Reaction products of diazotised 2-amin-6-nitrophenol-4-sulphonic Acid and 2-Amino-4-nitrophenol-6-sulfonic Acid, coupled with a mixture of 6-Amino-4-hydroxy-2-naphthalenesulfonic Acid and Acetylated 6-Amino-4-hydroxy-2-naphthalenesulfonic Acid, subsequently metallized with Chromium (III) Basic Sulfate and Acetic Acid, and at the end diazotised and coupled with p-Cresol, sodium salts

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

May 12, 1981

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

405-750-6

EC Name:

-

Molecular formula:

Not applicable for UVCB substance

IUPAC Name:

diiron(3+) dihydrogen tris(7-[2-(2-hydroxy-5-methylphenyl)diazen-1-yl]-2-[2-(3-nitro-2-oxido-5-sulfophenyl)diazen-1-yl]-3-sulfonaphthalen-1-olate) 7-acetamido-2-[2-(3-nitro-2-oxido-5-sulfophenyl)diazen-1-yl]-3-sulfonaphthalen-1-olate

Test animals

Species:

rabbit

Strain:

New Zealand White

Remarks:

SPF-Quality

Details on test animals or test system and environmental conditions:

TEST ANIMALS
-Source: Broekman Institute, Someren / The Netherlands
-Age at start of treatment: approx. 13 weeks
-Body weight at start of treatment: 2515-2882 g
-Identification: ear tag
-Acclimatisation: at least five days under test conditions
-Housing: individual cages with perforated floors equipped with an automatic drinking system (ITL, Bergen / The Netherlands).
-Diet: standard laboratory rabbit diet (LKK-20, pellet diameter 4mm, Hope Farms, Woerden / The Netherlands), approx. 100 g per day
-Water: tap-water diluted with decalcified water, ad libitum.

ENVIRONMENTAL CONDITIONS
-Temperature: 21 ± 3°C
-Humidity: 30-70 %
-Air changes: 7.5-15 air changes per hr with air-conditioned
-Photoperiod: 12 hrs artificial fluorescent light / 12 hrs dark.

Test system

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

milli-RO water (water prepared by reverse osmosis)

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
-Amount applied: 0.5 g/animal
-Preparation: moistened with milli-RO water (water prepared by reverse osmosis)

Duration of treatment / exposure:

4 hours of application

Observation period:

40 min, 24, 48 and 72 h after the removal of the dressings

Number of animals:

3 females

Details on study design:

TREATMENT
Approximately 24 h before treatment, the dorsal fur was shaved with an electric clipper, exposing an area of approximately 100 cm2 (10 cm * 10 cm).
Prior to test substance administration, the animals were physically examined and the skin found to be intact and normal.
The test substance was applied to the intact skin of the shaved area, using a Metalline patch (2 * 3 cm) mounted to Micropore tape. The dressing was wrapped around the abdomen and anchored with an elastic bandage.

REMOVAL OF TEST SUBSTANCE
The remaining test substance was removed using a dry tissue and subsequently a tissue moistened with tap-water.

OTHER OBSERVATIONS
-Viability / Mortality, daily.
-Body Weight, once (application day)
-Staining of the treated skin area related to the test substance was described and recorded during each observation.

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 (beet redness) to slight eschar formation (injuries in depth): 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 mm): 3
Severe oedema (raised more than 1 mm and extending beyond area of exposure): 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 24 h

Results 2

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 3

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 h

Results 4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 5

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 24 h

Results 6

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

IRRITATION
The irritation was reversible within 24 h after exposure in two animals and within 48 h in the third animal.
CORROSION
No corrosive effect was evident on the skin.

Other effects:

TOXICITY SYMPTOMS / MORTALITY
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.
COLOURATION
In the area of application black staining of the treated skin by the test substance was observed.

Applicant's summary and conclusion

Interpretation of results:

other: Not classified according to CLP Regulation (EC 1272/2008)

Conclusions:

The test substance was found to be non irritant when applied to intact rabbit skin.

Executive summary:

The skin irritation test was performed to rabbit skin according to the OECD guideline 404 (1981). The observed skin irritation consisted of very slight erythema (grade 1) in all animals. The irritation was reversible within 24 hours after exposure in two animals and within 48 hours in the third animal. In the area of application, black staining of the treated skin by test substance was observed. No corrosive effect occurred on the skin and no signs of systemic intoxication were observed in any of the rabbits.