Page 1 of 221

Working for a better future

2-bromo-3,3,3-trifluoropropene: Dose Range Finding Study by 

Inhalation Administration to Rats for 2 Weeks

HLS study number: WAG0013

Version ID: Final

Issue date: 21 August 2012
Huntingdon Life Sciences

WAG0013

2

Details of Sponsor and Test Facilities

Sponsor American Pacific Corporation

3883 Howard Hughes Parkway

Suite 700

Las Vegas

Nevada 89169

USA

Test facility Huntingdon Life Sciences

Primary location Huntingdon Research Centre

Woolley Road

Alconbury

Huntingdon

Cambridgeshire

PE28 4HS

UK

Test facility Huntingdon Life Sciences

Histology Eye Research Centre

Eye

Suffolk

IP23 7PX

UK
Huntingdon Life Sciences

WAG0013

3

Table of Contents

Details of Sponsor and Test Facilities ................................................................................... 2

Compliance with Good Laboratory Practice .......................................................................... 6

Quality Assurance Statement................................................................................................ 7

Contributing Scientists .......................................................................................................... 9

Summary ............................................................................................................................ 10

1. Introduction ................................................................................................................ 12

1.1 Objective......................................................................................................... 12

1.2 Regulatory compliance ................................................................................... 12

1.3 Test system .................................................................................................... 12

1.4 Route of administration................................................................................... 12

1.5 Treatment groups and doses .......................................................................... 12

2. Experimental Procedure............................................................................................. 13

2.1 Study schedule and structure.......................................................................... 13

2.1.1 Duration of treatment........................................................................... 13

2.1.2 Time schedule..................................................................................... 13

2.1.3 Identity of treatment groups................................................................. 13

2.2 Test substance blend...................................................................................... 14

2.2.1 Test substance.................................................................................... 14

2.3 Animal management....................................................................................... 15

2.3.1 Animal supply, acclimatisation and allocation...................................... 15

2.3.2 Animal housing, diet and water supply ................................................ 15

2.3.3 Administration ..................................................................................... 16

2.4 Serial observations ......................................................................................... 16

2.4.1 Clinical observations ........................................................................... 17

2.4.2 Detailed physical examination and arena observations ....................... 17

2.4.3 Sensory reactivity and grip strength .................................................... 17

2.4.4 Motor activity....................................................................................... 18

2.4.5 Bodyweight ......................................................................................... 19

2.4.6 Food consumption............................................................................... 19

2.5 Necropsy and Histology.................................................................................. 19

2.5.1 Method of kill....................................................................................... 19

2.5.2 Macroscopic pathology........................................................................ 19

2.5.3 Organ weights..................................................................................... 19

2.5.4 Fixation ............................................................................................... 20

2.5.5 Histology ............................................................................................. 21

2.6 Pathology........................................................................................................ 21

2.6.1 Light microscopy ................................................................................. 21

2.7 Computer systems.......................................................................................... 23

2.8 Data treatment................................................................................................ 23

2.8.1 Definition of “Day” ............................................................................... 24

2.8.2 Signs................................................................................................... 24

2.8.3 Bodyweight ......................................................................................... 24

2.8.4 Pathology............................................................................................ 24

2.8.5 Statistical analysis............................................................................... 24

2.9 Quality assurance and archiving procedures .................................................. 25

2.9.1 Quality assurance ............................................................................... 25

2.9.2 Archives .............................................................................................. 25
Huntingdon Life Sciences

WAG0013

4

2.10 Deviations from protocol ................................................................................. 25

3. Results....................................................................................................................... 26

3.1 Chamber atmosphere conditions .................................................................... 26

3.2 Signs associated with dosing.......................................................................... 26

3.3 Detailed physical examination and arena observations................................... 26

3.4 Sensory reactivity and grip strength................................................................ 26

3.5 Motor activity .................................................................................................. 27

3.6 Bodyweight..................................................................................................... 27

3.7 Food consumption .......................................................................................... 27

3.8 Organ weights................................................................................................. 27

3.9 Macropathology .............................................................................................. 28

3.10 Histopathology................................................................................................ 28

4. Discussion.................................................................................................................. 34

5. Conclusion ................................................................................................................. 36

6. References................................................................................................................. 37

List of Figures

Figure 1 Motor activity................................................................................................38

Figure 2 Bodyweight ..................................................................................................40

List of Tables

Table 1 Sensory reactivity and grip strength .............................................................41

Table 2 Motor activity................................................................................................42

Table 3 Bodyweight ..................................................................................................44

Table 4 Organ weights..............................................................................................48

Table 5 Macropathology ...........................................................................................50

Table 6 Histopathology .............................................................................................52

List of Appendices

Appendix 1 Clinical signs................................................................................................54

Appendix 2 Signs associated with dosing .......................................................................60

Appendix 3 Sensory reactivity and grip strength ...........................................................110

Appendix 4 Motor activity..............................................................................................112

Appendix 5 Bodyweight ................................................................................................120

Appendix 6 Food consumption .....................................................................................124

Appendix 7 Organ weights............................................................................................128

Appendix 8 Macropathology and histopathology...........................................................130
Huntingdon Life Sciences

WAG0013

5

List of Annexes

Annex 1 Administration of 2-bromo-3,3,3-trifluoropropene to rats ............................188

Annex 2 Pathology report.........................................................................................206

Annex 3 Certificate of analysis.................................................................................217

Annex 4 GLP compliance statements ......................................................................219

Huntingdon Life Sciences

WAG0013

7

Quality Assurance Statement

2-bromo-3,3,3-trifluoropropene: Dose range finding study by inhalation 

administration to rats for 2 weeks

The following inspections and audits have been carried out in relation to this study:

Study Phase

Protocol Audit

Protocol Amendment No 1

Protocol Amendment No 2

Protocol Amendment No 3

Protocol Amendment No 4

Date(s) of Inspection

25 Oct 2011

16 Nov 2011

16 Nov 2011

25 Nov 2011

10 Feb 2012

Date of Reporting to Study 

Director and Management

25 Oct 2011

16 Nov 2011

16 Nov 2011

25 Nov 2011

10 Feb 2012

Report Audit

Report Audit

25 Jan 2012-30 Jan 2012

22 Mar 2012-27 Mar 2012

30 Jan 2012

27 Mar 2012

Process based inspections: At or about the time this study was in progress inspections of

procedures employed on this type of study were carried out. These were conducted and 

reported to appropriate Company Management as indicated below:

Process Based Inspections Date(s) of Inspection Date of Reporting to 

Management

Clinical signs 

examination/palpations including 

arena observations (rodents)

17 Oct 2011 17 Oct 2011

Study management and conduct 25 Oct 2011 25 Oct 2011

Bodyweights and food consumption 31 Oct 2011 31 Oct 2011

Motor activity testing on RAMS 

(including home/in the 

hand/standard arena/manipulations)

01 Dec 2011 01 Dec 2011

Generation of Test Atmospheres and 

Exposure/post exposure signs, 

atmosphere sampling

13 Dec 2011 13 Dec 2011

Test Item Control and disposition 

(including tracking and canister 

packing)

14 Dec 2011 14 Dec 2011

Terminal procedures 14Dec 2011 14 Dec 2011

Huntingdon Life Sciences

WAG0013

9

Contributing Scientists

2-bromo-3,3,3-trifluoropropene: Dose range finding study by inhalation 

administration to rats for 2 weeks

Study management

Paul J Horrell BSc (Hons) IDT

Study Director

William N Hooks MIBMS BSc (Hons)

Senior Toxicologist

Inhalation analysis

Caroline Finucane PhD

Head of Formulation & Inhalation Analysis

Aerosol technology

Simon A Moore BSc (Hons) PhD MRSC

Head of Aerosol Technology

Sensory reactivity, grip strength and motor activity

Martin J Collier BSc

Behavioural Scientist

Pathology

David Bell, BSc (Hons) BVMS MRCVS

Pathologist
Huntingdon Life Sciences

WAG0013

10

Summary

The objective of this study was to assess the local and systemic toxic potential of

2-bromo-3,3,3-trifluoropropene, a fire extinguishing agent, when administered by inhalation

to Crl:CD(SD) rats for 6 hours a day, 5 days a week, for 2 weeks.

Three groups, each comprising three male and three female rats received

2-bromo-3,3,3-trifluoropropene at target exposure levels of 5000, 10000 or 20000 ppm. A 

similarly constituted Control group received clean air only, at the same volume-dose.

During the study, clinical condition, detailed physical and arena observations, sensory 

reactivity, grip strength, motor activity, bodyweight, food consumption, organ weight, 

macropathology and histopathology investigations were undertaken.

Results

The achieved aerosol concentrations were 4721, 8901 and 19098 ppm (94, 89 and 95% of 

target) for Groups 2-4 respectively.

There were no unscheduled deaths at any of the dose levels.

Treatment-related, but transient, clinical signs, related to inhalation of an irritant material or 

potential CNS effects, were evident during and after exposure. Bodyweight loses of 43g and 

3g were evident for males and females exposed to 19098 ppm respectively over the treatment 

period. Reduced bodyweight gain was evident for males exposed to 8901 or 4721 ppm 

(0.02X and 0.23X control respectively). Total food consumption was reduced for all treated 

groups (0.66X to 0.89X Control).

At necropsy, a reduction in the size of the thymus and depressions or roughening of the 

stomach were evident in animals exposed to 19098 ppm. All other organs were within the 

normal size and weight ranges.

Histopathological changes associated with to treatment with 2-bromo-3,3,3-trifluoropropene 

were seen in the nasal turbinates, larynx and trachea. Overall, the most marked changes were 

in the nasal cavity, with minimal to slight changes in the larynx and trachea. 

In the region of the olfactory epithelium of the nasal turbinates, fibrosis, with or without 

associated cartilage formation, atrophy of the Bowman’s glands, atrophy of olfactory nerve 

fibres and epithelial erosion were seen in animals of all test article groups. Necrosis of the 

olfactory epithelium was seen in animals exposed to 8901 or 19098 ppm. Fusion of dorsal 

turbinates with associated epithelial diverticula formation was seen in animals of all test 

article groups. In the various epithelia of the nasal cavity (olfactory, respiratory and 

transitional) of animals of all test article groups, metaplasia and combinations of 

disorganisation, atrophy, degeneration and regeneration were evident. In the nasolacrimal 

ducts, inflammation was seen in animals of all test article groups.

In general, the changes seen in the nasal turbinates of animals exposed to 8901 or 19098 ppm 

were of a similar severity and incidence and of a higher severity than those seen for animals 

exposed to 4721 ppm. In consideration of the epithelial changes the olfactory epithelium was 

the most extensively affected at all three exposure levels.
Huntingdon Life Sciences

WAG0013

11

In the larynx, epithelial metaplasia, hyperplasia and keratinisation, of a similar incidence and 

degree, were seen in animals of all test article groups. Inflammation was seen in one male 

and one female exposed to 19098 ppm and one male exposed to 8901 ppm.

In the trachea, epithelial degeneration/regeneration was seen in male and female animals 

exposed to 19098 ppm and one female exposed to 8901 ppm. Inflammatory cell infiltrate in 

the lamina propria was seen at a higher incidence and/or degree in animals exposed to 

19098 ppm, compared with control animals.

Conclusion

Histopathological changes related to treatment were seen in the nasal turbinates, larynx and 

trachea. Although the majority of these changes were limited to the 8901 or 19098 ppm 

exposure groups, some changes in the nasal turbinates were evident at 4721 ppm. Therefore, 

a no observed adverse effect level (NOAEL) could not be established.
Huntingdon Life Sciences

WAG0013

12

1. Introduction

1.1 Objective

The objective of this study was to assess the local and systemic toxic potential of

2-bromo-3,3,3-trifluoropropene, a fire extinguishing agent, when administered by inhalation

to rats for 2 weeks to select suitable exposure levels for a subsequent 13 week toxicity study.

1.2 Regulatory compliance

Due to the preliminary nature of this study no specific regulations or guidelines are 

applicable.

1.3 Test system

The rat was chosen as the test species because of its acceptance as a predictor of toxic change 

in man and the requirement for a rodent species by regulatory agencies. The Crl:CD (SD)

strain was used because of the historical control data available in this laboratory.

1.4 Route of administration

The inhalation route of administration was chosen to simulate the conditions of potential 

human exposure.

1.5 Treatment groups and doses

In an acute inhalation toxicity study (Laboratory Project Number 02-5449), 0/10 rats died at 

5173 ppm and 10/10 rats died at 26580 ppm. There were no intermediate groups. Clinical 

signs at 5173 ppm comprising respiratory sounds persisted for up to 8 days post-exposure in a 

small number of rats (maximum 4). The rats were weighed on Days 7 and 14 post exposure, 

the weight gain over this period was considered to be normal and unaffected by exposure. 

Lung weights were normal.

Based on these findings, a target exposure level of 5000 ppm, for the low dose group, was 

considered likely to elicit a mild to moderate clinical response with no mortality.

The starts of exposures for treatment groups were staggered (i.e. the groups were exposed 

sequentially) by 7 days to allow assessment of the response of animals. Thus the subsequent 

target exposure levels of 10000 and 20000 ppm, selected for the intermediate and high dose 

groups, were based on the response of the animals at lower dose levels.
Huntingdon Life Sciences

WAG0013

13

2. Experimental Procedure

2.1 Study schedule and structure

2.1.1 Duration of treatment

The test substance, 2-bromo-3,3,3-trifluoropropene, was administered over a period of 

2 consecutive weeks. The necropsy procedures were completed in 1 day for each group 

(staggered over 3 separate days), during which time treatment continued, and serial 

observations were recorded at appropriate intervals. The duration of treatment is reported as 

2 weeks.

2.1.2 Time schedule

Study initiation:

(Protocol signed by Study Director)

24 October 2011

Experimental start date:

(Animal arrival)

27 October 2011

Treatment commenced:

Group 2

Group 3

Groups 1 and 4

11 November 2011

18 November 2011

25 November 2011

Necropsy completed:

Group 2

Group 3

Groups 1 and 4

25 November 2011

02 December 2011

09 December 2011

Experimental completion date:

(pathology)

9 March 2012

Study completion: 21 August 2012

2.1.3 Identity of treatment groups

The study consisted of one Control and three treated groups of rats, identified as follows:

Group Treatment

Target 

exposure level

(ppm)

No. of animals Animal numbers Cage numbers

Male Female Male Female Male Female

1 Control 0 3 3 1-3 13-15 1 5

2 2-bromo-3,3,3-

trifluoropropene

5000 3 3 4-6 16-18 2 6

3 2-bromo-3,3,3-

trifluoropropene

10000 3 3 7-9 19-21 3 7

4 2-bromo-3,3,3-

trifluoropropene

20000 3 3 10-12 22-24 4 8
Huntingdon Life Sciences

WAG0013

14

2.2 Test substance blend

2.2.1 Test substance

Information supplied by the Sponsor regarding the test substance is contained in the test 

substance data sheet, which is retained in study records, and the Certificate of Analysis, 

which is presented in Annex 3.

The following information is given in summary:

Identification: 2-bromo-3,3,3-trifluoropropene (stabilized)

Usage: Fire extinguishing agent

Appearance: Liquid at below 33C

Clear and may be colourless to light yellow tint

Storage conditions: At ambient temperature

Supplier: Sponsor

Batch number: KgF080

Date of receipt: 12 September 2011

Quantity received: 2 × 150 kg cylinders

Reassay date: 1 August 2012

Purity: 99.9% excluding stabilizers

Stabilisers:

The Sponsor was responsible for the characterisation of the test substance and the 

documentation of the methods of synthesis, fabrication or derivation and stability.
Huntingdon Life Sciences

WAG0013

15

2.3 Animal management

2.3.1 Animal supply, acclimatisation and allocation

Crl:CD (SD) rats (a total of fifteen male and fifteen female) were received from Charles 

River (UK) Ltd. The rats were ordered at 49 to 56 days of age and within a weight range of 

290-330 g for males and 202-233 g for females.

On arrival, the animals were removed from the transit boxes and allocated to study cages. 

Using the sequence of cages in the battery, one animal at a time was placed in each cage with 

the procedure being repeated until each cage held the appropriate number of animals. Each 

sex was allocated separately.

The cages constituting each group were blocked together by sex and the groups were 

dispersed in batteries so that possible environmental influences arising from their spatial 

distribution were equilibrated, as far as was practicable.

Each animal was assigned a number and identified uniquely within the study by a tail tattoo. 

Each cage label was colour-coded according to group and was numbered uniquely with cage 

and study number, as well as the identity of the occupants.

The animals were allowed to acclimatise to the conditions described below for at least 

15 days before treatment commenced. For those animals selected for this study, their age at 

the start of treatment was 64 to 85 days and their bodyweights were in the range of 396 to 

501 g for males and 247 to 316 g for females.

The spare animals were removed from the study room after treatment commenced.

2.3.2 Animal housing, diet and water supply

Animals were housed inside a restricted access rodent facility (Building Y14, Room 011). 

Sensory reactivity, grip strength and motor activity tests were performed in Room 010. The 

facility was designed and operated to minimise the entry of external biological and chemical

agents and to minimise the transference of such agents between rooms. Before the study the 

room was cleaned and disinfected.

Each animal room was supplied with filtered fresh air, which was passed to atmosphere and 

not re-circulated. The temperature and relative humidity controls were maintained within the 

range of 19 to 23°C and 40 to 70% respectively and monitored continuously. There were no 

deviations from these ranges. Artificial lighting was controlled to give a cycle of 12 hours 

continuous light and 12 hours continuous dark per 24 hours.

Alarms were activated if there was any failure of the ventilation system, or temperature limits 

were exceeded. A stand-by electricity supply was available to be automatically brought into 

operation should the public supply fail.

The animals were housed three of one sex per cage. The cages were made of a polycarbonate 

body with a stainless steel mesh lid. Wood based material was used as bedding and was 

sterilised by autoclaving and changed at appropriate intervals each week. Cages, food 

hoppers and water bottles were changed at appropriate intervals.
Huntingdon Life Sciences

WAG0013

16

The animals were allowed free access to a standard rodent diet (Rat and Mouse No. 1 

Maintenance Diet). This diet contained no added antibiotic or other chemotherapeutic or 

prophylactic agent.

Potable water taken from the public supply was freely available via polycarbonate bottles 

fitted with sipper tubes.

Each cage of animals was provided with an Aspen chew block for environmental enrichment. 

Chew blocks were provided throughout the study and were replaced when necessary. Each 

cage of animals was provided with a plastic shelter for environmental enrichment, which was 

replaced at the same time as the cages.

Each batch of diet was analysed routinely by the supplier for various nutritional components 

and chemical and microbiological contaminants. Supplier’s analytical certificates were 

scrutinised and approved before any batch of diet was released for use. The quality of the 

water supply is governed by regulations published by the Department for Environment, Food 

and Rural Affairs. Certificates of analysis were received routinely from the water supplier

and the suppliers of the bedding and Aspen chew blocks. Since the results of these various 

analyses did not provide evidence of contamination that might have prejudiced the study, 

they are not presented.

No other specific contaminants that were likely to have been present in the bedding, chew 

blocks, diet or water were analysed, as none that may have interfered with or prejudiced the 

outcome of the study was known.

2.3.3 Administration

Details of administration and analysis of the test atmospheres together with the results 

obtained are presented in Annex 1.

Each exposure system consisted of a Glass vaporiser and a whole body exposure chamber. 

