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Working for a better future

HFCPE: Skin Irritation to the Rabbit 

HLS study number: VHJ0003 

Version ID: Final 

Issue date: 28 September 2010 
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Details of Sponsor and Test Facility 

Sponsor ZEON Corporation 

 Shin Marunouchi Center Building 

 1-6-2 Marunouchi 

 Chiyoda-ku 

 Tokyo, 100-8246 

 JAPAN 

 

Test facility Huntingdon Life Sciences 

 Eye Research Centre 

Eye 

Suffolk 

IP23 7PX 

 UK 
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Table of Contents 

Details of Sponsor and Test Facility ....................................................................................... 2

Compliance with Good Laboratory Practice............................................................................ 4

Quality Assurance Statement ................................................................................................. 5

Contributing Scientist.............................................................................................................. 6

Summary................................................................................................................................. 7

1. Introduction .................................................................................................................... 8

2. Test substance............................................................................................................... 9

3. Experimental Procedure............................................................................................... 10

3.1 Animal management ........................................................................................ 10

3.2 Test substance preparation.............................................................................. 10

3.3 Treatment procedure........................................................................................ 10

3.4 Serial observations........................................................................................... 11

3.4.1 Clinical signs......................................................................................... 11

3.4.2 Dermal responses ................................................................................ 11

3.4.3 Interpretation of responses................................................................... 12

3.4.4 Termination........................................................................................... 13

3.5 Archives............................................................................................................ 13

3.6 Deviations from protocol................................................................................... 13

4. Results ......................................................................................................................... 14

4.1 Clinical signs .................................................................................................... 14

4.2 Dermal responses ............................................................................................ 14

5. Conclusion ................................................................................................................... 15

List of Tables 

Table 1 Mean values for erythema and oedema........................................................ 16

Table 2 Individual values for erythema and oedema.................................................. 17

List of Annexes 

Annex 1 Weight of Evidence ....................................................................................... 18

Annex 2 GLP Compliance Statement.......................................................................... 19


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Contributing Scientist 

HFCPE: Skin irritation to the rabbit 

Study management 

P B Rees BSc (Hons) CBiol MSB 

Study Director 
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Summary 

A study was performed to assess the skin irritation potential of HFCPE to the rabbit. The 

method followed was that described in: 

EEC Methods for the determination of toxicity and other health effects. Council 

Regulation No. 440/2008. Part B, Method B.4 Acute Toxicity: Dermal 

irritation/corrosion. 

OECD Guideline for Testing of Chemicals No.404 “Acute Dermal 

Irritation/Corrosion”. Adopted 24 April 2002. 

EPA Health Effects Test Guidelines OPPTS 870.2500 Acute Dermal Irritation 

EPA 712-C-98-196. August 1998. 

Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration 

of Agricultural Chemicals, Skin Irritation (2-1-4), 12 Nohsan No. 8147, Agricultural 

Production Bureau, November 24, 2000. 

Three rabbits received a single four hour, semi-occlusive, dermal administration of 

approximately 0.5 mL of the test substance as supplied and were observed for four days. 

Very slight erythema was apparent one hour after bandage removal in a single animal; no 

other dermal response was apparent at any time during the study. 

The means of scores for these reactions at approximately 24, 48 and 72 hours after 

administration, calculated separately for each animal, are summarised below: 

Means of scores at approximately 24, 48 and 72 hours 

Animal number Erythema Oedema 

 

93 0.0 0.0 

96 0.0 0.0 

97 0.0 0.0 

EC trigger values* ≥2 ≥2 

 

* Classification is triggered if means of scores for either effect are ≥ 2 for 

 two or three animals (or if effects persist to Day 14 in at least two animals). 

The Primary Irritation Index was calculated to be 0.0; HFCPE was classified as ‘non-irritant’ 

according to the criteria of the ECETOC and did not require labelling with the risk phrase 

R38, “Irritating to skin”, in accordance with Commission Directive 2001/59/EC. 
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1. Introduction 

The study was designed to assess skin irritation potential of HFCPE following a single 

dermal application to rabbits. The test substance may come into contact with skin during 

handling or use. 

