Disodium 4,4'-bis[[4-anilino-6-[(2-hydroxyethyl)amino]-1,3,5-triazin-2-yl]amino]stilbene-2,2'-disulphonate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Study period:

From April 23 to June 15, 1979.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Internationally accepted guideline, well documented and scientifically acceptable. Justification for Read Across is detailed in the endpoint summary and in the Category Justification Report attached to the section 13.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Principles of method if other than guideline:

The optimization test was used, an intracutaneous sensitization procedure exceeding the sensitivity of 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).

GLP compliance:

no

Remarks:

Pre GLP

Type of study:

Maurer optimisation test

Test material

Test material information

Constituent 1

Reference substance name:

Tetrasodium 4,4'-bis[[4-[(2-hydroxyethyl)amino]-6-(m-sulphonatoanilino)-1,3,5-triazin-2-yl]amino]stilbene-2,2'-disulphonate

EC Number:

240-400-4

EC Name:

Tetrasodium 4,4'-bis[[4-[(2-hydroxyethyl)amino]-6-(m-sulphonatoanilino)-1,3,5-triazin-2-yl]amino]stilbene-2,2'-disulphonate

Cas Number:

16324-27-9

IUPAC Name:

tetrasodium 2,2'-ethene-1,2-diylbis[5-({4-[(2-hydroxyethyl)amino]-6-[(3-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)benzenesulfonate]

In vivo test system

Test animals

Species:

guinea pig

Strain:

Pirbright-Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: test laboratory.
- Age at study initiation: ca 10 weeks old.
- Weight at study initiation: between 280 to 390 grams.
- Housing: housed individually in Macrolon cages type 3.
- Diet: ad libitum standard guinea pig pellets - NAFAG, No. 830, Gossau SG.
- Water: ad libitum.
- Acclimation period: the animals were acclimatized for 10 days.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 1 °C
- Humidity: 55 ± 5 %
- Photoperiod: 14 hours light cycle day.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

other: intradermal: physiological saline; epidermal: vaseline PhH VI.

Induction 2

Concentration / amount:

0.1% for intradermal application
30% for epidermal application

Challenge

Challenge 1

Route:

intradermal and epicutaneous

Vehicle:

other: intradermal: physiological saline; epidermal: vaseline PhH VI.

Challenge 2

Concentration / amount:

0.1% for intradermal application
30% for epidermal application

No. of animals per dose:

1 male and 10 female guinea pigs

Details on study design:

A. INDUCTION EXPOSURE
- No. of exposures: one injection every second day (except weekends) to a total of 10 intracutaneous injections of a freshly prepared solution.
- Control group: one control group was treated with the vehicle alone ("negative control").
- Site: 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 material was incorporated in a mixture of the normal vehicle with complete Bacto Adjuvant.

B. CHALLENGE EXPOSURE
Intradermal
- First challenge injection: fourteen days after the last sensitizing injection.
- Site: administered into the skin of the left flank.
- Observetion: 24 hours after each injection during the first week of the induction period and 24 hours after the challenge injection the reactions were recorded.
- Evaluation/scoring system: 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 "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").

Epicutaneous
- Application: 10 days after the intracutaneous challenge injection
- Dose: subirritant dose of the test compound: 30 % in vaseline
- Route: test substance was applied epicutaneously under occlusive dressings.
- Duration/exposure: 24 hours.
- Scoring system: the reactions were evaluated 24 h after removing of the bandages according the Draize scoring scale.

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

other: Incidence of positive animals per group after intradermal challenge injection.

Group:

test chemical

Dose level:

0.1 %

No. with + reactions:

16

Total no. in group:

20

Remarks on result:

other: Reading: other: Incidence of positive animals per group after intradermal challenge injection.. Group: test group. Dose level: 0.1 % . No with. + reactions: 16.0. Total no. in groups: 20.0.

Results 2

Reading:

other: Incidence of positive animals per group after occlusive epicutaneous application.

Group:

test chemical

Dose level:

30 %

No. with + reactions:

2

Total no. in group:

20

Remarks on result:

other: Reading: other: Incidence of positive animals per group after occlusive epicutaneous application.. Group: test group. Dose level: 30 %. No with. + reactions: 2.0. Total no. in groups: 20.0.

