Registration Dossier

Administrative data

Description of key information

Key value for chemical safety assessment

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:
1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Meets generally accepted scientific standards, well documented and acceptable for assessment.
Qualifier:
equivalent or similar to
Guideline:
other: Magnusson B, Kligman AM (1970) Allergic contact dermatitis in the guinea pig: Identification of contact allergens. CC Thomas, Springfield, IL
Principles of method if other than guideline:
Guinea pig maximisation test; induction via intradermal and occluded patch; 4 challenge exposures at different concentrations on different days.
GLP compliance:
no
Remarks:
Not fully compliant with GLP because the test substance was not subjected to characterisation, homogeneity and stability studies.
Type of study:
guinea pig maximisation test
Species:
guinea pig
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: No data
- Age at study initiation: No data
- Weight at study initiation: No data
- Housing: No data
- Diet (e.g. ad libitum): No data
- Water (e.g. ad libitum): No data
- Acclimation period: No data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data
- Humidity (%): No data
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data
Route:
intradermal and epicutaneous
Vehicle:
other: Intradermal injections - 0.1 % dodecylbenzene sulphonate (DOBS) in 0.9 % physiological saline; occluded patch applications - none
Concentration / amount:
Intradermal injection induction - 2.0 %; occluded patch induction application - 100 %; occluded patch first challenge application - 100 %
Route:
epicutaneous, occlusive
Vehicle:
other: Intradermal injections - 0.1 % dodecylbenzene sulphonate (DOBS) in 0.9 % physiological saline; occluded patch applications - none
Concentration / amount:
Intradermal injection induction - 2.0 %; occluded patch induction application - 100 %; occluded patch first challenge application - 100 %
No. of animals per dose:
Test group - 10; treated control group - 4; untreated control group - 4
Details on study design:
PRELIMINARY INTRADERMAL INJECTION TEST
- No. of animals: 4 female
- Exposure period: 24 hr
- Observations: skin reactions examined, reactions measured in size.
- 2 % test substance in 0.01 % DOBS/saline was selected for intradermal injection induction as it was suitably irritant.

PRELIMINARY OCCLUDED PATCH TEST
- No. of animals: 8 male in 2 groups of 4.
- Exposure period: 24 hr
- Observation period: 48 hr, observations at 24 hr and 48 hr.
- Concentrations: Group 1 - 5.0 %, 10.0 %, 25.0 %. Group 2 - 50.0 %, 100.0 %
- 100 % test substance was selected for both the induction and challenge applications.

MAIN STUDY
- No. of animals: 10 (6 male, 4 female).
- Weight at study initiation: 284 g to 370 g.

A. INTRADERMAL INJECTION INDUCTION
- No. of exposures: 1
- Site: Hair clipped from 2 cm × 4 cm region of dorsal shoulder.
- Injections into the prepared site as follows:
-- 2 × 0.1 mL injections of 50 % Freund's Complete Adjuvant (FCA) in 0.9 % saline
-- 2 × 0.1 mL injections of 2 % test substance in 0.01 % DOBS/saline
-- 2 × 0.1 mL injections of 4 % test substance in 0.01 % DOBS/saline mixed 50:50 with FCA so that concentration of test substance was 2 %.

B. OCCLUDED PATCH INDUCTION
- Period: 1 week after Intradermal Injection Induction
- Site: same site clipped and shaved
- 2 cm × 4 cm filter paper patch, attached to a 4 cm × 6 cm piece of polythene, was saturated with neat test substance and patch applied to site.
- Exposure period: 48 hr