The study was conducted in compliance with the following guidelines: 

EEC Methods for the determination of toxicity and other health effects. Council 

Regulation No. 440/2008. Part B, Method B.4 Acute Toxicity: Dermal 

irritation/corrosion. 

OECD Guideline for Testing of Chemicals No.404 “Acute Dermal 

Irritation/Corrosion”. Adopted 24 April 2002. 

EPA Health Effects Test Guidelines OPPTS 870.2500 Acute Dermal Irritation 

EPA 712-C-98-196. August 1998. 

Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration 

of Agricultural Chemicals, Skin Irritation (2-1-4), 12 Nohsan No. 8147, Agricultural 

Production Bureau, November 24, 2000. 

The albino rabbit was chosen as it has been shown to be a suitable model for skin irritation 

studies and is the animal recommended in the test guidelines. 

The amount of test substance administered was chosen in compliance with the guidelines. 

The protocol was approved by the Study Director and Huntingdon Life Sciences 

Management on 11 June 2010 and by the Sponsor on 15 June 2010. 

The experimental start date was 18 June 2010 and the experimental completion date was 

29 June 2010. 
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2. Test substance 

Identification: HFCPE 

Intended use: Electronic applications 

Description: Colourless liquid 

Storage conditions: Room temperature 

Batch number: Lot No 20090601 

Date of receipt: 22 June 2009 

Expiry date: End June 2010 

Purity: 99.3% 
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3. Experimental Procedure 

3.1 Animal management 

Animals for this study were selected from a stock supply of healthy adult rabbits of the 

New Zealand White strain. They were in the weight range of 3.65 to 3.93 kg and 

approximately 41 weeks of age, prior to treatment (Day 1). All rabbits were acclimatised to 

the experimental environment for a period of 30 weeks prior to the start of the study. 

The rabbits were housed individually in plastic cages with perforated floors at the 

Eye Research Centre, Eye, Suffolk, IP23 7PX. 

Each rabbit was offered 125 g of a standard laboratory rabbit diet per day; drinking water was 

provided ad libitum. The batch of diet used for the study was analysed for nutrients, possible 

contaminants and micro-organisms likely to be present in the diet and which, if in excess of 

specified amounts, might have an undesirable effect on the test system. The animals were 

given a dietary supplement of hay. 

During the acclimatisation and study period the animals were given small soft white 

untreated wood blocks for environmental enrichment. 

Results of routine physical and chemical examination of drinking water, as conducted by the 

supplier are made available to Huntingdon Life Sciences Limited. 

Animal room environmental controls were set to maintain temperature within the range 16 to 

20°C, and relative humidity within 40 to 70%. These environmental parameters were 

recorded and the permanent record archived with other departmental raw data. Lighting was 

controlled by means of a time switch to give 12 hours of artificial light (06:00 to 18:00 GMT) 

in each 24 hour period. 

Each animal was identified by a numbered tag placed through the edge of one ear. This 

identification was unique within the Department throughout the duration of the study. Each 

cage was identified by a coloured label displaying the study number and animal number. 

3.2 Test substance preparation 

HFCPE was administered as supplied by the Sponsor. 

The absorption of HFCPE was not determined. 

The identity, strength and purity of the test substance received, its stability under the storage 

conditions and the conditions of administration were the responsibility of the Sponsor. 

3.3 Treatment procedure 

On the day before application of the test substance, hair was removed with clippers from the 

dorso-lumbar region of each rabbit exposing an appropriate sized area of skin. 

Approximately 0.5 mL of the test substance was applied under a 2-ply 25 mm x 25 mm 

porous gauze pad secured with ‘blenderm’ surgical tape to intact skin sites on three animals. 
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An additional site was similarly treated with the exception of test substance and acted as a 

control. 

A single animal (number 93) received three exposures of three minutes, one or four hours 

duration in a step-wise manner and acted as a preliminary screen. In the absence of a severe 

effect on removal of the dressings the next exposure was initiated. In the absence of a severe 

response in this animal, two further animals (numbers 96 and 97) were committed to the 

study each receiving a single four hour exposure. 

