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Working for a better future

REPORT 

HFO-1234yf: A 28-Day Inhalation (Whole-Body Exposures) 

Toxicity Study in Minipigs 

HLS study number: 13-6434 

Sponsor reference number: 

Version ID: Final 

Issue date: 20 January 2014 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

2 

Table of Contents 

Statement of Compliance..............................................................................................................6

Signature Page .............................................................................................................................7

Quality Assurance Statement .......................................................................................................8

Details of Sponsor, Test Facility and Test Sites ...........................................................................9

Contributing Scientists ................................................................................................................10

Summary.....................................................................................................................................11

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

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

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

1.2.1 Test guidelines ...........................................................................................12

1.2.2 Good laboratory practices ..........................................................................12

1.2.3 Animal welfare act compliance ...................................................................12

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

1.4 Route, duration and frequency of administration....................................................13

1.5 Exposure level........................................................................................................13

1.6 Number of animals .................................................................................................13

1.7 Use of restraint device............................................................................................14

2. Experimental Procedure.....................................................................................................15

2.1 Study schedule and structure.................................................................................15

2.1.1 Study dates.................................................................................................15

2.1.2 Experimental outline ...................................................................................16

2.2 Test substance.......................................................................................................16

2.2.1 Test substance ...........................................................................................16

2.3 Test substance administration................................................................................17

2.4 Animal management ..............................................................................................18

2.4.1 Animals, allocation and identification..........................................................18

2.4.2 Husbandry ..................................................................................................19

2.5 Experimental evaluations .......................................................................................21

2.5.1 Daily observations (in-pen).........................................................................21

2.5.2 Dosing observations (in-chamber)..............................................................21

2.5.3 Signs...........................................................................................................21

2.5.4 Body weight ................................................................................................22

2.5.5 Food consumption ......................................................................................22

2.5.6 Clinical pathology .......................................................................................22

2.6 Postmortem ............................................................................................................24

2.6.1 Method of euthanasia .................................................................................24

2.6.2 Scheduled necropsy ...................................................................................24

2.6.3 Macroscopic examinations .........................................................................24

2.6.4 Organ weights ............................................................................................24

2.6.5 Tissues preserved and examined histopathologically ................................24

2.6.6 Preservatives..............................................................................................25

2.6.7 Processing..................................................................................................26

2.7 Computer systems .................................................................................................27

2.8 Data treatment........................................................................................................27
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

3 

2.8.1 Serial observations .....................................................................................28

2.8.2 Terminal procedures...................................................................................29

2.8.3 Statistical analysis ......................................................................................29

2.9 Data storage...........................................................................................................31

2.10 Protocol deviations .................................................................................................31

3. Results and Discussion......................................................................................................33

3.1 Exposure results.....................................................................................................33

3.2 Mortality..................................................................................................................33

3.3 Clinical observations ..............................................................................................33

3.4 Body weights ..........................................................................................................33

3.5 Food consumption..................................................................................................34

3.6 Clinical pathology ...................................................................................................34

3.6.1 Hematology ................................................................................................34

3.6.2 Clinical chemistry........................................................................................34

3.7 Organ weights ........................................................................................................34

3.8 Pathology ...............................................................................................................35

3.8.1 Mortality and macroscopic findings ............................................................35

3.8.2 Microscopic evaluation ...............................................................................35

4. Conclusion .........................................................................................................................37

5. References.........................................................................................................................38
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

4 

List of Tables 

Table 1 Daily observations - group distribution of observations.......................................40

Table 2 Dose observations - group distribution of observations.....................................102

Table 3 Clinical signs - group distribution of observations .............................................124

Table 4 Body weights - group mean values ...................................................................127

Table 5 Body weight change – group mean values........................................................129

Table 6 Visual food consumption – group distribution of observations ..........................131

Table 7 Hematology – group mean values.....................................................................133

Table 8 Clinical chemistry – group mean values............................................................149

Table 9 Organ weights – group mean absolute values ..................................................165

Table 10 Organ weights – group mean values relative (%) to body weight .....................167

Table 11 Organ weights – group mean values relative (%) to brain weight .....................169

Table 12 Macropathology – group distribution of findings................................................171

List of Appendices 

Appendix 1 Daily observations – individual observations.....................................................173

Appendix 2 Dose observations – individual observations ....................................................184

Appendix 3 Clinical signs – individual observations .............................................................190

Appendix 4 Body weights – individual values.......................................................................198

Appendix 5 Body weight change - individual values ............................................................202

Appendix 6 Visual food consumption - individual observations............................................206

Appendix 7 Hematology - individual values..........................................................................220

Appendix 8 Clinical chemistry - individual values.................................................................244

Appendix 9 Organ weights - individual values......................................................................268

Appendix 10 Macropathology – individual findings.................................................................276
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

5 

List of Annexes 

Annex 1 Certificates of analysis ......................................................................................284

Annex 2 Aerosol technology report .................................................................................295

Annex 3 Clinical pathology report....................................................................................314

Annex 4 Anatomic pathology report – organ weights ......................................................316

Annex 5 Anatomic pathology report – macroscopic and histopathology .........................318



Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

9 

Details of Sponsor, Test Facility and Test Sites 

Sponsor 

Test facility Huntingdon Life Sciences 

PO Box 2360 

Mettlers Road 

East Millstone, New Jersey 08875-2360 

Test site Ani Lytics, Inc. 

[Serum biomarker] 200 Girard Street Suite 200 

Gaithersburg, Maryland 20877-3490 

Test site University of Würzburg 

[Urine metabolite Versbacher Straße 9 

analysis] 97078 Würzburg 

Germany 

Test site HSRL Inc. 

[Anatomic pathology] 5930 Main St. 

Mount Jackson, VA 22842 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

10 

Contributing Scientists 

Senior Vice President, Safety 

Assessment 

Vice President, Bioanalytical and 

Translational Sciences 

Vice President, Toxicology Operations 

Director, Pathology 

Director, Quality Assurance 

Study Director 

Veterinarian 

Inhalation Toxicology 

Large Animal Toxicology 

Pharmacy 

Clinical Pathology and Sample Logistics 

Necropsy, Fetal Pathology and Histology

Principal Investigator, Aerosol 

Technologist 

Principal Investigator, Clinical 

Pathologist 

Principal Investigator, Organ Weight 

Evaluations 

Principal Investigator, Microscopic 

Pathologist HSRL Inc. 

Mount Jackson, Virginia 

Principal Investigator, Serum Biomarker 

Analysis Ani Lytics, Inc. 

Gaithersburg, Maryland 

Principal Investigator, Urine Metabolite 

Analysis University of Würzburg, Germany 

Peer Review Pathologist 

Experimental Pathology Laboratories 

Sterling, Virginia 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

11 

Summary 

HFO-1234yf: A 28-Day Inhalation (Whole-Body Exposures) Toxicity Study in 

Minipigs 

Göttingen minipigs (8/sex/group) were exposed by whole-body inhalation to HFO-1234yf at 

target concentrations of 0, 5,000 or 10,000 ppm (daily for 6 hours per day) for up to 28 days. 

Animal No. 2148 was euthanized before the initiation of exposures on Day 1, and was replaced 

by Animal No. 2153, who was exposed for 27 days. At the end of the treatment period, all 

surviving animals were euthanized and necropsied. Parameters evaluated during the study were: 

viability, clinical observations, body weights, food consumption, or clinical pathology (pretest 

and Days 21 and 28), organ weights, macroscopic observations and microscopic pathology. 

Mean target and achieved exposure results were as follows: 

Group Treatment Target Exposure Levels Actual Exposure Levels 

 (ppm) (mg/L) (ppm) 

 

1 Air 0 0 0 

2 HFO-1234yf 5,000 26 5145 

3 HFO-1234yf 10,000 50 10200 

 

No HFO-1234yf gas was detected in Group 1 samples. Mean atmosphere concentrations were 

3.8% and 2.0% above target for Group 2 males and females, respectively, and 2.0% above target 

for Group 3. 

At Day 29, there were no HFO-1234yf-related microscopic findings involving the heart (left 

ventricle, right ventricle and septum) and skeletal muscle (rectus femoris, psoas and soleus 

muscles as well as the diaphragm). Therefore, there was no microscopic evidence of 

cardiotoxicity or skeletal muscle toxicity following whole-body inhalation of the test substance 

at up to 10,000 ppm for 28 days in male and female Gottingen minipigs. The results of this 

study demonstrate that minipigs respond very differently to HFO-1234yf inhalation exposure as 

compared to rabbits. 

Under the condition of this study, the No-Observed-Adverse-Effect-Level (NOAEL) for 

inhalation toxicity was determined to be the actual achieved exposure level of 10,200 ppm. 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

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1. Introduction 

1.1 Objective 

The purpose of this study was to assess the cardiotoxicity and skeletal muscle toxicity of 

HFO-1234yf when administered via whole-body inhalation to minipigs for 28 days. 

1.2 Regulatory compliance 

1.2.1 Test guidelines 

This study is designed in accordance with OECD Guidelines for Testing of Chemicals No. 412 

"Subacute Inhalation Toxicity: 28 Days", Adopted 7 September 2009 (except this study was 

conducted in minipigs and only for selected parameters). 

1.2.2 Good laboratory practices 

This study was conducted in compliance with the following Good Laboratory Practices: 

Organization for Economic Cooperation and Development (OECD) Principles of Good 

Laboratory Practices ENV/MC/CHEM/(98)17 with the following exception: 

The urine metabolites analysis will not be conducted in compliance with the referenced GLPs 

because the Sponsor-subcontracted laboratory is not GLP-compliant. 

1.2.3 Animal welfare act compliance 

This study complied with all appropriate parts of the Animal Welfare Act Regulations: 9 CFR 

Parts 1 and 2 Final Rules, Federal Register, Volume 54, No. 168, August 31, 1989, pp. 36112-

36163 effective October 30, 1989 and 9 CFR Part 3 Animal Welfare Standards; Final Rule, 

Federal Register, Volume 56, No. 32, February 15, 1991, pp. 6426-6505 effective March 18, 

1991. 

1.3 Test system 

The minipig is accepted by regulatory authorities (including the European Chemicals Agency) as 

a non-rodent surrogate for humans in the detection of toxic effects. Cardiotoxicity and skeletal 

muscle toxicity were the principal endpoints of interest for this study and the minipig is 

considered to be a sensitive and representative model for human responses. In prior testing via 

inhalation exposures with this test substance, pregnant and non-pregnant female rabbits and male 

rabbits were found to be more sensitive to the test substance when compared to pregnant and 

non-pregnant female rats and male rats. This study was designed to determine whether there is a 

repeated exposures (4 weeks instead of the prior tested 2 weeks) sensitivity in minipigs with this 

test substance since the minipig is a more appropriate large animal model for safety evaluation 

for this test substance. 

. 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

13 

1.4 Route, duration and frequency of administration 

The inhalation route is a potential route of human exposure to this test substance. The duration 

and frequency of the exposures in this study were intended to insure adequate repeat-exposure 

toxicity characterization for the test substance in minipigs, as compared to prior data in rabbits. 

In prior rabbit testing (HLS study 11-6387), cardiotoxicity and skeletal muscle toxicity were seen 

after 1-2 weeks of exposures. Similar effects were not observed in rabbits following acute 

exposure, indicating that repeated exposure was needed to produce these effects. However, in a 

pilot minipig study (HLS study 13-6432), cardiotoxicity and skeletal muscle toxicity were not 

seen after 2 weeks of exposure. This study confirmed that minipigs responded similar to rats as 

compared to rabbits. 

1.5 Exposure level 

A prior 28-days inhalation study (HLS 11-6387) in male and non-pregnant female rabbits resulted 

in unscheduled deaths/sacrifices at 4500 ppm (1 female) and 5500 ppm (1 male, 1 female). 

Clinical signs prior to euthanasia included decreased activity, ano-genital staining decreased 

fecal volume and unformed stool. Exposure to HFO-1234yf was associated with microscopic 

findings in the heart and skeletal muscle at ≥1000 ppm with males more frequently affected than 

females. The NOEL (No-Observed-Effect-Level) based on anatomic pathology findings was 

500 ppm in males and 1000 ppm in females. 

No mortality or toxicity was seen in male and female minipigs (HLS 13-6432) from up to 

2 weeks of exposures up to a 10,000 ppm level of the test substance. 

Based on the data in rabbit and minipig toxicity studies discussed previously, a high exposure 

level of 10,000 ppm was selected for this study. Since several animals did not have a cause of 

death determined, it is not certain that the lethality was due to the cardiac lesions. As such, there 

was a need to evaluate the different endpoint(s) using this high concentration (for example – the 

effects may have been due to a saturation of a metabolic pathway). If only lower exposure levels 

were used, this (or a similar) finding might have been missed. A lower exposure level of 

5,000 ppm was also selected to provide a range of possible test substance effects. The potential 

effects in minipigs following inhalation of 5,000 and 10,000 ppm HFO-1234yf for following 

28 days had not been previously investigated. 

1.6 Number of animals 

The number of animals in this study was considered the minimum necessary for statistical and 

scientific reasons. The purpose of this study was to monitor for toxicity of the test substance and 

to compare the data to the 28-day rabbit inhalation toxicity study 11-6387 to help determine the 

appropriate animal model for use in human risk assessment. The number of animals/sex/group 

(8) for this 28 days toxicity evaluation was considered the minimum number that would account 

for the following: 

(1) The expected variability among animals as noted during the 14-day preliminary study 

13-6432 in minipigs. 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

14 

(2) To provide an adequately robust data set considering the expected subtlety of the possible 

cardiotoxic and skeletal muscle effects as noted in the 28-day rabbit study 11-6387. In 

the 14-day preliminary study 13-6432 in minipigs, elevations in selected biomarkers and 

histological effects were observed in some animals in all exposure groups, including 

pretest and controls. The exposure duration for this study was twice as long and 

sufficient animals were needed to discriminate test substance-related effects, if any, from 

background variations. Due to the potential for subtle effects, it was important in this 

definitive study that sufficient animals were evaluated to produce robust results and 

conclusions. 

(3) The historical control database for minipigs exposed via whole-body inhalation exposure 

is very limited (only one study: the 14-day preliminary minipig inhalation study 13-6432) 

at this laboratory. The proposed group sizes were thus necessary to better define the 

normal range of variability for the parameters evaluated under this exposure study in this 

species. 

(4) A robust study design such as proposed for this study provided an enhanced degree of 

confidence in the results to best protect the potentially exposed human population. 

Two test substance-treated groups receiving low and high exposure levels and a negative air 

control group were considered the minimum number of groups necessary to provide a range of 

effects and allow for extrapolation of results to humans. 

1.7 Use of restraint device 

Animals were restrained by a technician in a V-trough for the clinical pathology bleeding since 

bleeding the animals without this restraint was impractical. Formal habituation to the v-trough 

for purposes of blood collection was not necessary because the animals were handled frequently 

before they were bled. 

Animals were restrained in the whole-body inhalation exposure chambers (each chamber was 

considered a restraint device because of its limited floor space of ~10 square-feet for 4 pigs) 

during the daily exposures since exposing the animals without this restraint was impractical (for 

example to expose the animals in their pens would require impractical quantities of test 

substance in the room air and would create a health hazard for the technical staff). Also, the 

exposures method simulated the exposures method for the prior rabbit and minipig inhalation 

studies. Animals were habituated to the inhalation exposure chambers for purposes of 

minimizing stress during exposures (to include ~1 hour on the 1st day, ~1 hour on the 2nd day 

preceding the 2nd day of pretest blood collection (bleeding done within 1 hour after removal from 

the chamber and before the pm feeding) in preparation for this study. 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

15 

2. Experimental Procedure 

2.1 Study schedule and structure 

2.1.1 Study dates 

Study initiation: 6 September 2013 (Date Study Director signed the Protocol) 

 

Experimental start date: 17 September 2013 (Date of first data collection) 

 

Animal receipt: 17 September 2013 

 

Dosing initiation: 

 males 2 October 2013 

 females 3 October 2013 

 

Dosing termination: 

 males 29 October 2013 

 females 30 October 2013 

 

Necropsy: 

 males 30 October 2013 

 females 31 October 2013 

 

Experimental completion date: 17 January 2014 (Date of last data collection) 

 

Study completion: 20 January 2014 (Date Study Director signed the Final 

Report) 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

16 

2.1.2 Experimental outline 

The test substance was administered via whole-body inhalation to minipigs for up to 

28 consecutive days. Animal No. 2148 was euthanized before the initiation of exposures on 

Day 1, this animal was replaced by Animal No. 2153 who was exposed for only 27 days. 

The study consisted of 1 control and 2 treated groups identified as follows: 

Group Treatment Target Exposure Levels Number of animals 

(ppm) (mg/L) Male Female 

 

1 Control 0 0 8 8 

2b HFO-1234yf 5,000 26 8 8 

3 HFO-1234yf 10,000 50 8 8 

 a

Exposures were 6 hours per day for 7 days per week for up to 28 days. Exposure levels are expressed as 

ppm (part per million of test substance in air) and equivalent mg/L (mg of test substance per Liter of air) of 

test substance. Exposure results are reported as in terms of PPM. Animals in Group 1 were similarly 

exposed to air only. 

2.2 Test substance 

2.2.1 Test substance 

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

records. Certificates of Analysis for each lot number of test substance are presented in Annex 1.

The following information is given in summary: 

Test substance: HFO-1234yf (2,3,3,3 – Tetrafluoropropene) 

MW: 114 grams/mole 

CAS No.: 754-12-1 

Test Substance Category: Industrial chemical used as a possible refrigerant 

Manufacturer/Supplier: 

Description: Liquid (compressed gas) 

Storage conditions: Room temperature 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

17 

Lot numbers: 050714C 

051181C 

053912C 

058794C 

058876C 

060571C 

060648C 

060668C 

061520C 

062669C 

Purity: 99.99% 

Date received: 13 September 2013 

Expiration date: 26 August 2014 

Documentation of the identity, strength, purity, composition, stability, and method of synthesis, 

fabrication, and/or derivation of each batch of the test substance and the maintenance of these 

records were the responsibility of the Sponsor. 

A sample from each lot of test substance is stored in the Archives of the Testing Facility under 

conditions specified for test substance storage. 

The unused portion of the test substance and any empty test substance containers will be returned 

to the Sponsor following issue of the final report of the final study with this test substance. 

The test substance was used as supplied. 

Detailed records of test substance usage were maintained in the study raw data. 

2.3 Test substance administration 

Route Inhalation – whole-body. 

Controls (Group 1) Air only. 

Training for dosing (wholebody) The animals were acclimated to the method of restraint for two 

1 hour sessions during the pretest period. 

Duration of daily exposure 6 hours. 

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

are presented in Annex 2. 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

18 

2.4 Animal management 

2.4.1 Animals, allocation and identification 

Animals 

Strain/Species Göttingen Minipig (juvenile) 

Supplier Marshall BioResources 

North Rose, New York 14516 

Number of animals Received Placed on test 

 Total: 52 48 

 Males: 26 24 

 Females: 26 24 

Pretest period The pretest period was approximately 2 weeks. Pretest 

procedures were not performed until animals had been allowed 

to stabilize for approximately 1 week. Prior to exposure 

initiation, all animals were examined by the staff veterinarian to 

ascertain suitability for study. The staff veterinarian examined 

the general health of each animal and determined the need for 

medical treatment or prophylaxis. 

Age of at start of treatment Approximately 3 to 4 months 

Weight range at the start of Males: 6.0 kg to 8.9 kg 

treatment 

Females: 6.3 kg to 9.7 kg 

Veterinary procedures 

Veterinary care Animals were monitored by the technical staff for any conditions 

requiring possible veterinary care and treated as necessary. 

Miscellaneous, non-test substance-related veterinarian 

evaluations and treatments (as necessary) for individual animals 

were reviewed by the study director and are documented in the 

study file. 

Allocation and identification 

Allocation More animals than required for the study were purchased and 

stabilized. Animals considered suitable for study were allocated 

to the control, treated or spare animal groups upon arrival. 

Animal No. 2148 was euthanized before the initiation of 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

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exposures on Day 1 and this animal was replaced by Animal 

No. 2153. Information as to the disposition of all animals not 

utilized in the study is maintained in the study file. 

Identification of animals Each animal bore an ear tag identification applied by the 

Supplier. Following allocation to the study, the assigned number 

was applied (marked on a hind limb and the head with 

permanent marker pen). The animal number plus the study 

number comprised the unique animal number for each animal. 

Each animal’s pen was provided with a card, color-coded for 

exposure level identification and bearing the study number and 

animal number information. 

2.4.2 Husbandry 

Facilities management/animal husbandry 

Currently acceptable practices of good animal husbandry were followed e.g., Guide for the Care 

and Use of Laboratory Animals; National Academies Press, 2011. Huntingdon Life Sciences, 

East Millstone, New Jersey is fully accredited by the Association for Assessment and 

Accreditation of Laboratory Animal Care International (AAALAC). 

Environmental control 

Light/dark cycle A twelve hour light/dark cycle was provided and controlled via 

an automatic timer. 

Temperature and relative 

humidity 

The temperature was monitored and maintained within the range 

of 22 to 27 °C. The relative humidity was monitored and 

maintained within the range of 30 to 70%. 

Although conditions were occasionally outside the indicated 

ranges, these deviations were minor and/or of short duration and 

were not considered to have influenced the health of the animals 

and/or the outcome of the study. 

Air changes Ten to fifteen (10 to 15) fresh air changes per hour are 

considered the acceptable standard. Air changes were 

periodically monitored by plant maintenance and the records 

were reviewed by the veterinarian. 

Animal housing and environmental enrichment 

Housing during nonexposure periods Pens by sex and exposure group. 

Number of animals per pen Up to 4 of the same sex per pen. The number of animals in a pen 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

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was reduced due to mortality. 

Housing during exposure 

periods 

Group housed in chamber (~10 square-feet for up to 4 pigs). 

Animals were single-housed in the chamber due to injury and 

resumed group-housing following approval by the veterinarian as 

detailed below. 

Beginning on Day 15, Animal No. 1148 was singled-housed in 

the chamber due to injury to its penis. 

On Day 13, Animal No. 1145 was singled-housed in the chamber 

due to injury to its tail. 

For the entire study, Animal No. 2153 was singled-housed in the 

chamber because this animal was not previously housed with 

other animals in the group. 

Environmental enrichment Animals were exercised in accordance with the Animal Welfare 

Standards; Final Rule (9 CFR Part 3), following the Testing 

Facility’s Standard Operating Procedures. 

Feed 

Diet Certified Miniswine Diet, No. 7037 

Harlan Teklad, Madison, Wisconsin 

Diet during exposure 

periods 

None. 

Availability Minipigs were fed 2 times daily with the approximate total 

amount of 250 grams per day. Feed was presented at 

approximately the same times each day including approximately 

one-half of the feed before exposures and approximately one-half 

of the feed after exposures on the days of exposures. On 

Days 21 and 28 when clinical pathology sample collections were 

scheduled after the exposures, the animals were fed 

approximately one-half of the feed before exposures and 

approximately one-half of the feed after the blood and urine 

collections. 

On occasion, diets were supplemented with pieces of fruit and 

with “treats” (such as small amounts of fruit juice, or other 

acceptable food). 

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

by the manufacturer. Results were provided to the Testing 

Facility and are maintained on file at the Testing Facility. There 

were no known contaminants in the feed that were expected to 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

21 

interfere with the results of this study. 

Water 

Supply Potable water from the public supply (New Jersey-American 

Water Company, Cherry Hill, New Jersey) via an automated 

watering system. 

Availability Without restriction (except during the periods of urine 

collection). 

Water during exposure 

periods 

None. 

Analysis Water analyses are conducted by New Jersey-American Water 

Company, Cherry Hill, New Jersey (Raritan-Millstone Plant) to 

ensure that water meets standards specified under the EPA 

Federal Safe Drinking Water Act Regulations (40 CFR Part 141). 

In addition, water samples are collected biannually from 

representative rooms in the Testing Facility; chemical and 

microbiological water analyses are conducted on these samples 

by a subcontract laboratory. Results of all water analyses are 

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

contaminants in the water which were expected to interfere with 

the results of this study. 

2.5 Experimental evaluations 

2.5.1 Daily observations (in-pen) 

Animals were observed in their pens twice daily for mortality and general condition. During the 

treatment period animals were observed twice daily for signs of toxic or pharmacologic effects. 

Animals in extremely poor health or in a possible moribund condition were identified for further 

monitoring and possible euthanasia. 

2.5.2 Dosing observations (in-chamber) 

Animals were monitored continuously during each exposure period and observations 

individually recorded at least twice, approximately 2 hours and 4 hours into the exposure. 

2.5.3 Signs 

Animals were removed from their pens and examined twice pretest and weekly during the study 

period. Examinations included observations of general condition, skin and fur, eyes, nose, oral 

cavity, abdomen and external genitalia as well as evaluations of respiration. 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

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2.5.4 Body weight 

Animals were removed from their pens and weighed 3 times pretest, weekly during treatment 

and terminally (after fasting). Terminal, fasted body weights were obtained just prior to 

necropsy. 

2.5.5 Food consumption 

A visual estimate of the amount of food consumed per day was made for each minipig (after the 

PM feeding interval) starting during the 2nd week after animal receipt and continuing throughout 

the treatment period, except when animals are fasted for terminal sacrifice. 

2.5.6 Clinical pathology 

Blood obtained via the vena cava from unanesthetized animals was used to analyze hematology, 

and clinical chemistry parameters for up to 8 animals/sex/group twice pretest and after exposures 

on Days 21 and 28 (within 1 hour after removal from the exposure chamber and before PM 

feeding). Animals were fasted for at least 7 hours (including during the 6 hours of exposure on 

exposure days) prior to each scheduled blood collection interval. The animals were held or 

placed into a v-trough for each sampling. The behavior of all animals during blood collection 

was recorded in the raw data including any unusual or excessive struggling. 

Hematology 

Blood samples (approximately 2 mL) were collected into tubes containing K2EDTA 

anticoagulant and analyzed for the following using a Siemens ADVIA 120 Hematology 

Analyzer: 

Hemoglobin concentration (HGB) 

Hematocrit (HCT) 

Erythrocyte count (RBC) 

Platelet count (PLT) 

Mean corpuscular volume (MCV) 

Mean corpuscular hemoglobin (MCH) 

Mean corpuscular hemoglobin concentration (MCHC) 

Red cell distribution width (RDW) 

Total leukocyte count (WBC) 

Reticulocyte count (RETIC) 

Differential leukocyte count 

Neutrophils (ANEU) 

Lymphocytes (ALYM) 

Eosinophils (AEOS) 

Basophils (ABASO) 

Monocytes (AMONO) 

Large unstained cells (ALUC) 
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

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A peripheral blood smear was prepared for each animal at each blood collection interval and was 

available for confirmation of automated results and/or other evaluations deemed necessary by the 

Clinical Pathologist. 

Clinical chemistry 

Blood samples (approximately 3 mL) were collected into tubes with no anticoagulant, allowed to 

clot and centrifuged to obtain serum then split into 2 aliquots. An approximately 400 μL aliquot 

of serum was designated for clinical chemistry analysis by the Testing Facility. An 

approximately 800 μL aliquot of serum was designated for cardiac biomarker analysis, stored 

frozen at – 80 °C ± 10 °C and shipped on dry ice to Ani Lytics, Inc., Gaithersburg, Maryland for 

analysis. 

Blood samples (approximately 2 mL) were collected into tubes containing K3EDTA 

anticoagulant and centrifuged to obtain plasma. One aliquot of plasma (approximately 800 μL) 

was stored frozen at – 80 °C ± 10 °C for possible future evaluations after the issue of the final 

report. 

Clinical chemistry blood samples were collected from Animal No. 2147 prior to sacrifice for 

welfare reasons. 

Tests performed by the Testing Facility 

Samples were analyzed for the following using a Siemens ADVIA 1800 Chemistry Analyzer: 

Aspartate aminotransferase (AST) (Kinetic - Modified Bergmeyer) 

Alanine aminotransferase (ALT) (Kinetic - Modified Bergmeyer) 

Creatine kinase-Total (CK) (UV, NAC-activated Kinetic) 

Tests performed by Ani Lytics, Inc.:

Serum cardiac biomarker samples were frozen and stored at – 80 °C ± 10 °C until shipped on dry 

ice. 

Creatine kinase-Total (CK) (UV, NAC-activated Kinetic) 

creatine kinase-MB (CK(MB)) (electrophoretic isoenzyme separation) 

creatine kinase-MM (CK(MM)) (electrophoretic isoenzyme separation) 

myoglobin (MYOGLOBI) (ELISA) 

cardiac troponin I (TPI) (ELISA) 

Serum samples for possible future evaluations were stored frozen at – 80 °C ± 10 °C until the 

Sponsor determines the final disposition after the issue of the final report. 

Urinalysis 

Urine obtained via an up to 15-hour overnight collection period will be analyzed for up to 8 

animals/sex/group once pretest and after exposures on Days 21 and 28. Animals were 

transferred to cages, fasted and water-deprived during the collection periods. 
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Urine was collected into ice-chilled containers overnight (approximately 13 to 15 hours) from 

urine pans beneath each individually-housed animal’s cage. Urine samples were processed (total 

volume measured), split into 2 samples (primary and backup, approximately 10 mL each) and 

frozen in dry ice/alcohol and stored at –20 °C ± 10 °C. The primary samples were shipped on 

dry ice to Paragon Global Services Ltd., Alconbury Cambridgeshire, UK for further shipment to 

University of Würzburg, Germany for urine metabolites analysis. The urine metabolites data 

will be communicated to the Study Director and maintained in the study data but not presented in 

the report. The backup samples will remain frozen until the Sponsor determines the final 

disposition after the issue of the final report. 

2.6 Postmortem 

2.6.1 Method of euthanasia 

Animals sacrificed for welfare reasons or at the scheduled sacrifice interval were sedated with 

telazol and xylazine followed by intravenous overdose of administration of sodium pentobarbital 

and then exsanguination. 

2.6.2 Scheduled necropsy 

Necropsy was performed on up to 8 animals/sex/group after animals had been treated for up to 

28 days. Animals were fasted overnight prior to necropsy. Necropsy schedules were established 

in order to assure that approximately equal numbers of males and females from each group were 

examined at similar times of the day throughout the necropsy period. A pathologist was present 

during the scheduled necropsy examinations. 

2.6.3 Macroscopic examinations 

Complete macroscopic examinations were performed on all animals including the unscheduled 

decedent. The macroscopic examination included examination of the external surface and all 

orifices; the external surfaces of the brain and spinal cord; the organs and tissues of the cranial, 

thoracic, abdominal and pelvic cavities and neck; and the remainder of the carcass for the 

presence of macroscopic morphologic abnormalities. 

2.6.4 Organ weights 

Organs indicated in Text Table 2.6.5-1 were weighed for all animals at the scheduled sacrifice 

interval. Prior to weighing, the organs were carefully dissected and properly trimmed to remove 

adipose and other contiguous tissues in a uniform manner. Organs were weighed as soon as 

possible after dissection in order to avoid drying. Paired organs were weighed together. 

2.6.5 Tissues preserved and examined histopathologically 

The tissues listed in Text Table 2.6.5-1 were obtained from all animals and preserved. In 

addition, slides of the indicated tissues were prepared and shipped to the pathologist and 

examined microscopically. Any abnormalities not noted during macroscopic examinations 

which were seen during histology processing were recorded. 
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Text Table 2.6.5-1: Tissues weighed, preserved and examined microscopically 

ORGAN NAME WEIGHED PRESERVED 

EXAMINED 

MICROSCOPICALLY 

brain (medulla, pons, cerebrum and cerebellum) X X 

heart (left and right ventricles and septum) X X X 

kidneys X X 

liver X X 

lungs (with mainstem bronchi) X X 

skeletal muscle (diaphragm and bilateral

psoas and bilateral rectus femoris and soleus) 

 

X X 

tissues with macroscopic findings including 

 tissue masses 

 

X a

Qualitative examination (no differential count). 

2.6.6 Preservatives 

All tissues were preserved in 10% neutral buffered formalin (NBF). Lungs were infused with 

10% NBF prior to their immersion into a larger volume of the same fixative. 

The rectus femoris was removed in its entirety from each hind leg, laid flat on a stiff card and 

stapled at either end of the card to maintain its position during fixation. A thin longitudinal slice 

of rectus femoris was removed and flash frozen for possible future analysis. 

Each psoas muscle was removed in its entirety and stapled to a stiff card (no psoas muscle was 

frozen). 

The calf muscle that gives rise to the calcaneal tendon (including both heads of the 

gastrocnemius muscle and the underlying soleus muscle) was removed from both hind legs. 

Each muscle mass was collected by cutting the common calcaneal tendon, reflecting it toward 

the femorotibial joint and cutting the muscles at their origin on the distal femur (gastrocnemius) 

and head of the fibula (soleus). The soleus was not separated from the gastrocnemius. The 

muscles were stapled to a stiff card (no soleus muscle was frozen). 

The diaphragm was sampled on the left side of the body from the thoracic wall to the esophageal 

hiatus and stapled to a card to maintain its position during immersion fixation (no diaphragm was 

frozen). 

Tissues for possible future analysis (hearts and rectus femoris) were archived. The Sponsor will 

determine the disposition of these materials (i.e. archival, shipment or disposal of samples). 

To best remove and preserve the hearts to reduce as much as possible any artifacts in the tissues: 

1. The animal was sedated in the study room and euthanized in the necropsy lab. 

2. The heart was quickly removed from the animal, weighed and the 4 chambers were 

flushed with chilled saline using a perfusion pump in order to cool the heart rapidly. 
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3. When the hearts stop beating, the left and right ventricles were finely dissected for best 

fixation. A longitudinal incision was made parallel to the two main bundles of papillary 

muscles in the left ventricle and the largest papillary muscle bundle in the right ventricle. 

The purpose of these incisions was to allow more rapid fixation of the papillary muscles. 

The incisions were made adjacent to the muscle bundles to facilitate formalin penetration 

but not so close that they interfere with accurate trimming. 

4. A thin longitudinal slice of left ventricular free wall was made from both margins of the 

opening incision in the left ventricle. The slice from the left margin was flash frozen with 

liquid nitrogen and stored for possible future analysis. 

5. The slice from the right margin was sampled for electron microscopy and preserved in 

modified Karnovsky’s fixative for possible future analysis. Care was taken to avoid 

damaging the papillary muscles near the opening incisions since these areas were sampled 

for histopathology. 

6. After removing tissue for freezing and electron microscopy, the hearts were placed into 

chilled formalin for the duration of the necropsy and transferred to the bucket of formalin 

at the conclusion of the necropsy. 

2.6.7 Processing 

Tissues (heart and muscle) were processed, sectioned at approximately 5 microns, and examined 

microscopically for all animals. 

A total of eight (8) longitudinal sections were sampled from the heart, 2 from the right 

ventricular free wall, 2 from the interventricular septum and 4 from the left ventricular free wall. 

The exact position of these sections was indicated in a trimming guide in the study file. Sections 

were processed in large cassettes with the top surface of the tissue sample marked with surgical 

ink to ensure proper positioning at embedding. 

The rectus femoris was sampled at the midbody and towards either end (but away from the 

stapled areas) for a total of 3 sample areas. At each area, both a cross section and a longitudinal 

section of the muscle tissue were taken. A cross section and a longitudinal section were made in 

the middle of the psoas muscle (1 sample area). A cross section and a longitudinal section were 

made in the middle of the soleus muscle (1 sample area). A cross section and a longitudinal 

section were made from a representative area of the diaphragm. 

Each block was serial sectioned to prepare 2 duplicate sets of slides. 

One set of the duplicate slides for the hearts and muscles were shipped to the pathologist at 

HSRL. The slides were then transferred to the Peer Review Pathologist. The histopathology 

report including the peer review statement is presented in Annex 5. 

The other set of the duplicate slides for hearts were shipped to 

 for an informal peer review (no 

report was required). 

All slides were returned to the Testing Facility upon completion of examination. 
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2.7 Computer systems 

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

data include: 

System Version Use 

AmegaView Environmental 

Monitoring System 

3 Monitoring of environmental conditions within the 

pharmacy 

ClinAxys II 1.1.5.1.2 Acquisition, processing and reporting of clinical 

pathology data from a variety of instruments 

Liberate Reporting System 2.2 Generation of report tables in MS Word formats for 

data collected using the Xybion Path/Tox System 

and ClinAxys II 

Liberate Reporting System - 

Global 

1 Generation of report tables in MS Word formats for 

data collected using the Xybion Pristima® System 

Microsoft Word and/or 

Excel 

 Additional formatting of report tables. 