Different doses of 2-bromo-3,3,3-trifluoropropene were achieved by varying the 

concentrations of the test article in the system whilst keeping the duration of exposure 

constant. Group 1 was exposed to clean air only at a target concentration the same as that 

specified for Group 4. Groups 2, 3 and 4 were exposed to 2-bromo-3,3,3-trifluoropropene, 

6 hours/day for 5 days each week (Monday to Friday) for 2 weeks.

2.4 Serial observations

Dated and signed records of all activities relating to the day by day running and maintenance 

of the study within the animal unit as well as to the group observations and examinations 

outlined in this experimental procedure were recorded in the Study Day Book. In addition, 

observations relating to individual animals made throughout the day were recorded.

All observations described below were performed in cage number sequence except where 

otherwise indicated.
Huntingdon Life Sciences

WAG0013

17

2.4.1 Clinical observations

Animals were inspected visually at least twice daily for evidence of ill-health or reaction to 

treatment. Cages were inspected daily for evidence of ill-health amongst the occupant(s). 

Any deviation from normal was recorded at the time in respect of nature and severity, date 

and time of onset, duration and progress of the observed condition, as appropriate.

Daily, on exposure days throughout the treatment period, detailed observations were 

performed immediately before dosing and between one and two hours after completion of 

dosing of all groups. Further observation was performed during exposure and as late as 

possible in the working day during the first week of treatment.

During the acclimatisation period, observations of the animals and their cages were recorded 

once per day.

2.4.2 Detailed physical examination and arena observations

Before treatment commenced and on Day 10 of treatment, detailed physical examination and 

arena observations were performed on each animal. On each occasion, the examinations 

were performed at approximately the same time of day (before dosing during the treatment 

period).

After removal from the home cage, animals were assessed for physical condition and 

behaviour during handling and after being placed in a standard arena. Any deviation from 

normal was recorded with respect to the nature and, where appropriate, degree of severity. 

Particular attention was paid to possible signs of neurotoxicity, such as convulsions, tremor 

and abnormalities of gait or behaviour.

2.4.3 Sensory reactivity and grip strength

Sensory reactivity and grip strength assessments were performed (before dosing) on all

animals on Day 10 of treatment. 

The following measurements, reflexes and responses were recorded:

Approach response

A blunt probe was brought towards the animal’s head until it was close to the animal’s nose 

(but not touching the vibrissae). The animal’s reaction was recorded as:

1 No reaction or ignores probe

2 Normal awareness and reaction e.g. approaches and/or sniffs probe

3 Abnormally fearful or aggressive reaction
Huntingdon Life Sciences

WAG0013

18

Touch response

The animal was stroked gently on the nape of the neck with a blunt probe and the reaction 

recorded as:

1 No reaction or ignores probe

2 Normal awareness and reaction e.g. turns towards or moves away

3 Abnormally fearful or aggressive reaction

Auditory startle reflex

The animal’s response to a sudden loud noise was assessed and scored as:

1 No response

2 Weak response e.g. ear twitch only

3 Normal response e.g. obvious flinch or startle

4 Exaggerated response e.g. all feet off floor

Tail pinch response

The animal’s tail was pinched sharply with forceps approximately one third from the tip and 

the response graded as:

1 No response

2 Weak response e.g. turns around slowly or weak vocalisation without 

moving away

3 Normal response e.g. jumps forward or turns around sharply, usually with

vocalisation

4 Exaggerated response e.g. excessive vocalisation, body movement or 

aggression

Grip strength

Forelimb and hindlimb grip strength was measured using Mecmesin Force Indicators. Three 

trials were performed.

At any point during the observations, additional comments were made as free text where 

considered appropriate.

2.4.4 Motor activity

On Day 10 of treatment (before dosing), the motor activity of each animal was measured 

using a Rodent Activity Monitoring System (Version 2.0.3), with hardware supplied by 

Pearson Technical Services and software developed and maintained by Huntingdon Life 

Sciences. 

Animals were tested individually in clear polycarbonate cages and motor activity was 

measured by counting infra-red beam breaks over ten 6-minute intervals (one hour total). 

Ten beams were set at two height levels (five low and five high) to detect cage floor and 

rearing activity respectively.
Huntingdon Life Sciences

WAG0013

19

2.4.5 Bodyweight

The weight of each rat was recorded daily from Day -5, daily throughout the treatment, and 

before necropsy.

2.4.6 Food consumption

The weight of food supplied to each cage, that remaining and an estimate of any spilled was 

recorded daily from Day -5, and each day throughout the treatment period.

2.5 Necropsy and Histology

2.5.1 Method of kill

Animals were killed by intra-peritoneal sodium pentobarbitone, followed by exsanguination.

2.5.2 Macroscopic pathology

All animals were subject to a detailed necropsy.

After a review of the history of each animal, a full macroscopic examination of the tissues 

was performed. All external features and orifices were examined visually. The cranial roof 

was removed to allow observation of the brain, pituitary gland and cranial nerves. After 

ventral mid-line incision, the neck and associated tissues and the thoracic, abdominal and 

pelvic cavities and their viscera were exposed and examined in situ. Any abnormal position, 

morphology or interaction was recorded.

The requisite organs were weighed and external and cut surfaces of the organs and tissues 

were examined as appropriate. Any abnormality in the appearance or size of any organ and 

tissue was recorded and the required tissue samples preserved in appropriate fixative.

The retained tissues were checked before disposal of the carcass.

2.5.3 Organ weights

The following organs, taken from each animal killed after 2 weeks of treatment, were 

dissected free of adjacent fat and other contiguous tissue and the weights recorded:

Brain Liver

Heart Lungs with mainstem bronchi

Kidneys Spleen

Bilateral organs were weighed together. Organ weights were also adjusted for terminal 

bodyweight, using the weight recorded before necropsy.
Huntingdon Life Sciences

WAG0013

20

2.5.4 Fixation

Testes were fixed in modified Davidson's fluid and eyes were fixed in Davidson's fluid. 

Samples (or the whole) of the other tissues listed below from all animals were preserved in 

10% neutral buffered formalin:

Adrenals Optic nerves

Aorta - thoracic Ovaries

Brain Pancreas

Caecum Pituitary

Colon Prostate

Duodenum Rectum

Epididymides Salivary glands - submandibular

Eyes - sublingual

Femurs Sciatic nerves

Harderian glands Seminal vesicles

Head Skeletal muscle

Heart Skin with mammary glands

Ileum Spinal cord

Jejunum Spleen

Kidneys Sternum

Lachrymal glands Stomach

Larynx* Testes

Liver Thymus

Lungs including bronchi* Thyroid with parathyroids

Lymph nodes - mandibular Tongue

- mesenteric Trachea*

- tracheo-bronchial Urinary bladder

Nasal turbinates* Uterus and cervix

Oesophagus Vagina

* Tissues examined microscopically

Samples of any abnormal tissues were also retained. In those cases where a lesion was not 

clearly delineated, contiguous tissue was fixed with the grossly affected region and sectioned 

as appropriate.

Samples of the head (including nasal cavity, paranasal sinuses and nasopharynx) and the 

remaining femur, salivary gland, sciatic nerve and skeletal muscle were not examined 

histologically, but are retained against any future requirement for microscopic examination.
Huntingdon Life Sciences

WAG0013

21

2.5.5 Histology

For those animals specified in the Pathology section, the relevant tissues were subject to 

histological processing.

Tissue samples were dehydrated, embedded in paraffin wax, sectioned at approximately four 

to five micron thickness and stained with haematoxylin and eosin.

Those tissues subject to histological processing included the following regions:

Larynx - 5 sections

Lungs - section from all major lobes, to include bronchi

Nasal Turbinates - 3 sections

Trachea - including bifurcation

2.6 Pathology

2.6.1 Light microscopy

Microscopic examination was performed as follows:

Larynx, lungs, nasal turbinates and trachea were examined for all animals of 

Groups 1 (Control) and 4 (20000 ppm) sacrificed on completion of the scheduled 

treatment period.

Larynx, nasal turbinates and trachea were examined for all animals of Groups 2 

(5000 ppm) and 3 (10000 ppm) sacrificed on completion of the scheduled treatment 

period.

Tissues reported at macroscopic examination as being grossly abnormal were 

examined in line with current practice.

Findings were either reported as "present" or assigned a severity grade. In the latter case one 

of the following five grades was used - minimal, slight, moderate, marked or severe. 

The descriptions listed below are an attempt to define the grades in general terms for all 

microscopic findings reported in a study and are not specific for a particular tissue. They are 

more strictly applicable to some types of lesion than to others, and there may therefore be a 

few lesions for which these definitions are not completely appropriate.
Huntingdon Life Sciences

WAG0013

22

Thresholds for the grading of lesions for certain tissues may vary with type of study and also 

may vary, generally for all tissues or the majority of tissues, with the duration of the study.

In increasing order of degree/severity.

Minimal The lowest grade of a lesion which is discernible/distinguishable from 

normal and is often subtle and/or involving a very small area of the 

organ.

Slight Clearly discernible/distinguishable from normal with more extensive or 

more apparent changes than minimal.

Moderate Generally involves a significant proportion of the affected organ or 

tissue.

Marked Involves the majority of the affected organ or tissue.

Severe Involves all or almost all of an affected organ or tissue.

A reviewing pathologist undertook a peer review of the microscopic findings.
Huntingdon Life Sciences

WAG0013

23

2.7 Computer systems

The computer systems with version numbers that were used on this study to acquire and 

quantify data include:

Liberate Release 2 in-house system used for reporting clinical signs, dosing signs, 

bodyweights, food consumption, organ weights, macropathology and 

histopathology.

RAMS Rodent Activity Monitoring System (version 2.0) used for activity 

monitoring.

Responder - X (version 5.11) used for startle response.

Sample Registry System (version 1.0) used for Formulation and Inhalation Analysis 

data.

Waters Empower 2, Build 2154, Feature Release 4, Service Pack D and used for 

Formulation and Inhalation Analysis.

Xybion Path/Tox System (version 4.2.2) used for clinical signs, dosing signs, 

bodyweights, food consumption, organ weights, macropathology and 

histopathology.

Xybion Pristima (version 6.2) used for Pharmacy test substance management.

2.8 Data treatment

This report contains serial observations pertaining to all weeks of treatment completed, 

together with signs data collected during the necropsy period. The only serial observations 

relating to the acclimatisation period included in this report relate to the detailed physical 

examination and arena observations, Week -1 bodyweights and food consumption.

Summary statistics (e.g. means and standard deviations) presented in this report were 

calculated from computer-stored individual raw data. The summary statistics and the 

individual data were stored in the computer to a certain number of decimal places, different 

for each parameter. For presentation purposes, however, they were usually rounded to fewer 

places. It is, therefore, not generally possible to reproduce the presented means and standard 

deviations exactly using the presented individual data.

Throughout the report the following abbreviations are used:

M Male

F Female

N Number of animals/cages examined

SD Standard deviation

The death codes in the appendices have the following meaning:

T Scheduled kill
Huntingdon Life Sciences

WAG0013

24

2.8.1 Definition of “Day”

The first day of treatment started at midnight prior to treatment commencing and ended at 

midnight on the day following. Subsequent experimental days of treatment were of the same 

duration.

2.8.2 Signs

Signs are considered in two parts: observations made in association with treatment, classified 

as “dosing signs” and extended changes in condition, classified as “clinical signs”. Both are 

presented for each animal that showed signs, providing detail of type of sign, day of

occurrence and information on the duration of the sign applicable.

2.8.3 Bodyweight

Group mean weight changes were calculated from the weight changes of individual animals.

2.8.4 Pathology

The abbreviations used in the Appendix have the following meanings:

Ln/LN Lymph nodes

Bifurc. Bifurcation

Tissues which could not be examined are specified in the Appendix. The absence of a 

comment for a tissue scheduled for examination therefore indicates that the tissue was 

considered to be normal. In all tabular presentations of data the tissues specified in the 

protocol for histopathological examination precede other tissues.

2.8.5 Statistical analysis

The small sample size precludes meaningful statistical evaluation.
Huntingdon Life Sciences

WAG0013

25

2.9 Quality assurance and archiving procedures

2.9.1 Quality assurance

Details of the Quality Assurance inspections and audits (undertaken at Huntingdon Life 

Sciences) are presented on the Quality Assurance Statement.

2.9.2 Archives

Following completion of this study all raw data, specimens and samples, except those 

generated or used during any Sponsor’s or supplier’s analysis, were stored in the archives of 

Huntingdon Life Sciences. Types of sample and specimen which are unsuitable, by reason of 

instability, for long term retention and archiving may be disposed of after the periods stated 

in Huntingdon Life Sciences Standard Operating Procedures.

A copy of the final report and all Quality Assurance inspection records will be retained 

indefinitely. All other appropriate specimens and records will be retained for a minimum 

period of one year from the date of issue of the final report. At the end of the one year 

retention period the Sponsor will be contacted and advice sought on the above requirements. 

Under no circumstances will any item be discarded without the Sponsor’s knowledge.

2.10 Deviations from protocol

There were no deviations from protocol.
Huntingdon Life Sciences

WAG0013

26

3. Results

3.1 Chamber atmosphere conditions

Annex 1

The achieved chamber concentrations are summarised as follows:

Group Aerosol concentration (ppm)

Target Study mean

2 5000 4721

3 10000 8901

4 20000 19098

The achieved aerosol concentrations were 94, 89 and 95% of the target concentration for 

Groups 2, 3 and 4, respectively.

3.2 Signs associated with dosing

Appendix 2

There were no unscheduled deaths during the course of the study.

Clinical signs seen during exposure included: underactivity, shallow breathing, piloerection, 

unsteady gait, partially closed eyelids, salivation and chin rubbing. These signs generally 

resolved quickly on cessation of exposure to the test article. The number and prevalence of 

signs increased with dose. 

3.3 Detailed physical examination and arena observations

Appendix 1

Detailed physical examinations during Week 2 of exposures showed underactivity for 3 out 6 

rats and piloerection for 2 out of 6 rats exposed to 19098 ppm. Abnormal gait and reduced 

body temperature were also evident for a single female exposed to 19098 ppm.

3.4 Sensory reactivity and grip strength

Table 1 and Appendix 3

At least one of the males or females exposed to 8901 or 19098 ppm failed to respond during 

approach response, touch response and auditory startle reflex tests, although there was no 

consistent dose-response. However, all animals showed a tail pinch response and sensory 

reactivity responses for males and females exposed to 4721 ppm were unaffected. There 

were no clear treatment-related changes in grip strength values.
Huntingdon Life Sciences

WAG0013

27

3.5 Motor activity

Figure 1, Table 2 and Appendix 4

High beam and low beam motor activity scores (rearing and cage floor activity respectively) 

for males and females exposed to 8901 or 19098 ppm were markedly reduced throughout the 

1-hour recording period. The effect was more marked in females, although a more consistent 

dose-relationship was evident in males. Motor activity scores for females exposed to 

4721 ppm were also reduced but activity for males at this level was considered to be 

unaffected.

3.6 Bodyweight

Figure 2, Table 3 and Appendix 5

Bodyweight loses of 43g and 3g were evident for males and females exposed to 19098 ppm 

respectively over the treatment period. Reduced bodyweight gain was evident for males 

exposed to 8901 or 4721 ppm (0.02X and 0.23X control respectively). Body weight gains for 

females exposed to 8901 or 4721 ppm greater than control (1.68X and 1.35X control 

respectively).

3.7 Food consumption

Appendix 6

Food consumption was reduced over the treatment period for rats exposed to 

2-bromo-3,3,3-trifluoropropene. 

Total food consumption (Days 1 to 14) was 0.87X, 0.89X and 0.67X control for males and 

0.73X, 0.85X and 0.66X of control for females exposed to 4721, 8901 or 19098 ppm 

respectively.

3.8 Organ weights

Table 4 and Appendix 7

There were no test article-related effects on organ weights.
Huntingdon Life Sciences

WAG0013

28

3.9 Macropathology

Table 5 and Appendix 8

The macroscopic examination performed after 2 weeks of treatment revealed the following 

intergroup differences:

Thymus

A reduction in the size of the thymus was noted in animals treated with 19098 ppm.

Summary of findings in the thymus for animals killed after 2 weeks of treatment

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098

Small 0 0 0 3 0 0 0 3

Number of animals examined 3 3 3 3 3 3 3 3

Stomach

Depressions or roughening of the stomach was observed in animals treated with 19098 ppm.

Summary of findings in the stomach for animals killed after 2 weeks of treatment

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098

Forestomach Depression(s) 0 0 0 2 0 0 0 0

Forestomach Roughened 0 0 0 1 0 0 0 2

Number of animals examined 3 3 3 3 3 3 3 3

The nature and incidence of all other findings were consistent with the commonly seen 

background of macroscopic changes in rats at these laboratories.

3.10 Histopathology

Table 6 and Appendix 8

Changes related to treatment with 2-bromo-3,3,3-trifluoropropene were seen in the nasal 

turbinates, larynx and trachea.

Nasal turbinates

In the olfactory epithelium of male and female animals exposed to concentrations of 4721, 

8901 or 19098 ppm, the test article-related findings seen included the following.

Minimal to marked fibrosis with or without associated cartilage formation in the dorsal 

lamina propria.

Minimal to marked atrophy of the Bowman’s glands in the dorsal lamina propria.
Huntingdon Life Sciences

WAG0013

29

Minimal to moderate atrophy of olfactory nerve fibres.

Minimal epithelial necrosis was seen in two male animals exposed to 19098 ppm and one 

female exposed to 8901 ppm.

Minimal erosion of the epithelium.

Minimal to moderate degeneration/atrophy/disorganisation of the epithelium.

Minimal squamous metaplasia of the epithelium.

Minimal or slight respiratory metaplasia of the epithelium with or without associated 

inflammatory cell infiltrate.

In the turbinates, minimal to moderate fusion of dorsal turbinates with associated epithelial 

diverticula formation was seen in male and female animals exposed to 4721, 8901 or 

19098 ppm.

In the respiratory epithelium of male and female animals exposed to 4721, 8901 or 

19098 ppm, the test article-related findings seen included the following.

Minimal or slight disorganisation/degeneration/regeneration of the epithelium.

Minimal squamous metaplasia of the epithelium was only seen in animals exposed to 

19098 ppm.

In the transitional epithelium, minimal or slight disorganisation/regeneration of the 

epithelium was seen in male and female animals exposed to 4721, 8901 or 19098 ppm.

