Animal Identification ................................................................................. 15 

2.9. Veterinary Care .......................................................................................... 15 

2.10. Husbandry During Non-Exposure Periods ................................................. 16 

2.11. Husbandry During Exposure ...................................................................... 17 

2.12. Test Substance Administration and Chamber Operation ........................... 18 

2.13. Exposure Chamber Sampling .................................................................... 22 

2.14. Experimental Evaluations ......................................................................... .23 

2.15. Postmortem ................................................................................................ 24 

2.16. Statistical Analysis ..................................................................................... 26 

2.17. Data Storage ............................................................................................... 26 

2.18. Regulatory References ............................................................................... 26 

2.19. Protocol Deviations .................................................................................... 27 

3. RESULTS AND DISCUSSION 

3.1. Chamber Monitoring .................................................................................. 28 

3.2. Mortality ..................................................................................................... 28 

3.3. Body Weights ............................................................................................. 29 

3.4. Physical Observations ................................................................................ 29 

3.5. Pathology ................................................................................................... 31 

4. CONCLUSION .................................................................................................... 32 
Huntingdon Life Sciences 

FIGU~S 

02-5449 

TABLE OF CONTENTS 

Page 9 

Final Report 

1. Diagram of Generation and Sampling System (Groups 1- 3) ..................................... 33 

2. Diagram of Generation and Sampling System (Group 4) ............................................ 34 

3. Diagram of Generation and Sampling System (Group 5) ............................................ 35 

TABLES 

1. Chamber Concentrations, Particle Size Determinations and Chamber Environment. .36 

2. Body Weights and Body Weight Changes from Day 0 ............................................... .41 

3. Summary of Day 0 Observations ................................................................................ .43 

4. Su:mn1ary of 14-Day Post-Exposure Observations ..................................................... .47 

5. Individual Observations ............................................................................................... 51 

6. Animal Termination History ........................................................................................ 61 

7. Mean Organ Weights ................................................................................................... 62 

8. Individual Organ Weights ............................................................................................ 69 

9. Incidence Summary Report for Gross Necropsy Observations .................................... 76 

10. Lesion Incidence Summary with Expanded Severity Levels ....................................... 80 

11. Individual Animal Gross and Microscopic Observations ............................................ 89 

APPENDICES 

A. Equipment List ........................................................................................................... 140 

B. MIRAN® Calibration ................................................................................................. 141 

C. Protocol and Protocol Amendments .......................................................................... 152 

D. Testing Facility Personnel. ......................................................................................... 171 

E. Report Amendments .................................................................................................. 172 
r 

I 

' 

Huntingdon Life Sciences 02-5449 Page 10 

Final Report 

1. INTRODUCTION 

2. 

The purpose of this study was to assess the potential and comparative acute 

inhalation toxicity of 2-Bromo-3,3,3-Trifluoropropene, E-1-Bromo-3,3,3-

Trifluoropropene and 1-Bromo-2-Trifluoromethyl-3 ,3 ,3-Trifluoropropene when 

administered for 4 hours via nose-only exposure. 

MATERIALS AND METHODS 

2.1. STUDY MANAGEMENT 

2.1.1. SPONSOR 

American Pacific Corporation 

3770 Howard Hughes Parkway 

Suite 300 

Las Vegas, Nevada 89109 

2.1.2. SPONSOR REPRESENTATIVE 

Bradford Colton 

2.1.3. TESTING FACILITY 

Huntingdon Life Sciences 

P.O. Box 2360 

Mettlers Road 

East Millstone, New Jersey 08875-2360 

2.1.4. STUDY DIRECTOR 

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

2.2. STUDY DATES 

2.2.1. STUDY INITIATION 

15 August 2002 (Date Study Director signed the Protocol) 
(: 

Huntingdon Life Sciences 02-5449 

2.2.2. DATES OF ANIMAL RECEIPT 

Page 11 

Final Report 

10 September 2002: Groups 1- 3 (Experimental Start Date) 

15 October 2002: Groups 4- 5 

2.2.3. DATES OF EXPOSURE 

17 September 2002: Groups 1 - 3 

21 October 2002: Groups 4- 5 

2.2.4. TERMINAL SACRIFICES 

01 October 2002: Groups 1- 3 

04 November 2002: Groups 4-5 

2.2.5. EXPERIMENTAL TERMINATION 

14 January 2004 (Date Final Report is signed by the Pathologist) 

2.2.6. STUDY COMPLETION 

\. •·· 14 January 2004 (Date Final Report is signed by the Study 

Director) 

2.3. EXPERIMENTAL OUTLINE 

Group Test Substance Target Duration of 

Concentration Exposure 

(ppm) (Hours) 

1 2-Bromo-3,3 ,3-Trifluoropropene 25,000 4 

2 E-1-Bromo-3,3,3-Trifluoropropene 25,000 4 

3 1-Bromo-2-Trifluoromethyl-3,3 ,3-Trifluoropropene 25,000 4 

4 2-Bromo-3,3,3-Trifluoropropene 5,000 4 

5 E-1-Bromo-3,3 ,3-Trifluoropropene 50,000 4 

Each group consisted of 5 male and 5 female Sprague-Dawley rats. 

All survivors were held for a 14-day post-exposure observation period. At 

the end of the observation period, a macroscopic and microscopic necropsy 

examination was performed on all animals. 
Huntingdon Life Sciences 02-5449 

2.4. JUSTIFICATIONS 

2.4.1. ROUTE, DURATION AND FREQUENCY 

Page 12 

Final Report 

The inhalation route is one of the potential routes of human 

exposure to these test substances and is the route· specified in the 

referenced guidelines. The duration of the study and frequency of 

exposures are as recommended in the referenced guidelines. 

2.4.2. TEST ANIMAL SELECTION 

The rat is a rodent animal model commonly utilized in toxicity 

testing as per the regulatory requirements. In addition, a historical 

database is available for comparative evaluation. 

2.4.3. NUMBER OF ANIMALS 

The number of animals in the study was considered to be the 

minimum necessary for scientific and statistical reasons in order to 

evaluate the data with sufficient confidence levels. 

2.4.4. EXPOSURE LEVEL SELECTION 

The exposure levels were specified by the Sponsor based on prior 

testing of these or similar chemicals. 

2.5. TEST SUBSTANCES 

2-Bromo-3 ,3 ,3-Trifluoropropene 

E-1-Bromo-3,3,3-Trifluoropropene 

1-Bromo-2-Trifluoromethyl-3,3 ,3-Trifluoropropene 

2.5.1. TEST SUBSTANCE CATEGORY 

Industrial Gases 

2.5.2. CAS NUMBER 

2-Bromo-3,3,3-Trifluoropropene: 1514-82-5 

E-1-Bromo-3,3,3-Trifluoropropene: 149597-47-7 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene: N/ A 
( 

Huntingdon Life Sciences 02-5449 

2.5.3. SUPPLIER 

American Pacific Corp. - Halotron Division 

10622 West, 6400 North 

Cedar City, Utah 84720 

2.5.4. LOT NUMBERS 

Page 13 

Final Report 

2-Bromo-3,3,3-Trifluoropropene: KGA154-2B 

E-1-Bromo-3,3,3-Trifluoropropene: KGA154-1A (Container#!) 

59-57 (Container #2) 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene: 98-27 

2.5.5. PURITY 

2-Bromo-3,3,3-Trifluoropropene: ~99.5% 

E-1-Bromo-3,3,3-Trifluoropropene: 98.3% (Container #l)a 

98% (Container #2)b 

1-Bromo-2-Trifluoromethyl-3,3 ,3-Trifluoropropene: ~ 99% 

2.5.6. DESCRIPTION 

Colorless volatile liquid 

2.5.7. DATES RECEIVED 

22 August 2002 (Container #1 of the 3 test substances) 

14 October 2002 (Container #2 of E-1-Bromo-3,3,3-

Trifluoropropene) 

2.5.8. EXPIRATION DATE 

Not available (stable, per MSDS) 

2.5.9. ANALYSIS 

Documentation of the identity, strength, purity, compos1t10n, 

stability, and method of synthesis, fabrication, and/or derivation of 

the test substances and the maintenance of these records were the 

responsibility of the Sponsor. 

"Other impurities in this lot are: 0.5% Z-1-bromo-3,3,3-trifluoropropene and 1.0% 2-bromo3 ,3 ,3-trifluoropropene. 

bOther impurities in this lot are: 0.4% Z-1-bromo-3,3,3-trifluoropropene and 0.8% 2-bromo3,3,3-trifluoropropene. 
( 

Huntingdon Life Sciences 02-5449 Page 14 

Final Report 

2.6. 

2.5.10. STORAGE 

Room temperature 

2.5.11. ARCHIVAL SAMPLE 

An archival sample of each test substance was not taken. 

2.5.12. DISPOSITION 

The unused portion of the test substances and any empty test 

substance containers were returned to the Sponsor on 

13 November 2002. 

TEST ANIMALS 

2.6.1. SPECIES 

Albino Rats (Outbred) V AF/Plus® 

Sprague-Dawley 

Crl:CD® (SD) IGS BR 

2.6.2. SUPPLIER 

Charles River Laboratories 

Kingston, New York 12484 

2.6.3. NUMBER OF ANIMALS 

Received: 

60 total (30 males, 30 females) 

Placed on test: 

50 total (25 males, 25 females) 

Females were nulliparous and non-pregnant. 

2.6.4. AGE AT RECEIPT 

Approximately 7 weeks 

2.6.5. AGE AT EXPOSURE 

Approximately 8 weeks 
Huntingdon Life Sciences 02-5449 

2.6.6. WEIGHT AT EXPOSURE (GRAMS) 

Male: 

Female: 

Mean 

256 

205 

Range 

227-280 

181-225 

Page 15 

Final Report 

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

the mean weight for each sex. 

2.6.7. ACCLIMATION PERIOD 

Animals were acclimated for one week. All animals were checked 

for viability twice daily. Prior to assignment to study, all animals 

were examined to ascertain suitability for study. 

2.7. ANIMAL ASSIGNMENT 

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

Animals were arbitrarily placed in cages upon receipt and were placed on 

study and assigned to groups as available at time of study initiation. 

Animals considered unsuitable for the study on the basis of pretest 

physical examinations and outlying body weight data were excluded. 

Disposition of all animals not utilized in the study were maintained on file 

at the Testing Facility. 

2.8. ANIMAL IDENTIFICATION 

Each rat was assigned a temporary identification number upon receipt. 

After selection for study, each rat was ear-tagged with a number assigned 

by the Testing Facility. The assigned animal number plus the study 

number comprised the unique animal number for each animal. In addition, 

each cage was provided with a cage card, which was color-coded for 

exposure level identification and contained study number and animal 

number information. 

2.9. VETERINARY CARE 

Animals were monitored by the technical staff for any conditions requiring 

possible veterinary care and treated as necessary. 
Huntingdon Life Sciences 02-5449 

2.10. HUSBANDRY DURING NON-EXPOSURE PERIODS 

Page 16 

Final Report 

2.10.1. FACILITIES MANAGEMENT/ANIMAL HUSBANDRY 

Currently acceptable practices of good animal husbandry were 

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

National Academy Press, 1996. Huntingdon Life Sciences, East 

Millstone, New Jersey is fully accredited by the Association for 

Assessment and Accreditation of Laboratory Animal Care 

International (AAALAC). 

2.10.2. HOUSING 

Rats were group-housed (2-6/cage) during acclimation, then 

individually in suspended stainless steel wire mesh cages during 

the study. 

2.10.3. FEED 

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

International, St. Louis, Missouri) was available without 

restriction. Fresh feed was presented as required. 

2.10.4. ANALYSIS OF FEED 

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

the manufacturer. Results are maintained on file by the 

manufacturer. There were no known contaminants in the feed 

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

2.10.5. WATER 

Water was available without restriction via an automated watering 

system (Elizabethtown Water Company). 

2.10.6. WATERANALYSIS 

Water analyses are conducted by Elizabethtown Water Company, 

Westfield, New Jersey (Raritan-East Millstone Plant) to ensure that 

water meets standards specified under the EPA Federal Safe 

Drinking Water Act Regulations (40 CFR Part 141). In addition, 

water samples are collected biannually from representative rooms 

in the Testing Facility; chemical and microbiological water 
. _j 

Huntingdon Life Sciences 02-5449 Page 17 

Final Report 

analyses are conducted on these samples by a subcontract 

laboratory. Results of all water analyses are maintained on file at 

the Testing Facility. There were no known contaminants in the 

water which were expected to interfere with the results of this 

study. 

2.10.7. ENVIRONMENTAL CONDITIONS 

Light/Dark Cycle 

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

was provided. 

Temperature 

Temperature was monitored in accordance with the Testing 

Facility SOPs and maintained within the specified range to the 

maximum extent possible. 

Desired Range: 

Actual Range: 

Daily Average Range: 

Relative Humidity 

18 to 26°C 

21.0 to 23.5°C 

21.0 to 22.6°C 

Relative humidity was monitored in accordance with the Testing 

Facility SOPs and maintained within the specified range to the 

maximum extent possible. Excursions outside the specified range 

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

Desired Range: 

Actual Range: 

Daily Average Range 

30 to 70% 

19.06 to 72.49% 

25.62 to 57.27% 

2.11. HUSBANDRY DURING EXPOSURE 

2.11.1. HOUSING 

Animals were individually housed in polycarbonate tubes attached 

to a flow-past nose-only exposure chamber. 
Huntingdon Life Sciences 02-5449 

2.11. HUSBANDRY DURING EXPOSURE 

2.11.1. HOUSING 

Page 18 

Final Report 

Animals were individually housed in polycarbonate tubes attached 

to a flow-past nose-only exposure chamber. 

2.11.2. FEED 

None was provided during exposure. 

2.11.3. WATER 

None was provided during exposure. 

2.11.4. ENVIRONMENTAL CONDITIONS 

Chamber temperature and relative humidity were recorded at the 

initiation of exposure equilibrium period, at initiation of 4-hour 

exposure and every half-hour during exposure and maintained, to 

the maximum extent possible, within the ranges presented below. 

Excursions outside the specified ranges were not considered to 

have affected the integrity of the study. 

Temperature 

Desired Range: 

Actual Range: 

Relative Humidity 

Desired Range: 

Actual Range: 

20 to 24°C 

15 to 20°C 

30 to 70% 

34 to 58% 

2.12. TEST SUBSTANCE ADMINISTRATION AND CHAMBER 

OPERATION 

2.12.1. ROUTE OF ADMINISTRATION 

Inhalation, via nose-only exposure. 

2.12.2. FREQUENCY AND DURATION OF ADMINISTRATION 

The test substances were each administered for 4 hours for 

1 exposure. 
Huntingdon Life Sciences 02-5449 

2.12.3. PRE-STUDY TRIALS 

Page 19 

Final Report 

Trials were performed to evaluate the optimal set of conditions and 

equipment to generate stable atmospheres at the targeted exposure 

levels. 

2.12.4. CHAMBER OPERATION 

The exposure chambers were operated dynamically under 

equilibrium pressure. The total chamber airflow rate and 99% 

equilibrium time (T99) for each group are summarized below: 

Group Airflow Rate T99 

(Lpm) (min) 

1 10.0 3 

2 10.0 3 

3 10.0 3 

4 11.1 2 

5 6.5 4 

This chamber size and airflow rate was considered adequate to 

maintain the oxygen above 19%. The chamber was exhausted 

through a system consisting of a coarse filter, a HEPA filter and an 

activated charcoal bed. 

All animals remained in the chamber for a minimum of 30 minutes 

(Group 3 remained in the chamber for only 3 minutes) at the end of 

the exposure. During this time the chamber was operated at the 

same flow rate as used during the exposure using clean air only. 

Recordings of airflow rate and static pressure were made T99 prior 

to initiation, at initiation and every half-hour during exposure. 

Refer to Figures 1, 2 and 3 and Appendix A for equipment details. 

2.12.5. EXPOSURE PROCEDURE 

Groups 1-3 

For each exposure, the test substance was placed in a glass bubbler 

with a fritted bottom. 
Huntingdon Life Sciences 02-5449 Page 20 

Final Report 

House-line air was delivered from a regulator and a backpressure 

gauge, via 1

4" tubing, to a stainless steel "T" tube, which split the 

airflow into the generation and dilution systems. 

The generation air was directed via W' tubing from the stainless 

steel "T' tube, through a flowmeter, regulated by a metering valve. 

The outlet of the flowmeter was attached to the inlet of the glass 

bubbler via %" tubing. The test substance laden air stream was 

directed from the glass bubbler to one side of a second stainless 

steel "T" tube via W' Teflon tubing . 

The dilution air was directed, via W' tubing, from the first stainless 

steel "T" tube, to a flowmeter regulated by a metering valve. The 

air was then directed to the opposite side of the second stainless 

steel "T" and then into the inlet of the MIR.AN®. From the outlet 

of the MIRAN®, the test substance laden airflow was directed into 

the inlet of the flow-past nose-only exposure chamber. 

Refer to Figure 1 and Appendix A for equipment details. 

The generation and dilution flowrates are summarized in the 

following table. 

Group 4 

The test substance was placed in a glass bubbler with a fritted 

bottom and covered in foil. 

House-line nitrogen was delivered at a rate of 0.1 Lpm from a 

regulator and a backpressure gauge, via %" tubing, and connected 

to the inlet of the generation flowmeter, which was regulated by a 

metering valve. The outlet of the generation flowmeter was 

attached to the inlet of the glass bubbler via %" tubing. The test 

substance laden air stream was then directed, via %" Teflon tubing, 

from the outlet of the glass bubbler to one side of a stainless steel 

"T." 

House-line air was delivered at a rate of 11.0 Lpm from a regulator 

and a backpressure gauge, via %" tubing, to the inlet of the dilution 

flowmeter, which was regulated by a metering valve. The air was 

directed from the flowmeter to the opposite side of the stainless 
( 

Huntingdon Life Sciences 02-5449 Page 21 

Final Report 

steel "T" outlined above. From the "T," the test substance laden 

air stream was directed into the inlet of the MIRAN®. From the 

outlet of the MIRAN®, the test substance laden airflow was 

directed into the inlet of the flow-past nose-only exposure 

chamber. 

Refer to Figure 2 and Appendix A for equipment details. 

The generation and dilution flowrates are summarized in the 

following table. 

Group 5 

The test substance was placed in a glass bubbler with a fritted 

bottom and covered in foil. 

House-line air was delivered from a regulator and a backpressure 

gauge, via %" tubing, to a stainless steel "T" tube, which split the 

airflow into the generation and dilution systems. 

For the generation system, a flowmeter was connected to a 

stainless steel "T" via %" tubing. A metering valve was attached to 

the flowmeter's inlet to maintain the flowrate at 0.5 Lpm. 

A second generation flow used house-line nitrogen delivered from 

a regulator and a backpressure gauge, via %" tubing, and connected 

to the opposite side of the stainless steel "T" described above. A 

metering valve was attached to the inlet of the flowmeter to 

maintain the flowrate at 0.5 Lpm. 

The outlet of the house-line air/nitrogen ''T" was attached to the 

inlet of the glass bubbler via %" tubing. The test substance laden 

air stream was then directed from the bubbler outlet to one side of 

a second stainless steel "T" via %" Teflon tubing. 

House-line air was delivered from a regulator and a backpressure 

gauge, via W' tubing, and connected to the inlet of the dilution 

flowmeter, which was regulated by a metering valve. The airflow 

of 5.5 Lpm was directed from the flowmeter to the opposite side of 

the second stainless steel "T." From the "T," the test substance 

laden air stream was directed into the inlet of the MIRAN®. From 

the outlet of the MIRAN®, the test substance laden airflow was 
Huntingdon Life Sciences 02-5449 Page 22 

Final Report 

directed into the inlet of the flow-past nose-only exposure 

chamber. 

Refer to Figure 3 and Appendix A for equipment details. 

The generation and dilution flowrates are summarized in the table 

below: 

Group Generation Flow Dilution Flow 

(Lpm) (Lpm) 

Total Air Nz 

1 0.6 0.6 0.0 9.4 

2 0.75 0.75 0.0 9.25 

3 1.4 1.4 0.0 8.6 

4 0.10 0.0 0.10 11.0 

5 1.0 0.5 0.5 5.5 

The exposure chambers were enclosed m a 10 m3 glass and 

stainless steel chamber. 

2.13. EXPOSURE CHAMBER SAMPLING 

2.13.1. CHAMBER SAMPLING 

Determination of the exposure levels were made using a MIRAN® 

Ambient Air analyzer equipped with a strip chart recorder and 

digital multimeter. The test atmospheres were continuously 

generated through the MIRAN® and measurements were recorded 

4 times during the exposures. The exposure levels were 

determined by comparison of the measured absorbance to a 

calibrated response curve constructed using the same instrument 

settings. 

Refer to Appendix A for equipment details. 

2.13.2. PARTICLE SIZE DISTRIBUTION ANALYSIS 

Particle size samples were drawn at 4 intervals during each 

exposure for the chambers and room air using a TSI Aerodynamic 

Particle Sizer. The samples were drawn for 20 seconds at a rate of 

5.0 Lpm. The mass median aerodynamic diameter, geometric 
Huntingdon Life Sciences 02-5449 Page 23 

Final Report 

standard deviation and total mass concentration were calculated. A 

computer was used to program the system to the appropriate 

settings prior to sampling. The particle size distributions were 

calculated by the computer and printed out. 

Refer to Appendix A for equipment details. 

2.13.3. NOMINAL CONCENTRATION 

The nominal concentrations (ppm) were determined by weighing 

the generation apparatus containing the test substance before and 

after the exposure and calculating as follows: 

Cone (ppm)= 

amount consumed (g) x 1000 (mg/g) x 1000 (Ug/mg) x 22.4 (uUumole) x 295°K 

exposure duration (min) x airflow (Lpm) x MW (J.lg/J.lmole) x 273°K 

The molecular weight (MW) of each test substance is as follows: 

2-Bromo-3,3,3-Trifluoropropene = 174.95 

E-1-Bromo-3,3,3-Trifluoropropene = 174.95 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene = 242.95 

2.14. EXPERIMENTAL EVALUATIONS 

2.14.1. VIABILITY CHECKS 

Animals were observed in their cages for mortality and general 

condition twice daily (once in the morning and once in the 

afternoon). 