Quasar 1.4 Statistical analysis of data 

REES Scientific 

Environmental Monitoring 

System 

WIN 1.5 Monitor and record temperature, humidity and/or 

light intensity readings of applicable rooms as well 

as the temperature readings of applicable 

refrigerators and/or freezers 

Xybion Pristima® System 6.2 Documentation of receipt and tracking of substances 

received by Pharmacy; direct online capture of inlife and postmortem evaluations 

PathData® Systems 

(HSRL only) 

V6.2c2 Direct online capture of microscopic pathology data 

and for the generation of report tables for the 

microscopic pathology data. 

2.8 Data treatment 

Individual animal data presented in this report may be rounded. Unrounded individual animal 

data were used to calculate the reported mean and standard deviation values. Therefore, use of 

the reported individual values to reproduce means, standard deviations and/or to perform any 

subsequent calculations may produce minor discrepancies between the calculated values and 

those presented in this report. 
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Group means and standard deviations for calculated data points (i.e. body weight change) were 

derived from individual animal/cage values. 

The data collection system used for collecting the in-life and post-life data divides the study into 

phases. The start of each phase begins with Day 1/Week 1. The phases presented in this report 

are: 

Pre-Treatment: Begins the day the animals arrive. Phase days/weeks are labeled 

P1, P2, P3, etc. During this phase, animals are not being dosed and 

they have not been assigned their permanent identification 

numbers. 

Treatment: Begins for each animal when it receives its first administration of 

test/control substance as specified in the protocol. 

The following abbreviations are used in this report: 

M Male(s) 

F Female(s) 

~ approximately 

Conc concentration 

Lpm Liters per minute 

mg/L milligrams of test substance per Liter of air 

ppm parts per million 

HLS Huntingdon Life Sciences 

MW molecular weight 

The death codes used have the following meanings: 

S1 Scheduled Euthanasia 1 

WE Euthanized for welfare reasons 

2.8.1 Serial observations 

Clinical observations 

Clinical observations are presented for each animal that showed signs, providing detail of the 

type of sign, day or week of occurrence and information on the duration of the sign applicable. 

AM Morning (pre-exposure) Observation 

PM Afternoon (post-exposure) Observation 

US PM Unscheduled Afternoon (post-exposure) Observation 

2 Hours 2 Hours into Exposure 

4 Hours 4 Hours into Exposure 

US1 1st Unscheduled Observation 

US2 2nd Unscheduled Observation 
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Hematology 

The following abbreviations were used: 

NS No specimen 

Sample condition (SC) was described using the following abbreviations: 

CLSE Severe platelet clumping noted 

CLSL Slight platelet clumping noted 

NCLP No clumping 

Blood chemistry 

CK values below the analytical limit (<7800) are excluded from calculations. 

The following abbreviation was used: 

NS No specimen 

Sample condition (SC) was described using the following abbreviations: 

SHI Slightly hemolyzed 

Absolute blood creatine kinase-MB (CK(MB)) was calculated as: 

creatine kinase-total x (creatine kinase-MB ÷ 100) 

Absolute blood creatine kinase-MM (CK(MM)) was calculated as: 

creatine kinase-total x (creatine kinase-MM ÷ 100) 

2.8.2 Terminal procedures 

Organ weights 

Organ weights were presented as absolute, relative to terminal body weight using the weight 

recorded on the day of necropsy and relative to brain weight. 

2.8.3 Statistical analysis 

All statistical analyses were carried out separately for males and females. For all other 

parameters, the analyses were carried out using the individual animal as the basic experimental 

unit. 
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The following data types were analyzed at each timepoint separately: 

body weight 

body weight change from interval to interval 

cumulative body weight change from baseline 

hematology 

clinical chemistry 

organ weights 

The parameters to analyze were identified as continuous, discrete or binary. Test-substance 

treated groups were then compared to the control using the following procedures. 

Continuous Parameters 

For comparisons involving more than two groups, if Bartlett's test for variance homogeneity 

(Bartlett, 1937) was not significant at the 1% level, then parametric methods were applied. If the 

F1 approximate test for monotonicity (described below) of dose-response was not significant at 

the 1% level, Williams' test (Williams’, 1971, 1972) for a monotonic trend was applied. If the 

F1 test was significant, suggesting that the dose-response was not monotone, Dunnett's test 

(Dunnett, 1955, 1964) was performed instead. If Bartlett's test was significant at the 1% level, 

then logarithmic and square-root transformations were tried. If Bartlett's test was still 

significant, then non-parametric methods were applied to mean ranks. If the H1 approximate test 

for monotonicity (described below) was not significant at the 1% level, Shirley's test for a 

monotonic trend (Shirley, 1977) was applied. If the H1 test was significant, then Steel's test 

(Steel, 1959) was performed instead. (Bartlett, 1937) 

The F1 test is designed to detect significant departure from monotonicity of an ordered set of 

means. The test statistic compares the mean square, NMS, for the deviations of the absolute or 

adjusted means from the maximum likelihood means, calculated under a constraint of 

monotonicity, with the usual error mean square, EMS. The null hypothesis is that the true means 

are monotonically ordered. The test statistic is F1 = NMS/EMS which can be compared with 

standard tables of the F-distribution with 1 and error degrees of freedom. 

The H1 test is the non-parametric equivalent of the F1 test. The test statistic compares the nonmonotonicity sums of squares, NRSS, for the deviations of the mean ranks from the maximum 

likelihood mean ranks, with the non-parametric equivalent of the error sums of squares, 

ERSS=N(N+1)/12. The test statistic is H1 = NRSS/ERSS which can be compared to standard 

tables of the χ2-distribution with 1 degree of freedom. 

Discrete Parameters 

For comparisons involving more than two groups, if the Jonckheere-Terpstra test (Jonckheere, 

1954) was significant at the 5% level, then the direction of the trend was established and onetailed step-down testing in this direction was performed. If the Jonckheere-Terpstra test was not 

significant at the 5% level, then the Kruskal-Wallis test (Kruskal and Wallis, 1952, 1953) was 

applied. If the Kruskal-Wallis test was significant at the 5% level, the test-substance treated 
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groups were compared to the control using exact Wilcoxon rank sum tests (Wilcoxon, 1945), 

otherwise, no further comparisons were made. 

Binary Parameters 

For comparisons involving more than two groups, if the Cochran-Armitage test (Armitage, 1955) 

was significant at the 5% level, then the direction of the trend was established and one-tailed 

step-down testing in this direction was performed. If the Cochran-Armitage test was not 

significant at the 5% level, then the χ2 test (Armitage, et. al., 2002) was applied. If the χ2 test 

was significant at the 5% level, the test-substance treated groups were compared to the control 

using Fisher’s Exact tests (Fisher, 1973), otherwise, no further comparisons were made. 

Statistical annotation 

Significant differences between control and treated groups were expressed at the 5% (p<0.05) or 

1% (p<0.01) level. The key to annotations used on the tables that contain statistical results is 

given below. 

Av Groups compared during the pretreatment period using Analysis of variance. 

Wi Treated groups compared with Control using Williams’ test. 

 

* p<0.05 

** p<0.01 

2.9 Data storage 

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

and the original final report will be maintained in the Archives of the Testing Facility upon 

completion of the study. All slides were returned to the Testing Facility following microscopic 

examination and peer review. The Sponsor will determine the final disposition of these 

materials. Electronic raw data generated at the Testing Facility are maintained on the computer 

system on which the data application resides or archived off-line. 

Biological samples collected for clinical pathology will be stored for up to 6 months after 

completion of assays and then discarded. 

All documentation, samples and data pertaining to the portions of the study performed by 

Principal Investigators will be maintained by the Test Sites’ according to their SOPs. 

2.10 Protocol deviations 

The following protocol deviations occurred during the study but were not considered to have 

compromised the validity or integrity of the study: 

1. During the 1st interval of pretest clinical pathology blood collection, animals were fasted 

for more than 7 hours prior to blood collection (fasted at 1002, blood collection at 

1716 to 1823 and fed at 2016). 
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2. During the 2nd interval of pretest clinical pathology blood collection, Group 1 male 

animals were not fasted for at least 7 hours prior to blood collection but were fasted for 

6 hours, 11 minutes to 6 hours, 45 minutes. 

3. On Days 2 to 5 of the pretest period, due to scheduling error, visual food consumption 

was measured and recorded although the protocol required food consumption to be 

estimated starting the 2nd week after animal receipt. The data recorded on these days was 

removed from Pristima (but is available for review in the audit trail) and is not reported. 

4. On Day 9 of the pretest period, documentation indicating the behavior of all animals 

during clinical pathology blood collection was not recorded; therefore it cannot be 

confirmed that the protocol requirement was met. 

5. For animal identification, the supplier ear tags were not removed and the Testing 

Facility’s animal number was not applied by tattoo; the Testing Facility’s animal number 

was applied by permanent marker pen. 

6. On Day 14, Animal No. 1148 was given an exposure holiday because of health concerns. 

7. On Days 16 and 17, Animal No. 1148 was exposed for 353 and 268, respectively, instead 

of the required duration of 360 minutes (6 hours). The exposures were terminated early 

for this animal because he was able to bypass the single-restraint compartment in the 

chamber and join the group-housed animals in the chamber. 

8. Beginning on Day 15, Animal No. 1148 was singled-housed in the chamber due to injury 

to its penis. 

9. On Days 18, 19 and 20, the chamber housing Animal No. 1148 had an average 

temperature of 19.1 °C to 19.6 °C; however, the protocol required a temperature range of 

20 °C to 27 °C. 

10. Due to oversight, Animal Nos. 1145, 1146, 2145 and 3145 were sedated and euthanized 

in the animal room rather then transferred to the necropsy lab for sedation and euthanasia. 

11. Documentation was not recorded for Animal Nos. 1145, 1146, 1147, 2145, 2146, 3145, 

3146 and 3147 to indicate that after removing tissue for freezing and electron microscopy, 

the hearts were placed into chilled formalin for the duration of the necropsy; therefore, it 

cannot be confirmed that the protocol requirement was met. 

12. The protocol did not require that CK-BB be analysed by the Subcontractor however this 

parameter was analysed by the Subcontractor and included with their raw data transfer to 

HLS. This data was not included in the HLS report but was reviewed by the HLS clinical 

pathologist and not considered to reflect any treatment-related effects. 
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3. Results and Discussion 

3.1 Exposure results 

(Annex 2) 

Mean target and achieved exposure results were as follows: 

Group Treatment Target Exposure Levels Actual Exposure Levels 

 (ppm) (mg/L) (ppm) 

 

1 Air 0 0 0 

2 HFO-1234yf 5,000 26 5145 

3 HFO-1234yf 10,000 50 10200 

 

No HFO-1234yf gas was detected in Group 1 samples. Mean atmosphere concentrations were 

3.8% and 2.0% above target for Group 2 males and females, respectively, and 2.0% above target 

for Group 3. 

The mean nominal concentration of 19100 ppm was 7% less than the overall combined IR 

measured concentration of 20600 ppm for Group 2. The mean nominal concentration of 

39200 ppm was 4% less than the overall combined IR measured concentration of 40800 ppm for 

Group 3. This was considered a reasonable experimental difference as seen previously at this 

laboratory. 

3.2 Mortality 

There were no test substance-related unscheduled deaths or sacrifices. 

A Group 2 male animal #2147 was euthanized on Day 16 due to a severely prolapsed rectum. 

All other animals survived until their scheduled sacrifices. 

3.3 Clinical observations 

(Table 1, Table 2, Table 3, Appendix 1, Appendix 2 and Appendix 3) 

There were no test substance-related clinical signs. 

3.4 Body weights 

(Table 4, Table 5, Appendix 4 and Appendix 5) 

When compared to control animals, there were no test substance-related effects on body weights. 
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3.5 Food consumption 

(Table 6 and Appendix 6) 

When compared to control animals, there were no test substance-related effects on food 

consumption. 

3.6 Clinical pathology 

3.6.1 Hematology 

(Table 7, Appendix 7 and Annex 3) 

There were no test substance-related effects. 

All differences from controls or Pretest were not considered to be test substance-related because 

they were small in magnitude, not dose-related and/or consistent with normal biological 

variability. 

3.6.2 Clinical chemistry 

(Table 8, Appendix 8 and Annex 3) 

There were no test substance-related effects. 

Minimal increases in total creatine kinase and CK-MM isoenzyme noted in 3/8 females on Study 

Day 21 at 10,000 ppm HFO-1234yf were not considered to be test substance-related because 

they were transient and values were within normal biological variability. 

3.7 Organ weights 

(Table 9, Table 10, Table 11, Appendix 9 and Annex 4) 

HFO-1234yf-related, marginal to slightly higher mean absolute and relative liver weights 

(compared to body and brain weights) were noted at exposure levels ≥ 5,000 ppm in females: 

Text Table 3.7-1: Test substance-related organ weight differences (approximate % difference in 

mean values relative to controls) at Terminal Sacrifice in Females

Sex Exposure Level Females 

HFO-1234yf ppm 0 5,000 10,000 

 No. examined 8 8 8 

Liver Absolute (g) 224 253 272 a

 Absolute % NA 12.9 21.6 a

vs body weight % NA 17.7 a 22.1 a

vs brain weight % NA 18.3 32.8 a

a

Statistically significant difference between mean values for test substance-treated and control groups. 

NA = not applicable: vs = versus 
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Potential morphologic correlates for differences in liver weight were not identified since 

histopathologic evaluation of the liver was not mandated by the protocol. 

Statistically significant difference in heart weight to body weight ratio in the 10,000 ppm group 

males was considered spurious because of the lack of histopathologic findings and the 

discordance between absolute weights and weights relative to body and brain weights. All other 

differences in organ weights were considered incidental and due to normal biological variability. 

3.8 Pathology 

3.8.1 Mortality and macroscopic findings 

(Table 12, Appendix 10 and Annex 5) 

Group 2 male Animal 2147 was euthanized for welfare reasons on Day 16. At necropsy for this 

animal, the Testing Facility documented a prolapse of the rectum; the presence of clear, 

gelatinous material in the colon; and a single thrombus in the left ventricle as well as a single 

thrombus in the right ventricle. There was no evidence to suggest that these findings were 

associated with exposure to the test substance. 

At Day 29, there were a few sporadic macroscopic findings reported by the Testing Facility but 

there were no incidence patterns/trends for these findings to suggest a relationship to test 

substance exposure. 

Macroscopic findings reported included the following: Group 1 male Animal 1145 with diffuse 

red, dark areas of the right cranial lung lobe; Group 1 female Animal 1652 with a few, clear, 

10-19 mm cysts involving the caudal pole of the right kidney; Group 2 female Animal 2648 with 

a few, 10-19 mm, red, dark areas involving the cardiac region of the gastric mucosa; Group 2 

female Animal 2649 with many 2-9 mm dark areas involving all lung lobes; Group 2 female 

Animal 2651 with bilateral dark mandibular lymph nodes; Group 3 male Animal 3148 with 

increased pericardial fluid; Group 3 male Animal 3151 with many small (≤ 1 mm), red, dark 

areas involving all lung lobes; Group 3 male Animal 3152 with focal adhesion of the 

pericardium to the left ventricle and many small (≤ 1 mm), red, dark areas involving the mucosa 

of the lower rectum; Group 3 female Animal 3646 with many 2-9 mm dark areas involving all 

lung lobes; Group 3 female Animal 3648 with bilateral dark mandibular lymph nodes; and 

Group 3 female Animal 3651 with many small (≤ 1 mm), red, dark areas involving all lung lobes 

as well as a few, 10-19 mm, red, dark areas involving the cardiac region of the gastric mucosa 

and one, raised, 2-9 mm dark area of the gastric mucosa of the pylorus. 

3.8.2 Microscopic evaluation 

(Annex 5) 

There were no HFO-1234yf-related microscopic findings involving the heart (left ventricle, right 

ventricle, and septum) and skeletal muscle (rectus femoris, psoas, and soleus muscles as well as 

the diaphragm). 
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Microscopic observations involving the heart were of sporadic incidence and primarily limited to 

cellular infiltration (mononuclear or mixed cell) involving the epicardium, myocardium, and/or 

endocardium. These observations were of minimal, insignificant intensity and showed no 

predilection as to anatomic site. An additional irrelevant finding included the single occurrence 

of focal pigmented macrophages within the epicardium in a single cardiac specimen. 

For skeletal muscle, microscopic observations included sporadic mononuclear cell infiltration, 

neutrophil infiltration, histiocytic infiltration, cytoplasmic basophilia of myofibers, and myofiber 

degeneration involving the rectus femoris, soleus, psoas, and/or diaphragmatic muscle. These 

findings were of limited intensity as they predominantly occurred with minimal degree. A single 

animal, Group 2 female 2647, had minimal to moderate myofiber atrophy involving the rectus 

femoris (right and left), soleus (right and left), and psoas (right and left) muscles that was 

accompanied by slight myofiber mineralization of the left psoas muscle. As these atrophic 

changes were only observed in a single female at the low dose exposure level, they were deemed 

to be unrelated to HFO-1234yf. 

The aforementioned cardiac and skeletal muscle changes were incidental and many were those 

that can be observed in the age of Gottingen minipigs used in this study (McInnes, 2012; 

Svendson et. al., 1998). The microscopic observations showed no consistent patterns/trends 

involving incidence or intensity to suggest a relationship to test substance exposure. 
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4. Conclusion 

Daily inhalation administration of HFO-1234yf to minipigs for 28 days and up to a target 

exposure level of 10,000 ppm (actual achieved exposure level of 10,200 ppm) was not associated 

with cardiotoxicity or skeletal muscle toxicity. The results of this study demonstrate that 

minipigs respond very differently to HFO-1234yf inhalation exposure as compared to rabbits. 

Under the condition of this study, the No-Observed-Adverse-Effect-Level (NOAEL) for 

inhalation toxicity was determined to be the actual achieved exposure level of 10,200 ppm. 
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5. References 

Armitage, P. 1955. Tests for linear trends in proportions and frequencies. Biometrics (Sept.): 

375-386. 

Armitage, P., Berry, G. and Matthews, J.N.S. 2002. Statistical Methods in Medical Research. 

4th edition, p.227. UK: Blackwell Science. 

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

Society, Series A, 160: 268-282. 

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

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

Dunnett, C.W. 1964. New tables for multiple comparisons with a control. Biometrics 20-3: 482-

491. 

Fisher, R.A. 1973. Statistical Methods for Research Workers. 14th edition. New York: Hafner 

Publishing Company. 

Jonckheere, A.R. 1954. A distribution-free k-sample test against ordered alternatives. Biometrics

41: 133-145. 

Kruskal, W.H. and Wallis, W.A. 1952. Use of Ranks in One-Criterion Variance Analysis. 

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

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

American Statistical Association 48: 907-911. 

McInnes, E.F. 2012. Background Lesions in Laboratory Animals: A Color Atlas. Edinburgh: 

Saunders Elsevier. 

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

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

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

15: 560-572. 

Svendson O., Skydsgaard M., Aarup V. and Klastrup S. 1998. Spontaneously Occurring 

Microscopic Lesions in Selected Organs of the Gottingen Minipig. Scand. J. Lab. Anim. Sci. 

Suppl. 1. Vol 25. 

Wilcoxon, F. 1945. Individual comparisons by ranking methods. Biometrics Bulletin 1: 80-83. 
Sponsor reference number: 

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

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

Williams, D.A. 1972. The comparison of several dose levels with a zero dose control. Biometrics

28: 519-531. 
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TABLE 1 Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M P1 Normal Within normal limits 0 4 0

Skin abrasion Dry, Ears 0 2 0

Dry, Ventral Surface 0 3 0

P2 Normal Within normal limits 4 4 0

Skin abrasion Dry, Ears 2 2 0

Dry, Ventral Surface 3 3 0

P3 Normal Within normal limits 4 4 0

Skin abrasion Dry, Ears 2 2 0

Dry, Ventral Surface 3 3 0

40
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M P4 Normal Within normal limits 4 4 0

Skin abrasion Dry, Ears 2 2 0

Dry, Ventral Surface 3 3 0

P5 Normal Within normal limits 8 8 0

P6 Normal Within normal limits 8 8 0

P7 Normal Within normal limits 8 8 0

P8 Normal Within normal limits 8 8 0

P9 Normal Within normal limits 8 8 0

P10 Normal Within normal limits 8 8 0

41
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M P11 Normal Within normal limits 8 7 0

Skin Bruise, HindLimb -Right-Slight 0 1 0

P12 Normal Within normal limits 7 7 0

Skin Bruise, Hindlimb - Right 1 1 0

P13 Normal Within normal limits 8 8 0

P14 Normal Within normal limits 8 8 0

P15 Normal Within normal limits 8 8 0

1 Normal Within normal limits 8 8 0

42
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M 2 Normal Within normal limits 8 8 0

3 Normal Within normal limits 8 7 0

Skin Scab, Hindlimb - Right 0 1 0

4 Normal Within normal limits 7 7 0

Skin Scab, Hindlimb - Right 1 1 0

5 Normal Within normal limits 7 7 0

Skin Scab, Hindlimb - Right 1 1 0

6 Normal Within normal limits 7 7 0

43
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M 6 Skin Scab, Hindlimb - Right 1 1 0

Skin abrasion Dry, Head 0 1 0

7 Normal Within normal limits 7 7 0

Skin Scab, Ears 1 1 0

Scab, Hindlimb - Right 1 1 0

8 Normal Within normal limits 7 7 0

Skin Scab, Ears 1 1 0

Scab, Hindlimb - Right 1 0 0

9 Normal Within normal limits 7 7 0

44
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M 9 Skin Scab, Ears 1 1 0

10 Normal Within normal limits 7 7 0

Skin Scab, Ears 1 1 0

11 Normal Within normal limits 7 7 0

Skin Scab, Ears 1 1 0

12 Normal Within normal limits 7 7 0

Skin Scab, Ears 1 1 0

13 Normal Within normal limits 7 6 0

Discharge Viscous, Red, Penis 0 1 1

Skin Scab, Ears 1 1 0

45
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M 13 Skin Scab, Tail 0 1 0

Ulceration, Tail 1 0 0

14 Normal Within normal limits 6 7 0

Discharge Viscous, Red, Penis 1 0 0

Skin Scab, Ears 1 1 0

Scab, Tail 1 1 0

15 Normal Within normal limits 7 7 0

Skin Scab, Ears 1 1 0

Scab, Tail 1 1 0

16 Normal Within normal limits 5 7 0

46
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M 16 Discharge Aqueous, Red, Penis 1 0 0

Exudate Red Exudate In Cage 1 0 0

Skin Scab, Ears 1 0 0

Scab, Tail 1 0 0

Skin abrasion Dry, Tail 1 1 0

17 Normal Within normal limits 6 6 0

Skin Scab, Ears 1 1 0

Scab, Tail 1 1 0

Skin abrasion Dry, Tail 1 1 0

18 Normal Within normal limits 5 5 0

47
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M 18 Skin Scab, Ears 1 1 0

Scab, Head 1 1 0

Skin abrasion Dry, Tail 1 1 0

19 Normal Within normal limits 6 6 0

Skin Scab, Head 1 1 0

Skin abrasion Dry, Tail 1 1 0

20 Normal Within normal limits 6 6 0

Skin Scab, Hindlimb - Right 1 1 0

Skin abrasion Dry, Tail 1 1 0

21 Normal Within normal limits 6 7 0

48
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M 21 Skin Scab, Hindlimb - Right 1 0 0

Skin abrasion Dry, Ear - Left 1 1 0

22 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ear - Left 1 1 0

23 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ear - Left 1 1 0

24 Normal Within normal limits 8 8 0

25 Normal Within normal limits 8 8 0

26 Normal Within normal limits 8 8 0

49
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1M 27 Normal Within normal limits 8 8 0

28 Normal Within normal limits 8 8 0

29 Normal Within normal limits 8 0 0

2M P1 Normal Within normal limits 0 7 0

Skin abrasion Dry, Ears 0 1 0

P2 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ears 1 1 0

P3 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ears 1 1 0

50
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2M P4 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ears 1 1 0

P5 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ears 1 1 0

P6 Normal Within normal limits 7 8 0

Skin Scab, Ear - Left 1 0 0

P7 Normal Within normal limits 8 8 0

P8 Normal Within normal limits 8 8 0

P9 Normal Within normal limits 8 8 0

51
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2M P10 Normal Within normal limits 8 8 0

P11 Normal Within normal limits 8 8 0

P12 Normal Within normal limits 8 8 0

P13 Normal Within normal limits 8 8 0

P14 Normal Within normal limits 8 8 0

P15 Normal Within normal limits 8 8 0

1 Normal Within normal limits 8 8 0

2 Normal Within normal limits 8 8 0

52
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2M 3 Normal Within normal limits 8 8 0

4 Normal Within normal limits 8 8 0

5 Normal Within normal limits 7 7 0

Skin abrasion Dry, Upper Dorsal Surface 1 1 0

6 Normal Within normal limits 8 8 0

7 Normal Within normal limits 7 7 0

Skin abrasion Dry, Head 1 1 0

8 Normal Within normal limits 7 7 0

Skin abrasion Dry, Head 1 1 0

53
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2M 9 Normal Within normal limits 7 7 0

Skin abrasion Dry, Head 1 1 0

10 Normal Within normal limits 7 7 0

Skin abrasion Dry, Head 1 1 0

11 Normal Within normal limits 8 8 0

12 Normal Within normal limits 8 8 0

13 Normal Within normal limits 8 8 0

14 Normal Within normal limits 7 7 0

Skin abrasion Dry, Head 1 1 0

54
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2M 15 Normal Within normal limits 7 7 0

Skin abrasion Dry, Head 1 1 0

16 Normal Within normal limits 7 7 0

Discharge Aqueous, Red, Anus 1 1 0

Reason for dispatch Prolapsed 0 1 0

17 Normal Within normal limits 6 6 0

Skin abrasion Dry, Hindlimb - Right 1 1 0

18 Normal Within normal limits 5 5 0

Skin Scab, Dorsal surface 1 1 0

55
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2M 18 Skin abrasion Dry, Hindlimb - Right 1 1 0

19 Normal Within normal limits 5 5 0

Skin Scab, Dorsal surface 1 1 0

Skin abrasion Dry, Hindlimb - Right 1 1 0

20 Normal Within normal limits 5 5 0

Skin Scab, Dorsal surface 1 1 0

Skin abrasion Dry, Hindlimb - Right 1 1 0

21 Normal Within normal limits 7 7 0

56
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2M 22 Normal Within normal limits 7 7 0

23 Normal Within normal limits 7 7 0

24 Normal Within normal limits 7 7 0

25 Normal Within normal limits 7 7 0

26 Normal Within normal limits 7 7 0

27 Normal Within normal limits 7 7 0

28 Normal Within normal limits 7 6 0

Skin abrasion Dry, Upper Dorsal Surface 0 1 0

57
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2M 29 Normal Within normal limits 7 0 0

3M P1 Normal Within normal limits 0 7 0

Skin abrasion Dry, Ears 0 1 0

P2 Normal Within normal limits 6 6 0

Skin abrasion Dry, Ears 2 2 0

P3 Normal Within normal limits 6 7 0

Skin abrasion Dry, Ears 2 1 0

P4 Normal Within normal limits 6 6 0

Skin abrasion Dry, Ears 2 2 0

P5 Normal Within normal limits 6 6 0

58
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M P5 Skin abrasion Dry, Ears 2 2 0

P6 Normal Within normal limits 6 6 0

Skin Scab, Ear - Left 2 2 0

P7 Normal Within normal limits 6 6 0

Skin Scab, Ear - Left 2 2 0

P8 Normal Within normal limits 6 6 0

Skin Scab, Ear - Left 2 2 0

P9 Normal Within normal limits 7 7 0

Skin Scab, Ear - Left 1 1 0

59
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M P10 Normal Within normal limits 7 8 0

Skin Scab, Ear - Left 1 0 0

P11 Normal Within normal limits 8 8 0

P12 Normal Within normal limits 8 7 0

Skin abrasion Wet, Ear - Right 0 1 0

P13 Normal Within normal limits 7 8 0

Skin Scab, Ear - Right 1 0 0

P14 Normal Within normal limits 8 7 0

60
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M P14 Skin abrasion Dry, Head 0 1 0

P15 Normal Within normal limits 5 5 0

Skin abrasion Dry, Dorsal surface 1 1 0

Dry, Head 2 2 0

1 Normal Within normal limits 5 5 0

Skin abrasion Dry, Dorsal surface 1 1 0

Dry, Ear - Left 0 1 0

Dry, Head 2 1 0

2 Normal Within normal limits 6 6 0

Skin abrasion Dry, Ear - Left 1 1 0

61
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 2 Skin abrasion Dry, Head 1 1 0

3 Normal Within normal limits 6 6 0

Skin abrasion Dry, Ear - Left 1 1 0

Dry, Head 1 1 0

4 Normal Within normal limits 6 6 0

Skin abrasion Dry, Ear - Left 1 1 1

Dry, Head 1 1 1

Dry, Lower Dorsal Surface 0 0 2

5 Normal Within normal limits 4 4 0

Skin abrasion Dry, Ear - Left 1 1 0

62
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 5 Skin abrasion Dry, Head 3 3 0

Dry, Lower Dorsal Surface 2 2 0

Dry, Upper Dorsal Surface 2 2 0

6 Normal Within normal limits 3 3 0

Skin abrasion Dry, Ear - Left 2 2 0

Dry, Head 3 3 0

Dry, Lower Dorsal Surface 2 2 0

Dry, Upper Dorsal Surface 2 2 0

7 Normal Within normal limits 4 4 0

63
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 7 Skin Scab, Dorsal surface 1 1 0

Scab, Ear - Left 1 1 0

Scab, Lower Dorsal Surface 1 1 0

Skin abrasion Dry, Dorsal surface 1 1 0

Dry, Head 1 1 0

Dry, Lower Dorsal Surface 1 1 0

Dry, Upper Dorsal Surface 2 2 0

8 Normal Within normal limits 4 4 0

Skin Scab, Dorsal surface 1 1 0

64
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 8 Skin Scab, Ear - Left 1 1 0

Scab, Lower Dorsal Surface 1 1 0

Skin abrasion Dry, Dorsal surface 1 1 0

Dry, Head 1 1 0

Dry, Lower Dorsal Surface 1 1 0

Dry, Upper Dorsal Surface 2 2 0

9 Normal Within normal limits 4 4 0

Skin Scab, Dorsal surface 1 1 0

Scab, Ear - Left 1 1 0

65
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 9 Skin Scab, Lower Dorsal Surface 1 1 0

Skin abrasion Dry, Dorsal surface 1 1 0

Dry, Head 1 1 0

Dry, Lower Dorsal Surface 1 1 0

Dry, Upper Dorsal Surface 2 2 0

10 Normal Within normal limits 4 5 0

Skin Scab, Dorsal surface 1 0 0

Scab, Ear - Left 1 1 0

Scab, Lower Dorsal Surface 1 1 0

66
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 10 Skin abrasion Dry, Dorsal surface 1 0 0

Dry, Head 1 1 0

Dry, Lower Dorsal Surface 1 1 0

Dry, Upper Dorsal Surface 2 2 0

11 Normal Within normal limits 6 6 0

Skin Scab, Ear - Left 1 1 0

Scab, Lower Dorsal Surface 1 1 0

Skin abrasion Dry, Lower Dorsal Surface 1 1 0

Dry, Upper Dorsal Surface 1 1 0

67
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 12 Normal Within normal limits 6 6 0

Skin Scab, Ear - Left 1 1 0

Scab, Lower Dorsal Surface 1 1 0

Scab, Upper Dorsal Surface 2 2 0

13 Normal Within normal limits 6 6 0

Skin Scab, Ear - Left 1 1 0

Scab, Lower Dorsal Surface 1 1 0

Scab, Upper Dorsal Surface 2 2 0

68
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 14 Normal Within normal limits 4 4 0

Skin Scab, Dorsal surface 1 1 0

Scab, Ear - Left 1 1 0

Scab, Lower Dorsal Surface 1 1 0

Scab, Upper Dorsal Surface 2 2 0

Skin abrasion Dry, Head 1 1 0

Dry, Upper Dorsal Surface 2 1 0

15 Normal Within normal limits 4 4 0

Skin Scab, Dorsal surface 1 1 0

69
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 15 Skin Scab, Ear - Left 1 1 0

Scab, Lower Dorsal Surface 1 1 0

Scab, Upper Dorsal Surface 2 2 0

Skin abrasion Dry, Head 1 1 0

Dry, Upper Dorsal Surface 1 1 0

16 Normal Within normal limits 7 6 0

Skin Scab, Head 1 1 0

Skin abrasion Dry, Upper Dorsal Surface 0 1 0

17 Normal Within normal limits 6 6 0

70
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 17 Skin Scab, Head 1 1 0

Skin abrasion Dry, Upper Dorsal Surface 1 1 0

18 Normal Within normal limits 4 4 0

Skin Scab, Head 2 2 0

Ulceration, Ear - Left 1 1 0

Skin abrasion Dry, Upper Dorsal Surface 1 1 0

19 Normal Within normal limits 3 3 0

Skin Scab, Head 2 2 0

Scab, Hindlimb - Right 1 1 0

71
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 19 Skin Ulceration, Ear - Left 1 1 0

Skin abrasion Dry, Upper Dorsal Surface 1 1 0

20 Normal Within normal limits 2 2 0

Skin Scab, Head 3 3 0

Scab, Hindlimb - Right 1 1 0

Ulceration, Ear - Left 1 1 0

Skin abrasion Dry, Upper Dorsal Surface 1 1 0

21 Normal Within normal limits 3 3 0

Skin Scab, Ear - Left 0 1 0

72
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 21 Skin Scab, Head 2 2 0

Scab, Hindlimb - Right 1 1 0

Ulceration, Ear - Left 1 0 0

Skin abrasion Dry, Upper Dorsal Surface 1 1 0

22 Normal Within normal limits 4 4 0

Skin Scab, Ear - Left 1 1 0

Scab, Head 1 1 0

Scab, Hindlimb - Right 1 1 0

Skin abrasion Dry, Upper Dorsal Surface 1 1 0

73
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 23 Normal Within normal limits 4 4 0

Skin Scab, Ear - Left 1 1 0

Scab, Head 1 1 0

Scab, Hindlimb - Right 1 1 0

Skin abrasion Dry, Upper Dorsal Surface 1 1 0

24 Normal Within normal limits 7 6 0

Skin Scab, Head 1 1 0

Skin abrasion Dry, Dorsal surface 0 1 0

25 Normal Within normal limits 6 6 0

74
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 25 Skin Scab, Head 1 1 0

Scab, Upper Dorsal Surface 0 1 0

Skin abrasion Dry, Dorsal surface 1 1 0

26 Normal Within normal limits 6 6 0

Skin Scab, Head 1 1 0

Scab, Upper Dorsal Surface 1 1 0

Skin abrasion Dry, Dorsal surface 1 1 0

27 Normal Within normal limits 6 6 0

Skin Scab, Head 1 1 0

75
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3M 27 Skin Scab, Upper Dorsal Surface 1 1 0

Skin abrasion Dry, Dorsal surface 1 1 0

28 Normal Within normal limits 8 8 0

29 Normal Within normal limits 8 0 0

1F P1 Normal Within normal limits 0 7 0

Skin abrasion Dry, Ears 0 1 0

P2 Normal Within normal limits 7 6 0

Skin abrasion Dry, Ears 1 1 0

Dry, Head 0 1 0

76
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1F P3 Normal Within normal limits 6 6 0

Skin abrasion Dry, Ears 1 1 0

Dry, Head 1 1 0

P4 Normal Within normal limits 6 6 0

Skin abrasion Dry, Ears 1 1 0

Dry, Head 1 1 0

P5 Normal Within normal limits 7 7 0

Skin Scab, Head 1 1 0

P6 Normal Within normal limits 8 8 0

P7 Normal Within normal limits 8 8 0

77
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1F P8 Normal Within normal limits 8 8 0

