2,7-Naphthalenedisulfonic acid, 4-amino-6-[2-[4-[[[4-[2-(2,4-dihydroxyphenyl)diazenyl]phenyl]sulfonyl]amino]phenyl]diazenyl]-5-hydroxy-3-[2-(4-nitrophenyl)diazenyl]-, sodium salt (1:2)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

other: read across from supporting substance

Adequacy of study:

key study

Study period:

1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The test has been performed following official guidelines, that completely assesses the end point, on a similar substance differing only by the counter ion potassium instead of sodium cation.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4-amino-6-[[4-[[[4-[(2,4-diaminophenyl)azo]phenyl]amino]sulphonyl]phenyl]azo]-5-hydroxy-3-[(4-nitrophenyl)azo]naphthalene-2,7-disulphonic acid, potassium salt

EC Number:

286-384-2

EC Name:

4-amino-6-[[4-[[[4-[(2,4-diaminophenyl)azo]phenyl]amino]sulphonyl]phenyl]azo]-5-hydroxy-3-[(4-nitrophenyl)azo]naphthalene-2,7-disulphonic acid, potassium salt

IUPAC Name:

286-384-2

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

A total of 3 New Zeland White male rabbits were supplied by B&K Universal GJsl. On arrival at Centro de Investigacion Aplicado SAL the animals weighed 2.0-2.1 kg and were aged approxiamtely 9-10 weeks.
Before administration they were subjected to a prelimianry observation and acclimtization period of at least 7 days. At the start of the study the animals were in perfect health codnitions and weighed 2.2-2.3 kg.

Identification
The rabbits were individually identified using numbered ear tags. The number assigned to each rabbit was stated on the identification card on the cage.

Group size
The substance was tested on one group of three animals. The numbers assigned to the animals were 167, 184 and 195.

Housing
The rabbits were housed individually in stainless steel cages (48x59x37 cm) with a grille floor, placed on racks.
Each cage was identified by a card stating the study code number, administration route, animal number and sex, name or code of the test substance , animal delivery and administration date and study director's name..

Temperature in the animal room was maintained at 17-21°C. Relavtive humidity was between 45-70%.

Lighting was controlled to provide 12 hours light (7:00 to 19:00 hiours) and 12 hours dark per 24 hour period.

The animals were allowed free access to s standard rabbit diet UAR 112 which had been analysed by the manufacturer.

Water was supplied ad libitum by an automatic drinking system. The water, supplied by "Compania de Aguas de Sabadell SA" is periodically checked for the possble presence of contaminants.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

other: dampened with water

Controls:

other: the untreated skin areas of the test animal serve as the control

Amount / concentration applied:

0.5g placed on a 2.5 x 2.5 cm squares of surgical gauze and applied to the corresponding test area

Duration of treatment / exposure:

4 hours

Observation period:

The reaction of the test zone was evaluated approximately 60 minutes, 24, 48 and 72 hours after the end if the exposure period.

Number of animals:

3 male rabbits

Details on study design:

The animals were shaved approximately 24 hours before administration usign an electric razor, leaving the back and sides exposed from the scapula to the pelvis. One square test zone measuring approximately 6.2 cm squares was delimited on the shaved intatc skin of one side of the animals. The substance was placed on 2.5 x 2.5 cm squares of srugical gauze and applied to the patches in place. The resulting patches were held close to the body using strips of adhesive tape. Finally, a strip of gauze was fixed to the body using hypoallergenic adhesive tape to maintain the semi-occlòusive bandage in place. The aim of this semi-occlusive bandage was to allow good skin absorption of the test substance. Following administration, the animals were observed for a few moments before being returned to their cages. Four hours later, the bandage and patches were removed and the remains of the product were removed with distilled water.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 60 min

Score:

ca. 1

Results 2

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 24h, 48h, 72h

Score:

0

Results 3

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 60min, 24h, 48h, 72h

Score:

0

Results 4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 60 min

Score:

ca. 1

Results 5

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 24h, 48h, 72h

Score:

0

Results 6

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 60 min

Score:

0

Results 7

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24h, 48h, 72h

Score:

0

Results 8

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 60min, 24h, 48h, 72h

Score:

0

Results 9

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 60min, 24h, 48h, 72h

Score:

0

Other effects:

The behaviour and physical condition of the rabbits were normal throughout the study

Applicant's summary and conclusion

Interpretation of results:

other: not irritant

Conclusions:

The substance was administered for the study of primary skin irritation in rabbits. After assessment of the alterations caused according to OECD 404 and EU B4, it is considered NON IRRITANT to the skin.

Executive summary:

The degree of skin irritation caused by substance was determined after it was applied to shaved rabbit skin, under semi-occlusive patch and left for 4 hours. After assessment and evaluation of the alterations which it induced according to OECD 404 and EU B4 it is considered NON IRRITANT. In the examinations carried out in the first 60 minutes after removing the semi-occlusive patch, two animals presented very slight erythema (grade 1). In the examinations carried out 24, 48 and 72 hours after treatment, no dermal lesions were recorded in any of the animals. The mean erythem and edema scores for the substance at 24, 48 and 72 hours in the three animals, were both 0.