In the nasolacrimal ducts, minimal epithelial degeneration was seen in one female exposed to 

19098 ppm and minimal or slight inflammation was seen in animals exposed to 4721, 8901 or 

19098 ppm. In the airway, minimal or slight debris and inflammatory cells were also seen in 

animals exposed to 4721, 8901 or 19098 ppm.
Huntingdon Life Sciences

WAG0013

30

Summary of treatment related findings in the nasal turbinates for animals killed after 2 weeks of 

treatment

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098

Olfactory epithelium

Fibrosis with or without associated 

cartilage formation in dorsal lamina 

propria

Minimal

Slight

Moderate

Marked

0

0

0

0

2

0

0

0

0

1

2

0

0

1

0

2

0

0

0

0

3

0

0

0

0

0

3

0

0

0

3

0

Total 0 2 3 3 0 3 3 3

Bowman’s gland atrophy in dorsal 

lamina propria

Minimal

Slight

Moderate

Marked

0

0

0

0

2

1

0

0

0

3

0

0

0

0

1

2

0

0

0

0

0

0

3

0

0

0

3

0

0

0

3

0

Total 0 3 3 3 0 3 3 3

Olfactory nerve fibres - atrophy

Minimal

Slight

Moderate

0

0

0

1

2

0

0

3

0

0

3

0

0

0

0

2

1

0

0

2

1

0

2

1

Total 0 3 3 3 0 3 3 3

Necrosis

Minimal 0 0 0 2 0 0 1 0

Total 0 0 0 2 0 0 1 0

Erosion

Minimal 0 1 1 2 0 1 1 1

Total 0 1 1 2 0 1 1 1

Degeneration/atrophy/disorganisation

Minimal

Slight

Moderate

0

0

0

3

0

0

1

1

1

0

2

1

0

0

0

1

1

0

0

3

0

0

1

2

Total 0 3 3 3 0 2 3 3

Squamous metaplasia

Minimal 0 3 2 1 0 3 3 2

Total 0 3 2 1 0 3 3 2

Respiratory metaplasia with or 

without associated inflammatory cell 

infiltrate

Minimal

Slight

0

0

2

0

1

2

1

2

0

0

3

0

3

0

2

0

Total 0 2 3 3 0 3 3 2
Huntingdon Life Sciences

WAG0013

31

Summary of treatment related findings in the nasal turbinates for animals killed after 2 weeks of 

treatment

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098

Turbinates

Fusion of dorsal turbinate with 

associated epithelial diverticula 

formation

Minimal

Slight

Moderate

0

0

0

0

0

0

0

0

3

0

1

2

0

0

0

1

0

0

0

0

3

0

0

3

Total 0 0 3 3 0 1 3 3

Respiratory epithelium

Disorganisation/Degeneration/

Regeneration

Minimal

Slight

0

0

1

0

3

0

3

0

0

0

1

0

1

1

2

1

Total 0 1 3 3 0 1 2 3

Squamous metaplasia

Minimal 0 0 0 3 0 0 0 1

Total 0 0 0 3 0 0 0 1

Transitional epithelium

Disorganisation/Regeneration

Minimal

Slight

0

0

2

0

2

0

2

1

0

0

2

0

2

0

2

1

Total 0 2 2 3 0 2 2 3

Nasolacrimal ducts

Epithelial degeneration

Minimal 0 0 0 0 0 0 0 1

Total 0 0 0 0 0 0 0 1

Inflammation

Minimal

Slight

0

0

1

0

1

0

0

2

0

0

1

0

0

0

2

0

Total 0 1 1 2 0 1 0 2

Airway

Debris and inflammatory cells

Minimal

Slight

0

0

2

0

1

0

0

1

0

0

0.

0

2

0

1

0

Total 0 2 1 1 0 0 2 1

Number of tissues examined 3 3 3 3 3 3 3 3
Huntingdon Life Sciences

WAG0013

32

Larynx

In the ventral larynx, minimal squamous metaplasia was seen in male and female animals 

exposed to 8901 or 19098 ppm and minimal epithelial hyperplasia was seen in one male 

exposed to 19098 ppm.

In the ventrolateral larynx, minimal epithelial keratinisation was seen in male and female 

animals of all groups exposed to 2-bromo-3,3,3-trifluoropropene.

In the arytenoids, minimal or slight squamous metaplasia and minimal epithelial hyperplasia 

were seen in animals of all test article groups.

Minimal or slight inflammation was seen in one male and one female exposed to 19098 ppm 

and one male exposed to 8901 ppm.

Summary of treatment related findings in the larynx for animals killed after 2 weeks of treatment

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098

Squamous metaplasia - ventral

Minimal 0 0 1 2 0 0 3 3

Total 0 0 1 2 0 0 3 3

Epithelial hyperplasia - ventral

Minimal 0 0 0 1 0 0 0 0

Total 0 0 0 1 0 0 0 0

Epithelial keratinisation -

ventrolateral

Minimal 0 2 2 3 0 2 2 3

Total 0 2 2 3 0 2 2 3

Squamous metaplasia - arytenoid

Minimal

Slight

0

0

1

1

3

0

1

1

0

0

2

0

3

0

3

0

Total 0 2 3 2 0 2 3 3

Epithelial hyperplasia - arytenoid

Minimal 0 1 1 0 0 0 0 2

Total 0 1 1 0 0 0 0 2

Inflammation

Minimal

Slight

0

0

0

0

1

0

1

0

0

0

0

0

0

0

0

1

Total 0 0 1 1 0 0 0 1

Number of tissues examined 3 3 3 3 3 3 3 3
Huntingdon Life Sciences

WAG0013

33

Trachea

Minimal or slight epithelial degeneration/regeneration was seen in male and female animals 

exposed to 19098 ppm and one female exposed to 8901 ppm. Minimal or slight 

inflammatory cell infiltrate in the lamina propria was seen at a higher incidence and/or degree 

in animals exposed to 19098 ppm, than in control animals.

Summary of treatment related findings in the trachea for animals killed after 2 weeks of treatment

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098

Epithelial 

degeneration/regeneration

Minimal

Slight

0

0

0

0

0

0

1

2

0

0

0

0

1

0

2

0

Total 0 0 0 3 0 0 1 2

Inflammatory cell infiltrate –

lamina propria

Minimal

Slight

1

0

1

0

0

0

1

1

0

0

0

0

1

0

0

1

Total 1 1 0 2 0 0 1 1

Number of tissues examined 3 3 3 3 3 3 3 3
Huntingdon Life Sciences

WAG0013

34

4. Discussion

The test article, 2-bromo-3,3,3-trifluoropropene, was administered to 3 groups of 6 rats by

inhalation administration, using a whole-body exposure system, for 6 hours a day, 5 days a 

week, for 2 week at achieved aerosol concentrations of 4721, 8901 and 19098 ppm. A 

control group of rats received air only at the same airflow as the high dose group.

Histopathology findings related to treatment in the nasal turbinates, were generally seen in 

the tissues of the dorsal half of the nasal cavity, with the severity of findings increasing in a 

rostral to caudal direction and the olfactory epithelium being the most extensively affected.

In the region of the olfactory epithelium, fibrosis with or without associated cartilage 

formation, atrophy of the Bowman’s glands and atrophy of olfactory nerve fibres were seen

in the dorsal lamina propria and erosion of the epithelium were seen in animals of all groups

exposed to 2-bromo-3,3,3-trifluoropropene. Necrosis of the olfactory epithelium was seen in

animals exposed to 8901 or 19098 ppm. The fibrosis with or without associated cartilage

formation is considered a protective response to irritation by the test substance, which would 

have caused the necrosis and erosion, with the atrophy of the Bowman’s glands and olfactory

nerves being either a direct effect of the irritation or a consequence of the epithelial 

degeneration.

The fusion of dorsal turbinates with associated epithelial diverticula formation seen in

animals of all test article groups, was considered a result of damage to the epithelium 

overlying the turbinates and dorsal aspect of the nasal cavity with subsequent fusion of the 

turbinates and formation of epithelial diverticula.

In the various epithelia of the nasal cavity (olfactory, respiratory and transitional) of animals 

of all test article groups similar findings were seen. These included metaplasia and

combinations of disorganisation, atrophy, degeneration and regeneration. The metaplasia

was an adaptive change to produce a type of epithelium which was more resistant to the 

irritation. The combinations of disorganisation, atrophy and degeneration were due to the 

irritation of the epithelium and the regeneration was part of the tissue repair process.

In the nasolacrimal ducts, inflammation was seen in animals of all test article groups. 

Although epithelial degeneration was seen in only one female exposed to 19098 ppm, both

the inflammation and epithelial degeneration are considered due to the irritancy of the test

substance.

In the airway, debris and inflammatory cells were seen in of all test article groups and are 

considered a consequence of the other changes in the nasal cavity.

In general, the changes seen in the nasal turbinates of animals exposed to 8901 or 19098 ppm 

were of a similar severity and incidence and of a higher severity than those seen for animals 

exposed to 4721 ppm. In consideration of the epithelial changes the olfactory epithelium was 

the most extensively affected at all three exposure levels.
Huntingdon Life Sciences

WAG0013

35

In the larynx, epithelial metaplasia, hyperplasia and keratinisation were seen in animals of all

test article groups. As with the nasal epithelia, these changes were an adaptive response to 

produce an epithelium more resistant to irritation. Inflammation was seen in one male and 

one female exposed to 19098 ppm and one male exposed to 8901 ppm and was considered a 

consequence of irritancy. In general, the findings in the larynx were of a similar incidence 

and degree in all groups exposed to 2-bromo-3,3,3-trifluoropropene.

In the trachea, epithelial degeneration/regeneration was seen in male and female animals 

exposed to 19098 ppm and one female exposed to 8901 ppm. As in the nasal cavity, the

degeneration was due to the irritation of the epithelium and the regeneration was part of the 

tissue repair process. Inflammatory cell infiltrate in the lamina propria was seen at a higher 

incidence and/or degree in animals exposed to 19098 ppm, compared with control animals, 

which again was considered a consequence of irritancy.

Overall, the most marked changes were in the nasal cavity, with minimal to slight changes in 

the larynx and trachea. For the nasal cavity, in general the changes seen in animals exposed 

to 8901 or 19098 ppm were of a similar severity and incidence. As rodents are obligate nosebreathers, this distribution of findings is considered accounted for by the nasal cavity being 

the first part of the respiratory tract to have contact with the test article. In addition, the nasal 

epithelium, and especially the olfactory epithelium which is anatomically confined to the 

dorsal half of the nasal cavity, is a site of metabolic enzymes and so metabolism of the test 

substance may have contributed to the severity of changes seen.

At necropsy, a reduction in the size of the thymus and depressions or roughening of the 

stomach were noted in animals exposed to 19098 ppm. These tissues were not required to be 

examined microscopically.

Treatment-related bodyweight losses or reductions in bodyweight gain were seen at all test 

article groups. These reductions were exposure level related and are considered to be 

treatment-related and correlate to some extent with reduced food consumption that was also 

seen.

Clinical signs of shallow breathing, piloerection, salivation and chin rubbing evident during 

and after exposure are indicative of exposure to an irritant material. Whereas, the clinical 

signs of underactivity, unsteady gait and partially closed eyelids, together with the reduced

sensory and motor activity responses seen at 8901 or 19098 ppm represent potential CNS 

effects. However, all of these signs were transient in nature.
Huntingdon Life Sciences

WAG0013

36

5. Conclusion

Histopathological changes related to treatment were seen in the nasal turbinates, larynx and 

trachea. Although the majority of these changes were limited to the 8901 or 19098 ppm

exposure groups, some changes in the nasal turbinates were evident at 4721 ppm. Therefore, 

a no observed adverse effect level (NOAEL) could not be established.
Huntingdon Life Sciences

WAG0013

37

6. References

2-Bromo-3,3,3-trifluoropropene, E-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. (Laboratory Project Number 02-5449)
38

Huntingdon Life Sciences

WAG0013

FIGURE 1

Motor activity - group mean scores (beam breaks) for males on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

 1M 2M 3M 4M

0

20

40

60

80

100

120

140

160

6 12 18 24 30 36 42 48 54 60

Number of beam breaks

Time (minutes)

High beams

0

50

100

150

200

250

300

350

6 12 18 24 30 36 42 48 54 60

Number of beam breaks

Time (minutes)

Low beams

0

250

500

750

1 2 3 4

Beam breaks

Group

Total scores

0

500

1000

1500

1 2 3 4

Beam breaks

Group

Total scores
39

Huntingdon Life Sciences

WAG0013

FIGURE1- continued

Motor activity - group mean scores (beam breaks) for females on Day 10 of treatment

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm):0 4721 8901 19098

 1F 2F 3F 4F

0

20

40

60

80

100

120

140

160

180

200

6 12 18 24 30 36 42 48 54 60

Number of beam breaks

Time (minutes)

High beams

0

50

100

150

200

250

300

350

400

6 12 18 24 30 36 42 48 54 60

Number of beam breaks

Time (minutes)

Low beams

0

250

500

750

1000

1 2 3 4

Beam breaks

Group

Total scores

0

500

1000

1500

2000

2500

1 2 3 4

Beam breaks

Group

Total scores
40

Huntingdon Life Sciences

WAG0013

FIGURE 2

Bodyweight - individual values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098
41

Huntingdon Life Sciences

WAG0013

TABLE 1

Sensory reactivity observations and grip strength - summary of findings on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group/sex: 1M 2M 3M 4M 1F 2F 3F 4F

Number of animals: 3 3 3 3 3 3 3 3

Parameter Grade

Approach response (1-3) 1 0 0 1 3 0 0 1 2

2 3 3 2 0 3 3 2 1

Touch response (1-3) 1 0 0 2 1 0 0 2 2

2 3 3 1 2 3 3 1 1

Auditory startle reflex (1-4) 1 0 0 0 2 0 0 0 0

2 1 0 1 0 0 0 0 1

3 2 3 2 1 3 3 3 2

Tail pinch response (1-4) 2 0 0 0 0 0 0 0 1

3 3 3 3 3 2 3 3 2

4 0 0 0 0 1 0 0 0

Forelimb grip strength (kg) Mean 0.94 1.14 1.06 0.95 0.92 0.94 0.81 1.04

SD 0.09 0.10 0.25 0.22 0.04 0.10 0.08 0.19

Hindlimb grip strength (kg) Mean 0.58 0.52 0.53 0.49 0.49 0.39 0.47 0.45

SD 0.01 0.05 0.03 0.06 0.11 0.07 0.03 0.11
42

Huntingdon Life Sciences

WAG0013

TABLE 2

Motor activity - group mean scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Number

of animals

Beam 

level

Time (minutes)

6 12 18 24 30 36 42 48 54 60 Total

1M 3 High 138.7 97.7 81.7 48.3 50.7 41.3 10.3 34.3 38.3 2.7 544.0

SD 18.3 12.7 60.9 47.9 38.0 43.7 13.8 32.0 65.5 2.5 297.8

2M 3 High 119.3 76.0 49.0 43.3 22.0 21.7 22.3 11.7 23.3 20.7 409.3

SD 8.4 19.1 13.1 39.2 7.0 14.8 10.1 3.2 2.1 11.0 92.4

3M 3 High 83.0 47.3 33.3 20.0 13.7 2.0 11.0 10.0 7.0 10.3 237.7

SD 11.3 28.9 30.5 19.5 19.5 2.0 9.6 11.8 6.2 17.9 108.9

4M 3 High 70.0 11.3 6.0 0.3 0.0 0.0 0.3 0.0 0.0 2.7 90.7

SD 20.7 8.1 8.7 0.6 0.0 0.0 0.6 0.0 0.0 4.6 28.7

1M 3 Low 262.3 184.3 147.7 112.3 145.0 54.3 21.0 36.3 46.0 14.7 1024.0

SD 51.0 72.6 73.5 9.1 54.1 40.4 20.5 11.7 57.7 13.3 105.3

2M 3 Low 274.7 140.3 124.3 83.3 95.0 50.3 57.0 37.7 84.3 36.7 983.7

SD 96.7 49.3 25.7 24.1 11.4 15.3 27.9 12.7 18.2 12.1 197.8

3M 3 Low 197.7 97.7 80.3 42.7 21.0 9.0 32.3 15.0 22.0 16.0 533.7

SD 52.6 50.9 62.2 25.6 19.9 4.4 31.5 13.7 26.2 27.7 95.1

4M 3 Low 179.3 63.3 20.0 1.3 0.0 0.0 1.7 4.3 2.0 23.0 295.0

SD 19.3 66.5 26.2 2.3 0.0 0.0 1.5 7.5 3.5 39.8 154.2
43

Huntingdon Life Sciences

WAG0013

TABLE 2 - continued

Motor activity - group mean scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Number

of animals

Beam 

level

Time (minutes)

6 12 18 24 30 36 42 48 54 60 Total

1F 3 High 174.7 110.7 108.0 90.3 52.3 73.0 27.7 41.0 22.0 36.3 736.0

SD 29.0 35.7 56.9 23.5 7.8 18.3 23.7 35.7 22.5 24.8 137.4

2F 3 High 101.3 73.7 44.3 22.3 24.3 16.0 6.0 2.7 24.7 27.7 343.0

SD 12.4 39.1 2.5 16.8 15.9 14.7 9.5 4.6 21.6 27.5 91.0

3F 3 High 138.3 53.3 15.7 0.0 0.0 0.0 0.0 0.0 8.7 4.3 220.3

SD 41.2 47.4 4.0 0.0 0.0 0.0 0.0 0.0 7.6 7.5 92.8

4F 3 High 121.7 26.3 13.7 0.0 3.7 9.7 0.0 0.0 0.0 0.0 175.0

SD 61.9 39.7 11.1 0.0 6.4 16.7 0.0 0.0 0.0 0.0 131.2

1F 3 Low 309.7 254.3 203.0 166.7 155.0 179.7 108.0 100.3 103.3 90.7 1670.7

SD 93.3 101.1 103.9 16.7 31.5 19.9 31.2 95.5 101.5 30.7 506.6

2F 3 Low 239.0 177.3 113.0 91.3 107.3 69.0 38.3 12.7 65.7 50.7 964.3

SD 41.1 34.7 12.5 32.7 43.3 79.6 42.1 11.7 59.2 44.0 56.6

3F 3 Low 231.3 132.0 50.0 0.7 0.0 0.3 0.0 3.0 9.0 25.3 451.7

SD 54.4 36.9 50.7 1.2 0.0 0.6 0.0 5.2 11.5 43.0 94.4

4F 3 Low 219.3 76.3 59.7 4.0 19.7 53.7 3.3 0.0 0.0 0.0 436.0

SD 88.2 114.1 60.0 6.9 34.1 93.0 5.8 0.0 0.0 0.0 374.5
44

Huntingdon Life Sciences

WAG0013

TABLE 3 Request ID: 257064

Bodyweight - group mean values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Day Day Day Day Day Day Day Day Day Day Day Day

/Sex -5 -4 -3 -2 -1 1 2 3 4 5 6 7

1M Mean 443 444 448 450 451 454 455 457 460 461 466 466

SD 20.2 15.7 16.2 13.6 15.9 17.3 20.0 17.0 19.4 15.4 20.7 18.9

N 3 3 3 3 3 3 3 3 3 3 3 3

2M Mean 391 398 404 408 418 424 406 412 420 414 414 415

SD 20.8 21.9 24.9 24.8 25.8 27.5 20.9 26.0 29.3 30.9 29.2 28.1

N 3 3 3 3 3 3 3 3 3 3 3 3

3M Mean 444 448 454 456 461 469 452 455 466 457 448 448

SD 27.3 27.2 26.8 27.2 29.6 31.3 30.9 28.7 31.7 28.7 29.1 29.6

N 3 3 3 3 3 3 3 3 3 3 3 3

4M Mean 450 450 454 453 456 453 438 436 452 438 426 417

SD 18.9 17.2 16.3 18.6 18.0 22.9 13.7 19.6 18.9 18.5 28.2 34.9

N 3 3 3 3 3 3 3 3 3 3 3 3
45

Huntingdon Life Sciences

WAG0013

TABLE 3 - continued Request ID: 257064

Bodyweight - group mean values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Day Day Day Day Day Day Day Day Change As % of