2.14.2. PHYSICAL EXAMINATIONS 

Examinations included but were not limited to, evaluation of skin 

and fur, eyes and mucous membranes, respiratory and circulatory 

effects, autonomic effects such as salivation, central nervous 

system effects, including tremors and convulsions, changes in the 

level of motor activity, gait and posture, reactivity to handling or 

sensory stimuli, grip strength, and stereotypies or bizarre behavior 

(e.g., self-mutilation, walking backwards). 
Huntingdon Life Sciences 02-5449 

Day 0 (Day of Exposure) 

Page 24 

Final Report 

Animals were observed individually prior to exposure, as a group 

at fifteen-minute intervals during the first hour of exposure, and 

hourly for the remainder of the exposure. They were observed 

individually upon removal from the chamber and hourly for 

two hours post-exposure. 

Days 1-14 

Detailed general observations were recorded once daily. 

2.14.3. BODY WEIGHT 

Animals were removed from their cages and weighed on Day 0 

(just prior to exposure), Day 7 and Day 14 (just prior to sacrifice). 

2.15. POSTMORTEM 

2.15.1. NECROPSY INFORMATION 

All survivors (up to 5/sex/group) were euthanized by 

exsanguination following carbon dioxide inhalation at termination 

on Test Day 14. 

2.15.2. MACROSCOPIC EXAMINATIONS 

Complete macroscopic postmortem examinations were performed 

on all animals. The macroscopic postmortem examination 

included examination of the external surface and all orifices; the 

organs and tissues of the cranial, thoracic, abdominal and pelvic 

cavities and neck; and the remainder of the carcass for the presence 

of macroscopic morphologic abnormalities. 

2.15.3. ORGAN WEIGHTS 

Organs indicated in Table I (page 25) were weighed for all animals 

at the scheduled sacrifice interval. Prior to weighing, the organs 

were carefully dissected and properly trimmed to remove adipose 

and other contiguous tissues in a uniform manner. Organs were 

weighed as soon as possible after dissection in order to avoid 

drying. Paired organs were weighed together. 
Huntingdon Life Sciences 02-5449 Page 25 

Final Report 

2.15.4. TISSUES PRESERVED AND EXAMINED HISTOPATHOLOGICALLY 

TABLE I 

ORGAN NAME 

adrenal glands 

brain 

kidneys 

lungs 

liver 

ovaries 

testes 

gross lesions 

The tissues listed in Table I (page 25) were obtained at sacrifice 

and preserved for all animals. In addition, slides of the lungs were 

prepared and examined microscopically for all animals. Any 

abnormalities not noted during macroscopic examinations which 

were seen during histology processing were recorded. 

EXAMINED 

WEIGHED PRESERVED MICROSCOPICALLY 

X X 

X X 

X X 

X X X 

X X 

X X 

X X 

X 

Preservatives 

All tissues - 10% neutral buffered formalin. 

Testes were placed in Modified Davidson's solution initially and 

then retained in 10% formalin. Lungs were infused with formalin 

prior to their immersion into a larger volume of the same fixative. 

Processing 

After fixation, the lungs from all animals were routinely processed, 

embedded in paraffin, cut at a microtome setting of 4-7 microns, 

mounted on glass slides, stained with hematoxylin and eosin and 

examined by light microscopy. 
Huntingdon Life Sciences 02-5449 

2.16. STATISTICAL ANALYSIS 

Page 26 

Final Report 

A calculation of an estimate of the median lethal concentration, with 95% 

confidence limits was performed for the combined sexes, the males and 

the females for 2-Bromo-3,3,3-Trifluoropropene and E-1-Bromo-3,3,3-

Trifluoropropene, using the method of Litchfield and Wilcoxon. 

Reference for this method is as follows: Litchfield, J.T., Jr. and 

F. Wilcoxon, 1949, J. Pharma. Exp. Therp., 96:99-113. 

2.17. DATA STORAGE 

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

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

the Archives of the Testing Facility upon completion of the study. The 

Sponsor will determine the final disposition of these materials. 

2.18. REGULATORY REFERENCES 

2.18.1. TEST GUIDELINES 

The Organization for Economic Cooperation and Development 

(OECD) Guidelines: Acute Inhalation (12 May 1981), Part 403. 

Health Effects Test Guidelines, Office of Prevention, Pesticides 

and Toxic Substances, United States Environmental Protection 

Agency (OPPTS), August, 1998; OPPTS 870.1300, Acute 

Inhalation Toxicity. 

2.18.2. GOOD LABORATORY PRACTICES 

This study was conducted in compliance with OECD Principles of 

Good Laboratory Practices ENV/MC/CHEM (98) 17 and 

Environmental Protection Agency Good Laboratory Practices as set 

forth in 40 CFR Part 792 (TSCA). 

2.18.3. ANIMAL WELFARE ACT COMPLIANCE 

This study complied with all appropriate parts of the Animal 

Welfare Act regulations: 9 CFR Parts 1 and 2 Final Rules, Federal 

Register, Volume 54, No. 168, August 31, 1989, pp. 36112-36163, 

effective October 30, 1989 and 9 CFR Part 3 Animal Welfare 
Huntingdon Life Sciences 02-5449 Page 27 

Final Report 

Standards; Final Rule, Federal Register, Volume 56, No. 32, 

February 15, 1991, pp. 6426-6505, effective March 18, 1991. 

2.19. PROTOCOL DEVIATIONS 

The following protocol deviation occurred during the study but was not 

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

Since all animals in Group 3 died during the exposure on Study Day 0, the 

chamber was cleared for only 3 minutes rather than the protocol-specified 

30-minute period. 
( 

Huntingdon Life Sciences 02-5449 Page 28 

Final Report 

3. RESULTS AND DISCUSSION 

3.1. CHAMBER MONITORING 

(Table 1) 

The target, analytical (IR) and nominal concentrations for each test group 

are summarized as follows: 

Group Test Substance Target Analytical Nominal 

Concentration Concentration Concentration 

(ppm) (ppm) (ppm) 

1 2-Bromo-3,3,3- 25,000 26,580 29,400 

Trifluoropropene 

2 E-1-Bromo-3,3,3- 25,000 25,650 30,600 

Trifluoropropene 

3 1-Bromo-2- 25,000 24,800 28,700 

Trifluoromethyl-3,3,3-

Trifluoropropene 

4 2-Bromo-3,3,3- 5,000 5,173 6,220 

TrifluorQQropene 

5 E-1-Bromo-3,3,3- 50,000 45,850 52,200 

TrifluorQQTopene 

Particle sizing results demonstrated that all of the exposures were vaporonly since the chamber samples were comparable to room air samples. 

Chamber environmental conditions averaged l7°C temperature and 47% 

relative humidity. 

3.2. MORTALITY 

(Table 6) 

The mortality for each test group is summarized as follows: 

Group Test Substance Analytical Mortality Mortality Mortality 

Concentration Males Females Total 

(ppm) 

1 2-Bromo-3,3,3- 26,580 515 515 10/10 

Trifluoropropene 

4 2-Bromo-3,3,3- 5,173 0/5 0/5 0/10 

Trifluoropropene 

5 E-1-Bromo-3,3,3- 45,850 2/5 0/5 2/10 

Trifluoropropene 

2 E-1-Bromo-3,3,3- 25,650 0/5 0/5 0/10 

Trifluoropro_]J_ene 

3 1-Bromo-2- 24,800 515 515 10/10 

Trifluoromethyl-3,3,3-

Trifluoropropene 
Huntingdon Life Sciences 02-5449 Page 29 

Final Report 

All of the animals exposed to 2-Bromo-3,3,3-Trifluoropropene at 

26,580 ppm were found dead or were euthanized in poor condition on the 

day after the exposure. All of the animals exposed to 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene at 24,800 ppm were 

found dead within 30 minutes of the start of the exposure. Both of the 

male animals exposed to E-1-Bromo-3,3,3-Trifluoropropene at 

45,850 ppm that were found dead died on the day of the exposure. All 

other animals survived to termination. 

Based on the above results, the LC50 values were calculated to be: 

Test Substance 

2-Bromo-3,3,3-

Trifluoropropene 

E-1-Bromo-3,3,3-

Trifluoropropene 

1-Bromo-2-Trifluoromethyl3,3,3-Trifluoropropene 

3.3. BODY WEIGHTS 

(Table 2) 

Animals 

Combined 

Males 

Females 

Combined 

Males 

Females 

Combined 

Males 

Females 

LCSO 95% Confidence Limits 

(ppm) (ppm) 

11726 9054 to 15186 

11726 7584 to 18129 

11726 7584 to 18129 

70955 51449 to 97857 

50950 35358 to 73418 

>45850 Not Calculable 

<24,800 Not Calculable 

<24,800 Not Calculable 

<24,800 Not Calculable 

All surviving animals gained weight (or did not lose weight) during both 

weeks of observations. 

3.4. PHYSICAL OBSERVATIONS 

3.4.1. EXPOSURE PERIOD 

(Table 3) 

The only sign of toxicity noted during the exposure period of 

2-Bromo-3,3,3-Trifluoropropene was labored breathing or gasping 

during the last hour of each exposure. 

The only sign of toxicity noted during the exposure period of 

E-1-Bromo-3,3,3-Trifluoropropene was tail quivering and/or 

tremors during each exposure. The mechanism for this central 

nervous system (CNS) effect was not clear. 
Huntingdon Life Sciences 02-5449 Page 30 

Final Report 

There were no signs of toxicity noted during the exposure period of 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene prior to all 

animals were found dead within 30 minutes of the exposure 

initiation. 

3.4.2. UPON REMOVAL FROM EXPOSURE CHAMBER AND 

OVER THE TWO-HOUR POST-EXPOSURE 

OBSERVATION PERIOD 

(Tables 3 & 5) 

Clinical signs of toxicity noted immediately following the 

exposures of 2-Bromo-3,3,3-Trifluoropropene included clear or red 

nasal discharge, excessive salivation, labored breathing and moist 

rales. The severity and frequency of the signs was increased after 

the higher level exposure of 26,580 ppm compared to the lower 

level exposure of 5,173 ppm. Gasping and decreased activity were 

also noted but only after the higher exposure level. 

Clinical signs of toxicity noted immediately following the 

exposures of E-1-Bromo-3 ,3 ,3-Trifluoropropene included red nasal 

discharge. The severity and frequency of the signs was increased 

after the higher level exposure of 45,850 ppm compared to the 

lower level exposure of 25,650 ppm. Excessive salivation, labored 

breathing and moist rales were also noted but only after the higher 

exposure level. 

3.4.3. DAILY OBSERVATIONS DURING THE 14-DAY POSTEXPOSURE OBSERVATION PERIOD 

(Tables 4 & 5) 

A few signs of toxicity similar to those seen immediately after the 

exposures were noted during the first week of the post -exposure 

observation period. A complete recovery in surviving animals was 

seen during the remainder of the post-exposure observation period. 
Huntingdon Life Sciences 02-5449 

3.5. PATHOLOGY 

3.5.1. ORGAN WEIGHTS 

(Tables 7 & 8) 

Page 31 

Final Report 

There were no apparent effects of exposure on the organs that were 

weighed. This was based on a comparison of the lower level 

exposure of the 2-Bromo-3,3,3-Trifluoropropene to the lower level 

exposure of the E-1-Bromo-3,3,3-Trifluoropropene and to the 

higherlevel exposure of the 1-Bromo-3,3,3-Trifluoropropene. 

3.5.2. MACROSCOPIC 

(Table 9) 

One or more rats from both exposure levels of 2-Bromo-3,3,3-

Trifluoropropene had red discoloration of the lungs. Fluid was 

present in the lungs of one male from the higher level exposure of 

26,580ppm. 

One or more rats from the lower level exposure of 25,650 ppm of 

E-1-Bromo-3,3,3-Trifluoropropene had red discoloration of the 

lungs. Fluid was present in the lungs of two males from the higher 

level exposure of 45,850 ppm. 

One or more rats had red discoloration of the lungs from the 

exposure of 24,800 ppm of 1-Bromo-2-Trifluoromethyl-3,3,3-

Trifluoropropene. 

Microscopically, in the unscheduled deaths, the discoloration was 

due to vascular congestion which is commonly seen in rats not 

exsanguinated (found dead) or inadequately exsanguinated 

(moribund) prior to postmortem examination. The scattered red 

foci were agonal hemorrhages. 

3.5.3. MICROSCOPIC 

(Tables 10 & 11) 

The lungs of all animals on test were routinely processed, cut at 

5 microns, mounted on glass slides, stained with hematoxylin and 

eosin and examined by light microscopy. 
( 

Huntingdon Life Sciences 02-5449 Page 32 

Final Report 

4. 

Test substance-related findings were microscopically present in 

lungs of rats in the higher level exposure of 26,580 ppm of 

2-Bromo-3,3,3-Trifluoropropene. All of the rats, in this group 

only, had bronchiolar lesions consisting of desquamated 

epithelium, bronchiolar/peribronchiolar acute/subacute 

inflammation and/or intraluminal debris; the severity of the 

findings ranged from slight to moderate. Alveolar/intraalveolar 

macrophages (minimal to moderate) normally present in the lungs, 

were most numerous in several of these rats. 

In the rats which were found dead or humanely sacrificed, the 

lungs had vascular congestion. This is a common finding in rats 

which are not exsanguinated (found dead) or incompletely 

exsanguinated (moribund) prior to postmortem examination. 

Therefore, the significance of this finding with respect to the test 

substance is not known. 

Other findings in the lungs occurred sporadically and were not 

considered to be test substance-related; these findings have been 

seen in rats of this strain and age used in similar studies conducted 

in this facility. 

CONCLUSION 

The 4-hour acute inhalation toxicity in rats was compared of 2-Bromo-3,3,3-

Trifluoropropene, E-1-Bromo-3,3,3-Trifluoropropene and 1-Bromo-2-

Trifluoromethyl-3,3,3-Trifluoropropene. On the basis of toxic responses and 

subsequent mortality, E-1-Bromo-3,3,3-Trifluoropropene was considered the least 

toxic with an LC50 of -70,955 ppm, 2-Bromo-3,3,3-Trifluoropropene was 

considered moderately toxic with an LC5o of -11,726 ppm and 1-Bromo-2-

Trifluoromethyl-3,3,3-Trifluoropropene was considered the most toxic with an 

LCso substantially less than the only tested level of 24,800 ppm. 
Huntingdon Life Sciences 

Groups 1 - 3 

House-Line 

Air~= 

Regulator and 

Backpressure 

Gauge 

02-5449 

Diagram of Generation and Sampling Systema 

Glass Bubbler 

with Fritted 

Bottom and 

Test Substance 

Nupro Valve I 

,, ""Hl II ,.=== 

Gen 

Flowmeter 

"'ram1ess Steel "T" 

/ 

Nupro Valve 

Flowmeter 

Stainless Steel 

Strip Chart Recorder 

aSystem enclosed in 10 m3 glass and stainless steel chan]ber. 

Page 33 

Final Report 

Figure 1 

Differential 

Gauge 
Huntingdon Life Sciences 02-5449 Page 34 

Final Report 

Group 4 Diagram of Generation and Sampling Systema Figure 2 

Regulator and 

Backpressure 

Gauge Nupro Valve 

Glass Bubbler r------, 

with F ritted 

Bottom and 

Test Substance 

oil Covered) House-Lin~ ~fi=====;, 

Nitrogen -'<.>' - 9 II r'? 11 Multi meter 

Generation Flowmeter 

Stainless Steel "T" 

Nupro Valve 

House-Line-+ ~!1========'-' 

Air I 

Regulator and 

Back pressure 

Gauge Dilution Flowmeter 

aSystem enclosed in 10 m3 glass and stainless steel chamber. 

ll-+1---------Filter 

Pressure 

Differential 

Gauge 
Huntingdon Life Sciences 

Group 5 

House-Line 

Air -1- Regulator and 

02-5449 

Diagram of Generation and Sampling Systema ----·-··-······-···················-

House-Line 

-1- Nitrogen 

Glass Bubbler with 

Fritted Bottom and 

Test Substance 

(Foil Covered) 

Stainless Steel "T" 

Nupro Valve 

Dilution 

Flowmeter 

Strip Chart Recorder 

asystem enclosed in a 10 m3 glass and stainless steel chamber. 

I 

Pressure 

Differential 

Gauge 

Page 35 

Final Report 

Figure 3 
Huntingdon Life Sciences 02-5449 

Chamber Concentrations, Particle Size 

Page 36 

Final Report 

Grqup 1 Determinations and Chamber Environment Table 1 

Exposure 

Hour 

1 

2 

3 

4 

Mean 

S.D. 

Exposure 

Hour 

Tee 

0.0 

0.5 

1.0 

1.5 

2.0 

2.5 

3.0 

3.5 

4.0 

Mean 

S.D. 

Chamber Concentrations 

Exposure Analytical 

Hour (ppm) 

1 30,400 

2 26,500 

3 24,400 

4 25,000 

Mean 26,580 

S.D. 2,699 

Particle Size Determinations 

MMAD GSD 

{pm) 

0.984 

2.516 

1.264 

2.517 

1.820 

0.812 

1.595 

1.981 

1.789 

2.322 

1.922 

0.310 

Chamber Environment 

Temperature Relative Humidity 

(%) (OC) 

17 

17 

17 

17 

17 

17 

17 

17 

17 

17 

17.0 

0.0 

58 

58 

57 

56 

56 

56 

56 

56 

54 

54 

56.1 

1.4 

1.28E-03 

1.81 E-03 

1.09E-03 

1.39E-03 

1.39E-03 

3.05E-04 

Total 

Chamber Airflow 

(Lpm) 

10.0 

10.0 

10.0 

10.0 

10.0 

10.0 

10.0 

10.0 

10.0 

10.0 

10.0 

0.0 

Key: MMAD = mass median aerodynamic diameter; GSD = geometric standard deviation; 

TMC =total mass concentration; Lpm =Liters per minute. 

a Average room air samples were 1.81 E-03 mg/m3

. 
Huntingdon Life Sciences 02-5449 

Chamber Concentrations, Particle Size 

Group 2 Determinations and Chamber Environment 

Chamber Concentrations 

Exposure Analytical 

Hour (ppm) 

1 27,100 

2 26,200 

3 24,100 

4 25,200 

Mean 25,650 

S.D. 1,292 

Particle Size Determinations 

Exposure MMAD GSD 

Hour (pm) 

1 1.417 2.471 

2 0.932 1.597 

3 0.990 1.712 

4 1.057 1.565 

Mean 1.099 1.836 

S.D. 0.218 0.428 

Chamber Environment 

Exposure 

Page 37 

Final Report 

Table 1 

TMCa 

(mg/m3

) 

1.53E-03 

9.10E-04 

7.03E-04 

6.57E-04 

9.50E-04 

4.02E-04 

Total 

Hour Temperature Relative Humidity Chamber Airflow 

CC) (%) (Lpm) 

Tgg 19 58 10.0 

0.0 19 58 10.0 

0.5 19 58 10.0 

1.0 19 57 10.0 

1.5 20 56 10.0 

2.0 20 56 10.0 

2.5 20 56 10.0 

3.0 20 54 10.0 

3.5 20 53 10.0 

4.0 20 53 10.0 

Mean 19.6 55.9 10.0 

S.D. 0.5 2.0 0.0 

Key: MMAD = mass median aerodynamic diameter; GSD = geometric standard deviation; 

TMC =total mass concentration; Lpm =Liters per minute. 

aAverage room air samples were 1.44E-03 mg/m3

. 
Huntingdon Life Sciences 02-5449 Page 38 

Final Report 

Chamber Concentrations, Particle Size 

Group 3 Determinations and Chamber Environment Table 1 

Exposure 

Hour 

1 

2 

3 

4 

Mean 

S.D. 

Exposure 

Hour 

Tss 

0.0 

0.5 

1.0 

1.5 

2.0 

2.5 

3.0 

3.5 

4.0 

Mean 

S.D. 

Chamber Concentrations 

Exposure Analytical 

Hour (ppm) 

1 

2 

3 

4 

Mean 

S.D. 

24,800 a 

24,800 

Particle Size Determinations 

MMAD GSD 

(pm) 

3.409 1.799 

3.409 1.799 

Chamber Environment 

Temperature Relative Humidity 

(oC) (%) 

19 

19 

19 

19.0 

0.0 

44 

44 

44 

44.0 

0.0 

TMCb 

(mg/m3

) 

9.85E-05 

9.85E-05 

Total 

Chamber Airflow 

(Lpm) 

10.0 

10.0 

10.0 

10.0 

0.0 

Key: MMAD =mass median aerodynamic diameter; GSD =geometric standard deviation; 

TMC =total mass concentration; Lpm = Liters per minute. 

aExposure ended at 33 minutes after all animals were found dead. 

bAverage room air samples were 6.91 E-04 mg/m3

. 
Huntingdon Life Sciences 02-5449 Page 39 

Final Report 

Chamber Concentrations, Particle Size 

Group 4 Determinations and Chamber Environment Table 1 

Exposure 

Hour 

1 

2 

3 

4 

Mean 

S.D. 

Exposure 

Hour 

Tgg 

0.0 

0.5 

1.0 

1.5 

2.0 

2.5 

3.0 

3.5 

4.0 

Mean 

S.D. 

Chamber Concentrations 

Exposure Analytical 

Hour (ppm) 

1 6,250 

2 5,310 

3 4,800 

4 4,330 

Mean 5,173 

S.D. 822 

Particle Size Determinations 

MMAD GSD 

(Jim) 

1.202 1.925 

1.228 1.714 

2.330 2.037 

1.590 1.938 

1.588 1.904 

0.526 0.136 

Chamber Environment 

Temperature Relative Humidity 

(OC) (%) 

16 45 

16 45 

17 44 

17 41 

15 40 

15 40 

15 40 

15 40 

15 40 

15 39 

15.6 41.4 

0.8 2.3 

TMC3 

(mg/m3

) 

8.70E-04 

7.31 E-04 

1.15E-03 

8.85E-04 

9.09E-04 

1.75E-04 

Total 

Chamber Airflow 

(Lpm) 

11.1 

11.1 

11.1 

11.1 

11.1 

11.1 

11.1 

11.1 

11.1 

11.1 

11.1 

0.0 

Key: MMAD = mass median aerodynamic diameter; GSD = geometric standard deviation; 

TMC =total mass concentration; Lpm =Liters per minute. 3

Average room air samples were 1.18E-03 mg/m3

. 
I 

I,. 

