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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07-JAN-2000 to 02-MAY-2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This GLP-compliant study was conducted in accordance with OECD guideline 406 and EU method B.6.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
The Buehler test was conducted for hazard and risk assessmet purposes before REACH entered into force.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Substance type: monoconstituent 
- Physical state: colourless liquid
- Analytical purity: 100%
- Density: 1.64
- Lot/batch No.: 6-99
- Manufacturing date: 24 May 1999
- Expiration date of the lot/batch: May 2001
- Storage condition of test material: at room temperature
Specific details on test material used for the study:
- Name of test material (as cited in study report): Perfluorosulfonyl-vinyl-ether
- Impurities, purity test date: no data available

- Substance type: monoconstituent 
- Physical state: colourless liquid
- Analytical purity: 100%
- Density: 1.64
- Lot/batch No.: 6-99
- Expiration date of the lot/batch: MAY-2001
- Storage condition of test material: at room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Italia S.p.A. / Via Indipendenza, 11 - 23885 Calco (Lecco) / ITALY
- Age at study initiation: ca. 5 weeks at the start of the experiment
- Weight at study initiation: 368 to 421 g at the start of the experiment
- Housing: 2 or 3 animals/cage in an air-conditioned room, in wire cages (40.5x38.5x18h) with a stainless steel feeder
- Diet: ad libitum with a standard diet coded "8 GP 22" (Charles River Italia's feed licensee Mucedola S.r.l., Settimo Milanese) supplemented by the producer with vitamins and trace elements
- Water: ad libitum, filtered water from municipal water main system
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2°C
- Humidity: 55 ± 10%
- Air changes: ca. 15-20 per hr filtered on HEPA 99.97%
- Photoperiod: 12 hrs dark / 12 hrs light (7 a.m. - 7 p.m.)

IN-LIFE DATES: From 07-JAN-2000 (preliminary test) or 28-JAN-2000 (main study) to 02-MAR-2000

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: acetone
Concentration / amount:
75% in acetone
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: acetone
Concentration / amount:
75% in acetone
No. of animals per dose:
20 animals for the treated group
10 animals for the control group
Details on study design:
RANGE FINDING TESTS:
The preliminary test was performed on a total of 2 animals in order to select the highest concentration that causes mild irritation to be used in the induction phase and the highest concentration that proves not to be irritating for the challenge exposure.
Two animals were treated by topical application (in the dorsal region) with 3 occlusive patches (2x3 cm). The patches were loaded with 0.3 mL of three test article concentrations in acetone (25, 50 and 75%).
Twenty four hours after the administration, the patches were removed and the animals observed for up to 48 h for local reactions. Results obtained in the preliminary test are reported in Table 1 below in "any other information on materials and methods incl. tables).
It was decided to assay the possible sensitising activity of the test article with the 75% test article concentration in the induction and in the challenge phases.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 (on days 0, 7 and 14)
- Exposure period: 14 days
- Test groups: group 1 (left flank: test article)
- Control group: group 2 (left flank: vehicle)
- Site: one day prior to the beginning of the induction exposure (day -1), an area of ca. 6x6 cmq on the left flank of the animals of the two groups was clipped. Care was taken to avoid abrading the skin, which could alter the results of the study. On the first day of administration (day 0), a filter paper (3M Whatman 2x3 cm) was fully-loaded with the test article or vehicle and applied to the skin areas clipped the day before. The patch was covered by an overlapping impermeable, hypoallergenic, plastic adhesive tape (3M Blenderm). This in turn was firmly secured by adhesive bandage (3M Micropore), wound around the torso of the animal.
- Frequency of applications: once a week
- Duration: 6 hrs per patch
- Concentrations: 75% (0.3 mL/patch for each exposure)

