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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:
1994-12-8 to 1995-01-08
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

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
Principles of method if other than guideline:
not applicable
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Existing data is available from a guinea pig maximisation test, new in vivo testing has not been conducted for the purposes of REACH registration

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
not specified
Details on test animals and environmental conditions:
At the start of the main study, the animals weighed 339 - 449 g, and were approximately eight to twelve weeks old. After a minimum acclimatisation period of 5 d, each animal was selected at random and given a unique number.
The animals were housed singly or in pairs in solid-floor polypropylene cages furnished with woodflakes. Free access to mains tap water and food (Guinea pig FD1 Diet, Special Diets Services Limited, Witham, Essex, U.K.) was allowed throughout the study.
The animal room was maintained at a temperature of 18-22°C and relative humidity of 41-64 %. The rate of air exchange was approximately 15 changes per h and the lighting was controlled by a time switch to give 12 h light and 12 h darkness.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
arachis oil
Concentration / amount:
Intradermal induction: 5% w/v in arachis oil B.P. and 5% w/v in a mixture of Freund's Complete Adjuvants plus distilled water (1:1)
Topical induction: undiluted as supplied
Topical challenge: undiluted as supplied and 75% v/v in 1:1 ethanol/diethylphthalate
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
arachis oil
Concentration / amount:
Intradermal induction: 5% w/v in arachis oil B.P. and 5% w/v in a mixture of Freund's Complete Adjuvants plus distilled water (1:1)
Topical induction: undiluted as supplied
Topical challenge: undiluted as supplied and 75% v/v in 1:1 ethanol/diethylphthalate
No. of animals per dose:
A group of 30 guinea pigs was used for the main study, twenty test and ten control animals.
Details on study design:
Induction =Shortly before treatment on Day 0, the hair was removed from an area approximately 40 mm x 60 mm on the shoulder region of each animal with veterinary clippers. A row of three injections (0.1 mL each) was made on each side of the mid-line. The injections were:
i) Freund's Complete Adjuvants plus distilled water in the ratio 1:1
ii) a 5% w/v dilution of test material in arachis oil B.P.
iii) a 5% w/v diltuion of test material in a 1:1 preparation of Freund's Complete Adjuvants plus distilled water
Approximately 24 and 48 h after intradermal injection the degree of erythema at the test material injection sites was evaluated.
One week later (day 7), the same area on the shoulder region used previously for intradermal injections was clipped again and treated with a topical application of the undiluted test material. The undiluted test material was applied to saturation on filter paper whih was held in place by a strip of surgical tape and covered with an overlapping lenght of aluminium foil. The patch and foil were further secured by a strip of elastic adhesive bandage wound in a double layer around the torso of each animal. This occlusive dressing was kept in place for 48 h.
The degree of erythema and oedema was quantified one and 24 h following removal of the patches.

Induction of the control animals= intradermal injections were administered using an identical procedure to that used for the test animals, except that the injections were:
i) Freund's Complete Adjuvants plus distilled water in the ratio 1:1
ii) Arachis oil B.P.
iii) 50 % w/v formulation of arachis oil B.P. in a 1:1 mixture of Freund's Complete Adjuvants/ distilled water
The topical applications followed the same procedure as for the test animals except that nothing was applied to the filter paper. Skin reactions were auantified as for the test animals.
Challenge controls:
Shortly before treatment on Day 21, an area, approximately 50 mm x 70 mm on both flanks of each animal, was clipped free of hair with veterinary clippers.
A quantity of the undiluted test material was applied to saturation to the shorn right flank of each animal on a square of filter paper which was held in place by a strip of surgical adhesive tape. To ensure that the maximum non-irritant concentration was used as challenge, the test material (at a concentration of 75 % v/v in 1:1 ethanol/ diethylphthalate) was applied similarly to a skin site on the left shorn flank. The patches were occluded with an overlapping lenght of aluminium foil and secured by a strip of elastic adhesive bandage wound in a double layer around the torso of each animal.
After 24 h, the dressing was carefully cut using blunt-tipped scissors, removed, and discarded. The challenged sites were swabbed with cotton wool soaked in diethyl ether to remove residual material. Approximately 24 and 48 h after challenge dressing removal, the degree of erythema and oedema formation was quantified.
Positive control substance(s):
not specified

Study design: in vivo (LLNA)

Concentration:
not applicable
No. of animals per dose:
not applicable
Details on study design:
not applicable
Statistics:
not applicable

Results and discussion

Positive control results:
no positive controls

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
Undiluted
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No skin reactions
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Undiluted. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No skin reactions.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
Undiluted
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No skin reactions
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: Undiluted. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No skin reactions.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
Undiluted
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No skin reactions
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Undiluted. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No skin reactions.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
Undiluted
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No skin reactions
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: Undiluted. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No skin reactions.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
75 % v/v in 1:1 Ethanol/diethylphthalate
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No skin reactions
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 75 % v/v in 1:1 Ethanol/diethylphthalate. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No skin reactions.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
75 % v/v in 1:1 Ethanol/diethylphthalate
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No skin reactions
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 75 % v/v in 1:1 Ethanol/diethylphthalate. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No skin reactions.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
75 % v/v in 1:1 Ethanol/diethylphthalate
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No skin reactions
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 75 % v/v in 1:1 Ethanol/diethylphthalate. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No skin reactions.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
75 % v/v in 1:1 Ethanol/diethylphthalate
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No skin reactions
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 75 % v/v in 1:1 Ethanol/diethylphthalate. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No skin reactions.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: not applicable
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: not applicable

Any other information on results incl. tables

not applicable

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test material was classified as a non-sensitiser to guinea pig skin. The test material did not meet the criteria for classification as a sensitiser according to EC labelling regulations. No risk phrase is required.
Executive summary:

A guinea pig maximisation test was performed to assess the skin contact sensitisation potential of the test material in albino guinea pigs. The method used followed the one described in the current OECD guidelines no. 406 "Skin sensitisation" and the method B.6 of Commission Directive 92/69/EEC.

Twenty test and ten control animals were used for the main study.

Based on the results of sighting tests, the concentrations of test material for the inducton and challenges phases were selected as follows:

Intradermal induction

5% w/v in arachis oil B.P.

Topical induction

undiluted as supplied

Topical challenge

undiluted as supplied and 75% v/v in 1:1 ethanol/diethylphthalate

The test material produced a 0% (0/20) sensitisation rate and was classified as a non-sensitiser to guinea pig skin. The test material did not meet the criteria for classification as a sensitiser according to EC labelling regulations. No risk phrase is required.