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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Existing study well reported and performed under GLP.
Species:
guinea pig
Strain:
Himalayan
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: BRL Biological Research laboratory Ltd- number of animals: 30 males / 6 females- Age at study initiation: 5 weeks- Weight at study initiation: Control and test group: 281-333 g Pretest:286-307 g- Housing: Individually in Makrolon type-3 cages with standard softwood bedding - Diet : Pelleted standard Kliba 342, Batch 63/91 guinea pig breeding/ maintenance diet, ad libitum.- Water: tap water , ad libitum. Once weekly additional supply of ascorbic acid (1 g/l) via the drinking water. - Acclimation period: One week under test conditions after veterinary examination.ENVIRONMENTAL CONDITIONS- Temperature (°C):22 ± 3°C- Humidity (%):40-70 % - Photoperiod : 12 hours artificial fluorescent light/12 hours dark, music/light period.
Route:
intradermal
Vehicle:
other: for induction: physiological saline; for epidermal: vaselina album
Concentration / amount:
5% for intradermal injenction5, 10, 15 and 25% for epidermal application The highest non-irritating test article concentration used for challenge application was 15%.
Route:
epicutaneous, occlusive
Vehicle:
other: for induction: physiological saline; for epidermal: vaselina album
Concentration / amount:
5% for intradermal injenction5, 10, 15 and 25% for epidermal application The highest non-irritating test article concentration used for challenge application was 15%.
No. of animals per dose:
Main study induction: Control 10 females / Test Group: 20 femalesMain study challenge: Control 10 females / Test Group: 20 females
Details on study design:
MAIN STUDYA. INDUCTION EXPOSURE: injection- Test groups: 20 females- Control group: 10 females- Site: An area of dorsal skin from the scapular region (approximately 6 x 8 cm) was clipped free of hair. Three pairs of intradermal injections (0.1 ml/site) were made at the border of a 4 x 6 cm area in the clipped region- Concentrations: 5% with physiological saline, 5% by emulsion in a 50:50 mixture of Freund's colplete adjuvant and physiological salineINDUCTION EXPOSURE: epidermal application, one week after the injections- Exposure period: 48 h- Test groups: 20 females- Control group: 10 females- Site: (6 x 8 cm)- Type of coverage: filter paper was saturated with the test article and placed over the injection sites of the test animais. The patch was covered by aluminum foil and firmly secured by an elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape.- Concentrations: 25 % in vaseline albumB. CHALLENGE EXPOSURE: The test and control guinea-pigs were challenged two weeks after the epidermal induction application.- Exposure period: approximately 24 hours later- Test groups: 20 females- Control group: 10 females- Site:hair was clipped and shaved from a 5 x 5 cm area on the left and right flank of each guinea-pig.- Type of coverage: Two patches (2 x 2 cm) of filter paper- Concentrations: 15% in vaseline album - Evaluation : 24, 48 hrsOTHER OBSERVATION:Mortality/Viability: dailyBody Weights:At acclimatization start, start of application and end of test.Symptoms (local systemic): dailyNecropsy: noAll animals were killed at the end of the test period with an intraperitoneal injection of pentobarbital (> 800 mg/kg) and discarded.
Positive control substance(s):
yes
Remarks:
Formaldheyde solution
Positive control results:
8 guinea pigs in a group of 10 animals showed positive response to the formaldehyde solution.
Reading:
1st reading
Hours after challenge:
28
Group:
test chemical
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No systemic symptoms were observed.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 28.0. Group: test group. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No systemic symptoms were observed..
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No systemic symptoms were obseved.
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No systemic symptoms were obseved. .
Reading:
1st reading
Group:
negative control
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
positive control
Remarks on result:
positive indication of skin sensitisation
Interpretation of results:
GHS criteria not met
Conclusions:
According to EEC (European Economie Community) and CLP classification criteria described in guidelines 83/467, September 16, 1983, the tested substance Direct Red 81 is not a sensitizer.
Executive summary:

The purpose of this skin sensitization study was to assess the allergenic potential of Direct Red 81 when administered to the skin of albino guinea pigs following OECD 406, Maximization-Test of B. Magnusson and A.M. Kligman (1969).

Ten animals (10 females) were treated with the vehicle alone (vaseline album) and 20 animals (20 females) were treated with the test article.

Due to the unequivocal findings observed after the first challenge, no second challenge was performed.

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

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

SKIN SENSITIZATION

Category 1

Substances shall be classified as skin sensitizers in category 1 where data are not sufficient for sub-categorisation in accordance with the following criteria:

(a) if there is evidence in humans that the substance can lead to sensitisation by skin contact in a substantial number of persons; or

(b) if there are positive results from an appropriate animal test.

Specific criteria of animal test:

when an adjuvant type test method for skin sensitisation is used, a response of at least 30 % of the animals is considered as positive.

For a non-adjuvant Guinea pig test method a response of at least 15 % of the animals is considered positive.

Furthermore, stimulation index of three or more is considered a positive response in the local lymph node assay.

Sub-category 1A

Substances showing a high frequency of occurrence in humans and/or a high potency in animals can be presumed to have the potential to produce significant sensitisation in humans. Severity of reaction may also be considered.

Specific criteria:

Local lymph node assay-EC3 value ≤ 2 %

Guinea pig maximisation test-≥ 30 % responding at ≤ 0,1 % intradermal induction dose or ≥ 60 % responding at > 0,1 % to ≤ 1 % intradermal induction dose

Buehler assay - ≥ 15 % responding at ≤ 0,2 % topical induction dose or ≥ 60 % responding at > 0,2 % to ≤ 20 % topical induction dose

Sub-category 1B

Substances showing a low to moderate frequency of occurrence in humans and/or a low to moderate potency in animals can be presumed to have the potential to produce sensitisation in humans. Severity of reaction may also be considered.

Local lymph node assay - EC3 value > 2 %

Guinea pig maximisation test- ≥ 30 % to < 60 % responding at > 0,1 % to ≤ 1 % intradermal induction dose or ≥ 30 % responding at > 1 % intradermal induction dose.

Buehler assay - ≥ 15 % to < 60 % responding at > 0,2 % to ≤ 20 % topical induction dose or ≥ 15 % responding at > 20 % topical induction dose.

 

Based on the results obtained in the in vivo test and according to the paragraph 3.4. of the CLP Regulation n. 1272/2008, the test substance is NOTclassified.