P9 Normal Within normal limits 8 8 0

P10 Normal Within normal limits 8 8 0

P11 Normal Within normal limits 8 8 0

P12 Normal Within normal limits 8 8 0

P13 Normal Within normal limits 8 8 0

P14 Normal Within normal limits 8 8 0

P15 Normal Within normal limits 8 8 0

78
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1F Ra1 Normal Within normal limits 8 8 0

1 Normal Within normal limits 8 8 0

2 Normal Within normal limits 8 8 0

3 Normal Within normal limits 8 8 0

4 Normal Within normal limits 5 5 0

Skin abrasion Dry, Hindlimb - Right 1 1 0

Dry, Upper Dorsal Surface 2 2 0

5 Normal Within normal limits 5 5 0

79
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1F 5 Skin abrasion Dry, Hindlimb - Right 1 1 0

Dry, Upper Dorsal Surface 2 2 0

6 Normal Within normal limits 7 7 0

Skin abrasion Dry, Upper Dorsal Surface 1 1 0

7 Normal Within normal limits 8 8 0

8 Normal Within normal limits 8 8 0

9 Normal Within normal limits 8 8 0

10 Normal Within normal limits 8 6 0

80
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1F 10 Skin abrasion Dry, Head 0 1 0

Dry, Nose 0 1 0

11 Normal Within normal limits 7 7 0

Skin abrasion Dry, Head 1 1 0

12 Normal Within normal limits 8 8 0

13 Normal Within normal limits 8 8 0

14 Normal Within normal limits 8 8 0

15 Normal Within normal limits 8 8 0

16 Normal Within normal limits 7 7 0

81
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1F 16 Skin abrasion Dry, Head 1 1 0

17 Normal Within normal limits 6 7 0

Skin abrasion Dry, Head 1 0 0

Dry, Hindlimb - Right 1 1 0

18 Normal Within normal limits 6 6 0

Skin Scab, Ear - Left 1 1 0

Skin abrasion Dry, Hindlimb - Right 1 1 0

19 Normal Within normal limits 6 6 0

Skin Scab, Ear - Left 1 1 0

82
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1F 19 Skin abrasion Dry, Hindlimb - Right 1 1 0

20 Normal Within normal limits 6 6 0

Skin Scab, Ear - Left 1 1 0

Skin abrasion Dry, Hindlimb - Right 1 1 0

21 Normal Within normal limits 7 7 0

Skin Scab, Ear - Left 1 1 0

22 Normal Within normal limits 7 7 0

Skin Scab, Ear - Left 1 1 0

83
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

1F 23 Normal Within normal limits 8 8 0

24 Normal Within normal limits 8 8 0

25 Normal Within normal limits 8 8 0

26 Normal Within normal limits 8 8 0

27 Normal Within normal limits 8 8 0

28 Normal Within normal limits 8 8 0

29 Normal Within normal limits 8 0 0

2F P1 Normal Within normal limits 0 6 0

84
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F P1 Skin abrasion Dry, Ears 0 1 0

Dry, Ventral Surface 0 1 0

P2 Normal Within normal limits 6 5 0

Skin abrasion Dry, Ears 1 1 0

Dry, Upper Dorsal Surface 0 1 0

Dry, Ventral Surface 1 1 0

P3 Normal Within normal limits 5 5 0

Skin abrasion Dry, Ears 1 1 0

Dry, Upper Dorsal Surface 1 1 0

85
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F P3 Skin abrasion Dry, Ventral Surface 1 1 0

P4 Normal Within normal limits 5 5 0

Skin abrasion Dry, Ears 1 1 0

Dry, Upper Dorsal Surface 1 1 0

Dry, Ventral Surface 1 1 0

P5 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ventral Surface 1 1 0

P6 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ventral Surface 1 1 0

86
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F P7 Normal Within normal limits 8 8 0

P8 Normal Within normal limits 8 8 0

P9 Normal Within normal limits 8 8 0

P10 Normal Within normal limits 8 8 0

P11 Normal Within normal limits 8 8 0

P12 Normal Within normal limits 8 8 0

P13 Normal Within normal limits 8 8 0

P14 Normal Within normal limits 8 8 0

87
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F P15 Normal Within normal limits 8 8 0

Ra1 Normal Within normal limits 8 8 0

1 Normal Within normal limits 8 8 0

2 Normal Within normal limits 8 8 0

3 Normal Within normal limits 8 8 0

4 Normal Within normal limits 8 8 0

5 Normal Within normal limits 7 7 0

Skin abrasion Dry, Hindlimb - Right 1 1 0

88
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F 6 Normal Within normal limits 6 6 0

Skin Scab, Hindlimb - Right 1 1 0

Skin abrasion Dry, Lower Dorsal Surface 1 1 0

7 Normal Within normal limits 8 8 0

8 Normal Within normal limits 8 8 0

9 Normal Within normal limits 8 8 0

10 Normal Within normal limits 8 7 0

Skin abrasion Dry, Upper Ventral Surface 0 1 0

89
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F 10 Skin color Abnormal color, Red, Upper Dorsal Surface 0 1 0

11 Normal Within normal limits 7 7 0

Skin color Abnormal color, Red, Upper Dorsal Surface 1 1 0

12 Normal Within normal limits 8 8 0

13 Normal Within normal limits 8 8 0

14 Normal Within normal limits 8 8 0

15 Normal Within normal limits 8 8 0

16 Normal Within normal limits 8 8 0

90
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F 17 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ear - Left 1 1 0

18 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ear - Left 1 1 0

19 Normal Within normal limits 5 5 0

Skin abrasion Dry, Ear - Left 1 1 0

Dry, Head 2 2 0

20 Normal Within normal limits 5 5 0

Skin abrasion Dry, Dorsal surface 0 1 0

Dry, Ear - Left 1 1 0

91
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F 20 Skin abrasion Dry, Head 2 2 0

21 Normal Within normal limits 5 5 0

Skin abrasion Dry, Dorsal surface 1 1 0

Dry, Ear - Left 1 1 0

Dry, Head 2 2 0

22 Normal Within normal limits 5 5 0

Skin abrasion Dry, Dorsal surface 1 1 0

Dry, Ear - Left 1 1 0

Dry, Head 2 2 0

23 Normal Within normal limits 8 8 0

92
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F 24 Normal Within normal limits 8 8 0

25 Normal Within normal limits 7 7 0

Skin abrasion Dry, Dorsal surface 1 1 0

26 Normal Within normal limits 7 7 0

Skin abrasion Dry, Dorsal surface 1 1 0

27 Normal Within normal limits 7 7 0

Skin abrasion Dry, Dorsal surface 1 1 0

28 Normal Within normal limits 8 8 0

93
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

2F 29 Normal Within normal limits 8 0 0

3F P1 Normal Within normal limits 0 8 0

P2 Normal Within normal limits 8 8 0

P3 Normal Within normal limits 8 8 0

P4 Normal Within normal limits 8 8 0

P5 Normal Within normal limits 8 8 0

P6 Normal Within normal limits 8 8 0

P7 Normal Within normal limits 8 8 0

94
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3F P8 Normal Within normal limits 8 8 0

P9 Normal Within normal limits 8 8 0

P10 Normal Within normal limits 8 8 0

P11 Normal Within normal limits 8 8 0

P12 Normal Within normal limits 8 8 0

P13 Normal Within normal limits 8 8 0

P14 Normal Within normal limits 8 8 0

P15 Normal Within normal limits 8 8 0

95
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3F Ra1 Normal Within normal limits 8 8 0

1 Normal Within normal limits 8 8 0

2 Normal Within normal limits 8 8 0

3 Normal Within normal limits 8 8 0

4 Normal Within normal limits 5 5 0

Skin abrasion Dry, Head 3 3 0

5 Normal Within normal limits 5 5 0

Skin abrasion Dry, Head 3 3 0

Dry, Upper Dorsal Surface 1 1 0

96
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3F 6 Normal Within normal limits 7 7 0

Skin abrasion Dry, Ear - Right 1 1 0

7 Normal Within normal limits 8 8 0

8 Normal Within normal limits 8 8 0

9 Normal Within normal limits 8 8 0

10 Normal Within normal limits 7 7 0

Skin Scab, Lower Dorsal Surface 1 1 0

11 Normal Within normal limits 7 7 0

97
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3F 11 Skin Scab, Lower Dorsal Surface 1 1 0

12 Normal Within normal limits 7 7 0

Skin Scab, Lower Dorsal Surface 1 1 0

13 Normal Within normal limits 7 7 0

Skin Scab, Lower Dorsal Surface 1 1 0

14 Normal Within normal limits 7 7 0

Skin Scab, Lower Dorsal Surface 1 1 0

15 Normal Within normal limits 8 8 0

98
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3F 16 Normal Within normal limits 8 8 0

17 Normal Within normal limits 8 8 0

18 Normal Within normal limits 6 6 0

Skin Scab, Head 1 1 0

Scab, Hindlimb - Right 1 1 0

19 Normal Within normal limits 6 6 0

Skin Scab, Head 1 1 0

Scab, Hindlimb - Right 1 1 0

20 Normal Within normal limits 6 6 0

99
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3F 20 Skin Scab, Head 1 1 0

Scab, Hindlimb - Right 1 1 0

21 Normal Within normal limits 7 7 0

Skin Scab, Head 1 1 0

22 Normal Within normal limits 7 7 0

Skin Scab, Head 1 1 0

23 Normal Within normal limits 8 8 0

24 Normal Within normal limits 8 8 0

25 Normal Within normal limits 8 8 0

100
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 1 (cont) Daily observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation AM PM US PM

3F 26 Normal Within normal limits 8 8 0

27 Normal Within normal limits 8 8 0

28 Normal Within normal limits 8 8 0

29 Normal Within normal limits 8 0 0

101
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

1M 1 Normal Within normal limits 8 8 0 0

2 Normal Within normal limits 8 8 0 0

3 Normal Within normal limits 8 8 0 0

4 Normal Within normal limits 8 8 0 0

5 Normal Within normal limits 8 8 0 0

6 Normal Within normal limits 8 8 0 0

7 Normal Within normal limits 8 8 0 0

8 Normal Within normal limits 8 8 0 0

102
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

1M 9 Normal Within normal limits 8 8 0 0

10 Normal Within normal limits 8 8 0 0

11 Normal Within normal limits 8 8 0 0

12 Normal Within normal limits 8 8 0 0

13 Normal Within normal limits 8 8 0 0

14 Normal Within normal limits 7 7 0 0

15 Normal Within normal limits 8 8 0 0

16 Normal Within normal limits 8 8 0 0

103
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

1M 17 Normal Within normal limits 8 8 0 0

18 Normal Within normal limits 8 8 0 0

19 Normal Within normal limits 8 8 0 0

20 Normal Within normal limits 8 8 0 0

21 Normal Within normal limits 8 8 0 0

22 Normal Within normal limits 8 8 0 0

23 Normal Within normal limits 8 8 0 0

24 Normal Within normal limits 8 8 0 0

104
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

1M 25 Normal Within normal limits 8 8 0 0

26 Normal Within normal limits 8 8 0 0

27 Normal Within normal limits 8 8 0 0

28 Normal Within normal limits 8 8 0 0

2M 1 Normal Within normal limits 7 7 0 0

2 Normal Within normal limits 8 8 0 0

3 Normal Within normal limits 8 8 0 0

4 Normal Within normal limits 8 8 0 0

105
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

2M 5 Normal Within normal limits 8 8 0 0

6 Normal Within normal limits 8 8 0 0

7 Normal Within normal limits 8 8 0 0

8 Normal Within normal limits 8 8 0 0

9 Normal Within normal limits 8 8 0 0

10 Normal Within normal limits 8 8 0 0

11 Normal Within normal limits 8 8 0 0

12 Normal Within normal limits 8 8 0 0

106
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

2M 13 Normal Within normal limits 8 8 0 0

14 Normal Within normal limits 8 8 0 0

15 Normal Within normal limits 8 8 0 0

16 Normal Within normal limits 7 7 0 0

17 Normal Within normal limits 7 7 0 0

18 Normal Within normal limits 7 7 0 0

19 Normal Within normal limits 7 7 0 0

20 Normal Within normal limits 7 7 0 0

107
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

2M 21 Normal Within normal limits 7 7 0 0

22 Normal Within normal limits 7 7 0 0

23 Normal Within normal limits 7 7 0 0

24 Normal Within normal limits 7 7 0 0

25 Normal Within normal limits 7 7 0 0

26 Normal Within normal limits 7 7 0 0

27 Normal Within normal limits 7 7 0 0

28 Normal Within normal limits 7 7 0 0

108
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

3M 1 Normal Within normal limits 8 8 0 0

2 Normal Within normal limits 8 8 0 0

3 Normal Within normal limits 8 8 0 0

4 Normal Within normal limits 8 8 0 0

5 Normal Within normal limits 8 8 0 0

6 Normal Within normal limits 8 8 0 0

7 Normal Within normal limits 8 8 0 0

8 Normal Within normal limits 8 8 0 0

109
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

3M 9 Normal Within normal limits 8 8 0 0

10 Normal Within normal limits 8 8 0 0

11 Normal Within normal limits 8 8 0 0

12 Normal Within normal limits 8 8 0 0

13 Normal Within normal limits 8 8 0 0

14 Normal Within normal limits 8 8 0 0

15 Normal Within normal limits 8 8 0 0

16 Normal Within normal limits 8 8 0 0

110
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

3M 17 Normal Within normal limits 8 8 0 0

18 Normal Within normal limits 8 8 0 0

19 Normal Within normal limits 8 8 0 0

20 Normal Within normal limits 8 8 0 0

21 Normal Within normal limits 8 8 0 0

22 Normal Within normal limits 8 8 0 0

23 Normal Within normal limits 8 8 0 0

24 Normal Within normal limits 8 7 0 0

111
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

3M 24 Muscle reaction Shivering 0 1 0 0

25 Normal Within normal limits 8 8 0 0

26 Normal Within normal limits 8 8 0 0

27 Normal Within normal limits 8 8 0 0

28 Normal Within normal limits 8 8 0 0

1F 1 Normal Within normal limits 8 8 0 0

2 Normal Within normal limits 8 8 0 0

3 Normal Within normal limits 8 8 0 0

4 Normal Within normal limits 8 8 0 0

112
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

1F 5 Normal Within normal limits 8 8 0 0

6 Normal Within normal limits 8 8 0 0

7 Normal Within normal limits 8 8 0 0

8 Normal Within normal limits 8 8 0 0

9 Normal Within normal limits 8 8 0 0

10 Normal Within normal limits 8 8 0 0

11 Normal Within normal limits 8 8 0 0

12 Normal Within normal limits 8 8 0 0

113
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

1F 13 Normal Within normal limits 8 8 0 0

14 Normal Within normal limits 8 8 0 0

15 Normal Within normal limits 8 8 0 0

16 Normal Within normal limits 8 8 0 0

17 Normal Within normal limits 8 8 0 0

18 Normal Within normal limits 8 8 0 0

19 Normal Within normal limits 8 8 0 0

20 Normal Within normal limits 8 8 0 0

114
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

1F 21 Normal Within normal limits 8 8 0 0

22 Normal Within normal limits 8 8 0 0

23 Normal Within normal limits 8 8 0 0

24 Normal Within normal limits 8 8 0 0

25 Normal Within normal limits 8 8 0 0

26 Normal Within normal limits 8 8 0 0

27 Normal Within normal limits 8 8 0 0

28 Normal Within normal limits 8 8 0 0

115
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

2F 1 Normal Within normal limits 8 8 0 0

2 Normal Within normal limits 8 8 0 0

3 Normal Within normal limits 8 8 0 0

4 Normal Within normal limits 8 8 0 0

General condition Laceration, Tail 0 0 0 1

5 Normal Within normal limits 8 8 0 0

6 Normal Within normal limits 8 8 0 0

7 Normal Within normal limits 8 8 0 0

116
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

2F 8 Normal Within normal limits 8 8 0 0

9 Normal Within normal limits 8 8 0 0

10 Normal Within normal limits 8 8 0 0

11 Normal Within normal limits 8 8 0 0

12 Normal Within normal limits 8 8 0 0

13 Normal Within normal limits 8 8 0 0

14 Normal Within normal limits 8 8 0 0

15 Normal Within normal limits 8 8 0 0

117
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

2F 16 Normal Within normal limits 8 8 0 0

17 Normal Within normal limits 8 8 0 0

18 Normal Within normal limits 8 8 0 0

19 Normal Within normal limits 8 8 0 0

20 Normal Within normal limits 8 8 0 0

21 Normal Within normal limits 8 8 0 0

22 Normal Within normal limits 8 8 0 0

23 Normal Within normal limits 8 8 0 0

118
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

2F 24 Normal Within normal limits 8 8 0 0

25 Normal Within normal limits 8 8 0 0

26 Normal Within normal limits 8 8 0 0

27 Normal Within normal limits 8 8 0 0

28 Normal Within normal limits 8 8 0 0

3F 1 Normal Within normal limits 8 8 0 0

General condition Laceration, Tail 0 0 1 0

2 Normal Within normal limits 7 7 0 0

119
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

3F 2 General condition Laceration, Tail 1 1 0 0

3 Normal Within normal limits 8 8 0 0

4 Normal Within normal limits 8 8 0 0

5 Normal Within normal limits 8 8 0 0

6 Normal Within normal limits 8 8 0 0

7 Normal Within normal limits 8 8 0 0

8 Normal Within normal limits 8 8 0 0

9 Normal Within normal limits 8 8 0 0

120
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

3F 10 Normal Within normal limits 8 8 0 0

11 Normal Within normal limits 8 8 0 0

Breathing Coughing 0 0 1 0

12 Normal Within normal limits 8 8 0 0

13 Normal Within normal limits 8 8 0 0

14 Normal Within normal limits 8 8 0 0

15 Normal Within normal limits 8 8 0 0

16 Normal Within normal limits 8 8 0 0

17 Normal Within normal limits 8 8 0 0

121
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

3F 18 Normal Within normal limits 8 8 0 0

19 Normal Within normal limits 8 8 0 0

20 Normal Within normal limits 8 8 0 0

21 Normal Within normal limits 8 8 0 0

22 Normal Within normal limits 8 8 0 0

23 Normal Within normal limits 8 8 0 0

24 Normal Within normal limits 8 8 0 0

25 Normal Within normal limits 8 8 0 0

122
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 2 (cont) Dose observations - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Number of animals affected

/Sex Day Category Observation 2 Hours 4 Hours US1 US2

3F 26 Normal Within normal limits 8 8 0 0

27 Normal Within normal limits 8 8 0 0

28 Normal Within normal limits 8 8 0 0

123
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 3 Clinical signs - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Number of animals affected

Category Observation Week Group/Sex: 1M 2M 3M 1F 2F 3F

Initial no: 8 8 8 8 8 8

Normal Within normal limits P2 8 8 8 8 8 8

P3 8 8 5 8 8 8

1 7 7 4 8 8 8

2 6 7 4 8 8 7

3 7 6 3 8 6 7

4 8 7 8 8 8 8

Discharge Viscous, Red, Penis 2 1 0 0 0 0 0

Skin Scab, Dorsal surface 1 0 0 1 0 0 0

2 0 0 1 0 0 0

3 0 0 1 0 0 0

Scab, Ear - Left 1 0 0 1 0 0 0

2 0 0 1 0 0 0

3 0 0 1 0 0 0

Scab, Ears 1 1 0 0 0 0 0

2 1 0 0 0 0 0

3 1 0 0 0 0 0

124
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 3 (cont) Clinical signs - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Number of animals affected

Category Observation Week Group/Sex: 1M 2M 3M 1F 2F 3F

Initial no: 8 8 8 8 8 8

Skin Scab, Hindlimb - Right 1 1 0 0 0 0 0

2 1 0 0 0 0 0

3 1 0 0 0 0 0

Scab, Lower Dorsal Surface 1 0 0 1 0 0 0

2 0 0 1 0 0 1

3 0 0 1 0 0 0

Scab, Upper Dorsal Surface 1 0 0 1 0 0 0

2 0 0 1 0 0 0

3 0 0 1 0 0 0

Ulceration, Ear - Left 3 0 0 1 0 0 0

Skin abrasion Dry, Dorsal surface P3 0 0 1 0 0 0

1 0 0 1 0 0 0

2 0 0 1 0 0 0

3 0 0 1 0 0 0

Dry, Ear - Left 3 0 1 1 0 2 0

125
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 3 (cont) Clinical signs - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Number of animals affected

Category Observation Week Group/Sex: 1M 2M 3M 1F 2F 3F

Initial no: 8 8 8 8 8 8

Skin abrasion Dry, Head P3 0 0 2 0 0 0

1 0 1 1 0 0 0

2 0 1 1 0 0 0

3 0 1 1 0 0 1

Dry, Lower Dorsal Surface 1 0 0 1 0 0 0

2 0 0 1 0 0 0

3 0 0 1 0 0 0

Dry, Upper Dorsal Surface 1 0 0 1 0 0 0

2 0 0 1 0 0 0

3 0 0 1 0 0 0

126
Sponsor refe

Huntingdo

TABLE 4 Body weights - group mean values (kg)  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Week Week Change Change 

/Sex P1 P2 P3 1 2 3 4 P3-1 P3-2 

Statistics test Av Av Av Wi Wi Wi Wi Wi Wi 

1M Mean 6.8 7.2 7.7 8.6 9.3 10.3 10.9 0.9 1.7 

 SD 0.85 0.84 0.90 0.99 1.11 1.19 1.31 0.15 0.36 

N 8 8 8 8 8 8 8 8 8 

2M Mean 6.3 6.7 7.3 8.1 8.8 9.6 10.2 0.8 1.5 

 SD 0.70 0.77 0.94 1.06 1.16 1.11 1.09 0.27 0.34 

N 8 8 8 8 8 7 7 8 8 

3M Mean 5.9 6.5 7.1 7.8 8.6 9.6 9.9 0.7 1.5 

 SD 0.83 0.89 1.01 1.20 1.37 1.62 1.70 0.25 0.49 

N 8 8 8 8 8 8 8 8 8 

127
Sponsor refe

Huntingdo

TABLE 4 (cont) Body weights - group mean values (kg)  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Week Week Change Change 

/Sex P1 P2 P3 1 2 3 4 P3-1 P3-2 

Statistics test Av Av Av Wi Wi Wi Wi Wi Wi 

1F Mean 6.8 7.5 8.1 9.2 10.0 11.0 11.7 1.1 1.9 

 SD 0.40 0.52 0.68 0.89 0.97 1.07 1.09 0.24 0.35 

N 8 8 8 8 8 8 8 8 8 

2F Mean 6.4 7.0 7.6 8.7 9.4 10.7 11.5 1.1 1.8 

 SD 1.05 1.16 1.25 1.34 1.48 1.65 1.65 0.22 0.41 

N 8 8 8 8 8 8 8 8 8 

3F Mean 6.7 7.2 7.8 8.9 9.8 11.0 11.7 1.1 1.9 

 SD 0.54 0.74 0.85 0.97 1.12 1.36 1.51 0.20 0.33 

N 8 8 8 8 8 8 8 8 8 

128
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 5 Body weight change - group mean values (kg)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Week Week

/Sex P1-P2 P2-P3 P3-1 1-2 2-3 3-4

Statisticstest Av Av Wi Wi Wi Wi

1M Mean 0.5 0.5 0.9 0.8 1.0 0.5

SD 0.34 0.19 0.15 0.25 0.30 0.23

N 8 8 8 8 8 8

2M Mean 0.4 0.6 0.8 0.8 1.0 0.6

SD 0.25 0.25 0.27 0.17 0.17 0.08

N 8 8 8 8 7 7

3M Mean 0.5 0.6 0.7 0.8 0.9 0.3**

SD 0.20 0.24 0.25 0.28 0.47 0.12

N 8 8 8 8 8 8

129
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 5 (cont) Body weight change - group mean values (kg)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Week Week

/Sex P1-P2 P2-P3 P3-1 1-2 2-3 3-4

Statisticstest Av Av Wi Wi Wi Wi

1F Mean 0.6 0.7 1.1 0.7 1.0 0.7

SD 0.26 0.24 0.24 0.16 0.18 0.14

N 8 8 8 8 8 8

2F Mean 0.5 0.6 1.1 0.7 1.3 0.7

SD 0.27 0.14 0.22 0.28 0.22 0.27

N 8 8 8 8 8 8

3F Mean 0.5 0.6 1.1 0.9 1.2 0.7

SD 0.31 0.20 0.20 0.18 0.33 0.26

N 8 8 8 8 8 8

130
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 6 Visual food consumption - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Number of animals affected

Category Observation Day Group/Sex: 1M 2M 3M 1F 2F 3F

Initial no: 8 8 8 8 8 8

Visual Food 

Consumption 4: All food consumed P8 8 8 8 8 8 8

P9 8 8 8 8 8 8

P10 8 8 8 8 8 8

P11 8 8 8 8 8 8

P12 8 8 8 8 8 8

P13 8 8 8 8 8 8

P14 8 8 8 8 8 8

P15 8 8 8 8 8 8

Ra1 0 0 0 8 8 8

1 8 8 8 8 8 8

2 8 8 8 8 8 8

3 8 8 8 8 8 8

4 8 8 8 8 8 8

5 8 8 8 8 8 8

6 8 8 8 8 8 8

7 8 8 8 8 8 8

8 8 8 8 8 8 8

9 8 8 8 8 8 8

10 8 8 8 8 8 8

11 8 8 8 8 8 8

12 8 8 8 8 8 8

13 8 8 8 8 8 8

14 8 8 8 8 8 8

15 8 8 8 8 8 8

131
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 6 (cont) Visual food consumption - group distribution of observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Number of animals affected

Category Observation Day Group/Sex: 1M 2M 3M 1F 2F 3F

Initial no: 8 8 8 8 8 8

Visual Food 

Consumption 4: All food consumed 16 8 7 8 8 8 8

17 8 7 8 8 8 8

18 8 7 8 8 8 8

19 8 7 8 8 8 8

20 8 7 8 8 8 8

21 8 7 8 8 8 8

22 8 7 8 8 8 8

23 8 7 8 8 8 8

24 8 7 8 8 8 8

25 8 7 8 8 8 8

26 8 7 8 8 8 8

27 8 7 8 8 8 8

28 8 7 8 8 8 8

132
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 Hematology - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Pretest 1 g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

Mean 12.4 38.5 7.69 269.9 627 50.5 16.2 32.2 17.4 13.57

SD 0.88 3.39 1.001 38.01 197.2 5.76 1.91 0.65 0.92 2.797

N 8 8 8 8 8 8 8 8 8 8

Group 2

Mean 11.6 36.2 7.40 249.5 636 49.1 15.7 31.9 18.5 11.97

SD 1.13 3.00 0.766 51.02 118.4 2.51 1.11 0.75 0.79 2.307

N 8 8 8 8 8 8 8 8 8 8

Group 3

Mean 12.0 37.3 7.62 261.6 666 49.3 15.8 32.0 18.8* 12.71

SD 0.89 2.80 0.662 70.46 180.5 5.93 1.94 0.60 1.52 1.924

N 8 8 8 8 8 8 8 8 8 8

133
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion ANEU ALYM AMONO AEOS ABASO ALUC

Pretest 1 x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

Mean 5.46 7.48 0.29 0.12 0.02 0.19

SD 2.568 2.044 0.112 0.043 0.006 0.102

N 8 8 8 8 8 8

Group 2

Mean 4.66 6.63 0.26 0.17 0.02 0.24

SD 0.919 1.933 0.090 0.065 0.009 0.105

N 8 8 8 8 8 8

Group 3

Mean 4.65 7.44 0.24 0.14 0.02 0.22

SD 1.144 1.062 0.063 0.041 0.005 0.083

N 8 8 8 8 8 8

134
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Pretest 2 g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

Mean 12.8 38.7 7.85 202.3 579 49.7 16.4 33.0 17.1 11.51

SD 0.81 2.33 0.659 44.62 171.2 5.72 2.04 0.71 0.92 1.976

N 8 8 8 8 8 8 8 8 8 8

Group 2

Mean 11.7 36.0 7.48 203.6 589 48.3 15.6 32.4 18.1 9.62

SD 1.17 3.21 0.738 78.34 165.5 3.15 1.17 0.71 0.90 1.683

N 8 8 8 8 8 8 8 8 8 8

Group 3

Mean 12.1 37.9 7.62 301.2 618 50.2 16.1 32.0* 19.0* 10.73

SD 1.27 3.53 0.787 173.25 126.3 6.15 1.99 0.68 2.57 2.135

N 8 8 8 8 8 8 8 8 8 8

135
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion ANEU ALYM AMONO AEOS ABASO ALUC

Pretest 2 x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

Mean 5.02 5.96 0.23 0.11 0.01 0.19

SD 1.473 1.393 0.091 0.042 0.007 0.042

N 8 8 8 8 8 8

Group 2

Mean 3.65 5.45 0.19 0.13 0.01 0.19

SD 0.462 1.540 0.064 0.088 0.006 0.041

N 8 8 8 8 8 8

Group 3

Mean 4.47 5.80 0.19 0.09 0.01 0.17

SD 2.005 0.729 0.028 0.019 0.007 0.043

N 8 8 8 8 8 8

136
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Term Day 21 g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

Mean 13.0 39.1 7.92 203.5 506 49.7 16.5 33.2 17.2 10.87

SD 0.87 2.79 0.643 84.76 152.3 5.27 1.76 0.82 1.94 4.092

N 8 8 8 8 8 8 8 8 8 8

Group 2

Mean 12.2 36.9 7.99 144.3 515 46.2 15.2 32.8 17.3 8.52

SD 1.15 2.61 0.608 70.22 87.5 2.56 1.19 1.03 1.03 1.927

N 7 7 7 7 7 7 7 7 7 7

Group 3

Mean 12.8 38.2 8.02 129.6 499 48.1 16.0 33.3 17.1 10.70

SD 1.26 3.54 0.900 52.60 147.9 5.50 1.90 0.39 1.34 2.027

N 8 8 8 8 8 8 8 8 8 8

137
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion ANEU ALYM AMONO AEOS ABASO ALUC

Term Day 21 x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

Mean 4.40 5.90 0.24 0.14 0.03 0.15

SD 2.867 1.745 0.075 0.077 0.056 0.069

N 8 8 8 8 8 8

Group 2

Mean 2.98 4.91 0.26 0.18 0.01 0.19

SD 0.932 1.290 0.078 0.055 0.005 0.066

N 7 7 7 7 7 7

Group 3

Mean 4.44 5.63 0.26 0.17 0.02 0.19

SD 1.382 0.974 0.053 0.043 0.025 0.074

N 8 8 8 8 8 8

138
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Term Day 28 g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

Mean 13.8 41.7 8.37 164.5 496 50.1 16.6 33.1 17.4 9.76

SD 1.09 3.67 0.866 55.13 161.7 4.64 1.73 0.73 1.35 1.829

N 8 8 8 8 8 8 8 8 8 8

Group 2

Mean 12.9 39.0 8.35 162.8 519 46.8 15.5 33.1 17.9 8.41

SD 0.82 2.32 0.669 50.60 115.8 2.48 1.17 1.01 1.30 2.143

N 7 7 7 7 7 7 7 7 7 7

Group 3

Mean 13.4 40.7 8.37 138.4 520 49.1 16.1 32.7 17.4 10.90

SD 1.08 3.08 0.849 52.01 113.4 5.26 1.94 0.48 1.32 2.197

N 8 8 8 8 8 8 8 8 8 8

139
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion ANEU ALYM AMONO AEOS ABASO ALUC

Term Day 28 x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

Mean 3.54 5.65 0.29 0.22 0.02 0.05

SD 1.038 1.277 0.057 0.086 0.005 0.020

N 8 8 8 8 8 8

Group 2

Mean 2.83 5.06 0.29 0.17 0.01** 0.04

SD 0.778 1.743 0.069 0.037 0.005 0.014

N 7 7 7 7 7 7

Group 3

Mean 4.33 5.93 0.34 0.23 0.02 0.06

SD 1.711 1.186 0.063 0.054 0.005 0.026

N 8 8 8 8 8 8

140
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Pretest 1 g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

Mean 13.1 40.5 7.98 241.4 643 51.2 16.5 32.3 17.1 13.69

SD 0.75 2.23 0.688 75.55 100.8 5.82 2.05 0.55 1.24 1.840

N 8 8 8 8 8 8 8 8 8 8

Group 2

Mean 12.6 38.8 7.84 304.2 625 49.8 16.1 32.5 17.9 11.15

SD 0.68 2.40 0.624 60.02 137.4 5.71 1.82 0.62 1.22 2.223

N 8 8 8 8 8 8 8 8 8 8

Group 3

Mean 12.6 38.6 7.96 250.2 585 48.5 15.9 32.8 18.1 14.28

SD 1.01 2.77 0.581 53.68 124.0 3.41 1.44 0.86 1.00 3.102

N 8 8 8 8 8 8 8 8 8 8

141
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion ANEU ALYM AMONO AEOS ABASO ALUC

Pretest 1 x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

Mean 3.12 9.79 0.23 0.20 0.04 0.31

SD 1.039 1.562 0.101 0.095 0.050 0.056

N 8 8 8 8 8 8

Group 2

Mean 2.68 7.72 0.28 0.19 0.02 0.27

SD 0.870 1.772 0.092 0.061 0.005 0.106

N 8 8 8 8 8 8

Group 3

Mean 4.44 9.03 0.37* 0.21 0.03 0.21*

SD 2.922 2.304 0.138 0.093 0.027 0.100

N 8 8 8 8 8 8

142
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Pretest 2 g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

Mean 12.7 39.4 7.76 204.6 660 51.1 16.5 32.2 17.0 11.23

SD 0.65 2.37 0.674 82.18 80.7 5.52 2.00 0.88 1.25 1.320

N 8 8 8 8 8 8 8 8 8 8

Group 2

Mean 12.3 37.6 7.62 256.1 641 49.4 16.1 32.6 18.0 8.99

SD 1.08 3.22 0.254 59.24 110.5 5.14 1.66 0.34 1.69 1.940

N 8 8 8 8 8 8 8 8 8 8

Group 3

Mean 12.7 39.4 8.01 286.3 575 49.4 15.9 32.1 18.5 10.51

SD 1.10 3.15 0.669 84.41 119.3 3.84 1.45 0.69 1.10 1.776

N 8 8 8 8 8 8 8 8 8 8

143
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion ANEU ALYM AMONO AEOS ABASO ALUC

Pretest 2 x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

Mean 2.55 8.07 0.22 0.12 0.02 0.25

SD 0.852 1.278 0.063 0.056 0.019 0.065

N 8 8 8 8 8 8

Group 2

Mean 1.90 6.58 0.23 0.10 0.02 0.17

SD 0.885 1.558 0.097 0.050 0.018 0.077

N 8 8 8 8 8 8

Group 3

Mean 2.18 7.79 0.22 0.10 0.04 0.18

SD 0.531 1.675 0.110 0.041 0.043 0.082

N 8 8 8 8 8 8

144
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Term Day 21 g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

Mean 13.5 40.3 8.05 167.4 561 50.3 16.8 33.4 16.6 10.33

SD 0.73 2.17 0.679 82.96 89.1 5.38 2.00 0.73 1.70 1.146

N 8 8 8 8 8 8 8 8 8 8

Group 2

Mean 13.1 38.8 7.99 176.8 474 48.8 16.4 33.7 17.2 9.74

SD 1.00 3.23 0.432 69.86 159.4 5.08 1.73 0.59 1.22 1.648

N 8 8 8 8 8 8 8 8 8 8

Group 3

Mean 13.2 40.6 8.12 209.3 515 50.2 16.4 32.5 18.5* 11.61

SD 1.30 3.47 0.559 39.06 87.4 4.96 1.89 1.24 1.03 1.601

N 8 8 8 8 8 8 8 8 8 8

145
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion ANEU ALYM AMONO AEOS ABASO ALUC

Term Day 21 x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

Mean 2.01 7.61 0.23 0.23 0.03 0.23

SD 0.445 0.749 0.080 0.113 0.034 0.105

N 8 8 8 8 8 8

Group 2

Mean 2.46 6.69 0.23 0.20 0.01 0.15

SD 0.448 1.710 0.098 0.067 0.004 0.082

N 8 8 8 8 8 8

Group 3

Mean 3.14** 7.79 0.23 0.25 0.03 0.16

SD 1.140 1.412 0.094 0.138 0.015 0.079

N 8 8 8 8 8 8

146
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Term Day 28 g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