For exposures of one hour or more each treatment site was covered with cotton wool and 

"Tubigrip" elasticated bandage dressing for the duration of the exposure period. The animals 

were returned to their cages immediately after treatment. 

At the end of the exposure period the semi-occlusive dressing and gauze pad were removed 

and the treatment site was washed with lukewarm water (30-40°C) to remove any residual 

test substance. The treated area was blotted dry with absorbent paper. 

3.4 Serial observations 

3.4.1 Clinical signs 

All animals were observed daily for signs of ill health or toxicity. 

3.4.2 Dermal responses 

Examination of the treated skin was made on removal of the dressings (for 3 minute or 

one hour exposures) and approximately 1, 24, 48 and 72 hours later. Only the data for the 

four hour exposure are reported, the data from the three minute and one hour exposures are 

held in the archives. 

Local dermal irritation was assessed using the prescribed numerical system: 

Erythema and eschar formation: 

No erythema 0 

Very slight erythema (barely perceptible) 1 

Well-defined erythema 2 

Moderate to severe erythema 3 

Severe erythema (beet redness) or eschar formation 

(injuries in depth) preventing grading of erythema 4 

Oedema formation: 

No oedema 0 

Very slight oedema (barely perceptible) 1 

Slight oedema (edges of area well-defined by definite raising) 2 

Moderate oedema (raised approximately 1 millimetre) 3 

Severe oedema (raised more than 1 millimetre and 

extending beyond the area of exposure) 4 
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3.4.3 Interpretation of responses 

Primary Irritation Index 

A primary irritation index (PII) was calculated from the erythema and oedema scores 

according to the following formula as described in Technical Report No. 66 “Skin irritation 

and Corrosion: Reference chemicals data bank” (March 1995) ECETOC, Brussels. 

PII= ∑ Erythema at 24/48/72 hours + ∑ Oedema at 24/48/72 hours

 3 x number of animals 

The maximum possible score was 8.0. The PII was then used to classify the test substance as 

follows: 

Primary Irritation Index Classification 

0 non-irritant 

>0 - 2.0 mildly irritating 

2.1 - 5.0 moderate irritant 

5.1 - 6.0 moderate to severe irritant 

> 6.0 severe irritant 

This classification system is a modification based on “Appraisal of the Safety of Chemicals in 

Foods Drugs and Cosmetics” published by the Association of Food and Drug Officials of the 

United States, 1959. 

EU Classification 

The Official Journal of the European Communities (Directive 2001/59/EC) contains the 

following criteria for classification of irritants and corrosives to skin. 

Substances and preparations shall be classified corrosive if, when applied to healthy intact 

animal skin, full thickness destruction of skin tissue on at least one animal occurs as a result 

of up to four hours exposure. The following risk phases shall be assigned in accordance with 

the following criteria: 

R35: Causes severe burns 

- if, when applied to healthy intact animal skin, full thickness destruction of the skin occurs as 

a result of up to three minutes exposure. 

R34: Causes burns 

- if, when applied to healthy intact animal skin, full thickness destruction of the skin occurs as 

a result of up to four hours exposure. 

Non-corrosive substances and preparations shall be classified irritant if, when applied to 

healthy intact animal skin for up to four hours, significant inflammation is caused and is 

present 24 hours or more after the end of the exposure period. Inflammation is significant if 

the mean value of the scores for either erythema (including eschar formation) or oedema 

formation observed in two or more animals is equivalent to the value of two or more. All 

scores at 24, 48 and 72 hours should be used in calculating the respective mean values. 
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Inflammation of the skin is also significant if it persists in at least two animals at the end of 

the observation period. Particular effects e.g. hyperplasia, scaling, discolouration, fissures, 

scabs and alopecia should be taken into account. 

The following risk phrase shall be assigned: 

R38: Irritating to skin 

3.4.4 Termination 

Following completion of the observation period the animals were humanely killed by an 

intravenous injection of sodium pentobarbital. 

3.5 Archives 

All raw data arising from the performance of this study at Huntingdon Life Sciences is the 

property of the Sponsor and will be lodged together with a copy of the final report in the 

Huntingdon Life Sciences Archive. 