/Sex 8 9 10 11 12 13 14 15 1-15 Control

1M Mean 467 467 472 476 475 477 477 480 27 -

SD 21.1 24.8 20.8 22.5 23.1 22.6 24.7 23.1 5.8

N 3 3 3 3 3 3 3 3 3

2M Mean 425 422 438 443 439 437 429 430 6 23

SD 27.4 28.6 27.1 30.6 28.9 29.1 26.8 26.4 6.2

N 3 3 3 3 3 3 3 3 3

3M Mean 448 449 471 483 476 473 463 470 0 2

SD 27.7 23.6 30.9 29.5 33.9 26.7 24.7 29.3 5.2

N 3 3 3 3 3 3 3 3 3

4M Mean 406 400 417 434 427 419 410 410 -43 -

SD 38.4 44.9 53.6 41.6 38.3 34.3 35.2 36.3 13.5

N 3 3 3 3 3 3 3 3 3
46

Huntingdon Life Sciences

WAG0013

TABLE 3 - continued Request ID: 257064

Bodyweight - group mean values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Day Day Day Day Day Day Day Day Day Day Day Day

/Sex -5 -4 -3 -2 -1 1 2 3 4 5 6 7

1F Mean 303 298 300 292 302 304 309 298 312 315 319 304

SD 10.1 10.0 12.1 11.5 12.6 10.4 6.7 6.4 8.1 8.0 6.2 4.4

N 3 3 3 3 3 3 3 3 3 3 3 3

2F Mean 240 243 243 247 246 250 242 245 249 244 250 247

SD 9.5 6.0 3.0 7.8 9.9 3.3 7.7 10.5 12.8 13.3 10.2 11.5

N 3 3 3 3 3 3 3 3 3 3 3 3

3F Mean 258 259 257 264 265 267 261 267 271 269 267 271

SD 7.3 11.2 12.1 10.2 6.3 15.8 9.3 11.5 9.4 11.4 7.7 10.2

N 3 3 3 3 3 3 3 3 3 3 3 3

4F Mean 263 265 271 269 266 269 260 263 276 265 261 266

SD 10.2 14.0 12.4 9.2 9.4 10.0 2.4 6.6 10.3 7.3 1.9 4.4

N 3 3 3 3 3 3 3 3 3 3 3 3
47

Huntingdon Life Sciences

WAG0013

TABLE 3 - continued Request ID: 257064

Bodyweight - group mean values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Day Day Day Day Day Day Day Day Change As % of

/Sex 8 9 10 11 12 13 14 15 1-15 Control

1F Mean 320 318 317 306 315 323 315 314 10 -

SD 8.3 7.9 9.9 5.0 12.4 7.2 9.9 12.2 13.1

N 3 3 3 3 3 3 3 3 3

2F Mean 251 253 265 269 263 265 261 263 13 135

SD 12.0 10.7 16.3 12.3 6.3 10.9 13.1 11.6 8.7

N 3 3 3 3 3 3 3 3 3

3F Mean 272 274 288 292 281 287 280 284 17 168

SD 14.8 14.4 15.6 14.0 10.9 12.2 16.2 14.9 6.9

N 3 3 3 3 3 3 3 3 3

4F Mean 255 258 276 282 277 261 260 265 -3 -

SD 12.2 12.5 5.3 5.1 5.5 6.1 15.5 6.5 16.5

N 3 3 3 3 3 3 3 3 3
48

Huntingdon Life Sciences

WAG0013

TABLE 4 Request ID: 257053

Organ weights - group mean unadjusted and adjusted values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Terminal 

Bodyweight

Brain Heart Kidneys Liver Lungs + 

Bronchi

Spleen

Unadjusted Means

1M Mean 483 2.04 1.690 3.15 17.30 1.640 0.790

SD 24 0.05 0.155 0.53 1.54 0.166 0.017

N 3 3 3 3 3 3 3

2M Mean 432 2.01 1.456 3.44 15.72 1.634 0.663

SD 25 0.02 0.264 0.25 2.20 0.099 0.090

N 3 3 3 3 3 3 3

3M Mean 469 2.07 1.635 3.21 19.75 1.658 0.891

SD 28 0.10 0.262 0.35 2.20 0.282 0.164

N 3 3 3 3 3 3 3

4M Mean 412 2.09 1.569 2.97 16.15 1.730 0.636

SD 36 0.11 0.199 0.54 1.30 0.258 0.055

N 3 3 3 3 3 3 3

Adjusted Means

1M Mean 1.96 1.714 2.74 15.93 1.517 0.728

2M Mean 2.05 1.444 3.65 16.41 1.696 0.694

3M Mean 2.02 1.650 2.96 18.92 1.584 0.854

4M Mean 2.18 1.542 3.42 17.65 1.864 0.704
49

Huntingdon Life Sciences

WAG0013

TABLE 4 - continued Request ID: 257053

Organ weights - group mean unadjusted and adjusted values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Terminal 

Bodyweight

Brain Heart Kidneys Liver Lungs + 

Bronchi

Spleen

Unadjusted Means

1F Mean 314 1.97 1.325 1.91 10.12 1.368 0.563

SD 12 0.02 0.100 0.07 0.83 0.097 0.046

N 3 3 3 3 3 3 3

2F Mean 264 1.84 0.927 1.89 9.30 1.190 0.510

SD 11 0.14 0.111 0.18 1.23 0.100 0.084

N 3 3 3 3 3 3 3

3F Mean 283 1.88 1.049 1.91 11.51 1.298 0.502

SD 15 0.13 0.112 0.13 0.88 0.202 0.105

N 3 3 3 3 3 3 3

4F Mean 265 1.87 1.029 1.72 10.08 1.328 0.484

SD 7 0.03 0.036 0.05 0.25 0.117 0.031

N 3 3 3 3 3 3 3

Adjusted Means

1F Mean 1.79 1.250 1.91 8.67 1.349 0.712

2F Mean 1.93 0.967 1.90 10.07 1.200 0.430

3F Mean 1.87 1.046 1.91 11.44 1.297 0.510

4F Mean 1.96 1.068 1.72 10.82 1.337 0.408
50

Huntingdon Life Sciences

WAG0013

TABLE 5 Request ID: 264436

Macropathology - group distribution of findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group/Sex: 1M 2M 3M 4M 1F 2F 3F 4F

Tissue and Finding Number Examined: 3 3 3 3 3 3 3 3

Kidneys

Pale area(s) 0 0 0 1 0 0 0 0

Pelvic dilatation 0 0 0 1 0 0 0 0

Ln mandibular

Enlarged 0 0 0 0 0 1 0 0

Lungs + bronchi

Dark area(s) 0 0 1 0 0 0 0 0

Pale area(s) 0 1 0 1 0 1 0 0

Skin

Hair loss 2 0 2 0 0 0 0 0

Scab(s) 0 0 1 0 0 0 0 0

Spleen

Capsule thickened 0 2 0 0 0 0 0 0

Stomach

Forestomach depression(s) 0 0 0 2 0 0 0 0

Forestomach roughened 0 0 0 1 0 0 0 2
51

Huntingdon Life Sciences

WAG0013

TABLE 5 - continued Request ID: 264436

Macropathology - group distribution of findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group/Sex: 1M 2M 3M 4M 1F 2F 3F 4F

Tissue and Finding Number Examined: 3 3 3 3 3 3 3 3

Thymus

Small 0 0 0 3 0 0 0 3

Ln bronchial

Enlarged 0 0 0 0 0 1 0 0
52

Huntingdon Life Sciences

WAG0013

TABLE 6 Request ID: 264437

Histopathology - group distribution of findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group/Sex: 1M 2M 3M 4M 1F 2F 3F 4F

Tissue and Finding Number: 3 3 3 3 3 3 3 3

Trachea Number Examined: 3 3 3 3 3 3 3 3

Epithelial Degeneration/Regeneration 0 0 0 3 0 0 1 2

Inflammatory Cell Infiltrate - Lamina Propria 1 1 0 2 0 0 1 1

Tracheal Bifurc. Number Examined: 3 3 3 3 3 3 3 3

Inflammatory Cell Infiltrate in Lamina Propria 2 1 0 2 0 0 1 1

Lungs + Bronchi Number Examined: 3 0 0 3 3 0 0 3

Alveolar Eosinophilic Crystals with Associated Inflammatory 

Cell Infiltrate

0 0 0 0 2 0 0 0

Alveolar Macrophages 1 0 0 1 1 0 0 3

Alveolitis 1 0 0 1 0 0 0 0

Arterial Mural Mineralisation 2 0 0 2 2 0 0 1

Perivascular Inflammatory Cell Infiltrate 2 0 0 0 0 0 0 0

Larynx Number Examined: 3 3 3 3 3 3 3 3

Epithelial Hyperplasia - Arytenoid 0 1 1 0 0 0 0 2

Epithelial Hyperplasia - Ventral 0 0 0 1 0 0 0 0

Epithelial Keratinisation - Ventrolateral 0 2 2 3 0 2 2 3

Inflammation 0 0 1 1 0 0 0 1

Squamous Metaplasia - Arytenoid 0 2 3 2 0 2 3 3

Squamous Metaplasia - Ventral 0 0 1 2 0 0 3 3

Submucosal Foreign Body Granulomata - Ventral 0 1 1 0 0 0 0 0
53

Huntingdon Life Sciences

WAG0013

TABLE 6 - continued Request ID: 264437

Histopathology - group distribution of findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group/Sex: 1M 2M 3M 4M 1F 2F 3F 4F

Tissue and Finding Number: 3 3 3 3 3 3 3 3

Nasal turbinates Number Examined: 3 3 3 3 3 3 3 3

Debris and Inflammatory Cells in Airway 0 2 1 1 0 0 2 1

Fusion of Dorsal Turbinate With Associated Epithelial 

Diverticula Formation

0 0 3 3 0 1 3 3

Nasolacrimal Ducts - Epithelial Degeneration 0 0 0 0 0 0 0 1

Nasolacrimal Ducts - Inflammation 0 1 1 2 0 1 0 2

Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal 

Lamina Propria

0 3 3 3 0 3 3 3

Olfactory Epithelium - Degeneration/Atrophy/Disorganisation 0 3 3 3 0 2 3 3

Olfactory Epithelium - Erosion 0 1 1 2 0 1 1 1

Olfactory Epithelium - Fibrosis With or Without Associated 

Cartilage Formation in Dorsal Lamina Propria

0 2 3 3 0 3 3 3

Olfactory Epithelium - Necrosis 0 0 0 2 0 0 1 0

Olfactory Epithelium - Respiratory Metaplasia With or Without 

Associated Inflammatory Cell Infiltrate

0 2 3 3 0 3 3 2

Olfactory Epithelium - Squamous Metaplasia 0 3 2 1 0 3 3 2

Olfactory Nerve Fibres - Atrophy 0 3 3 3 0 3 3 3

Respiratory Epithelium - Disorganisation/Degeneration/ 

Regeneration

0 1 3 3 0 1 2 3

Respiratory Epithelium - Goblet Cell Hyperplasia 0 1 1 0 0 1 1 0

Respiratory Epithelium - Squamous Metaplasia 0 0 0 3 0 0 0 1

Submucosal Foreign Body Granulomata - Dorsal 0 0 1 1 0 0 0 0

Transitional Epithelium - Disorganisation/Regeneration 0 2 2 3 0 2 2 3
54

Huntingdon Life Sciences

WAG0013

APPENDIX 1 Request ID: 258669

Clinical signs - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Death Day of

/Sex Number Code Death Category Observation Day(s)

1M 1 T 15 Coat Hair loss, Forelimbs -4, 3, 10, 14-15

3 T 15 Coat Hair loss, Forelimbs 14-15

Only animals with observations are presented
55

Huntingdon Life Sciences

WAG0013

APPENDIX 1 - continued Request ID: 258669

Clinical signs - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Death Day of

/Sex Number Code Death Category Observation Day(s)

2M 5 T 15 Coat Hair loss, Forepaws -4, 3, 10

Only animals with observations are presented
56

Huntingdon Life Sciences

WAG0013

APPENDIX 1 - continued Request ID: 258669

Clinical signs - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Death Day of

/Sex Number Code Death Category Observation Day(s)

3M 7 T 15 Coat Hair loss, Head -4, 3, 10, 14-15

9 T 15 Coat Hair loss, Head 3, 10, 14-15

Only animals with observations are presented
57

Huntingdon Life Sciences

WAG0013

APPENDIX 1 - continued Request ID: 258669

Clinical signs - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Death Day of

/Sex Number Code Death Category Observation Day(s)

4M 10 T 15 Behaviour Underactive 15

11 T 15 Coat Piloerection 10

Only animals with observations are presented
58

Huntingdon Life Sciences

WAG0013

APPENDIX 1 - continued Request ID: 258669

Clinical signs - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Death Day of

/Sex Number Code Death Category Observation Day(s)

1F 14 T 15 Behaviour Irritable 10, 14

Only animals with observations are presented
59

Huntingdon Life Sciences

WAG0013

APPENDIX 1 - continued Request ID: 258669

Clinical signs - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Death Day of

/Sex Number Code Death Category Observation Day(s)

4F 22 T 15 Behaviour Underactive 14-15

Coat Piloerection 14-15

Staining Abnormal Colour, Red, Head 14-15

23 T 15 Body temperature Reduced 14

Coat Hair loss, Forelimbs -4, 3, 10, 14-15

Muscle Reaction Abnormal Gait, Elevated 14-15

Staining Abnormal Colour, Brown, Anus 14

24 T 15 Behaviour Underactive 14-15

Only animals with observations are presented
60

Huntingdon Life Sciences

WAG0013

APPENDIX 2 Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of

dosing

1M 1 Coat

 Piloerection 5 1, 4-5 1, 4-5

2 Coat

 Piloerection 1 1
61

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

1M 1 Coat

 Piloerection

2 Coat

 Piloerection
62

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

2M 4 Behaviour

 Chin Rubbing 13-14 12 12 11

 Salivation 1, 11, 13 13

Breathing

 Gasping 1

 Shallow 1 1 1, 12

 Slow 1

Coat

 Piloerection 1, 4 1, 12 1, 12 14

Eyelids (Left)

 Closed 1

 Partially Closed 11, 14 11-12, 14 12, 14 14

Eyelids (Right)

 Closed 1

 Partially Closed 11, 14 11-12, 14 12, 14 14

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Posture

 Hunched 1 1 1
63

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

2M 4 Behaviour

 Chin Rubbing

 Salivation

Breathing

 Gasping

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Closed

 Partially Closed

Eyelids (Right)

 Closed

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Posture

 Hunched
64

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

2M 5 Behaviour

 Chin Rubbing 8, 12-14 12 7-8, 11-14

 Irritable 13

 Salivation 1, 8, 11-14 8, 11-14 7, 12-14 7, 12-14

Breathing

 Gasping 1

 Shallow 1, 12 1, 12 1, 12

 Slow 1

Coat

 Piloerection 1 1 1 14

Eyelids (Left)

 Closed 1

 Partially Closed 14 12-14 12-14 14

Eyelids (Right)

 Closed 1

 Partially Closed 14 12-14 12-14 14

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Posture

 Hunched 1 1 1

Staining

 Post Salivation Staining 8 8, 11, 14
65

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

2M 5 Behaviour

 Chin Rubbing

 Irritable

 Salivation

Breathing

 Gasping

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Closed

 Partially Closed

Eyelids (Right)

 Closed

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Posture

 Hunched

Staining

 Post Salivation Staining 7, 11 7, 11
66

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

2M 6 Behaviour

 Chin Rubbing 8, 11, 13-14

 Salivation 1, 11, 13

Breathing

 Gasping 1

 Shallow 1 1, 12 1, 12

 Slow 1

Coat

 Piloerection 1, 14 1, 12, 14 1, 12, 14

Eyelids (Left)

 Closed 1

 Partially Closed 14 11-14 12, 14

Eyelids (Right)

 Closed 1

 Partially Closed 14 11-14 12, 14

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Posture

 Hunched 1 1 1

Staining

 Post Salivation Staining 11
67

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

2M 6 Behaviour

 Chin Rubbing

 Salivation

Breathing

 Gasping

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Closed

 Partially Closed

Eyelids (Right)

 Closed

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Posture

 Hunched

Staining

 Post Salivation Staining
68

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3M 7 Behaviour

 Chin Rubbing 5-8, 11 5 8

 Irritable 6

 Underactive 8, 12, 14 8, 12-14 12-14 11

 Unresponsive 6-7 6

 Salivation 1, 5-6, 11-14 5, 13-14 5, 13-14 13

Breathing

 Gasping 1

 Irregular 1

 Shallow 1, 4-5, 7, 11 1, 4-7, 11 1, 5-7, 11

 Slow 1 1

Coat

 Piloerection 8, 11 8, 11-12 8, 11-12 8

Eyelids (Left)

 Partially Closed 1, 6-8, 11-14 6-8, 11-14 6-8, 11-14

Eyelids (Right)

 Partially Closed 1, 6-8, 11-14 6-8, 11-14 6-8, 11-14

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Muscle Reaction

 Unsteady 1, 8 8 4
69

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3M 7 Behaviour

 Chin Rubbing

 Irritable

 Underactive

 Unresponsive

 Salivation

Breathing

 Gasping

 Irregular

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Unsteady
70

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3M 7 Staining

 Post Salivation Staining 5-8, 11
71

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3M 7 Staining

 Post Salivation Staining
72

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3M 8 Behaviour

 Chin Rubbing 5-7, 11 5-6 6 6

 Underactive 8, 12-14 8, 12-14 12-14

 Unresponsive 6-7 6

 Salivation 1 5 5

Breathing

 Gasping 1

 Irregular 1

 Shallow 1, 4-5, 7, 11 1, 4-7, 11 1, 5-7, 11 7

 Slow 1

Coat

 Piloerection 8, 11 8, 11-12 7-8, 11-12 7

Eyelids (Left)

 Partially Closed 1, 7-8, 11-14 1, 6-8, 11, 1, 6-8, 11, 7

13-14 13-14

Eyelids (Right)

 Partially Closed 1, 7-8, 11-14 1, 6-8, 11, 1, 6-8, 11, 7

13-14 13-14

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Muscle Reaction

 Unsteady 8
73

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3M 8 Behaviour

 Chin Rubbing

 Underactive

 Unresponsive

 Salivation

Breathing

 Gasping

 Irregular

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Unsteady
74

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3M 8 Staining

 Post Salivation Staining 5-7
75

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3M 8 Staining

 Post Salivation Staining
76

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3M 9 Behaviour

 Chin Rubbing 5-7 5 4

 Underactive 8, 12, 14 8, 12-14 12-14 11

 Unresponsive 6-7 6

 Salivation 1

Breathing

 Gasping 1

 Irregular 1

 Shallow 1, 4, 7, 11 1, 4-7, 11 1, 5-6, 11

 Slow 1 1

Coat

 Piloerection 8, 12 5, 8, 12 5, 8, 12 7

Eyelids (Left)

 Closed 1

 Partially Closed 1, 5-8, 11-14 5-8, 11-14 5-6, 8, 11-14

Eyelids (Right)

 Closed 1

 Partially Closed 1, 5-8, 11-14 5-8, 11-14 5-6, 8, 11-14

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Muscle Reaction

 Unsteady 1 8 4
77

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3M 9 Behaviour

 Chin Rubbing

 Underactive

 Unresponsive

 Salivation

Breathing

 Gasping

 Irregular

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Closed

 Partially Closed

Eyelids (Right)

