Reaction mass of 1H-Benzotriazole-1-methanamine, N,N-bis(2-ethylhexyl)-6-methyl- and 2H-Benzotriazole-2-methanamine, N,N-bis(2-ethylhexyl)-5-methyl- and N,N-bis(2-ethylhexyl)-4-methyl-1H-benzotriazole-1-methylamine and 2H-Benzotriazole-2-methanamine, N,N-bis(2-ethylhexyl)-4-methyl- and N,N-bis(2-ethylhexyl)-5-methyl-1H-benzotriazole-1-methylamine

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 7, 1990 - February 7, 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The study was conducted in 1991, when the GPMT was the internationally accepted and recommended study type in order to assess sensitizing potential of a substance.

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 1H-Benzotriazole-1-methanamine, N,N-bis(2-ethylhexyl)-6-methyl- and 2H-Benzotriazole-2-methanamine, N,N-bis(2-ethylhexyl)-5-methyl- and N,N-bis(2-ethylhexyl)-4-methyl-1H-benzotriazole-1-methylamine and 2H-Benzotriazole-2-methanamine, N,N-bis(2-ethylhexyl)-4-methyl- and N,N-bis(2-ethylhexyl)-5-methyl-1H-benzotriazole-1-methylamine

EC Number:

939-700-4

Molecular formula:

Unspecified

IUPAC Name:

Reaction mass of 1H-Benzotriazole-1-methanamine, N,N-bis(2-ethylhexyl)-6-methyl- and 2H-Benzotriazole-2-methanamine, N,N-bis(2-ethylhexyl)-5-methyl- and N,N-bis(2-ethylhexyl)-4-methyl-1H-benzotriazole-1-methylamine and 2H-Benzotriazole-2-methanamine, N,N-bis(2-ethylhexyl)-4-methyl- and N,N-bis(2-ethylhexyl)-5-methyl-1H-benzotriazole-1-methylamine

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Pirbright White strain (Tif:DHP)

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: from Ciba Geigy Limited, Animal Production, 4332 Stein, Switzerland
- Age at study initiation: approximately 10 weeks
- Weight at study initiation: between 338 and 437 g
- Housing: housed individually in Macrolon cages (Type 3)
- Diet: standard guinea pig pellets (NAFAG no. 845, Gossau SG); ad libitum
- Water: ad libitum
- Acclimation period: 8 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 30 - 70%
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

other: sesame oil

Concentration / amount:

3 %

Day(s)/duration:

day 0

Induction 2

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Concentration / amount:

10 %

Day(s)/duration:

week 2 / 48 h

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Concentration / amount:

3 %

Day(s)/duration:

week 5 / 24 h

No. of animals per dose:

20 (10 males and 10 females)

Details on study design:

INDUCTION:
- 2-stage operation
- First induction: 3 pairs of intradermal injections (0.1 mL per injection) made simultaneously into the shaved neck of the guinea pigs as follows:
adjuvant/saline mixture 1:1 (v/v)
test article in sesame oil (3%) (w/v)
test article in the adjuvant saline mixture (w/v)
- Second induction: in the second week, the test article was incorporated in vaseline (10%) (w/w) and applied on a filterpaper patch to the neck of the animals (patch 2 x 4 cm, approximately 0.4 g paste per patch
Occluded administration for 48 hours.
A control group (20 animals: 10 males and 10 females) was treated with adjuvant and the vehicle during the induction period.

REST PERIOD:
no treatment performed during weeks 3 and 4

CHALLENGE:
Animals were tested on the flank with the test article in vaseline (3%) (w/w) or the vehicle alone (patch 2 x 2 cm, approximately 0.2 g paste per patch)
Occluded administration for 24 hours.
During the challenge period, the control group of the induction was treated with the vehicle as well as with the test article to check the maximum subirritant concentration of the test article in adjuvant treated animals.

Challenge controls:

A control group (20 animals: 10 males and 10 females) was treated with adjuvant and the vehicle during the induction period.
During the challenge period, the group was treated with the vehicle as well as with the test article to check the maximum subirritant concentration of the test article in adjuvant treated animals.

Positive control substance(s):

yes

Remarks:

sensitivity checked every 6 months: potassium dichromate

Results and discussion

Positive control results:

Incidence of positive animals per group after epidermal challenges 24 hours after removing of bandages:
7/10 for erythema
5/10 for edema

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

3%

No. with + reactions:

5

Total no. in group:

20

Clinical observations:

(effects only in females)

Results 2

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

3%

No. with + reactions:

5

Total no. in group:

20

Clinical observations:

(effects only in females)

Results 3

Key result

Reading:

rechallenge

Hours after challenge:

24

Group:

test chemical

Dose level:

3%

No. with + reactions:

8

Total no. in group:

20

Clinical observations:

(3 males, 5 females)

Results 4

Key result

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

3%

No. with + reactions:

7

Total no. in group:

20

Clinical observations:

(2 males, 5 females)

Results 5

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No toxic symptoms

Results 6

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No toxic symptoms

Results 7

Group:

positive control

Remarks on result:

not measured/tested

Any other information on results incl. tables

Results of test group:

Males		Animal number
Challenge		1	2	3	4	5	6	7	8	9	10
24 h	Erythema	0	0	0	0	0	0	0	0	0	0
	Edema	0	0	0	0	0	0	0	0	0	0
48 h	Erythema	0	0	0	0	0	0	0	0	0	0
	Edema	0	0	0	0	0	0	0	0	0	0
Re-Challenge
24 h	Erythema	1	2	0	1	0	0	0	0	0	0
	Edema	0	1	0	0	0	0	0	0	0	0
48 h	Erythema	2	2	0	0	0	0	0	0	0	0
	Edema	1	1	0	0	0	0	0	0	0	0

 

Females		Animal number
Challenge		1	2	3	4	5	6	7	8	9	10
24 h	Erythema	0	1	1	0	1	1	1	0	0	0
	Edema	0	1	2	0	1	1	1	0	0	0
48 h	Erythema	0	1	1	0	1	1	1	0	0	0
	Edema	0	0	2	0	1	1	1	0	0	0
Re-Challenge
24 h	Erythema	0	2	2	1	0	1	2	0	0	0
	Edema	0	1	1	0	0	0	1	0	0	0
48 h	Erythema	0	2s	2s	1	0	2	2s	0	0	0
	Edema	0	1	1	0	0	1	2	0	0	0

s = scaling

In both control groups (vehicle control and test article treated control), there were 0/20 animals with positive responses at 24 and 48 hours after challenge.

Under the experimental conditions employed, 25% of the animals of the test group showed skin reactions 24 and 48 hours after removing the dressings.

Because the reactions were not very strong and the number of positive animals was at the threshold of significance, a second challenge was performed after a further rest period. The control group was not challenged again.

After the second challenge application, the number of positive animals was increased to 40% and the reactions intensified.

Applicant's summary and conclusion

Interpretation of results:

Category 1B (indication of skin sensitising potential) based on GHS criteria

Conclusions:

25 to 40% of the animals were sensitised by the test substance under the experimental conditions employed.