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Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
November 95-December 95
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: study done under GLP using OECD guidelines
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The current study was done prior to avaliablity of the LLNA guideline
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
thirty-eight male, albino Dunkin Hartley guinea pigs supplied by DavidHall Limited, Burton-on-Trent, Staffordshire, UK were used. At the start of the main study the animals weighted 392 to 450gr and were approximately eight to twelve weeks old. After an acclimatisation period of at lease five days, each animal was selected at random and given a number unique within the study which was written on a small area of clipped rump using a black indelible marker-pen. The animals were housed singly or in pairs in solid-floor polypropylene cages furnished with woodflakes. Free access to mains tap water and foods (Guinea Pig FD1 Diet, Special Diets Services Limited, Witham, Essex, UK) was allowed throghout the study.
Route:
intradermal
Vehicle:
arachis oil
Remarks:
B.P
Concentration / amount:
Intradermal induction: 1%, 5%, 10% and 25% w/v in arachis B.P
Topical induction: 75%, 50%, 25% and 10% w/w in arachis B.P
Topical challenge: 75%, 50%, 25% and 10% w/w in arachis oil B.P
Main study:
induction: Freunds complete adjuvant plus distilled water 1:1
a 10% w/v suspension of he test material in arachis oil B.P
a 10% w/v emulsion of he test material in 1:1 preparation of Freunds complete adjuvant plus distilled water.
induction control:
Freunds complete adjuvant plus distilled water 1:1
arachis oil B.P
a 50% w/v formulation of arachis oil B.P in Freunds complete adjuvant plus distilled water 1:1
Topical: same as test animals except that the vehicle alone was applied to the filter paper.
Challenge:
Freunds complete adjuvant plus distilled water 1:1
a 10% w/v suspension of the test material in arachis oil B.P
a 10% w/v emulsion of the test material in a 1:1 preparation of Freunds adjuvant plus distilled water 1:1


Route:
intradermal
Vehicle:
arachis oil
Remarks:
B.P
Concentration / amount:
Intradermal induction: 1%, 5%, 10% and 25% w/v in arachis B.P
Topical induction: 75%, 50%, 25% and 10% w/w in arachis B.P
Topical challenge: 75%, 50%, 25% and 10% w/w in arachis oil B.P
Main study:
induction: Freunds complete adjuvant plus distilled water 1:1
a 10% w/v suspension of he test material in arachis oil B.P
a 10% w/v emulsion of he test material in 1:1 preparation of Freunds complete adjuvant plus distilled water.
induction control:
Freunds complete adjuvant plus distilled water 1:1
arachis oil B.P
a 50% w/v formulation of arachis oil B.P in Freunds complete adjuvant plus distilled water 1:1
Topical: same as test animals except that the vehicle alone was applied to the filter paper.
Challenge:
Freunds complete adjuvant plus distilled water 1:1
a 10% w/v suspension of the test material in arachis oil B.P
a 10% w/v emulsion of the test material in a 1:1 preparation of Freunds adjuvant plus distilled water 1:1


No. of animals per dose:
Intradermal induction: A total of four animals were used, each reciving four 0.1 ml injections of only one concentration of test material.
Topical induction: Two animals were treated with four preparations of the test material.
Topical Challenge: Four preparations of the test material were applied to the clipped flanks of two animals .
Details on study design:
RANGE FINDING TESTS (intradermal induction):
Concentrations: 1%, 5%, 10% and 25% v/w in arachis oil B.P.
Total of four guinea pigs were used, each animal receiving four 0.1 ml injections of only one oncentration of test material.
The degree of erythema at the injection site was assessed aprox. 24, 48 and 72 hr and 7 days after injection according to Draize scale).
The highest concentration that caused only mild to moderate skin irritation, and which was well tolerated systemically, was selected for the intradermal induction stage of the main study.

MAIN STUDY
A. INDUCTION EXPOSURE
Shortly before treatment on day 0 the hair was removed from an area appox. 40mm x 60mm on the shoulder region of each animal with veterinary clippers. A row of three injections (0.1 each) was made on each side of the mid-line. Approximately 24 and 48 hr 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 topical application of the test material formulation. A filter patch paper loaded with the test material (75% w/w in arachis oil B.P) as a thick, even layer was applied to the prepared skin and held in place with a strip of surgical adhesive tape. covered with an overlapping length of aluminium foil. The patch and foil were further secured with 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 hr. The degree of erythema and oedema was quantified one and 24 hr following removal of the patches.