Such records will be retained for a minimum of one year from the date on which the Study 

Director signs the final report. At the end of the retention period the Sponsor will be 

contacted and advice sought on the return, disposal or further retention of the records. 

Huntingdon Life Sciences will retain the Quality Assurance records relevant to this study and 

a copy of the final report in its archive indefinitely. 

3.6 Deviations from protocol 

The following deviation from protocol occurred: 

Section 3.1 (Animal management) of the Study Protocol indicates that the animals to be 

employed on this study would be within the approximate age range of 12 to 40 weeks of age 

on the day of dosing. 

The dates of birth and dose administration for the animals used on the study are given below: 

Animal 

number 

Date of Birth Date of 

administration 

Age at 

administration 

93 9 September 2009 21 June 2010 41 Weeks 

96 8 September 2009 23 June 2010 41 Weeks 

97 8 September 2009 23 June 2010 41 Weeks 

The animals were healthy young adult animals and the use of slightly older than intended 

animals had no impact upon the study. 
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4. Results 

4.1 Clinical signs 

There was no sign of toxicity or ill health in any rabbit during the observation period. 

4.2 Dermal responses 

Very slight erythema was apparent one hour after bandage removal in a single animal; no 

other dermal response was apparent at any time during the study. 
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5. Conclusion 

The Primary Irritation Index was calculated to be 0.0; HFCPE was classified as ‘non-irritant’ 

according to the criteria of the ECETOC and did not require labelling with the risk phrase 

R38, “Irritating to skin”, in accordance with Commission Directive 2001/59/EC. 
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Table 1 Mean values for erythema and oedema 

24, 48 and 72 hours after removal of dressings * 

Animal number Erythema Oedema 

and sex Test Control Test Control 

 

93 M 0.0 0.0 0.0 0.0 

96 M 0.0 0.0 0.0 0.0 

97 M 0.0 0.0 0.0 0.0 

 

* 4 hour exposure 

M Male 
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Table 2 Individual values for erythema and oedema

Test site: 0.5 mL HFCPE } semi occluded for four hours Control site: No treatment } 

Score after removal of dressings 

1 hour 24 hours 48 hours 

Animal number and sex Type of Response Test site Control site Test site Control site Test site Control si

 Erythema 0 0 0 0 0 0 

93 M # 

 Oedema 0 0 0 0 0 0  Erythema 0 0 0 0 0 0 

96 M 

 Oedema 0 0 0 0 0 0  Erythema 1 0 0 0 0 0 

97 M 

 Oedema 0 0 0 0 0 0 

M Male # Sentinel animal 
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Annex 1 Weight of Evidence 

Prior to undertaking in-vivo skin irritation testing the Study Director conducted a weight-ofthe-evidence analysis to ensure that the in-vivo testing was sufficiently justified. The 

following factors have been taken into consideration: 

1. Nature of test substance. 

A colourless liquid for use in electronic applications. The Sponsor indicated that the test 

substance has the following: 

Molecular weight: 194.05 

Chemical name: 1,3,3,4,4,5,5-Heptafluorocyclopentene 

Log Pow: 2.4 

Structure: 

2. Evaluation of existing human and animal data (including dermal toxicity) 

Searches for the test material failed to yield any data. 

Searches for related materials yielded: 

OFCPE (1,2,3,3,4,4,5,5-Octofluorocyclopentane): skin irritation study in rabbits (HLS 

study ZCE037): no dermal irritation. Eye irritation study in rabbits (HLS study ZCE038): 

irritating to the eye 

HFCPA (1,1,2,2,3,3,4-Heptafluorocyclopentane): skin irritation study in rabbits (HLS 

study ZCE034): no dermal irritation. Eye irritation study in rabbits (HLS study ZCE035): 

non-irritating to the eye 

3. Analysis of structure activity relationships (SAR). 

None available. 

4. Physicochemical properties. 

pH (as measured at Huntingdon Life Sciences) – 5.0 

5. Results from in-vitro or ex-vivo tests. 

EPISKIN (HLS study VHJ0002): predicted as non-irritant to the skin 

BCOP (HLS study VHJ0004): no severe eye irritancy or corrosivity prediction 
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Annex 2 GLP Compliance Statement