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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:
From 30 October to 21 November 1991
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1992

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study was performed before LLNA was formally validated and available as OECD guideline.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Lot/batch No.: S1943401
- Storage condition of test material: room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: 70% acetone; 30% polyethylene glycol 400
Concentration / amount:
25 % test substance in vehicle for challenge
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: 70% acetone; 30% polyethylene glycol 400
Concentration / amount:
25 % test substance in vehicle for challenge
No. of animals per dose:
10 male/ 10 female
Details on study design:
RANGE FINDING TESTS:
Several concentrations of the test substance (in 0.01 % dodecylbenzene sulphonate/ 0.9 % physiological saline to 100 %) were injected intradermally to determine a suitable concentration of test substance for induction of sensitisation. Preliminary occluded patch irritation tests were carried out using several concentrations of the test substance (in 70 % acetone/ 30 % polyethylene glycol 400) to determine suitable concentrations of test substance for both induction of sensitisation and for sensitisation challenge. Intradermal injection of 1.0 % test item was the highest concentration inducing mild to moderate signs of skin irritation. The topical application of 99 % test item caused mild erythema. 25 % was the highest non-irritating test item concentration for topical treatment.

MAIN STUDY
A. INDUCTION EXPOSURE
Induction consisted of two stages, intradermal injection followed seven days later by occluded patch application.

Intradermal injection:
The hair was clipped from a 2cm x 4cm area of skin in the dorsal shoulder area and three pairs of intradermal injections made with clipped area as follows:
1. Two 0.1 ml injections of 50 % Freund´s Complete Adjuvant (FCA) in 0.01 % dodecylbenzene sulphonate/ 0.9% physiological saline to 100%.
2. Two 0.1 ml injections of 1.0 % active ingredient Florosa in 0.01 % dodecylbenzene sulphonate/ 0.9 % physiological saline to 100 %.
3. Two 0.1 ml injections of test substance in 0.01 % dodecylbenzene sulphonate/ 0.9 % physiological saline to 100 % mixed 50: 50 with FCA such that the final concentration of test material injected was the same as that in 2. above.

Occluded patch application:
Seven days later the same 2cm x 4cm area was clipped and shaved. A 2cm x 4cm filter paper patch, attached by double-sided adhesive tape to a 4cm x 6cm piece of thin polyethene, was saturated with 99 % active ingredient Florosa placed over the shaved site. The patch was held in place for 48 hours by adhesive plaster wrapped around the trunk behind the forelimbs.


B. CHALLENGE EXPOSURE
Twelve days after the application of the induction patch the guinea pigs were challenged on the clipped and shaved flank by an occluded patch. For each animal, an 8 mm diameter filter paper patch in an 11 mm aluminium patch test cup was saturated with 25 % active ingredient Florosa in 70 % acetone/ 30 % polyethylene glycol 400 and the patch applied to the shaved flank. The patch was held in place for 24 hours by adhesive plaster wound around the trunk. The treatment sites were examined for evidence of sensitisation 24 and 48 hours after removal of the patches using the scoring system.
Challenge controls:
At the same time as the test animals were selected, ten other guinea pigs were selected as treated controls. The control guinea pigs were selected so that their body weights were similar to those of the test guinea pigs at that challenge. The guinea pigs received four intradermal injections of 50 % Freund´s Complete Adjuvant in 0.01 % dodecylbenzene sulphonate/ 0.9% physiological saline to 100 %, followed seven days later by a 24 hour occluded patch of test solvent over the injection sites. At the challenge the control guinea pigs were challenged with test material in exactly the same way as the test animals.
Positive control substance(s):
yes
Remarks:
hexyl cinnamic aldehyde and hydroxycitronellal

Results and discussion

Positive control results:
The results obtained with the positive control substances demonstrated the sensitivity and reliability of the experimental technique used.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25 % test substance in vehicle for challenge
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25 % test substance in vehicle for challenge. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25 % test substance in vehicle for challenge
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25 % test substance in vehicle for challenge. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
No evidence of sensitization reactions in any test animals