B. CHALLENGE EXPOSURE
Shortly before treatment on day 21, an area of approx. 50 mm X 70 mm on both flanks of each animal, was clipped free of hair with veterinary clippers
A square filter paper patch loaded with a thick , even layer of test material at the maximum non irritant concentration (50% w/w in arachis oil B.P) was applied to the shorn right flank of each animal and was held in place with a strip of surgical adhesive tape. To ensure that the maximum non-irritant concentration was used at challenge, the test material at a concentration of 25% w/w in arachis oil B.P. was similarly applied to a skin site on the left shorn flank. The patches were occluded with an overlapping length of aluminium foil and secured with a strip of elastic adhesive bandage wound in a double layer around the torso of each animal.
After 24 hr, the dressing was carefully cut using blunt-tipped scissors, removed and discarded. the challenge sites were swabbed with cotton wool soaked in diethyl ether to remove residual material. The position of the treatment sites was identified by using a black indelible marker-pen.
Prior to the 24-hr observation the flanks were clipped using veterinary clippers to remove re-grown hair.
Approximately 24 and 48 hr after challenge dressing removal, the degree of erythema was quantified. Any other reactions were also recorded
Challenge controls:
Induction of the control animals: Intradermal injections were administrated using an identical procedure to that used for the test animals, except that the injections were:
1. Freund's complete adjuvant plus distilled water in the ration 1:1
2. arachis oil B.P.
3. a 50% w/v formulation of arachis oil B.P in Freunds complete adjuvant/distilled water 1:1.
Positive control substance(s):
no
Key result
Reading:
other: blind
Hours after challenge:
24
Group:
test chemical
Dose level:
50% w/w in arachis oil B.P.
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
very light erythema grade 1
Remarks on result:
other: Reading: other: blind. . Hours after challenge: 24.0. Group: test group. Dose level: 50% w/w in arachis oil B.P.. No with. + reactions: 2.0. Total no. in groups: 10.0. Clinical observations: very light erythema grade 1.
Key result
Reading:
other: blind
Hours after challenge:
48
Group:
test chemical
Dose level:
50% w/w in arachis oil B.P.
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: other: blind. . Hours after challenge: 48.0. Group: test group. Dose level: 50% w/w in arachis oil B.P.. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
other: blind
Hours after challenge:
24
Group:
test chemical
Dose level:
25% w/w in arachis oil B.P.
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
veryslight erythema grade 1
Remarks on result:
other: Reading: other: blind. . Hours after challenge: 24.0. Group: test group. Dose level: 25% w/w in arachis oil B.P.. No with. + reactions: 1.0. Total no. in groups: 10.0. Clinical observations: veryslight erythema grade 1.
Reading:
other: blind
Hours after challenge:
48
Group:
test chemical
Dose level:
25% w/w in arachis oil B.P.
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: other: blind. . Hours after challenge: 48.0. Group: test group. Dose level: 25% w/w in arachis oil B.P.. No with. + reactions: 0.0. Total no. in groups: 10.0.

Skin reaction observed after intradermal induction:

Well defined erythema was noted at the intradermal induction sites of all test group animals at the 24 and 48 hr observation.

No skin reactions were noted at the intradermal induction sites of control group animals at the 24 and 48 hr observations.

Control: No skin reactions were observed

Skin reaction observed after topical induction:

Very slight to well defined erythema with or without very slight oedema was noted at the induction sites of all test group animals at the 1 hr observation and in 18 test group animals at the 24 hr observation.

Control: No skin reactions were observed

Bodyweight: body weight of guinea pigs in the test group between day 0 and 24 were comparable to those in the control group.

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test material F-3014 produced a 15% (3/20) sensitisation rate and was classified as a mild sensitiser. The test material did not meet the criteria for classification as a sensitiser according to EU regulations. No risk phrase is required
Executive summary:

Senstisation study was conducted in compliance with OECD 406 (skin sensitisation) and method B6 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 testmaterial for the induction and challenge phases were selected as follows:

Intradermal induction: 10% w/v in arachis oil B.P

Topical Induction: 75% w/w in arachis oil B.P

Topical Challenge: 50% and 25% w/w in arachis oil B.P

The test material F-3014 produced a 15% (3/20) sensitisation rate and was classified as a mild sensitiser. The test material did not meet the criteria for classification as a sensitiser according to EU regulations. No risk phrase is required

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Migrated from Short description of key information:
The test material F-3014 produced a 15% (3/20) sensitisation rate and was considered a mild sensitiser

Justification for classification or non-classification

The test material F-3014 produced a 15% (3/20) sensitisation rate. The test material did not meet the criteria for classification as a sensitiser according to EU regulations. No risk phrase is required

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