 Closed

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Unsteady
78

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

4M 10 Behaviour

 Irritable 4

 Underactive 1, 5-7 5-7 4-7

 Unresponsive 1, 4 4, 7 7

 Salivation 1

Breathing

 Gasping 1

 Shallow 1, 4-5 1, 4-5, 7 1, 4-5, 7

 Slow 1 7 7

Coat

 Piloerection 1, 6-7 1, 5-7 1, 4-7 1, 4

Eyelids (Left)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 1

Eyelids (Right)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 1

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Muscle Reaction

 Unsteady 1, 6 1, 5-6 1, 6 6
79

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

4M 10 Behaviour

 Irritable

 Underactive

 Unresponsive

 Salivation

Breathing

 Gasping

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Unsteady
80

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

4M 11 Behaviour

 Underactive 1, 5-7 5-7 4-7 5

 Unresponsive 1, 7 7 7

 Salivation 1

Body Temperature

 Reduced

Breathing

 Gasping 1

 Shallow 1, 4-5, 7 1, 4-5, 7 1, 4-5, 7

 Slow 1

Coat

 Piloerection 1, 6 1, 5-6 1, 4-6 4, 6-7

Eyelids (Left)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 4-5

Eyelids (Right)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 4-5

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Muscle Reaction

 Body Tremors 7

 Unsteady 1, 6 1, 6 1, 6 6-7
81

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

4M 11 Behaviour

 Underactive

 Unresponsive

 Salivation

Body Temperature

 Reduced 8 8

Breathing

 Gasping

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Body Tremors

 Unsteady
82

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

4M 11 Posture

 Hunched 7

12 Behaviour

 Irritable 4

 Underactive 1, 5-7 5-7 4-7 5

 Unresponsive 1

 Salivation 1

Breathing

 Gasping 1

 Shallow 1, 4-5 1, 4-5 1, 4-5

 Slow 1

Coat

 Piloerection 1, 5-6 1, 5-6 1, 4-6 4, 6

Eyelids (Left)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 4-5

Eyelids (Right)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 4-5

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Muscle Reaction

 Unsteady 1, 6 1, 5-6 1, 6 6
83

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

4M 11 Posture

 Hunched

12 Behaviour

 Irritable

 Underactive

 Unresponsive

 Salivation

Breathing

 Gasping

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Unsteady
84

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

1F 13 Coat

 Piloerection 7 1, 4-5, 7 1, 4-7

Eyelids (Left)

 Partially Closed 7 7

Eyelids (Right)

 Partially Closed 7 7

14 Coat

 Piloerection 1, 6

15 Coat

 Piloerection 4 4, 6
85

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

1F 13 Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

14 Coat

 Piloerection

15 Coat

 Piloerection
86

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

2F 16 Behaviour

 Chin Rubbing 8, 11-14 11-12 12 8, 13-14

 Irritable 12 12

 Underactive 7

 Unresponsive 13 13

 Salivation 1, 11-13 12 12

Body Temperature

 Reduced 7 7

Breathing

 Gasping 1

 Irregular 7

 Shallow 1, 11-12, 14 1, 7, 11-14 1, 7, 12-14 7

Coat

 Piloerection 1, 4, 7, 14 1, 7, 12-14 1, 7, 12-14 7, 14

Eyelids (Left)

 Closed 1

 Partially Closed 7, 11, 13-14 7, 11, 13-14 7, 12-14 7

Eyelids (Right)

 Closed 1

 Partially Closed 7, 11, 13-14 7, 11, 13-14 7, 12-14 7

Eye (Left)

 Dull 7

 Lacrimation 1
87

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

2F 16 Behaviour

 Chin Rubbing

 Irritable

 Underactive 7 7

 Unresponsive

 Salivation

Body Temperature

 Reduced

Breathing

 Gasping

 Irregular

 Shallow

Coat

 Piloerection

Eyelids (Left)

 Closed

 Partially Closed

Eyelids (Right)

 Closed

 Partially Closed

Eye (Left)

 Dull 7 7

 Lacrimation
88

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

2F 16 Eye (Right)

 Dull 7

 Lacrimation 1

Muscle Reaction

 Abnormal Gait

 Unsteady 7

Posture

 Hunched 1 1 1 7

Skin Colour

 Pallor 7 7 7

Staining

 Post Salivation Staining 8, 12

Comments:

 Elevated
89

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

2F 16 Eye (Right)

 Dull 7 7

 Lacrimation

Muscle Reaction

 Abnormal Gait 7 7

 Unsteady 7 7

Posture

 Hunched 7 7

Skin Colour

 Pallor 7 7

Staining

 Post Salivation Staining

Comments:

 Elevated 7 7
90

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

2F 17 Behaviour

 Chin Rubbing 8, 11-14 11 11, 14

 Unresponsive 13 13

 Salivation 1, 11, 13

Breathing

 Gasping 1

 Shallow 1, 11 1, 11-13 1, 12-13

Coat

 Piloerection 1 1, 12 1, 12, 14 14

Eyelids (Left)

 Closed 1

 Partially Closed 11, 13-14 11, 13 12-13 14

Eyelids (Right)

 Closed 1

 Partially Closed 11, 13-14 11, 13 12-13 14

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Posture

 Hunched 1 1 1

Staining

 Post Salivation Staining 14
91

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

2F 17 Behaviour

 Chin Rubbing

 Unresponsive

 Salivation

Breathing

 Gasping

 Shallow

Coat

 Piloerection

Eyelids (Left)

 Closed

 Partially Closed

Eyelids (Right)

 Closed

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Posture

 Hunched

Staining

 Post Salivation Staining
92

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

2F 18 Behaviour

 Chin Rubbing 8, 11-14 8, 11 12 14

 Irritable 13

 Unresponsive 13

 Salivation 1, 11-13 12-13 12

Breathing

 Gasping 1

 Shallow 1, 11 1, 11-12 1, 12

Coat

 Piloerection 1, 4, 14 1, 7, 12-14 1, 4, 7, 11-14 11, 14

Eyelids (Left)

 Closed 1

 Partially Closed 11, 13-14 11, 13-14 13-14

Eyelids (Right)

 Closed 1

 Partially Closed 11, 13-14 11, 13-14 13-14

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Posture

 Hunched 1 1 1
93

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

2F 18 Behaviour

 Chin Rubbing

 Irritable

 Unresponsive

 Salivation

Breathing

 Gasping

 Shallow

Coat

 Piloerection

Eyelids (Left)

 Closed

 Partially Closed

Eyelids (Right)

 Closed

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Posture

 Hunched
94

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3F 19 Behaviour

 Chin Rubbing 4-8, 11-13 5 6, 8

 Irritable 4

 Underactive 8, 12-14 8, 12-14 12-14 7, 11

 Unresponsive 6-7 6-7

 Salivation 1, 6

Breathing

 Gasping 1

 Irregular 1

 Shallow 1, 4-5, 7, 11 1, 4-7, 11 1, 5-7, 11

 Slow 1

Coat

 Piloerection 7-8, 11-14 1, 5, 7-8, 1, 5-8, 11-14 4, 7-8, 11

11-14

Eyelids (Left)

 Partially Closed 1, 4-8, 12-14 1, 5-8, 12-14 1, 5-8, 12-14 7

Eyelids (Right)

 Partially Closed 1, 4-8, 12-14 1, 5-8, 12-14 1, 5-8, 12-14 7

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1

Muscle Reaction

 Unsteady 1
95

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3F 19 Behaviour

 Chin Rubbing

 Irritable 4 4

 Underactive

 Unresponsive

 Salivation

Breathing

 Gasping

 Irregular

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Unsteady
96

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3F 20 Behaviour

 Chin Rubbing 6-8, 11 6 6, 13

 Irritable 4

 Underactive 8, 12-14 8, 12-14 12-14 7, 11-12

 Unresponsive 6 6-7 6-7

 Salivation 1

Breathing

 Gasping 1

 Irregular 1

 Shallow 1, 4-5, 7, 11 1, 4-7, 11-12 1, 5-7, 11-12

 Slow 1

Coat

 Piloerection 8 5, 8, 12-13 5, 8, 11-13

Eyelids (Left)

 Partially Closed 1, 7-8, 12-14 1, 7-8, 12-14 1, 5, 7-8, 7

12-14

Eyelids (Right)

 Partially Closed 1, 7-8, 12-14 1, 7-8, 12-14 1, 5, 7-8, 7

12-14

Eye (Left)

 Lacrimation 1 13

Eye (Right)

 Lacrimation 1 13
97

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3F 20 Behaviour

 Chin Rubbing

 Irritable

 Underactive

 Unresponsive

 Salivation

Breathing

 Gasping

 Irregular

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation
98

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3F 20 Muscle Reaction

 Unsteady 1

Staining

 Post Salivation Staining 1
99

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3F 20 Muscle Reaction

 Unsteady

Staining

 Post Salivation Staining
100

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3F 21 Behaviour

 Chin Rubbing 6-8, 11-12 6, 13

 Irritable 4

 Underactive 8, 12, 14 8, 12-14 12-14 11

 Unresponsive 6-7 6-7

 Salivation 1

Breathing

 Gasping 1

 Irregular 1

 Shallow 1, 4-5, 7, 11 1, 4-7, 11-12 1, 5-7, 11-12

 Slow 1

Coat

 Piloerection 7-8, 11-14 1, 5, 7-8, 1, 5-8, 11-14 7-8

11-14

Eyelids (Left)

 Closed 1

 Partially Closed 1, 5-8, 11-14 1, 5-8, 11-14 1, 5-8, 12-14 8

Eyelids (Right)

 Closed 1

 Partially Closed 1, 5-8, 11-14 1, 5-8, 11-14 1, 5-8, 12-14 8

Eye (Left)

 Lacrimation 1

Eye (Right)

 Lacrimation 1
101

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3F 21 Behaviour

 Chin Rubbing

 Irritable

 Underactive

 Unresponsive

 Salivation

Breathing

 Gasping

 Irregular

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Closed

 Partially Closed

Eyelids (Right)

 Closed

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation
102

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

3F 21 Muscle Reaction

 Unsteady 1
103

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

3F 21 Muscle Reaction

 Unsteady
104

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

4F 22 Behaviour

 Irritable 4

 Underactive 1, 5, 7 1, 5, 7 4-5

 Unresponsive 1, 4 1, 4

 Salivation 1 1

Breathing

 Gasping 1

 Shallow 1, 4-7 1, 4-7 1, 4-7 4

 Slow 1 1

Coat

 Piloerection 1, 4-7 1, 4-7 1, 4-7 1, 4, 6

Eyelids (Left)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7

Eyelids (Right)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7

Eye (Left)

 Lacrimation 1 1

Eye (Right)

 Lacrimation 1 1

Muscle Reaction

 Unsteady 1, 4-7 1, 5-7 1, 6 6
105

Huntingdon Life Sciences

WAG0013

APPENDIX 2- continued Request ID: 259116

Signs associated with dosing -individual observations

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm):0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

4F 22 Behaviour

 Irritable

 Underactive

 Unresponsive

 Salivation

Breathing

 Gasping

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Unsteady
106

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

4F 23 Behaviour

 Irritable 4

 Underactive 1, 5-7 1, 5-7 5-7

 Unresponsive 1, 4 1, 4

 Salivation 1 1

Breathing

 Shallow 1, 4-7 1, 4-7 1, 4-7 4

 Slow 1 1

Coat

 Piloerection 1, 4-7 1, 4-7 1, 4-7 1, 4, 6

Eyelids (Left)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 5

Eyelids (Right)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 5

Eye (Left)

 Lacrimation 1 1

Eye (Right)

 Lacrimation 1 1

Muscle Reaction

 Unsteady 1, 4, 6 1, 6 1, 4, 6 6
107

Huntingdon Life Sciences

WAG0013

APPENDIX 2- continued Request ID: 259116

Signs associated with dosing - individual observations

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm):0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

4F 23 Behaviour

 Irritable

 Underactive

 Unresponsive

 Salivation

Breathing

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Unsteady
108

Huntingdon Life Sciences

WAG0013

APPENDIX 2 - continued Request ID: 259116

Signs associated with dosing - individual observations

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign Pre-dose

1 to 2 hours 

during exposure

3 to 4 hours 

during exposure

5 to 6 hours 

during exposure

1 to 2 hours after 

completion of 

dosing

4F 24 Behaviour

 Irritable 4

 Underactive 1, 5-7 1, 5-7 5-7 5-6

 Unresponsive 4 4

 Salivation 1 1

Breathing

 Shallow 1, 4, 6 1, 4, 6 1, 4, 6 4

 Slow 1 1

Coat

 Piloerection 1, 5-7 1, 5-7 1, 4-7 1, 4, 6

Eyelids (Left)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 4-5

Eyelids (Right)

 Partially Closed 1, 4-7 1, 4-7 1, 4-7 4-5

Eye (Left)

 Lacrimation 1 1

Eye (Right)

 Lacrimation 1 1

Muscle Reaction

 Unsteady 1, 4, 6 1, 6 1, 6 6
109

Huntingdon Life Sciences

WAG0013

APPENDIX2- continued Request ID: 259116

Signs associated with dosing - individual observations

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm):0 4721 8901 19098

Days

Group 

/Sex

Animal 

Number Sign

Additional 

observation

As late as 

possible in the 

working day

4F 24 Behaviour

 Irritable

 Underactive

 Unresponsive

 Salivation

Breathing

 Shallow

 Slow

Coat

 Piloerection

Eyelids (Left)

 Partially Closed

Eyelids (Right)

 Partially Closed

Eye (Left)

 Lacrimation

Eye (Right)

 Lacrimation

Muscle Reaction

 Unsteady
110

Huntingdon Life Sciences

WAG0013

APPENDIX 3

Sensory reactivity observations and grip strength - individual findings on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group/sex: 1M 2M 3M 4M

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

Parameter (Grade range)

Approach response (1-3) 2 2 2 2 2 2 1 2 2 1 1 1

Touch response (1-3) 2 2 2 2 2 2 1 2 1 1 2 2

Auditory startle reflex (1-4) 2 3 3 3 3 3 2 3 3 3 1 1

Tail pinch response (1-4) 3 3 3 3 3 3 3 3 3 3 3 3

Forelimb grip strength (kg) Trial 1 1.01 1.14 0.88 1.36 1.22 1.07 1.52 0.90 0.98 0.65 0.95 1.18

Trial 2 0.85 0.98 0.71 1.21 1.15 0.98 1.18 0.86 0.93 0.68 0.85 1.07

Trial 3 1.04 0.89 0.94 1.18 0.91 1.17 1.34 0.91 0.90 0.82 1.12 1.21

Mean 0.97 1.00 0.84 1.25 1.09 1.07 1.35 0.89 0.94 0.72 0.97 1.15

Hindlimb grip strength (kg) Trial 1 0.51 0.61 0.62 0.51 0.58 0.50 0.56 0.49 0.60 0.39 0.41 0.58

Trial 2 0.58 0.59 0.58 0.44 0.55 0.60 0.57 0.50 0.51 0.57 0.45 0.62

Trial 3 0.65 0.53 0.56 0.48 0.59 0.42 0.56 0.50 0.52 0.51 0.46 0.45

Mean 0.58 0.58 0.59 0.48 0.57 0.51 0.56 0.50 0.54 0.49 0.44 0.55

Additional observations - - - - - - - - - - - -
111

Huntingdon Life Sciences

WAG0013

APPENDIX 3 - continued

Sensory reactivity observations and grip strength - individual findings on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group/sex: 1F 2F 3F 4F

Animal number: 13 14 15 16 17 18 19 20 21 22 23 24

Parameter (Grade range)

Approach response (1-3) 2 2 2 2 2 2 2 2 1 1 1 2

Touch response (1-3) 2 2 2 2 2 2 2 1 1 1 1 2

Auditory startle reflex (1-4) 3 3 3 3 3 3 3 3 3 3 3 2

Tail pinch response (1-4) 4 3 3 3 3 3 3 3 3 2 3 3

Forelimb grip strength (kg) Trial 1 0.94 0.88 0.89 1.08 0.80 1.04 0.97 0.82 0.72 0.94 1.10 1.21

Trial 2 0.99 0.96 0.96 1.03 0.76 0.95 0.76 0.72 0.78 0.86 1.12 1.04

Trial 3 0.94 0.84 0.85 0.84 0.89 1.03 0.94 0.89 0.73 0.70 1.07 1.34

Mean 0.96 0.89 0.90 0.98 0.82 1.01 0.89 0.81 0.74 0.83 1.10 1.20

Hindlimb grip strength (kg) Trial 1 0.62 0.45 0.37 0.38 0.31 0.47 0.44 0.48 0.44 0.47 0.39 0.50

Trial 2 0.58 0.47 0.44 0.40 0.32 0.45 0.43 0.42 0.62 0.42 0.32 0.54

Trial 3 0.62 0.48 0.36 0.40 0.32 0.42 0.46 0.50 0.42 0.44 0.34 0.66

Mean 0.61 0.47 0.39 0.39 0.32 0.45 0.44 0.47 0.49 0.44 0.35 0.57

Additional observations - - - - - - - - - - - -
112

Huntingdon Life Sciences

WAG0013

APPENDIX 4

Motor activity - individual scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal Beam 

level

Time (minutes)

number 6 12 18 24 30 36 42 48 54 60 Total

1M 1 High 145 83 37 28 23 37 26 20 0 5 404

2 118 105 57 14 35 0 0 12 1 0 342

3 153 105 151 103 94 87 5 71 114 3 886

1 Low 261 160 97 102 197 89 41 23 21 18 1009

2 314 266 232 119 149 10 0 41 5 0 1136

3 212 127 114 116 89 64 22 45 112 26 927
113

Huntingdon Life Sciences

WAG0013

APPENDIX 4 - continued

Motor activity - individual scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal Beam 

level

Time (minutes)

number 6 12 18 24 30 36 42 48 54 60 Total

2M 4 High 129 88 64 87 15 38 34 8 21 32 516

5 115 54 43 32 22 18 16 13 25 20 358

6 114 86 40 11 29 9 17 14 24 10 354

4 Low 192 97 111 106 82 68 25 23 64 24 792

5 251 130 108 86 103 42 70 44 90 48 972

6 381 194 154 58 100 41 76 46 99 38 1187
114

Huntingdon Life Sciences

WAG0013

APPENDIX 4 - continued

Motor activity - individual scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal Beam 

level

Time (minutes)

number 6 12 18 24 30 36 42 48 54 60 Total

3M 7 High 89 62 33 39 36 4 15 23 9 31 341

8 90 66 64 21 5 2 0 0 0 0 248

9 70 14 3 0 0 0 18 7 12 0 124

7 Low 230 62 49 31 44 6 34 27 15 48 546

8 226 156 152 72 9 7 0 0 0 0 622

9 137 75 40 25 10 14 63 18 51 0 433
115

Huntingdon Life Sciences

WAG0013

APPENDIX 4 - continued

Motor activity - individual scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal Beam 

level

Time (minutes)

number 6 12 18 24 30 36 42 48 54 60 Total

4M 10 High 48 10 1 0 0 0 1 0 0 0 60

11 89 4 1 1 0 0 0 0 0 0 95

12 73 20 16 0 0 0 0 0 0 8 117

10 Low 157 29 2 0 0 0 2 0 0 0 190

11 190 21 8 4 0 0 0 0 0 0 223

12 191 140 50 0 0 0 3 13 6 69 472
116

Huntingdon Life Sciences

WAG0013

APPENDIX 4 - continued

Motor activity - individual scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal Beam 

level

Time (minutes)

number 6 12 18 24 30 36 42 48 54 60 Total

1F 13 High 145 98 67 67 50 77 55 0 0 23 582

14 203 151 84 90 61 53 14 58 45 21 780

15 176 83 173 114 46 89 14 65 21 65 846

13 Low 224 160 92 148 124 173 141 5 0 70 1137

14 409 361 298 180 154 164 104 196 203 76 2145

15 296 242 219 172 187 202 79 100 107 126 1730
117

Huntingdon Life Sciences

WAG0013

APPENDIX 4 - continued

Motor activity - individual scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal Beam 

level

Time (minutes)

number 6 12 18 24 30 36 42 48 54 60 Total

2F 16 High 108 116 47 26 6 0 0 0 40 55 398

17 109 66 42 37 33 19 17 8 34 28 393

18 87 39 44 4 34 29 1 0 0 0 238

16 Low 286 173 127 61 73 2 14 15 82 66 899

17 210 145 103 87 93 48 87 23 115 85 996

18 221 214 109 126 156 157 14 0 0 1 998
118

Huntingdon Life Sciences

WAG0013

APPENDIX 4 - continued

Motor activity - individual scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal Beam 

level

Time (minutes)

number 6 12 18 24 30 36 42 48 54 60 Total

3F 19 High 184 108 18 0 0 0 0 0 14 0 324

20 127 29 11 0 0 0 0 0 12 13 192

21 104 23 18 0 0 0 0 0 0 0 145

19 Low 211 147 33 2 0 1 0 9 5 0 408

20 190 90 10 0 0 0 0 0 22 75 387

21 293 159 107 0 0 0 0 0 0 1 560
119

Huntingdon Life Sciences

WAG0013

APPENDIX 4 - continued

Motor activity - individual scores (beam breaks) on Day 10 of treatment

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal Beam 

level

Time (minutes)

number 6 12 18 24 30 36 42 48 54 60 Total

4F 22 High 187 72 24 0 11 29 0 0 0 0 323

23 64 7 2 0 0 0 0 0 0 0 73

24 114 0 15 0 0 0 0 0 0 0 129

22 Low 296 208 120 0 59 161 10 0 0 0 854

23 123 8 0 0 0 0 0 0 0 0 131

24 239 13 59 12 0 0 0 0 0 0 323
120

Huntingdon Life Sciences

WAG0013

APPENDIX 5 Request ID: 257057

Bodyweight - individual values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Day Day Day Day Day Day Day Day Day Day Day Day