Mean 13.4 40.4 7.93 178.4 561 51.3 17.0 33.2 17.6 10.76

SD 1.05 3.39 0.907 77.35 88.0 5.42 2.14 1.21 2.32 1.470

N 8 8 8 8 8 8 8 8 8 8

Group 2

Mean 12.9 38.8 7.92 136.2 576 49.1 16.3 33.2 17.2 9.64

SD 0.83 2.20 0.379 42.66 79.5 4.78 1.70 0.55 0.79 1.579

N 8 8 8 8 8 8 8 8 8 8

Group 3

Mean 13.3 40.5 8.05 182.1 503 50.4 16.6 32.8 18.7 11.04

SD 1.33 3.41 0.617 41.67 117.0 4.92 1.98 1.21 1.28 2.308

N 8 8 8 8 8 8 8 8 8 8

147
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 7 (cont) Hematology - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion ANEU ALYM AMONO AEOS ABASO ALUC

Term Day 28 x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

Mean 2.30 7.73 0.34 0.29 0.03 0.07

SD 0.426 1.199 0.138 0.122 0.019 0.031

N 8 8 8 8 8 8

Group 2

Mean 2.36 6.71 0.26 0.25 0.02 0.05

SD 0.592 1.531 0.053 0.065 0.008 0.025

N 8 8 8 8 8 8

Group 3

Mean 3.08 7.30 0.25 0.32 0.04 0.05

SD 1.154 1.975 0.043 0.105 0.026 0.022

N 8 8 8 8 8 8

148
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 Clinical chemistry - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion AST ALT CK

Pretest 1 U/L U/L U/L

Group 1

Mean 26 47 378

SD 4.6 5.8 203.4

N 8 8 8

Group 2

Mean 32 40 681

SD 17.7 4.0 769.6

N 8 8 8

Group 3

Mean 33 45 741

SD 9.2 6.7 698.9

N 8 8 8

149
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Pretest 1 U/L % U/L % U/L NG/mL NG/mL

Group 1

Mean 387 63.8 248 11.9 40.5 26.2 0.0

SD 215.1 13.62 152.8 4.86 21.92 13.89 0.00

N 8 8 8 8 8 8 8

Group 2

Mean 665 50.5 415 13.7 76.4 79.0 0.0

SD 740.1 17.54 569.0 4.52 65.73 140.13 0.00

N 8 8 8 8 8 8 8

Group 3

Mean 716 42.6* 389 13.0 75.2 64.3 0.0

SD 651.7 16.72 484.1 6.36 49.21 77.79 0.00

N 8 8 8 8 8 8 8

150
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion AST ALT CK

Pretest 2 U/L U/L U/L

Group 1

Mean 38 45 630

SD 6.6 4.1 382.0

N 8 8 8

Group 2

Mean 30 38* 589

SD 12.4 4.8 524.4

N 8 8 8

Group 3

Mean 36 46 578

SD 7.9 5.7 528.4

N 8 8 8

151
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Pretest 2 U/L % U/L % U/L NG/mL NG/mL

Group 1

Mean 617 53.7 375 16.0 96.6 34.0 0.0

SD 373.3 21.92 320.2 5.88 73.10 31.19 0.00

N 8 8 8 8 8 8 8

Group 2

Mean 549 55.7 338 11.4 57.3 47.0 0.0

SD 476.1 9.64 352.7 2.46 42.28 91.23 0.00

N 8 8 8 8 8 8 8

Group 3

Mean 539 51.0 283 12.4 63.2 29.1 0.0

SD 483.8 6.79 272.6 3.24 48.65 24.26 0.00

N 8 8 8 8 8 8 8

152
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion AST ALT CK

Term Day 21 U/L U/L U/L

Group 1

Mean 29 43 424

SD 4.8 7.0 221.8

N 8 8 8

Group 2

Mean 26 36 302

SD 4.5 3.6 57.3

N 7 7 7

Group 3

Mean 32 44 684

SD 12.9 5.4 1026.7

N 8 8 8

153
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Term Day 21 U/L % U/L % U/L NG/mL NG/mL

Group 1

Mean 402 53.5 244 11.8 38.9 36.0 0.0

SD 212.6 20.51 214.1 5.49 12.88 29.20 0.00

N 8 8 8 8 8 8 8

Group 2

Mean 283 62.2 185 11.8 32.6 25.3 0.0

SD 51.8 27.45 95.8 3.49 8.86 13.72 0.00

N 7 7 7 7 7 7 7

Group 3

Mean 612 73.8 524 10.0 32.7 57.6 0.0

SD 875.9 17.24 846.3 4.50 9.94 88.76 0.01

N 8 8 8 8 8 8 8

154
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion AST ALT CK

Term Day 28 U/L U/L U/L

Group 1

Mean 28 43 301

SD 8.4 5.8 83.0

N 8 8 8

Group 2

Mean 26 38 532

SD 4.7 4.2 569.2

N 7 7 7

Group 3

Mean 29 44 660

SD 8.8 7.6 768.3

N 8 8 8

155
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Term Day 28 U/L % U/L % U/L NG/mL NG/mL

Group 1

Mean 286 27.7 80 16.7 48.1 25.0 0.0

SD 77.7 12.52 40.3 4.45 19.15 27.06 0.00

N 8 8 8 8 8 8 8

Group 2

Mean 490 36.8 231 13.0 57.1 43.4 0.0

SD 502.5 14.46 348.8 3.26 44.36 42.21 0.00

N 7 7 7 7 7 7 7

Group 3

Mean 598 38.8 322 11.8* 58.5 50.2 0.0

SD 671.5 16.41 491.3 4.64 59.44 52.06 0.01

N 8 8 8 8 8 8 8

156
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion AST ALT CK

Pretest 1 U/L U/L U/L

Group 1

Mean 27 42 544

SD 4.8 5.6 201.8

N 8 8 8

Group 2

Mean 76 58 692

SD 127.6 52.2 447.4

N 8 8 7

Group 3

Mean 33 44 854

SD 14.1 11.4 432.6

N 8 8 8

157
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Pretest 1 U/L % U/L % U/L NG/mL NG/mL

Group 1

Mean 573 63.6 383 12.5 66.3 21.4 0.2

SD 187.1 18.82 231.2 4.70 23.86 12.52 0.59

N 8 8 8 8 8 8 8

Group 2

Mean 6559 78.5 6224 11.0 168.9 77.4 0.0

SD 16900.2 11.94 16349.0 6.15 305.58 96.22 0.11

N 8 8 8 8 8 8 8

Group 3

Mean 849 78.5 680 10.6 81.4 28.7 0.0

SD 414.3 10.31 376.7 5.89 37.20 17.92 0.00

N 8 8 8 8 8 8 8

158
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion AST ALT CK

Pretest 2 U/L U/L U/L

Group 1

Mean 26 41 1432

SD 5.1 4.8 2433.5

N 8 8 8

Group 2

Mean 49 54 618

SD 64.7 45.3 591.2

N 8 8 7

Group 3

Mean 27 46 705

SD 5.2 12.6 288.3

N 8 8 8

159
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Pretest 2 U/L % U/L % U/L NG/mL NG/mL

Group 1

Mean 1163 71.9 972 8.9 68.6 44.0 0.3

SD 1751.8 18.97 1722.6 5.67 52.20 45.14 0.89

N 8 8 8 8 8 8 8

Group 2

Mean 2723 66.9 2412 10.2 152.2 146.3 0.0

SD 6091.9 20.29 5803.1 3.63 245.17 346.39 0.06

N 8 8 8 8 8 8 8

Group 3

Mean 652 68.8 455 13.5 78.4 45.7 0.0

SD 270.5 16.12 256.7 5.28 27.16 26.62 0.00

N 8 8 8 8 8 8 8

160
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion AST ALT CK

Term Day 21 U/L U/L U/L

Group 1

Mean 28 40 1055

SD 8.3 3.7 532.1

N 8 8 8

Group 2

Mean 29 60 1683

SD 18.6 63.4 1898.4

N 8 8 7

Group 3

Mean 23 42 1063

SD 3.7 7.3 690.5

N 8 8 5

161
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Term Day 21 U/L % U/L % U/L NG/mL NG/mL

Group 1

Mean 1089 91.0 1002 5.8 56.1 31.1 0.5

SD 446.0 6.82 433.3 3.44 27.31 37.00 1.39

N 8 8 8 8 8 8 8

Group 2

Mean 3774 86.1 3497 8.8 184.8 33.0 0.0

SD 6350.6 10.10 5958.4 5.46 255.78 39.36 0.03

N 8 8 8 8 8 8 8

Group 3

Mean 5854 61.9* 5174 9.5 276.1* 30.7 0.0

SD 7066.2 28.15 6800.8 5.99 266.83 17.40 0.00

N 8 8 8 8 8 8 8

162
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion AST ALT CK

Term Day 28 U/L U/L U/L

Group 1

Mean 25 41 787

SD 4.9 4.0 373.2

N 8 8 8

Group 2

Mean 40 59 473

SD 49.1 64.7 238.8

N 8 8 7

Group 3

Mean 29 41 1214

SD 12.7 9.7 1054.9

N 8 8 8

163
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 8 (cont) Clinical chemistry - group mean values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Term Day 28 U/L % U/L % U/L NG/mL NG/mL

Group 1

Mean 726 47.8 370 15.0 110.6 41.5 0.5

SD 343.3 10.96 228.0 5.46 74.33 38.54 1.46

N 8 8 8 8 8 8 8

Group 2

Mean 5177 37.4 4725 9.7* 154.2 45.8 0.0

SD 13397.5 25.75 12974.7 4.21 309.87 53.54 0.03

N 8 8 8 8 8 8 8

Group 3

Mean 1056 40.5 524 8.6** 87.7 167.3 0.0

SD 857.2 16.56 580.5 2.28 61.39 228.95 0.00

N 8 8 8 8 8 8 8

164
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 9 Organ weights - group mean absolute values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group/ 

Sex

Terminal 

Body weight Brain Heart Kidneys Liver

Lungs and 

Bronchi

Statisticstest Wi Wi Wi Wi Wi Wi

1M Mean 10750.0 57.217 60.056 71.203 234.307 89.734

SD 1331.0 3.040 11.035 11.046 41.527 14.197

N 8 8 8 8 8 8

2M Mean 10214.3 58.078 59.300 63.474 222.538 88.017

SD 1009.0 6.629 6.161 11.811 50.518 7.733

N 7 7 7 7 7 7

3M Mean 9850.0 56.548 61.060 72.587 232.517 83.012

SD 1725.4 3.309 9.540 13.441 57.898 13.276

N 8 8 8 8 8 8

165
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 9 (cont) Organ weights - group mean absolute values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group/ 

Sex

Terminal 

Body weight Brain Heart Kidneys Liver

Lungs and 

Bronchi

Statisticstest Wi Wi Wi Wi Wi Wi

1F Mean 11571.3 60.711 50.808 53.208 223.714 83.071

SD 1212.6 4.339 7.837 7.299 31.323 8.011

N 8 8 8 8 8 8

2F Mean 11083.8 57.416 49.035 52.355 252.527 83.066

SD 1581.9 3.493 4.627 8.533 48.929 10.424

N 8 8 8 8 8 7

3F Mean 11470.0 55.358* 51.797 52.664 272.062* 86.884

SD 1442.7 4.127 6.670 11.254 48.408 12.399

N 8 8 8 8 8 8

166
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 10 Organ weights - group mean values relative (%) to body weight (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group/ 

Sex

Terminal 

Body weight Brain Heart Kidneys Liver

Lungs and 

Bronchi

Statisticstest Wi Wi Wi Wi Wi Wi

1M Mean 10750.0 0.538 0.556 0.669 2.18 0.834

SD 1331.0 0.052 0.043 0.109 0.25 0.064

N 8 8 8 8 8 8

2M Mean 10214.3 0.569 0.583 0.620 2.17 0.866

SD 1009.0 0.037 0.054 0.084 0.37 0.075

N 7 7 7 7 7 7

3M Mean 9850.0 0.586 0.623* 0.739 2.34 0.851

SD 1725.4 0.082 0.047 0.072 0.21 0.108

N 8 8 8 8 8 8

167
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 10 (cont) Organ weights - group mean values relative (%) to body weight (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group/ 

Sex

Terminal 

Body weight Brain Heart Kidneys Liver

Lungs and 

Bronchi

Statisticstest Wi Wi Wi Wi Wi Wi

1F Mean 11571.3 0.529 0.440 0.459 1.93 0.721

SD 1212.6 0.056 0.054 0.037 0.20 0.061

N 8 8 8 8 8 8

2F Mean 11083.8 0.524 0.446 0.474 2.28** 0.742

SD 1581.9 0.053 0.042 0.055 0.29 0.067

N 8 8 8 8 8 7

3F Mean 11470.0 0.488 0.458 0.456 2.36** 0.762

SD 1442.7 0.058 0.084 0.062 0.17 0.093

N 8 8 8 8 8 8

168
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 11 Organ weights - group mean values relative (%) to brain weight (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group/ 

Sex Brain Heart Kidneys Liver

Lungs and 

Bronchi

Statisticstest Wi Wi Wi Wi Wi

1M Mean 57.217 105 125 409 156

SD 3.040 17 20 65 20

N 8 8 8 8 8

2M Mean 58.078 103 109 383 153

SD 6.629 12 16 73 16

N 7 7 7 7 7

3M Mean 56.548 108 128 410 147

SD 3.309 14 23 95 25

N 8 8 8 8 8

169
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 11 (cont) Organ weights - group mean values relative (%) to brain weight (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group/ 

Sex Brain Heart Kidneys Liver

Lungs and 

Bronchi

Statisticstest Wi Wi Wi Wi Wi

1F Mean 60.711 83.5 87.7 371 137

SD 4.339 9.2 11.2 64 13

N 8 8 8 8 8

2F Mean 57.416 85.4 91.2 439 143

SD 3.493 6.3 14.0 74 14

N 8 8 8 8 7

3F Mean 55.358* 93.9 95.6 493** 158*

SD 4.127 13.0 23.5 89 24

N 8 8 8 8 8

170
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 12 Macropathology - group distribution of findings

Terminal sacrifice

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Number of animals affected

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

Tissue/Organ and Findings No. of animals 8 7 8 8 8 8

Number of animals within normal limits 7 7 5 7 5 5

Heart

 Adhesion(s) 0 0 1 0 0 0

 Pericaridial Fluid 0 0 1 0 0 0

Kidneys

 Cyst(s) 0 0 0 1 0 0

Lungs and Bronchi

 Dark area(s) 1 0 1 0 1 2

Lymph Node, Mandibular

 Dark 0 0 0 0 1 1

Rectum

 Dark area(s) 0 0 1 0 0 0

Stomach

 Dark area(s) 0 0 0 0 1 1

171
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

TABLE 12 (cont) Macropathology - group distribution of findings

Unscheduled sacrifice

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Number of animals affected

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

Tissue/Organ and Findings No. of animals 0 1 0 0 0 0

Colon

 Abnormal contents 0 1 0 0 0 0

Heart

 Thrombus 0 1 0 0 0 0

Rectum

 Prolapse 0 1 0 0 0 0

172
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

1M 1145 29 Normal Within normal limits P5-P15, 1-3, 19-

29

P5-P15, 1-2, 16, 

19-28

 

Skin Scab, Ears 7-18 7-15, 17-18 

Scab, Hindlimb - Right 4-8 3-7 

Scab, Tail 14-17 13-15, 17 

Ulceration, Tail 13 

Skin abrasion Dry, Head 6 

Dry, Ventral Surface P2-P4 P1-P4 

1146 29 Normal Within normal limits P5-P15, 1-29 P5-P15, 1-28 

Skin abrasion Dry, Ears P2-P4 P1-P4 

1147 29 Normal Within normal limits P5-P15, 1-29 P5-P15, 1-28 

Skin abrasion Dry, Ventral Surface P2-P4 P1-P4 

173
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

1M 1148 29 Normal Within normal limits P5-P15, 1-13, 15, 

17, 20, 24-29

P5-P15, 1-12, 14-

17, 20, 24-28

 

Discharge Aqueous, Red, Penis 16 

Viscous, Red, Penis 14 13 13

Exudate Red Exudate In Cage 16 

Skin Scab, Head 18-19 18-19 

Skin abrasion Dry, Ear - Left 21-23 21-23 

Dry, Ears P2-P4 P1-P4 

Dry, Ventral Surface P2-P4 P1-P4 

1149 29 Normal Within normal limits P2-P15, 1-29 P1-P15, 1-28 

1150 29 Normal Within normal limits P2-P11, P13-P15, 

1-19, 22-29

P1-P10, P13-P15, 

1-19, 21-28

 

Skin Bruise, HindLimb -Right-Slight P11 

Bruise, Hindlimb - Right P12 P12 

Scab, Hindlimb - Right 20-21 20 

1151 29 Normal Within normal limits P2-P15, 1-29 P1-P15, 1-28 

1152 29 Normal Within normal limits P2-P15, 1-15, 21-

29

P1-P15, 1-15, 21-

28

 

Skin abrasion Dry, Tail 16-20 16-20 

174
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

2M 2145 29 Normal Within normal limits P2-P15, 1-29 P1-P15, 1-28 

2146 29 Normal Within normal limits P2-P15, 1-29 P1-P15, 1-28 

2147 16 Normal Within normal limits P2-P15, 1-15 P1-P15, 1-15 

Discharge Aqueous, Red, Anus 16 16 

Reason for dispatch Prolapsed 16 

2149 29 Normal Within normal limits P2-P15, 1-16, 21-

29

P1-P15, 1-16, 21-

28

 

Skin abrasion Dry, Hindlimb - Right 17-20 17-20 

2150 29 Normal Within normal limits P2-P15, 1-6, 11-

29

P1-P15, 1-6, 11-

28

 

Skin abrasion Dry, Head 7-10 7-10 

2151 29 Normal Within normal limits P2-P15, 1-13, 16-

17, 21-29

P1-P15, 1-13, 16-

17, 21-27

 

Skin Scab, Dorsal surface 18-20 18-20 

Skin abrasion Dry, Head 14-15 14-15 

Dry, Upper Dorsal Surface 28 

2152 29 Normal Within normal limits P2-P15, 1-29 P1-P15, 1-28 

175
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

2M 2153 29 Normal Within normal limits P7-P15, 1-4, 6-29 P6-P15, 1-4, 6-28 

Skin Scab, Ear - Left P6 

Skin abrasion Dry, Ears P2-P5 P1-P5 

Dry, Upper Dorsal Surface 5 5 

3M 3145 29 Normal Within normal limits P2-P15, 1-4, 16, 

24, 28-29

P1-P15, 1-4, 28 

Skin Scab, Lower Dorsal Surface 7-15 7-15 

Scab, Upper Dorsal Surface 12-15, 26-27 12-15, 25-27 

Skin abrasion Dry, Dorsal surface 25-27 24-27 

Dry, Head 5-6 5-6 4

Dry, Lower Dorsal Surface 5-11 5-11 4

Dry, Upper Dorsal Surface 14, 17-23 16-23 

3146 29 Normal Within normal limits P2-P14, 2-4, 11-

13, 16-17, 21-29

P1-P13, 2-4, 10-

13, 16-17, 21-28

 

Skin Scab, Dorsal surface 7-10, 14-15 7-9, 14-15 

Scab, Head 18-20 18-20 

Skin abrasion Dry, Dorsal surface P15, 1, 7-10 P15, 1, 7-9 

Dry, Head 5-6 P14, 5-6 

Dry, Upper Dorsal Surface 5-6 5-6 

176
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

3M 3147 29 Normal Within normal limits P2-P12, P14, 11-

13, 28-29

P1-P11, P13-P14, 

11-13, 28

 

Skin Scab, Ear - Right P13 

Scab, Head 16-27 16-27 

Skin abrasion Dry, Ear - Left 2-6 1-6 4

Dry, Head P15, 1, 7-10, 14-

15

P15, 7-10, 14-15 

Dry, Lower Dorsal Surface 5-6 5-6 4

Dry, Upper Dorsal Surface 7-10, 14-15 7-10, 14-15 

Wet, Ear - Right P12 

3148 29 Normal Within normal limits P2-P14, 16-29 P1-P14, 16-28 

Skin Scab, Ear - Left 7-15 7-15 

Scab, Upper Dorsal Surface 12-15 12-15 

Skin abrasion Dry, Head P15, 1-6 P15, 1-6 

Dry, Upper Dorsal Surface 5-11 5-11 

3149 29 Normal Within normal limits P9-P15, 1-5, 7-19, 

22-29

P1, P3, P9-P15, 

1-5, 7-19, 22-28

 

Skin Scab, Ear - Left P6-P8 P6-P8 

Scab, Head 20-21 20-21 

Skin abrasion Dry, Ear - Left 6 6 

Dry, Ears P2-P5 P2, P4-P5 

177
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

3M 3150 29 Normal Within normal limits P2-P15, 1-29 P1-P15, 1-28 

3151 29 Normal Within normal limits P11-P15, 1-18, 

24-29

P10-P15, 1-18, 

24-28

 

Skin Scab, Ear - Left P6-P10 P6-P9 

Scab, Hindlimb - Right 19-23 19-23 

Skin abrasion Dry, Ears P2-P5 P1-P5 

3152 29 Normal Within normal limits P2-P15, 1-17, 24-

29

P1-P15, 1-17, 24-

28

 

Skin Scab, Ear - Left 22-23 21-23 

Ulceration, Ear - Left 18-21 18-20 

178
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

1F 1645 29 Normal Within normal limits P2, P6-P15, Ra1, 

1-3, 6-29

P1, P6-P15, Ra1, 

1-3, 6-28

 

Skin Scab, Head P5 P5 

Skin abrasion Dry, Head P3-P4 P2-P4 

Dry, Upper Dorsal Surface 4-5 4-5 

1646 29 Normal Within normal limits P2-P15, Ra1, 1-

29

P1-P15, Ra1, 1-

28

 

1647 29 Normal Within normal limits P2-P15, Ra1, 1-

16, 21-29

P1-P15, Ra1, 1-

16, 21-28

 

Skin abrasion Dry, Hindlimb - Right 17-20 17-20 

1648 29 Normal Within normal limits P2-P15, Ra1, 1-

15, 18-29

P1-P15, Ra1, 1-

15, 17-28

 

Skin abrasion Dry, Head 16-17 16 

1649 29 Normal Within normal limits P5-P15, Ra1, 1-3, 

7-29

P5-P15, Ra1, 1-3, 

7-9, 11-28

 

Skin abrasion Dry, Ears P2-P4 P1-P4 

Dry, Nose 10 

Dry, Upper Dorsal Surface 4-6 4-6 

1650 29 Normal Within normal limits P2-P15, Ra1, 1-

29

P1-P15, Ra1, 1-

28

 

179
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

1F 1651 29 Normal Within normal limits P2-P15, Ra1, 1-3, 

6-10, 12-17, 23-

29

P1-P15, Ra1, 1-3, 

6-9, 12-17, 23-28

 

Skin Scab, Ear - Left 18-22 18-22 

Skin abrasion Dry, Head 11 10-11 

Dry, Hindlimb - Right 4-5 4-5 

1652 29 Normal Within normal limits P2-P15, Ra1, 1-

29

P1-P15, Ra1, 1-

28

 

2F 2645 29 Normal Within normal limits P5-P15, Ra1, 1-

16, 23-29

P5-P15, Ra1, 1-

16, 23-28

 

Skin abrasion Dry, Ear - Left 17-22 17-22 

Dry, Ears P2-P4 P1-P4 

2646 29 Normal Within normal limits P7-P15, Ra1, 1-

18, 23-29

P7-P15, Ra1, 1-

18, 23-28

 

Skin abrasion Dry, Head 19-22 19-22 

Dry, Ventral Surface P2-P6 P1-P6 

2647 29 Normal Within normal limits P2-P15, Ra1, 1-

24, 28-29

P1-P15, Ra1, 1-

24, 28

 

Skin abrasion Dry, Dorsal surface 25-27 25-27 

180
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

2F 2648 29 Normal Within normal limits P2-P15, Ra1, 1-5, 

7-29

P1-P15, Ra1, 1-5, 

7-28

 

Skin abrasion Dry, Lower Dorsal Surface 6 6 

2649 29 Normal Within normal limits P2-P15, Ra1, 1-4, 

7-10, 12-18, 23-

29

P1-P15, Ra1, 1-4, 

7-9, 12-18, 23-28

 

Skin Scab, Hindlimb - Right 6 6 

Skin abrasion Dry, Dorsal surface 21-22 20-22 

Dry, Head 19-22 19-22 

Dry, Hindlimb - Right 5 5 

Dry, Upper Ventral Surface 10 

Skin color Abnormal color, Red, Upper Dorsal Surface 11 10-11 

2650 29 Normal Within normal limits P2-P15, Ra1, 1-

29

P1-P15, Ra1, 1-

28

 

2651 29 Normal Within normal limits P2-P15, Ra1, 1-

29

P1-P15, Ra1, 1-

28

 

2652 29 Normal Within normal limits P2, P5-P15, Ra1, 

1-29

P1, P5-P15, Ra1, 

1-28

 

Skin abrasion Dry, Upper Dorsal Surface P3-P4 P2-P4 

181
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

3F 3645 29 Normal Within normal limits P2-P15, Ra1, 1-5, 

7-17, 21-29

P1-P15, Ra1, 1-5, 

7-17, 21-28

 

Skin Scab, Hindlimb - Right 18-20 18-20 

Skin abrasion Dry, Ear - Right 6 6 

3646 29 Normal Within normal limits P2-P15, Ra1, 1-3, 

6-9, 15-29

P1-P15, Ra1, 1-3, 

6-9, 15-28

 

Skin Scab, Lower Dorsal Surface 10-14 10-14 

Skin abrasion Dry, Head 4-5 4-5 

3647 29 Normal Within normal limits P2-P15, Ra1, 1-3, 

6-17, 23-29

P1-P15, Ra1, 1-3, 

6-17, 23-28

 

Skin Scab, Head 18-22 18-22 

Skin abrasion Dry, Head 4-5 4-5 

3648 29 Normal Within normal limits P2-P15, Ra1, 1-

29

P1-P15, Ra1, 1-

28

 

3649 29 Normal Within normal limits P2-P15, Ra1, 1-

29

P1-P15, Ra1, 1-

28

 

3650 29 Normal Within normal limits P2-P15, Ra1, 1-3, 

6-29

P1-P15, Ra1, 1-3, 

6-28

 

Skin abrasion Dry, Head 4-5 4-5 

Dry, Upper Dorsal Surface 5 5 

182
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 1 (cont) Daily observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation AM PM US PM

3F 3651 29 Normal Within normal limits P2-P15, Ra1, 1-

29

P1-P15, Ra1, 1-

28

 

3652 29 Normal Within normal limits P2-P15, Ra1, 1-

29

P1-P15, Ra1, 1-

28

 

183
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 2 Dose observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation 2 Hours 4 Hours US1 US2

1M 1145 29 Normal Within normal limits 1-28 1-28 

1146 29 Normal Within normal limits 1-28 1-28 

1147 29 Normal Within normal limits 1-28 1-28 

1148 29 Normal Within normal limits 1-13, 15-28 1-13, 15-28 

1149 29 Normal Within normal limits 1-28 1-28 

1150 29 Normal Within normal limits 1-28 1-28 

1151 29 Normal Within normal limits 1-28 1-28 

1152 29 Normal Within normal limits 1-28 1-28 

2M 2145 29 Normal Within normal limits 1-28 1-28 

2146 29 Normal Within normal limits 1-28 1-28 

2147 16 Normal Within normal limits 1-15 1-15 

184
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 2 (cont) Dose observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation 2 Hours 4 Hours US1 US2

2M 2149 29 Normal Within normal limits 1-28 1-28 

2150 29 Normal Within normal limits 1-28 1-28 

2151 29 Normal Within normal limits 1-28 1-28 

2152 29 Normal Within normal limits 1-28 1-28 

2153 29 Normal Within normal limits 2-28 2-28 

3M 3145 29 Normal Within normal limits 1-28 1-28 

3146 29 Normal Within normal limits 1-28 1-28 

3147 29 Normal Within normal limits 1-28 1-23, 25-28 

Muscle reaction Shivering 24 

3148 29 Normal Within normal limits 1-28 1-28 

3149 29 Normal Within normal limits 1-28 1-28 

3150 29 Normal Within normal limits 1-28 1-28 

185
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 2 (cont) Dose observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation 2 Hours 4 Hours US1 US2

3M 3151 29 Normal Within normal limits 1-28 1-28 

3152 29 Normal Within normal limits 1-28 1-28 

186
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 2 (cont) Dose observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation 2 Hours 4 Hours US1 US2

1F 1645 29 Normal Within normal limits 1-28 1-28 

1646 29 Normal Within normal limits 1-28 1-28 

1647 29 Normal Within normal limits 1-28 1-28 

1648 29 Normal Within normal limits 1-28 1-28 

1649 29 Normal Within normal limits 1-28 1-28 

1650 29 Normal Within normal limits 1-28 1-28 

1651 29 Normal Within normal limits 1-28 1-28 

1652 29 Normal Within normal limits 1-28 1-28 

2F 2645 29 Normal Within normal limits 1-28 1-28 

2646 29 Normal Within normal limits 1-28 1-28 

2647 29 Normal Within normal limits 1-28 1-28 

187
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 2 (cont) Dose observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation 2 Hours 4 Hours US1 US2

2F 2648 29 Normal Within normal limits 1-28 1-28 

General condition Laceration, Tail 4

2649 29 Normal Within normal limits 1-28 1-28 

2650 29 Normal Within normal limits 1-28 1-28 

2651 29 Normal Within normal limits 1-28 1-28 

2652 29 Normal Within normal limits 1-28 1-28 

3F 3645 29 Normal Within normal limits 1-28 1-28 

3646 29 Normal Within normal limits 1-28 1-28 

3647 29 Normal Within normal limits 1-28 1-28 

3648 29 Normal Within normal limits 1-28 1-28 

188
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 2 (cont) Dose observations - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of Day(s) observed

/Sex Number Death Category Observation 2 Hours 4 Hours US1 US2

3F 3649 29 Normal Within normal limits 1, 3-28 1, 3-28 

General condition Laceration, Tail 2 2 

Laceration, Tail, red 

exudate present

 1 

3650 29 Normal Within normal limits 1-28 1-28 

3651 29 Normal Within normal limits 1-28 1-28 

Breathing Coughing 11 

3652 29 Normal Within normal limits 1-28 1-28 

189
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 3 Clinical signs - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week of

/Sex Number Death Phase Category Observation Week(s) observed

1M 1145S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 4

Skin Scab, Ears 1-3

Scab, Hindlimb - Right 1-3

1146S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1147S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1148S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1, 3-4

Discharge Viscous, Red, Penis 2

1149S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1150S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1151S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

S1 - Scheduled Euthanasia 1

190
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 3 (cont) Clinical signs - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week of

/Sex Number Death Phase Category Observation Week(s) observed

1M 1152S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

2M 2145S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

2146S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

2147WE 3 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-2

2149S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

2150S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 2-4

Skin abrasion Dry, Head 1

2151S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1, 4

Skin abrasion Dry, Ear - Left 3

Dry, Head 2-3

S1 - Scheduled Euthanasia 1, WE - Euthanized for welfare reasons

191
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 3 (cont) Clinical signs - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week of

/Sex Number Death Phase Category Observation Week(s) observed

2M 2152S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

2153S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

3M 3145S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 4

Skin Scab, Lower Dorsal Surface 1-3

Skin abrasion Dry, Lower Dorsal Surface 1-3

3146S1 5 Pre-Treatment Normal Within normal limits P2

Skin abrasion Dry, Dorsal surface P3

Treatment Normal Within normal limits 4

Skin Scab, Dorsal surface 1-3

Skin abrasion Dry, Dorsal surface 1-3

3147S1 5 Pre-Treatment Normal Within normal limits P2

Skin abrasion Dry, Head P3

Treatment Normal Within normal limits 4

Skin abrasion Dry, Ear - Left 3

Dry, Head 1-3

Dry, Upper Dorsal Surface 1-3

S1 - Scheduled Euthanasia 1

192
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 3 (cont) Clinical signs - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week of

/Sex Number Death Phase Category Observation Week(s) observed

3M 3148S1 5 Pre-Treatment Normal Within normal limits P2

Skin abrasion Dry, Head P3

Treatment Normal Within normal limits 4

Skin Scab, Ear - Left 1-3

Scab, Upper Dorsal Surface 1-3

3149S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

3150S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

3151S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

3152S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-2, 4

Skin Ulceration, Ear - Left 3

S1 - Scheduled Euthanasia 1

193
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 3 (cont) Clinical signs - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week of

/Sex Number Death Phase Category Observation Week(s) observed

1F 1645S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1646S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1647S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1648S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1649S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1650S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1651S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

1652S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

S1 - Scheduled Euthanasia 1

194
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 3 (cont) Clinical signs - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week of

/Sex Number Death Phase Category Observation Week(s) observed

2F 2645S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-2, 4

Skin abrasion Dry, Ear - Left 3

2646S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-2, 4

Skin abrasion Dry, Ear - Left 3

2647S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

2648S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

2649S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

2650S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

2651S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

S1 - Scheduled Euthanasia 1

195
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 3 (cont) Clinical signs - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week of

/Sex Number Death Phase Category Observation Week(s) observed

2F 2652S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

3F 3645S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

3646S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1, 3-4

Skin Scab, Lower Dorsal Surface 2

3647S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-2, 4

Skin abrasion Dry, Head 3

3648S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

3649S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

3650S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

S1 - Scheduled Euthanasia 1

196
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 3 (cont) Clinical signs - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week of

/Sex Number Death Phase Category Observation Week(s) observed

3F 3651S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

3652S1 5 Pre-Treatment Normal Within normal limits P2-P3

Treatment Normal Within normal limits 1-4

S1 - Scheduled Euthanasia 1

197
Sponsor refe

Huntingdo

APPENDIX 4 Body weights - individual values (kg)  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Week Week Change Chan

/Sex Number P1 P2 P3 1 2 3 4 P3-1 P3-2 

1M 1145S1 7.8 8.4 8.7 9.5 10.3 10.8 11.4 0.8 1.6 

 1146S1 6.5 6.8 7.1 7.9 8.3 9.3 9.5 0.8 1.2  1147S1 6.3 7.2 7.5 8.4 9.3 10.8 11.8 0.9 1.8  1148S1 6.8 7.7 8.3 9.4 10.0 11.1 11.6 1.1 1.7  1149S1 6.5 6.8 7.5 8.6 9.8 11.0 11.5 1.1 2.3  1150S1 8.1 8.2 8.9 9.9 10.8 11.9 12.5 1.0 1.9  1151S1 5.4 5.9 6.3 7.0 7.6 8.3 8.8 0.7 1.3  1152S1 6.7 6.7 7.0 7.8 8.4 9.4 9.8 0.8 1.4 

2M 2145S1 5.8 6.0 6.5 7.3 7.9 9.1 9.8 0.8 1.4 

 2146S1 5.4 6.1 6.1 6.8 7.4 8.2 8.9 0.7 1.3  2147WE 7.2 7.7 8.4 9.7 10.5 1.3 2.1  2149S1 5.6 5.6 6.1 6.7 7.3 8.2 8.7 0.6 1.2  2150S1 6.6 7.3 8.0 8.9 9.7 10.8 11.3 0.9 1.7  2151S1 6.9 7.3 7.9 8.5 9.3 10.3 10.9 0.6 1.4  2152S1 6.0 6.6 7.3 8.1 9.2 10.5 11.1 0.8 1.9  2153S1 7.0 7.3 8.1 8.5 9.3 10.3 10.9 0.4 1.2 

S1 - Scheduled Euthanasia 1, WE - Euthanized for welfare reasons 

198
Sponsor refe

Huntingdo

APPENDIX 4 (cont) Body weights - individual values (kg)  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Week Week Change Chan

