Sodium 4(-2-hydroxy-1-naphthylazo)naphthalenesulphonate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 03 Feb. 2004 to 24 Feb. 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

April 2002

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

29 DEC 1992 (92/69/EEC)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Sodium 4(-2-hydroxy-1-naphthylazo)naphthalenesulphonate

EC Number:

216-760-3

EC Name:

Sodium 4(-2-hydroxy-1-naphthylazo)naphthalenesulphonate

Cas Number:

1658-56-6

Molecular formula:

C20H13N2NaO4S

IUPAC Name:

NA

Test material form:

solid: particulate/powder

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Granja Cunicola San bernardo, S.L
- Age at study initiation: 9-10 weeks old
- Weight at study initiation: 2.4 - 2.7 kg
- Housing: individually in stainless steel cages (52.0 x 58.0 X 43.0 cm or 45.0 x 59.0 X 37.0 cm) with a grille floor placed on racks.
- Diet: free access to a standard rabbit diet, Panlab 112, (Panlab, S.L., Barcelona, Spain, expiry date: 25.Mar.04, 08.Apr.04 and 10.june.04) which had been analysed by the manufacturer.
- Water: supplied ad libitum, by means of an automatic drinking system or in bottles. The tap water, supplied by Compaña de Aguas de Sabadell, S.A., is analysed periodically for contaminants.
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
-Temperature: 19 - 23 °C
-Humidity: 30 - 60 %
-Photoperiod: 12 hours cycle dark/light (7:00 am to 7:00 pm)

Test system

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g moistened with distilled water before the administration

Duration of treatment / exposure:

3 minutes, 1 hour and 4 hours in one animal; 4 hours in the other two animals

Observation period:

14 days

Number of animals:

3 rabbits

Details on study design:

TEST SITE
- Area of exposure: the animals were shaved 24 hours before administration using an electric razor. One square test zone, measuring approximately 6.2 cm², was delimited on the shaven intact skin of one side of each animal.
- % coverage: 2.5 x 2.5 cm
- Type of wrap: patches were held close to the body using a microporous and hypoallergenic adhesive tape; a strip of gauze was wrapped around the trunk of the animal and fixed in place with hypoallergenic adhesive tape in order to hold the semiocclusive patch in position.

TEST ITEM ADMINISTRATION
At first only one animal was used, applying three patches simultaneously for 3 minutes, 1 and 4 hours. After each one of these exposure periods, the corresponding bandage and patch were removed.
The test was completed using two other animals, applying a dressing to each one for 4 hours. After this exposure period, the bandages and patches were removed.

REMOVAL OF TEST SUBSTANCE
- Washing: with water and PEG 400
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
1, 24, 48, 72 hours, 7 and 14 days

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

Edema Formation
No edema: 0
Very slight edema (barely perceptible): 1
Slight edema (edges of area well defined by definite raising): 2
Moderate edema (raised approximately 1 mm): 3
Severe edema (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, #2, #3

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 7 days

Results 2

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48 h

Results 3

Irritation parameter:

edema score

Basis:

animal: #2, #3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Initially, three dressing were applied simultaneously to one animal. The first was removed after 3 minutes, the second after 1 hour and the thirs after 4 hours of exposure.
No skin lesions were recorded in the application area, 1 hour after removing the first semiocclusive patch, which was exposed for 3 minutes. No lesions were observed either 24, 48, 72 hours, 7 and 14 days after removing the patch.
Sixty minutes after removal of the patch exposed for 1 hour, well-defined erythema (grade 2) with very slight edema (grade 1) was recorded. Twenty-four hours after removing the patch, the edema remitted completely, and the erythema decreased to very slight erythema (grade 1), which persisted until 48 hours after removing the patch.
No skin lesions were recorded 72 hours, 7 and 14 days after the removal of the patch exposed for 1 hour.
Sixty minutes after removing the patch exposed for 4 hours, well-defined erythema (grade 2), accompanied by very slight edema (grade 1), was observed. After 24 hours, very slight edema (grade 1) persisted, and well-defined erythema (grade 2) developed into very slight erythema (grade 1).
During the observations carried out 48 and 72 hours afier removing the patch, the edema had remitted completely although very slight erythema persisted (grade 1).
No skin alterations were recorded 7 and 14 days after the removal of the patch exposed for 4 hours.
Subsequently, two more animals were used and a dressing with the test item, which was removed after a four-hour exposure, was applied to them.
Sixty minutes after removing the patch exposed for 4 hours, both animals presented well-defined erythema (grade 2), accompanied by very slight edema (grade 1) in one of them, and slight edema (grade 2) in the other one.
Twenty-four, 48 and 72 hours from the removal of the patch, very slight erythema (grade 1) was recorded in both animals.
No skin lesions were recorded in any of the two animals in the observations made 7 and 14 days after removing the patch exposed for 4 hours.

Other effects:

Clinical signs: the behaviour and physical state of the rabbits were normal throughout the Study.

Any other information on results incl. tables

Overall test results

Aninmal N. (time)	Reaction	60 min	24 hrs	48 hrs	72 hrs	7 days	14 days	Mean 24/48/72
# 1	Erythema	2	1	1	1	0	0	1.00
# 2	Erythema	2	1	1	1	0	0	1.00
# 3	Erythema	2	1	1	1	0	0	1.00
# 1	Edema	1	1	0	0	0	0	0.33
# 2	Edema	1	0	0	0	0	0	0.00
# 3	Edema	2	0	0	0	0	0	0.00

Reactions animal 1

Aninmal N. (time)	Reaction	60 min	24 hrs	48 hrs	72 hrs	7 days	14 days
1 (3 min)	Erythema	0	0	0	0	0	0
1 (1 hr)	Erythema	2	1	1	0	0	0
1 (4 hrs)	Erythema	2	1	1	1	0	0
1 (3 min)	Edema	0	0	0	0	0	0
1 (1 hr)	Edema	1	0	0	0	0	0
1 (4 hrs)	Edema	1	1	0	0	0	0

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

The substance was found not to be not irritating to the rabbit skin.

Executive summary:

The degree of primary skin irritation caused by the test item was evaluate in a study performed according to the OECD Guideline No. 404 (2002) and the Method B.4 of European Communities Commission Directive of 31 July 1992 (92/69/EEC). Initially three semiocclusive patches were applied with 0.5 g of test item simultaneously to one animal for 3 minutes, 1 and 4 hours, respectively. A semiocclusive patch with 0.5 g of test item was applied for 4 hours in the other two animals. The observation time points were at 1, 24, 48, 72 hours and 7, 14 days after the administration.

In all animals the mean scores (24/48/72 h) for erythema were 1, while for oedema the mean value was 0.33 in one animal (#1) and 0 in the other two animals (#2 and #3). No skin lesions were recorded in any of the animals during the observations made 7 and 14 days after removing the patch.