C. CHALLENGE EXPOSURES
- No. of challenge exposures: 4
- Days of challenges:
-- 1: day 0 (14 days after the occluded patch induction)
-- 2: day 7
-- 3: day 15
-- 4: day 22
- Concentrations:
-- 1: 100 % test substance
-- 2: 10.0 % and 1.0 % test substance in acetone/PEG 400
-- 3: 0.1 % and 0.001 % test substance in acetone/PEG 400
-- 4: 1.0 % and 0.1 % test substance in acetone/PEG 400
- Site: same site clipped and shaved
- 8 mm diameter filter paper patch in an 11 mm aluminium patch test cup was saturated with challenge material and patch applied to site.
- Exposure period: 24 hr
- Evaluations: 24 hrs and 48 hrs after challenge.
- Scoring system:
-- 0: No reaction
-- 0.5: Very feint erythema (usually non-confluent)
-- 1: Faint erythema (usually confluent)
-- 2: Moderate erythema
-- 3: Marked erythema (with or without oedema)
-- sp: Small spots of erythema
-- n: Necrosis
-- w: White centre or spots
Challenge controls:
- No. of animals: 16
-- 4 treated controls for 1st challenge
-- 4 treated controls for 2nd challenge.
-- 4 untreated controls for 3rd challenge.
-- 4 untreated controls for 4th challenge.
- Treated animals received four intradermal injections of 50 % Freunds Complete Adjuvant in 0.01 % DOBS/saline followed 7 days later by a 48 hour occluded patch of the test solvent over the injection sites.
- Each group of 4 animals was treated to 1st, 2nd, 3rd and 4th challenge exposures as per the test animals.
Positive control substance(s):
no
Reading:
other: 1st challenge (day 0)
Hours after challenge:
24
Group:
test group
Dose level:
100.0 %
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
other: Reading: other: 1st challenge (day 0). . Hours after challenge: 24.0. Group: test group. Dose level: 100.0 %. No with. + reactions: 8.0. Total no. in groups: 10.0.
Reading:
other: 1st challenge (day 0)
Hours after challenge:
48
Group:
test group
Dose level:
100.0 %
No. with + reactions:
9
Total no. in group:
10
Remarks on result:
other: Reading: other: 1st challenge (day 0). . Hours after challenge: 48.0. Group: test group. Dose level: 100.0 %. No with. + reactions: 9.0. Total no. in groups: 10.0.
Reading:
other: 2nd challenge (day 7)
Hours after challenge:
24
Group:
test group
Dose level:
10.0 %
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
other: Reading: other: 2nd challenge (day 7). . Hours after challenge: 24.0. Group: test group. Dose level: 10.0 %. No with. + reactions: 8.0. Total no. in groups: 10.0.
Reading:
other: 2nd challenge (day 7)
Hours after challenge:
48
Group:
test group
Dose level:
10.0 %
No. with + reactions:
7
Total no. in group:
10
Remarks on result:
other: Reading: other: 2nd challenge (day 7). . Hours after challenge: 48.0. Group: test group. Dose level: 10.0 %. No with. + reactions: 7.0. Total no. in groups: 10.0.
Reading:
other: 2nd challenge (day 7)
Hours after challenge:
24
Group:
test group
Dose level:
1.0 %
No. with + reactions:
9
Total no. in group:
10
Remarks on result:
other: Reading: other: 2nd challenge (day 7). . Hours after challenge: 24.0. Group: test group. Dose level: 1.0 %. No with. + reactions: 9.0. Total no. in groups: 10.0.
Reading:
other: 2nd challenge (day 7)
Hours after challenge:
48
Group:
test group
Dose level:
1.0 %
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
other: Reading: other: 2nd challenge (day 7). . Hours after challenge: 48.0. Group: test group. Dose level: 1.0 %. No with. + reactions: 8.0. Total no. in groups: 10.0.
Reading:
other: 3rd challenge (day 15)
Hours after challenge:
24
Group:
test group
Dose level:
0.1 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: other: 3rd challenge (day 15). . Hours after challenge: 24.0. Group: test group. Dose level: 0.1 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
other: 3rd challenge (day 15)
Hours after challenge:
48
Group:
test group
Dose level:
0.1 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: other: 3rd challenge (day 15). . Hours after challenge: 48.0. Group: test group. Dose level: 0.1 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
other: 3rd challenge (day 15)
Hours after challenge:
24
Group:
test group
Dose level:
0.001 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: other: 3rd challenge (day 15). . Hours after challenge: 24.0. Group: test group. Dose level: 0.001 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
other: 3rd challenge (day 15)
Hours after challenge:
48
Group:
test group
Dose level:
0.001 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: other: 3rd challenge (day 15). . Hours after challenge: 48.0. Group: test group. Dose level: 0.001 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
other: 4th challenge (day 22)
Hours after challenge:
24
Group:
test group
Dose level:
1.0 %
No. with + reactions:
7
Total no. in group:
10
Remarks on result:
other: Reading: other: 4th challenge (day 22). . Hours after challenge: 24.0. Group: test group. Dose level: 1.0 %. No with. + reactions: 7.0. Total no. in groups: 10.0.
Reading:
other: 4th challenge (day 22)
Hours after challenge:
48
Group:
test group
Dose level:
1.0 %
No. with + reactions:
6
Total no. in group:
10
Clinical observations:
1 additional questionable reaction
Remarks on result:
other: Reading: other: 4th challenge (day 22). . Hours after challenge: 48.0. Group: test group. Dose level: 1.0 %. No with. + reactions: 6.0. Total no. in groups: 10.0. Clinical observations: 1 additional questionable reaction.
Reading:
other: 4th challenge (day 22)
Hours after challenge:
24
Group:
test group
Dose level:
0.1 %
No. with + reactions:
4
Total no. in group:
10
Remarks on result:
other: Reading: other: 4th challenge (day 22). . Hours after challenge: 24.0. Group: test group. Dose level: 0.1 %. No with. + reactions: 4.0. Total no. in groups: 10.0.
Reading:
other: 4th challenge (day 22)
Hours after challenge:
48
Group:
test group
Dose level:
0.1 %
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
1 additional questionable reaction
Remarks on result:
other: Reading: other: 4th challenge (day 22). . Hours after challenge: 48.0. Group: test group. Dose level: 0.1 %. No with. + reactions: 3.0. Total no. in groups: 10.0. Clinical observations: 1 additional questionable reaction.

