A mixture of: disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(ethensulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex; disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(2-hydroxyethylsulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07 November 1995 to 10 November 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

A mixture of: disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(ethensulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex; disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(2-hydroxyethylsulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex;

EC Number:

423-940-7

EC Name:

A mixture of: disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(ethensulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex; disodium 6-[3-carboxy-4,5-dihydro-5-oxo-4-sulfonatophenyl)pyrazolin-4-yl-azo]-3-[2-oxido-4-(2-hydroxyethylsulfonyl)-5-methoxyphenylazo]-4-oxidonaphthalene-2-sulfonate copper (II) complex;

Cas Number:

85585-91-7

IUPAC Name:

4-[2-(7-{2-[4-(ethenesulfonyl)-2-hydroxy-5-methoxyphenyl]diazen-1-yl}-8-hydroxy-6-sulfonaphthalen-2-yl)diazen-1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid 4-[2-(8-hydroxy-7-{2-[2-hydroxy-4-(2-hydroxyethanesulfonyl)-5-methoxyphenyl]diazen-1-yl}-6-sulfonaphthalen-2-yl)diazen-1-yl]-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazole-3-carboxylic acid dicopper tetrasodium hydride

Test material form:

solid

Specific details on test material used for the study:

Identification: Pacified Reactive Black 31
Description: Dark blue crystals
Batch: 9T-55
Purity: 89%
Storage: At room temperature in the dark
Stability in vehicle: Stable in water for at least 96 hours

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Age at start of treatment
Approximately 8 0r 9 weeks

Body weight at start of treatment
1532 - 1691 grams

Identification
Ear tag

Conditions
Air-conditioned room with approximately 15 air changes per hour and the environment controlled with optimal conditions considered as being a temperature of 21°C and a relative humidity of 50%. Fluctuations from these optimal conditions were noted, but were considered not to have affected study integrity. Lighting was 12 hours artifical fluorescent light and 12 hours dark per day.

Accomodation
Individually in labelled cages with protected floors and equipped with an automatic drinking system. Acclimatisation oeruid was at least 5 days before start of treatment under laboratory conditions.

Diet
Standard laboratory rabbit diet, approximately 100 g per day. Certificates of analysis were examined and retained in the NOTOX archives. In addition, hay was provided once a week.

Water
Free access to tap water diluted with decalcified water.

Certificates if analysis were examined and retained in the NOTOX archives..

Test system

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 gram of mositeed test material

Duration of treatment / exposure:

Four hours

Observation period:

72 hours

Number of animals:

3 male rabbits

Details on study design:

Approximately 24 hours before treatment, the dorsal fur was shaved with electric clippers, exposing an area of approximately 150 cm2.

the animals were examined, and the skin to be treated in particular, prior to test substance administration and no abnormalities were detected.

On test Day 1, 0.5 g if the test substance was moistened and subsequently applied to te intacj skin if the shaved area on one flank. A similar patch, but without test substance and water, was applied to the contralateral flank, to act as a procedural control. Both patches were mounted on tape, which was wrapped around the abdomen and secured with an elastic bandage.

Whenever considered necessary, the skin areas concerned of the animals we re-shaved at leat 3 hours before the observations, to facilitate the scoring.

Observations
Mortality/viability
Twice daily

Toxicity
At least once daily

Body weight
Day of treatment (prior to application)

Irritation
The skins reactions were assessed at approximately 1, 24, 48 and 72 hours after removal of the dressings and test substance. The irritation scores and a description of all other (local) effects were recorded.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Noeffects observed

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Irritant / corrosive response data:

No skin irritation was caused by Pacified Reactive Black 31 after 4 hours of exposure in two animals. Very slight erythema was noted in the treated skin-area if the third animal, one hour after exposure only.

Other effects:

Colouration
Black staining of the treated skin by the test substance was observed in all animals during the observation period.

Any other information on results incl. tables

Toxicity symptoms

No symptoms of systemic toxicity were oberved in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Dermal application of Pacified Reactive Black 31 resulted in a primary irritation index of 0 (non-irritating) when applied to the intact rabbit skin

Executive summary:

The study was carried out in accordance with the OECD Guideline No. 404, "Acute Dermal Irritation / Corrosion" and the EEC Directive 92/69/EEC, B.4 "Acute Toxicity - Skin Irritation".

Three rabbits were exposed to 0.5 g of Pacified Reactive Black 31, applied onto shaved skin for 4 hours using a semi-occlusive dressing. Observations were made 1, 24, 48 and 72 hours after exposure.

Exposure to Pacified Reactive Black 31 resulted in a very slight erythema in the treated skin area in one animal, which had resolved within 24 hours after expsoure. No skin irritation was observed in the other two animals.

Black staining of the treated skin by Pacified Reactive Black 31 was observed during the observatin period.

No signs of systmic toxicity were observed udring the study period.

Dermal application of Pacified Reactive Black 31 resulted in a primary irritation index of 0 (non-irritating) when applied to the intact rabbit skin.