Huntingdon Life Sciences 02-5449 

Chamber Concentrations, Particle Size 

Page 40 

Final Report 

Group 5 Determinations and Chamber Environment Table 1 

Chamber Concentrations 

Exposure Analytical 

Hour (ppm) 

1 36,500 

2 39,600 

3 50,400 

4 56,900 

Mean 45,850 

S.D. 9,474 

Particle Size Determinations 

Exposure MMAD GSD TMCa 

Hour (Jim) (mg/m3

) 

1 0.975 1.533 8.08E-04 

2 0.943 1.480 6.03E-04 

3 1.252 1.769 8.48E-04 

4 1.116 1.519 7.89E-04 

Mean 1.072 1.575 7.62E-04 

S.D. 0.142 0.131 1.09E-04 

Chamber Environment 

Exposure Total 

Hour Temperature Relative Humidity Chamber Airflow 

CC) (%) (Lpm) 

Tss 16 46 6.5 

0.0 16 45 6.5 

0.5 16 44 6.5 

1.0 16 41 6.5 

1.5 16 36 6.5 

2.0 16 36 6.5 

2.5 16 34 6.5 

3.0 16 36 6.5 

3.5 16 38 6.5 

4.0 16 38 6.5 

Mean 16.0 39.4 6.5 

S.D. 0.0 4.3 0.0 

Key: MMAD = mass median aerodynamic diameter; GSD = geometric standard deviation; 

TMC =total mass concentration; Lpm = Liters per minute. 

aAverage room air samples were 2.24E-03 mg/m3

. 
Huntingdon Life Sciences 02-5449 

Body Weights and Body Weight Changes 

M9-les from Day 0 

Page 41 

Final Report 

Table 2 

Animal 

Number Body Weight (grams) 

Body Weight Change 

from Day 0 (grams) 

DAY 0 DAY 7 DAY 14 DAY 7 DAY 14 

Group 1 - 2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

1 046 275 AD AD 

1047 262 AD AD 

1 048 263 AD AD 

1 049 256 AD AD 

1 050 273 AD AD 

Mean 266 

S.D. 8.0 

Group 2- E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

2046 276 319 377 43 101 

204 7 280 340 407 60 127 

2048 268 326 388 58 120 

2049 260 315 366 55 106 

2050 254 291 329 37 75 

Mean 268 318 373 51 1 06 

S.D. 10.8 17.9 29.1 10.1 20.1 

Group 3- 1-Bromo-2-Trifluoromethyi-3,3,3-Trifluoropropene (24,800 ppm) 

3046 264 AD AD 

304 7 263 AD AD 

3048 252 AD AD 

3049 269 AD AD 

3050 260 AD AD 

Mean 262 

S.D. 6.3 

Group 4 - 2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

4046 240 269 335 29 95 

4047 239 275 337 36 98 

4048 241 273 341 32 100 

4049 236 248 318 12 82 

4050 266 306 373 40 1 07 

Mean 244 274 341 30 96 

S.D. 12.2 20.8 20.0 10.8 9.2 

Group 5- E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

5046 240 306 367 66 127 

5047 242 AD AD 

5048 239 300 358 61 119 

5049 240 AD AD 

5050 227 278 314 51 87 

Mean 238 295 346 59 111 

S.D. 6.0 14.7 28.4 7.6 21.2 

AD =Animal Dead. 
1·. 

Huntingdon Life Sciences 02-5449 

Body Weights and Body Weight Changes 

Females from Day 0 

Page 42 

Final Report 

Table 2 

Animal 

Number Body Weight (grams) 

BODY WEIGHT CHANGE 

from Day 0 (grams) 

DAY 0 DAY 7 DAY 14 DAY7 DAY14 

Group 1 - 2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

1546 196 AD AD 

1547 215 AD AD 

1548 202 AD AD 

1549 203 AD AD 

1550 204 AD AD 

Mean 204 

S.D. 6.9 

Group 2- E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

2546 187 199 205 12 

2547 212 222 246 10 

2548 199 218 235 19 

2549 205 223 231 18 

2550 207 228 256 21 

Mean 202 218 235 16 

S.D. 9.6 11.2 19.2 4.7 

18 

34 

36 

26 

49 

33 

11.6 

Group 3 -1-Bromo-2-Trifluoromethyi-3,3,3-Trifluoropropene (24,800 ppm) 

3546 181 AD AD 

3547 203 AD AD 

3548 199 AD AD 

3549 187 AD AD 

3550 204 AD AD 

Mean 195 

S.D. 10.3 

Group 4- 2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

4546 218 242 280 24 62 

4547 212 221 232 9 20 

4548 209 223 247 14 38 

4549 225 231 247 6 22 

4550 221 221 248 0 27 

Mean 217 228 251 11 34 

S.D. 6.5 9.0 17.6 9.0 17.2 

Group 5- E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

5546 205 216 234 11 29 

5547 203 221 236 18 33 

5548 209 220 238 11 29 

5549 217 226 237 9 20 

5550 211 218 247 7 36 

Mean 209 220 238 11 29 

S.D. 5.5 3.8 5.0 4.1 6.0 

AD =Animal Dead. 
(• .. 

\ 

Huntingdon Life Sciences 02-5449 

Summary of Day 0 Observations 

HOURS INTO EXPOSUREa 

PRETESTb 1/4 1/2 3/4 1 2 3 4 

M/F 

Group 1 - 2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

No. of Animals Observed 5/5 10 10 10 10 10 10 10 

NORMAL 

Within Normal Limits 5/5 10 10 10 10 10 10 5 

SECRETORY 

Clear Nasal Discharge 0/0 0 0 0 0 0 0 0 

Excessive Salivation 0/0 0 0 0 0 0 0 0 

RESPIRATORY 

Labored Breathing 0/0 0 0 0 0 0 0 5 

Gasping 0/0 0 0 0 0 0 0 0 

Moist Rales 0/0 0 0 0 0 0 0 0 

GENERAL 

Decreased Activity 0/0 0 0 0 0 0 0 0 

T* =Animals were observed upon removal from the chamber. 

a Animals were observed as a group. 

b Animals were observed individually. Number represents animals exhibiting signs. 

M = Male, F = Female 

Page 43 

Final Report 

Table 3 

HOURS 

POSTEXPOSUREb 

T* 1 2 

M/F M/F M/F 

5/5 5/5 5/5 

0/0 0/0 0/0 

4/4 5/4 5/5 

3/2 3/2 3/2 

3/0 3/1 4/1 

1/0 1/0 1/0 

1/4 1/4 0/4 

3/0 4/0 4/0 
I' I . \ 

Huntingdon Life Sciences 02-5449 

Summary of Day 0 Observations 

HOURS INTO EXPOSURE3 

PRETESTb 1/4 1/2 3/4 1 2 3 

M/F 

Group 2- E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

No. of Animals Observed 5/5 10 10 10 10 10 10 

NORMAL 

Within Normal Limits 5/5 3 3 3 3 3 3 

SECRETORY 

Red Nasal Discharge 0/0 0 0 0 0 0 0 

GENERAL 

Quivering Tail 0/0 7 7 7 7 7 7 

Group 3 -1-Bromo-2-Trifluoromethyi-3,3,3-Trifluoropropene (24,800 ppm) 

No. of Animals Observed 5/5 1 0 1 0 0° 

NORMAL 

Within Normal Limits 

DEAD 

Found Dead 

5/5 0 0 

0/0 10 10 

4 

10 

8 

0 

2 

T* = Animals were observed upon removal from the chamber. 

a Animals were observed as a group. 

b Animals were observed individually. Number represents animals exhibiting signs. 

M = Male, F = Female 

cExposure ended at 33 minutes after all animals were found dead. 

Page 44 

Final Report 

Table 3 

HOURS 

POSTEXPOSUREb 

T* 1 2 

M/F M/F M/F 

5/5 5/5 5/5 

1/4 2/5 2/5 

4/1 3/0 3/0 

0/0 0/0 0/0 
I 

Huntingdon Life Sciences 02-5449 

Summary of Day 0 Observations 

HOURS INTO EXPOSUREa 

PRETESTb 1/4 1/2 3/4 1 2 3 4 

M/F 

Group 4- 2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

No. of Animals Observed 5/5 10 10 10 10 10 10 10 

NORMAL 

Within Normal Limits 5/5 10 10 10 10 10 10 9 

SECRETORY 

Red Nasal Discharge 0/0 0 0 0 0 0 0 0 

Red Dried Material on Face 0/0 0 0 0 0 0 0 0 

RESPIRATORY 

Labored Breathing 0/0 0 0 0 0 0 0 0 

Moist Rales 0/0 0 0 0 0 0 0 0 

Gasping 0/0 0 0 0 0 0 0 1 

T* =Animals were observed upon removal from the chamber. 

a Animals were observed as a group. 

b Animals were observed individually. Number represents animals exhibiting signs. 

M = Male, F = Female 

Page 45 

Final Report 

Table 3 

HOURS 

POSTEXPOSUREb 

T* 1 2 

M/F M/F M/F 

5/5 5/5 5/5 

2/3 2/3 2/3 

1/1 0/1 0/1 

2/0 0/0 0/0 

0/1 0/1 0/1 

2/0 3/0 3/0 

0/0 0/0 0/0 
Huntingdon Life Sciences 02-5449 

Summary of Day 0 Observations 

HOURS INTO EXPOSUREa 

PRETESTb 1/4 1/2 3/4 

M/F 

Group 5- E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

No. of Animals Observed 5/5 10 10 10 

NORMAL 

Within Normal Limits 5/5 5 5 5 

SECRETORY 

Lacrimation 0/0 0 0 0 

Clear Nasal Discharge 0/0 0 0 0 

Red Nasal Discharge 0/0 0 0 0 

Excessive Salivation 0/0 0 0 0 

Chromodacryorrhea 0/0 0 0 0 

RESPIRATORY 

Labored Breathing 0/0 0 0 0 

Moist Rales 0/0 0 0 0 

GENERAL 

Tremors!rail Quivering 0/0 5 5 5 

DEAD 

Found Dead 0/0 0 0 0 

. T* = Animals were observed upon removal from the chamber. 

a Animals were observed as a group. 

1 2 3 4 

10 10 10 10 

5 5 4 4 

0 0 1 2 

0 0 0 0 

0 0 0 0 

0 0 0 0 

0 0 0 0 

0 0 0 0 

0 0 0 0 

5 5 5 4 

0 0 0 1 

b Animals were observed individually. Number represents animals exhibiting signs. 

M = Male, F = Female 

Page 46 

Final Report 

Table 3 

HOURS 

POSTEXPOSUREb 

T* 1 2 

M/F M/F M/F 

3/5 3/5 3/5 

0/2 0/2 0/2 

0/1 0/1 0/1 

0/0 0/0 1/0 

2/2 3/2 2/2 

2/1 1/1 0/1 

0/1 0/1 0/1 

1/0 1/0 1/1 

1/1 1/1 1/1 

0/0 0/0 0/0 

1/0 0/0 0/0 
Huntingdon Life Sciences 02-5449 

Summary of 14-Day Post-Exposure 

Observations 

Da 

1 2 3 4 5 6 7 8 

Group 1 - 2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

Males 

No. of Animals Examined 5 0 0 0 0 0 0 0 

Within Normal Limits 0 0 0 0 0 0 0 0 

SECRETORY 

Clear Nasal Discharge 4 0 0 0 0 0 0 0 

Excessive Salivation 1 0 0 0 0 0 0 0 

Chromodacryorrhea 2 0 0 0 0 0 0 0 

RESPIRATORY 

Labored Breathing 3 0 0 0 0 0 0 0 

Gasping 1 0 0 0 0 0 0 0 

GENERAL 

Decreased Activity 3 0 0 0 0 0 0 0 

Yellow Ano-Genital Staining 1 0 0 0 0 0 0 0 

DEAD 

Found Dead 1 0 0 0 0 0 0 0 

Humane Sacrifice 4 0 0 0 0 0 0 0 

Females 

No. of Animals Examined 5 0 0 0 0 0 0 0 

Within Normal Limits 0 0 0 0 0 0 0 0 

SECRETORY 

Clear Nasal Discharge 5 0 0 0 0 0 0 0 

Excessive Salivation 3 0 0 0 0 0 0 0 

Dried Red Material Facial Area 1 0 0 0 0 0 0 0 

RESPIRATORY 

Gasping 3 0 0 0 0 0 0 0 

Moist Rales 2 0 0 0 0 0 0 0 

GENERAL 

Yellow Ano-Genital Staining 2 0 0 0 0 0 0 0 

DEAD 

Found Dead 1 0 0 0 0 0 0 0 

Humane Sacrifice 4 0 0 0 0 0 0 0 

9 10 11 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

Page 47 

Final Report 

Table 4 

12 13 14 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 
Huntingdon Life Sciences 02-5449 

Summary of 14-Day Post-Exposure 

ObseNations 

Da 

1 2 3 4 5 6 7 8 9 

Group 2- E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

Males 

No. of Animals Examined 5 5 5 5 5 5 5 5 5 

Within Normal Limits 5 5 5 5 5 5 5 5 5 

Females 

No. of Animals Examined 5 5 5 5 5 5 5 5 5 

Within Normal Limits 5 5 5 5 5 5 5 5 5 

Group 3- 1-Bromo-2-Trifluoromethyi-3,3,3-Trifluoropropene (24,800 ppm) 

Males 

No. of Animals Examineda 0 0 0 0 0 0 0 0 0 

Females 

No. of Animals Examineda 0 0 0 0 0 0 0 0 0 

aAnimals died during exposure. 

10 11 

5 5 

5 5 

5 5 

5 5 

0 0 

0 0 

Page 48 

Final Report 

Table 4 

12 13 14 

5 5 5 

5 5 5 

5 5 5 

5 5 5 

0 0 0 

0 0 0 
Huntingdon Life Sciences 02-5449 

Summary of 14-Day Post-Exposure 

Observations 

Da 

1 2 3 4 5 6 7 8 

Group 4- 2-Bromo-3,3,3-Trifluoropropene (5,173 ppm) 

Males 

No. of Animals Examined 5 5 5 5 5 5 5 5 

Within Normal Limits 0 0 1 4 3 3 4 4 

RESPIRATORY 

Gasping 1 0 0 0 0 0 0 0 

Moist Rales 3 4 4 1 2 2 1 1 

GENERAL 

Decreased Fecal Volume 5 5 0 0 0 0 0 0 

Females 

No. of Animals Examined 5 5 5 5 5 5 5 5 

Within Normal Limits 0 2 2 4 2 5 5 4 

SECRETORY 

Red Nasal Discharge 0 1 1 0 0 0 

RESPIRATORY 

Labored Breathing 1 0 0 0 0 0 0 0 

Moist Rales 2 3 3 0 1 0 0 0 

GENERAL 

Alopecia- Extremities/Snout 0 0 0 0 1 0 0 0 

No Stool 1 0 0 0 0 0 0 0 

Decreased Fecal Volume 4 1 0 0 0 0 0 0 

Incisors Maloccluded 0 0 0 0 0 0 0 1 

9 10 11 

5 5 5 

5 5 5 

0 0 0 

0 0 0 

0 0 0 

5 5 5 

4 4 4 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

1 1 1 

Page 49 

Final Report 

Table 4 

12 13 14 

5 5 5 

5 5 5 

0 0 0 

0 0 0 

0 0 0 

5 5 5 

4 4 4 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

0 0 0 

1 1 1 
Huntingdon Life Sciences 02-5449 

Summary of 14-Day Post-Exposure 

ObseNations 

Da 

1 2 3 4 5 6 7 8 

Group 5- E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

Males 

No. of Animals Examined 3 3 3 3 3 3 3 3 

Within Normal Limits 2 2 3 3 3 3 3 3 

RESPIRATORY 

Labored Breathing 1 0 0 0 0 0 0 

Females 

No. of Animals Examined 5 5 5 5 5 5 5 5 

Within Normal Limits 3 5 5 5 5 5 5 5 

SECRETORY 

Red Nasal Discharge 2 0 0 0 0 0 0 0 

RESPIRATORY 

Labored Breathing 1 0 0 0 0 0 0 0 

9 10 11 

3 3 3 

3 3 3 

0 0 0 

5 5 5 

5 5 5 

0 0 0 

0 0 0 

Page 50 

Final Report 

Table 4 

12 13 14 

3 3 3 

3 3 3 

0 0 0 

5 5 5 

5 5 5 

0 0 0 

0 0 0 
Huntingdon Life Sciences 02-5449 

Males I Individual Observations 

Animal 

Number Observations 1 2 3 

Pre T* 1 2 

Group 1 - 2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

1046 Within Normal Limits p 

Clear Nasal Discharge p p p p 

Excessive Salivation p p p 

Chromodacryorrhea p 

Labored Breathing p p 

Decreased Activity p p p p 

Humane Sacrifice p 

1047 Within Normal Limits p 

Clear Nasal Discharge p p p p 

Excessive Salivation p p p p 

Chromodacryorrhea p 

Labored Breathing p p p p 

Moist Rales s s 

Decreased Activity p p p p 

Humane Sacrifice p 

1048 Within Normal Limits p 

Clear Nasal Discharge p p p p 

Gasping p p p p 

Yellow Ana-Genital Staining s 

Humane Sacrifice p 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 

I 

Da 

4 5 6 7 8 9 10 11 

Table 5 

Page 51 

Final Reoort 

12 13 14 
Huntingdon Life Sciences 02-5449 Page 52 

Final Reoort 

I Males I Individual Observations · I Table 5 I 

Animal Day 

Number Observations 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 

Pre T* 1 2 

1049 Within Normal Limits p 

Clear Nasal Discharge p p p p 

Labored Breathing p p p p 

Decreased Activity p p p p 

Humane Sacrifice p 

1050 Within Normal Limits p 

Clear Nasal Discharge p p 

Excessive Salivation p p p 

Labored Breathing p p p 

Decreased Activity p p 

Found Dead p 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 
Huntingdon Life Sciences 02-5449 Page 53 

Final Reoort 

I ~::es I Individual Observations I Table 5 I 

Animal Day: 

Number Observations 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 

Pre T* 1 2 

Group 2- E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

2046 Within Normal Limits p p p p p p p p p p p p p p p 

Red Nasal Discharge p p p 

2047 Within Normal Limits p p p p p p p p p p p p p p p 

Red Nasal Discharge p p p 

2048 Within Normal Limits p p p p p p p p p p p p p p p 

Red Nasal Discharge p p p 

2049 Within Normal Limits p p p p p p p p p p p p p p p p p 

Red Nasal Discharge p 

2050 Within Normal Limits p p p p p p p p p p p p p p p p p p 

Group 3- 1-Bromo-2-Trifluoromethyi-3,3,3-Trifluoropropene (24,800 ppm) 

3046 Within Normal Limits p 

Died During Exposure p 

3047 Within Normal Limits p 

Died During Exposure p 

3048 Within Normal Limits p 

Died During Exposure p 

3049 Within Normal Limits p 

Died During Exposure p 

3050 Within Normal Limits p 

Died During Exposure p 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 
Huntingdon Life Sciences 02-5449 Page 54 

I Males I Individual ObseNations I Table 5 I 

Animal Day: 

Number Observations 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 

Pre T* 1 2 

Group 4- 2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

4046 Within Normal Limits p p p p p p p p p p p p 

Red Nasal Discharge p 

Red Dried Material on Face s 

Moist Rales s s s s s 

Decreased Fecal Volume p p 

4047 Within Normal Limits p p p p p p p p p p p p p p p p 

Decreased Fecal Volume p p 

4048 Within Normal Limits p p p p p p p p p p 

Moist Rales s s s s s s s s 

Decreased Fecal Volume p p 

4049 Within Normal Limits p p p p p p p p p p 

Gasping p 

Moist Rales s s s s s s s 

Decreased Fecal Volume p p 

4050 Within Normal Limits p p p p p p p p p p p p 

Red Dried Material on Face s 

Moist Rales s s s M s s 

Decreased Fecal Volume p p 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 
Huntingdon Life Sciences 02-5449 Page 55 

I Males I Individual Observations I Table 5 I 

Animal Day: 

Number Observations 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 

Pre T* 1 2 

Group 5- E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

5046 Within Normal Limits p p p p p p p p p p p p p 

Red Nasal Discharge p p p 

Excessive Salivation p p 

Labored Breathing p p p p p 

5047 Within Normal Limits p 

Found Dead During Exposure p 

5048 Within Normal Limits p p p p p p p p p p p p p p p 

Red Nasal Discharge p p p 

Excessive Salivation p 

5049 Within Normal Limits p 

Dead p 

5050 Within Normal Limits p p p p p p p p p p p p p p p 

Clear Nasal Discharge p 

Red Nasal Discharge p 

Moist Rales s s s 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 
Huntingdon Life Sciences 02-5449 

Females I Individual Observations 

Animal 

Number Observations 0 1 2 3 

Pre T* 1 2 

Group 1 - 2-Bromo-3,3,3-Trifluoropropene {26,580 ppm) 

1546 Within Normal Limits p 

Clear Nasal Discharge p p p p 

Red Dried Material on Face s 

Gasping p 

Moist Rales M M s 

Humane Sacrifice p 

1547 Within Normal Limits p 

Clear Nasal Discharge p p p p 

Excessive Salivation p 

Labored Breathing p p 

Gasping p 

Humane Sacrifice p 

1548 Within Normal Limits p 

Clear Nasal Discharge p p 

Excessive Salivation p 

Moist Rales s s s s 

Yellow Ana-Genital Staining s 

Found Dead p 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 

I 

Da ----------------· ··-- ··-··-·-· ---

4 5 6 7 8 9 10 11 

Table 5 

Page 56 

Final Renort 

12 13 14 
Huntingdon Life Sciences 02-5449 Page 57 

Eioal BepQ!:t 

Females I Individual Observations I Table 5 

Animal Da 

Number Observations 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 

Pre T* 1 2 

1549 Within Normal Limits p 

Clear Nasal Discharge p p p p 

Excessive Salivation p p p p 

Gasping p 

Moist Rales s s s 

Yellow Ana-Genital Staining s 

Humane Sacrifice p 

1550 Within Normal Limits p 

Clear Nasal Discharge p p p p 

Excessive Salivation p p p 

Moist Rales s s s 

Humane Sacrifice p 

Group 2- E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

2546 Within Normal Limits p p p p p p p p p p p p p p p p p p 

2547 Within Normal Limits p p p p p p p p p p p p p p p p p p 

2548 Within Normal Limits p p p p p p p p p p p p p p p p p p 

2549 Within Normal Limits p p p p p p p p p p p p p p p p p p 

2550 Within Normal Limits p p p p p p p p p p p p p p p p p 

Red Nasal Discharge p 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 
Huntingdon Life Sciences 02-5449 

Females Individual Observations Table 5 

Dav 

Page 58 

Final Reoort 

Animal 

Number Observations 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 

Pre T* 1 2 

Group 3 - 1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene (24,800 ppm) 

3546 Within Normal Limits p 

Died During Exposure p 

3547 Within Normal Limits p 

Died During Exposure p 

3548 Within Normal Limits p 

Died During Exposure p 

3549 Within Normal Limits p 

Died During Exposure p 

3550 Within Normal Limits p 

Died During Exposure p 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 
Huntingdon Life Sciences 02-5449 Page 59 

Eioal Bepm:t 

Females I Individual Observations I Table 5 I 

Animal Day ___ 

Number Observations 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 

Pre T* 1 2 

Group 4- 2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

4546 Within Normal Limits p p p p p p p p p p p p p p p p p 

Decreased Fecal Volume p 

4547 Within Normal Limits p p p p p p p p p p p p p p p p 

Alopecia- Extremities/Snout M 

Decreased Fecal Volume p 

4548 Within Normal Limits p p p p p p p p p p 

Red Nasal Discharge p p p p 

Labored Breathing p p p p 

Moist Rales s s 

No Stool p 

Decreased Fecal Volume p 

4549 Within Normal Limits p p p p 

Red Nasal Discharge p p p 

Moist Rales M M s s 

Decreased Fecal Volume p p 

Incisors Maloccluded p p p p p p p 

4550 Within Normal Limits p p p p p p p p p p p p p p p 

Moist Rales s s s 

Decreased Fecal Volume p 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 
Huntingdon Life Sciences 02-5449 

Females I Individual Observations 

Animal 

Number Observations 0 1 2 3 

Pre T* 1 2 

Group 5- E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

5546 Within Normal Limits p p p p 

Moist Rales s s s 

5547 Within Normal Limits p p p 

Red Nasal Discharge p p p p 

Chromodacryorrhea p p p 

5548 Within Normal Limits p p p p p p p 

5549 Within Normal Limits p p p 

Lacrimation M M M 

Red Nasal Discharge p p p p 

Excessive Salivation p p p 

Labored Breathing p p 

5550 Within Normal Limits p p p p p p p 

Key: P = Present; S = Slight; M = Moderate; E = Extreme. 

