Disodium 3-[[4-amino-9,10-dihydro-9,10-dioxo-3-[sulphonato-4-(1,1,3,3-tetramethylbutyl)phenoxy]-1-anthryl]amino]-2,4,6-trimethylbenzenesulphonate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November from 20 to 23, 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

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:

Acid Violet 048

IUPAC Name:

Acid Violet 048

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: approximately twelve to sixteen weeks old.
- Weight at study initiation: 2.19 - 2.26 kg
- Housing: animals were individually housed in suspended metal cages.
- Diet: Spillers Rabbit Diet (Dalgety Agriculture Ltd., Almondsbury, Bristol) ad libitum.
- Water: mains drinking water, ad libitum.
- Acclimation period: a minimum acclimatisation period of five days.

ENVIRONMENTAL CONDITIONS
- Temperature: 17 - 21 °C
- Humidity: 46 - 52 %
- Air changes: approximately 15 changes per hour.
- Photoperiod: lighting was controlled by a time switch to give 12 hours light and 12 hours darkness.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

The test material was moistened with distilled water immediately before application: 0.5 g of the test material, moistened with 0.5 ml of distilled water.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

1 male and 2 females

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. Test item was introduced under a 2.5 cm x 2.5 cm gauze patch and placed in position on the shorn skin.
- 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: any residual test material was removed by gentle swabbing with cotton wool soaked in distilled water followed by diethyl ether.
- Time after start of exposure: four hours after application the corset and patches were removed.

OBSERVATION TIME POINTS
Approximately one hour following the removal of the patches, and 24, 48 and 72 hours later, the test sites were examined.

SCORING SYSTEM
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 eryrhema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

Oedema formation:
No oedema 0
Very slight oedema (barely perceptible) 1
Well-defined oedema (area well defined by definite raising) 2
Moderate oedema (raised approximately 1 millimeter) 3
Severe oedema (raised more than 1 millimeter and extending beyond the area of exposure) 4

The scores for erythema and oedema at the 24 and 72-hour readings were totalled for the three test rabbits (12 values) and this total was divided by 6 to give the primary irritation index of the test material. The test material was classified according to the following scheme:
Non-irritant, primary irritation index 0
Mild irritant, primary irritation index >0 - 2
Moderate irritant > 2 - 5
Severe irritant > 5 - 8

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Reversibility:

fully reversible within: 72 hrs

Results 2

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Reversibility:

fully reversible within: 48 hrs

Irritant / corrosive response data:

Faint purple-coloured staining caused by the test material was commonly noted during the study. This did not prevent accurate evaluation of skin responses.
Very slight erythema was noted at two treated skin sites one hour after patch removal, at all treated skin sites at the 24-hour observation and at two treated skin sites at the 48-hour observation.
Very slight oedema was confined to two treated skin sites at the 24-hour observation.
No skin irritation was noted 72 hours after treatment.

Any other information on results incl. tables

Individual scores

Animal	Skin Reaction	After				Mean 24/48/72 hrs
		1 hr	24 hrs	48 hrs	72 hrs
171 F	Erythema	0 STA	1 STA	1 STA	0 STA	0.67
172 M	Erythema	1 STA	1 STA	0 STA	0 STA	0.33
175 F	Erythema	1 STA	1 STA	1 STA	0 STA	0.67
171 F	Oedema	0	1	0	0	0.33
172 M	Oedema	0	0	0	0	0.00
175 F	Oedema	0	1	0	0	0.33

Sum of 24 and 72 hours reading: 5

Primary irritation index: 0.8, mild irritant

Applicant's summary and conclusion

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Mild irritant

Executive summary:

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 for Testing of Chemicals (1981) No. 404 'Acute Dermal Irritation/Comosion' referenced as Method B4 in Commission Directive 84/449/EEC (which constitutes Annex V of Council Di recti ve 67/548/EEC).

A single 4-hour, semi-occluded application of the test material to the intact skin of three rabbits produced very slight erythema and very slight oedema. No skin irritation was noted 72 hours after treatment. The test material produced a primary irritation index of 0.8 and was classified as a mild irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

Conclusion

Mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals for both erythema/eschar and oedema reactions. All the reactions were fully reversible within 72 hours.

Thus, the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC) No 1272/2008.