/Sex Number P1 P2 P3 1 2 3 4 P3-1 P3-2 

3M 3145S1 4.9 5.5 6.0 6.7 7.3 8.0 8.2 0.7 1.3 

 3146S1 5.1 5.7 6.4 6.9 7.3 7.8 8.1 0.5 0.9  3147S1 6.5 7.4 8.2 9.3 10.2 10.7 11.2 1.1 2.0  3148S1 6.1 6.7 7.2 8.2 9.5 11.4 11.8 1.0 2.3  3149S1 5.7 5.9 6.1 6.5 7.2 8.1 8.2 0.4 1.1  3150S1 6.2 6.7 7.7 8.3 8.9 9.8 10.0 0.6 1.2  3151S1 7.5 8.0 8.7 9.6 10.6 11.9 12.2 0.9 1.9  3152S1 5.5 5.9 6.5 7.1 8.0 8.8 9.1 0.6 1.5 

S1 - Scheduled Euthanasia 1 

199
Sponsor refe

Huntingdo

APPENDIX 4 (cont) Body weights - individual values (kg)  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Week Week Change Chan

/Sex Number P1 P2 P3 1 2 3 4 P3-1 P3-2 

1F 1645S1 7.0 7.9 8.6 9.8 10.4 11.5 12.3 1.2 1.8 

 1646S1 7.1 7.4 8.3 9.5 10.3 11.2 11.8 1.2 2.0  1647S1 7.3 8.0 8.7 10.1 10.8 11.8 12.4 1.4 2.1  1648S1 6.5 6.8 7.2 8.0 8.4 9.4 10.0 0.8 1.2  1649S1 7.0 8.0 9.0 10.5 11.4 12.8 13.5 1.5 2.4  1650S1 6.1 6.8 7.5 8.5 9.3 10.3 11.1 1.0 1.8  1651S1 6.6 7.0 7.3 8.4 9.2 10.2 10.7 1.1 1.9  1652S1 7.1 7.7 8.2 9.1 9.8 10.6 11.5 0.9 1.6 

2F 2645S1 5.7 6.2 6.9 7.9 8.2 9.4 9.9 1.0 1.3 

 2646S1 8.2 8.9 9.7 10.8 11.9 13.6 14.3 1.1 2.2  2647S1 5.7 5.9 6.5 7.4 8.1 9.2 9.7 0.9 1.6  2648S1 6.1 6.8 7.4 8.5 9.2 10.3 11.4 1.1 1.8  2649S1 7.8 8.4 9.2 10.3 10.9 12.4 12.8 1.1 1.7  2650S1 6.4 6.5 6.9 8.0 8.4 9.9 10.6 1.1 1.5  2651S1 6.4 7.3 7.8 9.4 10.4 11.7 12.7 1.6 2.6  2652S1 5.2 5.7 6.3 7.2 8.1 9.3 10.3 0.9 1.8 

S1 - Scheduled Euthanasia 1 

200
Sponsor refe

Huntingdo

APPENDIX 4 (cont) Body weights - individual values (kg)  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Week Week Change Chan

/Sex Number P1 P2 P3 1 2 3 4 P3-1 P3-2 

3F 3645S1 7.1 7.2 8.0 9.4 10.2 12.0 12.9 1.4 2.2 

 3646S1 7.2 8.0 8.5 9.6 10.5 11.8 12.3 1.1 2.0  3647S1 5.7 6.1 6.5 7.3 8.0 9.0 9.4 0.8 1.5  3648S1 7.0 7.9 8.6 9.6 10.5 11.7 12.5 1.0 1.9  3649S1 6.2 6.4 7.1 8.2 9.0 10.2 11.3 1.1 1.9  3650S1 6.4 6.8 7.1 7.9 8.5 9.2 9.6 0.8 1.4  3651S1 7.1 8.0 8.9 10.0 11.2 12.7 13.6 1.1 2.3  3652S1 6.9 7.3 7.9 9.1 10.1 11.2 12.0 1.2 2.2 

S1 - Scheduled Euthanasia 1 

201
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 5 Body weight change - individual values (kg)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week Week

/Sex Number P1-P2 P2-P3 P3-1 1-2 2-3 3-4

1M 1145S1 0.6 0.3 0.8 0.8 0.5 0.6

1146S1 0.3 0.3 0.8 0.4 1.0 0.2

1147S1 0.9 0.3 0.9 0.9 1.5 1.0

1148S1 0.9 0.6 1.1 0.6 1.1 0.5

1149S1 0.3 0.7 1.1 1.2 1.2 0.5

1150S1 0.1 0.7 1.0 0.9 1.1 0.6

1151S1 0.5 0.4 0.7 0.6 0.7 0.5

1152S1 0.0 0.3 0.8 0.6 1.0 0.4

2M 2145S1 0.2 0.5 0.8 0.6 1.2 0.7

2146S1 0.7 0.0 0.7 0.6 0.8 0.7

2147WE 0.5 0.7 1.3 0.8

2149S1 0.0 0.5 0.6 0.6 0.9 0.5

2150S1 0.7 0.7 0.9 0.8 1.1 0.5

2151S1 0.4 0.6 0.6 0.8 1.0 0.6

2152S1 0.6 0.7 0.8 1.1 1.3 0.6

2153S1 0.3 0.8 0.4 0.8 1.0 0.6

S1 - Scheduled Euthanasia 1, WE - Euthanized for welfare reasons

202
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 5 (cont) Body weight change - individual values (kg)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week Week

/Sex Number P1-P2 P2-P3 P3-1 1-2 2-3 3-4

3M 3145S1 0.6 0.5 0.7 0.6 0.7 0.2

3146S1 0.6 0.7 0.5 0.4 0.5 0.3

3147S1 0.9 0.8 1.1 0.9 0.5 0.5

3148S1 0.6 0.5 1.0 1.3 1.9 0.4

3149S1 0.2 0.2 0.4 0.7 0.9 0.1

3150S1 0.5 1.0 0.6 0.6 0.9 0.2

3151S1 0.5 0.7 0.9 1.0 1.3 0.3

3152S1 0.4 0.6 0.6 0.9 0.8 0.3

S1 - Scheduled Euthanasia 1

203
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 5 (cont) Body weight change - individual values (kg)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week Week

/Sex Number P1-P2 P2-P3 P3-1 1-2 2-3 3-4

1F 1645S1 0.9 0.7 1.2 0.6 1.1 0.8

1646S1 0.3 0.9 1.2 0.8 0.9 0.6

1647S1 0.7 0.7 1.4 0.7 1.0 0.6

1648S1 0.3 0.4 0.8 0.4 1.0 0.6

1649S1 1.0 1.0 1.5 0.9 1.4 0.7

1650S1 0.7 0.7 1.0 0.8 1.0 0.8

1651S1 0.4 0.3 1.1 0.8 1.0 0.5

1652S1 0.6 0.5 0.9 0.7 0.8 0.9

2F 2645S1 0.5 0.7 1.0 0.3 1.2 0.5

2646S1 0.7 0.8 1.1 1.1 1.7 0.7

2647S1 0.2 0.6 0.9 0.7 1.1 0.5

2648S1 0.7 0.6 1.1 0.7 1.1 1.1

2649S1 0.6 0.8 1.1 0.6 1.5 0.4

2650S1 0.1 0.4 1.1 0.4 1.5 0.7

2651S1 0.9 0.5 1.6 1.0 1.3 1.0

2652S1 0.5 0.6 0.9 0.9 1.2 1.0

S1 - Scheduled Euthanasia 1

204
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 5 (cont) Body weight change - individual values (kg)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Week Week

/Sex Number P1-P2 P2-P3 P3-1 1-2 2-3 3-4

3F 3645S1 0.1 0.8 1.4 0.8 1.8 0.9

3646S1 0.8 0.5 1.1 0.9 1.3 0.5

3647S1 0.4 0.4 0.8 0.7 1.0 0.4

3648S1 0.9 0.7 1.0 0.9 1.2 0.8

3649S1 0.2 0.7 1.1 0.8 1.2 1.1

3650S1 0.4 0.3 0.8 0.6 0.7 0.4

3651S1 0.9 0.9 1.1 1.2 1.5 0.9

3652S1 0.4 0.6 1.2 1.0 1.1 0.8

S1 - Scheduled Euthanasia 1

205
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

1M 1145S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1146S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1147S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1148S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1149S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

S1 - Scheduled Euthanasia 1

206
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

1M 1149S1 29 Treatment Visual Food 

Consumption

4: All food consumed 1-28

1150S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1151S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1152S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2M 2145S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

S1 - Scheduled Euthanasia 1

207
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

2M 2146S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2147WE 16 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-15

2149S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2150S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2151S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

S1 - Scheduled Euthanasia 1, WE - Euthanized for welfare reasons

208
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

2M 2151S1 29 Treatment Visual Food 

Consumption

4: All food consumed 1-28

2152S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2153S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3M 3145S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3146S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

S1 - Scheduled Euthanasia 1

209
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

3M 3147S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3148S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3149S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3150S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3151S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

S1 - Scheduled Euthanasia 1

210
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

3M 3151S1 29 Treatment Visual Food 

Consumption

4: All food consumed 1-28

3152S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

Treatment Visual Food 

Consumption

4: All food consumed 1-28

S1 - Scheduled Euthanasia 1

211
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

1F 1645S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1646S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1647S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1648S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

S1 - Scheduled Euthanasia 1

212
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

1F 1648S1 29 RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1649S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1650S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

1651S1 29 Pre-Treatment Visual Food

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

S1 - Scheduled Euthanasia 1

213
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

1F 1651S1 29 Treatment Visual Food 

Consumption

4: All food consumed 1-28

1652S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2F 2645S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2646S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

S1 - Scheduled Euthanasia 1

214
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

2F 2647S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2648S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2649S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2650S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

S1 - Scheduled Euthanasia 1

215
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

2F 2650S1 29 RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2651S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

2652S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3F 3645S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

S1 - Scheduled Euthanasia 1

216
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

3F 3645S1 29 Treatment Visual Food 

Consumption

4: All food consumed 1-28

3646S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3647S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3648S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

S1 - Scheduled Euthanasia 1

217
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

3F 3649S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3650S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3651S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

3652S1 29 Pre-Treatment Visual Food 

Consumption

4: All food consumed P8-P15

S1 - Scheduled Euthanasia 1

218
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 6 (cont) Visual food consumption - individual observations

