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

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

From February 9 to 12,1993.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guidelines and performed according to GLP. 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:

1993

Report date:

1993

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1981

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

224-073-5

EC Name:

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

Cas Number:

4193-55-9

IUPAC Name:

disodium 5-({4-anilino-6-[bis(2-hydroxyethyl)amino]-1,3,5-triazin-2-yl}amino)-2-{2-[4-({4-[bis(2-hydroxyethyl)amino]-6-(cyclohexa-1,3-dien-1-ylamino)-1,3,5-triazin-2-yl}amino)-2-sulfonatophenyl]vinyl}benzenesulfonate

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd., Moston, Sandbach, Cheshire, U.K.
- Age at study initiation: twelve to sixteen weeks old.
- Weight at study initiation: 2.53 - 2.79 kg.
- Housing: individually housed in suspended metal cages.
- Diet: free access to food (RABMA Rabbit Diet, Special Diet Services Ltd., witham, Essex, U.K.).
- Water: free access to mains drinking water.
- Acclimation period: a minimum acclimatisation period of five days.

ENVIRONMENTAL CONDITIONS
- Temperature: 18 - 21°C
- Humidity: 39 - 45 %.
- Air changes: approximately 15 changes per hour.
- Photoperiod: the lighting was controlled by a time switch to give continuous 12 hours light and 12 hours darkness.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.5 ml of the test material was introduced under a 2.5 cm x 2.5 cm gauze patch and placed in position on the shorn skin.

Number of animals:

Three

Details on study design:

TEST SITE
- Area of exposure: on the day before the test each of a group of three rabbits was clipped free of fur from the dorsal flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study. On the day of the test a suitable test site was selected on the back of each rabbit.
- Type of wrap if used: the patch was secured in position with a strip of surgical adhesive tape (BLENDERM: approximate size 2.5 cm x 4.0 cm). To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset (TUBIGRIP) and the animals were returned to their cages for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing: the corset and patches were removed from each animal and any residual test material removed by gentle swabbing with cotton wool soaked in distilled water.
- Time after start of exposure: four hours after application.

SCORING SYSTEM
Approximately one hour following the removal of the patches, and 24,48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the following scale from Draize J.H. (1959) Association of Food and Drug Officials of the United States, Austin, Texas, "The Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics":
Erythema and Eschar Formation
No erythema: 0
Very slight erythema (barely perceptible): 1
Well-defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth): 4

Oedema Formation
No oedema: O
Very slight oedema (barely perceptible): 1
Slight oedema (edges of area well-defined by definite raising): 2
Moderate oedema (raised approximately 1 millimetre): 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure): 4

Any other skin reactions, if present, were also recorded.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

other: mean 24, 48 and 72 hours

Score:

< 2.3

Reversibility:

fully reversible

Results 2

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

other: mean 24, 48 and 72 hours

Score:

< 2.3

Reversibility:

fully reversible

Irritant / corrosive response data:

The test material produced a primary irritation index of 0.3. According to the Draize classification scheme: mild irritating.
According to the CLP regulation: not irritating.

Any other information on results incl. tables

Skin reaction	Reading (hrs)	Individual scores
		147 F	156 F	161 F
Erythema/eschar formation	24	0	1	1
	48	0	0	0
	72	0	0	0
Total		0	1	1
Mean score		0	0.3	0.3
Oedema formation	24	0	0	0
	48	0	0	0
	72	0	0	0
Total		0	0	0
Mean score		0	0	0

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

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

Conclusions:

Not irritating.

Executive summary:

Method

A study was performed to assess the irritancy potential of the test material to the skin of the New Zealand white rabbit. The method used followed that described in the OECD Guidelines No. 404 "Acute Dermal Irritation/Corrosion" referenced as Method B4 in Commission Directive 84/449/EEC.

Results

A single 4 -hours, semi-occluded application of the test material to the intact skin of three rabbits produced well-defined erythema and very slight oedema. All treated skin sites appeared normal at the 48-hour observation.

The test material produced a primary irritation index of 0.3 and was classified as a mild irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

Conclusion

According to the CLP regulation (EC 1272/2008), test substance can be classified as non irritating, because mean value scored is less than 2.3 for erythema/eschar and for oedema in at least 2 of 3 tested animals from gradings at 24, 48 and 72 hours after patch removal.