Strong skin sensitisation was seen with challenges of 100.0 %, 10.0 % and 1.0 % test material solution, a weak response was seen at 0.1 %, and no response was seen at 0.001 %.

There were no reactions observed in the treated control groups for challenges 1 and 2. One animal in the group for challenge 2 was killed and not scored as it suffered a broken leg. There were also no reactions observed in the untreated control group for challenges 3 and 4.

Interpretation of results:
sensitising
Remarks:
Migrated information Criteria used for interpretation of results: not specified
Conclusions:
The test material was shown to be strongly sensitising in a guinea pig maximisation test.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

In Unilever study No. D97/056, a non-guideline guinea pig maximisation test, the substance was found to be sensitising to skin. The results of any further testing are also likely to be positive and may therefore cause unnecessary pain and suffering of animals. Further testing therefore can be discounted on animal welfare grounds.


Migrated from Short description of key information:
Sensitising to skin (guinea pig maximisation test).

Justification for selection of skin sensitisation endpoint:
Key study.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Skin sensitisation

The substance shall be classified as a skin sensitiser category 1 in accordance with Regulation (EC) 1272/2008, Annex I, section 3.4.2.2., specifically section 3.4.2.2.2.1., criterion (c); "positive data from appropriate animal studies". Specifically, section 3.4.2.2.4.1 states that "When an adjuvant type guinea pig test method for skin sensitisation is used, a response of at least 30 % of the animals is considered as positive".

Respiratory sensitisation

According to Regulation (EC) 1272/2008, Annex I, section 3.4.2.1, "Substances shall be classified as respiratory sensitisers (Category 1) in accordance with the following criteria:(a) if there is evidence in humans that the substance can lead to specific respiratory hypersensitivity and/or (b) if there are positive results from an appropriate animal tests." In this case, due to the lack of positive human evidence, and the lack of appropriate animal tests, the substance should be classified as "data lacking" for respiratory sensitisation.