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group Animal Day of

/Sex Number Death Phase Category Observation Day(s) observed

3F 3652S1 29 RandomizationVisual Food 

Consumption

4: All food consumed Ra1

Treatment Visual Food 

Consumption

4: All food consumed 1-28

S1 - Scheduled Euthanasia 1

219
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

1145 Pretest 1 11.2 33.4 6.77 218.5 991 49.3 16.5 33.4 18.6 16.87

Pretest 2 12.6 37.7 7.52 247.7 939 50.1 16.7 33.3 18.3 10.95

Term Day 21 13.4 39.1 8.05 186.9 739 48.6 16.7 34.3 17.3 9.62

Term Day 28 14.4 42.3 8.60 162.7 677 49.2 16.7 34.0 16.8 8.85

1146 Pretest 1 14.0 44.3 8.26 263.3 727 53.7 17.0 31.7 16.9 15.97

Pretest 2 13.9 40.6 7.93 152.8 664 51.3 17.5 34.2 16.5 14.35

Term Day 21 13.4 38.8 7.78 176.0 584 49.9 17.2 34.5 16.6 20.32

Term Day 28 13.4 40.8 7.83 149.1 756 52.1 17.2 32.9 19.2 10.11

1147 Pretest 1 13.0 41.3 9.05 345.1 328 45.6 14.3 31.4 17.1 13.92

Pretest 2 12.3 38.1 8.45 280.3 355 45.0 14.5 32.3 16.8 10.49

Term Day 21 12.9 39.1 8.68 283.2 371 45.0 14.8 32.9 17.4 9.78

Term Day 28 14.3 44.8 9.65 290.1 249 46.4 14.8 31.9 17.4 10.69

1148 Pretest 1 12.2 38.2 8.12 258.5 756 47.1 15.0 31.9 18.8 11.27

Pretest 2 12.9 39.8 8.41 209.8 496 47.4 15.3 32.3 18.3 8.59

Term Day 21 12.9 39.8 7.74 361.3 687 51.4 16.7 32.5 21.6 6.49

Term Day 28 15.5 47.9 9.48 136.4 370 50.5 16.3 32.3 18.7 7.66

1149 Pretest 1 12.0 36.7 6.58 240.5 530 55.8 18.3 32.8 16.0 13.50

Pretest 2 12.4 36.6 6.77 148.0 535 54.1 18.3 33.8 15.8 11.14

Term Day 21 12.0 35.8 6.69 79.8 438 53.5 17.9 33.6 15.5 9.52

Term Day 28 13.2 39.0 7.29 113.4 456 53.5 18.1 33.9 15.6 10.30

220
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

1145 Pretest 1 11.27 5.05 0.19 0.10 0.02 0.23

Pretest 2 5.22 5.25 0.21 0.11 0.02 0.14

Term Day 21 5.22 3.97 0.17 0.10 0.01 0.14

Term Day 28 4.31 4.12 0.22 0.14 0.02 0.04

1146 Pretest 1 5.46 9.88 0.24 0.14 0.03 0.22

Pretest 2 5.10 8.59 0.25 0.14 0.02 0.25

Term Day 21 10.86 8.80 0.23 0.06 0.17 0.20

Term Day 28 3.72 5.85 0.29 0.20 0.02 0.03

1147 Pretest 1 4.50 8.84 0.40 0.11 0.02 0.04 NCLP

Pretest 2 3.38 6.74 0.15 0.07 0.02 0.14

Term Day 21 3.48 5.90 0.17 0.10 0.02 0.12

Term Day 28 2.97 7.15 0.27 0.24 0.02 0.03 CLSL

1148 Pretest 1 5.50 5.15 0.31 0.12 0.02 0.18

Pretest 2 3.30 4.67 0.25 0.18 0.01 0.17

Term Day 21 2.99 3.13 0.20 0.06 0.01 0.10

Term Day 28 3.00 4.17 0.25 0.19 0.01 0.04

1149 Pretest 1 3.80 8.99 0.23 0.15 0.02 0.31

Pretest 2 3.81 6.78 0.18 0.12 0.01 0.24

Term Day 21 1.95 6.93 0.21 0.16 0.01 0.25

Term Day 28 2.70 6.97 0.35 0.20 0.02 0.06

221
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

1150 Pretest 1 11.6 35.9 7.27 260.4 535 49.4 15.9 32.3 17.4 8.19

Pretest 2 12.7 38.7 7.98 191.4 539 48.5 15.9 32.9 17.3 9.91

Term Day 21 12.9 40.0 8.16 204.7 463 49.0 15.8 32.3 17.0 8.94

Term Day 28 13.0 39.1 8.11 124.4 473 48.2 16.0 33.3 16.9 7.41

1151 Pretest 1 12.1 37.9 8.82 278.3 595 42.9 13.7 32.0 17.5 15.49

Pretest 2 11.5 35.3 8.58 187.6 608 41.2 13.4 32.5 17.3 13.16

Term Day 21 11.8 35.8 8.68 178.3 471 41.3 13.6 33.0 17.1 11.52

Term Day 28 12.0 36.4 8.52 179.7 537 42.7 14.0 32.9 18.7 13.13

1152 Pretest 1 12.8 40.1 6.63 294.8 557 60.4 19.2 31.8 16.9 13.33

Pretest 2 13.9 42.7 7.12 200.5 496 60.0 19.6 32.6 16.2 13.50

Term Day 21 14.6 44.7 7.60 158.0 298 58.8 19.2 32.7 15.3 10.76

Term Day 28 14.5 43.4 7.49 160.2 451 58.0 19.4 33.4 15.9 9.90

Group 2

2145 Pretest 1 11.2 35.9 7.29 185.2 603 49.2 15.4 31.2 19.1 12.37

Pretest 2 11.5 36.5 7.61 161.5 556 48.0 15.1 31.4 18.5 8.96

Term Day 21 11.8 36.9 7.96 156.4 493 46.4 14.8 31.9 18.4 8.68

Term Day 28 13.2 40.8 8.66 178.1 494 47.1 15.3 32.5 19.6 9.47

222
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

1150 Pretest 1 2.69 5.07 0.17 0.05 0.01 0.19

Pretest 2 5.32 4.21 0.15 0.05 0.01 0.17

Term Day 21 1.90 6.41 0.22 0.17 0.02 0.22

Term Day 28 2.15 4.77 0.26 0.18 0.01 0.04

1151 Pretest 1 5.94 8.87 0.27 0.11 0.02 0.28

Pretest 2 7.11 5.56 0.19 0.09 0.00 0.21

Term Day 21 4.57 6.29 0.36 0.25 0.00 0.04

Term Day 28 5.36 7.00 0.25 0.42 0.01 0.09

1152 Pretest 1 4.55 8.02 0.50 0.20 0.03 0.03

Pretest 2 6.91 5.90 0.43 0.08 0.01 0.17

Term Day 21 4.26 5.75 0.35 0.25 0.01 0.13 CLSE

Term Day 28 4.13 5.14 0.39 0.18 0.01 0.04

Group 2

2145 Pretest 1 4.61 7.14 0.17 0.15 0.02 0.28

Pretest 2 3.44 5.11 0.12 0.09 0.01 0.19

Term Day 21 3.03 5.10 0.22 0.15 0.01 0.18

Term Day 28 3.71 5.22 0.28 0.20 0.00 0.05

223
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 2

2146 Pretest 1 11.3 36.8 8.23 236.2 472 44.7 13.7 30.8 18.8 12.44

Pretest 2 11.0 34.3 8.02 140.5 450 42.7 13.8 32.2 18.7 9.49

Term Day 21 11.3 35.4 8.56 127.4 363 41.4 13.1 31.8 17.6 8.96

Term Day 28 12.6 39.4 9.32 170.3 355 42.3 13.5 32.0 19.0 8.17

2147 Pretest 1 11.7 35.7 6.79 237.9 550 52.6 17.3 32.8 17.4 10.61

Pretest 2 11.4 34.1 6.66 128.3 638 51.2 17.2 33.6 16.5 9.07

Term Day 21 NS NS NS NS NS NS NS NS NS NS

Term Day 28 NS NS NS NS NS NS NS NS NS NS

2149 Pretest 1 10.7 34.4 7.36 189.8 581 46.7 14.6 31.3 19.1 11.25

Pretest 2 10.2 32.4 7.18 173.8 456 45.1 14.3 31.6 18.5 8.97

Term Day 21 10.7 33.5 7.48 161.6 462 44.8 14.3 31.9 18.5 6.87

Term Day 28 11.9 37.3 8.31 187.9 384 44.9 14.4 32.0 19.1 6.44

2150 Pretest 1 11.1 35.0 6.85 321.6 691 51.0 16.2 31.8 18.8 9.29

Pretest 2 11.6 35.9 6.97 331.6 764 51.6 16.6 32.3 18.9 8.03

Term Day 21 12.1 35.8 7.31 86.4 589 48.9 16.5 33.7 15.9 5.63

Term Day 28 13.0 38.8 7.80 135.0 631 49.7 16.7 33.6 16.3 6.11

2151 Pretest 1 12.6 38.9 8.04 242.2 644 48.4 15.7 32.4 18.4 11.74

Pretest 2 12.9 39.5 8.27 190.6 380 47.7 15.6 32.7 18.2 9.11

Term Day 21 13.2 38.5 8.26 58.8 594 46.6 15.9 34.2 17.0 8.06

Term Day 28 12.9 37.1 7.89 61.1 604 47.0 16.4 34.8 17.2 7.72

224
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 2

2146 Pretest 1 4.72 6.95 0.28 0.23 0.02 0.25

Pretest 2 4.02 5.14 0.10 0.08 0.00 0.14

Term Day 21 3.95 4.40 0.20 0.21 0.00 0.20

Term Day 28 3.39 4.38 0.21 0.15 0.01 0.04

2147 Pretest 1 3.04 6.71 0.24 0.27 0.02 0.33

Pretest 2 3.20 5.36 0.17 0.13 0.02 0.19

Term Day 21 NS NS NS NS NS NS

Term Day 28 NS NS NS NS NS NS

2149 Pretest 1 5.29 5.29 0.23 0.12 0.01 0.31

Pretest 2 4.14 4.19 0.29 0.10 0.01 0.24

Term Day 21 1.99 3.96 0.40 0.20 0.01 0.32

Term Day 28 1.72 4.09 0.38 0.19 0.00 0.05

2150 Pretest 1 3.77 5.08 0.13 0.09 0.00 0.21

Pretest 2 2.82 4.80 0.14 0.09 0.01 0.16

Term Day 21 1.52 3.73 0.16 0.08 0.00 0.13

Term Day 28 1.78 3.97 0.19 0.12 0.00 0.04

2151 Pretest 1 5.97 5.19 0.39 0.13 0.01 0.04

Pretest 2 4.00 4.39 0.21 0.34 0.01 0.16

Term Day 21 3.17 4.25 0.24 0.23 0.01 0.14

Term Day 28 3.06 4.10 0.33 0.18 0.01 0.03

225
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 2

2152 Pretest 1 10.1 31.6 6.25 318.2 871 50.6 16.1 31.9 19.4 10.99

Pretest 2 10.8 33.7 6.61 320.4 866 51.0 16.3 32.1 18.7 9.72

Term Day 21 11.9 36.4 7.47 279.1 602 48.7 15.9 32.6 17.6 10.02

Term Day 28 12.1 36.5 7.50 205.8 641 48.7 16.1 33.1 17.2 8.43

2153 Pretest 1 13.7 41.6 8.40 264.9 673 49.6 16.3 32.8 17.3 17.10

Pretest 2 13.8 41.9 8.53 182.2 600 49.2 16.2 32.9 16.9 13.60

Term Day 21 14.1 41.7 8.90 140.2 500 46.9 15.9 33.8 16.1 11.44

Term Day 28 14.4 43.0 8.99 201.2 522 47.8 16.1 33.6 16.9 12.50

Group 3

3145 Pretest 1 12.6 38.6 6.47 320.0 1077 59.6 19.4 32.6 18.5 9.58

Pretest 2 13.3 41.7 6.81 345.7 900 61.3 19.6 31.9 17.8 10.49

Term Day 21 13.7 40.5 6.99 136.4 755 58.0 19.6 33.9 15.8 9.21

Term Day 28 13.4 39.7 6.89 67.5 781 57.6 19.4 33.7 15.4 10.72

3146 Pretest 1 11.5 36.4 7.91 271.3 494 46.0 14.5 31.5 17.9 13.16

Pretest 2 11.5 36.5 7.89 226.0 517 46.3 14.6 31.5 17.2 14.53

Term Day 21 13.0 38.7 9.03 118.4 250 42.9 14.4 33.6 15.7 11.44

Term Day 28 12.4 38.4 8.69 173.9 466 44.2 14.2 32.2 16.5 9.40

226
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 2

2152 Pretest 1 4.64 5.71 0.37 0.12 0.01 0.13

Pretest 2 3.71 5.53 0.24 0.07 0.01 0.15

Term Day 21 4.01 5.41 0.29 0.15 0.01 0.14

Term Day 28 3.20 4.76 0.28 0.15 0.01 0.03

2153 Pretest 1 5.27 10.95 0.29 0.22 0.03 0.34

Pretest 2 3.88 9.09 0.21 0.15 0.02 0.25

Term Day 21 3.22 7.50 0.28 0.23 0.01 0.20

Term Day 28 2.96 8.88 0.34 0.23 0.01 0.07

Group 3

3145 Pretest 1 3.42 5.65 0.16 0.09 0.02 0.24

Pretest 2 3.66 6.35 0.20 0.09 0.02 0.18

Term Day 21 3.52 5.10 0.26 0.16 0.00 0.17

Term Day 28 5.19 4.76 0.43 0.26 0.01 0.07

3146 Pretest 1 5.72 6.79 0.24 0.20 0.02 0.20

Pretest 2 8.70 5.28 0.21 0.09 0.01 0.24

Term Day 21 5.31 5.40 0.29 0.23 0.01 0.19 CLSE

Term Day 28 3.42 5.36 0.29 0.28 0.02 0.04

227
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 3

3147 Pretest 1 12.7 39.7 8.57 289.4 617 46.3 14.9 32.1 19.2 10.15

Pretest 2 12.6 38.8 8.50 200.6 578 45.7 14.9 32.6 18.6 8.33

Term Day 21 13.5 40.5 8.92 157.2 557 45.4 15.1 33.3 18.6 7.64

Term Day 28 14.6 44.6 9.52 121.4 508 46.9 15.4 32.8 18.9 8.77

3148 Pretest 1 10.7 33.0 7.13 222.5 688 46.3 15.0 32.5 18.5 13.92

Pretest 2 9.9 31.8 6.25 709.7 687 50.8 15.8 31.1 24.5 12.26

Term Day 21 10.3 31.5 6.63 105.6 580 47.5 15.6 32.7 18.3 13.48

Term Day 28 11.9 36.5 7.59 112.6 556 48.1 15.6 32.5 18.1 12.91

3149 Pretest 1 11.2 34.3 8.23 354.2 614 41.7 13.6 32.6 21.4 13.37

Pretest 2 11.5 35.2 8.49 292.0 571 41.5 13.6 32.7 19.9 10.55

Term Day 21 12.1 36.6 8.88 238.3 422 41.3 13.7 33.1 18.3 11.96

Term Day 28 12.6 38.8 9.26 239.3 448 41.9 13.6 32.4 18.6 14.48

3150 Pretest 1 13.4 41.5 7.47 136.4 710 55.5 17.9 32.2 16.1 12.65

Pretest 2 14.1 42.9 7.76 183.6 546 55.3 18.2 32.8 15.8 8.70

Term Day 21 14.3 43.1 8.10 102.9 541 53.3 17.6 33.1 15.3 8.98

Term Day 28 14.9 45.1 8.27 142.8 484 54.6 18.0 33.0 15.8 8.79

3151 Pretest 1 11.8 38.3 7.33 296.5 593 52.3 16.1 30.9 19.8 15.32

Pretest 2 12.1 37.7 7.32 235.5 613 51.6 16.5 32.1 19.5 12.07

Term Day 21 13.1 39.0 7.83 121.1 477 49.8 16.8 33.7 17.2 10.18

Term Day 28 14.1 42.7 8.27 148.3 497 51.7 17.0 32.9 17.4 12.75

228
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 3

3147 Pretest 1 2.85 6.73 0.24 0.11 0.03 0.20

Pretest 2 3.24 4.64 0.22 0.07 0.01 0.15

Term Day 21 3.46 3.68 0.28 0.12 0.01 0.10

Term Day 28 3.34 4.79 0.39 0.21 0.01 0.03

3148 Pretest 1 5.21 8.19 0.34 0.11 0.02 0.05

Pretest 2 5.43 6.41 0.15 0.07 0.01 0.19

Term Day 21 5.70 6.99 0.32 0.21 0.01 0.25

Term Day 28 4.45 7.80 0.36 0.21 0.02 0.07

3149 Pretest 1 4.83 8.02 0.16 0.13 0.01 0.22

Pretest 2 4.34 5.74 0.16 0.12 0.01 0.18

Term Day 21 5.04 6.21 0.18 0.20 0.01 0.32

Term Day 28 7.89 5.82 0.35 0.30 0.01 0.10

3150 Pretest 1 3.78 8.28 0.26 0.12 0.02 0.19

Pretest 2 2.13 6.17 0.19 0.09 0.03 0.09

Term Day 21 2.33 6.12 0.30 0.11 0.01 0.10

Term Day 28 2.20 6.18 0.23 0.15 0.02 0.02

3151 Pretest 1 5.71 8.85 0.24 0.17 0.02 0.33

Pretest 2 5.00 6.71 0.15 0.06 0.01 0.15

Term Day 21 3.75 5.90 0.18 0.15 0.01 0.20

Term Day 28 4.62 7.56 0.32 0.17 0.02 0.07

229
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 3

3152 Pretest 1 11.7 36.7 7.81 202.6 537 46.9 14.9 31.8 18.9 13.52

Pretest 2 12.1 38.6 7.92 216.1 528 48.8 15.2 31.2 19.0 8.90

Term Day 21 12.0 36.0 7.76 57.1 413 46.3 15.5 33.4 17.8 12.72

Term Day 28 13.0 40.1 8.45 101.0 417 47.4 15.4 32.4 18.2 9.40

230
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 3

3152 Pretest 1 5.67 7.02 0.31 0.19 0.02 0.29

Pretest 2 3.29 5.12 0.20 0.10 0.01 0.18

Term Day 21 6.42 5.61 0.25 0.16 0.08 0.22

Term Day 28 3.54 5.14 0.38 0.27 0.02 0.06

231
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

1645 Pretest 1 13.8 43.1 8.09 263.5 599 53.3 17.1 32.1 16.4 15.26

Pretest 2 13.1 40.2 7.71 173.0 552 52.1 17.0 32.6 16.0 12.50

Term Day 21 14.3 41.9 8.23 133.1 489 50.9 17.4 34.2 15.8 11.19

Term Day 28 14.1 39.5 7.34 179.0 483 53.8 19.2 35.6 18.6 12.52

1646 Pretest 1 13.1 40.4 8.52 376.5 693 47.4 15.3 32.3 18.3 13.93

Pretest 2 13.0 41.9 8.44 344.5 763 49.7 15.3 30.9 18.2 11.44

Term Day 21 13.2 40.0 8.53 214.6 622 46.9 15.4 32.8 17.0 10.79

Term Day 28 13.8 41.9 9.04 134.2 493 46.4 15.2 32.8 16.5 10.50

1647 Pretest 1 13.5 41.8 7.10 208.7 750 58.8 19.0 32.2 16.0 14.13

Pretest 2 13.2 40.8 6.96 166.7 724 58.5 18.9 32.3 15.5 12.18

Term Day 21 14.0 42.0 7.30 136.2 646 57.6 19.2 33.3 15.0 10.30

Term Day 28 13.9 42.6 7.19 188.5 630 59.3 19.4 32.7 16.3 11.90

1648 Pretest 1 14.0 43.6 8.20 181.0 484 53.2 17.0 32.0 16.2 14.99

Pretest 2 13.3 42.3 7.88 209.7 551 53.7 16.9 31.4 16.3 9.65

Term Day 21 14.3 43.5 8.09 157.3 428 53.8 17.7 32.9 15.4 10.09

Term Day 28 15.0 46.6 8.72 137.6 539 53.4 17.2 32.1 15.2 11.03

1649 Pretest 1 11.7 37.1 8.09 253.9 643 45.8 14.4 31.5 18.6 12.19

Pretest 2 11.5 35.4 7.74 198.3 705 45.7 14.8 32.5 18.6 10.07

Term Day 21 13.3 39.7 8.59 199.3 530 46.3 15.5 33.4 19.2 8.83

Term Day 28 13.1 39.7 8.44 186.1 561 47.1 15.5 33.0 19.5 8.21

232
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

1645 Pretest 1 1.71 12.54 0.35 0.24 0.03 0.39

Pretest 2 1.55 10.10 0.33 0.22 0.01 0.30

Term Day 21 1.73 8.65 0.30 0.26 0.01 0.23

Term Day 28 2.00 9.73 0.41 0.28 0.02 0.07

1646 Pretest 1 4.20 9.16 0.16 0.13 0.02 0.27

Pretest 2 2.62 8.38 0.16 0.08 0.01 0.18

Term Day 21 2.58 7.56 0.18 0.17 0.02 0.29

Term Day 28 1.83 8.10 0.27 0.19 0.06 0.05

1647 Pretest 1 3.13 10.29 0.16 0.23 0.02 0.30

Pretest 2 2.64 8.96 0.24 0.14 0.03 0.17

Term Day 21 1.84 7.84 0.17 0.27 0.01 0.17

Term Day 28 2.77 8.49 0.27 0.30 0.02 0.05

1648 Pretest 1 3.31 10.56 0.37 0.23 0.16 0.36

Pretest 2 1.75 7.30 0.19 0.07 0.01 0.32

Term Day 21 1.82 7.42 0.33 0.22 0.02 0.29

Term Day 28 2.84 7.45 0.43 0.22 0.05 0.05

1649 Pretest 1 4.03 7.58 0.18 0.08 0.01 0.31

Pretest 2 3.78 5.82 0.16 0.06 0.00 0.25

Term Day 21 1.96 6.32 0.14 0.10 0.11 0.21

Term Day 28 1.90 5.73 0.26 0.22 0.01 0.09

233
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 1

1650 Pretest 1 13.3 40.1 6.78 179.7 521 59.1 19.7 33.3 15.4 16.18

Pretest 2 12.9 38.5 6.61 141.0 633 58.2 19.6 33.7 15.8 12.45

Term Day 21 13.5 39.2 6.82 76.9 506 57.4 19.8 34.5 14.5 12.46

Term Day 28 13.1 38.3 6.67 55.3 476 57.4 19.6 34.1 14.8 12.28

1651 Pretest 1 12.4 38.7 8.80 312.7 708 44.0 14.1 32.0 18.0 12.01

Pretest 2 11.9 37.4 8.45 304.4 629 44.3 14.0 31.7 18.2 12.15

Term Day 21 12.7 39.2 8.82 334.9 569 44.4 14.3 32.3 18.4 9.51

Term Day 28 12.5 39.4 8.79 323.0 565 44.8 14.2 31.8 18.4 9.76

1652 Pretest 1 12.9 39.2 8.23 155.1 747 47.6 15.6 32.8 18.0 10.86

Pretest 2 12.6 38.3 8.27 98.8 720 46.3 15.2 32.8 17.5 9.37

Term Day 21 12.3 36.5 8.05 87.2 694 45.4 15.3 33.7 17.6 9.48

Term Day 28 11.6 35.1 7.28 223.6 737 48.1 15.9 33.1 21.6 9.88

Group 2

2645 Pretest 1 11.7 36.0 8.56 311.2 737 42.1 13.7 32.6 17.3 9.91

Pretest 2 10.3 31.7 7.56 270.1 779 41.9 13.7 32.6 18.9 7.87

Term Day 21 11.7 35.2 8.63 178.7 701 40.8 13.6 33.4 16.8 9.94

Term Day 28 11.7 35.2 8.40 151.5 730 41.9 13.9 33.1 18.4 10.01

234
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 1

1650 Pretest 1 4.24 10.78 0.34 0.39 0.07 0.35

Pretest 2 3.33 8.34 0.25 0.18 0.01 0.33

Term Day 21 2.74 8.49 0.34 0.47 0.02 0.39

Term Day 28 2.69 8.24 0.62 0.57 0.02 0.14

1651 Pretest 1 2.69 8.72 0.17 0.14 0.02 0.27

Pretest 2 3.16 8.37 0.26 0.12 0.06 0.19

Term Day 21 2.04 7.06 0.24 0.14 0.01 0.03

Term Day 28 1.96 7.29 0.22 0.22 0.04 0.05

1652 Pretest 1 1.67 8.66 0.13 0.18 0.02 0.22

Pretest 2 1.58 7.29 0.15 0.11 0.02 0.22

Term Day 21 1.39 7.50 0.17 0.20 0.01 0.21

Term Day 28 2.44 6.83 0.23 0.30 0.01 0.07

Group 2

2645 Pretest 1 2.06 7.22 0.26 0.19 0.01 0.16

Pretest 2 2.00 5.39 0.25 0.07 0.00 0.15

Term Day 21 2.62 6.64 0.26 0.27 0.01 0.14

Term Day 28 2.08 7.30 0.30 0.26 0.03 0.05

235
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 2

2646 Pretest 1 12.3 39.1 7.79 310.2 628 50.2 15.7 31.4 19.3 9.17

Pretest 2 11.9 36.9 7.51 211.3 625 49.2 15.8 32.1 18.4 6.89

Term Day 21 12.4 36.7 7.75 133.4 524 47.3 15.9 33.7 17.6 8.52

Term Day 28 13.1 40.2 8.17 143.8 534 49.2 16.1 32.6 17.8 9.50

2647 Pretest 1 12.6 38.6 8.62 278.8 304 44.8 14.6 32.6 18.5 12.54

Pretest 2 11.9 35.9 8.07 271.8 496 44.4 14.7 33.1 19.3 12.64

Term Day 21 12.3 35.5 7.98 225.4 357 44.5 15.4 34.6 19.8 10.73

Term Day 28 12.3 37.2 8.30 104.4 461 44.8 14.9 33.2 17.4 10.72

2648 Pretest 1 12.1 36.7 8.02 259.2 724 45.8 15.1 32.9 19.5 8.82

Pretest 2 12.2 37.3 7.80 356.7 694 47.8 15.6 32.7 20.7 6.92

Term Day 21 13.0 38.8 8.31 87.3 551 46.7 15.6 33.5 17.6 8.93

Term Day 28 12.6 37.6 8.08 95.3 526 46.5 15.6 33.6 17.7 6.73

2649 Pretest 1 13.4 41.1 6.83 247.4 616 60.2 19.6 32.5 16.3 9.13

Pretest 2 14.0 42.6 7.25 164.9 747 58.8 19.3 32.8 15.4 8.17

Term Day 21 14.4 42.2 7.42 146.4 275 56.9 19.4 34.2 16.1 7.10

Term Day 28 14.3 42.0 7.30 116.1 609 57.4 19.5 34.0 15.9 8.25

2650 Pretest 1 13.2 41.7 7.70 243.6 636 54.1 17.2 31.7 16.3 11.45

Pretest 2 13.1 40.4 7.66 208.5 562 52.8 17.0 32.3 16.5 10.08

Term Day 21 14.2 42.6 8.00 129.4 516 53.3 17.8 33.3 15.8 9.44

Term Day 28 13.7 40.9 7.81 87.8 548 52.3 17.6 33.6 16.4 9.26

236
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 2

2646 Pretest 1 2.26 6.19 0.15 0.20 0.01 0.37

Pretest 2 1.32 5.16 0.13 0.08 0.01 0.18

Term Day 21 2.02 6.00 0.12 0.17 0.01 0.19

Term Day 28 1.68 7.28 0.18 0.24 0.03 0.09

2647 Pretest 1 3.57 8.20 0.26 0.22 0.02 0.26 NCLP

Pretest 2 2.50 9.38 0.25 0.20 0.01 0.30

Term Day 21 2.55 7.56 0.15 0.23 0.01 0.21

Term Day 28 2.14 7.93 0.32 0.32 0.01 0.01

2648 Pretest 1 2.00 6.25 0.25 0.11 0.01 0.20

Pretest 2 1.21 5.37 0.11 0.06 0.01 0.15

Term Day 21 2.12 6.21 0.34 0.21 0.01 0.03

Term Day 28 2.05 4.25 0.20 0.19 0.01 0.03

2649 Pretest 1 2.15 6.44 0.20 0.17 0.02 0.15

Pretest 2 1.65 6.11 0.15 0.08 0.06 0.14

Term Day 21 2.61 4.10 0.15 0.13 0.01 0.10 CLSE

Term Day 28 2.36 5.42 0.22 0.20 0.02 0.03

2650 Pretest 1 3.83 6.78 0.40 0.19 0.01 0.24

Pretest 2 3.80 5.69 0.27 0.08 0.01 0.23

Term Day 21 3.24 5.58 0.39 0.16 0.01 0.05

Term Day 28 3.48 5.21 0.29 0.20 0.01 0.06

237
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 2

2651 Pretest 1 13.4 41.1 8.06 402.6 660 51.0 16.6 32.6 18.1 13.78

Pretest 2 12.8 38.6 7.74 282.1 506 49.9 16.5 33.0 17.3 10.06

Term Day 21 13.9 42.7 8.34 311.3 260 51.1 16.7 32.7 17.1 12.52

Term Day 28 13.0 39.0 7.76 200.8 607 50.3 16.7 33.3 17.2 11.15

2652 Pretest 1 11.9 35.8 7.15 380.9 698 50.0 16.6 33.3 18.2 14.42

Pretest 2 12.1 37.1 7.40 283.2 722 50.2 16.3 32.5 17.3 9.26

Term Day 21 12.5 36.9 7.45 202.2 605 49.5 16.8 34.0 17.0 10.77

Term Day 28 12.4 38.4 7.57 189.5 590 50.7 16.4 32.3 17.1 11.46

Group 3

3645 Pretest 1 11.6 36.0 7.57 253.2 665 47.5 15.3 32.3 18.5 12.78

Pretest 2 11.6 36.9 7.73 204.4 674 47.7 15.0 31.5 18.1 9.49

Term Day 21 11.6 35.9 7.41 204.5 480 48.4 15.6 32.3 19.1 11.61

Term Day 28 11.9 36.7 7.57 162.5 478 48.5 15.7 32.4 19.6 9.04

3646 Pretest 1 14.1 43.2 8.44 317.0 569 51.2 16.7 32.6 17.1 12.58

Pretest 2 13.5 41.1 8.20 278.9 597 50.2 16.5 32.9 17.7 8.52

Term Day 21 13.9 41.3 8.17 167.4 512 50.5 17.0 33.6 17.7 10.52

Term Day 28 14.7 43.2 8.54 198.8 339 50.5 17.2 34.1 18.4 9.98

238
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 2

2651 Pretest 1 1.81 11.00 0.29 0.32 0.02 0.33

Pretest 2 1.24 8.23 0.41 0.15 0.01 0.03

Term Day 21 1.84 9.91 0.25 0.32 0.02 0.18 CLSE

Term Day 28 2.06 8.41 0.27 0.36 0.02 0.03

2652 Pretest 1 3.74 9.65 0.42 0.15 0.02 0.45

Pretest 2 1.46 7.30 0.23 0.06 0.02 0.18

Term Day 21 2.67 7.52 0.17 0.14 0.01 0.27

Term Day 28 3.01 7.89 0.30 0.19 0.02 0.06

Group 3

3645 Pretest 1 2.63 9.47 0.26 0.13 0.01 0.29

Pretest 2 1.80 7.26 0.20 0.06 0.01 0.16

Term Day 21 4.30 6.78 0.13 0.20 0.01 0.19

Term Day 28 2.69 5.71 0.23 0.34 0.01 0.06

3646 Pretest 1 2.86 9.03 0.26 0.14 0.09 0.21

Pretest 2 2.02 6.22 0.10 0.04 0.02 0.12

Term Day 21 2.92 7.05 0.33 0.13 0.05 0.04

Term Day 28 3.14 6.25 0.23 0.31 0.02 0.03 CLSE

239
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 3

3647 Pretest 1 12.5 37.1 7.55 282.6 315 49.2 16.5 33.6 18.7 10.00

Pretest 2 12.8 40.4 7.87 441.4 623 51.4 16.2 31.6 19.6 10.11

Term Day 21 13.0 41.5 8.08 215.4 352 51.4 16.1 31.3 18.6 10.94

Term Day 28 12.6 38.7 7.59 206.1 382 51.0 16.7 32.7 19.2 9.36

3648 Pretest 1 13.6 39.9 7.64 239.6 561 52.3 17.8 34.0 18.9 12.73

Pretest 2 13.6 40.8 7.62 213.3 514 53.6 17.8 33.3 18.4 9.18

Term Day 21 15.4 45.8 8.00 215.7 458 57.2 19.2 33.6 17.1 9.62

Term Day 28 15.7 46.9 8.11 203.4 436 57.8 19.3 33.4 16.3 10.25

3649 Pretest 1 13.1 39.5 7.43 191.1 734 53.1 17.7 33.3 19.5 18.92

Pretest 2 13.7 42.5 7.70 283.0 708 55.2 17.8 32.2 20.4 11.32

Term Day 21 14.5 43.9 7.68 201.3 523 57.2 18.9 33.0 19.2 12.19

Term Day 28 13.8 40.9 7.13 151.9 665 57.4 19.4 33.8 19.2 10.03

3650 Pretest 1 12.5 39.9 8.88 257.8 644 44.9 14.1 31.3 17.6 18.73

Pretest 2 12.7 40.6 8.99 336.9 626 45.1 14.1 31.3 17.6 13.53

Term Day 21 12.8 41.9 9.21 294.1 631 45.5 13.9 30.5 18.4 15.02

Term Day 28 12.6 41.5 9.03 256.9 599 45.9 14.0 30.5 18.9 16.08

3651 Pretest 1 11.0 34.2 7.56 158.7 623 45.2 14.6 32.2 17.5 13.97

Pretest 2 10.5 32.7 7.04 194.3 329 46.4 14.9 32.1 18.7 12.53

Term Day 21 11.8 37.0 7.83 206.1 585 47.2 15.1 31.9 20.2 11.06

Term Day 28 12.2 38.2 7.96 148.4 508 48.0 15.3 31.9 20.4 11.03

240
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 3

3647 Pretest 1 2.73 6.57 0.26 0.25 0.01 0.18 CLSE

Pretest 2 1.87 7.75 0.21 0.08 0.01 0.17

Term Day 21 1.59 8.90 0.12 0.13 0.02 0.18

Term Day 28 2.45 6.44 0.16 0.22 0.01 0.08

3648 Pretest 1 3.27 8.63 0.32 0.24 0.03 0.24

Pretest 2 2.18 6.49 0.14 0.09 0.03 0.24

Term Day 21 2.21 6.80 0.21 0.20 0.03 0.17

Term Day 28 2.34 7.29 0.27 0.25 0.06 0.04

3649 Pretest 1 11.38 6.80 0.29 0.07 0.02 0.36

Pretest 2 3.45 7.18 0.24 0.11 0.02 0.32

Term Day 21 4.66 6.87 0.39 0.22 0.02 0.04

Term Day 28 3.43 6.04 0.26 0.27 0.01 0.02

3650 Pretest 1 3.67 14.07 0.56 0.31 0.04 0.08

Pretest 2 1.90 10.99 0.22 0.17 0.03 0.23

Term Day 21 3.22 10.70 0.28 0.56 0.03 0.23

Term Day 28 3.53 11.60 0.26 0.56 0.04 0.08

3651 Pretest 1 3.78 9.19 0.58 0.34 0.01 0.06

Pretest 2 2.07 9.67 0.47 0.13 0.14 0.05 CLSL

Term Day 21 2.08 8.25 0.21 0.28 0.02 0.22

Term Day 28 1.60 8.72 0.31 0.28 0.06 0.06

241
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion HGB HCT RBC RETIC PLT MCV MCH MCHC RDW WBC

Number g/dL % x10^6/uL x10^9/L x10^3/uL fL pg g/dL % x10^3/uL

Group 3

3652 Pretest 1 12.7 38.6 8.61 301.3 565 44.9 14.7 32.8 16.6 14.50

Pretest 2 12.9 40.5 8.93 338.2 525 45.3 14.5 31.9 17.1 9.37

Term Day 21 12.8 37.7 8.57 169.8 578 44.0 15.0 34.0 17.5 11.90

Term Day 28 12.7 37.5 8.45 129.0 620 44.4 15.0 33.8 17.5 12.56

242
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 7 (cont) Hematology - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion ANEU ALYM AMONO AEOS ABASO ALUC SC

Number x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL x10^3/uL

Group 3

3652 Pretest 1 5.16 8.44 0.46 0.20 0.02 0.22

Pretest 2 2.11 6.76 0.21 0.09 0.05 0.15

Term Day 21 4.14 6.98 0.20 0.30 0.05 0.23

Term Day 28 5.47 6.38 0.25 0.32 0.07 0.06

243
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 1

1145 Pretest 1 27 49 541

Pretest 2 36 45 920

Term Day 21 28 45 278

Term Day 28 25 43 228

1146 Pretest 1 21 54 217

Pretest 2 30 49 489

Term Day 21 36 56 330 SHI

Term Day 28 21 52 239

1147 Pretest 1 26 48 201

Pretest 2 38 41 271

Term Day 21 31 41 458

Term Day 28 25 39 321

1148 Pretest 1 28 49 574

Pretest 2 51 43 1366

Term Day 21 24 32 251

Term Day 28 47 34 478 SHI

1149 Pretest 1 18 39 128

Pretest 2 34 40 319

Term Day 21 23 38 349

Term Day 28 24 40 353

244
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 1

1145 Pretest 1 578 36.4 210 15.2 88 27.48 0.00

Pretest 2 901 61.4 553 8.6 77 26.74 0.00

Term Day 21 259 33.5 87 18.7 48 40.08 0.00

Term Day 28 221 10.3 23 11.4 25 6.32 0.00

1146 Pretest 1 218 65.0 142 13.9 30 31.54 0.00

Pretest 2 503 53.6 270 23.2 117 13.40 0.00

Term Day 21 304 44.6 136 9.6 29 52.52 0.00

Term Day 28 220 38.8 85 22.7 50 9.98 0.00

1147 Pretest 1 192 70.3 135 11.3 22 14.32 0.00

Pretest 2 260 18.3 48 11.1 29 14.24 0.00

Term Day 21 425 80.0 340 7.0 30 55.06 0.00

Term Day 28 309 51.4 159 10.5 32 11.94 0.00

1148 Pretest 1 585 72.5 424 9.4 55 24.86 0.00

Pretest 2 1333 73.4 978 19.9 265 102.60 0.00

Term Day 21 245 54.6 134 17.4 43 15.56 0.01

Term Day 28 447 24.0 107 18.2 81 89.60 0.00

1149 Pretest 1 120 49.7 60 21.4 26 16.86 0.00

Pretest 2 299 74.4 222 15.0 45 15.78 0.00

Term Day 21 328 41.0 134 13.2 43 9.44 0.00

Term Day 28 342 19.9 68 19.6 67 18.16 0.00

245
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 1

1150 Pretest 1 28 38 273

Pretest 2 45 43 853

Term Day 21 35 43 661

Term Day 28 26 40 250

1151 Pretest 1 26 52 405

Pretest 2 36 50 457

Term Day 21 29 47 846

Term Day 28 22 47 277

1152 Pretest 1 33 45 683

Pretest 2 36 50 364

Term Day 21 26 45 216

Term Day 28 32 48 263

Group 2

2145 Pretest 1 24 37 391

Pretest 2 24 36 403

Term Day 21 25 36 274

Term Day 28 25 37 490

246
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 1

1150 Pretest 1 283 69.4 196 9.8 28 22.62 0.00

Pretest 2 833 77.3 644 8.9 74 57.20 0.00

Term Day 21 638 56.1 358 9.8 63 91.04 0.00

Term Day 28 239 24.9 60 14.9 36 29.18 0.00

1151 Pretest 1 418 75.6 316 7.8 33 14.78 0.00

Pretest 2 447 37.3 167 19.3 86 16.84 0.00

Term Day 21 805 86.7 698 2.8 23 9.84 0.00

Term Day 28 265 29.6 78 21.1 56 13.84 0.00

1152 Pretest 1 701 71.5 501 6.2 43 56.96 0.00

Pretest 2 362 33.5 121 21.8 79 25.42 0.00

Term Day 21 208 31.2 65 15.9 33 14.08 0.00

Term Day 28 248 22.9 57 15.1 37 20.66 0.00

Group 2

2145 Pretest 1 385 50.0 193 12.3 47 27.08 0.00

Pretest 2 385 47.3 182 15.5 60 14.82 0.00

Term Day 21 253 31.7 80 14.0 35 14.80 0.00

Term Day 28 458 24.1 110 19.0 87 97.36 0.00

247
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 2

2146 Pretest 1 29 45 350

Pretest 2 33 46 310

Term Day 21 33 40 351

Term Day 28 26 41 239

2147 Pretest 1 73 44 2486

Pretest 2 29 41 1035

Term Day 21 NS NS NS

Term Day 28 NS NS NS

Unscheduled 32 38 1185

2149 Pretest 1 31 45 324

Pretest 2 22 41 209

Term Day 21 24 41 365

Term Day 28 22 45 220

2150 Pretest 1 37 38 969

Pretest 2 30 33 476

Term Day 21 30 31 279

Term Day 28 30 34 469

248
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 2

2146 Pretest 1 343 25.9 89 8.0 27 57.28 0.00

Pretest 2 288 44.3 128 10.6 31 6.52 0.00

Term Day 21 334 60.7 203 10.9 36 31.50 0.00

Term Day 28 226 25.0 57 13.1 30 19.64 0.00

2147 Pretest 1 2387 72.7 1735 9.3 222 422.60 0.00

Pretest 2 974 71.2 693 9.5 93 12.22 0.01

Unscheduled 1100 54.4 598 20.5 226 136.48 0.00

2149 Pretest 1 310 56.3 175 14.1 44 22.00 0.00

Pretest 2 202 56.0 113 13.9 28 12.88 0.00

Term Day 21 340 91.7 312 5.9 20 53.16 0.00

Term Day 28 211 34.5 73 12.4 26 30.96 0.00

2150 Pretest 1 983 68.3 671 12.1 119 59.60 0.00

Pretest 2 444 52.9 235 8.2 36 43.44 0.00

Term Day 21 265 82.3 218 9.3 25 19.58 0.00

Term Day 28 446 44.7 199 9.8 44 21.06 0.00

249
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 2

2151 Pretest 1 23 40 291

Pretest 2 59 41 1713

Term Day 21 23 38 279

Term Day 28 18 38 239

2152 Pretest 1 22 36 505

Pretest 2 23 34 343

Term Day 21 28 35 355

Term Day 28 26 34 270

2153 Pretest 1 17 36 133

Pretest 2 21 33 225

Term Day 21 20 33 209

Term Day 28 32 34 1797

Group 3

3145 Pretest 1 29 46 337

Pretest 2 52 46 1830

Term Day 21 36 41 322

Term Day 28 27 43 216

250
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 2

2151 Pretest 1 279 42.0 117 20.7 58 20.18 0.00

Pretest 2 1558 68.7 1070 9.4 146 270.80 0.00

Term Day 21 262 74.0 194 16.6 43 13.22 0.00

Term Day 28 229 24.1 55 13.4 31 9.62 0.00

2152 Pretest 1 500 61.1 306 13.6 68 18.80 0.00

Pretest 2 324 54.9 178 12.1 39 7.48 0.00

Term Day 21 326 77.1 251 12.7 41 24.74 0.00

Term Day 28 255 42.1 107 14.3 36 13.96 0.00

2153 Pretest 1 133 27.7 37 19.8 26 4.68 0.00

Pretest 2 216 50.0 108 12.0 26 7.56 0.00

Term Day 21 201 18.0 36 13.4 27 20.28 0.00

Term Day 28 1604 63.2 1014 9.1 146 110.88 0.00

Group 3

3145 Pretest 1 320 32.9 105 27.1 87 10.76 0.00

Pretest 2 1679 55.2 927 9.6 161 73.40 0.00

Term Day 21 302 57.7 174 14.7 44 24.44 0.00

Term Day 28 202 25.7 52 21.6 44 11.08 0.00

251
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 3

3146 Pretest 1 27 48 189

Pretest 2 37 49 279

Term Day 21 28 42 190

Term Day 28 27 43 238

3147 Pretest 1 48 36 1770

Pretest 2 37 38 681

Term Day 21 27 38 265

Term Day 28 35 36 1163

3148 Pretest 1 26 36 414

Pretest 2 27 38 265

Term Day 21 25 36 422

Term Day 28 19 34 146

3149 Pretest 1 32 55 471

Pretest 2 40 54 574

Term Day 21 62 48 3200

Term Day 28 34 49 634

3150 Pretest 1 29 51 546

Pretest 2 28 49 274

Term Day 21 21 49 213

Term Day 28 21 50 238

252
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 3

3146 Pretest 1 192 31.2 60 14.4 28 13.68 0.00

Pretest 2 263 35.3 93 16.9 44 6.94 0.00

Term Day 21 173 41.4 72 11.2 19 10.72 0.00

Term Day 28 221 33.0 73 10.0 22 21.86 0.00

3147 Pretest 1 1702 70.0 1191 8.9 151 242.40 0.00

Pretest 2 662 51.1 338 17.5 116 47.60 0.00

Term Day 21 246 74.8 184 14.9 37 16.30 0.00

Term Day 28 1093 60.3 659 6.8 74 118.20 0.00

3148 Pretest 1 415 49.5 205 12.1 50 47.84 0.01

Pretest 2 252 52.0 131 10.0 25 5.54 0.01

Term Day 21 406 75.1 305 9.8 40 42.82 0.03

Term Day 28 133 28.0 37 11.0 15 9.30 0.03

3149 Pretest 1 477 23.0 110 6.8 32 55.12 0.00

Pretest 2 529 52.7 279 9.4 50 46.00 0.00

Term Day 21 2751 94.1 2589 1.4 39 272.80 0.00

Term Day 28 594 29.0 172 9.2 55 117.92 0.00

3150 Pretest 1 552 38.2 211 12.9 71 37.12 0.00

Pretest 2 254 49.8 126 12.6 32 24.68 0.00

Term Day 21 198 73.4 145 12.5 25 15.06 0.00

Term Day 28 219 33.8 74 15.3 34 11.40 0.00

253
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 3

3151 Pretest 1 46 47 1941

Pretest 2 33 47 347

Term Day 21 33 51 631

Term Day 28 45 57 2367

3152 Pretest 1 24 44 257

Pretest 2 33 50 371

Term Day 21 26 43 231

Term Day 28 22 41 277

254
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Males

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 3

3151 Pretest 1 1807 63.4 1146 8.0 145 97.80 0.00

Pretest 2 328 53.7 176 10.3 34 17.78 0.00

Term Day 21 609 88.6 540 6.4 39 63.20 0.00

Term Day 28 2060 69.2 1426 9.6 198 101.60 0.00

3152 Pretest 1 264 32.7 86 14.0 37 9.60 0.00

Pretest 2 341 57.8 197 12.8 44 10.82 0.00

Term Day 21 214 85.5 183 9.1 19 15.42 0.00

Term Day 28 259 31.3 81 10.5 27 10.10 0.00

255
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 1

1645 Pretest 1 25 41 324 SHI

Pretest 2 20 39 420

Term Day 21 22 39 678

Term Day 28 24 42 526

1646 Pretest 1 25 51 569

Pretest 2 19 47 344

Term Day 21 19 46 974

Term Day 28 18 46 477

1647 Pretest 1 31 44 473

Pretest 2 31 42 516

Term Day 21 33 37 1263

Term Day 28 27 40 1004

1648 Pretest 1 35 43 353 SHI

Pretest 2 22 41 453

Term Day 21 34 42 756

Term Day 28 33 37 856 SHI

1649 Pretest 1 23 31 925

Pretest 2 29 32 7431

Term Day 21 25 36 1862

Term Day 28 26 37 1290

256
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 1

1645 Pretest 1 420 39.2 165 12.4 52 10.46 0.00

Pretest 2 405 84.7 343 2.9 12 9.66 0.00

Term Day 21 648 90.4 586 6.1 40 22.06 0.00

Term Day 28 489 49.8 244 10.5 51 14.18 0.00

1646 Pretest 1 584 42.3 247 17.5 102 27.70 0.00

Pretest 2 332 85.1 283 5.4 18 6.50 0.00

Term Day 21 927 95.5 885 3.8 35 7.74 0.00

Term Day 28 448 31.5 141 11.3 51 15.46 0.00

1647 Pretest 1 490 46.5 228 17.7 87 9.08 1.68

Pretest 2 491 78.2 384 6.2 30 12.88 2.52

Term Day 21 1181 91.2 1077 5.3 63 19.44 3.92

Term Day 28 935 53.2 497 15.2 142 14.34 4.13

1648 Pretest 1 420 68.3 287 11.8 50 14.62 0.00

Pretest 2 421 72.4 305 9.2 39 49.36 0.00

Term Day 21 702 94.0 660 4.4 31 19.86 0.00

Term Day 28 797 54.6 435 9.9 79 100.72 0.03

1649 Pretest 1 952 91.6 872 3.2 30 16.48 0.00

Pretest 2 5469 95.5 5223 2.1 115 18.80 0.00

Term Day 21 1687 88.3 1490 6.7 113 23.32 0.00

Term Day 28 1193 46.0 549 22.0 262 87.04 0.00

257
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 1

1650 Pretest 1 23 43 708

Pretest 2 33 45 955

Term Day 21 17 43 594

Term Day 28 20 42 520

1651 Pretest 1 30 39 596

Pretest 2 26 37 856

Term Day 21 39 36 516

Term Day 28 21 35 347

1652 Pretest 1 21 44 403

Pretest 2 26 44 483

Term Day 21 35 42 1794

Term Day 28 28 45 1278

Group 2

2645 Pretest 1 36 38 831

Pretest 2 27 34 227

Term Day 21 22 35 730

Term Day 28 19 37 366

258
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 1

1650 Pretest 1 712 77.0 548 9.4 67 20.82 0.00

Pretest 2 898 65.6 589 14.6 131 123.80 0.00

Term Day 21 551 75.8 418 13.6 75 18.20 0.00

Term Day 28 468 47.7 223 12.5 59 17.16 0.00

1651 Pretest 1 597 70.3 420 14.5 87 24.30 0.00

Pretest 2 819 59.2 485 16.9 138 102.56 0.00

Term Day 21 1412 95.5 1348 3.6 51 16.32 0.00

Term Day 28 317 34.4 109 24.6 78 9.26 0.00

1652 Pretest 1 407 73.9 301 13.8 56 48.00 0.00

Pretest 2 467 34.7 162 14.1 66 28.22 0.00

Term Day 21 1601 97.1 1555 2.6 42 121.92 0.01

Term Day 28 1163 65.5 762 14.0 163 73.48 0.01

Group 2

2645 Pretest 1 823 92.6 762 4.4 36 65.40 0.00

Pretest 2 209 44.2 92 10.2 21 8.60 0.00

Term Day 21 708 93.0 658 4.9 35 15.76 0.00

Term Day 28 344 45.1 155 16.3 56 12.22 0.00

259
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 2

2646 Pretest 1 33 37 803 SHI

Pretest 2 31 28 1939

Term Day 21 27 30 1961

Term Day 28 20 31 272

2647 Pretest 1 391 187 >7800

Pretest 2 209 165 >7800

Term Day 21 74 216 >7800

Term Day 28 160 219 >7800

2648 Pretest 1 25 47 298

Pretest 2 27 47 374

Term Day 21 19 44 455

Term Day 28 25 42 461

2649 Pretest 1 34 36 674

Pretest 2 28 35 480

Term Day 21 24 38 1540

Term Day 28 22 35 286

2650 Pretest 1 45 39 1554 SHI

Pretest 2 21 36 391

Term Day 21 23 42 5774

Term Day 28 18 38 722

260
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 2

2646 Pretest 1 795 71.8 571 14.4 114 51.36 0.01

Pretest 2 1760 57.7 1016 14.7 259 25.48 0.02

Term Day 21 1824 92.8 1693 5.2 95 9.18 0.03

Term Day 28 255 20.5 52 11.0 28 22.94 0.03

2647 Pretest 1 48370 96.5 46677 1.9 919 115.80 0.00

Pretest 2 17750 94.4 16756 4.1 728 1003.20 0.00

Term Day 21 18520 93.4 17298 4.2 778 117.36 0.00

Term Day 28 38330 96.1 36835 2.4 920 168.40 0.00

2648 Pretest 1 301 70.3 212 16.9 51 7.88 0.00

Pretest 2 352 33.6 118 10.4 37 30.00 0.00

Term Day 21 446 65.0 290 19.9 89 12.48 0.00

Term Day 28 434 20.7 90 8.2 36 59.98 0.00

2649 Pretest 1 31 86.1 27 8.4 3 44.96 0.00

Pretest 2 458 77.2 354 7.4 34 22.10 0.00

Term Day 21 498 79.5 396 10.8 54 17.14 0.00

Term Day 28 268 22.3 60 8.0 21 18.16 0.00

2650 Pretest 1 1490 71.7 1068 7.6 113 299.40 0.00

Pretest 2 379 71.0 269 11.4 43 18.42 0.00

Term Day 21 6920 94.5 6539 4.5 311 68.70 0.00

Term Day 28 662 27.0 179 8.6 57 7.62 0.00

261
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 2

2651 Pretest 1 25 40 451

Pretest 2 31 46 552

Term Day 21 21 35 1038

Term Day 28 36 35 885

2652 Pretest 1 18 41 231

Pretest 2 19 40 364

Term Day 21 18 36 284

Term Day 28 16 35 320

Group 3

3645 Pretest 1 21 40 365

Pretest 2 21 36 481

Term Day 21 22 37 1025

Term Day 28 30 34 982

3646 Pretest 1 37 72 887

Pretest 2 31 76 798

Term Day 21 27 59 >7800

Term Day 28 57 64 3271 SHI

262
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 2

2651 Pretest 1 441 76.8 339 17.7 78 11.58 0.00

Pretest 2 526 82.1 432 8.4 44 18.80 0.00

Term Day 21 1009 87.2 880 8.3 84 14.72 0.01

Term Day 28 824 43.9 362 9.0 74 59.02 0.00

2652 Pretest 1 224 62.5 140 16.4 37 22.50 0.30

Pretest 2 346 74.8 259 14.9 52 43.78 0.17

Term Day 21 265 83.3 221 12.5 33 8.86 0.08

Term Day 28 297 23.5 70 14.0 42 17.68 0.09

Group 3

3645 Pretest 1 356 92.6 330 4.0 14 20.28 0.00

Pretest 2 444 77.4 344 14.5 64 49.82 0.00

Term Day 21 1010 53.9 544 14.0 141 53.58 0.00

Term Day 28 909 33.1 301 8.9 81 98.88 0.00

3646 Pretest 1 892 69.2 617 6.6 59 23.12 0.00

Pretest 2 758 80.9 613 8.3 63 20.90 0.00

Term Day 21 10790 95.1 10261 1.5 162 53.44 0.00

Term Day 28 2707 57.9 1567 7.3 198 562.20 0.00

263
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 3

3647 Pretest 1 26 43 668

Pretest 2 25 45 527

Term Day 21 25 42 524

Term Day 28 22 41 564

3648 Pretest 1 29 38 1430

Pretest 2 26 41 815

Term Day 21 20 40 2246

Term Day 28 35 40 2432

3649 Pretest 1 31 39 358

Pretest 2 33 43 696

Term Day 21 24 38 >7800

Term Day 28 22 34 908

3650 Pretest 1 32 43 978 SHI

Pretest 2 22 46 310

Term Day 21 28 44 635

Term Day 28 23 40 337

3651 Pretest 1 24 37 654

Pretest 2 35 39 1272

Term Day 21 21 42 >7800

Term Day 28 24 38 586

264
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 3

3647 Pretest 1 671 72.8 488 19.6 132 33.36 0.00

Pretest 2 480 74.9 360 11.1 53 43.32 0.00

Term Day 21 512 32.0 164 8.4 43 35.88 0.00

Term Day 28 524 40.0 210 10.9 57 49.62 0.00

3648 Pretest 1 1396 87.9 1227 6.7 94 51.20 0.00

Pretest 2 760 58.2 442 14.4 109 36.78 0.00

Term Day 21 2056 62.2 1279 15.5 319 16.88 0.00

Term Day 28 2055 64.5 1325 7.8 160 508.80 0.00

3649 Pretest 1 368 62.5 230 16.4 60 7.64 0.00

Pretest 2 634 73.2 464 17.6 112 16.72 0.00

Term Day 21 18710 93.8 17550 4.4 823 28.32 0.00

Term Day 28 836 40.9 342 10.2 85 18.76 0.00

3650 Pretest 1 1007 84.7 853 7.4 75 28.86 0.00

Pretest 2 285 66.8 190 20.9 60 34.32 0.00

Term Day 21 631 26.1 165 10.0 63 14.24 0.00

Term Day 28 309 13.4 41 4.1 13 28.82 0.00

3651 Pretest 1 674 74.4 501 16.4 111 8.52 0.00

Pretest 2 1182 84.4 998 4.6 54 100.14 0.00

Term Day 21 12270 90.2 11068 4.1 503 35.94 0.00

Term Day 28 535 47.5 254 8.5 45 33.90 0.00

265
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion AST ALT CK SC

Number U/L U/L U/L

Group 3

3652 Pretest 1 66 42 1488

Pretest 2 24 41 743

Term Day 21 17 36 883

Term Day 28 17 37 630

266
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 8 (cont) Clinical chemistry - individual values

Females

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Animal Occasion CK CK(MM) CK(MM) CK(MB) CK(MB) MYOGLOBI TPI

Number U/L % U/L % U/L NG/mL NG/mL

Group 3

3652 Pretest 1 1431 83.6 1196 7.5 107 56.40 0.00

Pretest 2 672 34.4 231 16.6 112 63.36 0.00

Term Day 21 853 42.2 360 18.1 154 7.12 0.00

Term Day 28 569 26.3 150 11.0 63 37.72 0.00

267
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 9 Organ weights - individual values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group 

/Sex

Animal 

Number

Terminal 

Body weight Brain Brain Heart Heart Heart Kidneys Kidneys Kidneys

Relative to Body (%) Body (%) Brain (%) Body (%) Brain (%)

1M 1145 11300.0 56.145 0.497 68.308 0.604 122 96.366 0.853 172

1146 9300.0 53.764 0.578 48.181 0.518 90 69.044 0.742 128

1147 11500.0 59.261 0.515 58.563 0.509 99 70.882 0.616 120

1148 11800.0 62.544 0.530 65.105 0.552 104 73.724 0.625 118

1149 11300.0 56.310 0.498 65.089 0.576 116 60.046 0.531 107

1150 12400.0 58.425 0.471 77.921 0.628 133 67.398 0.544 115

1151 8700.0 53.209 0.612 46.361 0.533 87 63.053 0.725 119

1152 9700.0 58.075 0.599 50.918 0.525 88 69.113 0.713 119

2M 2145 10000.0 49.925 0.499 60.417 0.604 121 62.896 0.629 126

2146 9100.0 53.682 0.590 59.821 0.657 111 58.725 0.645 109

2149 8600.0 50.905 0.592 48.560 0.565 95 45.650 0.531 90

2150 11100.0 67.293 0.606 55.551 0.500 83 68.742 0.619 102

2151 10800.0 63.159 0.585 68.696 0.636 109 78.961 0.731 125

2152 10900.0 61.499 0.564 62.164 0.570 101 75.303 0.691 122

2153 11000.0 60.081 0.546 59.892 0.544 100 54.043 0.491 90

268
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 9 (cont) Organ weights - individual values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group 

/Sex

Animal 

Number Liver Liver Liver

Lungs and 

Bronchi

Lungs and 

Bronchi

Lungs and 

Bronchi

Relative to Body (%) Brain (%) Body (%) Brain (%)

1M 1145 300.731 2.66 536 79.082 0.700 141

1146 206.423 2.22 384 73.803 0.794 137

1147 268.473 2.33 453 100.887 0.877 170

1148 267.020 2.26 427 98.371 0.834 157

1149 218.405 1.93 388 95.507 0.845 170

1150 235.037 1.90 402 112.514 0.907 193

1151 176.556 2.03 332 73.581 0.846 138

1152 201.813 2.08 348 84.130 0.867 145

2M 2145 234.571 2.35 470 82.969 0.830 166

2146 191.784 2.11 357 83.324 0.916 155

2149 153.412 1.78 301 81.816 0.951 161

2150 279.407 2.52 415 80.554 0.726 120

2151 203.820 1.89 323 91.179 0.844 144

2152 295.471 2.71 480 99.194 0.910 161

2153 199.304 1.81 332 97.086 0.883 162

269
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 9 (cont) Organ weights - individual values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group 

/Sex

Animal 

Number

Terminal 

Body weight Brain Brain Heart Heart Heart Kidneys Kidneys Kidneys

Relative to Body (%) Body (%) Brain (%) Body (%) Brain (%)

3M 3145 8200.0 52.729 0.643 47.779 0.583 91 66.020 0.805 125

3146 8100.0 53.527 0.661 56.406 0.696 105 61.114 0.754 114

3147 10900.0 56.169 0.515 67.921 0.623 121 93.088 0.854 166

3148 11700.0 61.709 0.527 76.826 0.657 124 77.042 0.658 125

3149 8300.0 53.890 0.649 53.191 0.641 99 54.656 0.659 101

3150 9900.0 59.927 0.605 58.436 0.590 98 77.605 0.784 129

3151 12600.0 55.373 0.439 69.335 0.550 125 87.525 0.695 158

3152 9100.0 59.061 0.649 58.582 0.644 99 63.647 0.699 108

270
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 9 (cont) Organ weights - individual values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group 

/Sex

Animal 

Number Liver Liver Liver

Lungs and 

Bronchi

Lungs and 

Bronchi

Lungs and 

Bronchi

Relative to Body (%) Brain (%) Body (%) Brain (%)