/Sex Number -5 -4 -3 -2 -1 1 2 3 4 5 6 7

1M 1 419 426 429 434 432 434 432 437 437 444 442 445

2 452 453 455 457 458 460 463 465 468 469 472 473

3 457 454 459 459 461 466 469 468 473 471 482 482

2M 4 369 375 380 384 393 396 384 385 393 385 387 391

5 395 400 403 407 417 424 409 413 416 411 410 408

6 410 419 430 434 445 451 425 437 451 446 445 446

3M 7 474 477 483 486 492 501 485 485 501 487 477 479

8 420 423 429 433 434 438 424 428 439 430 419 420

9 438 445 451 450 458 468 445 451 458 452 446 447

4M 10 467 465 468 468 473 471 442 449 461 449 449 441

11 429 431 436 432 437 427 423 413 430 417 394 377

12 452 454 458 459 459 461 449 444 465 448 434 432
121

Huntingdon Life Sciences

WAG0013

APPENDIX 5 - continued Request ID: 257057

Bodyweight - individual values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Day Day Day Day Day Day Day Day

/Sex Number 8 9 10 11 12 13 14 15

1M 1 443 439 448 450 448 451 448 454

2 477 479 483 488 487 489 490 490

3 481 484 485 489 489 491 492 497

2M 4 401 395 415 420 416 410 404 407

5 419 420 431 433 431 433 425 424

6 455 452 468 478 472 468 457 459

3M 7 477 473 502 513 513 502 491 502

8 422 426 440 454 446 450 444 444

9 444 448 469 482 469 467 454 463

4M 10 433 431 450 458 449 439 431 436

11 362 349 355 386 383 379 369 369

12 424 420 445 458 449 438 429 426
122

Huntingdon Life Sciences

WAG0013

APPENDIX 5 - continued Request ID: 257057

Bodyweight - individual values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Day Day Day Day Day Day Day Day Day Day Day Day

/Sex Number -5 -4 -3 -2 -1 1 2 3 4 5 6 7

1F 13 314 309 313 305 316 316 316 305 320 323 325 309

14 302 297 293 284 298 299 305 297 312 314 318 304

15 294 289 292 286 292 297 304 292 304 308 313 300

2F 16 242 244 246 250 247 249 246 252 255 252 258 243

17 230 237 240 239 236 247 233 233 234 229 238 238

18 248 249 243 253 256 254 247 250 257 252 253 260

3F 19 255 255 248 260 263 267 254 271 271 273 265 268

20 266 272 271 276 272 283 272 276 280 277 276 283

21 253 251 253 257 259 252 257 254 262 256 261 263

4F 22 252 260 264 260 256 266 259 261 272 262 259 271

23 272 281 285 278 275 280 263 270 288 273 263 265

24 263 255 264 267 267 260 259 258 269 259 261 262
123

Huntingdon Life Sciences

WAG0013

APPENDIX 5 - continued Request ID: 257057

Bodyweight - individual values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Animal Day Day Day Day Day Day Day Day

/Sex Number 8 9 10 11 12 13 14 15

1F 13 324 325 321 309 320 326 323 318

14 327 320 325 309 324 329 319 324

15 311 309 306 300 301 315 304 300

2F 16 252 256 279 281 264 267 263 265

17 238 241 247 256 256 253 247 251

18 262 262 269 270 269 275 273 274

3F 19 268 267 285 288 274 280 271 276

20 288 290 305 307 294 301 299 301

21 260 263 274 280 276 279 271 275

4F 22 265 270 281 277 272 269 267 267

23 241 245 271 285 277 258 242 258

24 258 260 276 285 283 258 270 271
124

Huntingdon Life Sciences

WAG0013

APPENDIX 6 Request ID: 257049

Food consumption - individual values (g/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Cage Day Day Day Day Day Day Day Day Day Day Day Day

/Sex Number -5 -4 -3 -2 -1 1 2 3 4 5 6 7

1M 1 31 30 31 31 28 28 29 30 31 30 29 30

2M 2 36 35 33 35 35 17 31 28 20 24 24 29

3M 3 34 33 35 35 35 17 27 29 22 21 22 21

4M 4 32 32 30 33 26 15 21 27 20 18 13 12
125

Huntingdon Life Sciences

WAG0013

APPENDIX 6 - continued Request ID: 257049

Food consumption - individual values (g/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Cage Day Day Day Day Day Day Day

/Sex Number 8 9 10 11 12 13 14

1M 1 30 31 32 28 31 30 30

2M 2 26 36 33 25 27 23 23

3M 3 24 37 36 31 30 30 27

4M 4 14 21 31 24 24 23 19
126

Huntingdon Life Sciences

WAG0013

APPENDIX 6 - continued Request ID: 257049

Food consumption - individual values (g/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Cage Day Day Day Day Day Day Day Day Day Day Day Day

/Sex Number -5 -4 -3 -2 -1 1 2 3 4 5 6 7

1F 5 25 26 20 29 21 25 20 29 27 27 19 30

2F 6 24 20 22 21 21 13 18 19 14 18 16 16

3F 7 30 19 26 26 27 12 21 24 18 20 19 17

4F 8 24 26 24 21 20 8 19 26 14 13 17 11
127

Huntingdon Life Sciences

WAG0013

APPENDIX 6 - continued Request ID: 257049

Food consumption - individual values (g/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group Cage Day Day Day Day Day Day Day

/Sex Number 8 9 10 11 12 13 14

1F 5 27 26 20 24 28 26 21

2F 6 19 25 25 18 22 15 16

3F 7 22 28 32 22 23 20 20

4F 8 13 24 27 17 17 11 14
128

Huntingdon Life Sciences

WAG0013

APPENDIX 7 Request ID: 257052

Organ weights - individual absolute values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal

Number

Terminal 

Bodyweight

Brain Heart Kidneys Liver Lungs + 

Bronchi

Spleen

1M 1 455 1.98 1.697 2.82 17.47 1.588 0.794

2 493 2.07 1.531 3.77 18.75 1.506 0.804

3 500 2.07 1.841 2.87 15.68 1.825 0.770

2M 4 411 1.98 1.758 3.20 14.48 1.667 0.738

5 424 2.03 1.269 3.43 14.41 1.522 0.563

6 460 2.00 1.342 3.71 18.26 1.713 0.687

3M 7 501 2.15 1.489 3.54 21.68 1.976 1.079

8 446 1.96 1.937 2.84 17.35 1.435 0.817

9 462 2.10 1.478 3.24 20.21 1.563 0.779

4M 10 437 2.19 1.770 3.25 17.65 1.987 0.669

11 370 1.97 1.373 2.34 15.29 1.731 0.573

12 427 2.11 1.563 3.32 15.51 1.472 0.667
129

Huntingdon Life Sciences

WAG0013

APPENDIX 7 - continued Request ID: 257052

Organ weights - individual absolute values (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Group

/Sex

Animal

Number

Terminal 

Bodyweight

Brain Heart Kidneys Liver Lungs + 

Bronchi

Spleen

1F 13 319 1.95 1.297 1.85 10.00 1.480 0.587

14 323 1.99 1.437 1.98 11.00 1.308 0.510

15 301 1.96 1.243 1.92 9.35 1.316 0.593

2F 16 267 1.93 1.051 2.03 10.71 1.295 0.544

17 252 1.68 0.835 1.96 8.71 1.095 0.572

18 274 1.91 0.895 1.69 8.47 1.181 0.414

3F 19 275 1.73 0.940 1.78 10.68 1.089 0.469

20 301 1.98 1.044 2.03 12.43 1.314 0.418

21 274 1.94 1.164 1.93 11.43 1.492 0.620

4F 22 267 1.84 1.055 1.67 10.10 1.318 0.505

23 257 1.86 1.043 1.77 9.82 1.449 0.500

24 271 1.90 0.988 1.71 10.31 1.216 0.448
130

Huntingdon Life Sciences

WAG0013

APPENDIX 8 Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 1 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 455.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED***

Lungs + Bronchi

 Alveolitis, Focal, Minimal

 Perivascular Inflammatory Cell Infiltrate, Minimal

 Arterial Mural Mineralisation, Focal, Minimal

Tracheal Bifurc.

 Inflammatory Cell Infiltrate in Lamina Propria, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 Larynx, Nasal turbinates, Trachea
131

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 2 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 493.4 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Lungs + Bronchi

 Alveolar Macrophages, Focal, Minimal

Skin

 Hair loss, Forelimb(s)

Tracheal Bifurc.

 Inflammatory Cell Infiltrate in Lamina Propria, Minimal

Trachea

 Inflammatory Cell Infiltrate - Lamina Propria, Focal, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 Larynx, Nasal turbinates
132

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 3 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 499.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Lungs + Bronchi

 Perivascular Inflammatory Cell Infiltrate, Minimal

 Arterial Mural Mineralisation, Focal, Minimal

Skin

 Hair loss, Forelimb(s)

The following examined tissues had no gross observations and no findings microscopically:

 Larynx, Nasal turbinates, Trachea, Tracheal Bifurc.
133

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 4 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 410.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Larynx

 Epithelial Keratinisation - Ventrolateral, Minimal

 Epithelial Hyperplasia - Arytenoid, Minimal
134

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 4 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 410.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Erosion, Focal, Minimal

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Minimal

 Olfactory Epithelium - Squamous Metaplasia, Focal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Focal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Focal, Minimal

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Slight

 Olfactory Nerve Fibres - Atrophy, Slight

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, Focal, 

Minimal

 Debris and Inflammatory Cells in Airway, Multifocal, Minimal
135

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 4 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 410.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Spleen

 Capsule thickened, Anterior pole, an area, 4x4mm.

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
136

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 5 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 424.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Lungs + Bronchi

 Pale area(s), Right, caudal lobe, Punctate, One, subpleural.

Larynx

 Submucosal Foreign Body Granulomata - Ventral, Focal, Minimal

 Squamous Metaplasia - Arytenoid, Slight
137

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 5 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 424.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Focal, 

Minimal

 Olfactory Epithelium - Squamous Metaplasia, Multifocal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Focal, Minimal

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Minimal

 Olfactory Nerve Fibres - Atrophy, Slight

 Respiratory Epithelium - Goblet Cell Hyperplasia, Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal

 Debris and Inflammatory Cells in Airway, Focal, Minimal

Spleen

 Capsule thickened, A few, areas, up to 7x5mm.
138

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 5 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 424.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
139

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 6 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 459.5 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED***

Larynx

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal

Nasal turbinates

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Focal, 

Minimal

 Olfactory Epithelium - Squamous Metaplasia, Focal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Focal, Minimal

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Minimal

 Olfactory Nerve Fibres - Atrophy, Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal

 Nasolacrimal Ducts - Inflammation, Unilateral, Minimal
140

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 6 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 459.5 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Tracheal Bifurc.

 Inflammatory Cell Infiltrate in Lamina Propria, Minimal

Trachea

 Inflammatory Cell Infiltrate - Lamina Propria, Focal, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 -
141

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 7 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 500.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Larynx

 Squamous Metaplasia - Ventral, Minimal

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal
142

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 7 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 500.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Erosion, Focal, Minimal

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Moderate

 Olfactory Epithelium - Squamous Metaplasia, Focal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Focal, Slight

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Moderate

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Slight

 Olfactory Nerve Fibres - Atrophy, Slight

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Multifocal, Moderate

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, 

Multifocal, Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal
143

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 7 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 500.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Skin

 Hair loss, Side(s) of face, diffuse.

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
144

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 8 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 445.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED***

Larynx

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal
145

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 8 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 445.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Focal, Slight

 Olfactory Epithelium - Squamous Metaplasia, Focal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Multifocal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Slight

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Slight

 Olfactory Nerve Fibres - Atrophy, Slight

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Focal, Moderate

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, 

Multifocal, Minimal

 Respiratory Epithelium - Goblet Cell Hyperplasia, Minimal

 Nasolacrimal Ducts - Inflammation, Minimal

 Debris and Inflammatory Cells in Airway, Multifocal, Minimal
146

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 8 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 445.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
147

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 9 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 461.8 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Lungs + Bronchi

 Dark area(s), Right, caudal lobe, One, subpleural, 1mm.

Larynx

 Submucosal Foreign Body Granulomata - Ventral, Focal, Minimal

 Squamous Metaplasia - Arytenoid, Minimal

 Epithelial Hyperplasia - Arytenoid, Minimal

 Inflammation, Multifocal, Minimal
148

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 9 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 461.8 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Multifocal, Slight

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Moderate

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Slight

 Olfactory Nerve Fibres - Atrophy, Slight

 Submucosal Foreign Body Granulomata - Dorsal, Focal, Minimal

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Focal, Moderate

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, 

Multifocal, Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal
149

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 9 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 461.8 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Skin

 Scab(s), Side(s) of face, Left, One, 1mm.

 Hair loss, Side(s) of face, diffuse.

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
150

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 10 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 437.0 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Kidneys

 Pale area(s), Left, Cranial pole, One, subcapsular, 4x2mm, near to pelvic 

region.

Lungs + Bronchi

 Arterial Mural Mineralisation, Focal, Minimal

Larynx

 Squamous Metaplasia - Ventral, Minimal

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Slight

 Inflammation, Multifocal, Minimal
151

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 10 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 437.0 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Erosion, Focal, Minimal

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Focal, 

Moderate

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Slight

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Slight

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Slight

 Submucosal Foreign Body Granulomata - Dorsal, Focal, Minimal

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Multifocal, Slight

 Respiratory Epithelium - Squamous Metaplasia, Focal, Minimal

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, 

Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Slight

 Nasolacrimal Ducts - Inflammation, Bilateral, Slight
152

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 10 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 437.0 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Stomach

 Forestomach roughened, Epithelial aspect

Tracheal Bifurc.

 Inflammatory Cell Infiltrate in Lamina Propria, Minimal

Trachea

 Epithelial Degeneration/Regeneration, Diffuse, Slight

 Inflammatory Cell Infiltrate - Lamina Propria, Diffuse, Minimal

Thymus

 Small

The following examined tissues had no gross observations and no findings microscopically:

 -
153

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 11 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 370.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Lungs + Bronchi Lungs + Bronchi

 Pale area(s), Left, Punctate, One, subpleural. Alveolitis, Focal, Minimal

 Alveolar Macrophages, Multifocal, Minimal

 Arterial Mural Mineralisation, Focal, Minimal

Larynx

 Epithelial Hyperplasia - Ventral, Minimal

 Epithelial Keratinisation - Ventrolateral, Minimal
154

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 11 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 370.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Necrosis, Focal, Minimal

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Slight

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Marked

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Marked

 Olfactory Nerve Fibres - Atrophy, Slight

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation,

Multifocal, Moderate

 Respiratory Epithelium - Squamous Metaplasia, Focal, Minimal

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, 

Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal
155

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 11 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 370.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Stomach

 Forestomach depression(s), Epithelial aspect, Two, crater-like, up to 6mm.

Trachea

 Epithelial Degeneration/Regeneration, Diffuse, Minimal

Thymus

 Small

The following examined tissues had no gross observations and no findings microscopically:

 Tracheal Bifurc.
156

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 12 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 427.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Kidneys

 Pelvic dilatation, Right

Larynx

 Squamous Metaplasia - Ventral, Minimal

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal
157

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 12 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 427.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Necrosis, Focal, Minimal

 Olfactory Epithelium - Erosion, Focal, Minimal

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Slight

 Olfactory Epithelium - Squamous Metaplasia, Focal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Slight

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Marked

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Marked

 Olfactory Nerve Fibres - Atrophy, Slight

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Multifocal, Moderate

 Respiratory Epithelium - Squamous Metaplasia, Focal, Minimal

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, 

Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal

 Nasolacrimal Ducts - Inflammation, Bilateral, Slight

 Debris and Inflammatory Cells in Airway, Focal, Slight
158

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 12 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 427.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Stomach

 Forestomach depression(s), Epithelial aspect, Punctate, One, crater-like.

Tracheal Bifurc.

 Inflammatory Cell Infiltrate in Lamina Propria, Minimal

Trachea

 Epithelial Degeneration/Regeneration, Diffuse, Slight

 Inflammatory Cell Infiltrate - Lamina Propria, Diffuse, Slight

Thymus

 Small

The following examined tissues had no gross observations and no findings microscopically:

 Lungs + Bronchi
159

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 13 Sex: Female Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 319.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED***

The following examined tissues had no gross observations and no findings microscopically:

 Larynx, Lungs + Bronchi, Nasal turbinates, Trachea, Tracheal Bifurc.
160

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 14 Sex: Female Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 322.8 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED***

Lungs + Bronchi

 Alveolar Macrophages, Focal, Minimal

 Alveolar Eosinophilic Crystals with Associated Inflammatory Cell Infiltrate, 

Focal, Minimal

 Arterial Mural Mineralisation, Focal, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 Larynx, Nasal turbinates, Trachea, Tracheal Bifurc.
161

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 15 Sex: Female Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 301.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED***

Lungs + Bronchi

 Alveolar Eosinophilic Crystals with Associated Inflammatory Cell Infiltrate, 

Focal, Minimal

 Arterial Mural Mineralisation, Focal, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 Larynx, Nasal turbinates, Trachea, Tracheal Bifurc.
162

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 16 Sex: Female Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 267.0 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED***

Larynx

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal
163

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 16 Sex: Female Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 267.0 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Squamous Metaplasia, Multifocal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Multifocal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Minimal

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Minimal

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, Focal, 

Minimal

 Nasolacrimal Ducts - Inflammation, Unilateral, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
164

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 17 Sex: Female Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 252.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

LN Bronchial

 Enlarged, 3mm.

