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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:
other: experimental study on similar substance
Adequacy of study:
key study
Study period:
1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
The room temperature varied between 21°C and 25°C during the study procedure.
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The test was performed according to a generally accepted method.
Species:
guinea pig
Strain:
Himalayan
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS- Number of animlas: 30 males distributed as: 20 main test, 10 control group 6 males for the pre-test distributed as: 2 for intradermal test and 4 for epidermal test - Source: BRL, Biological Research Laboratories Ltd. Wölferstrasse 4, 4414 Füllinsdorf/Switzerland- Weight at study initiation: 306-377 g test group / 295-430 g Positive control group - Housing: Individually in Makrol on type-3 cages with autoclaved standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz)- Diet : Diet: Pelleted standard Kliba 342, Batch no. 63/94 guinea pig breeding/ maintenance diet ("Kliba", Klingentalmühle AG, CH-4303 Kaiseraugst), ad libitum. Results of analyses for contaminants are included in this report.- Water: Community tap water from Fiillinsdorf, ad libitum. Once weekly additional supply of ascorbic acid (1 g/1) via the drinking water. Results of bacteriological, chemical and contaminant analyses are included in this report.- Acclimation period: One week for the control and test group under test conditions after health examination. No acclimatization for the animals of the pretest. Only animals without any visual signs of illness were used for the study.Regular cleaning and disinfection of cages was performed to prevent contamination of pathogens according to standard operating procedure.ENVIRONMENTAL CONDITIONS- Temperature (°C): 21° C - 25°C- Humidity (%): 52% -70%- Photoperiod: 12-hour light cycle- Music during the light period
Route:
intradermal and epicutaneous
Vehicle:
other: water for intradermal injections and vaselinum alba for epidermal application
Concentration / amount:
For injections: 5 %,of the tested substance in water solutionFor topic application: for induction peridod 50% of tested substance in vaseline alba, for the challenge 25 % of tested substance in vaseline alba
Route:
epicutaneous, occlusive
Vehicle:
other: water for intradermal injections and vaselinum alba for epidermal application
Concentration / amount:
For injections: 5 %,of the tested substance in water solutionFor topic application: for induction peridod 50% of tested substance in vaseline alba, for the challenge 25 % of tested substance in vaseline alba
No. of animals per dose:
Experimental group: 20 males Control group: 10 males
Details on study design:
A. INDUCTION: INTRADERMAL INJECTIONDay 1- treated group: 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 as follows: injection 1: a 1:1 mixture Freunds Complete Adjuvant (FCA)/physiological saline injection 2: 5 % of the test substance in bi-distilled water solution injection 3: 5 % of the test substance in water solution formulated in a 1:1 mixture FCA/waterDay 1- control group: Three paires of intradermal injections of 0,1 ml volume were applied to the shaved area. injection 1: a 1:1 mixture FCA/physiological saline injection 2: bi-distilled water for injection injection 3: 1:1 (w/w) mixture of bi-distilled water in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline. INDUCTION: TOPICAL APPLICATIONDay 8- treated group: Tested area was again shaved on day 7. On the identical site of intradermal injection was applied a filter paper (2x4cm) which was loaded with 50% of tested substance in white vaseline and held in contact by an occlusive dressing. The patch was covered with aluminum foil and firmly secured by an elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape. The dressings were left in place for approximately 48 hours. The epidermal application procedure described ensured intensive contact of the test article. Day 8- control group: Tested area was again shaved. White vaseline was applied to an identical area of intradermal injection on filter paper (2x4 cm) in occlusive dressing for 48 hours. The gauze pad was kept in contact with the skin by an adhesive hypoalle rgenic patch under an occlusive aluminum foil sheet. B. CHALLENGE: TOPICAL APPLICATIONDay 22- treated and control group: On the right side of the animals was applied a filter paper (2x2 cm) impregnated with 25% solution of the tested substance in water for injections, occlusive dressing covered a period of 24hrs. After 24 and 48 after the patch removal, the challenge area was cleaned with water and was carried out clinical trials, focusing on the intensity of erythema and edema.
Positive control substance(s):
yes
Remarks:
2-mercaptobenzothiazol; 4-aminobenzoic acid ethyl ester
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25% in vaselinum album
No. with + reactions:
8
Total no. in group:
20
Clinical observations:
Erythema
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25% in vaselinum album. No with. + reactions: 8.0. Total no. in groups: 20.0. Clinical observations: Erythema.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
25 % in vaseline album
No. with + reactions:
5
Total no. in group:
20
Clinical observations:
Erythema
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25 % in vaseline album. No with. + reactions: 5.0. Total no. in groups: 20.0. Clinical observations: Erythema.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
-
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Erythema
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Erythema.
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
-
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Erythema
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Erythema.
Reading:
2nd reading
Hours after challenge:
24
Group:
test group
Dose level:
25% in vaseline album
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Oedema
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25% in vaseline album. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: Oedema.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25% in vaseline album
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Oedema
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25% in vaseline album. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: Oedema.
Reading:
2nd reading
Hours after challenge:
24
Group:
negative control
Dose level:
-
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Oedema
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 24.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Oedema.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
-
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Oedema
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Oedema.
Reading:
1st reading
Group:
positive control
Remarks on result:
positive indication of skin sensitisation

Pathology - Necropsy: No necropsies were performed on the animals sacrificed at termination of the observation period.

The animals were sacrificed with an intraperitoneal injection of NARCOREN (Rhone Merieux GmbH, D-88471 Laupheim) at a dose of at least 5.1 ml/kg body weight (equivalent to 810 mg sodium pentobarbitone/kg body weight) and discarded.

Clinical signs: No symptoms of systemic toxicity were observed in the animals.

Body weight: The body weight of the animals was within the normal range of variability. Three animals of the test group and three of the

epidermal pretest lost weight during the acclimatization period. They had recovered during the treatment period.

Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Conclusions:
The tested substance is a skin sensitizer.
Executive summary:

In this study 40% of the animals were positive at the 24-hour reading and 25% at the 48-hour reading after treatment with a non-irritant test substance concentration of 25% in vaselinum album.

The response of at least 30% positive animals is considered positive and the substance needs to be classified as H317 following the Regulation (CE) 1272/2008.







Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

The response of at least 30% positive animals is considered positive "R43" following the "Commission Directive 93/21/EEC, April 27, 1993 adapting to technical progress for the 18th time Council Directive 67/548/EEC on the approximation of the laws, regulations and administrative provisions relating to the classification, packaging and labelling of dangerous substances".

Therefore the tested substance should be considered to be a sensitizer.


Justification for selection of skin sensitisation endpoint:
The study was performed on a tested substance with a higher content of active ingredient.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Acid Blue 113 should be considered to be a sensitizer based on Regulation CE 1272/2008.