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Duration of exposure: 6 hours
- Day(s) of challenge: on day 28
- Test groups: group 1 (right posterior flank: test article; right anterior flank: vehicle)
- Control group: group 2 (the animals were challenged in the same way as group 1)
- Site: an area of ca. 6x6 cm on the right flank of the animals of the two groups was clipped. An occlusive patch (2x3 cm) loaded with the test article was applied to the posterior of the right flank of all animals, while the vehicle was similarly applied to the anterior of the flank. The patches were kept in contact for 6 hrs.
- Concentrations: 75% (0.3 mL/patch for each exposure)
- Evaluation: on day 29, 30 hrs after the application of the challenge patch; (approximately 21 hours after removing the patches the challenge areas were clipped and approximately 3 hours later the skin was observed and any reaction recorded (day 29). Twenty-four hours after the above observation a second observation was made and once again recorded (day 30).

OTHER:
- Evaluation of skin reactions:
0: absent
1: discrete or patchy erythema
2: moderate and confluent erythema
3: intense erythema and swelling

The results were expressed in terms of incidence and severity of response:
- incidence: the number of animals showing responses of 1 or greater at either 24 or 48 hours divided by the number of animals tested.
- severity: the sum of the test grades divided by the number of animals tested.
Positive control substance(s):
not required

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
75% test item in acetone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no positive skind reaction. No moratlity occurred. No signs of general toxicity observed in any animal. BW gain was normal.
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
75% test item in acetone
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no positive skind reaction. No moratlity occurred. No signs of general toxicity observed in any animal. BW gain was normal
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle alone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no positive skind reaction. No moratlity occurred. No signs of general toxicity observed in any animal. BW gain was normal
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle alone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no positive skind reaction. No moratlity occurred. No signs of general toxicity observed in any animal. BW gain was normal
Remarks on result:
no indication of skin sensitisation

Any other information on results incl. tables

No mortalities occurred and no signs of general toxicity were observed in any animal. The body weight gain of the animals throughout the test was normal.

Twenty-four hours after each treatment of the induction phase no signs of skin irritation were observed in all test article-treated animals. No reaction was seen after treatment with the vehicle alone.

Table 2 shows the results obtained at the challenge exposure with a 75% concentration of the test article applied with an occlusive patch (vehicle acetone). No animals showed positive reactions at the challenge.

 

Table 2: Challenge in treated animals

 

Challenge

Day 29

Day 30

Group 1

Test article

Vehicle

Test article

Vehicle

Guinea pig No.

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

 

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

No. of positive animals at the challenge: 0

 

No skin reactivity was observed in the negative control group (see Table 3).

 

Table 3: Challenge in control animals

 

Challenge

Day 29

Day 30

Group 2

Test article

Vehicle

Test article

Vehicle

Guinea pig No.

21

22

23

24

25

26

27

28

29

30

 

0

0

0

0

0

0

0

0

0

0

 

0

0

0

0

0

0

0

0

0

0

 

0

0

0

0

0

0

0

0

0

0

 

0

0

0

0

0

0

0

0

0

0

No. of positive animals at the challenge: 0

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the experimental conditions applied, the test article did not show a skin sensitizing capacity.
Executive summary:

The potential of the test material to act as a skin sensitizer was investigated in accordance with the OECD guideline 406 and EU Method B.6 (Buehler test) and in compliance with good laboratory practices.

 

The study assessed the skin sensitisation potential of the test material in male Dunkin-Hartley guinea-pigs following topical application in the dorsal region. The induction phase consisted of guinea-pigs (n = 20) treated three times (once per week, 6 hrs per patch) with 0.3 mL/patch of the test material diluted in acetone at the concentration of 75%. The challenge phase consisted of a single application of the test article at the concentration of 75% (6 hrs per patch, 15 days after the last inductive exposure). A further group of 10 animals was treated with the vehicle alone during the induction phase and in the same way as the test group animals at the challenge.

 

No animals treated with the test article showed positive reaction at the challenge.

 

On the basis of this result, under the experimental conditions applied, the test article did not show a skin sensitizing capacity.