Larynx

 Epithelial Keratinisation - Ventrolateral, Minimal
165

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 17 Sex: Female Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 252.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Erosion, Focal, Minimal

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Focal, Slight

 Olfactory Epithelium - Squamous Metaplasia, Multifocal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Multifocal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Minimal

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Minimal

 Respiratory Epithelium - Goblet Cell Hyperplasia, Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal
166

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 17 Sex: Female Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 252.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
167

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 18 Sex: Female Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 273.6 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

LN Mandibular

 Enlarged, Left, One, 13mm.

Lungs + Bronchi

 Pale area(s), Punctate, A few, subpleural.

Larynx

 Squamous Metaplasia - Arytenoid, Minimal
168

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 18 Sex: Female Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 273.6 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Focal, 

Minimal

 Olfactory Epithelium - Squamous Metaplasia, Multifocal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Multifocal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Minimal

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Slight

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Focal, Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal
169

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 18 Sex: Female Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 25-Nov-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 273.6 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
170

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 19 Sex: Female Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 275.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED***

Larynx

 Squamous Metaplasia - Ventral, Minimal

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal
171

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 19 Sex: Female Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 275.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Slight

 Olfactory Epithelium - Squamous Metaplasia, Multifocal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Focal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Moderate

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Slight

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Multifocal, Moderate

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, Focal, 

Minimal

 Debris and Inflammatory Cells in Airway, Multifocal, Minimal
172

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 19 Sex: Female Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 275.2 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Trachea

 Inflammatory Cell Infiltrate - Lamina Propria, Diffuse, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 Tracheal Bifurc.
173

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 20 Sex: Female Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 300.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED***

Larynx

 Squamous Metaplasia - Ventral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal
174

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 20 Sex: Female Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 300.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Slight

 Olfactory Epithelium - Squamous Metaplasia, Focal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Multifocal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Moderate

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Slight

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Multifocal, Moderate

 Transitional Epithelium - Disorganisation/Regeneration, Minimal
175

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 20 Sex: Female Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 300.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
176

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 21 Sex: Female Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 274.4 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED***

Larynx

 Squamous Metaplasia - Ventral, Minimal

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal
177

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 21 Sex: Female Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 274.4 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Necrosis, Focal, Minimal

 Olfactory Epithelium - Erosion, Focal, Minimal

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Slight

 Olfactory Epithelium - Squamous Metaplasia, Multifocal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Multifocal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Moderate

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Moderate

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Multifocal, Moderate

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, 

Multifocal, Slight

 Respiratory Epithelium - Goblet Cell Hyperplasia, Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal

 Debris and Inflammatory Cells in Airway, Focal, Minimal
178

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 21 Sex: Female Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 02-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 274.4 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Tracheal Bifurc.

 Inflammatory Cell Infiltrate in Lamina Propria, Minimal

Trachea

 Epithelial Degeneration/Regeneration, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 -
179

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 22 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 266.8 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Lungs + Bronchi

 Alveolar Macrophages, Focal, Minimal

Larynx

 Squamous Metaplasia - Ventral, Minimal

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal
180

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 22 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 266.8 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Slight

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Moderate

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Moderate

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Multifocal, Moderate

 Respiratory Epithelium - Squamous Metaplasia, Focal, Minimal

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, 

Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Minimal

 Nasolacrimal Ducts - Inflammation, Bilateral, Minimal
181

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 22 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 266.8 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Stomach

 Forestomach roughened, Epithelial aspect, an area, 6mm.

Thymus

 Small

The following examined tissues had no gross observations and no findings microscopically:

 Trachea, Tracheal Bifurc.
182

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 23 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 256.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Lungs + Bronchi

 Alveolar Macrophages, Diffuse, Minimal

 Arterial Mural Mineralisation, Focal, Minimal

Larynx

 Squamous Metaplasia - Ventral, Minimal

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal

 Epithelial Hyperplasia - Arytenoid, Minimal

 Inflammation, Multifocal, Slight
183

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 23 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 256.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Erosion, Focal, Minimal

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Moderate

 Olfactory Epithelium - Squamous Metaplasia, Focal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Focal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Moderate

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Slight

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Multifocal, Moderate

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, 

Minimal

 Transitional Epithelium - Disorganisation/Regeneration, Slight
184

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 23 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 256.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Tracheal Bifurc.

 Inflammatory Cell Infiltrate in Lamina Propria, Minimal

Trachea

 Epithelial Degeneration/Regeneration, Diffuse, Minimal

 Inflammatory Cell Infiltrate - Lamina Propria, Diffuse, Slight

Thymus

 Small

The following examined tissues had no gross observations and no findings microscopically:

 -
185

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 24 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 271.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Lungs + Bronchi

 Alveolar Macrophages, Multifocal, Minimal

Larynx

 Squamous Metaplasia - Ventral, Minimal

 Epithelial Keratinisation - Ventrolateral, Minimal

 Squamous Metaplasia - Arytenoid, Minimal

 Epithelial Hyperplasia - Arytenoid, Minimal
186

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 24 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 271.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Nasal turbinates

 Olfactory Epithelium - Degeneration/Atrophy/Disorganisation, Multifocal, 

Moderate

 Olfactory Epithelium - Squamous Metaplasia, Focal, Minimal

 Olfactory Epithelium - Respiratory Metaplasia With or Without Associated 

Inflammatory Cell Infiltrate, Focal, Minimal

 Olfactory Epithelium - Fibrosis With or Without Associated Cartilage 

Formation in Dorsal Lamina Propria, Moderate

 Olfactory Epithelium - Bowman's Gland Atrophy in Dorsal Lamina Propria, 

Moderate

 Olfactory Nerve Fibres - Atrophy, Slight

 Fusion of Dorsal Turbinate With Associated Epithelial Diverticula Formation, 

Multifocal, Moderate

 Respiratory Epithelium - Disorganisation/Degeneration/ Regeneration, Slight

 Transitional Epithelium - Disorganisation/Regeneration, Minimal

 Nasolacrimal Ducts - Epithelial Degeneration, Bilateral, Minimal

 Nasolacrimal Ducts - Inflammation, Unilateral, Minimal

 Debris and Inflammatory Cells in Airway, Focal, Minimal
187

Huntingdon Life Sciences

WAG0013

APPENDIX 8 - continued Request ID: 264438

Macropathology and histopathology - individual findings

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure (ppm) : 0 4721 8901 19098

Animal Number: 24 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice

Date of Death: 09-Dec-11 Day of Death: 15 Week of Death: 3 Subgroup: 1 Terminal Bodyweight: 271.3 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

continued from previous page...

Stomach

 Forestomach roughened, Epithelial aspect

Trachea

 Epithelial Degeneration/Regeneration, Diffuse, Minimal

Thymus

 Small

The following examined tissues had no gross observations and no findings microscopically:

 Tracheal Bifurc.
Huntingdon Life Sciences

WAG0013

188

Annex 1 Administration of 2-bromo-3,3,3-trifluoropropene to rats
Huntingdon Life Sciences

WAG0013

189

Working for a better future

Aerosol Technology Report

HLS study number: WAG0013

Author A Allen

Version ID: Audited Draft 1

Issue date:
Huntingdon Life Sciences

WAG0013

190 

Table of Contents 

Signature Page ................................................................................................................... 191

1. Pre-study Atmosphere Characterization ..................................................................... 192

2. Test Article Handling and Processing for Inhalation Generation ................................. 192

3. Exposure System ........................................................................................................ 192

4. Administration ............................................................................................................. 193

5. Atmosphere Analysis .................................................................................................. 193

5.1 Nominal Concentration ......................................................................................... 193

5.2 Nominal Concentration ......................................................................................... 194

5.3 Chamber Temperature ......................................................................................... 194

5.4 Chamber Humidity ................................................................................................ 194

6. Data and Calculations ................................................................................................. 195

7. Results and Discussion ............................................................................................... 195

7.1 Aerosol Concentrations ............................................................................. 195

List of Tables 

Table 1 Chamber atmosphere concentration of 2-bromo-3,3,3-trifluoropropene - 

individual exposure values for Group 1 ....................................................... 196

Table 2 Chamber atmosphere concentration of 2-bromo-3,3,3-trifluoropropene – daily 

means and individual exposure values for Group 2 .................................... 197

Table 3 Chamber atmosphere concentration of 2-bromo-3,3,3-trifluoropropene - daily 

means and individual exposure values for Group 3 .................................... 198

Table 4 Chamber atmosphere concentration of 2-bromo-3,3,3-trifluoropropene - daily 

means and individual exposure values for Group 4 .................................... 199

Table 5 Chamber Temperatures - daily mean .......................................................... 200

Table 6 Chamber Humidity - daily mean .................................................................. 201

Table 7 Nominal Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene - individual exposure values for Group 2 ... 202

Table 8 Nominal Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene - individual exposure values for Group 3 ... 203

Table 9 Nominal Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene - individual exposure values for Group 4 ... 204

List of Figures 

Figure 1 Schematic of Inhalation Exposure System .................................................. 205

Huntingdon Life Sciences

WAG0013

192

1. Pre-study Atmosphere Characterization

Inhalation exposure system characterized including determinations of:

Achieved vapour concentration;

Spatial distribution around the chamber. 

2. Test Article Handling and Processing for Inhalation 

Generation

As supplied, 2-Bromo-3,3,3-trifluoro-1-propene was easily vaporised using warmed air 

through a sintered glass vaporiser.

3. Exposure System

A schematic of the exposure system is presented in Figure 1. Components of the exposure 

system:

Directed flow Whole body exposure chamber:

0.75m3 chamber of stainless–steel and glass construction

Animal Restraint:

Wire mesh modular caging

Aerosol Generator:

Sintered glass vaporiser for Groups 2 to 4

Liquid reservoir and feed rate used to alter concentration

Group 2 – 29L/min

Groups 3 and 4 – 49L/min

Diluent Airflow:

From in-house compressed air system – breathing quality

Group 1 – 149L/min

Group 2 – 120L/min

Groups 3 and 4 – 100L/min

Extract Airflow:

Drawn by a fan extract system

Drawn from base of exposure system

Controlled by a gate clamp situated at the rear of the chamber

Airflow Monitoring:

In-line flowmeters monitored continuously for generation and diluent 

airflows

Chamber pressure differential monitored by Magnahelic set at 1 to 

4mm/H2O to ensure a slight negative pressure within the 

chamber~150L/min.
Huntingdon Life Sciences

WAG0013

193

4. Administration

Animals exposed for 6 hours per day, 5 days a week for 2 weeks

Exposure system operational parameters monitored continuously

Airflows documented initially and monitored during exposure

5. Atmosphere Analysis

5.1 Nominal Concentration

Aerosol samples collected as follows:

Sample frequency – minimum of 3 samples/group/exposure 

Sample location – representative animal level

Sample position remained constant

Sample analysis

Infra-red spectrophotometer
Huntingdon Life Sciences

WAG0013

194

Flow diagram of the analytical method for the analysis of 

2-bromo-3,3,3-trifluoropropene in test atmosphere samples

Sample Assay

Test atmosphere sample (Gas bag or chamber)



Sample directly using the IR spectrophotometer



Perform a single measure on the IR 

spectrophotometer 

Calibration

Prepare a series of calibration standards by direct 

addition of a suitable volume of test item directly into 

gas bags containing a suitable volume of air



Select and prepare the appropriate calibration range 

for the expected concentration of the test 

atmosphere samples to be analysed



Prepare a set of calibration standards for low range 

analysis at nominal concentrations of 1167, 2334, 

3501, 4668, 7002 and 9336 ppm



Prepare a set of calibration standards for inter range 

analysis at nominal concentrations of 2334, 3501, 

4668, 9336, 14004 and 18672 ppm



Prepare a set of calibration standards for high range 

analysis at nominal concentrations of 4668, 9336, 

14004, 18672, 23340 and 28008 ppm



Measure the required calibration range standards 

using the IR spectrophotometer for calibration

IR conditions

Instrument: MIRAN SapphIRe model ML

Path length: Short (0.5m)

Wavelength: 10.699 μm (low calibration range)

12.026 μm (inter and high calibration range)

The IR spectrophotometer was calibrated using external standards. Absorbance 

measurements acquired by the IR spectrophotometer were used to calculate analysed 

concentration (ppm) using a 2nd order fit subjected to least squares regression analysis.

5.2 Nominal Concentration

Monitored and summarised in Tables 7 to 9.

5.3 Chamber Temperature 

Monitored using a liquid in glass thermometer, daily means presented in Table 5. 

5.4 Chamber Humidity 

Monitored using wet and dry liquid in glass thermometers, with humidity calculated using 

proprietary tables, daily means presented in Table 6.
Huntingdon Life Sciences

WAG0013

195

6. Data and Calculations

To minimize the cumulative errors from repeated rounding of numbers, data in this report has 

been calculated continuously using unrounded numbers and only rounded for printing. 

Consequently, presented data may include rounding errors in the last significant figure.

All data not presented in this report is retained in the study data files and archives. 

7. Results and Discussion

7.1 Aerosol Concentrations

Individual exposure concentrations and daily means are presented in Tables 1 to 4 and 

summarized below:

Group Concentration (ppm)

Target Mean

1 0

2 5000 4721

3 10000 8901

4 20000 19098

Additional samples were taken for confirmation of atypical results and after minor feed rate 

adjustments.

All groups were relatively close to target.
Huntingdon Life Sciences

WAG0013

196

Table 1 Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene - individual exposure values for 

Group 1

Exposure

No.

Chamber concentration (ppm)

Sample 1 Sample 2 Sample 3

1 ND ND ND

2 ND ND ND

3 ND ND ND

4

1 ND ND ND

5 ND ND ND

6

1 ND ND ND

7 ND ND ND

8

1 ND ND ND

9 ND ND ND

10

1 ND ND ND

ND No test item detected

1 Only one standard curve was used for calibration due to insufficient volume remaining for 

measurement
Huntingdon Life Sciences

WAG0013

197

Table 2 Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene – daily means and individual 

exposure values for Group 2

Exposure

No.

Chamber concentration (ppm)

Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6 Daily 

mean

1 5296 5108 4751 4588 4936

2 5581 4383 4267 5123 4423 4912 4782

3

1

5546 4760 4878 5061

4 4062 5661 4859 3554 4534

5 6113 5613 4530 3170 2946 4474

6 6611 4160 4732 5758 5315

7 5789 4101 4851 4914

8

1

5978 5008 4357 3851 4736 4786

9

1

4380 4298 4013 4230

10 4523 4021 3981 4175

Mean 24721

sd 364

CV (%) 7.7

1 Only one standard curve was used for calibration due to insufficient volume remaining for 

measurement

2 Mean calculated from the individual mean values

sd Standard deviation

CV (%) Coefficient of variation (sd × 100/mean)
Huntingdon Life Sciences

WAG0013

198

Table 3 Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene - daily means and individual 

exposure values for Group 3

Exposure

No.

Chamber concentration (ppm)

Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6 Daily 

mean

1 9817 6681 10288 10080 9217

2 6902 4723 11694 7773

3

1

11319 7964 7288 15143 7056 10692 9910

4

1

7524 10077 8648 8750

5 7860 9833 9563 9085

6 9304 11418 9570 10097

7

2

8247 8593 8017

8

1

7130 7928 7977 7678

9

1

9170 8419 7304 8298

10 8947 9003 9939 9296

Mean 38901

sd 858

CV (%) 9.6

1 Only one standard curve was used for calibration due to insufficient volume remaining for 

measurement 

2 Results from Exposure 7 have been excluded from the mean, due to the standard measurements falling 

outside acceptance criteria

3 Mean calculated from the individual mean values

sd Standard deviation

CV (%) Coefficient of variation (sd × 100/mean)
Huntingdon Life Sciences

WAG0013

199

Table 4 Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene - daily means and individual 

exposure values for Group 4

Exposure

No.

Aerosol concentration (ppm)

Sample 1 Sample 2 Sample 3 Sample 4 Daily 

mean

1 20053 17539 19272 18955

2

1

18656 17536 18093

3

2

18487 18472 18193 18384

4

2

10617 20131 18076 19121 16986

5 20434 19500 20019 19984

6

2

18547 20381 17974 18967

7 16028 18501 19457 17995

8

2

23547 17121 19444 20037

9 20840 23887 20573 21767

10

2

13683 25746 20232 15580 18810

Mean 319098

sd 1374

CV (%) 7.2

1 Results from Exposure 2 have been excluded from the mean, due to the standard measurements falling 

outside acceptance criteria

2 Only one standard curve was used for calibration due to insufficient volume remaining for 

measurement

3 Mean calculated from the individual mean values

sd Standard deviation

CV (%) Coefficient of variation (sd × 100/mean)
Huntingdon Life Sciences

WAG0013

200

Table 5 Chamber Temperatures - daily mean

Exposure

No.

Chamber Temperature (°C)

Group 1 Group 2 Group 3 Group 4

1 21.0 20.7 20.5 20.8

2 21.0 20.8 20.6 20.4

3 20.6 20.5 20.4 20.0

4 21.0 20.7 20.5 20.2

5 21.0 20.5 20.6 20.1

6 20.9 20.6 20.7 20.0

7 21.0 20.6 20.7 20.0

8 21.0 20.5 20.0 20.0

9 21.0 20.5 20.2 20.2

10 21.1 20.6 20.4 20.4
Huntingdon Life Sciences

WAG0013

201

Table 6 Chamber Humidity - daily mean

Exposure

No.

