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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:
1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Selection criteria for dose selection were not clear. reliability was assessed in the Risk Assessment Report under the Existing substances Regulation.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
see: any other information on materials and methods
GLP compliance:
no
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
The test substance was the commercially available grade of HHCB, which is ca. 65% HHCB in DEP

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
Six male and four female Albino Dunkin/Hartley guinea pigs (weight 316-350 g)

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
other: 0.01% dodecylbenzene sulphonate in 0.9% saline (DOBS/saline) for the intradermal induction injection, 70% acetone / 30% polyethylene glycol 400 (aceton/PEG400) for the challenge patch.
Concentration / amount:
Intradermal injection: 0.5% Galaxolide 50 (equivalent to 0.325% HHCB) in vehicle (0.01% dodecylbenzene sulphonate in 0.9% saline (DOBS/saline))
Induction patch: 100% Galaxolide 50 (equivalent to 65% HHCB)
Challenge patch: 25% Galaxolide 50 (equivalent to 16.25% HHCB) in vehicle (70% acetone / 30% polyethylene glycol 400 (aceton/PEG400))
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: 0.01% dodecylbenzene sulphonate in 0.9% saline (DOBS/saline) for the intradermal induction injection, 70% acetone / 30% polyethylene glycol 400 (aceton/PEG400) for the challenge patch.
Concentration / amount:
Intradermal injection: 0.5% Galaxolide 50 (equivalent to 0.325% HHCB) in vehicle (0.01% dodecylbenzene sulphonate in 0.9% saline (DOBS/saline))
Induction patch: 100% Galaxolide 50 (equivalent to 65% HHCB)
Challenge patch: 25% Galaxolide 50 (equivalent to 16.25% HHCB) in vehicle (70% acetone / 30% polyethylene glycol 400 (aceton/PEG400))
No. of animals per dose:
10
Details on study design:
1st application: Induction 0.5 % intracutaneous
2nd application: Induction 100 % occlusive epicutaneous
3rd application: Challenge 25 % occlusive epicutaneous
Challenge controls:
Eight animals were treated as controls and received induction and challenge treatments similar to the test pigs minus the test material.

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25% Galaxolide
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
very faint erythema (score 0.5)
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25% Galaxolide. No with. + reactions: 2.0. Total no. in groups: 10.0. Clinical observations: very faint erythema (score 0.5).
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25% Galaxolide
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
very faint erythema (score 0.5)
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25% Galaxolide. No with. + reactions: 3.0. Total no. in groups: 10.0. Clinical observations: very faint erythema (score 0.5).
Reading:
rechallenge
Group:
test group
Dose level:
25% Galaxolide
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. Group: test group. Dose level: 25% Galaxolide. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25% Galaxolide
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
very faint erythema (Score 0,5)
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25% Galaxolide. No with. + reactions: 3.0. Total no. in groups: 10.0. Clinical observations: very faint erythema (Score 0,5).
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25% Galaxolide
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
very faint erythema (score 0,5)
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25% Galaxolide. No with. + reactions: 1.0. Total no. in groups: 10.0. Clinical observations: very faint erythema (score 0,5).
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25% Galaxolide
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
very faint erythma (score 0,5)
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25% Galaxolide. No with. + reactions: 1.0. Total no. in groups: 10.0. Clinical observations: very faint erythma (score 0,5).
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25% Galaxolide
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25% Galaxolide. No with. + reactions: 0.0. Total no. in groups: 10.0.

Any other information on results incl. tables

At 24 hours, very faint erythema (score 0.5) was found in 2/10 animals at challenge 1, 3/10 animals at challenge 2, and 1/10 at challenge 3. At 48 hours, 3/10, 1/10 and 0/10 had very faint erythema. At challenge at 24 hours, only one animal showed very faint erythema to faint erythema.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
Except for one equivocal response in one animal, no evidence that the material was a sensitiser was seen. Classification: not sensitizing
Executive summary:

Galaxolide (65% HHCB in DEP) has been subjected to a non-GLP guinea pig maximization test. The used doses of Galaxolide were 0.5% in 0.01% dodecylbenzene sulphonate in 0.9% saline (DOBS/saline) for the intradermal injection, 100% for the induction patch, and 25% in 70% acetone/30% polyethylene glycol 400 (acetone/PEG400) for the challenge patch. These doses were selected based on preliminary irritation tests using 0.1, 0.25, 0.5, 1.0 and 2.0% Galaxolide concentrations for intradermal injections, however the selection criteria were not clear. The actual concentrations of HHCB are 0.325%, 65%, and 16.25%, respectively. Ten (six male, four female) Albino Dunkin/Hartley guinea pigs (weight 316-350g) were tested on a 2cm x 4cm area of skin in the dorsal shoulder area, clipped free of fur. Induction consisted of a 0.1 ml intradermal injection of 0.325% HHCB in DOBS/saline and 0.1 ml 50% Freund’s Complete Adjuvant in 0.9% saline. This was followed one week later by a 48 hr occluded patch (filter paper attached by adhesive tape to polythene backing) saturated with 65% HHCB). The patch was applied at the same 2 cm by 4cm area after freshly shaving the skin. Challenge applications were made 14 days later at a freshly shaved naïve site by saturation of an 8mm diameter filter paper patch with 16.25% HHCB in 70% acetone/30% PEG 400. Eight animals were treated as controls and received induction and challenge treatments similar to the test pigs minus the test material. Two repeat challenges at weekly intervals were conducted. At 24 hours, very faint erythema (score 0.5) was found in 2/10 animals at challenge 1, 3/10 animals at challenge 2, and 1/10 at challenge 3. At 48 hours, 3/10, 1/10 and 0/10 had very faint erythema. At challenge at 24 hours, only one animal showed very faint erythema to faint erythema. Except for one equivocal response in one animal, no evidence that the material was a sensitiser was seen (Basketter, 1996).