3M 3145 192.425 2.35 365 83.127 1.014 158

3146 185.388 2.29 346 80.181 0.990 150

3147 257.695 2.36 459 79.237 0.727 141

3148 316.082 2.70 512 91.052 0.778 148

3149 189.563 2.28 352 67.034 0.808 124

3150 199.843 2.02 333 73.318 0.741 122

3151 321.390 2.55 580 110.988 0.881 200

3152 197.748 2.17 335 79.156 0.870 134

271
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 9 (cont) Organ weights - individual values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group 

/Sex

Animal 

Number

Terminal 

Body weight Brain Brain Heart Heart Heart Kidneys Kidneys Kidneys

Relative to Body (%) Body (%) Brain (%) Body (%) Brain (%)

1F 1645 12190.0 54.125 0.444 45.920 0.377 84.8 55.131 0.452 101.9

1646 12070.0 65.697 0.544 49.573 0.411 75.5 57.268 0.474 87.2

1647 12430.0 61.489 0.495 52.563 0.423 85.5 50.989 0.410 82.9

1648 9720.0 57.768 0.594 44.912 0.462 77.7 39.380 0.405 68.2

1649 13500.0 63.982 0.474 63.811 0.473 99.7 63.415 0.470 99.1

1650 11120.0 57.095 0.513 45.403 0.408 79.5 54.964 0.494 96.3

1651 10390.0 59.444 0.572 43.061 0.414 72.4 47.383 0.456 79.7

1652 11150.0 66.085 0.593 61.217 0.549 92.6 57.133 0.512 86.5

2F 2645 9720.0 53.390 0.549 46.269 0.476 86.7 39.017 0.401 73.1

2646 13820.0 57.386 0.415 53.981 0.391 94.1 69.130 0.500 120.5

2647 9430.0 54.758 0.581 49.153 0.521 89.8 53.766 0.570 98.2

2648 10870.0 59.925 0.551 51.374 0.473 85.7 52.227 0.480 87.2

2649 12530.0 59.725 0.477 54.158 0.432 90.7 52.535 0.419 88.0

2650 10210.0 55.881 0.547 45.187 0.443 80.9 45.933 0.450 82.2

2651 12170.0 63.728 0.524 51.262 0.421 80.4 54.831 0.451 86.0

2652 9920.0 54.535 0.550 40.896 0.412 75.0 51.401 0.518 94.3

272
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 9 (cont) Organ weights - individual values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group 

/Sex

Animal 

Number Liver Liver Liver

Lungs and 

Bronchi

Lungs and 

Bronchi

Lungs and 

Bronchi

Relative to Body (%) Brain (%) Body (%) Brain (%)

1F 1645 252.236 2.07 466 81.282 0.667 150

1646 220.168 1.82 335 77.795 0.645 118

1647 265.209 2.13 431 97.657 0.786 159

1648 211.665 2.18 366 71.000 0.730 123

1649 247.886 1.84 387 89.062 0.660 139

1650 229.358 2.06 402 78.878 0.709 138

1651 179.302 1.73 302 83.324 0.802 140

1652 183.885 1.65 278 85.568 0.767 129

2F 2645 195.165 2.01 366 74.840 0.770 140

2646 301.660 2.18 526 91.586 0.663 160

2647 239.892 2.54 438 68.762 0.729 126

2648 213.415 1.96 356 94.470 0.869 158

2649 318.409 2.54 533 93.523 0.746 157

2650 196.021 1.92 351 74.844 0.733 134

2651 294.424 2.42 462 83.438 0.686 131

2652 261.228 2.63 479 NW - -

NW - Not weighed

273
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 9 (cont) Organ weights - individual values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group 

/Sex

Animal 

Number

Terminal 

Body weight Brain Brain Heart Heart Heart Kidneys Kidneys Kidneys

Relative to Body (%) Body (%) Brain (%) Body (%) Brain (%)

3F 3645 12510.0 57.308 0.458 51.633 0.413 90.1 60.227 0.481 105.1

3646 12010.0 48.665 0.405 45.524 0.379 93.5 69.302 0.577 142.4

3647 9270.0 50.597 0.546 54.672 0.590 108.1 33.672 0.363 66.5

3648 12220.0 60.602 0.496 59.313 0.485 97.9 55.958 0.458 92.3

3649 11030.0 58.433 0.530 45.391 0.412 77.7 48.202 0.437 82.5

3650 9550.0 53.533 0.561 54.307 0.569 101.4 45.051 0.472 84.2

3651 13430.0 55.615 0.414 60.710 0.452 109.2 61.406 0.457 110.4

3652 11740.0 58.114 0.495 42.824 0.365 73.7 47.493 0.405 81.7

274
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

APPENDIX 9 (cont) Organ weights - individual values (g)

Vehicle Control HFO-1234yf

Dose Group 1 2 3

Dose (ppm) 5000 10000

Group 

/Sex

Animal 

Number Liver Liver Liver

Lungs and 

Bronchi

Lungs and 

Bronchi

Lungs and 

Bronchi

Relative to Body (%) Brain (%) Body (%) Brain (%)

3F 3645 307.481 2.46 537 98.977 0.791 173

3646 285.991 2.38 588 87.674 0.730 180

3647 222.885 2.40 441 79.615 0.859 157

3648 267.733 2.19 442 88.356 0.723 146

3649 264.183 2.40 452 90.383 0.819 155

3650 201.229 2.11 376 78.417 0.821 146

3651 358.243 2.67 644 105.675 0.787 190

3652 268.751 2.29 462 65.975 0.562 114

275
Sponsor refe

Huntingdo

APPENDIX 10 Macropathology - individual findings  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Day (Week) 

/Sex Number of Death Phase Tissue Findings 

1M 1145S1 29 (5) Treatment Lungs and Bronchi Dark area(s), right cranial lobe, red, diffu

Remainder of animal No macropathology findings 

 1146S1 29 (5) Treatment All tissues No macropathology findings  1147S1 29 (5) Treatment All tissues No macropathology findings  1148S1 29 (5) Treatment All tissues No macropathology findings  1149S1 29 (5) Treatment All tissues No macropathology findings  1150S1 29 (5) Treatment All tissues No macropathology findings  1151S1 29 (5) Treatment All tissues No macropathology findings  1152S1 29 (5) Treatment All tissues No macropathology findings 

2M 2145S1 29 (5) Treatment All tissues No macropathology findings 

S1 - Scheduled Euthanasia 1 

276
Sponsor refe

Huntingdo

APPENDIX 10 (cont) Macropathology - individual findings  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Day (Week) 

/Sex Number of Death Phase Tissue Findings 

2M 2146S1 29 (5) Treatment All tissues No macropathology findings 

 2147WE 16 (3) Treatment Colon Abnormal contents, material, clear, gelat

Heart Thrombus, left ventricle, red, 10x30mm, 

Thrombus, right ventricle, red, 20x40mm

 Rectum Prolapse, noted 

Remainder of animal No macropathology findings 

 2149S1 29 (5) Treatment All tissues No macropathology findings  2150S1 29 (5) Treatment All tissues No macropathology findings  2151S1 29 (5) Treatment All tissues No macropathology findings  2152S1 29 (5) Treatment All tissues No macropathology findings 

S1 - Scheduled Euthanasia 1, WE - Euthanized for welfare reasons 

277
Sponsor refe

Huntingdo

APPENDIX 10 (cont) Macropathology - individual findings  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Day (Week) 

/Sex Number of Death Phase Tissue Findings 

2M 2153S1 29 (5) Treatment All tissues No macropathology findings 3M 3145S1 29 (5) Treatment All tissues No macropathology findings 

 3146S1 29 (5) Treatment All tissues No macropathology findings  3147S1 29 (5) Treatment All tissues No macropathology findings  3148S1 29 (5) Treatment Heart Pericaridial Fluid, clear (increased.) 

Remainder of animal No macropathology findings 

 3149S1 29 (5) Treatment All tissues No macropathology findings  3150S1 29 (5) Treatment All tissues No macropathology findings  3151S1 29 (5) Treatment Lungs and Bronchi Dark area(s), all lobes, red, </=1mm, 5+ 

Remainder of animal No macropathology findings 

S1 - Scheduled Euthanasia 1 

278
Sponsor refe

Huntingdo

APPENDIX 10 (cont) Macropathology - individual findings  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Day (Week) 

/Sex Number of Death Phase Tissue Findings 

3M 3152S1 29 (5) Treatment Heart Adhesion(s), pericardium, focal, left, not

ventricle.) 

Rectum Dark area(s), mucosa, lower, red, </=1m

Remainder of animal No macropathology findings 

S1 - Scheduled Euthanasia 1 

279
Sponsor refe

Huntingdo

APPENDIX 10 (cont) Macropathology - individual findings  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Day (Week) 

/Sex Number of Death Phase Tissue Findings 

1F 1645S1 29 (5) Treatment All tissues No macropathology findings 

 1646S1 29 (5) Treatment All tissues No macropathology findings  1647S1 29 (5) Treatment All tissues No macropathology findings  1648S1 29 (5) Treatment All tissues No macropathology findings  1649S1 29 (5) Treatment All tissues No macropathology findings  1650S1 29 (5) Treatment All tissues No macropathology findings  1651S1 29 (5) Treatment All tissues No macropathology findings  1652S1 29 (5) Treatment Kidneys Cyst(s), caudal pole, right, clear, 10-19m

Remainder of animal No macropathology findings 

2F 2645S1 29 (5) Treatment All tissues No macropathology findings 

S1 - Scheduled Euthanasia 1 

280
Sponsor refe

Huntingdo

APPENDIX 10 (cont) Macropathology - individual findings  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Day (Week) 

/Sex Number of Death Phase Tissue Findings 

2F 2646S1 29 (5) Treatment All tissues No macropathology findings 

 2647S1 29 (5) Treatment All tissues No macropathology findings  2648S1 29 (5) Treatment Stomach Dark area(s), cardiac region, mucosa, red

Remainder of animal No macropathology findings 

 2649S1 29 (5) Treatment Lungs and Bronchi Dark area(s), all lobes, 2-9mm, 5+ (many

Remainder of animal No macropathology findings 

 2650S1 29 (5) Treatment All tissues No macropathology findings  2651S1 29 (5) Treatment Lymph Node, Mandibular Dark, bilateral 

Remainder of animal No macropathology findings 

 2652S1 29 (5) Treatment All tissues No macropathology findings 

S1 - Scheduled Euthanasia 1 

281
Sponsor refe

Huntingdo

APPENDIX 10 (cont) Macropathology - individual findings  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Day (Week) 

/Sex Number of Death Phase Tissue Findings 

3F 3645S1 29 (5) Treatment All tissues No macropathology findings 

 3646S1 29 (5) Treatment Lungs and Bronchi Dark area(s), all lobes, red, 2-9mm, 5+ (

Remainder of animal No macropathology findings 

 3647S1 29 (5) Treatment All tissues No macropathology findings  3648S1 29 (5) Treatment Lymph Node, Mandibular Dark, bilateral 

Remainder of animal No macropathology findings 

 3649S1 29 (5) Treatment All tissues No macropathology findings  3650S1 29 (5) Treatment All tissues No macropathology findings  3651S1 29 (5) Treatment Lungs and Bronchi Dark area(s), all lobes, red, </=1mm, not

S1 - Scheduled Euthanasia 1 

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APPENDIX 10 (cont) Macropathology - individual findings  Vehicle Control HFO-1234yf Dose Group 1 2 3 Dose (ppm) 5000 10000 

Group Animal Day (Week) 

/Sex Number of Death Phase Tissue Findings 

3F 3651S1 29 (5) Treatment Stomach Dark area(s), cardiac region, mucosa, red

Dark area(s), pylorus, mucosa, red, 2-9m

Remainder of animal No macropathology findings 

 3652S1 29 (5) Treatment All tissues No macropathology findings 

S1 - Scheduled Euthanasia 1 

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Annex 1 Certificates of analysis 

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Annex 2 Aerosol technology report

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1

Working for a better future

Aerosol Technology Report

HLS study number: 13-6434 

Sponsor study number:

Version: Final

Date: 17 January 2014

296
SIGNATURE PAGE 

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Huntingdon Life Sciences: 13-6434 

The following scientist was responsible for this portion of the study: 

Date 

2 

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1. Pre-study Atmosphere Characterization

Inhalation exposure system characterized including determinations of:

• Achieved concentration

• Homogeneity of test substance distribution in the chamber

2. Test Substance Handling and Processing for Atmosphere

Generation 

As supplied, HFO-1234yf metered directly from Sponsor cylinder as a gas.

3. Exposure System

A schematic of the exposure system is presented in Figure 1. Components of the 

exposure system:

• Whole-body exposure chambers: 

o Control Group – 1500 L internal volume

o Treated Groups – 1000 L internal volume

o Five separate chambers for Group 1

 Two chambers for males and two for females (Days 1 to 17) 

 Three chambers for males and two for females (Days 18 to 28) 

o Four separate chambers for each of Groups 2 and 3

 Two chambers for males and two for females 

• Animal Restraint:

o No direct restraint employed, animals free to move within the containment 

volume 

o Inhalation exposure chamber fitted with perforated rubber matting over a 

stainless steel floor grating

o Group housed

 4 animals/sex/chamber

• All Group 1 Female and two Group 1 Male chamber Days 1 

to 17

• All Group 2 Female and one Group 2 Male chamber

• All Group 3 Male and Female chambers

 3 animals/sex/chamber – single Group 2 Male chamber 

 1 animal/sex/chamber – single Group 1 Male chamber Days 18 to 

28

o Animal restrained separate from others for welfare reasons in each of 

Group 1 (male) and Group 2 (male)

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o Chamber identification was as follows:

 Group 1: 101M (3 animals only starting Day 18), 102M, 101F, 102F

 Group 2: 201M (3 animals only starting Day 16), 202M, 201F, 202F

 Group 3: 301M, 302M, 301F, 302F

 An additional chamber introduced (103M) on Exposure Day 18 to 

permit separation of one animal for welfare reasons

• Atmosphere Generation System: 

o Single Sponsor cylinder for each of Groups 2 and 3

o Test substance metered as gas directly from a central Sponsor supplied 

cylinder to separate Mass Flowmeters for each male and female exposure 

chamber

o Test substance flow monitored continuously using Mass Flowmeter

displays and adjusted as required based on Infrared Gas Analyzer

indicated concentrations 

o Group 1 exposed to room air only 

o Make-up air for Groups 2 and 3 drawn passively from room environment 

by calibrated chamber exhaust

• Extract Airflow:

o Drawn by in-house extract system

o Drawn from base of exposure chamber

o Filtered locally

o Flow – individually calibrated to a target of 250 and 380 L/minute for 1000 

and 1500 L chambers, respectively

• Airflow Monitoring:

o Magnehelic gauges; calibrated before study start and monitored 

continuously during exposures

o Differential exposure chamber pressure recorded at 30 minute intervals

4. Administration

Exposure concentrations achieved by varying test substance flow rate. 

Daily Procedure:

• Miran Infra-Red gas spectrophotometer calibration check performed

• Extract airflow runs continuously

• Pigs loaded into designated exposure chamber

• Sponsor cylinder flow (Groups 2 and 3) established

• Animals exposed for 6 hours

o One animal in Group 1 (1148M) received shorter exposures for welfare 

reasons on Days 16 and 17 (353 and 268 minutes, respectively) 

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• Exposure system operational parameters monitored continuously

• Chamber temperature, humidity, airflow rate and static pressure documented 

every 30 minutes during exposure

• Generation terminated – test substance cylinder switched off

• Animals held in chambers for minimum 13 minute system clear out period after 

generation terminated

• Animals returned to home cage at end of exposure

• Animals exposed in the same chamber throughout the study

5. Atmosphere Analysis

5.1 Concentration

The atmosphere concentration was determined as follows:

• Atmosphere analysis - IR spectrophotometer

• Instrument and conditions - Wilks Miran 1A-CVF ambient air analyzer

o Wavelength – 3.26 μm (Groups 1 to 3) 

o Pathlength – 13.02 dial setting (Group 1 up to Exposure Day 22)

14.00 dial setting (Group 1 starting Exposure Day 23, due to 

replacement of instrument, and Groups 2 to 3)

• Measurement frequency – a minimum of 4 samples/group/sex/exposure

documented

• Sampling position remained constant (animal breathing zone) 

5.2 Chamber Temperature and Relative Humidity

Monitored using Extech Humidity and Thermometer gauges as follows:

• Frequency – continuous monitoring; documented every 30 minutes

• Sample location – gauges located inside the exposure chamber

• Gauge position remained constant

5.3 Static Pressure

Monitored using H2O Dwyer Magnehelic gauges as follows:

• Frequency – continuous monitoring; documented every 30 minutes

• Gauge location – outside the exposure chamber

5.4 Chamber Oxygen Levels

Chamber oxygen samples collected during habituation period as follows:

• Sample media – ServoFlex MiniMP (5200) Gas Analyzer

• Sample frequency – minimally one male and one female chamber

• Sample position remained constant (H1)

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5.5 Nominal Concentration

Calculated daily as a summation of all test substance exposure chambers in each 

group. 

6. Data, Abbreviations and Calculations

Data not presented here is retained in the study raw data and the HLS Archive. 

To minimize 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. 

6.1 Abbreviations

Conc. = Mean atmosphere concentration (ppm)

IR = Infrared 

PPM = Parts per million

SD = Standard Deviation

A = Airflow 

D = Duration of exposure 

Wt = Weight 

°C = Degrees Celsius

K = Kelvin

L = Liter

N/A = Not Applicable

6.2 Calculations

Each IR spectrophotometer was calibrated using test substance taken directly from 

the Sponsor cylinder. Absorbance measurements acquired by the IR 

spectrophotometer were used to calculate analyzed concentration (ppm) using a

3rd order fit subjected to least squares regression analysis. 

Nominal Conc. (ppm) = 

7. Results and Discussion 

Results are presented in Tables 1 to 3 of this report.

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7.1 Pre-study Oxygen Measurements

Measured oxygen chamber levels were within protocol specifications above 19%

(ranging between 21.1 to 21.2%). No additional oxygen measurements were required 

during the exposure period.

7.2 Atmosphere Concentration 

Overall mean concentrations are presented in Table 1 and summarized below:

Group Test Substance

Atmosphere Concentration (ppm)

Target Achieved

Males Females

1 Air Control 0 0 0 

2 HFO-1234yf gas 5000 5190 5100

3 HFO-1234yf gas 10000 10200 10200

No HFO-1234yf gas was detected in Group 1 samples. Mean atmosphere 

concentrations were 3.8% and 2.0% above target for Group 2 males and females,

respectively; and 2.0% above target for Group 3. 

The mean nominal concentration of 19100 ppm was 7% less than the overall 

combined IR measured concentration of 20600 ppm for Group 2. The mean nominal 

concentration of 39200 ppm was 4% less than the overall combined IR measured 

concentration of 40800 ppm for Group 3. This was considered a reasonable

experimental difference as seen previously at this laboratory.

7.3 Chamber Temperature and Relative Humidity

Overall mean chamber temperature and relative humidity data are presented in 

Tables 2 and 3, and summarized below:

Group Sex Chamber 

Number Temperature

Relative 

Humidity

(°C) (%)

1 Male

101M 21 59

102M 22 54

103M 21 52

Female

101F 23 54

102F 23 51

2 Male

201M 23 58

202M 24 59

Female

201F 24 55

202F 24 53

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Group Sex Chamber 

Number Temperature

Relative 

Humidity

(°C) (%)

3 Male

301M 23 59

302M 23 60

Female

301F 23 56

302F 23 53

Chamber mean overall temperatures were within protocol specifications between 20 to 

270C. For the Group 1 chamber with only one animal (Chamber No. 103M), the daily 

mean temperature was between 19.1 to 19.60C on Exposure Days 18 to 20 - there 

was no evidence of impact on the study from this deviation. 

Relative humidity was within protocol specifications between 30 to 70% on all 

occasions of measurements. Relative humidity for the test substance groups was 

similar to the air control group.

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Figure 1 Schematic of Inhalation Exposure System

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Table 1 Analytical Atmosphere Concentration – Individual, Mean Exposure

and Nominal Values 

Exposure

Day

Determined Concentration (ppm)a

Nominal

(ppm)a

Group 1

Male Chambers Female Chambers

101M 102M 103Mb Mean 101F 102F Mean

1 0 0 - 0 - - - N/A

2 0 0 - 0 0 0 0 N/A

3 0 0 - 0 0 0 0 N/A

4 0 0 - 0 0 0 0 N/A

5 0 0 - 0 0 0 0 N/A

6 0 0 - 0 0 0 0 N/A

7 0 0 - 0 0 0 0 N/A

8 0 0 - 0 0 0 0 N/A

9 0 0 - 0 0 0 0 N/A

10 0 0 - 0 0 0 0 N/A

11 0 0 - 0 0 0 0 N/A

12 0 0 - 0 0 0 0 N/A

13 0 0 - 0 0 0 0 N/A

14 0 0 - 0 0 0 0 N/A

15 0 0 - 0 0 0 0 N/A

16 0 0 - 0 0 0 0 N/A

17 0 0 - 0 0 0 0 N/A

18 0 0 0 0 0 0 0 N/A

19 0 0 0 0 0 0 0 N/A

20 0 0 0 0 0 0 0 N/A

21 0 0 0 0 0 0 0 N/A

22 0 0 0 0 0 0 0 N/A

23 0 0 0 0 0 0 0 N/A

24 0 0 0 0 0 0 0 N/A

25 0 0 0 0 0 0 0 N/A

26 0 0 0 0 0 0 0 N/A

27 0 0 0 0 0 0 0 N/A

28 0 0 0 0 0 0 0 N/A

29 - - - - 0 0 0 N/A

Mean 0 0 0 0 0 0 0 N/A

SD - - - - - - -

a Refer to Section 6 Data, Abbreviations and Calculations for details.

b Animal 1148M was exposed in a separate chamber from Exposure Day 18 onwards.

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Table 1 Analytical Atmosphere Concentration – Individual, Mean Exposure 

and Nominal Values (cont.)

Exposure

Day

Determined Concentration (ppm)a

Nominal

(ppm)a

Group 2 

Male Chambers Female Chambers

201M 202M Mean 201F 202F Mean

1 5400 5470 5440 - - - 9320

2 5460 5510 5490 5200 5040 5120 20000

3 5290 5360 5330 5200 5280 5240 19000

4 5140 5230 5190 5170 5230 5200 20000

5 5130 5290 5210 5110 5230 5170 19500

6 5180 5380 5280 5060 5070 5070 20900

7 5110 6170 5640 5110 4970 5040 24100

8 5290 5050 5170 5020 4970 5000 13900

9 5150 4930 5040 4910 4990 4950 20000

10 5130 5010 5070 4780 5000 4890 21800

11 5120 5230 5180 4790 4870 4830 18100

12 5180 5190 5190 5430 5120 5280 21800

13 5170 5260 5220 5270 5150 5210 19500

14 5180 5190 5190 5110 5080 5100 17600

15 5020 4970 5000 5220 5220 5220 20900

16 5110 4930 5020 5410 5140 5280 19500

17 4940 5150 5050 5290 5070 5180 19500

18 4960 5030 5000 5200 5250 5230 21800

19 5030 5210 5120 5080 5080 5080 25100

20 5280 5220 5250 5470 4860 5170 18600

21 5200 5040 5120 5330 5080 5210 20000

22 5270 5010 5140 4950 4860 4910 14900

23 5210 5260 5240 4780 4990 4890 20900

24 5100 5010 5060 4890 5240 5070 17200

25 5230 5020 5130 5190 5130 5160 26900

26 4910 4920 4920 5350 4920 5140 12100

27 5250 5380 5320 5410 4970 5190 27900

28 5390 5060 5230 5020 5170 5100 15800

29 - - - 4920 5040 4980 6010

Meanb 5170 5200 5190 5130 5070 5100 19100

SD 132.6 253.5 157.8 201.8 122.8 126.1 4680.4

a Refer to Section 6 Data, Abbreviations and Calculations for details.

b Means are rounded to three significant figures.

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Table 1 Analytical Atmosphere Concentration – Individual, Mean Exposure 

and Nominal Values (cont.)

Exposure

Day

Determined Concentration (ppm)a

Nominal

(ppm)a

Group 3 

Male Chambers Female Chambers

301M 302M Mean 301F 302F Mean

1 10300 10600 10500 - - - 19600

2 9670 10000 9840 10200 10700 10500 39600

3 9520 10300 9910 10100 9980 10000 38200

4 10000 10200 10100 10000 9430 9720 39200

5 10100 10300 10200 10200 9970 10100 39600

6 9910 10400 10200 10400 9850 10100 43800

7 10300 10100 10200 10200 9280 9740 39200

8 9850 10500 10200 10100 9750 9930 39600

9 9690 9910 9800 10100 9520 9810 40600

10 10300 9910 10100 10100 10100 10100 41500

11 10800 9950 10400 10200 10800 10500 42900

12 10600 10000 10300 10300 10200 10300 41000

13 10400 10100 10300 10100 10300 10200 40600

14 10400 9850 10100 10100 9820 10000 38700

15 10600 9580 10100 10500 10100 10300 40100

16 10800 10200 10500 10100 10000 10100 42000

17 10500 9890 10200 10400 9470 9900 41000

18 10700 10000 10400 10100 10200 10200 41500

19 10200 9920 10100 10900 10700 10800 42000

20 10400 9910 10200 10300 10400 10400 41000

21 10300 9830 10100 10900 9770 10300 41000

22 10500 9420 10000 10600 10200 10400 42000

23 10300 9720 10000 10400 10400 10400 40600

24 10200 10100 10200 10900 10800 10900 41500

25 10500 9940 10200 10700 10200 10500 40600

26 10300 9950 10100 9700 9930 9800 41000

27 10600 9730 10200 10300 10700 10500 38700

28 10600 9390 10000 10100 9820 10000 40100

29 - - - 10300 10300 10300 19700

Meanb 10300 9990 10200 10300 10100 10200 39200

SD 336.2 285.2 171.8 288.7 418.8 301.0 5562.3

a Refer to Section 6 Data, Abbreviations and Calculations for details.

b Means are rounded to three significant figures.

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Table 2 Chamber Temperature - Mean Values

Exposure

Day

Temperature (°C)a 

Group 1

Male Chambers Female Chambers

101M 102M 103Mb 101F 102F

1 20.8 22.9 - - -

2 20.2 22.9 - 23.4 23.2

3 20.8 23.2 - 23.0 23.2

4 21.1 23.2 - 23.7 23.5

5 20.8 22.4 - 22.9 22.9

6 21.4 22.8 - 23.4 23.2

7 20.4 22.3 - 22.6 22.8

8 20.7 21.9 - 22.6 22.4

9 20.6 22.1 - 22.9 22.6

10 20.5 21.8 - 22.5 22.3

11 19.9 21.5 - 22.7 22.0

12 20.8 22.2 - 22.8 22.4

13 20.2 22.1 - 22.4 22.4

14 19.9 22.4 - 22.9 22.6

15 20.9 22.6 - 23.0 22.5

16 21.0 22.7 - 22.4 22.7

17 20.6 22.4 - 23.2 22.7

18 20.2 22.5 19.6 23.2 22.4

19 20.0 22.2 19.1 22.5 22.3

20 20.2 22.9 19.5 23.1 22.3

21 21.3 23.1 20.8 24.0 23.5

22 22.0 22.8 21.8 23.7 22.8

23 21.6 22.3 20.5 22.8 22.1

24 21.5 22.2 21.2 22.0 22.0

25 21.3 21.9 21.1 22.7 22.1

26 21.2 21.8 21.0 22.6 22.1

27 22.0 22.8 21.0 23.4 22.4

28 21.3 22.6 20.6 22.3 22.5

29 - - - 22.7 22.0

Mean 20.8 22.4 20.6 22.9 22.6

SD 0.59 0.45 0.83 0.47 0.44

a Refer to Section 6 Data, Abbreviations and Calculations for details.

b Animal 1148M was exposed in a separate chamber from Exposure Day 18 onwards.

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Table 2 Chamber Temperature - Mean Values (cont.)

Exposure

Day

Temperature (°C)a 

Group 2 

Male Chambers Female Chambers

201M 202M 201F 202F

1 23.0 23.9 - -

2 23.6 24.1 24.3 24.0

3 23.9 24.4 24.2 24.2

4 24.3 24.8 25.4 24.8

5 23.9 24.5 24.7 24.6

6 24.0 24.7 24.7 24.4

7 22.6 23.6 23.2 23.4

8 22.6 23.6 23.9 23.5

9 23.1 23.5 24.2 23.9

10 23.2 23.8 23.7 23.2

11 23.0 23.5 23.7 23.7

12 22.8 23.9 23.9 23.8

13 22.7 23.7 24.1 23.5

14 23.3 23.7 24.3 24.1

15 23.0 23.7 24.0 23.9

16 22.6 24.1 24.4 24.4

17 22.1 23.5 24.4 24.6

18 21.6 23.5 23.8 24.5

19 21.4 23.5 23.7 24.2

20 21.5 23.5 24.2 24.4

21 22.1 23.9 24.7 25.0

22 22.0 23.8 23.7 24.3

23 22.1 23.4 23.4 23.5

24 21.4 22.9 23.3 23.7

25 21.1 22.5 23.3 23.1

26 21.8 23.1 23.0 23.4

27 22.1 22.9 23.2 23.3

28 22.5 24.0 24.7 24.5

29 - - 23.1 23.9

Mean 22.6 23.7 24.0 24.0

SD 0.86 0.52 0.59 0.52

a Refer to Section 6 Data, Abbreviations and Calculations for details.

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Table 2 Chamber Temperature - Mean Values (cont.)

Exposure

Day

Temperature (°C)a 

Group 3 

Male Chambers Female Chambers

301M 302M 301F 302F

1 23.4 23.9 - -

2 22.5 23.1 22.9 23.3

3 23.8 24.0 23.3 23.9

4 24.1 24.5 23.9 24.9

5 23.8 23.9 23.4 24.3

6 23.9 24.1 23.8 24.6

7 22.0 22.3 22.0 22.8

8 22.5 22.5 21.7 22.9

9 22.4 23.0 22.3 22.9

10 22.9 22.7 22.2 23.3

11 22.2 22.2 22.0 23.3

12 22.4 22.9 22.8 22.8

13 22.8 23.1 22.5 23.3

14 23.2 22.2 22.5 23.7

15 23.6 22.9 22.8 24.1

16 24.2 23.3 23.7 24.5

17 23.4 23.2 23.0 24.0

18 23.2 22.8 22.5 23.4

19 21.9 21.6 22.5 23.2

20 23.6 23.0 23.0 23.5

21 23.5 22.7 23.1 23.2

22 23.1 22.7 22.6 23.3

23 22.2 22.0 21.4 22.4

24 22.5 22.2 22.1 22.8

25 22.5 22.6 22.1 22.5

26 22.6 22.3 22.1 22.8

27 22.6 22.1 21.6 22.8

28 22.9 22.3 22.7 23.1

29 - - 22.1 22.9

Mean 23.0 22.9 22.6 23.4

SD 0.66 0.71 0.65 0.65

a Refer to Section 6 Data, Abbreviations and Calculations for details.

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Table 3 Chamber Relative Humidity - Mean Values

Exposure

Day

Relative Humidity (%)a

Group 1

Male Chambers Female Chambers

101M 102M 103Mb 101F 102F

1 64 56 - - -

2 63 56 - 55 51

3 67 60 - 57 54

4 66 59 - 59 53

5 65 60 - 60 55

6 68 62 - 61 61

7 58 53 - 53 49

8 57 53 - 52 49

9 62 55 - 56 52

10 65 58 - 58 54

11 57 54 - 52 50

12 60 56 - 55 52

13 63 57 - 57 52

14 59 54 - 53 51

15 63 57 - 56 54

16 66 60 - 60 55

17 57 51 - 50 48

18 55 51 55 50 48

19 53 48 52 48 45

20 53 48 51 47 46

21 56 52 54 52 50

22 51 52 49 52 48

23 51 51 51 49 46

24 52 54 50 54 50

25 52 53 49 55 50

26 52 54 52 54 52

27 52 50 52 50 48

28 53 51 52 49 47

29 - - - 53 50

Mean 59 54 52 54 51

SD 5.7 3.7 1.9 3.8 3.4

a Refer to Section 6 Data, Abbreviations and Calculations for details.

b Animal 1148M was exposed in a separate chamber from Exposure Day 18 onwards.

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Table 3 Chamber Relative Humidity - Mean Values (cont.)

Exposure

Day

Relative Humidity (%)a

Group 2

Male Chambers Female Chambers

201M 202M 201F 202F 

1 60 64 - -

2 60 62 58 55

3 61 63 60 59

4 59 62 57 56

5 62 61 60 59

6 61 64 60 59

7 57 60 54 54

8 56 61 53 52

9 58 59 54 54

10 61 63 59 59

11 53 57 51 52

12 58 60 56 55

13 60 61 56 54

14 57 60 56 54

15 60 61 59 57

16 63 62 60 58

17 55 58 53 51

18 58 59 54 51

19 53 55 50 50

20 61 59 56 51

21 62 62 54 52

22 54 53 52 51

23 48 50 45 46

24 55 55 50 49

25 59 60 54 50

26 57 60 52 52

27 51 52 49 48

28 51 52 50 46

29 - - 58 53

Mean 58 59 55 53

SD 3.8 3.8 3.9 3.8

a Refer to Section 6 Data, Abbreviations and Calculations for details.

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Table 3 Chamber Relative Humidity - Mean Values (cont.)

Exposure

Day

Relative Humidity (%)a

Group 3

Male Chambers Female Chambers

301M 302M 301F 302F

1 62 65 - -

2 65 65 60 58

3 63 66 62 58

4 62 63 59 54

5 61 63 60 57

6 63 64 61 58

7 59 61 57 55

8 60 60 57 55

9 61 63 58 56

10 64 65 62 57

11 60 59 56 52

12 63 65 57 57

13 61 62 58 55

14 59 61 58 55

15 61 63 61 57

16 62 64 60 58

17 58 59 55 53

18 56 58 55 52

19 56 58 52 52

20 56 60 55 54

21 62 62 57 56

22 52 56 52 49

23 50 51 46 45

24 52 54 49 46

25 55 55 51 49

26 56 60 53 49

27 50 52 48 46

28 52 52 48 47

29 - - 57 55

Mean 59 60 56 53

SD 4.4 4.3 4.5 4.1

a Refer to Section 6 Data, Abbreviations and Calculations for details.

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Annex 3 Clinical pathology report

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Annex 4 Anatomic pathology report – organ weights

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Annex 5 Anatomic pathology report – macroscopic and histopathology

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Final Pathology Report 

HLS Study Number 13-6434 

2 1/17/2014 

TABLE OF CONTENTS 

1.0 Executive Summary ......................................................................................................... 3

2.0 Introduction ....................................................................................................................... 4

2.1 Protocol .......................................................................................................................... 4

2.2 Objective ........................................................................................................................ 4

3.0 Methods ............................................................................................................................. 4

3.1 Compliance Statement ................................................................................................... 4

3.2 Study Design .................................................................................................................. 4

Table A. Study Design .......................................................................................................... 4

3.3 Necropsy ........................................................................................................................ 4

3.4 Histological Processing .................................................................................................. 5

4.0 Results ............................................................................................................................... 5

4.1 Animal Mortality .............................................................................................................. 5

4.2 Macroscopic Findings .................................................................................................... 5

4.3 Microscopic Evaluation .................................................................................................. 6

5.0 Conclusions ...................................................................................................................... 6

REFERENCES ............................................................................................................................ 7

Appendix A. Histopathology Incidence Tables .......................................................................... 8

Appendix B. Peer Review Statement ......................................................................................... 74

Appendix C. Quality Assurance Statement .............................................................................. 76

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Final Pathology Report 

HLS Study Number 13-6434 

3 1/17/2014 

1.0 Executive Summary 

The purpose of the study was to assess the toxicity (including cardiotoxicity and/or skeletal 

muscle toxicity) of the test substance when administered via whole-body inhalation to minipigs for 

28 days. 