I 

Da 

4 5 6 7 8 

p p p p p 

p p p p p 

p p p p p 

p p p p p 

p p p p p 

9 10 11 

p p p 

p p p 

p p p 

p p p 

p p p 

Table 5 

Page 60 

Final Reoort 

12 13 14 

p p p 

p p p 

p p p 

p p p 

p p p 
Huntingdon Life Sciences 02-5449 

Animal Termination History 

Animal Date of Type of Study Animal Date of Type of 

Number Death Death3 Dal Number Death Death a 

Males Females 

Group 1 - 2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

1046 18 Sep 02 H 1 1546 18 Sep 02 H 

1047 18 Sep 02 H 1 1547 18 Sep 02 H 

1048 18 Sep 02 H 1 1548 18 Sep 02 F 

1049 18 Sep 02 H 1 1549 18 Sep 02 H 

1050 18 Sep 02 F 1 1550 18 Sep 02 H 

Group 2- E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

2046 01 Oct 02 T 14 2546 01 Oct 02 T 

2047 01 Oct 02 T 14 2547 01 Oct 02 T 

2048 01 Oct 02 T 14 2548 01 Oct 02 T 

2049 01 Oct 02 T 14 2549 01 Oct 02 T 

2050 01 Oct 02 T 14 2550 01 Oct 02 T 

Group 3- 1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene (24,800 ppm) 

3046 17 Sep 02 F 0 3546 17 Sep 02 F 

3047 17 Sep 02 F 0 3547 17 Sep 02 F 

3048 17 Sep 02 F 0 3548 17 Sep 02 F 

3049 17 Sep 02 F 0 3549 17 Sep 02 F 

3050 17 Sep 02 F 0 3550 17 Sep 02 F 

Group 4- 2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

4046 04 Nov 02 T 14 4546 04 Nov02 T 

4047 04 Nov 02 T 14 4547 04 Nov 02 T 

4048 04 Nov 02 T 14 4548 04 Nov 02 T 

4049 04 Nov02 T 14 4549 04 Nov02 T 

4050 04 Nov 02 T 14 4550 04 Nov02 T 

Group 5- E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

5046 04 Nov 02 T 14 5546 04 Nov 02 T 

5047 21 Oct 02 F 0 5547 04 Nov 02 T 

5048 04 Nov 02 T 14 5548 04 Nov02 T 

5049 21 Oct 02 F 0 5549 04 Nov 02 T 

5050 04 Nov02 T 14 5550 04 Nov 02 T 

aH =Humane Sacrifice; F =Found Dead; T =Terminal Sacrifice. 

bDay of exposure is defined as Day 0. 

Page 61 

Final Report 

Table 6 

Study 

Dal 

1 

1 

1 

1 

1 

14 

14 

14 

14 

14 

0 

0 

0 

0 

0 

14 

14 

14 

14 

14 

14 

14 

14 

14 

14 
Huntingdon Life Sciences 02-5449 Page 62 

Final Report 

Mean Organ Weights 

Preface Table 7 

Table of Contents 

Absolute Organ Weights ..................................................................................... 63 

%Organ to Body Weight Ratios ......................................................................... 65 

%Organ to Brain Weight Ratios ......................................................................... 67 

Key to Abbreviations: 

g 

wt. 

observ. 

= Grams 

= Weight 

= Observed 

Corresponding exposure levels for each group were as follows: 

Group 1 

Group 2 

Group 3 

Group 4 

Group 5. 

Note: 

2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

1-Bromo-2-Trifluoromethyi-3,3,3-Trifluoropropene (24,800 ppm) 

2-Bromo-3,3,3-Trifluoropropene (5,173 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

Organs were not weighed for Groups 1 and 3 because all animals were found 

dead or humanely sacrificed. 
Huntingdon Life Sciences 

Princeton Research Center 

East Millstone, New Jersey 

Terminal Sacrifice 

Group 

2 

M e a n: 

Standard deviation: 

Number of observ. : 

4 

Me an: 

Standard deviation: 

Number of observ. : 

5 

M e a n: 

Standard deviation: 

Number of observ. : 

Terminal 

Body wt. (g) 

373.4 

29.1 

(5) 

340.8 

20.0 

(5) 

346.3 

28.4 

(3) 

Summary statistics for absolute organ weights (g) 

Study number: 025449 

Brain Liver 

Adrenal Glands Kidneys 

M a 1 e A n i m a 1 s 

0.0637 2.2003 3.5118 17.2573 

0.0051 0.1439 0.0758 1. 3656 

(5) (5) (5) (5) 

0.0571 1.9022 2.9483 14.5060 

0.0089 0.0702 0.2360 0.6510 

(5) (5) (5) (5) 

0.0499 2. 0213 3.1764 15.2994 

0.0067 0.0657 0.3363 2.3093 

(3) (3) (3) (3) 

Printed: 08-Jan-03 

Version 4.2.2 

Testes 

Lungs 

1.7918 3.3099 

0.1113 0.2281 

(5) (5) 

1.4642 3.0933 

0.0919 0.1254 

(5) (5) 

1.5696 3.0283 

0.3246 0.1112 

(3) (3) 

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Huntingdon Life Sciences 

Princeton Research Center 

East Millstone, New Jersey 

Terminal Sacrifice 

Group 

2 

M e a n: 

Standard deviation: 

Number of observ. : 

4 

M e a n: 

Standard deviation: 

Number of observ. : 

5 

M e a n: 

Standard deviation: 

Number of observ. : 

Terminal 

Body wt. (g) 

234.6 

19.2 

(5) 

250.8 

17.6 

(5) 

238.4 

5.0 

(5) 

Summary statistics for absolute organ weights (g) 

Study number: 025449 

Brain Liver 

Adrenal Glands Kidneys 

F e m a l e A n i m a l s 

0.0716 1.9653 2.3739 10.1943 

0.0049 0.0622 0.1854 1. 2445 

(5) (5) (5) (5) 

0.0760 1.8957 2.1833 11.1084 

0.0058 0.0833 0.1977 2.4203 

(5) (5) (5) (5) 

0.0675 1. 9013 2.0216 10.6981 

0.0067 0.0658 0.1218 0.2301 

(5) (5) (5) (5) 

Printed: 08-Jan.-03 

Version 4.2.2 

ovaries 

Lungs 

1.4059 0.0935 

0.0794 0.0056 

(5) (5) 

1.4306 0.1020 

0.2357 0.0152 

(5) (5) 

1.3208 0 .. 0945 

0.0602 0.0139 

(5) (5) 

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Huntingdon Life Sciences Summary Statistics for % Organ to Body Weight Printed: 08-Jan-03 

Princeton Research Center Study number: 025449 

East Millstone, New Jersey Version 4.2.2 

Terminal Sacrifice 

Group Terminal Brain Liver Testes 

Body wt. (g) Adrenal Glands Kidneys Lungs 

---------------------------------------------------------------------------------------------------------------------~~------------

M a l e A n i m a l s 

2 

M e a n: 373.4 0.0171 0.5940 0.9461 4.6265 0.4811 0.8937 

Standard deviation: 29.1 0. 0014 0.0792 0. 0918 0.2528 0. 0296 0.1226 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

4 

Me an: 340.8 0.0168 0.5601 0.8648 4.2623 0.4319 0.9089 

Standard deviation: 20.0 0.0026 0.0447 0.0403 0.1936 0.0502 0.0380 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

5 

M e a n: 346.3 0.0144 0.5862 0. 9178 4.4028 0.4504 0.8789 

Standard deviation: 28.4 0.0014 0.0496 0. 0726 0.3715 0.0639 0.0866 

Number of observ. : (3) (3) (3) (3) (3) (3) (3) 

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Huntingdon Life Sciences Summary Statistics for % Organ to Body Weight Printed: 08-Jan-03 

Princeton Research Center Study number: 025449 

East Millstone, New Jersey Version 4.2.2 

Terminal Sacrifice 

Group Terminal Brain Liver ovaries 

Body wt. (g) Adrenal Glands Kidneys Lungs 

-----------------------------------------------------------------------------------------------------------------------~-----------

F e m a l e A n i m a l s 

2 

M e a n: 234.6 0.0308 0.8421 1. 0154 4.3368 0.6005 0.0400 

Standard deviation: 19.2 0.0038 0. 0727 0.0880 0.2318 0.0222 0.0036 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

4 

M e a n: 250.8 0.0304 0.7582 0.8698 4.3952 0.5706 0.0408 

Standard deviation: 17.6 0.0030 0.0529 0.0274 0.6150 0.0884 0.0063 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

5 

M e a n: 238.4 0.0283 0.7977 0.8481 4.4903 0.5541 0.0397 

Standard deviation: 5.0 0.0026 0.0300 0.0508 0.1791 0.0256 0.0062 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

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Huntingdon Life Sciences 

Princeton Research Center 

East Millstone, New Jersey 

Terminal Sacrifice 

Group Terminal 

Body wt. (g) 

2 

M e a n: 373.4 

Standard deviation: 29.1 

Number of observ. : (5) 

4 

M e a n: 340.8 

Standard deviation: 20.0 

Number of observ. : (5) 

5 

M e a n: 346.3 

Standard deviation: 28.4 

Number of observ. : (3) 

Summary Statistics for % Organ to Brain Weight 

Study number: 025449 

Brain Liver 

Adrenal Glands Kidneys 

M a 1 e A n i m a 1 s 

2.9061 100.0000 160.0990 787.7351 

0.3013 0.0000 10.0132 88.5729 

(5) (5) (5) (5) 

3.0182 100.0000 155.1873 763.1697 

0.5697 0.0000 14.2520 38.8130 

(5) (5) (5) (5) 

2.4749 100.0000 156.9516 756.1699 

0.3896 0.0000 12.3757 103.9140 

(3) (3) (3) (3) 

Printed: 08-Jan-03 

Version 4.2.2 

Testes 

Lungs 

81.9524 150.5403 

10.3061 7.5646 

(5) (5) 

77.0101 162.9808 

4.7813 12.5995 

(5) (5) 

77.5256 149.8154 

14.8739 2.0838 

(3) (3) 

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Huntingdon Life Sciences 

Princeton Research Center 

East Millstone, New Jersey 

Terminal Sacrifice 

Group Terminal 

Body wt. (g) 

2 

M e a n: 234.6 

Standard deviation: 19.2 

Number of observ. : (5) 

4 

M e a n: 250.8 

Standard deviation: 17.6 

Number of observ. : (5) 

5 

M e a n: 238.4 

Standard deviation: 5.0 

Number of observ. : (5) 

Summary Statistics for % Organ to Brain Weight 

Study number: 025449 

Brain Liver 

Adrenal Glands Kidneys 

F e m a 1 e A n i m a 1 s 

3.6530 100.0000 121.0075 518.2682 

0.3581 0.0000 11.8241 55.0725 

(5) (5) (5) (5) 

4.0194 100.0000 115.1597 585.5146 

0.3796 0.0000 8.6870 118.8030 

(5) (5) (5) (5) 

3.5583 100.0000 106.5000 563.2047 

0.4511 0.0000 8.6047 23.0241 

(5) (5) (5) (5) 

Printed: 08-Jan-03 

Version 4.2.2 

Ovaries 

Lungs 

71.5945 4.7534 

4.5674 0.1722 

(5) (5) 

75.6661 5.4022 

13.4477 0.9180 

(5) (5) 

69.5197 4.9744 

3.5674 0.7266 

(5) (5) 

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Huntingdon Life Sciences 

Table of Contents 

02-5449 

Individual Organ Weights 

Preface 

Page 69 

Final Report 

Table 8 

Absolute Organ Weights ..................................................................................... 70 

%Organ to Body Weight Ratios ......................................................................... 72 

%Organ to Brain Weight Ratios ......................................................................... 74 

Key to Abbreviations: 

g 

wt. 

observ. 

= Grams 

= Weight 

= Observed 

Corresponding exposure levels for each group were as follows: 

Group 1 

Group 2 

Group 3 

Group 4 

Group 5 

Note: 

2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene (24,800 ppm) 

2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

Organs were not weighed for Groups 1 and 3 because all animals were found 

dead or humanely sacrificed. 
Huntingdon Life Sciences summary statistics for absolute organ weights (g) Printed: 06-Jan-03 

Princeton Research Center Study number: 025449 

East Millstone, New Jersey Version '4-·. 2." 

Terminal Sacrifice 

Animal Group/ Terminal Brain Liver Testes 

No/sex Subgroup Body wt. (g) Adrenal Glands Kidneys Lungs 

----------------------------------------------------------------------------------------------------------------------------------- M a 1 e A n i m a 1 s 

2046/M 2/1 377 .o 0.0684 2.2544 3.4068 17.6379 1.6755 3.2399 

2047/M 2/1 407.0 0,0600 2.0259 3.5122 17.3541 1. 9566 2.9476 

2048/M 2/1 388.0 0.0685 2.0765 3.4737 18.2689 1.8488 3.3805 

2049/M 2/1 366.0 0.0646 2.3692 3.5702 18.1211 1.7438 3.5192 

2050/M 2/1 329.0 0.0571 2.2754 3.5959 14.9046 1.7344 3.4621 

M e a n: 373.4 0.0637 2.2003 3. 5118 17.2573 1.7918 3.3099 

Standard deviation: 29.1 0.0051 0.1439 0.0758 1. 3656 0.1113 0.2281 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

4046/M 4/1 335.0 0.0576 1.8952 2.7983 14.3762 1.4783 3.1343 

4047/M 4/1 337.0 0.0563 1. 9204 3.0356 14.1729 1,4080 3.0534 

4048/M 4/1 341.0 0.0429 1.9985 3 .1143 15.4086 1.4637 2.9500 

4049/M 4/1 318.0 0.0620 1.8947 2.6141 13.7135 1.6069 3.0439 

4050/M 4/1 373.0 0.0667 1.8022 3.1793 14.8588 1. 3639 3.2847 

M e a n: 340.8 0.0571 1.9022 2.9483 14.5060 1. 4642 3.0933 

St.a.ndard deviation: 20.0 0.0089 0.0702 0.2360 0.6510 0.0919 0.1254 

Number of observ. ; (5) (5) (5) (5) (5) (5) (5) 

5046/M 5/1 367,0 0.0576 1.9707 3.1005 16.0453 1. 6682 2.9134 

5048/M 5/1 358.0 0.0464 2.0955 3.5442 17.1436 1.8335 3.1353 

5050/M 5/1 314.0 0.0457 1.9977 2.8845 12.7094 1. 2072 3.0363 

M e a n: 346.3 0.0499 2.0213 3.1764 15.2994 1,5696 3.0283 

Standard deviation: 28.4 0.0067 o·. 0657 0.3363 2.3093 0. 3246 0. 1112 

Number of observ. ; (3) (3) (3) (3) (3) (3) (3) 

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(0 

CD 

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Huntingdon Life Sciences Summary statistics for absolute organ weights (g) Printed: 06-Jan-03 

Princeton Research Center Study number: 025449 

East Millstone, New Jersey Vers"lon 4 . 2 . 2 

Terminal Sacrifice 

Animal Group/ Terminal Brain Liver Ovaries 

No/sex subgroup Body wt. (g) Adrenal Glands Kidneys Lungs 

----------------------------------------------------------------------------------------------------------------------------------- F e m a l e A n i m a l s 

2546/F 2/1 205.0 0. 0710 1. 9891 2 .1349 8. 8112 1.2743 0.0947 

2547/F 2/1 246.0 0.0680 2.0002 2.4986 10.9119 1. 4844 0.0915 

2548/F 2/1 235.0 0.0748 1.8890 2.3878 10.0819 1. 4285 0.0882 

2549/F 2/1 231.0 0.0781 1. 9120 2.5966 9.2695 1.4018 0.0904 

2550/F 2/1 256.0 0.0662 2.0361 2.2515 11.8970 1.4407. 0.1025 

M e a n: 234.6 0.0716 1.9653 2.3739 10.1943 1.4059 0.0935 

Standard deviation: 19.2 0.0049 0.0622 0.1854 1.2445 0.0794 0.0056 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

4546/F 4/1 280.0 0. 0722 1. 9596 2.5268 15.2965 1. 5157 0. 1125 

4547/F 4/1 232.0 0.0680 1.9099 2.0489 9.0264 1.2297 0.1093 

4548/F 4/1 247.0 0.0787 1.7682 2.0898 10.5843 1. 3648 0.1095 

4549/F 4/1 247.0 0.0824 1.9750 2.1749 10.1937 1. 2442 0.0754 

4550/F 4/1 248.0 0.0789 1.8657 2.0759 10.4412 1.7988 0.1033 

M e a n: 250.8 0.0760 1.8957 2.1833 11.1084 1.4306 0.1020 

Standard deviation: 17.6 0.0058 0.0833 0.1977 2.4203 0.2357 0.0152 

Number of observ. :. (5) (5) (5) (5) (5) (5) (5) 

5546/F 5/1 234.0 0.0744 1.8000 2.1160 10.8435 1.3010 0.1023 

5547/F 5/1 236.0 0. 0611 1.9149 1.8109 10.6102 1.2447 0.0787 

5548/F 5/1 238.0 0.0665 1. 9758 2.0299 10.7891 1.3141 0 .ll05 

5549/F 5/1 237.0 0.0610 1.9325 2.0676 10.9097 1.4109 0.1000 

5550/F 5/1 247.0 0.0743 1. 8833 2.0838 10.3378 l. 3334 0.0812 

M e a n: 238.4 0.0675 1.9013 2.0216 10.6981 1.3208 0.0945 

Standard deviation: 5.0 0.0067 0.0658 0.1218 0.2301 0.0602 0.0139 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

"'0 

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Huntingdon Life Sciences summary Statistics for % Organ to Body Weight Printed: 06-Jan-03 

Princeton Research Center Study number: 025449 

East Millstone, New Jersey Version 4.2.2 

Terminal Sacrifice 

Animal Group/ Terminal Brain Liver Testes 

No/sex Subgroup Body wt. (g) Adrenal Glands Kidneys Lungs 

----------------------------------------------------------------------------------------------------------------------~~-----------

M a l e A n i m a l s 

2046/M 2/1 377.0 0.0181 0.5980 0.9037 4.6785 0.4444 0.8594 

2047/M 2/1 407.0 0.0147 0.4978 0.8629 4.2639 0.4807 0. 7242 

2048/M 2/1 388.0 0.0177 0.5352 0.8953 4.7085 0.4765 0.8713 

2049/M 2/1 366.0 0.0177 0.6473 0.9755 4.9511 0.4764 0.9615 

2050/M 2/1 329.0 0.0174 0.6916 1.0930 4.5303 0. 5272 1. 0523 

M e a n: 373.4 0. 0171 0.5940 0.9461 4.6265 0.4811 0.8937 

Standard deviation: 29.1 0.0014 0.0792 0.0918 0.2528 0.0296 0.1226 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

4046/M 4/1 335.0 0. 0172 0.5657 0.8353 4.2914 0.4413 0.9356 

4047/M 4/1 337.0 0.0167 0.5699 0.9008 4.2056 0.4178 0.9061 

4048/M 4/1 341.0 0.0126 0.5861 0.9133 4.5187 0.4292 0.8651 

4049/M 4/1 318.0 0.0195 0.5958 0.8220 4.3124 0.5053 0. 9572 

4050/M 4/1 373.0 0.0179 0.4832 0.8524 3.9836 0.3657 0.8806 

M e a n: 340.8 0.0168 0,5601 0.8648 4.2623 0.4319 0.9089 

Standard deviation: 20.0 0.0026 0.0447 0.0403 0.1936 0.0502 0.0380 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

5046/M 5/1 367.0 0.0157 0.5370 0.8448 4.3720 0.4546 0.7938 

5048/M 5/1 358.0 0.0130 0.5853 0.9900 4.7887 0.5122 0.8758 

5050/M 5/1 314.0 0.0146 0.6362 0.9186 4.0476 0.3845 0.9670 

M e a n: 346.3 0.0144 0.5862 0.9178 4.4028 0.4504 0.8789 

Standard deviation: 28.4 0.0014 0.0496 0. 0726 0.3715 0.0639 0.0866 

Number of observ. : (3) (3) (3) (3) (3) (3) (3) 

