Reaction products of disodium 3,5-diamino-2-[(2-sulfonatophenyl)diazenyl]benzoate and  diazotised sodium 2-[(4-aminophenyl)sulfonyl]ethyl sulfate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experiment start date - 25 February 2003; Experiment end date - 13 March 2003; Study completion date - 08 April 2003.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

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 (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Identification: FAT 40’810/A
Description: Brown powder
Batch number: WP 6/02
Purity: Approx. 75.0%
Stability of test item: Stable under storage conditions
Expiry date: 12-DEC-2009
Storage conditions: In the original container, at room temperature (range of 17 - 23 °C), away from direct sunlight.
Safety precautions: Routine hygienic procedures were used to ensure the health and safety of the personnel.

Specific details on test material used for the study:

Identity: FAT 40810/A
Batch: WP 6/02
Purity: approx. 75 %
Appearance: Solid, dark brownish powder
Expiration date: 12 December 2010
Storage: At room temperature at about 20 °C

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Test system: New Zealand White Rabbit, SPF
Rationale: Recognized by the international guidelines as the recommended test system.
Source: Elevage Scientifique des Dombes, F-01400 Chatillon sur Chalaronne, France
Number of animals per test: 3 (Animals of both sexes were used)
Age at treatment: 10 weeks (male), 11 weeks (females) g
Identification By unique cage number and corresponding ear number.
Acclimatization: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
Allocation: Male No. 61, Female Nos. 62 and 63

HUSBANDRY
Room no: 106/ RCC Ltd, Füllinsdorf
Conditions: Standard Laboratory Conditions, Air-conditioned with target ranges for room temperature 17 - 23 °C, relative humidity 30-70 % and approximately 10-15 air changes per hour. Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at RCC. The animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark. Music was played during the daytime light period.
Accommodation: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Füllinsdorf) and haysticks 4646 (batch no. 0206, Provimi Kliba AG) were provided for gnawing.
Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 64/02) provided by Provimi Kliba AG, CH-4303 Kaiseraugst. Results of analysis for contaminants are archived at RCC Ltd, Itingen.
Water: Community tap water from Füllinsdorf, ad libitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC Ltd, Itingen.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: The test substance was moistened with bidistilled water.

Controls:

yes, concurrent no treatment

Amount / concentration applied:

500 mg

Duration of treatment / exposure:

4 h

Observation period:

10 days

Number of animals:

3

Details on study design:

TEST ITEM PREPARATION: 0.5 g (per animal) of FAT 40810/A was weighed as delivered by the sponsor and then moistened with approximately 0.1 mL of purified water before application. The pH of the test item was measured before the study initiation date. A formulation of 1 % in water was prepared. The pH was found to be 5.19. According to Directive 92/69 EEC, B.4. and OECD Guidelines 404, a test item needs not to be tested if the pH-value is less than 2 or greater than 11.5, owing to its predictable corrosive properties.

TREATMENT: Four days before treatment, one flank was clipped with an electric clipper, exposing an area of approximately 100 sq.cm (10 cm x 10 cm). The skin of the animals was examined one day before treatment, and regrown fur of all animals was again clipped. Animals with overt signs of skin injury or marked irritation which may have interfered with the interpretation of the results were not used in the test. On the day of treatment, 0.5 g of FAT 40810/A was placed on a surgical gauze patch (ca. 4 cm x 4 cm). This gauze patch was applied to the intact skin of the clipped area. The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape. The duration of treatment was 4 hours. Then the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearly visible at that time.

OBSERVATIONS: Viability/Mortality: Daily from delivery of the animals to the termination of test. Clinical signs: Daily from delivery of the animals to the termination of test. Body weights: At start of acclimatization, on the day of application and at termination of observation.

IRRITATION SCORES: The skin reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992 approximately 1, 24, 48 and 72 hours, as well as 7, 10 and 14 days after the removal of the dressing, gauze patch and test item. To allow further examination of the test site animal No. 63 was re-clipped on completion of the 48-hour examination.

Grading of Skin Reactions
ERYTHEMA AND ESCHAR FORMATION
No erythema.............................................0
Very slight erythema..................................1
Well-defined erythema................................2
Moderate to severe erythema......................3
Severe erythema (beet redness) or eschar formation (injuries in depth preventing erythema) reading................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 (edges raised approximately 1 mm)......................................................3
Severe oedema (raised more than 1 mm and extending beyond the area of exposure)..........4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Neither oedema nor erythema were observed at any point during the study and thus there was no corrosion/irritation response observed.

Other effects:

COLORATION: The test item caused red staining of the treated skin. This effect was reversible and was no longer evident 10 days after treatment.
MORTALITY/CLINICAL SIGNS: No mortality and no clinical signs observed.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

FAT40810/A is non-irritating to skin.

Executive summary:

The primary skin irritation potential of FAT 40810/A was investigated by topical semi­ occlusive application of 0.5 g to the intact left flank of each of three young adult New Zealand White rabbits. This study was conducted in accordance with OECD test guideline 404 in a GLP certified laboratory. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours, as well as 7 and 10 days after removal of the dressing. The test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

 

The application of FAT 40'810/A to the skin resulted in no signs of irritation. The test item caused red staining of the treated skin. This effect was reversible and was no longer evident 10 days after treatment, the end of the observation period for all animals. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals.

Thus, the test item did not induce significant or irreversible damage to the skin. Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 06, 2001), FAT 40'810/A is considered to be "not irritating" to rabbit skin.