Chamber Temperature (%RH)

Group 1 Group 2 Group 3 Group 4

1 28 34 28 31

2 29 25 25 32

3 29 29 26 32

4 33 27 27 34

5 26 26 26 32

6 22 30 29 35

7 23 28 26 32

8 24 28 28 32

9 23 30 28 31

10 22 29 25 31
Huntingdon Life Sciences

WAG0013

202

Table 7 Nominal Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene - individual exposure values for 

Group 2

Day

Barometric 

Pressure 

P(mmHg)

Test 

Compound

usage TCU

(g)

Volume of 

Test

Compound 

V (L)

Analysed 

Conc.(ppm)

Nominal 

Conc.(ppm)

A/N

1

ratio%

1 764 1980 271.3 4936 4999 98.7

2 769 2020 275.1 4782 5068 94.4

3 766 2060 281.4 5061 5184 97.6

4 786 1610 214.5 4534 3956 115

5 757 1760 243.3 4474 4485 99.8

6 763 2050 281.2 5315 5180 103

7 761 2050 281.9 4914 5194 94.6

8 764 1960 268.4 4786 4946 96.8

9 770 1700 231.0 4230 4259 99.3

10 770 1550 210.7 4175 3887 107

1 Analysed/Nominal (A/N) ratio (%) = Analysed concentration (ppm) / Nominal Concentration 

(ppm)*100

 

P(mmHg)

760

MW (g/mol)

TCU (g) R T( C) 273 )( 

LV 

(F D) V(L)

6

)( 10

NominalConc (ppm)







LV

V =Volume of test article 

R =Gas constant (0.08206Latm/mol°K)

MW =Molecular weight (174.95g/mol)

F =Chamber airflow (150L/min)

D =Duration 6 Hours (360mins)
Huntingdon Life Sciences

WAG0013

203

Table 8 Nominal Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene - individual exposure values for 

Group 3

Day Barometric 

Pressure 

P(mmHg)

Test 

Compound

usage TCU

(g)

Volume of 

Test

Compound 

V (L)

Analysed 

Conc.(ppm)

Nominal 

Conc.(ppm)

A/N2

ratio%

1 763 3470 475.9 9217 8736 106

2 761 3330 457.9 7773 8409 92.4

3 764 3470 475.0 9910 8720 114

4 770 3440 467.4 8750 8582 102

5 770 3490 474.3 9085 8708 104

6 767 3660 499.5 10097 9166 110

7 768 2980 406.2 7467 1

8 753 3250 450.9 7678 8280 92.7

9 765 3350 457.6 8298 8403 98.7

10 756 3630 502.2 9296 9214 101

1 No value due to analysed value excluded

2 Analysed/Nominal (A/N) ratio (%) = Analysed concentration (ppm) / Nominal Concentration 

(ppm)*100

 

P(mmHg)

760

MW (g/mol)

TCU(g) R T( C) 273 )( 



LV 

(F D) V(L)

6

)( 10 NominalConc.(ppm)







LV

V =Volume of test article 

R =Gas constant (0.08206Latm/mol°K)

MW =Molecular weight (174.95g/mol)

F =Chamber airflow (150L/min)

D =Duration 6 Hours (360mins)
Huntingdon Life Sciences

WAG0013

204

Table 9 Nominal Chamber atmosphere concentration of 

2-bromo-3,3,3-trifluoropropene - individual exposure values for 

Group 4

Day Barometric 

Pressure 

P(mmHg)

Test 

Compound

usage TCU

(g)

Volume of 

Test

Compound 

V (L)

Analysed 

Conc.(ppm)

Nominal 

Conc.(ppm)

A/N2

ratio%

1 767 7120 972.4 18955 17689 107

2 768 6270 853.9 15568 1

3 753 6580 912.7 18384 16621 111

4 765 6100 833.3 16986 15197 112

5 756 6780 937.0 19984 17055 117

6 761 7060 969.0 18967 17628 108

7 754 7340 1016.9 17995 18484 97.4

8 754 7030 974.0 20037 17717 113

9 753 7400 1027.1 21767 18666 117

10 754 7220 1001.5 18810 18208 103

1 No value due to analysed value excluded

2 Analysed/Nominal (A/N) ratio (%) = Analysed concentration (ppm) / Nominal Concentration 

(ppm)*100

(F D) V(L)

6

)( 10 NominalConc.(ppm)







LV

V =Volume of test article 

R =Gas constant (0.08206Latm/mol°K)

MW =Molecular weight (174.95g/mol)

F =Chamber airflow (150L/min)

D =Duration 6 Hours (360mins)

 

P(mmHg)

760

MW (g/mol)

TCU(g) R T( C) 273 )( 

LV 
Huntingdon Life Sciences

WAG0013

205

Figure 1 Schematic of Inhalation Exposure System

A

B

D

F

C

E

F

H

I

J

K

Front view Side view

L

Generation Trolley

G

Key

A Whole body chamber G Load cells

B 76.2mm test material delivery tubing H Air-line coil

C Elutriator I Sintered glass vaporiser

D Secondary generation trolley J Water bath

E Magnahelix K Generation flowmeters

F Pressure Vessels L Control valves
Huntingdon Life Sciences

WAG0013

206

Annex 2 Pathology report
207
Huntingdon Life Sciences

WAG0013

 Issue No: 2 

Page 2 of 10 

1. Introduction 

1.1 Objective 

Assessment of local and systemic toxic potential in a 14 day inhalation study in rats to select 

suitable exposure levels for a subsequent 13-week inhalation toxicity study. 

Group Treatment 

Achieved 

exposure level 

(ppm) 

Number of animals 

Male Female 

 

1 Control 0 3 3 

2 2-bromo-3,3,3-trifluoropropene 4721 3 3 

3 2-bromo-3,3,3-trifluoropropene 8901 3 3 

4 2-bromo-3,3,3-trifluoropropene 19098 3 3 

 

2. Results 

2.1 Decedents 

There were no unscheduled deaths during the study. 

2.2 Macroscopic Pathology 

2.2.1 Animals killed after 2 weeks of treatment 

The macroscopic examination performed after 2 weeks of treatment revealed the following 

intergroup differences: 

Thymus 

A reduction in the size of the thymus was noted in animals treated with 19098 ppm. 

Summary of findings in the thymus for animals killed after 2 weeks of treatment 

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F 

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098 

 

Small 0 0 0 3 0 0 0 3 

Number of animals examined 3 3 3 3 3 3 3 3 

 

208
Huntingdon Life Sciences

WAG0013

 Issue No: 2 

Page 3 of 10 

Stomach 

Depressions or roughening of the stomach was observed in animals treated with 19098 ppm. 

Summary of findings in the stomach for animals killed after 2 weeks of treatment 

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F 

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098 

 

Forestomach Depression(s) 0 0 0 2 0 0 0 0 

Forestomach Roughened 0 0 0 1 0 0 0 2 

Number of animals examined 3 3 3 3 3 3 3 3 

 

The nature and incidence of all other findings were consistent with the commonly seen 

background of macroscopic changes in rats at these laboratories. 

2.3 Microscopic Pathology 

2.3.1 Animals killed after 2 weeks of treatment 

Treatment related findings 

Changes related to treatment with 2-bromo-3,3,3-trifluoropropene were seen in the nasal 

turbinates, larynx and trachea. 

Nasal turbinates 

In the olfactory epithelium of male and female animals exposed to concentrations of 4721, 

8901 or 19098 ppm, the test article-related findings seen included the following. 

Minimal to marked fibrosis with or without associated cartilage formation in the dorsal 

lamina propria. 

Minimal to marked atrophy of the Bowman’s glands in the dorsal lamina propria. 

Minimal to moderate atrophy of olfactory nerve fibres. 

Minimal epithelial necrosis was seen in two male animals exposed to 19098 ppm and one 

female exposed to 8901 ppm. 

Minimal erosion of the epithelium. 

Minimal to moderate degeneration/atrophy/disorganisation of the epithelium. 

Minimal squamous metaplasia of the epithelium. 

Minimal or slight respiratory metaplasia of the epithelium with or without associated 

inflammatory cell infiltrate. 

209
Huntingdon Life Sciences

WAG0013

 Issue No: 2 

Page 4 of 10 

In the turbinates, minimal to moderate fusion of dorsal turbinates with associated epithelial 

diverticula formation was seen in male and female animals exposed to 4721, 8901 or 

19098 ppm. 

In the respiratory epithelium of male and female animals exposed to 4721, 8901 or 

19098 ppm, the test article-related findings seen included the following. 

Minimal or slight disorganisation/degeneration/regeneration of the epithelium. 

Minimal squamous metaplasia of the epithelium was only seen in animals exposed to 

19098 ppm. 

In the transitional epithelium, minimal or slight disorganisation/regeneration of the 

epithelium was seen in male and female animals exposed to 4721, 8901 or 19098 ppm. 

In the nasolacrimal ducts, minimal epithelial degeneration was seen in one female exposed to 

19098 ppm and minimal or slight inflammation was seen in animals exposed to 4721, 8901 or 

19098 ppm. In the airway, minimal or slight debris and inflammatory cells were also seen in 

animals exposed to 4721, 8901 or 19098 ppm. 

210
Huntingdon Life Sciences

WAG0013

 Issue No: 2 

Page 5 of 10 

Summary of treatment related findings in the nasal turbinates for animals killed after 2 weeks of 

treatment 

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F 

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098 

 

Olfactory epithelium 

Fibrosis with or without associated 

cartilage formation in dorsal lamina 

propria 

Minimal 

Slight 

Moderate 

Marked 

0 

0 

0 

0 

2 

0 

0 

0 

0 

1 

2 

0 

0 

1 

0 

2 

0 

0 

0 

0 

3 

0 

0 

0 

0 

0 

3 

0 

0 

0 

3 

0 

Total 0 2 3 3 0 3 3 3 

 

Bowman’s gland atrophy in dorsal 

lamina propria 

Minimal 

Slight 

Moderate 

Marked 

0 

0 

0 

0 

2 

1 

0 

0 

0 

3 

0 

0 

0 

0 

1 

2 

0 

0 

0 

0 

0 

0 

3 

0 

0 

0 

3 

0 

0 

0 

3 

0 

Total 0 3 3 3 0 3 3 3 

 

Olfactory nerve fibres - atrophy 

Minimal 

Slight 

Moderate 

0 

0 

0 

1 

2 

0 

0 

3 

0 

0 

3 

0 

0 

0 

0 

2 

1 

0 

0 

2 

1 

0 

2 

1 

Total 0 3 3 3 0 3 3 3 

 

Necrosis 

Minimal 0 0 0 2 0 0 1 0 

Total 0 0 0 2 0 0 1 0 

 

Erosion 

Minimal 0 1 1 2 0 1 1 1 

Total 0 1 1 2 0 1 1 1 

 

Degeneration/atrophy/disorganisation 

Minimal 

Slight 

Moderate 

0 

0 

0 

3 

0 

0 

1 

1 

1 

0 

2 

1 

0 

0 

0 

1 

1 

0 

0 

3 

0 

0 

1 

2 

Total 0 3 3 3 0 2 3 3 

 

Squamous metaplasia 

Minimal 0 3 2 1 0 3 3 2 

Total 0 3 2 1 0 3 3 2 

 

Respiratory metaplasia with or 

without associated inflammatory cell 

infiltrate 

Minimal 

Slight 

0 

0 

2 

0 

1 

2 

1 

2 

0 

0 

3 

0 

3 

0 

2 

0 

Total 0 2 3 3 0 3 3 2 

 

211
Huntingdon Life Sciences

WAG0013

 Issue No: 2 

Page 6 of 10 

Summary of treatment related findings in the nasal turbinates for animals killed after 2 weeks of 

treatment 

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F 

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098 

 

Turbinates 

Fusion of dorsal turbinate with 

associated epithelial diverticula 

formation 

Minimal 

Slight 

Moderate 

0 

0 

0 

0 

0 

0 

0 

0 

3 

0 

1 

2 

0 

0 

0 

1 

0 

0 

0 

0 

3 

0 

0 

3 

Total 0 0 3 3 0 1 3 3 

 

Respiratory epithelium 

Disorganisation/Degeneration/ 

Regeneration 

Minimal 

Slight 

0 

0 

1 

0 

3 

0 

3 

0 

0 

0 

1 

0 

1 

1 

2 

1 

Total 0 1 3 3 0 1 2 3 

 

Squamous metaplasia 

Minimal 0 0 0 3 0 0 0 1 

Total 0 0 0 3 0 0 0 1 

 

Transitional epithelium 

Disorganisation/Regeneration 

Minimal 

Slight 

0 

0 

2 

0 

2 

0 

2 

1 

0 

0 

2 

0 

2 

0 

2 

1 

Total 0 2 2 3 0 2 2 3 

 

Nasolacrimal ducts 

Epithelial degeneration 

Minimal 0 0 0 0 0 0 0 1 

Total 0 0 0 0 0 0 0 1 

 

Inflammation 

Minimal 

Slight 

0 

0 

1 

0 

1 

0 

0 

2 

0 

0 

1 

0 

0 

0 

2 

0 

Total 0 1 1 2 0 1 0 2 

 

Airway 

Debris and inflammatory cells 

Minimal 

Slight 

0 

0 

2 

0 

1 

0 

0 

1 

0 

0 

0. 

0 

2 

0 

1 

0 

Total 0 2 1 1 0 0 2 1 

 

Number of tissues examined 3 3 3 3 3 3 3 3 

212
Huntingdon Life Sciences

WAG0013

 Issue No: 2 

Page 7 of 10 

Larynx 

In the ventral larynx, minimal squamous metaplasia was seen in male and female animals 

exposed to 8901 or 19098 ppm and minimal epithelial hyperplasia was seen in one male 

exposed to 19098 ppm. 

In the ventrolateral larynx, minimal epithelial keratinisation was seen in male and female 

animals of all groups exposed to 2-bromo-3,3,3-trifluoropropene. 

In the arytenoids, minimal or slight squamous metaplasia and minimal epithelial hyperplasia 

were seen in animals of all test article groups. 

Minimal or slight inflammation was seen in one male and one female exposed to 19098 ppm 

and one male exposed to 8901 ppm.

Summary of treatment related findings in the larynx for animals killed after 2 weeks of treatment 

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F 

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098 

 

Squamous metaplasia - ventral 

Minimal 0 0 1 2 0 0 3 3 

Total 0 0 1 2 0 0 3 3 

 

Epithelial hyperplasia - ventral 

Minimal 0 0 0 1 0 0 0 0 

Total 0 0 0 1 0 0 0 0 

 

Epithelial keratinisation - 

ventrolateral 

Minimal 0 2 2 3 0 2 2 3 

Total 0 2 2 3 0 2 2 3 

 

Squamous metaplasia - arytenoid 

Minimal 

Slight 

0 

0 

1 

1 

3 

0 

1 

1 

0 

0 

2 

0 

3 

0 

3 

0 

Total 0 2 3 2 0 2 3 3 

 

Epithelial hyperplasia - arytenoid 

Minimal 0 1 1 0 0 0 0 2 

Total 0 1 1 0 0 0 0 2 

 

Inflammation 

Minimal 

Slight 

0 

0 

0 

0 

1 

0 

1 

0 

0 

0 

0 

0 

0 

0 

0 

1 

Total 0 0 1 1 0 0 0 1 

 

Number of tissues examined 3 3 3 3 3 3 3 3 

213
Huntingdon Life Sciences

WAG0013

 Issue No: 2 

Page 8 of 10 

Trachea 

Minimal or slight epithelial degeneration/regeneration was seen in male and female animals 

exposed to 19098 ppm and one female exposed to 8901 ppm. Minimal or slight 

inflammatory cell infiltrate in the lamina propria was seen at a higher incidence and/or degree 

in animals exposed to 19098 ppm, than in control animals.

Summary of treatment related findings in the trachea for animals killed after 2 weeks of treatment 

Group/sex 1M 2M 3M 4M 1F 2F 3F 4F 

Achieved Exposure Level (ppm) 0 4721 8901 19098 0 4721 8901 19098 

 

Epithelial 

degeneration/regeneration 

Minimal 

Slight 

0 

0 

0 

0 

0 

0 

1 

2 

0 

0 

0 

0 

1 

0 

2 

0 

Total 0 0 0 3 0 0 1 2 

 

Inflammatory cell infiltrate – 

lamina propria 

Minimal 

Slight 

1 

0 

1 

0 

0 

0 

1 

1 

0 

0 

0 

0 

1 

0 

0 

1 

Total 1 1 0 2 0 0 1 1 

 

Number of tissues examined 3 3 3 3 3 3 3 3 

214
Huntingdon Life Sciences

WAG0013

 Issue No: 2 

Page 9 of 10 

3. Discussion 

In the nasal turbinates, the findings related to treatment were generally seen in the tissues of 

the dorsal half of the nasal cavity, with the severity of findings increasing in a rostral to 

caudal direction and the olfactory epithelium being the most extensively affected. 

In the region of the olfactory epithelium, fibrosis with or without associated cartilage 

formation, atrophy of the Bowman’s glands and atrophy of olfactory nerve fibres were seen 

in the dorsal lamina propria and erosion of the epithelium were seen in animals of all groups 

exposed to 2-bromo-3,3,3-trifluoropropene. Necrosis of the olfactory epithelium was seen in 

animals exposed to 8901 or 19098 ppm. The fibrosis with or without associated cartilage 

formation is considered a protective response to irritation by the test substance, which would 

have caused the necrosis and erosion, with the atrophy of the Bowman’s glands and olfactory 

nerves being either a direct effect of the irritation or a consequence of the epithelial 

degeneration. 

The fusion of dorsal turbinates with associated epithelial diverticula formation seen in 

animals of all test article groups, was considered a result of damage to the epithelium 

overlying the turbinates and dorsal aspect of the nasal cavity with subsequent fusion of the 

turbinates and formation of epithelial diverticula. 

In the various epithelia of the nasal cavity (olfactory, respiratory and transitional) of animals 

of all test article groups similar findings were seen. These included metaplasia and 

combinations of disorganisation, atrophy, degeneration and regeneration. The metaplasia 

was an adaptive change to produce a type of epithelium which was more resistant to the 

irritation. The combinations of disorganisation, atrophy and degeneration were due to the 

irritation of the epithelium and the regeneration was part of the tissue repair process. 

In the nasolacrimal ducts, inflammation was seen in animals of all test article groups. 

Although epithelial degeneration was seen in only one female exposed to 19098 ppm, both 

the inflammation and epithelial degeneration are considered due to the irritancy of the test 

substance. 

In the airway, debris and inflammatory cells were seen in of all test article groups and are 

considered a consequence of the other changes in the nasal cavity. 

In general, the changes seen in the nasal turbinates of animals exposed to 8901 or 19098 ppm 

were of a similar severity and incidence and of a higher severity than those seen for animals 

exposed to 4721 ppm. In consideration of the epithelial changes the olfactory epithelium was 

the most extensively affected at all three exposure levels. 

In the larynx, epithelial metaplasia, hyperplasia and keratinisation were seen in animals of all 

test article groups. As with the nasal epithelia, these changes were an adaptive response to 

produce an epithelium more resistant to irritation. Inflammation was seen in one male and 

one female exposed to 19098 ppm and one male exposed to 8901 ppm and was considered a 

consequence of irritancy. 

In general, the findings in the larynx were of a similar incidence and degree in all groups 

exposed to 2-bromo-3,3,3-trifluoropropene. 

215
Huntingdon Life Sciences

WAG0013

 Issue No: 2 

Page 10 of 10 

In the trachea, epithelial degeneration/regeneration was seen in male and female animals 

exposed to 19098 ppm and one female exposed to 8901 ppm. As in the nasal cavity, the 

degeneration was due to the irritation of the epithelium and the regeneration was part of the 

tissue repair process. Inflammatory cell infiltrate in the lamina propria was seen at a higher 

incidence and/or degree in animals exposed to 19098 ppm, compared with control animals, 

which again was considered a consequence of irritancy. 

Overall, the most marked changes were in the nasal cavity, with less severe (minimal or 

slight) changes in the larynx and trachea. For the nasal cavity, in general the changes seen in 

animals exposed to 8901 or 19098 ppm were of a similar severity and incidence. As rodents 

are obligate nose-breathers, this distribution of findings is considered accounted for by the 

nasal cavity being the first part of the respiratory tract to have contact with the test article. In 

addition, the nasal epithelium, and especially the olfactory epithelium which is anatomically 

confined to the dorsal half of the nasal cavity, is a site of metabolic enzymes and so 

metabolism of the test substance may have contributed to the severity of changes seen. 

At necropsy, a reduction in the size of the thymus and depressions or roughening of the 

stomach were noted in animals exposed to 19098 ppm. These tissues were not required to be 

examined microscopically. 

4. Conclusion 

Changes related to treatment with 2-bromo-3,3,3-trifluoropropene, were seen in the nasal 

turbinates, larynx and trachea following inhalation administration for 2 weeks. 

216
Huntingdon Life Sciences

WAG0013

217

Annex 3 Certificate of Analysis
Huntingdon Life Sciences

WAG0013

218
Huntingdon Life Sciences

WAG0013

219

Annex 4 GLP compliance statements
Huntingdon Life Sciences

WAG0013

220

EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2011
Huntingdon Life Sciences

WAG0013

221

HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2011