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Huntingdon Life Sciences summary Statistics for % Organ to Body Weight Printed: 06-Jan-03 

Princeton Research Center Study number: 025449 

East Millstone, New Jersey Version 4.2 .. 2 

Terminal Sacrifice 

Animal Group/ Terminal Brain Liver Ovaries 

No/sex subgroup Body wt. (g) Adrenal Glands Kidneys Lungs 

------------------------------------------------------------------------------------------------------------------------~----------

Female A n i m a l s 

2546/F 2/1 205.0 0.0346 0.9703 1. 0414 4.2981 0.6216 0.0462 

2547/F 2/1 246.0 0.0276 0.8131 1·. 0157 4.4357 0.6034 0.0372 

2548/F 2/1 235.0 0.0318 0.8038 1. 0161 4.2902 0.6079 0.0375 

2549/F 2/1 231.0 0.0338 0.8277 1.1241 4.0128 0.6068 0.0391 

2550/F 2/1 256.0 0.0259 0.7954 0.8795 4.6473 0.5628 0.0400 

M e a n: 234.6 0.0308 0.8421 1.0154 4.3368 0.6005 0.0400 

Standard deviation: 19.2 0.0038 0.0727 0.0880 0.2318 0.0222 0.0036 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

4546/F 4/1 280.0 0.0258 0.6999 0.9024 5.4630 0.5413 0.0402 

4547/F 4/1 232.0 0.0293 0.8232 0.8831 3.8907 0.5300 0.0471 

4548/F 4/1 247.0 0.0319 0. 7159 0.8461 4.2851 0.5526 0.0443 

4549/F 4/1 247.0 0.0334 0.7996 0.8805 4.1270 0.5037 0.0305 

4550/F 4/1 248.0 0.0318 0.7523 0.9371 4.2102 o. 7253 0.0417 

M e a n: 250.8 0.0304 0.7582 0.8698 4.3952 0.5706 0.0409 

Standard deviation: 17.6 0.0030 0.0529 0.0274 0.6150 0.0884 0.0063 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

5546/F 5/1 234.0 0.0318 0.7692 0.9043 4.6340 0.5560 0.0437 

5547/F 5/1 236.0 0.0259 0. 8114 0.7673 4.4958 0.5274 0.0333 

5548/F 5/1 239.0 0.0279 0.9302 0.8529 4.5332 0.5521 0.0464 

5549/F 5/1 237.0 0.0257 0.9154 0. 8724 4.6032 0.5953 0.0422 

5550/F 5/1 247.0 0.0301 0.7625 0.9436 4.1953 0.5399 0.0329 

M e a n: 238.4 0.0283 0.7977 0.8481 4.4903 0.5541 0.0397 

Standard deviation: 5.0 0.0026 0.0300 0.0508 0.1791 0.0256 0.0062 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

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Huntingdon Life Sciences Summary Statistics for % Organ. to Brain Weight ?rinted: 06-Jan-03 

?rinceton Research Center Study number: 025449 

East Millstone, New Jersey Version 4.2.2 

Terminal sacrifice 

Animal Group/ Terminal Brain Liver Testes 

No/sex Subgroup Body wt. (g) Adrenal Glands Kidneys Lungs 

----------------------------------------------------------------------------------------------------------------------------------- M a 1 e A n i m a 1 s 

2046/M 2/1 377.0 3.0341 100.0000 151.1178 782.3767 74.3213 143.7145 

2047/M 2/1 407.0 2.9616 100.0000 173,3649 856.6119 96.5793 145.4958 

2048/M 2/1 388.0 3.2988 100.0000 167.2863 879.7930 89.0344 162.7980 

2049/M 2/1 366.0 2. 7267 100.0000 150.6922 764.8616 73.6029 148.5396 

2050/M 2/1 329.0 2.5094 100.0000 158.0338 655.0322 76.2240 152.1535 

M e a n: 373.4 2.9061 100.0000 160.0990 787.7351 81.9524 150.5403 

Standard deviation: 29.1 0.3013 0.0000 10.0132 88.5729 10.3061 7.5646 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

4046/M 4/1 335.0 3.0393 100.0000 147.6520 758.5585 78.0023 165.3810 

4047/M 4/1 337,0 2.9317 100.0000 158.0712 738.0182 73.3181 158.9981 

4048/M 4/1 341.0 2.1466 100.0000 155.8319 771.0083 73.2399 147.6107 

4049/M 4/1 318.0 3.2723 100.0000 137.9691 723.7822 84 '8103 160.6534 

4050/M 4/1 373.0 3.7010 100.0000 176.4122 824.4813 75.6797 182.2606 

M e a n: 340.8 3.0182 100.0000 155 .. 1873 763.1697 77.0101 162.9808 

Standard deviation: 20.0 0.5697 0.0000 14.2520 38.8130 4.7813 12.5995 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

5046/M 5/1 367.0 2.9228 100.0000 157.3299 814.1930 84.6501 147.8358 

5048/M 5/1 358.0 2.2143 100.0000 169.1339 818.1151 87.4970 149.6206 

5050/M 5/1 314.0 2.2876 100.0000 144.3911 636.2017 60.4295 151.9898 

M e a n: 346.3 2.4749 100.0000 156.9516 756.1699 77.5256 149.8154 

Standard deviation: 28.4 0.3896 0.0000 12.3757 103.9140 14.8739 2.0838 

Number of observ. : (3) (3) (3) (3) (3) (3) (3) 

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Huntingdon Life Sciences Summary Statistics for % Organ to Brain Weight Printed: 06-Jan-03 

Princeton Research Center study number: 025449 

East Millstone, New Jersey Version 4.2.2 

Terminal Sacrifice 

Animal Group/ Terminal Brain Liver Ovaries 

No/sex Subgroup Body wt. (g) Adrenal Glands Kidneys Lungs 

----------------------------------------------------------------------------------------------------------------------.------------- ·Female A n i m a 1 s 

2546/F 2/1 205.0 3.5695 100.0000 107.3300 442.9743 64.0642 4.7609 

2547/F 2/1 246.0 3.3997 100.0000 124.9175 545.5405 74.2126 4.5745 

2548/F 2/1 235.0 3.9598 100.0000 126.4055 533.7163 75.6220 4.6691 

2549/F 2/1 231.0 4.0847 100.0000 135.8055 484.8065 73.3159 4.7280 

2550/F 2/1 256.0 3.2513 100.0000 110.5791 584.3034. 70.7578 5.0341 

M e a n: 234.6 3.6530 100.0000 121.0075 518.2682 71.5945 4.7534 

Standard deviation: 19.2 0.3581 0.0000 11.8241 55.0725 4.5674 0.1722 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

4546/F 4/1 280.0 3.6844 100.0000 128.9447 780.5931 77.3474 5.7410 

4547/F 4/1 232.0 3.5604 100.0000 107.2779 472.6112 64.3856 5. 7228 

4548/F 4/1 247.0 4.4509 100.0000 118.1880 598.5919 77.1858 6.1927 

4549/F 4/1 247.0 4.1722 100.0000 110.1215 516.1368 62.9975 3.8177 

4550/F 4/1 248.0 4.2290 100.0000 111.2666 559.6399 96.4142 5.5368 

M e a: n: 250.8 4.0194 100.0000 115.1597 585.5146 75.6661 5.4022 

Standard deviation: 17.6 0.3796 0.0000 8.6870 118.8030 13.4477 0.9180 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

· 5546/F 5/1 234.0 4.1333 100.0000 117.5556 602.4167 72.2778 5,6833 

5547/F 5/1 236.0 3.1908 100.0000 94.5669 554.0665 65.0008 4.1099 

5548/F 5/1 238.0 3.3657 100.0000 102.7381 546.0624 66.5098 5.5927 

5549/F 5/1 237.0 3.1565 100.0000 106.9910 564.5383 73.0091 5.1746 

5550/F 5/1 247.0 3.9452 100.0000 110.6462 548.9196 70.8013 4. 3116 

M e a n: 238.4 3.5583 100.0000 106.5000 563.2047 69.5197 4.9744 

Standard deviation: 5.0 0.4511 0.0000 8·.6047 23.0241 3.5674 0.7266 

Number of observ. : (5) (5) (5) (5) (5) (5) (5) 

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Huntingdon Life Sciences 02-5449 Page 76 

Final Report 

Incidence Summary Report 

', for Gross Necropsy ObseNations 

Preface Table 9 

Table of Contents 

Terminal Sacrifice ............................................................................................... 77 

Unscheduled Deaths ............................................................................................ 78 

All Sacrifices ....................................................................................................... 79 

Corresponding exposure levels for each group were as follows: 

Group 1 

Group 2 

Group 3 

Group 4 

Group 5 

2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene (24,800 ppm) 

2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 
Huntingdon Life Sciences 

Princeton Research Center 

East Millstone, New Jersey 

Incidence Summary Report for Gross Necropsy Observations 

Study number: 025449 

Printed: 02-Jan-03 

Page: 1 

Version 4.2.2 

Terminal Sacrif=i~c~e----------------------------------~------------------~---------------------------------------------------------------- - Males ·- I -- Females 

Group: 

Number in group: 

Within normal limits ............... 

Lungs 

Discolored ..................... Adhesion ....................... 

1 

0 

0 

0 

0 

2 3 4 5 1 1 2 3 

5 o s 3 I o s o 

4 0 5 3 0 4 0 

1 0 0 0 0 0 0 

0 0 0 0 0 1 0 

4 

5 

4 

1 

0 

5 

5 

5 

0 

0 

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CD 

"" "" 
Huntingdon Life Sciences 

Princeton Research Center 

East Millstone, New Jersey 

Unscheduled Deaths 

Group: 

Number in group: 

Within normal limits •• 0 •• 0 .... 0. 0. 0 

Lungs 

Discolored ..................... 

Abnormal Contents ...... 0 •••• 0 ••• 

Trachea 

Abnormal Contents 0. 0 .... 0 0 ••• 0 .. 0 

1 

5 

2 

3 

1 

1 

Incidence summary Report for Gross Necropsy Observations 

study number: 025449 

Males 

2 3 

0 5 

0 0 

0 5 

0 0 

0 0 

4 

0 

0 

0 

0 

0 

5 

2 

0 

0 

2 

0 

I 

I 

I 

1 

5 

3 

2 

0 

0 

-- Females 

2 3 

0 5 

0 0 

0 5 

0 0 

0 0 

4 

0 

0 

0 

0 

0 

5 

0 

0 

0 

0 

0 

Printed: 02-Jan-03 

Page: 1 

Version 4.2.2 

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(X) 
Huntingdon Life Sciences 

Princeton Research Center 

East Millstone, New Jersey 

All Sacrifices 

Group: 

Number in group: 

Within normal limits ......... ~ .... 

Lungs 

Discolored ..................... 

Adhesion ....................... 

Abnormal Contents .............. 

Trachea 

Abnormal Contents .............. 

1 

5 

2 

3 

0 

1 

1 

Incidence Summary Report for Gross Necropsy Observations 

Study number: 025449 

Males 

2 3 

5 5 

4 0 

1 5 

0 0 

0 0 

0 0 

4 

5 

5 

0 

0 

0 

0 

5 

5 

3 

0 

0 

2 

0 

I 

I 

I 

1 

5 

3 

2 

0 

0 

0 

-- Females 

2 3 

5 5 

4 0 

0 5 

1 0 

0 0 

0 0 

4 

5 

4 

1 

0 

0 

0 

5 

5 

5 

0 

0 

0 

0 

Printed: 02-Jan-03 

Page: 1 

Version 4.2.2 

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Huntingdon Life Sciences 

', 

Table of Contents 

02-5449 

Lesion Incidence Summary 

with Expanded Severity Levels 

Preface 

Page 80 

Final Report 

Table 10 

Terminal Sacrifice ............................................................................................... 81 

Unscheduled Deaths ............................................................................................ 83 

All Sacrifices ....................................................................................................... 86 

Key to Abbreviations 

NAD = No abnormal diagnoses 

Corresponding exposure levels for each group were as follows: 

Group 1 

Group 2 

Group 3 

Group 4 

Group 5 

Note: 

2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene (24,800 ppm) 

2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

Due to computer limitations, Group 1 appears as CTLS (Controls) on the 

following table. There was no Control Group for this study. 
02-Jan-03; 18:28 

Huntingdon Life Sciences 025449 

-

An Acute (4-Hour) Inhalation Toxicity study 

in the Rat via Nose-Only Exposure 

Lesion Incidence Summary With Expanded Severity Levels __________________________________ ; __ ! __________________________________________________________ ·--~~-------------------------------

-- A n i m a l s A f f e c t e d 

Controls from group(s): 1 Animal sex: -- M a l e s -- I -- F e m a l e s 

Dosage group: Ctls 2 3 4 5 I Ctls 2 3 4 5 

T i s s u e s w i t h D i a 9 n o s e s No. in grouE: 0 5 0 5 3 I 0 5 0 5 5 

Lungs ..................•....................... Number examined: 0 5 0 5 3 I 0 5 0 5 5 

CONGESTION 

Nad> 0 5 0 5 3 I 0 5 0 4 5 

Slight> 0 0 0 0 0 I 0 0 0 1 0 

............................ Total Incidence of Finding Observed: 0 0 0 0 0 I 0 0 0 1 0 

EDEMA 

Nad> 0 5 0 5 3 I 0 5 0 5 5 

............................ Total Incidence of Finding Observed: 0 0 0 0 0 I 0 0 0 0 0 

HEMORRHAGE(£) 

Nad> 0 4 0 5 3 I 0 4 0 4 5 

Slight> 0 1 0 0 0 I 0 1 0 1 0 

..................•......... Total Incidence of Finding Observed: 0 l 0 0 0 I 0 1 0 1 0 

ARTERIES: MEDIAL HYPERTROPHY 

Nad> 0 3 0 1 0 I 0 2 0 2 3 

Minimal> 0 2 0 4 3 I 0 3 0 3 2 

............................ Total Incidence of Finding Observed: 0 2 0 4 3 I 0 3 0 3 2 

ALVEOLAR/INTRAALVEOLAR MACROPHAGE£ 

Minimal> 0 2 0 3 3 I 0 l 0 3 4 

Slight> 0 3 0 2 0 I 0 4 0 2 1 

............................ Total Incidence of Finding Observed: 0 5 0 5 3 I 0 5 0 5 5 

BRONCHIOLAR EPITHELIUM: DEQUAMATED 

Nad> 0 5 0 5 3 I 0 5 0 5 5 

............................ Total Incidence of Finding Observed: 0 0 0 0 0 I 0 0 0 0 0 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE INFLAMMATION 

Nad> 0 5 0 5 3 I 0 5 0 5 5 

..... , ...................... Total Incidence of Finding Obs.erved: 0 0 0 0 0 I 0 0 0 0 0 

All Diagnoses; Death types: All scheduled; Date of death range: 01-0ct-02 To 04-Nov-02 

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_... 
02-Jan-03; 18:28 

Huntingdon Life Sciences 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Lesion Incidence summary With Expanded Severity Levels 

-- A n i m a 1 s 

Controls from group(s): 1 Animal sex: -- M a 1 e s --

Dosage group: Ctls 2 3 4 5 

T i s s u e s w i t h D i a s n o s e s No. in grouE: 0 5 0 5 3 

Lungs .......................................... Number examined: 0 5 0 5 3 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

-CELLS/INFLAMMATORY CELLS/CELL DEBRIS 

Nad> 0 5 0 5 3 

............................ Total Incidence of Finding Observed: 0 0 0 0 0 

LYMPHOID CELL AGGREGATE(S) 

Nad> 0 3 0 3 1 

Minimal> 0 2 0 2 2 

......................•..... Total Incidence of Finding Observed: 0 2 0 2 2 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC INFLAMMATION 

Nad> 0 2 0 5 3 

Minimal> 0 3 0 0 0 

Slight> 0 0 0 0 0 

............................ Total Incidence of Finding Observed: 0 3 0 0 0 

INTERSTITIUM: OSSEOUS METAPLASIA 

Nad> 0 5 0 4 2 

Minimal> 0 0 0 1 1 

............................ Total Incidence of Finding Observed: 0 0 0 1 1 

VESSEL(S): MINERAL DEPOSIT(S) 

Nad> 0 4 0 5 3 

Minimal> 0 1 0 0 0 

............................ Total Incidence of Finding Observed: 0 1 0 0 0 

VISCERAL PLEURA/CAPSULE: FIBROSIS 

Nad> 0 5 0 5 3 

Minimal> 0 0 0 0 0 

.................•.......... Total Incidence of Finding Observed: 0 0 0 0 0 

All Diagnoses; Death types: All scheduled; Date of death range: 01-0ct-02 To 04-Nov-02 

A f f e c t e d 

I -- F e m a 1 e s 

I Ctls 2 3 4 

I 0 5 0 5 

I 0 5 0 5 

I 0 5 0 5 

I 0 0 0 0 

I 0 1 0 1 

I 0 4 0 4 

I 0 4 0 4 

0 4 0 5 

0 0 0 0 

0 1 0 0 

0 1 0 0 

I 0 5 0 5 

I 0 0 0 0 

I 0 0 0 0 

I 0 5 0 5 

I 0 0 0 0 

I 0 0 0 0 

I 0 4 0 5 

I 0 1 0 0 

I 0 1 0 0 

5 

5 

5 

5 

0 

4 

1 

1 

5 

0 

0 

0 

4 

1 

1 

5 

0 

0 

5 

0 

0 

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02-Jan-03; 18:28 

Huntingdon Life Sciences 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Lesion Incidence Summary With Expanded Severity Levels 

-- An i m a 1 s 

Controls from group(s): 1 Animal sex: -- M a 1 e s --

Dosage group: Ctls 2 3 4 5 

T i s s u e s w i t h D i a g n o s e s No. in grou12: 5 0 5 0 2 

Lungs .......•.................................. Number examined : 5 0 5 0 2 

CONGESTION 

Nad> 3 0 0 0 l 

Minimal> 0 0 0 0 0 

Slight> l 0 3 0 0 

Moderate> 1 0 2 0 1 

............................ Total Incidence of Finding Observed: 2 0 5 0 l 

EDEMA 

Nad> 4 0 5 0 2 

Slight> 1 0 0 0 0 

.......................••... Total Incidence of Finding Observed: l 0 0 0 0 

HEMORRHAGE(S) 

Nad> 3 0 4 0 2 

Minimal> 0 0 0 0 0 

Slight> 2 0 1 0 0 

............................ Total Incidence of Finding Observed: 2 0 1 0 0 

ARTERIES: MEDIAL HYPERTROPHY 

Nad> 0 0 1 0 2 

Minimal> 4 0 3 0 0 

Slight> 1 0 1 0 0 

............................ Total Incidence of Finding Observed: 5 0 4 0 0 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES 

Minimal> 0 0 2 0 2 

Slight> 2 0 3 0 0 

Moderate> 3 0 0 0 0 

............................ Total Incidence of Finding Observed: 5 0 5 0 2 

All Diagnoses; Death types: All unscheduled; Date of death range: 17-Sep-02 To 21-0ct-02 

A f f e c t e d 

I -- F e m a 1 e s 

I Ctls 2 3 4 

I 5 0 5 0 

I 5 0 5 0 

l 0 0 0 

l 0 ·0 0 

3 0 4 0 

0 0 1 0 

4 0 5 0 

I 5 0 5 0 

I 0 0 0 0 

I 0 0 0 0 

4 0 4 0 

0 0 1 0 

1 0 0 0 

1 0 1 0 

1 0 2 0 

4 0 3 0 

0 0 0 0 

4 0 3 0 

0 0 3 0 

4 0 2 0 

1 0 0 0 

5 0 5 0 

,j. 