Methods: According to the protocol, 48 minipigs (24 males and 24 females) were assigned to 

three treatment groups in this study. Animals were exposed to clean air or the test substance by 

inhalation via whole body exposure for 6 hours per day for 7 days per week for up to 28 days. On 

Day 29, all surviving animals were euthanized and subjected to a complete gross necropsy at 

termination. Unscheduled deaths were also subjected to necropsy. Protocol specified tissues 

were collected and processed, embedded in paraffin, sectioned and stained with hematoxylin and 

eosin (H&E) at HLS. The resulting slides from all animals including unscheduled deaths were 

forwarded to HSRL and evaluated via light microscopy by at 

HSRL. A peer review was performed by of Experimental 

Pathology Laboratories, Sterling, VA. 

Conclusion: 

The purpose of the study was to assess the toxicity (including cardiotoxicity and/or skeletal 

muscle toxicity) of the test substance when administered via whole-body inhalation to minipigs for 

28 days. 

At Day 29, there were no HFO-1234yf-related microscopic findings involving the heart (left 

ventricle, right ventricle, and septum) and skeletal muscle (rectus femoris, psoas, and soleus 

muscles as well as the diaphragm). Therefore, under the conditions of this study, there was no 

microscopic evidence of cardiotoxicity or skeletal muscle toxicity following whole body inhalation 

of the test substance at up to 10000 ppm for 28 days in male and female Gottingen minipigs.

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Final Pathology Report 

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2.0 Introduction 

2.1 Protocol 

This report presents the results of the microscopic evaluation of protocol-required tissues to 

assess the toxicity of HFO-1234yf when administered via inhalation to minipigs, HLS Study 

Number 13-6434, for All in-life procedures, tissue harvests, and histology were 

performed at HLS under the direction of , Study Director. 

Microscopic evaluation was completed by of HSRL. 

2.2 Objective 

The purpose of the study was to assess the toxicity (including cardiotoxicity and/or skeletal 

muscle toxicity) of the test substance when administered via whole-body inhalation to minipigs for 

28 days. 

3.0 Methods 

3.1 Compliance Statement 

The portions of this study conducted at HSRL were performed according to the Organization for 

Economic Cooperation and Development (OECD) Principles of Good Laboratory Practices 

ENV/MC/CHEM (98)17. An electronic copy of this report in portable document format (PDF) will 

be provided to HLS in addition to the hard copy. The PDF is a representation of the pathology 

report hard copy. However, only the signed hard copy of the pathology report is considered raw 

data. 

3.2 Study Design 

According to the protocol, 48 animals (24 males and 24 females) were divided into three 

treatment groups. The study design is further described in Table A. 

Table A. Study Design 

Group Exposure Levela

Number of Animals 

Total on Study Terminal NecropsyDay 28 

ppm mg/L Male Female Male Female 

1 0 0 8 8 8 8 

2 5000 26 8 8 8 8 

3 10000 50 8 8 8 8 

a Exposures were 6 hours per day for 7 days per week for up to 28 days. Exposure levels were 

expressed as ppm (part per million of test substance in air) and equivalent mg/L (mg of test substance 

per liter of air) of test substance but study results were reported as ppm. 

3.3 Necropsy 

According to the protocol, all surviving animals were euthanized and subjected to a complete 

gross necropsy on Day 29. Unscheduled deaths were also subjected to necropsy. Necropsies 

consisted of an external examination as well as a detailed internal examination. At scheduled 

termination, the following organs (when present) were weighed before fixation (paired organs 

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Final Pathology Report 

HLS Study Number 13-6434 

5 1/17/2014 

were weighed together): brain, heart, kidneys, liver, and lungs (with mainstem bronchi). For all 

animals, the organs listed below were excised and preserved in 10% neutral buffered formalin. 

Brain 

Heart (left and right ventricles and septum)* 

Kidneys 

Liver 

Lungs (with mainstem bronchi) 

Muscle (diaphragm and bilateral psoas, 

bilateral rectus femoris and soleus)* 

Gross lesions 

* For histopathological evaluation 

3.4 Histological Processing 

Protocol required tissues were processed, embedded in paraffin, sectioned and stained with 

hematoxylin and eosin (H&E) at HLS. A total of eight longitudinal sections were sampled from 

the heart. A cross section and a longitudinal section of three sample areas (midbody and towards 

each end) of the left and right rectus femoris were taken. In addition, a cross and longitudinal 

section of the psoas muscle, diaphragm, and soleus muscle were taken. 

All microscopic slides were forwarded to HSRL and evaluated by 

directly into PathData®

 Systems (PDS) at HSRL. Microscopic findings and microscopic grading 

definitions are presented in the Histopathology Incidence Tables portion of this report, where the 

following abbreviations apply due to PDS character limitations: 

AOFT= Animal Organ Finding Table 

HEART (LEFT VENTR.)= Heart (Left Ventricle) 

HEART (L VENT. FREE)= Heart (Left Ventricular Free Wall) 

HEART (RIGHT VENTR.)= Heart (Right Ventricle) 

MUSCLE (L REC FEMOR)= Muscle (Left Rectus Femoris) 

MUSCLE (R REC FEMOR)= Muscle (Right Rectus Femoris) 

MUSCLE (RIGHT SOLEUS= Muscle (Right Soleus) 

A peer review was performed by Experimental Pathology 

Laboratories, Sterling, VA. 

4.0 Results 

4.1 Animal Mortality 

Group 2 male Animal 2147 was euthanized for welfare reasons on Day 16. At necropsy for this 

animal, the Testing Facility documented a prolapse of the rectum; the presence of clear, 

gelatinous material in the colon; and a single thrombus in the left ventricle as well as a single 

thrombus in the right ventricle. There was no evidence to suggest that these findings were 

associated with exposure to the test substance. 

4.2 Macroscopic Findings 

At Day 29, there were a few sporadic macroscopic findings reported by the Testing Facility but 

there were no incidence patterns/trends for these findings to suggest a relationship to test 

substance exposure. 

Macroscopic findings reported included the following: Group 1 male Animal 1145 with diffuse red, 

dark areas of the right cranial lung lobe; Group 1 female Animal 1652 with a few, clear, 10-19 mm 

cysts involving the caudal pole of the right kidney; Group 2 female Animal 2648 with a few, 10-19 

mm, red, dark areas involving the cardiac region of the gastric mucosa; Group 2 female Animal 

2649 with many 2-9 mm dark areas involving all lung lobes; Group 2 female Animal 2651 with 

bilateral dark mandibular lymph nodes; Group 3 male Animal 3148 with increased pericardial 

fluid; Group 3 male Animal 3151 with many small (≤ 1 mm), red, dark areas involving all lung 

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Final Pathology Report 

HLS Study Number 13-6434 

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lobes; Group 3 male Animal 3152 with focal adhesion of the pericardium to the left ventricle and 

many small (≤ 1 mm), red, dark areas involving the mucosa of the lower rectum; Group 3 female 

Animal 3646 with many 2-9 mm dark areas involving all lung lobes; Group 3 female Animal 3648 

with bilateral dark mandibular lymph nodes; and Group 3 female Animal 3651 with many small (≤

1 mm), red, dark areas involving all lung lobes as well as a few, 10-19 mm, red, dark areas 

involving the cardiac region of the gastric mucosa and one, raised, 2-9 mm dark area of the 

gastric mucosa of the pylorus. Histopathological evaluation of these macroscopic findings was 

not required per the protocol. 

4.3 Microscopic Evaluation 

There were no HFO-1234yf-related microscopic findings involving the heart (left ventricle, right 

ventricle, and septum) and skeletal muscle (rectus femoris, psoas, and soleus muscles as well as 

the diaphragm). 

Microscopic observations involving the heart were of sporadic incidence and primarily limited to 

cellular infiltration (mononuclear or mixed cell) involving the epicardium, myocardium, and/or 

endocardium. These observations were of minimal, insignificant intensity and showed no 

predilection as to anatomic site. An additional irrelevant finding included the single occurrence of 

focal pigmented macrophages within the epicardium in a single cardiac specimen. 

For skeletal muscle, microscopic observations included sporadic mononuclear cell infiltration, 

neutrophil infiltration, histiocytic infiltration, cytoplasmic basophilia of myofibers, and myofiber 

degeneration involving the rectus femoris, soleus, psoas, and/or diaphragmatic muscle. These 

findings were of limited intensity as they predominantly occurred with minimal degree. A single 

animal, Group 2 female 2647, had minimal to moderate myofiber atrophy involving the rectus 

femoris (right and left), soleus (right and left), and psoas (right and left) muscles that was 

accompanied by slight myofiber mineralization of the left psoas muscle. As these atrophic 

changes were only observed in a single female at the low dose exposure level, they were 

deemed to be unrelated to HFO-1234yf. 

The aforementioned cardiac and skeletal muscle changes were incidental and many were those 

that can be observed in the age of Gottingen minipigs used in this study (McInnes, 2012; 

Svendson et al, 1998). Overall, these microscopic observations showed no consistent 

patterns/trends involving incidence or intensity to suggest a relationship to test substance 

exposure. 

5.0 Conclusions 

The purpose of the study was to assess the toxicity (including cardiotoxicity and/or skeletal 

muscle toxicity) of the test substance when administered via whole-body inhalation to minipigs for 

28 days. 

At Day 29, there were no HFO-1234yf-related microscopic findings involving the heart (left 

ventricle, right ventricle, and septum) and skeletal muscle (rectus femoris, psoas, and soleus 

muscles as well as the diaphragm). Therefore, under the conditions of this study, there was no 

microscopic evidence of cardiotoxicity or skeletal muscle toxicity following whole body inhalation 

of the test substance at up to 10000 ppm for 28 days in male and female Gottingen minipigs. 

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

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Huntingdon Life Sciences: 13-6434

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Final Pathology Report 

HLS Study Number 13-6434 

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Appendix A. Histopathology Incidence Tables

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Huntingdon Life Sciences: 13-6434

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 PATHOLOGY REPORT 

 PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 EXPLANATION OF CODES AND SYMBOLS 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 CODES AND SYMBOLS USED AT ANIMAL LEVEL:

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 M = Male animal 

 F = Female animal 

 K0 = Terminal sacrifice group 

 + = Intercurrent death/sacrificed moribund 

 +2 = Sacrificed moribund 

 CODES AND SYMBOLS USED AT ORGAN LEVEL:

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ' = Histologic examination not required 

 + = Organ examined, findings present 

 - = Organ examined, no pathologic findings noted (AOFT only) 

 CODES AND SYMBOLS USED AT FINDING LEVEL:

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 GRADE 1 = Minimal / very few / very small 

 GRADE 2 = Slight / few / small 

 GRADE 3 = Moderate / moderate number / moderate size 

 EXPLANATION OF TABLE TEXT(S) USED AT FINDING LEVEL:

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (LEFT VENTRICULAR FREE WALL) 

 - Mix. cell inf.:endo. 

 = Mixed cell infiltration:endocardium 

 - Mono. cell inf.:epi. 

 = Mononuclear cell infiltration:epicardium 

 - Mono. cell inf.:myo. 

 = Mononuclear cell infiltration:myocardium 

 - Pigment 

 = Pigment:macrophage cytoplasm 

 HEART (LEFT VENTRICLE) 

 - Mono. cell inf.:myo. 

 = Mononuclear cell infiltration:myocardium 

 HEART (RIGHT VENTRICLE) 

 - Mono. cell inf.:myo. 

 = Mononuclear cell infiltration:myocardium 

 HEART (SEPTUM) 

 - Mon. cell inf.:endo. 

 = Mononuclear cell infiltration:endocardium 

 - Mono. cell inf.:myo. 

10

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

328
 PATHOLOGY REPORT 

 PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 EXPLANATION OF CODES AND SYMBOLS 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 EXPLANATION OF TABLE TEXT(S) USED AT FINDING LEVEL:

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 = Mononuclear cell infiltration:myocardium 

 MUSCLE (DIAPHRAGM) 

 - Histiocyte infiltr. 

 = Histiocyte infiltration 

 - Mononuc. cell infil. 

 = Mononuclear cell infiltration 

 - Myofiber degen. 

 = Myofiber degeneration 

 MUSCLE (LEFT RECTUS FEMORIS) 

 - Cytopl. basophilia 

 = Cytoplasmic basophilia:myofiber 

 - Histiocytic infiltr. 

 = Histiocytic infiltration 

 - Mononuc. cell infil. 

 = Mononuclear cell infiltration:interstitium 

 - Myofiber degen. 

 = Myofiber degeneration 

 MUSCLE (LEFT PSOAS) 

 - Mineralization 

 = Mineralization:myofiber 

 - Mononuc. cell infil. 

 = Mononuclear cell infiltration:interstitium 

 MUSCLE (RIGHT RECTUS FEMORIS) 

 - Histiocytic infiltr. 

 = Histiocytic infiltration 

 - Mononuc. cell infil. 

 = Mononuclear cell infiltration:interstitium 

 - Myofiber degen. 

 = Myofiber degeneration 

 - Neutrophil infiltr. 

 = Neutrophil infiltration:interstitium 

 MUSCLE (RIGHT PSOAS) 

 - Mononuc. cell infil. 

 = Mononuclear cell infiltration:interstitium 

 - Myofiber degen. 

 = Myofiber degeneration 

 MUSCLE (RIGHT SOLEUS) 

 - Myofiber degen. 

 = Myofiber degeneration 

11

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

329
 PATHOLOGY REPORT 

 SUMMARY TABLES PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 NUMBER OF ANIMALS WITH MICROSCOPIC FINDINGS BY ORGAN/GROUP/SEX 

 STATUS AT NECROPSY: K0, EXCEPT DEATHS 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 SEX : MALE

 DOSE GROUP: 1 2 3 

 NO.ANIMALS: 8 7 8 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (L VENT. FREE) : 8 7 8 

 - Mono. cell inf.:myo.: 1 - - 

 Grade 1: 1 - - 

 - Mono. cell inf.:epi.: - 1 1 

 Grade 1: - 1 1 

 - Pigment : - - 1 

 Grade 1: - - 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (LEFT VENTR.) : 8 7 8 

 - Mono. cell inf.:myo.: 1 - - 

 Grade 1: 1 - - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (DIAPHRAGM) : 8 7 8 

 - Mononuc. cell infil.: 1 - - 

 Grade 1: 1 - - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (R REC FEMOR) : 8 7 8 

 - Neutrophil infiltr. : 1 - - 

 Grade 1: 1 - - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (RIGHT PSOAS) : 8 7 8 

 - Myofiber degen. : - 1 - 

 Grade 1: - 1 - 

 - Mononuc. cell infil.: 1 - - 

 Grade 1: 1 - - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

12

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

330
 PATHOLOGY REPORT 

 SUMMARY TABLES PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 NUMBER OF ANIMALS WITH MICROSCOPIC FINDINGS BY ORGAN/GROUP/SEX 

 STATUS AT NECROPSY: K0, EXCEPT DEATHS 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 SEX : FEMALE

 DOSE GROUP: 1 2 3 

 NO.ANIMALS: 8 8 8 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (L VENT. FREE) : 8 8 8 

 - Mono. cell inf.:epi.: - - 1 

 Grade 1: - - 1 

 - Mix. cell inf.:endo.: 1 - - 

 Grade 1: 1 - - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (LEFT VENTR.) : 8 8 8 

 - Mono. cell inf.:myo.: - - 1 

 Grade 1: - - 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (RIGHT VENTR.) : 8 8 8 

 - Mono. cell inf.:myo.: - - 1 

 Grade 1: - - 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (SEPTUM) : 8 8 8 

 - Mono. cell inf.:myo.: - 1 - 

 Grade 1: - 1 - 

 - Mon. cell inf.:endo.: 1 - - 

 Grade 1: 1 - - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (DIAPHRAGM) : 8 8 8 

 - Myofiber degen. : - 1 - 

 Grade 1: - 1 - 

 - Mononuc. cell infil.: 1 - 1 

 Grade 1: 1 - 1 

 - Histiocyte infiltr. : 1 - - 

 Grade 1: 1 - - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

13

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

331
 PATHOLOGY REPORT 

 SUMMARY TABLES PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 NUMBER OF ANIMALS WITH MICROSCOPIC FINDINGS BY ORGAN/GROUP/SEX 

 STATUS AT NECROPSY: K0, EXCEPT DEATHS 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 SEX : FEMALE

 DOSE GROUP: 1 2 3 

 NO.ANIMALS: 8 8 8 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (L REC FEMOR) : 8 8 8 

 - Myofiber atrophy : - 1 - 

 Grade 3: - 1 - 

 - Myofiber degen. : 1 1 - 

 Grade 1: 1 1 - 

 - Cytopl. basophilia : 1 - 2 

 Grade 1: 1 - 2 

 - Mononuc. cell infil.: - 1 - 

 Grade 1: - 1 - 

 - Histiocytic infiltr.: - 1 - 

 Grade 2: - 1 - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (LEFT PSOAS) : 8 8 8 

 - Myofiber atrophy : - 1 - 

 Grade 3: - 1 - 

 - Mineralization : - 1 - 

 Grade 2: - 1 - 

 - Mononuc. cell infil.: - 1 - 

 Grade 1: - 1 - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (LEFT SOLEUS) : 8 8 8 

 - Myofiber atrophy : - 1 - 

 Grade 1: - 1 - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (R REC FEMOR) : 8 8 8 

 - Myofiber atrophy : - 1 - 

 Grade 2: - 1 - 

 - Myofiber degen. : - - 1 

 Grade 1: - - 1 

 - Mononuc. cell infil.: - - 1 

 Grade 1: - - 1 

 - Histiocytic infiltr.: - - 1 

 Grade 1: - - 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

14

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

332
 PATHOLOGY REPORT 

 SUMMARY TABLES PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 NUMBER OF ANIMALS WITH MICROSCOPIC FINDINGS BY ORGAN/GROUP/SEX 

 STATUS AT NECROPSY: K0, EXCEPT DEATHS 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 SEX : FEMALE

 DOSE GROUP: 1 2 3 

 NO.ANIMALS: 8 8 8 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (RIGHT PSOAS) : 8 8 8 

 - Myofiber atrophy : - 1 - 

 Grade 3: - 1 - 

 - Mononuc. cell infil.: - 2 - 

 Grade 1: - 2 - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 MUSCLE (RIGHT SOLEUS : 8 8 8 

 - Myofiber atrophy : - 1 - 

 Grade 1: - 1 - 

 - Myofiber degen. : - 1 - 

 Grade 1: - 1 - 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

15

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

333
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ANIMAL NUMBER : 

 1145 1146 1147 1148 1149 1150 1151 1152 

 MK0 MK0 MK0 MK0 MK0 MK0 MK0 MK0 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (L VENT. FREE) - - + - - - - - 

 - Mono. cell inf.:myo. . . 1. . . . . . 

 .........................................................................

 HEART (LEFT VENTR.) - - - - - - + - 

 - Mono. cell inf.:myo. . . . . . . 1. . 

 .........................................................................

 HEART (RIGHT VENTR.) - - - - - - - - 

 .........................................................................

 HEART (SEPTUM) - - - - - - - - 

 .........................................................................

 MUSCLE (DIAPHRAGM) - - - - - - - + 

 - Mononuc. cell infil. . . . . . . . 1. 

 .........................................................................

 MUSCLE (L REC FEMOR) - - - - - - - - 

 .........................................................................

 MUSCLE (LEFT PSOAS) - - - - - - - - 

 .........................................................................

 MUSCLE (LEFT SOLEUS) - - - - - - - - 

 .........................................................................

 MUSCLE (R REC FEMOR) - - + - - - - - 

 - Neutrophil infiltr. . . 1. . . . . . 

 .........................................................................

 MUSCLE (RIGHT PSOAS) - - - - - + - - 

 - Mononuc. cell infil. . . . . . 1. . . 

 .........................................................................

 MUSCLE (RIGHT SOLEUS - - - - - - - - 

 .........................................................................

16

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

334
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ANIMAL NUMBER : 

 1645 1646 1647 1648 1649 1650 1651 1652 

 FK0 FK0 FK0 FK0 FK0 FK0 FK0 FK0 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (L VENT. FREE) - + - - - - - - 

 - Mix. cell inf.:endo. . 1. . . . . . . 

 .........................................................................

 HEART (LEFT VENTR.) - - - - - - - - 

 .........................................................................

 HEART (RIGHT VENTR.) - - - - - - - - 

 .........................................................................

 HEART (SEPTUM) - + - - - - - - 

 - Mon. cell inf.:endo. . 1. . . . . . . 

 .........................................................................

 MUSCLE (DIAPHRAGM) - - + - - - + - 

 - Mononuc. cell infil. . . . . . . 1. . 

 - Histiocyte infiltr. . . 1. . . . . . 

 .........................................................................

 MUSCLE (L REC FEMOR) - + + - - - - - 

 - Myofiber degen. . . 1. . . . . . 

 - Cytopl. basophilia . 1. . . . . . . 

 .........................................................................

 MUSCLE (LEFT PSOAS) - - - - - - - - 

 .........................................................................

 MUSCLE (LEFT SOLEUS) - - - - - - - - 

 .........................................................................

 MUSCLE (R REC FEMOR) - - - - - - - - 

 .........................................................................

 MUSCLE (RIGHT PSOAS) - - - - - - - - 

 .........................................................................

 MUSCLE (RIGHT SOLEUS - - - - - - - - 

 .........................................................................

17

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

335
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ANIMAL NUMBER : 

 2145 2146 2147 2149 2150 2151 2152 2153 

 MK0 MK0 MK0+ MK0 MK0 MK0 MK0 MK0 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (L VENT. FREE) - - - - + - - - 

 - Mono. cell inf.:epi. . . . . 1. . . . 

 .........................................................................

 HEART (LEFT VENTR.) - - - - - - - - 

 .........................................................................

 HEART (RIGHT VENTR.) - - - - - - - - 

 .........................................................................

 HEART (SEPTUM) - - - - - - - - 

 .........................................................................

 MUSCLE (DIAPHRAGM) - - - - - - - - 

 .........................................................................

 MUSCLE (L REC FEMOR) - - - - - - - - 

 .........................................................................

 MUSCLE (LEFT PSOAS) - - - - - - - - 

 .........................................................................

 MUSCLE (LEFT SOLEUS) - - - - - - - - 

 .........................................................................

 MUSCLE (R REC FEMOR) - - - - - - - - 

 .........................................................................

 MUSCLE (RIGHT PSOAS) - - - - - - + - 

 - Myofiber degen. . . . . . . 1. . 

 .........................................................................

 MUSCLE (RIGHT SOLEUS - - - - - - - - 

 .........................................................................

18

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

336
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ANIMAL NUMBER : 

 2645 2646 2647 2648 2649 2650 2651 2652 

 FK0 FK0 FK0 FK0 FK0 FK0 FK0 FK0 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (L VENT. FREE) - - - - - - - - 

 .........................................................................

 HEART (LEFT VENTR.) - - - - - - - - 

 .........................................................................

 HEART (RIGHT VENTR.) - - - - - - - - 

 .........................................................................

 HEART (SEPTUM) - - - + - - - - 

 - Mono. cell inf.:myo. . . . 1. . . . . 

 .........................................................................

 MUSCLE (DIAPHRAGM) - - + - - - - - 

 - Myofiber degen. . . 1. . . . . . 

 .........................................................................

 MUSCLE (L REC FEMOR) - - + - - - - - 

 - Myofiber atrophy . . 3. . . . . . 

 - Myofiber degen. . . 1. . . . . . 

 - Mononuc. cell infil. . . 1. . . . . . 

 - Histiocytic infiltr. . . 2. . . . . . 

 .........................................................................

 MUSCLE (LEFT PSOAS) - - + - - - - - 

 - Myofiber atrophy . . 3. . . . . . 

 - Mineralization . . 2. . . . . . 

 - Mononuc. cell infil. . . 1. . . . . . 

 .........................................................................

 MUSCLE (LEFT SOLEUS) - - + - - - - - 

 - Myofiber atrophy . . 1. . . . . . 

 .........................................................................

 MUSCLE (R REC FEMOR) - - + - - - - - 

 - Myofiber atrophy . . 2. . . . . . 

 .........................................................................

 MUSCLE (RIGHT PSOAS) + - + - - - - - 

 - Myofiber atrophy . . 3. . . . . . 

 - Mononuc. cell infil. 1. . 1. . . . . . 

 .........................................................................

 MUSCLE (RIGHT SOLEUS - - + - - - - - 

 - Myofiber atrophy . . 1. . . . . . 

 - Myofiber degen. . . 1. . . . . . 

 .........................................................................

19

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

337
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ANIMAL NUMBER : 

 3145 3146 3147 3148 3149 3150 3151 3152 

 MK0 MK0 MK0 MK0 MK0 MK0 MK0 MK0 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (L VENT. FREE) - - - - - - - + 

 - Mono. cell inf.:epi. . . . . . . . 1. 

 - Pigment . . . . . . . 1. 

 .........................................................................

 HEART (LEFT VENTR.) - - - - - - - - 

 .........................................................................

 HEART (RIGHT VENTR.) - - - - - - - - 

 .........................................................................

 HEART (SEPTUM) - - - - - - - - 

 .........................................................................

 MUSCLE (DIAPHRAGM) - - - - - - - - 

 .........................................................................

 MUSCLE (L REC FEMOR) - - - - - - - - 

 .........................................................................

 MUSCLE (LEFT PSOAS) - - - - - - - - 

 .........................................................................

 MUSCLE (LEFT SOLEUS) - - - - - - - - 

 .........................................................................

 MUSCLE (R REC FEMOR) - - - - - - - - 

 .........................................................................

 MUSCLE (RIGHT PSOAS) - - - - - - - - 

 .........................................................................

 MUSCLE (RIGHT SOLEUS - - - - - - - - 

 .........................................................................

20

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

338
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TABLE OF INDIVIDUAL MICROSCOPIC FINDINGS (AOFT) 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ANIMAL NUMBER : 

 3645 3646 3647 3648 3649 3650 3651 3652 

 FK0 FK0 FK0 FK0 FK0 FK0 FK0 FK0 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 HEART (L VENT. FREE) - - - - + - - - 

 - Mono. cell inf.:epi. . . . . 1. . . . 

 .........................................................................

 HEART (LEFT VENTR.) - - - + - - - - 

 - Mono. cell inf.:myo. . . . 1. . . . . 

 .........................................................................

 HEART (RIGHT VENTR.) - - - - - + - - 

 - Mono. cell inf.:myo. . . . . . 1. . . 

 .........................................................................

 HEART (SEPTUM) - - - - - - - - 

 .........................................................................

 MUSCLE (DIAPHRAGM) + - - - - - - - 

 - Mononuc. cell infil. 1. . . . . . . . 

 .........................................................................

 MUSCLE (L REC FEMOR) - - - - + - + - 

 - Cytopl. basophilia . . . . 1. . 1. . 

 .........................................................................

 MUSCLE (LEFT PSOAS) - - - - - - - - 

 .........................................................................

 MUSCLE (LEFT SOLEUS) - - - - - - - - 

 .........................................................................

 MUSCLE (R REC FEMOR) - - - + - + - - 

 - Myofiber degen. . . . 1. . . . . 

 - Mononuc. cell infil. . . . . . 1. . . 

 - Histiocytic infiltr. . . . . . 1. . . 

 .........................................................................

 MUSCLE (RIGHT PSOAS) - - - - - - - - 

 .........................................................................

 MUSCLE (RIGHT SOLEUS - - - - - - - - 

 .........................................................................

21

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

339
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ANIMAL HEADING DATA 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 

 ANIMAL SEX DEFINED AND FINAL TEST FIRST AND LAST DATE OF 

 NUMBER M/F STATE OF NECROPSY DAYS DAY UNDER TEST NECROPSY

 .........................................................................

 1145 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 1146 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 1147 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 1148 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 1149 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 1150 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 1151 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 1152 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 .........................................................................

 1645 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 1646 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 1647 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 1648 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 1649 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 1650 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 1651 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 1652 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 .........................................................................

22

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

340
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1145 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

23

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

341
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1146 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

24

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

342
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1147 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 HEART (LEFT VENTRICULAR FREE WALL):

 -Mononuclear cell infiltration:myocardium, focal, grade 1 

 MUSCLE (RIGHT RECTUS FEMORIS):

 -Neutrophil infiltration:interstitium, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

25

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

343
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1148 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

26

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

344
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1149 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

27

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

345
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1150 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 MUSCLE (RIGHT PSOAS):

 -Mononuclear cell infiltration:interstitium, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

28

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

346
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1151 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 HEART (LEFT VENTRICLE):

 -Mononuclear cell infiltration:myocardium, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

29

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

347
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1152 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 MUSCLE (DIAPHRAGM):

 -Mononuclear cell infiltration, mesothelial surface, focal,

 grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

30

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

348
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1645 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

31

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

349
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1646 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 HEART (LEFT VENTRICULAR FREE WALL):

 -Mixed cell infiltration:endocardium, focal, grade 1 

 HEART (SEPTUM):

 -Mononuclear cell infiltration:endocardium, focal, grade 1 

 MUSCLE (LEFT RECTUS FEMORIS):

 -Cytoplasmic basophilia:myofiber, multifocal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

32

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

350
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1647 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 MUSCLE (DIAPHRAGM):

 -Histiocyte infiltration, multifocal, grade 1 

 MUSCLE (LEFT RECTUS FEMORIS):

 -Myofiber degeneration, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

33

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

351
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1648 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

34

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

352
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1649 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

35

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

353
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1650 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

36

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

354
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1651 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 MUSCLE (DIAPHRAGM):

 -Mononuclear cell infiltration, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

37

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

355
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 1, Group 1 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 1652 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

38

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

356
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ANIMAL HEADING DATA 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 

 ANIMAL SEX DEFINED AND FINAL TEST FIRST AND LAST DATE OF 

 NUMBER M/F STATE OF NECROPSY DAYS DAY UNDER TEST NECROPSY

 .........................................................................

 2145 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 2146 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 2147 M K0 +2 16 02-OCT-13 17-OCT-13 17-OCT-13

 2149 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 2150 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 2151 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 2152 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 2153 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 .........................................................................

 2645 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 2646 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 2647 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 2648 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 2649 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 2650 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 2651 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 2652 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 .........................................................................

39

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

357
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2145 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

40

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

358
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2146 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

41

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

359
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2147 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 17-Oct-13 

 DAYS ON TEST : 16 

 DATE OF NECROPSY : 17-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : SACRIFICED MORIBUND 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

42

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

360
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2149 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

43

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

361
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2150 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 HEART (LEFT VENTRICULAR FREE WALL):

 -Mononuclear cell infiltration:epicardium, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

44

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

362
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2151 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

45

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

363
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2152 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 MUSCLE (RIGHT PSOAS):

 -Myofiber degeneration, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

46

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

364
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2153 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

47

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

365
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2645 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 MUSCLE (RIGHT PSOAS):

 -Mononuclear cell infiltration:interstitium, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

48

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

366
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2646 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

49

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

367
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2647 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 MUSCLE (DIAPHRAGM):

 -Myofiber degeneration, focal, grade 1 

 MUSCLE (LEFT RECTUS FEMORIS):

 -Myofiber atrophy, multifocal, grade 3 

 -Myofiber degeneration, focal, grade 1 

 -Mononuclear cell infiltration:interstitium, multifocal,

 grade 1 

 -Histiocytic infiltration, multifocal, grade 2 

 MUSCLE (LEFT PSOAS):

 -Myofiber atrophy, multifocal, grade 3 

 -Mineralization:myofiber, multifocal, grade 2 

 -Mononuclear cell infiltration:interstitium, multifocal,

 grade 1 

 MUSCLE (LEFT SOLEUS):

 -Myofiber atrophy, multifocal, grade 1 

 MUSCLE (RIGHT RECTUS FEMORIS):

 -Myofiber atrophy, multifocal, grade 2 

 MUSCLE (RIGHT PSOAS):

 -Myofiber atrophy, multifocal, grade 3 

 -Mononuclear cell infiltration:interstitium, multifocal,

 grade 1 

 MUSCLE (RIGHT SOLEUS):

 -Myofiber atrophy, multifocal, grade 1 

 -Myofiber degeneration, focal, grade 1 

50

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

368
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 CONT./FF. ANIMAL NO. : 2647

 .........................................................................

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

51

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

369
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2648 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 HEART (SEPTUM):

 -Mononuclear cell infiltration:myocardium, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

52

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

370
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2649 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

53

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

371
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2650 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

54

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

372
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2651 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

55

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

373
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 2, Group 2 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 2652 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

56

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

374
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 ANIMAL HEADING DATA 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 

 ANIMAL SEX DEFINED AND FINAL TEST FIRST AND LAST DATE OF 

 NUMBER M/F STATE OF NECROPSY DAYS DAY UNDER TEST NECROPSY

 .........................................................................

 3145 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 3146 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 3147 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 3148 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 3149 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 3150 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 3151 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 3152 M K0 K0 29 02-OCT-13 30-OCT-13 30-OCT-13

 .........................................................................

 3645 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 3646 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 3647 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 3648 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 3649 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 3650 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 3651 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 3652 F K0 K0 29 03-OCT-13 31-OCT-13 31-OCT-13

 .........................................................................

57

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

375
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3145 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

58

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

376
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3146 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

59

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

377
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3147 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

60

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

378
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3148 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

61

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

379
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3149 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

62

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

380
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3150 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

63

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

381
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3151 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

64

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

382
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3152 SEX : MALE 

 FIRST DAY ON TEST : 02-Oct-13 

 LAST DAY ON TEST : 30-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 30-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 HEART (LEFT VENTRICULAR FREE WALL):

 -Mononuclear cell infiltration:epicardium, multifocal, grade 1 

 -Pigment:macrophage cytoplasm, epicardium, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

65

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

383
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3645 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 MUSCLE (DIAPHRAGM):

 -Mononuclear cell infiltration, interstitium, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

66

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

384
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3646 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

67

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

385
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3647 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

68

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

386
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3648 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 HEART (LEFT VENTRICLE):

 -Mononuclear cell infiltration:myocardium, focal, grade 1 

 MUSCLE (RIGHT RECTUS FEMORIS):

 -Myofiber degeneration, multifocal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

69

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

387
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3649 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 HEART (LEFT VENTRICULAR FREE WALL):

 -Mononuclear cell infiltration:epicardium, focal, grade 1 

 MUSCLE (LEFT RECTUS FEMORIS):

 -Cytoplasmic basophilia:myofiber, multifocal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

70

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

388
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3650 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 HEART (RIGHT VENTRICLE):

 -Mononuclear cell infiltration:myocardium, focal, grade 1 

 MUSCLE (RIGHT RECTUS FEMORIS):

 -Mononuclear cell infiltration:interstitium, focal, grade 1 

 -Histiocytic infiltration, focal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

71

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

389
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3651 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 MUSCLE (LEFT RECTUS FEMORIS):

 -Cytoplasmic basophilia:myofiber, multifocal, grade 1 

 ALL OTHER PROTOCOL TISSUES WITHOUT PATHOLOGIC FINDINGS.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

72

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

390
 PATHOLOGY REPORT 

 INDIVIDUAL ANIMAL DATA PROJECT :13-6434 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEST ITEM : HFO-1234yf PATHOL. NO.: 13077 DSG

 TEST SYSTEM : MINIPIG, 28 Day, Inhalation DATE : 17-JAN-14

 SPONSOR : PATHDATA SYSTEM V6.2c2

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 TEXT OF GROSS AND MICROSCOPIC FINDINGS 

 DOSE GROUP : 3, Group 3 

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

 * ANIMAL NUMBER : 3652 SEX : FEMALE 

 FIRST DAY ON TEST : 03-Oct-13 

 LAST DAY ON TEST : 31-Oct-13 

 DAYS ON TEST : 29 

 DATE OF NECROPSY : 31-Oct-13 

 DEFINED SACR.GROUP : TERMINAL SACRIFICE GROUP 

 STATUS AT NECROPSY : TERMINAL SACRIFICE GROUP 

 .........................................................................

 * NECROPSY FINDINGS

 NO NECROPSY OBSERVATIONS NOTED.

 * MICROSCOPIC FINDINGS

 NO MICROSCOPIC FINDINGS NOTED.

 ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ

73

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

391
Final Pathology Report 

HLS Study Number 13-6434 

74 1/17/2014 

Appendix B. Peer Review Statement

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

392
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

393
Final Pathology Report 

HLS Study Number 13-6434 

76 1/17/2014 

Appendix C. Quality Assurance Statement 

Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

394
Sponsor reference number: 

Huntingdon Life Sciences: 13-6434

395