Reaction mass of 3,3'-(1E,1'E)-(4,4'-(6-(bis(2-hydroxyethyl)amino)-1,3,5-triazine-2,4-diyl)bis(azanediyl)bis(3-methoxy-4,1-phenylene))bis(diazene-2,1-diyl)dibenzenesulfonic acid and disodium 3,3'-[[6-[bis(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diyl]bis[imino(3-methoxy-4,1-phenylene)azo]]bis[benzenesulphonate]

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

other: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From May 09 to 12, 2006

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Remarks:

Source study has reliability 1. Details on the read across are attached in section 13.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 24 April 2002

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:

Similar Substance 01

IUPAC Name:

Similar Substance 01

Test material form:

not specified

Specific details on test material used for the study:

Test material was used as supplied.
pH of test material was determined before starting the study and found to be 9.1 at 10 % w/w aqueous preparation of test material.

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, UK.
- Weight at study initiation: 2.0 to 3.5 kg
- Housing: the animals were individually housed in suspended cages.
- Diet: ad libitum.
- Water: drinking water, ad libitum.
- Acclimation period: at least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature: 17 - 23 °C
- Humidity: 30 - 70 %
- Air changes: the rate of air exchange was at least fifteen changes per hour.
- Photoperiod: the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 g of test material, moistened with 0.5 ml of distilled water

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

three rabbits

Details on study design:

TEST SITE
- Area of exposure: on the day before the test each 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.
- Application: test material was introduced under a 2.5 cm × 2.5 cm cotton garze 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. To prevent the animals interfering with patches, the trunk of each rabbit was wrapped in an elasticated corset 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.
- Time after start of exposure: 4 hours.

OBSERVATION TIME POINTS
Approximately one hour following the removal of the patches and 24,48 and 72 hours later, test sites were examined for evidence of primary irritation and scored.
Scores for erythema and oedema at 24 and 72-hour readings were based on the scheme devised by Draize J H (1959) "Dermal Toxicity" In: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. Association of Food and Drug Officials of the United States, Austin, Texas, p.47.

SCORING SYSTEM:
Erythema and Eschar formation
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) to eschar formation preventing grading of erythema 4

Oedema Formation
No oedema 0
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

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Yellow-coloured staining was noted at all treated skin sites throughout the study. This did not affect evaluation of skin reactions. No evidence of skin irritation was noted during the study.
The test material produced a primary irritation index of 0.0. No corrosive effects were noted.

Any other information on results incl. tables

Individual skin reactions

Animal	Skin Reaction	Time				Mean 24/48/72 hrs
		1 hr	24 hrs	48 hrs	72 hrs
Male 155	Erythema/Eschar	0 STA	0 STA	0 STA	0 STA	0 STA
Male 156	Erythema/Eschar	0 STA	0 STA	0 STA	0 STA	0 STA
Male 157	Erythema/Eschar	0 STA	0 STA	0 STA	0 STA	0 STA
Male 155	Oedema	0	0	0	0	0
Male 156	Oedema	0	0	0	0	0
Male 157	Oedema	0	0	0	0	0

STA : Yellow-coloured staining

Sum of 24 and 72-hour readings (S): 0

Primary Irritation Index: 0.0

Classification: non-irritant

Applicant's summary and conclusion

Interpretation of results:

other: not classified, according to the CLP Regulation (EC 1272/2008)

Conclusions:

Not skin irritant.

Executive summary:

The study was performed to assess the irritancy potential of test material to the skin of New Zealand White rabbit, following OECD guideline 404 and EU method B4.

A single 4 -hour, semi-occlusive application of test material to the intact skin of three rabbits produced no evidence of skin irritation. Mean values from gradings at 24, 48 and 72 h after patch removal were 0 for both erythema and oedema and primary irritation index was 0.0, according to the Draize classification scheme.