5 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

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02-Jan-03; 18:28 

Huntingoon Life Sciences 025449 

--, 

An Acute (4-Hour) Inhalation Toxicity Stuoy 

in the Rat via Nose-Only Exposure 

Lesion Incioence Summary With Expanded Severity Levels 

-- A n i m a 1 s 

Controls from group(s): l Animal sex: -- M a 1 e s --

Dosage group: Ctls 2 3 4 5 I 

T i s s u e s W i t h D i a g n o s e s No. in group: 5 o 5 o 2 I 

Lungs ........................•.....•........... Number examined: 5 o 5 0 2 I 

BRONCHIOLAR EPITHELIUM: DEQUAMATED 

Nad> 

Slight> 

Moderate> 

............................ Total Incidence of Finding Observed: 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE INFLAMMATION 

Nad> 

Slight> 

.........................•.. Total Incidence of Finding Observed: 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

-CELLS/INFLAMMATORY CELLS/CELL DEBRIS 

Nad> 

Slight> 

Moderate> 

............................ Total Incidence ctf Finding Observed: 

LYMPHOID CELL AGGREGATE(S) 

Nad> 

Minimal> 

................... , ........ Total Incidence of Finding Observed: 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC INFLAMMATION 

Nad> 

Minimal> 

............................ Total Incidence of Finding Observed: 

INTERSTITIUM: OSSEOUS METAPLASIA 

Nad> 

Minimal> 

.....................••..... Total Incidence of Finding Observed: 

0 

l 

4 

5 

0 

5 

5 

2 

0 

3 

3 

4 

l 

l 

2 

3 

3 

5 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

2 

3 

3 

5 

0 

0 

4 

l 

l 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

All Diagnoses; Death types: All unscheduled; Date of death range: 17-Sep-02 To 21-0ct-02 

2 

0 

0 

0 

2 

0 

0 

2 

0 

0 

0 

l 

l 

l 

2 

0 

0 

2 

0 

0 

A f f e c t e d 

F e m a 1 e s 

ctls 2 3 4 

5 

5 

0 

4 

l 

5 

0 

5 

5 

0 

5 

0 

5 

5 

0 

0 

3 

2 

2 

4 

l 

l 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

5 

5 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

2 

3 

3 

5 

0 

0 

5 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

5 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

0 

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02-Jan-03; 18:28 

Huntingdon Life Sciences 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Lesion Incidence summary With Expanded Severity Levels 

-- A n i m a 1 s 

Controls from group(s): 1 Animal sex: -- M a 1 e s --

Dosage group: Ctls 2 3 4 5 I 

T i s s u e s W i t h D i a g n o s e s No. in group: 5 0 5 o 2 _ _j 

Lungs ...............•.......................... Number examined: 5 o . 5 o 2 I 

VESSEL(S): MINERAL DEPOSIT(S) 

Nad> 5 0 5 0 2 I ............................ Total Incidence of Finding Observed: 0 o o o o I 

VISCERAL PLEURA/CAPSULE: FIBROSIS 

Nad> 

............................ Total Incidence of Finding Observed: 

5 

0 

0 

0 

5 

0 

0 

0 

All Diagnoses; Death types: All unscheduled; Date of death range: 17-Sep-02 To 21-0ct-02 

2 

0 

A f f e c t e d 

F e m a 1 e s 

Ctls 2 3 4 

5 0 5 0 

5 0 5 0 

5 0 5 0 

0 0 0 0 

5 0 5 0 

0 0 0 0 

5 

0 

0 

0 

0 

0 

0 

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02-Jan-03; 18:29 

Huntingdon Life Sciences 025449 

An Acute (4-Hour) Inhalation.Toxicity Study 

in the Rat via Nose-Only Exposure 

Lesion Incidence Summary With Expanded Severity Levels 

-- A n i m a l s 

Controls from group(s): 1 Animal sex: -- M a 1 e s --

Dosage group: Ctls 2 3 4 5 

T i s s u e s w i t h D i a g n o s e s No. in grou12: 5 5 5 5 5 

Lungs .......................................... Number examined: 5 5 5 5 5 

CONGESTION 

Nad> 3 5 0 5 4 

Minimal> 0 0 0 0 0 

Slight> 1 0 3 0 0 

Moderate> 1 0 2 0 1 

..•......................... Total Incidence of Finding observed: 2 0 5 0 1 

EDEMA 

Nad> 4 5 5 5 5 

Slight> 1 0 0 0 0 

......................•..... Total Incidence of Finding Observed: 1 0 0 0 0 

HEMORRHAGE(S) 

Nad> 3 4 4 5 5 

Minimal> 0 0 0 0 0 

Slight> 2 1 1 0 0 

.................•.......... Total Incidence of Finding Observed; 2 1 1 0 0 

ARTERIES: MEDIAL HYPERTROPHY 

Nad> 0 3 1 1 2 

Minimal> 4 2 3 4 3 

Slight> 1 0 1 0 0 

...................•........ Total Incidence of Finding Observed: 5 2 4 4 3 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES 

Minimal> 0 2 2 3 5 

Slight> 2 3 3 2 0 

Moderate> 3 0 0 0 0 

.........................••. Total Incidence of Finding Observed: 5 5 5 5 5 

All Diagnoses; Death types: All; Date of death range: 17-Sep-02 To 04-Nov-02 

A f f e c t e d 

I -- F e m a l e s 

I Ctls 2 3 4 

I 5 5 5 5 

I 5 5 5 5 

1 5. 0 4 

1 0 0 0 

3 0 4 1 

0 0 1 0 

4 0 5 1 

I 5 5 5 5 

I 0 0 0 0 

I 0 0 0 0 

4 4 4 4 

0 0 1 0 

1 1 0 1 

1 1 1 1 

I 1 2 2 2 

I 4 3 3 3 

I 0 0 0 0 

I 4 3 3 3 

I 0 1 3 3 

I 4 4 2 2 

I 1 0 0 0 

I 5 5 5 5 

5 

5 

5 

5 

0 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

3 

2 

0 

2 

4 

1 

0 

5 

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02-Jan-03; 18:29 

Huntingdon Life Sciences 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Lesion Incidence Summary With Expanded Severity Levels 

-- An i m a 1 s 

Controls from group(s): 1 Animal sex: -- M a l e s -- I 

Dosage group: Ctls 2 3 4 5 I 

T i s s u e s W i t h D i a g n o s e s No. in group: 5 5 5 5 5 I 

Lungs .......................................... Number examined: 5 5 5 5 5 I 

BRONCHIOLAR EPITHELIUM: DEQUAMATED 

Nad> 

Slight> 

Moderate> 

............................ Total Incidence of Finding Observed: 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE INFLAMMATION 

Nad> 

Slight> 

............................ Total Incidence of Finding Observed: 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

-CELLS/INFLAMMATORY CELLS/CELL DEBRIS 

Nad> 

Slight> 

Moderate> 

............................ Total Incidence of Finding Observed: 

LYMPHOID CELL AGGREGATE(S) 

Nad> 

Minimal> 

............................ Total Incidence of Finding Observed: 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC INFLAMMATION 

Nad> 

Minimal> 

Slight> 

... , . · .................••.... Total Incidence of Finding Observed: 

INTERSTITIUM: OSSEOUS METAPLASIA 

Nad> 

Minimal> 

...............•............ Total Incidence of Finding Observed: 

0 

1 

4 

5 

0 

5 

5 

2 

0 

3 

3 

4 

1 

1 

2 

3 

0 

3 

5 

0 

0 

All Diagnoses; Death types: All; Date of death range: 17-Sep-02 To 04-Nov-02 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

3. 

2 

2 

2 

3 

0 

3 

5 

0 

0 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

2 

3 

3 

5 

0 

0 

0 

4 

1 

1 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

3 

2 

2 

5 

0 

0 

0 

4 

1 

1 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

2 

3 

3 

5 

0 

0 

0 

4 

1 

1 

A f f e c t e d 

F e m a l 

Ctls 

5 

5 

0 

4 

1 

5 

0 

5 

5 

0 

5 

0 

5 

5 

0 

0 

3 

2 

0 

2 

4 

1 

1 

2 3 

5 5 

5 5 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

1 

4 

4 

4 

0 

1 

1 

5 

0 

0 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

2 

3 

3 

.5 

0 

0 

0 

5 

0 

0 

e s 

4 

5 

5 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

1 

4 

4 

5 

0 

0 

0 

5 

0 

0 

5 

5 

5 

5 

0 

0 

0 

5 

0 

0 

5 

0 

0 

0 

4 

1 

1 

5 

0 

0 

0 

4 

1 

1 

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02-Jan-03; 18:29 

Huntingdon Life Sciences 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Lesion Incidence summary With Expanded Severity Levels 

-- A n i m a 1 s 

Controls from group(s): 1 Animal sex: -- M a 1 e s --

Dosage group: Ctls 2 3 4 5 

T i s s u e s w i t h D i a q n o s e s No. in s:rou12: 5 5 5 5 5 

Lungs .......................................... Number examined: 5 5 5 5 5 

VESSEL(S): MINERAL DEPOSIT(S) 

Nad> 5 4 5 5 5 

Minimal> 0 1 0 0 0 

..................•......... Total Incidence of Finding Observed: 0 1· 0 0 0 

VISCERAL PLEURA/CAPSULE: FIBROSIS 

Nad> 5 5 5 5 5 

Minimal> 0 0 0 0 0 

............................ Total Incidence of Finding Observed: 0 0 0 0 0 

All Diagnoses; Death types: All; Date of death range: 17-Sep-02 To 04-Nov-02 

A f f e c t e d 

I -- F e m a 1 e s 

I Ctls 2 3 4 

I 5 5 5 5 

I 5 5 5 5 

I 5 5 5 5 

I 0 0 0 0 

I 0 0 0 0 

I 5 4 5 5 

I 0 1 0 0 

I 0 1 0 0 

5 

5 

5 

5 

0 

0 

5 

0 

0 

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Huntingdon Life Sciences 02-5449 Page 89 

Final Report 

Individual Animal Gross 

and Microscopic Observations 

', Preface Table 11 

Corresponding exposure levels for each group were as follows: 

Group 1 

Group 2 

Group 3 

Group 4 

Group 5 

Note: 

2-Bromo-3,3,3-Trifluoropropene (26,580 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (25,650 ppm) 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene (24,800 ppm) 

2-Bromo-3,3,3-Trifluoropropene (5, 173 ppm) 

E-1-Bromo-3,3,3-Trifluoropropene (45,850 ppm) 

Unless otherwise specified in a histopathology note, the organ/tissue examined 

was the required (routine) section. 
02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 1 

STATUS: Humane/Otherwise Specified 

PHASE DAY OF DEATH: 1 

SEX: Male 

PHASE: Dosing phase 

ANIMAL: _1046 

GROUP: 1 

Tissue Gross Observations/Comments 

Lungs . ....... No gross observations on tissue. 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, 

Moderate. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC 

INFLAMMATION, Multifocal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 2 

STATUS: Humane/Otherwise Specified 

PHASE DAY OF DEATH: l 

SEX: Male 

PHASE: Dosing phase 

ANIMAL: 1047 

GROUP·; 1 

Tissue 

Lungs . · · · · · · · · 

Gross Observations/Comments 

Discolored, All lobes, Red, Foci, 0.2 - 0.5 

em, Moderate 

Microscopic Observations/Comments 

Examined; 1 correlation found: 

HEMORRHAGE(S), Multifocal, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, 

Moderate. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

CELLS/INFLAMMATORY CELLS/CELL DEBRIS, 

Moderate. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC 

INFLAMMATION, Multifocal, Minimal. 

Tissues without comment under Gross observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan~03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 3 

STATUS: Humane/Otherwise Specified 

PHASE DAY OF DEATH: 1 

SEX: Male 

PHASE: Dosing phase 

ANIMAL(; 1048 

GROUP: 1 

Tissue 

Lungs ........ . 

Gross Observations/Comments 

Discolored, All lobes, Red, Moderate 

Discolored, Right lobes, Red, Foci, 0.2 -

0.5 em, Moderate 

Microscopic Observations/Comments 

Examined; 1 correlation found: 

CONGESTION, Slight. 

Examined; 1 correlation found: 

HEMORRHAGE(S), Multifocal, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Slight. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Moderate. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, 

Moderate. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

CELLS/INFLAMMATORY CELLS/CELL DEBRIS, 

Moderate. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 4 

STATUS: Humane/Otherwise Specified 

PHASE DAY OF DEATH: 1 

SEX: Male 

PHASE: Dosing phase 

ANIMAL: _1049 

GROUP: 1 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Moderate. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, Slight. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC 

INFLAMMATION, Multifocal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 5 

STATUS: .Found Dead 

PHASE DAY OF DEATH: 1 

SEX: Male 

PHASE: Dosing phase 

ANIMAL;. 1050 

GROUP: 1 

Tissue 

Lungs . · · · · · · · · 

Trachea . . . . . . . . 

Gross Observations/Comments 

Discolored, All lobes, Red, Severe 

Abnormal Contents, All lobes, Fluid, 

Moderate 

Abnormal Contents, Lumen, Clear, Frothy 

fluid, Moderate 

Microscopic Observations/Comments 

Examined; 1 correlation found: 

CONGESTION, Moderate. 

Examined; 1 correlation found: 

EDEMA, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Moderate. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, 

Moderate. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

CELLS/INFLAMMATORY CELLS/CELL DEBRIS, 

Moderate. 

No micropathology observations on tissue. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acu~e (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs •........ No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

Page: 6 

ANIMAL: .2046 

GROUP: 2 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC 

INFLAMMATION, Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue 

Lungs ........ . 

Gross Observations/Comments 

Discolored, All lobes, Red, Foci, 0.2 - 0.5 

em, Slight 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

Examined; 1 correlation found: 

HEMORRHAGE(S), Multifocal, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 7 

ANIMAL:,, 2047 

GROUP: 2 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

VESSEL(S): MINERAL DEPOSIT(S), Focal, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

Page: 8 

ANIMAL: "2048 

GROUP: 2 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC 

INFLAMMATION, Multifocal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 9 

--------~---------------------------------------------;------------------------------~---------------------------------------------

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

ANIMAL: 20.49 

GROUP: 2 

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02-Jan-03; 16:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs .....•... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC 

INFLAMMATION, Multifocal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 10 

ANIMAL:.,2050 

GROUP: 2 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH: 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Male 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs . . . . . . . . . 

Discolored, All lobes, Red, Slight Examined; l correlation found.: 

CONGESTION, Slight. 

Page: 11 

ANIMAL.: 3046 

GROUP: 3 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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Ol 

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CD 

...... 

0 

0 
02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH: 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

·Individual Animal Gross and Microscopic Observations 

SEX: Male 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs . . . . . . . · · 

Discolored, All lobes, Red, Moderate Examined; 1 correlation found: 

CONGESTION, Moderate. 

HEMORRHAGE(S), Multifocal, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Slight. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 12 

ANIMAL·:. 3 04 7 

GROUP: 3 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

INTERSTITIUM: OSSEOUS METAPLASIA, Focal, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH; 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Male 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs . · · · · · · · · Discolored, Right lobes, Red, Moderate Examined; 1 correlation found: 

CONGESTION, Moderate. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 13 

ANIMAL: .3048 

GROUP: 3 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH; 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Male 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs ........ . 

Discolored, All lobes, Red, Slight Examined; 1 correlation found: 

CONGESTION, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 14 

ANIMAL:, 3049 

GROUP: 3 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH: 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Male 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs . . · · · · · · · 

Discolored, All lobes, Red, Slight Examined; 1 correlation found: 

CONGESTION, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 15 

ANIMAL: .. 3050 

GROUP: 3 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE .DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs . . No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 16 

ANIMAL:. 4046 

GROUP: 4 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 17 

ANIMAL_: 4 04 7 

GROUP: 4 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

INTERSTITIUM: OSSEOUS METAPLASIA, Focal, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS:· Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 18 

ANIMAL: 4048 

GROUP: 4 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ...•..... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

LYMPHOID CELL AGGREGATE(S), Multifocal, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 19 

ANIMAL:_, 4049 

GROUP: 4 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) InhalaUon Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 20 

ANIMAL.: 4050 

GROUP: 4 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 21 

ANIMAL; 5046 

GROUP: 5 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

INTERSTITIUM: OSSEOUS METAPLASIA, 

Multifocal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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0 
02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH: 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic observations 

SEX: Male 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs ..... , ... 

Abnormal.Contents, All lobes, Fluid 

CONGESTION, Moderate. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 22 

ANIMA~,: 5047 

GROUP: 5 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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...... 
02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 23 

ANIMA~: 5048 

GROUP: 5 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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....... 

1\) 
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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH: 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Male 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs ........ . 

Abnormal Contents, All lobes, Fluid Examined; no correlation found 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 24 

ANIMAL: 5Q49 

GROUP: 5 

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P> 

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c.v 
02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Male 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 25 

ANIMAL: 5050 

GROUP: 5 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 26 

STATUS: Humane/Otherwise Specified 

PHASE DAY OF DEATH: 1 

SEX: Female 

PHASE: Dosing phase 

ANIMAL:. 1546 

GROUP: 1 

Tissue Gross Observations/Comments 

Lungs ....•.... No gross observations on tissue. 

Microscopic Observations/Comments 

HEMORRHAGE(S), Multifocal, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, Slight. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

BRONCHIOLAR LUMENS; DESQUAMATED EPITHELIAL 

CELLS/INFLAMMATORY CELLS/CELL DEBRIS, 

Slight. 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC 

INFLAMMATION, Multifocal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

~', 

An Acute (4-Hour) Inhalation Toxicity study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 27 

STATUS: Humane/Otherwise Specified 

PHASE DAY OF DEATH: 1 

SEX: Female 

PHASE: Dosing phase 

ANIMA!;; 1547 

GROUP: 1 

Tissue Gross Observations/Comments 

Lungs ........• No gross observations on tissue. 

Microscopic Observations/Comments 

CONGESTION, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, 

Moderate. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

CELLS/INFLAMMATORY CELLS/CELL DEBRIS, 

Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 28 

STATUS: Found Dead 

PHASE DAY OF DEATH: 1 

SEX: Female 

PHASE: Dosing phase 

ANIMAL~ 1548 

GROUP: l 

Tissue G'ross Observations/ Comments 

Lungs ........ . 

Discolored, All lobes, R~d, Moderate 

Microscopic Observations/Comments 

Examined; 1 correlation found: 

CONGESTION, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, Slight. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

CELLS/INFLAMMATORY CELLS/CELL DEBRIS, 

Slight. 

INTERSTITIUM: OSSEOUS METAPLASIA, Focal, 

Minimal. 

Tissues without ·comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

Page: 29 

STATUS: Humane/Otherwise Specified 

PHASE DAY OF DEATH: 1 

SEX: Female 

PHASE: Dosing phase 

ANIMAL,: 1549 

GROUP: 1 

Tissue Gross Observations/Comments 

Lungs ........ . 

Discolored, All lobes, Red, Slight 

Microscopic Observations/Comments 

Examined; 1 correlation found: 

CONGESTION, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Moderate. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, Slight. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

CELLS/INFLAMMATORY CELLS/CELL DEBRIS, 

Slight. 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC 

INFLAMMATION, Multifocal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449' 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic observations 

Page: 30 

STATUS: Humane/Otherwise Specified 

PHASE DAY OF DEATH: 1 

SEX: Female 

PHASE: Dosing phase 

ANIMAL: 1550 

GROUP,; l 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

Microscopic Observations/Comments 

CONGESTION, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

BRONCHIOLAR EPITHELIUM: DEQUAMATED, Slight. 

BRONCHIOLAR/PERIBRONCHIOLAR ACUTE/SUBACUTE 

INFLAMMATION, Slight. 

BRONCHIOLAR LUMENS: DESQUAMATED EPITHELIAL 

CELLS/INFLAMMATORY CELLS/CELL DEBRIS, 

Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......•.. No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 31 

ANIMAL;_ 2546 

GROUP: 2 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue 

Lungs ........ . 

Gross Observations/Comments 

Adhesion, Left lobe(s), Moderate/ To 

thoracic wall 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

Examined; 1 correlation found: 

VISCERAL PLEURA/CAPSULE: FIBROSIS, 

Focal, Minimal. 

HEMORRHAGE(S), Multifocal, Slight. 

Page; 32 

ANIMAL:,,2547 

GROUP: 2 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

SUBACUTE (CHRONIC ACTIVE)/CHRONIC 

INFLAMMATION, Multifocal, Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs .......•. No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 33 

ANIMAL.: 2548 

GROUP: 2 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ....•.... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 34 

ANIMAL.t 2549 

GROUP: 2 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 35 

ANIMAL: 2550 

GROUP: 2 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH: 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Female 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs . · · · · · · · · Discolored, All lobes, Red, Slight Examined; 1 correlation found: 

CONGESTION, Slight. 

ALVEOLAR/!NTRAALVEOLAR MACROPHAGES, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 36 

AN!Mb.L: 3546 

GROUP: 3 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH: 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Female 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs ........ . 

Discolored, All lobes, Red, Slight Examined; 1 correlation found: 

CONGESTION, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 37 

ANIMAL! .. 3547 

GROUP: 3 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS : ·Found Dead 

PHASE DAY OF DEATH: 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Female 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs ........ . 

Discolored, Right lobes, Red, Slight Examined; 1 correlation found: 

CONGESTION, Slight. 

HEMORRHAGE(S), Multifocal, Minimal. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 38 

ANIMAL;. 3548 

GROUP: 3 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH: 0 

.. ------. .. , 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Female 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs ....... . 

Discolored, All lobes, Red, Slight 

CONGESTION, Slight. 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 39 

ANIMAL.: 3549 

GROUP: 3 

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02-Jan-03; 18:30 

Huntingdon Life sciences, Inc. 025449 

STATUS: Found Dead 

PHASE DAY OF DEATH: 0 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

SEX: Female 

PHASE: Dosing phase 

Tissue Gross Observations/Comments Microscopic Observations/Comments 

Lungs . . . . . . . .. 

Discolored, All lobes, Red, Moderate Examined; 1 correlation found: 

CONGESTION, Moderate. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 40 

ANIMAL;, 3550 

GROUP: 3 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

Page: 41 

ANIMAL,: 4546 

GROUP: 4 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 42 

ANIMAL: 4547 

GROUP: 4 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

LYMPHOID CELL AGGREGATE(S), Multifocal, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 43 

ANIMAL: 4548 

GROUP: 4 

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02-Jan-03; 18:30 

Huntingdon Life ,sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/comments 

Lungs ......... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 44 

ANIMAL: 4549 

GROUP: 4 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue 

Lungs ........ . 

Gross Observations/Comments 

Discolored, All lobes, Red,. Foci, 0.2 - 0.5 

em 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

Examined; 1 correlation found: 

HEMORRHAGE(S), Multifocal, Slight. 

CONGESTION, Slight. 

Page: 45 

ANIMAL: 4550 

GROUP: 4 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue, 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

INTERSTITIUM: OSSEOUS METAPLASIA, Focal, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 46 

ANIMAL; 5546 

GROUP: 5 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/comments 

Lungs ......... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 47 

ANIMAL_; 5547 

GROUP: 5 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

Page: 48 

ANIMAL_:_ 5548 

GROUP: 5 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, Slight. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Observations 

STATUS: Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Page: 49 

ANIW\L: 5549 

GROUP: 5 

LYMPHOID CELL AGGREGATE(S), Focal, Minimal. 

Tissues without comment under Gross Observations were within normal limits at necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

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02-Jan-03; 18:30 

Huntingdon Life Sciences, Inc. 025449 

An Acute (4-Hour) Inhalation Toxicity Study 

in the Rat via Nose-Only Exposure 

Individual Animal Gross and Microscopic Ooservations 

STATUS: ·Final phase sacrifice 

PHASE DAY OF DEATH: 14 

Tissue Gross Observations/Comments 

Lungs ......... No gross observations on tissue. 

SEX: Female 

PHASE: Dosing phase 

Microscopic Observations/Comments 

ARTERIES: MEDIAL HYPERTROPHY, Minimal. 

ALVEOLAR/INTRAALVEOLAR MACROPHAGES, 

Minimal. 

Tissues without comment under Gross Observations were within normal limits at.necropsy. 

The following tissues were unremarkable microscopically: 

No tissues examined. 

Page: 50 

ANIMA4; 5550 

GROUP: 5 

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Huntingdon Life Sciences 02-5449 

Equipment List 

Exposure Chamber 

Page 140 

Final Report 

Appendix A 

Flow-past nose-only chamber with polycarbonate nose-only tubes (CR Equipment, SA). 

Compound Generator 

Glass Bubbler (Crown Glass Co., Inc.). 

Flowmeter 

Dwyer®, sizes 0-0.24, 0-1, 0-4, 0-10 and 0-20 Lpm (Dwyer® Instruments), calibrated prestudy with 

a Sierra Instruments Top Trak Mass Flowmeter, Modei822-13-0VI-PVI-VI. 