Any other information on results incl. tables

Intradermal injection of the vehicle alone failed to induce sensitization.

Incidence of positive animals per group after intradermal challenge injection.

	N. of positive animals/N. treated animals	P
Vehicle control	0/20
Test substance	16/20	< 0.01

Incidence of positive animals per group after occlusive epicutaneous application.

	N. of positive animals/N. treated animals	P
Vehicle control	0/20
Test substance	2/20	> 0.01

Challenge reactions after occlusive epicutaneous administration of the test material.

Erythema score (Draize Score) 24 hours after removal of the dressing.

Animal N. males	681	682	683	684	685	686	687	688	689	690
Score	0	0	0	1	0	0	2	0	0	0
Animal N. females	691	692	693	694	695	696	697	698	699	700
Score	0	0	0	0	0	0	0	0	0	0

Reaction volumes (µl) after intradermal injection of test substance.

Animal identification number	Induction						(mean + s)	after skin sensitization	+ = positive reactor
	Application N.				Mean	Standard deviation
	1	2	3	4
681 M	0	0	0	0	0	0	0	0	-
682 M	0	0	0	0	0	0	0	32	+
683 M	0	0	0	0	0	0	0	9	+
684 M	0	0	0	0	0	0	0	224	+
685 M	0	0	0	0	0	0	0	65	+
686 M	0	0	9	0	2.3	4.5	6.8	64	+
687 M	0	0	0	0	0	0	0	62	+
688 M	0	0	0	0	0	0	0	6	+
689 M	0	0	0	0	0	0	0	18	+
690 M	0	0	0	0	0	0	0	7	+
691 F	0	0	0	0	0	0	0	72	+
692 F	0	0	0	0	0	0	0	0	-
693 F	0	0	0	0	0	0	0	0	-
694 F	0	0	0	0	0	0	0	79	+
695 F	0	0	0	0	0	0	0	45	+
696 F	0	0	0	0	0	0	0	112	+
697 F	0	0	0	0	0	0	0	315	+
698 F	0	0	0	0	0	0	0	13	+
699 F	5	0	0	0	1.3	2.5	3.8	170	+
700 F	0	0	0	0	0	0	0	0	-
Goup mean	0.3	0	0.5	0				64.7	16/20

Reaction volumes (µl) after intradermal injection of physiological saline.

Animal identification number	Induction						(mean + s)	after skin sensitization	+ = positive reactor
	Application N.				Mean	Standard deviation
	1	2	3	4
641 M	0	0	0	0	0	0	0	0	-
642 M	0	0	0	0	0	0	0	0	-
643 M	0	0	0	0	0	0	0	0	-
644 M	0	0	0	0	0	0	0	0	-
645 M	0	0	0	0	0	0	0	0	-
646 M	0	0	0	0	0	0	0	0	-
647 M	0	0	0	0	0	0	0	0	-
648 M	0	0	0	0	0	0	0	0	-
649 M	0	0	0	0	0	0	0	0	-
650 M	0	0	0	0	0	0	0	0	-
651 F	0	0	0	0	0	0	0	0	-
652 F	0	0	0	0	0	0	0	0	-
653 F	0	0	0	0	0	0	0	0	-
654 F	0	0	0	0	0	0	0	0	-
655 F	0	0	0	0	0	0	0	0	-
656 F	0	0	0	0	0	0	0	0	-
657 F	0	0	0	0	0	0	0	0	-
658 F	0	0	0	0	0	0	0	0	-
659 F	0	0	0	0	0	0	0	0	-
660 F	0	0	0	0	0	0	0	0	-
Goup mean	0	0	0	0				0	0/20

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information according to the CLP Regulation Criteria used for interpretation of results: EU

Conclusions:

Intradermal injection of the vehicle alone failed to induce sensitization.

Executive summary:

Method

The optimization test was used, an intracutaneous sensitization procedure exceeding the sensitivity of 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).

Results

Intradermal injection of the vehicle alone failed to induce sensitization.