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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1977
Report date:
1977

Materials and methods

Principles of method if other than guideline:
Optimization test*, an intracutaneous sensitization procedure similar to the method recommended in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959), the US Association of Food and Drug Officials (AFDO).

*Maurer, Th., Thomann, P., Weirich, E.G. and Hess, R., (1975). The Optimization test in the guinea pig. A method for the predictive evaluation of the contact allergenicity of Chemicals. Agents and Actions Vol. 5 (2), 174-179, 1975
GLP compliance:
no
Type of study:
Maurer optimisation test
Justification for non-LLNA method:
Old test

Test material

Constituent 1
Reference substance name:
Fluorescent Brightener 363
IUPAC Name:
Fluorescent Brightener 363

In vivo test system

Test animals

Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Weight at study initiation: between 350 to 400 grams.
- Housing: animals were housed individually in Macrolon cages, type 3.
- Diet: standard guinea pig pellets - NAFAG No. 830, Gossau SG - and fresh carrots, ad libitum.
- Water: ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 1 °C
- Relative humidity: 55 ± 5 %
- Photoperiod: 14 hours light cycle day.

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal
Vehicle:
physiological saline
Concentration / amount:
0.1 %
Challengeopen allclose all
Route:
intradermal
Vehicle:
physiological saline
Concentration / amount:
0.1 %
Route:
epicutaneous, occlusive
Vehicle:
other: vas. alb.
Concentration / amount:
1 %
Day(s)/duration:
24 hours
No. of animals per dose:
10 males and 10 females
Details on study design:
During the induction period the animals received one injection every second day (except weekends) to a total of 10 intracutaneous injections of a freshly prepared 0.1 % solution of test item in physiological saline.
One control group was treated with the vehicle alone ("negative control").
On the first day, injections of 0.1 ml were administered into the shaven skin of the right flank and the back, while on the following days a single intracutaneous injection was given into the back.

During the second and third week of the induction period the test compounds were incorporated in a mixture, of the normal vehicle with complete Bacto Adjuvant (vehicle : adjuvant = 1 : 1).

Fourteen days after the last sensitizing injection, a challenge injection of 0.1 ml of a freshly prepared 0. 1 % solution of test item was administered into the skin of the left flank.

Twenty-four hours after each injection during the first week of the induction period and 24 hours after the challenge injection the reactions were recorded.
Before examination, the reaction sites were depilated chemically (Veet, 5 minutes). The two largest perpendicular diameters (in mm) and the increase in the skin- fold thickness (in mm) were measured, and by multiplication of these values a "reaction volume" was obtained (in µl) for each reading from each animal. The mean volume plus one standard deviation of the induction reactions observed in the individual animal in the first week was taken as representing the skin irritation "threshold" for each animal. Any challenge reaction greater than this threshold value in the induction period was graded as an allergic reaction and the animal termed "positive". The number of "positive" animals in the test group was compared with the number of animals in the control group (treated with the vehicle alone) that showed a non-specific reaction of at least the same magnitude ("negative control").

Ten days after the intracutaneous challenge injection, a subirritant dose of the test compound was applied epicutaneously under occlusive dressings which were left in place for 24 hours.
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
0.1% - intradermal challenge
No. with + reactions:
2
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
1 % - epicutaneous challenge
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Dose level:
both intradermal and epicutaneous challenge
No. with + reactions:
2
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Group:
positive control
Remarks on result:
not measured/tested

Any other information on results incl. tables

No differences between the test group and the vehicle-treated controls were seen, after either intradermal or epidermal challenge application of test item.

Applicant's summary and conclusion

Interpretation of results:
other: not classified, according to the CLP Regulation (EC) No 1272/2008
Conclusions:
Not skin sensitising
Executive summary:

The skin sensitisation potential of test item was investigated in an optimation test proposed by Maurer et al (1973), an intracutaneous sensitization procedure similar to the method recommended in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959), the US Association of Food and Drug Officials (AFDO). The test was performed on groups of 10 male and 10 female guinea pigs. During the induction period the animals received one injection every second day (except weekends) to a total of 10 intracutaneous injections of a freshly prepared 0.1 % solution of test item in physiological saline. One control group was treated with the vehicle alone.

On the first day, injections of 0.1 ml were administered into the shaven skin of the right flank and the back, while on the following days a single intracutaneous injection was given into the back. During the second and third week of the induction period the test compounds were incorporated in a mixture, of the normal vehicle with complete Bacto Adjuvant (vehicle : adjuvant = 1 : 1). Fourteen days after the last sensitizing injection, a challenge injection of 0.1 ml of a freshly prepared 0.1 % solution of test item was administered into the skin of the left flank. Twenty-four hours after each injection during the first week of the induction period and 24 hours after the challenge injection the reactions were recorded. Before examination, the reaction sites were depilated chemically. The two largest perpendicular diameters (in mm) and the increase in the skin- fold thickness (in mm) were measured, and by multiplication of these values a "reaction volume" was obtained (in µl) for each reading from each animal. The mean volume plus one standard deviation of the induction reactions observed in the individual animal in the first week was taken as representing the skin irritation "threshold" for each animal. Any challenge reaction greater than this threshold value in the induction period was graded as an allergic reaction and the animal termed "positive". The number of "positive" animals in the test group was compared with the number of animals in the control group (treated with the vehicle alone) that showed a non-specific reaction of at least the same magnitude ("negative control"). Ten days after the intracutaneous challenge injection, a subirritant dose of the test compound was applied epicutaneously under occlusive dressings which were left in place for 24 hours. Two out of 20 animals were found to have a positive reaction after the intracutaneous challenge injection, while no positive reactions were recorded after the epicutaneous challenge application. Hence, based on the above information, it was concluded that substance was devoid of skin-sensitizing (contact allergenic) potential in albino guinea-pigs.

Conclusion

A response lower than 30 % was recorded, thus the substance does not meet the criteria to be classified as skin sensitizer, according to the CLP Regulation (EC) No 1272/2008.