Pressure Gauges 

Norgreen backpressure gauge, P/N 9892K23. 

Dwyer® Magnehelic® gauge, Model N/A. 

U.S. Gauge backpressure gauge, P/N 12672-1. 

Regulators 

Norgreen, P/N 9892KO and 9892K23. 

Union Carbide, P/N SG380030. 

Metering Valve 

Nupro® Co., Model SS-4L Series. 

Tubing 

Clear plastic size 1/4", 1/2", 1" (Norton). 

Teflon® size 1/8", 1/4". 

Stainless steel size 1/2". 

Stainless steel "T" tube, W' 

Air Analyzer 

MIRAN® 1A Ambient Air Analyzer (Wilks) with a Cole Parmer Strip Chart Recorder, Model 201, 

and a Micronta® LCD Benchtop Digital Multimeter No. 22-195. 

Balston® MicrofibreTM Disposable Filter Units were attached in-line. 

Particle Sizer 

TSI Aerodynamic Particle Sizer, Model 330000, and a DELL computer, Model 486P/25, equipped 

with an Epson LQ-500 Dot matrix printer, Model P78PA. 

Thermometer and Humidity Gauge 

Springfield and Sunbeam Temperature and Humidity Gauges, calibrated prestudy with a VWR 

Digital Hygrometer and Thermometer. 

Balances 

AND EP12KA. 

AND FX 6000. 

Mettler PE11. 
Huntingdon Life Sciences 02-5449 

MIRAN® Calibration 

Methodology for 2-Bromo-3,3,3-Trifluoropropene 

Page 141 

Final Report 

Appendix 8 

Settings: The instrument settings for the unit are summarized below: 

Wavelength, microns 3.42 

Pathlength, dial setting 14.00 

Slit width, mm 1 

Range,absorbance 1 

Response,seconds 1 

Gain High 

Chart speed, em/min 1 

Chart volts 1 

Calibrations: The MIRAN® was turned on and allowed to warm up for 

approximately 1 0 minutes. The cell was flushed with room air for approximately 

one minute. The loop was closed, the unit was zeroed and the calibration series 

was performed as shown below. The resultant data were plotted to obtain a 

calibration curve. Each observer used a separate syringe for calibration. 

Injection Calculated Absorbance 

Volume Concentration 1 O~erator 1 O~erator 2 Average 

(til) (ppm) (volts) (volts) (volts) 

Group 1 

510 20,000 0.433 0.474 0.454 

640 25,120 0.553 0.571 0.562 

760 29,830 0.634 0.661 0.648 

Group 4 

100 3,925 0.1292 0.1221 0.1257 

130 5,102 0.1386 0.1390 0.1388 

160 6,280 0.1658 0.1686 0.1672 

1

Calculated Cone. (ppm)= 

Volume (~L) x Density (mg/~L) x (1000 ~g/mg) x (22.4 ~U~mole) x (295K/273K} 

MW (pg/Jlmole) x 5.64L 

where: 5.64L =volume of MIRAN® closed-loop 

Density (mg/JIL) = 1.60 

MW (tig/Jimole) = 174.95 
Huntingdon Life Sciences 02-5449 

MIRAN® Calibration 

Page 142 

Final Report 

Appendix B 

Calibration Checks: A one-point calibration check of the MIRAN® was performed 

prior to each exposure. The parameters are shown below: 

Injection 

Volume 

(pl) 

Group 1 

640 

Group 4 

130 

Calculated 

Concentration 

(ppm) 

25,120 

5,102 

Expected Acceptable 

Absorbance Absorbance 

Reading Range 

(volts) (volts) 

0.562 0.478- 0.646 

0.1388 0.1180-0.1596 

The absorbance was recorded after each injection. The absorbance was 

considered satisfactory if it was within 15% of the original calibration series. If 

the absorbance value fell outside the 15% range, the injection was rechecked as 

follows. The volume for the value that was out of range was reinjected twice. 

The closer pair of the three injections were averaged and the results were 

compared to the original curve. If the average of the pair was within the 15% 

range, the original was accepted. If the value of the average was outside the 

15% range, the Study Director decided if a new graph was to be prepared. 
Huntingdon Life Sciences 02-5449 

MIRAN® Calibration 

Calibration CuNe for 2-Bromo-3,3,3-Trifluoropropene 

Group 1 

Page 143 

Final Report 

Appendix B 

31000~--------------------------------------~ 

,.... 

30000 

29000 

28000 

a. 27000 

c. 

';;" 26000 

0 'i 25000 ... 

-

§ 24000 

(.) 8 23000 

22000 

21000 

3 2 

y = 651 07x - 64586x + 60360x 

R2 =0.9666 

• 19000+-------+-------4-------~-------+-------4 

0.430 0.480 0.530 0.580 0.630 0.680 

Absorbance (Volts) 
Huntingdon Life Sciences 02-5449 Page 144 

Final Report 

MIRAN® Calibration Appendix B I 

Calibration Curve for 2-Bromo-3,3,3-Trifluoropropene 

Group 4 

6500~------------------------------------~ 

6000 

,......., 

a. 5500 

c. 

"'-" 

c 

0 'i 5000 .... 

-

c 

Q) 

(..) 

§ 4500 

u 

4000 

• y =-6142817.5944i + 1933968.2043l-114025.3737x 

R2 =0.9636 

3500+-----------~-------------r----------~ 

0.1200 0.1400 0.1600 0.1800 

Absorbance (Volts) 
Huntingdon Life Sciences 02-5449 

MIRAN® Calibration 

Methodology for E-1-Bromo-3,3,3-Trifluoropropene 

Page 145 

Final Report 

Appendix B 

Settings: The instrument settings for the unit are summarized below: 

Wavelength, microns 3.44 

Pathlength, dial setting 8.17 

Slit width, mm 1 

Range,absorbance 1 

Response,seconds 1 

Gain High 

Chart speed, em/min 1 

Chart volts 1 

Calibrations: The MIRAN® was turned on and allowed to warm up for 

approximately 1 0 minutes. The cell was flushed with room air for approximately 

one minute. The loop was closed, the unit was zeroed and the calibration series 

was performed as shown below. The resultant data were plotted to obtain a 

calibration curve. Each observer used a separate syringe for calibration. 

Injection 

Volume 

(/JL) 

Group 2 

490 

620 

740 

Group 5 

1100 

1250 

1400 

Calculated 

Concentration 1 

(ppm) 

19,830 

25,100 

29,950 

44,524 

50,595 

56,666 

1

Calculated Cone. (ppm)= 

OQerator 1 

(volts) 

0.586 

0.691 

0.765 

0.662 

0.726 

0.770 

Absorbance 

OQerator 2 

(volts) 

0.605 

0.710 

0.785 

0.667 

0.719 

0.766 

Average 

(volts) 

0.596 

0.701 

0.775 

0.665 

0.723 

0.768 

Volume (yl) x Density (mg/yl) x (1000 yg/mg) x (22.4yUymole) x (295K/273K) 

MW (.ug/,umole) x 5.64L 

where: 5.64L =volume of MIRAN® closed-loop 

Density (mg/pL) = 1.65 

MW (/Jg/pmole) = 174.95 
( ·~ .. \:· .·· 

Huntingdon Life Sciences 02-5449 

MIRAN® Calibration 

Page 146 

Final Report 

Appendix B 

Calibration Checks: A one-point calibration check of the MIRAN® was performed 

prior to each exposure. The parameters are shown below: 

Expected Acceptable 

Injection Calculated Absorbance Absorbance 

Volume Concentration Reading Range 

{til) (ppm) (volts) (volts) 

Group 2 

620 25,100 0.701 0.596- 0.806 

Group 5 

1250 50,595 0.723 0.615- 0.831 

The absorbance was recorded after each injection. The absorbance was 

considered satisfactory if it was within 15% of the original calibration series. If 

the absorbance value fell outside the 15% range, the injection was rechecked as 

follows. The volume for the value that was out of range was reinjected twice. 

The closer pair of the three injections were averaged and the results were 

compared to the original curve. If the average of the pair was within the 15% 

range, the original was accepted. If the value of the average was within the 15% 

range, the original was accepted. If the value of the average was outside the 

15% range, the Study Director decided if a new graph was to be prepared. 
Huntingdon Life Sciences 02-5449 

MIRAN® Calibration 

Page 147 

Final Report 

Appendix B 

Calibration Curve for E-1-Bromo-3,3,3-Trifluoropropene 

Group 2 

31000~---------------------------------------, 

,...., 

30000 

29000 

28000 

a. 27000 

c. 

"';; 26000 

0 'i 25000 .... 

-

~ 24000 

(.) 

c 

0 23000 

u 

22000 

21000 

3 2 

y = 471 08x - 36268x + 38259x 

R2 =0.9813 

20000 

19000+-------+-------~-------r-------+------~ 

0.550 0.600 0.650 0.700 0.750 0.800 

Absorbance (Volts) 
( 

Huntingdon Life Sciences 02-5449 

Ml RAN® ·calibration 

Page 148 

Final Report 

Appendix B 

Calibration Curve for E-1-Bromo-3,3,3-Trifluoropropene 

Group 5 

60000,--------------------------------------, 

59000 

58000 

57000 

56000 

E' 55ooo 

0. 54000 

a 53ooo 

5 52000 

'i 51000 

-E 50000 

~ 49000 

5 48000 

(.) 47000 

46000 

45000 

44000 

3 2 

y = 287208x - 346602x + 170531 x 

R2 =0.9958 

43000 

42000+-------+-------r-------r-------~----~ 

0.550 0.600 0.650 0.700 0.750 0.800 

Absorba nee (Volts) 
Huntingdon Life Sciences 02-5449 

MIRAN® Calibration 

Page 149 

Final Report 

Appendix B 

Methodology for 1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene 

Settings: The instrument settings for the unit are summarized below: 

Wavelength, microns 

Pathlength, dial setting 

Slit width, mm 

Range, absorbance 

Response, seconds 

Gain 

Chart speed, em/min 

Chart volts 

3.42 

12.86 

1 

1 

1 

High 

1 

1 

Calibrations: The MIRAN® was turned on and allowed to warm up for 

approximately 10 minutes. The cell was flushed with room air for approximately 

one minute. The loop was closed, the unit was zeroed and the calibration series 

was performed as shown below. The resultant data were plotted to obtain a 

calibration cuNe. Each obseNer used a separate syringe for calibration. 

Injection Calculated Absorbance 

Volume Concentration 1 O~erator 1 O~erator 2 Average 

(pl) (ppm) (volts) (volts) (volts) 

Group 3 

620 19,820 0.581 0.583 0.582 

780 24,940 0.706 0.685 0.696 

940 30,060 0.796 0.782 0.789 

1

Calculated Cone. (ppm)= 

Volume (yl) x Density (mg/yl) x (1000 yg/mg) x (22.4 yUymole) x (295K/273K) 

MW (pg/,umole) x 5.64L 

where: 5.64L =volume of MIRAN® closed-loop 

Density (mg/,ul) = 1.81 

MW (pg/,umole) = 242.95 
Huntingdon Life Sciences 02-5449 

MIRAN® Calibration 

Page 150 

Final Report 

Appendix 8 

Calibration Checks: A one-point calibration check of the MIRAN® was performed 

prior to the exposure. The parameters are shown below: 

Expected Acceptable 

Injection Calculated Absorbance Absorbance 

Volume Concentration Reading Range 

{til) (ppm) (volts) (volts) 

Group 3 

780 24,940 0.696 0.592 - 0.800 

The absorbance was recorded after each injection. The absorbance was 

considered satisfactory if it was within 15% of the original calibration series. If 

the absorbance value fell outside the 15% range, the injection was rechecked as 

follows. The volume for the value that was out of range was reinjected twice. 

The closer pair of the three injections were averaged and the results were 

compared to the original curve. If the average of the pair was within the 15% 

range, the original was accepted. If the value of the average was outside the 

15% range, the Study Director decided if a new graph was to be prepared. 
Huntingdon Life Sciences 02-5449 

MIRAN® Calibration 

Page 151 

Final Report 

Appendix B 

Calibration Curve for 1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene 

Group 3 

31000~------------------------------------~ 

-

30000 

29000 

28000 

a. 27000 

c. 

~ 26000 

0 'i 25000 ... -

~ 24000 

(.) 8 23000 

22000 

21000 

3 2 

y = 26543x - 17 422x + 35223x 

R2 =0.9916 

20000 

19000+----+----+----r----+----+----+----+--~ 

0.430 0.480 0.530 0.580 0.630 0.680 0.730 0.780 0.830 

Absorbance (Volts) 
Appendix C 

PROTOCOL 

2-Bromo-3,3,3-Trifluoropropene, 

1-Bromo-3,3,3-Trifluoropropene and 

Page 152 

1-Bromo-2-Trifluoromethyl -3,3,3-Trifluoropropene 

AN ACUTE (4-HOUR) INHALATION TOXICITY STUDY 

IN THE RAT VIA NOSE-ONLY EXPOSURE 

HLS Study No.: 

Protocol No.: 

Date: 

02-5449 

Final 

10 July 2002 
·PROTOCOL SIGNATURES I PREFACE 

(Confidential Information- to be distributed on a need-to-know basis) 

Study Title: 2-Bromo-3,3,3-Trifluoropropene, 

1-Bromo-3,3,3-Trifluoropropene and 

Page 153 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene: 

An Acute (4-Hour) Inhalation Toxicity Study in the Rat 

via Nose-Only Exposure 

HLS Study No.: 02-5449 

This is the Final Protocol. It has been reviewed and approved by: 

Gary . Ho an, .A., DABT 

Study Direct r 

Huntingdon Life Sciences 

Mettlers Road 

East Millstone, NJ 08875-2360 

Phone No.: 1-732-873-2550 x2920 

Fax No.: 1-732-873-3992 

Email: hoffmang@princeton.huntingdon.com 

B~- Sponsor Representative 

. American Pacific Corporation 

3770 Howard Hughes Parkway 

Suite300 

Las Vegas, NV 89109 

PhoneNo.: 1-702-699-4131 

Fax No.: 1-702-735-4876 

Email: bcolton@apfc.com 

!3 ~ 2002.. 

Date 
Page 154 

TABLE OF CONTENTS 

1.. INTRODUCTION .............................. .-...................................................................... 1 

2. STUDY PERSONNEL ........................................................................................... l 

3. PROPOSED STUDY DATES ............................................................................... l 

4. . EXPERIMENTAL DESIGN ................................................................................. 2 

4.1. JUSTIFICATION FOR ROUTE, DURATION AND FREQUENCY .................... 2 

4.2. JUSTIFICATION FOR TEST ANIMAL SELECTION ......................................... .3 

4.3. JUSTIFICATION FOR NUMBER OF ANIMALS ................................................ .3 

4.4. JUSTIFICATION FOR EXPOSURE LEVEL SELECTION ................................. .3 

5. TEST SUBSTANCES ............................................................................................ 3 

5.1. TEST SUBSTANCES: ............................................................................................ 3 

5.2. IDENTIFICATION OF TEST SUBSTANCES ...................................................... .3 

5.3. ARCHIVAL SAMPLES .......................................................................................... 3 

5.4. ADMINISTRATION OF TEST SUBSTANCES ..... : ............................................. .4 

5.5. EXPOSURE CONCENTRATION DETERMINATION ....................................... .4 

5.6. PARTICLE SIZE DISTRIBUTION ANALYSIS: .................................................. .4 

5.7. CHAMBER ENVIRONMENT: .............................................................................. 5 

5.8. SUMMARY OF CHAMBER ACTIVITY: ............................................................. 5 

5.9. FREQUENCY AND DURATION OF ADMINISTRATION ................................ .5 

5.10. UNUSED TEST SUBSTANCES .......................................................................... 5 

6. TEST ANIMALS ..................................................................................................... 5 

6.1. SPECIES .................................................................................................................. 5 

6.2. SUPPLIER ............................................................................................................... 6 

6.3. ANIMAL REQUIREMENTS/SPECIFICATIONS ................................................. 6 

6.4. ACCLIMATION PERIOD ....................................... ; .............................................. 6 

6.5. ANIMAL CARE AND IIUSBANDRY ................................................................... 6 

7. IN-LIFE EVALUATIONS ...................................................................................... 9 

7.1. CLINICAL OBSERVATIONS ................................................................................ 9 

7.2. BODY WEIGHTS ................................................................................................. 10 

8. POSTMORTEM EV ALUATIONS .................................................................... lO 

8.1. MACROSCOPIC PATHOLOGY .......................................................................... lO 

8.2. METHOD OF EUTHANASIA .............................................................................. 10 

8.3. TISSUE PRESERVATION ................................................................................... 10 

8.4. ORGAN WEIGHTS .............................................................................................. 10 

8.5. MICROSCOPIC PATHOLOGY EVALUATIONS .............................................. 11 

9. ARCHIVING OF RECORDS AND SPECIMENS ........................................... ll 
Page 155 

10. STATISTICAL ANALYSIS ............................................................................... 12 

. 11. REPORTING ....................................................................................................... 12 

11.1. STATUS REPORTS ........................................................................................... 12 

11.2. FINAL REPORT ................................................................................................. 12 

12. REGULATORY REFERENCES ....................................................................... 13 

12.1. TESTWG GUIDELWES .................................................................................... 13 

12.2. GOOD LABORATORY PRACTICES ............................................................... 13 

12.3. ANIMAL WELFARE ACT COMPLIANCE ..................................................... 13 

12.4. INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE (IACUC) ...... .14 

13. QUALITY ASSURANCE MONITORING ....................................................... 14 

14. ALTERATION IN STUDY DESIGN ................................................................ 14 
Huntingdon Life Sciences 

Study No. 02-5449 

1. INTRODUCTION 

HLS Study No. 

2. 

Protocol 

Study Title 

Testing Facility 

Purpose 

STUDY PERSONNEL 

Study Director 

Alternate Contact 

Page 156 

Page 1 

Final Protocol 

02-5449 

Final 

2-Bromo-3,3,3-Trifluoropropene, 

1-Bromo-3,3,3-Trifluoropropene and 

1-Bromo-2-Trifluoromethyl-3,3,3-

Trifluoropropene: An Acute (4-Hour) 

Inhalation Toxicity Study in the Rat via 

Nose-Only Exposure 

Huntingdon Life Sciences 

Mettlers Road 

P.O. Box 2360 

East Millstone, NJ 08875-2360 

This study is designed to assess the 

potential and comparative acute inhalation 

toxicity of 3 test substances when 

administered for 4 hours via nose-only 

exposure. 

Gary M. Hoffman, B.A., DABT 

Carol S. Auletta, M.B.A., DABT x2960 

Additional personnel will be documented in the study file and presented in the 

final report. 

3. PROPOSED STUDY DATES 

Study Initiation 

Receipt of Test Animals 

First Day of Treatment 

Last Day of Treatment 

Necropsy- Termination 

Submission of Audited Draft Report 

Experimental Termination Date 

Study Completion 

Date Study Director signs protocol 

September 2002 

September 2002 (experimental start date) 

October 2002 

October 2002 

December 2002 

January 2003 (date of last data collection) 

Date final report is signed by Study 

Director 
Huntingdon Life Sciences 

Study No. 02-5449 

4. EXPERIMENTAL DESIGN 

Page 157 

Page2 

Final Protocol 

Initially, one group per test substance (consisting of five per sex per group of 

Sprague-Dawley rats) will be exposed to an atmosphere containing the test 

substance for four hours at a measured concentration of approximately 2.5% which 

is equivalent to 25,000 ppm (parts per million of test substance in air) or at a 

maximum attainable concentration, whichever is lower. These animals will then be 

held for a 14-day post-exposure observation period. If signs of apparently reversible 

toxicity still persist, they will be held for one additional 7 -day observation period. 

At the end of the observation period or upon spontaneous death, a macroscopic 

necropsy examination, measurement of selected organ weights and a microscopic 

examination of the lungs will be performed on all animals. 

If any compound-related mortality occurs in the initial group( s ), additional testing to 

estimate the median lethal concentration may be performed (Sponsor consulted). At 

least one additional group consisting of five of the more sensitive sex as determined 

by the initial test group of Sprague-Dawley rats each will be exposed to atmosphere 

containing the test substance for four hours. If the more sensitive sex cannot be 

determined, then five per sex will be exposed for the additional group(s). The target 

concentration(s) will be selected to try to produce at least two groups with partial 

mortality. Exposure levels for the additional group(s) will be documented in the 

raw data and final report. The animals will then be held for a 14-day post-exposure 

observation period. If signs of apparently reversible toxicity still persist, they will 

be held for one additional 7-day observation period. At the end of the observation 

period or upon spontaneous death, a macroscopic necropsy examination, 

measurement of selected organ weights and a microscopic examination of the lungs 

will be performed on all animals. Estimates of the median lethal concentration, with 

95% confidence limits, will be calculated where appropriate. A control exposure 

group is considered unnecessary unless a vehicle (other than water) is used for test 

substance administration. Where adequate information is available to demonstrate 

that animals of the sex tested are markedly more sensitive, testing in animals of the 

other sex is not required. 

4.1. JUSTIFICATiON FOR ROUTE, DURATION AND FREQUENCY 

The inhalation route is one of the potential routes of human exposure to 

this test substance and is the route as per the regulatory requirements of 

the OECD and EPA as indicated in Section 12.1. The duration of the 

study and frequency of exposures are as per the regulatory requirements of 

the OECD and EPA as indicated in Section 12.1. 
1/ 

Page 158 

Huntingdon Life Sciences Page 3 

Study No. 02-5449 Final Protocol 

4.2. JUSTIFICATION FOR TEST ANIMAL SELECTION 

The rat is a rodent animal model commonly utilized in toxicity testing as 

per the regulatory requirements of the OECD and EPA as indicated in 

Section 12.1. ill addition, a historical database is available for 

comparative evaluation. 

4.3. JUSTIFICATION FOR NUMBER OF ANIMALS 

The number of animals in the protocol is considered to be the minimum 

necessary for scientific and statistical reasons in order to evaluate the 

data with sufficient confidence levels as per the regulatory requirements 

of the OECD and EPA as indicated in Section 12.1. 

4.4. JUSTIFICATION FOR EXPOSURE LEVEL SELECTION 

The exposure levels are as specified by the Sponsor based on prior testing 

of these or similar chemicals. 

5. TEST SUBSTANCES 

5.1. TEST SUBSTANCES: 

. 2-Bromo-3,3 ,3-Trifluoropropene, 

1-Bromo-3,3,3-Trifluoropropene and 

1-Bromo-2-Trifluoromethyl -3,3,3-Trifluoropropene 

Test Substances Category: industrial gases 

Description, lot number, storage, expiration date (if available) and 

handling procedures, as well as other pertinent information will be 

documented in the study data. 

5.2. IDENTIFICATION OF TEST SUBSTANCES 

Unless otherwise noted, the identity, strength, purity, composition, 

stability and method of synthesis, fabrication and/or derivation of each 

batch ofthe test substances will be documented by the Sponsor before its 

use in the study. This documentation will be maintained by the Sponsor. 

5.3. ARCHlY AL SAMPLES 

An archival sample from each lot of test substance will NOT be taken 

and stored in the Archives of the Testing Facility. 
Huntingdon Life Sciences 

Study No. 02-5449 

5.4. ADMINISTRATION OF TEST SUBSTANCES 

Page 159 

Page4 

Final Protocol 

The test substances will be administered by nose-only inhalation 

exposure. The test substances will be administered as a vapor (depending 

on the physical properties of the test substance) in the breathing air of the 

animals. The test atmospheres will be generated by an appropriate 

procedure determined during pre-study trials. The trials will be 

performed to evaluate the optimal set of conditions and equipment to 

generate stable atmospheres at the target exposure level(s). The method 

will be described in the raw data of the study and in the report. · 

The flow-past nose-only exposure chambers will be operated at a 

minimum flow rate of 8 liters per minute. The final airflow will be set to 

provide at least one air change in 5.0 minutes (12 air changes/hour) and a 

T 99 equilibrium time of at most 23 minutes. The T 99 equilibrium time 

will be added to the 4 hours; i.e., the atmosphere will be generated for 4 

hours and T 99 minutes. This chamber size and airflow rate is considered 

adequate to maintain the oxygen level at least 19%. At the end of the 

exposure, all animals will remain in the chamber for a minimum of 30 

minutes. During this time the chamber will be operated at approximately 

the same flow rate using clean air only. 

5.5. EXPOSURE CONCENTRATION DETERMINATION 

A nominal exposure concentration will be calculated if possible. The 

flow of air through the chamber will be monitored using appropriate 

calibrated equipment. The test substance consumed during the exposure 

will be divided by the total volume of air passing through the chamber 

(volumetric flow rate times total exposure time) to give the nominal 

concentration. 

During each exposure, measurements of airborne concentrations of test 

substance will be performed each hour using an infrared (IR) 

spectrophotometric sampling procedure. 

If more than the normal amount of trial time is required because of test 

substance generation or monitoring problems ( 40 technician hours), the 

Sponsor will be consulted prior to additional trials (additional cost). 

5.6. PARTICLE SIZE DISTRIBUTION ANALYSIS: 

During each exposure, particle size determinations will be performed each 

hour using a cascade impactor or other appropriate device, to characterize 

the aerodynamic particle size distribution of any aerosol present. 
Huntingdon Life Sciences 

Study No. 02-5449 

5.7. CHAMBER ENVIRONMENT: 

Paqe 160 

¥age 5 

Final Protocol 

Temperature, humidity and airflow rate will be recorded every 30 minutes 

during exposure. Chamber temperature and relative humidity will be 

maintained, to the extent possible, between 20 to 24 oc and 30 to 70%, 

respectively. 

5.8. SUMMARY OF CHAMBER ACTIVITY: 

The minimum frequency of chamber activity is summarized below: 

Activity 

Measured Test Substance Concentration 

Particle Size 

Temperature 

Relative Humidity 

Airflow Rate 

Nominal Test Substance Concentration 

Loading/Unloading Verification 

Frequency/Exposure 

4X 

lX 

9X 

9X 

9X 

IX 

lX 

5.9. FREQUENCY AND DURATION OF ADMINISTRATION 

The test substances will be administered for 4 hours for 1 day. 

5.10. UNUSED TEST SUBSTANCES 

The unused portion of the test substances as well as any erilpty test 

substance containers will be returned to the Sponsor following 

completion of the in-life phase of the study. In the event the Sponsor 

wishes the Testing Facility to arrange for disposal, a cost for this service 

will be provided. 

6. TEST ANIMALS 

6.1. SPECIES 

Albino Rats (Outbred) Va:f/Plus® 

Sprague-Dawley- derived (CD®) 

Crl: CD (SD) IGS BR 
Page 161 

Huntingdon Life Sciences Page 6 

Study No. 02-5449 Final Protocol 

6.2. SUPPLIER 

Charles River Laboratories 

Documentation of the specific breeding facility will be maintained in the 

study file. 

6.3. ANIMAL REQUIREMENTS/SPECIFICATIONS 

6.3.1. NUMBER OF ANIMALS ON STUDY 

5 per sex per test substance for the initial groups and 5 per sex per 

test substance or 5 of one sex per test substance, as determined, 

for the subsequent groups with a minimum of 1 group and a 

maximum of 6 groups will be used. Should additional groups be 

required, they will be added following consultation with the 

Sponsor. 

6.3.2. AGE 

Young adults (approximately 7-11 weeks old at receipt and 

between 8-12 weeks old at exposure initiation). 

6.3.3. WEIGHT 

Males: Approximately 200-360 grams at initiation 

Females: Approximately 180-260 grams at initiation 

Individual weight should not exceed ± 20% of the mean weight 

for each sex. Animals outside of these ranges may be used at 

the discretion of the Study Director. 

Note: Females will be nulliparous and nonpregnant. 

6.4. ACCLIMATION PERIOD 

Approximately 1 week; all animals will be checked for viability twice 

daily. Prior to assignment to study, all animals will be examined to 

ascertain suitability for study. 

6.5. ANIMAL CARE AND HUSBANDRY 

6.5.1. FACILITIES MANAGEMENT/ANIMAL HUSBANDRY 

Currently acceptable practices of good animal husbandry will be 

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

Animals; National Academy Press, 1996. Huntingdon Life 

Sciences Inc. is fully accredited by the Association for 
Huntingdon Life Sciences 

Study No. 02-5449 

Page 162 

Page7 

Final Protocol 

Assessment and Accreditation of Laboratory Animal Care 

International (AAALAC). 

6.5.2. VETERINARY CARE 

Animals will be monitored by the technical staff for any 

conditions requiring possible veterinary care. If any such 

conditions are identified, a staff veterinarian will be notified for 

an examination and evaluation. Animals will be treated as 

outlined in the Animal Welfare Act Compliance section of this 

protocol. 

6.5.3. ENVIRONMENTAL CONDITIONS 

Light/Dark Cycle 

Twelve hour light/dark cycle provided via automatic timer. 

Temperature 

Temperature will be monitored in accordance with Testing 

Facility SOPs to ensure that the desired range of 18 to 26°C is 

maintained to the maximum extent possible . 

. Humidity 

Humidity will be monitored in accordance with Testing Facility 

SOPs to ensure that the desired range of 30 to 70% IS 

maintained to the maximum extent possible. 

6.5.4. HOUSING 

Group housed (2-6/cage) during acclimation, then individually in 

suspended stainless steel wire mesh cages during study. 

6.5.5. FEED 

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

International, St. Louis, MO) ad libitum. 

6.5.6. WATER 

Elizabethtown Water Company, Westfield, NJ; ad libitum, via 

automated watering system. 
Huntingdon Life Sciences 

Study No. 02-5449 

6.5.7. FEED ANALYSIS 

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Final Protocol 

Analytical certification of batches of feed provided by the 

manufacturer will be maintained on file by the manufacturer. 

There are no known contaminants in the feed which are 

expected to interfere with the objectives of this study. 

6.5.8. WATER ANALYSIS 

Water analyses are conducted by Elizabethtown Water 

Company to assure that water meets standards specified under 

the EPA Federal Safe Drinking Water Act Regulations (40 CFR 

Part 141). Water analysis, provided by the supplier, will be 

maintained on file at the Testing Facility. In addition, chemical 

and microbiological analyses are conducted biannually on water 

samples collected from representative rooms in this facility. 

Results are maintained on file. There are no known 

·contaminants which are expected to interfere with the objectives 

ofthis study. 

6.5.9. ANIMAL ASSIGNMENT 

More animals than required for the study will be purchased and 

acclimated. Animals will· be arbitrarily placed in cages upon 

receipt and will be placed on study as available at time of study 

initiation. Animals considered unsuitable for study on the basis 

of pretest physical examinations and outlying body weights will 

be excluded. Disposition of all animals not utilized in the study 

will be maintained on file at the Testing Facility. 

6.5.10. IDENTIFICATION 

Each animal will be assigned a temporary identification number 

upon receipt. After selection for study, each animal will be eartagged with a number assigned by the Testing Facility. This 

number plus the study number will comprise the unique animal 

number for each animal. If the tag is lost, it will be replaced. 

Each cage will be provided with a cage card which will be color 

coded for exposure level identification and will contain the 

study number and animal number. 

6.5.11. ANIMAL HUSBANDRY/EXPOSURE 

Housing: Individually in tubes. 
Huntingdon Life Sciences 

Study No. 02-5449 

Food: 

Water: 

7. IN-LIFE EVALUATIONS 

7.1. CLINICAL OBSERVATIONS 

None. 

None. 

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Final Protocol 

NOTE: Animals showing severe and enduring signs of distress and pain may need 

to be humanely euthanized as per OECD and EPA test guidelines. 

7.1.1. VIABILITY CHECKS (CAGE-SIDE) 

Observations for mortality and general condition will be made 

at least twice daily (once in the morning and once in the 

afternoon). Animals in poor health or in a possible moribund 

condition will be identified for further monitoring and possible 

euthanasia. 

7.1.2. PHYSICAL EXAMINATIONS 

Day 0 (Day ofExposure): Animals will be observed individually 

prior to exposure, as a group at fifteen minute intervals during the 

first hour of exposure, and hourly for the remainder of the 

exposure. They will be observed individually upon removal from 

the chainber and hourly for two hours post -exposure. 

Days 1-14: Detailed general observations will be recorded once 

daily. Ifthe animals are held for an additional one-week recovery 

period, they will be observed from Day 15 through Day 21. 

Examinations will include but not be limited to, evaluation of 

skin and fur, eyes and mucous membranes, respiratory and 

circulatory effects, autonomic effects such as salivation, central 

nervous system effects, including tremors and convulsions, 

changes in the level of motor activity, gait and posture, reactivity 

to handling or sensory stimuli, grip strength, and stereotypies or 

bizarre behavior (e.g., self- mutilation, walking backwards). 

Detailed observations will be recorded at each interval. 
Huntingdon Life Sciences 

Study No. 02-5449 

7.2. BODYWEIGHTS 

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Final Protocol 

Day 0 (prior to exposure), Days 7 and 14 (termination). Ifthe animals are 

held for an additional one week recovery period, they will also be weighed 

on Day 21 (Termination). 

8. POSTMORTEM EVALUATIONS 

8.1. MACROSCOPIC PATHOLOGY 

A complete macroscopic examination will be performed on all animals, 

including animals euthanized in a moribund condition or found dead; all 

abnormal observations will be recorded. The necropsy will include 

examination of the external surface and all orifices; the organs and tissues 

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

remainder of the carcass. Tissues and lesions will be saved as indicated in 

Section 8.3. 

8.2. METHOD OF EUTHANASIA 

Exsanguination following carbon dioxide inhalation. 

8.3. TISSUE PRESERVATION 

All weighed organs as listed in Section 8.4 will be preserved in 10% 

neutral buffered formalin. The lungs will be infused with 10% neutral 

buffered formalin to ensure fixation and preserved for all animals. 

Additionally, if macroscopically observed pathological lesions are 

observed in animals which survive more than 24 hours, the lesion will be 

taken at necropsy and preserved in 10% neutral buffered formalin. A 

pathologist will review the data to determine which lesions appear to be 

due to the test substance. Lesions which appear to be due to the test 

substance will be archived, or examined microscopically (additional cost), 

as determined by the Sponsor 

8.4. ORGAN WEIGHTS 

Organs indicated below will be taken from all survivors at the scheduled 

necropsies, weighed, recorded and organ/body and organ/brain weight 

ratios calculated. Organs will not be weighed for animals found dead or 

euthanized in a moribund condition during the course of the study. Prior 

to weighing, all organs will be carefully dissected and properly trimmed 

to remove fat and other contiguous tissues in a uniform manner. Organs 

will be weighed as soon as possible after dissection to avoid drying. 

Paired organs will be weighed together. 
( 

Huntingdon Life Sciences 

Study No. 02-5449 

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Final Protocol 

9. 

adrenals 

brain 

kidneys 

lungs 

liver 

ovanes 

testes 

8.5. MICROSCOPIC PATHOLOGY EVALUATIONS 

For all animals in all groups, the lungs will be examined microscopically. 

Tissues with macroscopic lesions and target organs may be examined in 

all animals (additional cost, Sponsor consulted). Target organs will be 

designated by the Study Director, pathologist and/or Sponsor based on 

experimental findings. Authorization will be obtained from the Sponsor 

prior to performing additional examinations. Any abnormalities not 

noted during macroscopic postmortem examinations which are seen 

during histological processing will be recorded. Additional examinations 

will be made only after consultation with the Sponsor and receipt of 

authorization from the Sponsor. 

Stains 

Standard stains will be used (hematoxylin and eosin). Special stains may 

be employed on selected tissues in making a diagnosis at the discretion 

of the Study Pathologist. Special stains may be employed at the request 

of the Sponsor (additional cost). 

ARCHIVING OF RECORDS AND SPECIMENS 

All data documenting experimental details and study procedures and observations 

will be recorded and maintained as raw data. At the completion of the study, all 

reports, raw data, preserved archival specimens and retained samples will be 

maintained in the Testing Facility's Archives for a period of 1 year after 

submission of the signed final report. 

The Sponsor will be contacted in order to determine the fmal disposition of these 

materials. The Sponsor is responsible for all cost associated with the storage ofthese 

materials beyond one year from the issuance of the final report and for any costs 

associated with the shipment of these materials to the Sponsor or to any other 

facility designated by the Sponsor. 
Huntingdon Life Sciences 

Study No. 02-5449 

10. STATISTICAL ANALYSIS 

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Final Protocol 

If appropriate, a calculation of an estimate of the median lethal concentration, with 

95% confidence limits, using the method of Maximum Likelihood Estimation1 

or 

Litchfield and Wilcoxon2 will be performed for the combined sexes, the males and 

the females. 

1 Finney, D.J. Probit Analysis, 3rd Edition, Cambridge University Press, London, 1971. 

2 Litchfield, J.T., Jr. and F. Wilcoxon, 1949, J. Pharma. Exp. Therap., 96:99-113. 

11. REPORTING 

11.1. STATUS REPORTS 

Periodic verbal and written updates on study progress will be provided 

by the Study Director. 

11.2. FINAL REPORT 

One copy of an audited draft report will be submitted following 

termination of the study. After receipt and review of the Sponsor's 

comments, appropriate changes will be made and 2 paper copies of a 

signed, audited final report will be issued. (Additional copies of reports 

will be provided at additional cost). The report will include, but not be 

limited to: 

Compliance Statement 

Quality Assurance Statement 

Summary 

Introduction 

Experimental Design 

Materials and Methods 

Discussion of Study Results 

Conclusion 

Tables of Exposure Data 

Mortality Data with LC50 (if applicable) 

Tables of individual and summary daily physical examinations 

Tables of individual body weights and body weight changes 

Table of individual macroscopic postmortem fmdings (lesions which 

appear to be due to test substance will be noted) 

Table of individual organ weights and organ weight ratios 

Table of individual microscopic postmortem findings (as appropriate) 

List of senior personnel participating in the study 

Copy of study protocol and associated amendments 
Huntingdon Life Sciences 

Study No. 02-5449 

12. REGULATORY REFERENCES 

12.1. TESTING GUIDELINES 

Page 168 

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Final Protocol 

This study is designed to meet or exceed the requirements of: 

The Organization for Economic Cooperation and Development (OECD) 

Guidelines: Acute Inhalation (12 May 1981 ), Part 403. 

Health Effects Test Guidelines, Office of Prevention, Pesticides and Toxic 

Substances, U.S. Environmental Protection Agency (OPPTS), August, 

1998; OPPTS 870.1300, Acute Inhalation Toxicity. 

12.2. GOOD LABORATORY PRACTICES 

This study will be conducted in compliance with: 

OECD Principles of Good Laboratory Practices ENV/MC/CHEM (98)17 

EPA Good Laboratory Practices as set forth in 40 CFR Part 792 (TSCA) 

12.3. ANIMAL WELFARE ACT COMPLIANCE 

This study will comply with all appropriate parts of the Animal Welfare 

Act Regulations: 9 CFR Parts 1 and 2 Final Rules, Federal Register, 

Volume 54, No. 168, August 31, 1989, pp. 36112-36163 effective 

October 30, 1989 and 9 CFR Part 3 Animal Welfare Standards; Final 

Rule, Federal Register, Volume 56, No. 32, February 15, 1991, pp. 6426-

6505 effective March 18, 1991. The Sponsor should make particular 

note ofthe following: 

1. The Sponsor's signature on this protocol documents that, for the 

study described, there are no generally accepted non-animal 

alternatives and the study does not unnecessarily duplicate previous 

experiments in regard to species, test substance, route of 

administration or duration of treatment. 

2. All procedures used in this study have been designed to avoid 

discomfort, distress and pain to the animals. All methods are 

described in this study protocol or written laboratory standard 

operating procedures. 

3. Any procedures outlined in this study which cause more than 

momentary or slight pain or distress to the animals will be performed 

with appropriate sedatives, analgesics or anesthetics unless the 

withholding of these agents is justified for scientific reasons, in 
Huntingdon Life Sciences 

Study No. 02-5449 

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Final Protocol 

12.4. 

writing by the Sponsor and the Study Director, in which case the 

procedure will continue for the minimum time necessary. 

Documentation of the justification for withholding treatment for pain 

or distress and IACUC approval of the procedures will be made prior 

to study initiation on the IACUC Protocol Review Form. 

4. Animals experiencing more than momentary or slight pain or distress 

due to test substance or injury or illness may be euthanized by the 

Testing Facility's veterinary staff to relieve pain after consultation 

with the Study Director and Sponsor. However, in emergency 

situations, the veterinary staff is authorized to administer emergency 

treatment as necessary. Any subsequent treatment or euthanasia will 

be administered after consultation with the Study Director. The 

Sponsor will be advised by the Study Director of all emergency 

situations in as timely a manner as possible. 

5. Methods of euthanasia used during this study are in conformance 

with the above-referenced regulations. 

INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE 

(IACUC) 

The IACUC Protocol Review Subcommittee has reviewed this protocol 

and found it to be in compliance with all appropriate regulations. 

13. . QUALITY ASSURANCE MONITORING 

The Huntingdon Life Sciences Quality Assurance Unit will monitor the facilities, 

equipment, personnel, methods, practices, records and controls used in this study 

to assure that they are in conformance with this protocol, company SOPs, and the 

appropriate Good Laboratory Practice requirements. 

14. ALTERATION IN STUDY DESIGN 

Alterations of this protocol may be made as the study progresses. No changes in 

the protocol will be made without the consent of the Sponsor. In the event that 

the Sponsor authorizes a protocol change verbally, such changes will be honored 

by the Testing Facility and will be followed by a written verification. All protocol 

modifications will be signed by the Study Director and a Sponsor representative. 

Any modifications potentially affecting animal welfare will also be signed by 2 

members of the Institutional Animal Care and Use Committee prior to the 

modification's implementation. 
Huntingdon Life Sciences 

Study No. 02-5449 

Protocol Amendment No. 1 

Page 170 

Page 1 ofl 

Final Protocol 

Study Title: 2-Bronio-3,3,3-Trifluoropropene, 1-Bromo-3,3,3-Trifluoropropene and 

1-Bromo-2-Trifluoromethyl-3,3,3-Trifluoropropene: An Acute ( 4-Hour) Inhalation 

Toxicity Study in the Rat via Nose-Only Exposure 

Changes 

1. Page 4, Administration of Test Substances: 

Revise: ...... The flow-past nose-only exposure chambers will be operated at a 

minimum flow rate of 6 & liters per minute~ .... 

2. Page 5, Summary of Chamber Activity: 

Revise: The minimum frequency of chamber activity is summarized below: 

Activity 

Particle Size 

3. Page 10, Tissue Preservation: 

Frequency !Exposure 

4-!--X 

Revise: All weighed organs as listed in Section 8.4 will be preserved in 10% neutral 

buffered formalin. The lungs will be infused with 10% neutral buffered formalin to 

ensure fixation and preserved for all animals. The testes will be fixed in modified 

Davidson's solution for at least 48 hours prior to permanent storage in NBF .. :" 

Reasons for Changes 

1. The total flow rate was reduced to reduce the consumption of the test substance . 

. 2. Error in original protocol. 

3. Omission in original protocol. 

Additional Cost Required: No . 

Amendment approved by: 

Gary M. Hoffin:t B.A., D.A.B.T. 

s~ 

Bradford F. Colton 

Sponsor Representative 

3/oslo3 

I I 

Date 
(~ . . . . 

Huntingdon Life Sciences 02-5449 Page 171 

Final Report 

Testing Facility Personnel Appendix D 

TITLE/DEPARTMENT 

SENIOR VICE PRESIDENT, 

SAFETY ASSESSMENT 

SENIOR DIRECTOR, SAFETY 

ASSESSMENT 

DIRECTOR, TOXICOLOGY 

OPERATIONS 

DIRECTOR, QUALITY ASSURANCE 

STUDY DIRECTOR 

PATHOLOGIST 

STUDY MONITOR 

VETERINARIAN 

MANAGER/SUPERVISOR 

Inhalation Toxicology 

Histology 

NAME/DEGREE 

Sylvie J. Gosselin, D.V.M., Ph.D., 

Diplomate A.C.V.P. 

CarolS. Auletta, M.B.A., D.A.B.T., 

R.A.C. 

lan Vanterpool, F.I.A.T. 

Nicki S. Iacono, B.S. 

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

Henry F. Bolte, D.V.M., Ph.D. 

Tara A. Gankiewicz, B.A. 

Teresa S. Kusznir, V.M.D. 

Robert Faust, B.S. 

Janet Kusisto, B.S., H.T. (ASCP) 
Huntingdon Life Sciences 02-5449 

Report Amendments 

There are no amendments for this report at this time. 

Page 172 

Final Report 

Appendix E