Tetrasodium 3,3'-[[6-[(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diyl]bis[imino(2-methyl-4,1-phenylene)azo]]bisnaphthalene-1,5-disulphonate

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:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

500 mg of the pulverized test substance were moistened with water (to ensure good contact with the skin) and subsequently applied to a hypoallergenic Hansamed® - patch. A further patch was moistened with water. The patches prepared in this way were placed on the opposite dorso-lateral areas of the trunk of each animal and were loosely held in place with a semiocclusive dressing for the duration of the exposure period. The treated skin area was approx. 6 cm² in size. After an exposure period of four hours, the dressing and patches were removed. The exposed skin areas were carefully washed with water and scored according to Draize.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Tetrasodium 3,3'-[[6-[(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diyl]bis[imino(2-methyl-4,1-phenylene)azo]]bisnaphthalene-1,5-disulphonate

EC Number:

256-851-5

EC Name:

Tetrasodium 3,3'-[[6-[(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diyl]bis[imino(2-methyl-4,1-phenylene)azo]]bisnaphthalene-1,5-disulphonate

Cas Number:

50925-42-3

Molecular formula:

C39H30N10Na4O13S4

IUPAC Name:

tetrasodium 3,3'-({6-[(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diyl}bis[imino(2-methyl-4,1-phenylene)diazene-2,1-diyl])dinaphthalene-1,5-disulfonate

Specific details on test material used for the study:

State of Aggregation: solid
External Appearance: yellow-brown powder
contents: 100%
pH: 6-7 (40 g/L H2O)

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species and species justification
The study was conducted on rabbits an animal species recommended in the guidelines for this type of study.
Healthy adult albino rabbits, strain HC:NZW were used. The health of the animals was routinely examined for the main specific pathogens by the breeder. No vaccinations or treatment with antibiotics were perf'ormed prior to receipt of' the animals or during the acclimatization phase or study period.

Housing and feeding conditions
The rabbits were individually housed in stainless steel cages with flat rod bases or plastic cages with perforated bases, under standardized conventional conditions. Excrement trays beneath the cages contained low-dust (wood) bedding. Bedding was regularly spot-checked for contaminants at the instance of the Department of Laboratory Animal Services, and changed at least twice weekly.
Identification of animals: The rabbits were identified by individual ear marks (tattoos) and cage cards.

Acclimatization:
Prior to the initiation of the treatment the animals were kept for at least 14 days in the quarantine station of the Department of Laboratory Animal Services and monitored for diseases. During this period pooled faeces specimens were examined for Coccidia oocysts.

Animal housing conditions:
All the animals in this study were kept in one room.

Climatic conditions in animal room:
The environmental conditions were adjusted as follows:
Room temperature: 20 ± 3 °C
Humidity, relative: approx. 50 %
Light-/Dark cycle: 12 hours, artificial illumination
Air exchange rate: approx. 10 times per hour

Nutrition:
Feed: Standard diet "Ssniff K 411 (Ssniff Spezialdiäten GmbH, Soest), approx. 100 - 120 g per animal/day; once per day in the morning.
Water: Tap water; ad libitum.
Drink-water was supplied either in polycarbonate bottles containing approx. 750 ml or from automatic watering.

Weight of animals
The animals were weighed immediately before application of the test substance.

Randomization
Rabbits were randomly assigned to the respective treatment groups.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

other: shorn

Vehicle:

water

Controls:

not required

Amount / concentration applied:

500 mg of the pulverized test substance

Duration of treatment / exposure:

4 hours

Observation period:

7 days

Number of animals:

3 animals

Details on study design:

Test for irritant/corrosive effects - skin

Procedure
Approximately 24 hours before the test, fur was shorn from the dorso-lateral area of the trunk (6 x 6 cm) of each of three rabbits. 500 mg of the pulverized test substance were moistened with water (to ensure good contact with the skin) and subsequently applied to a hypoallergenic patch. A further patch was moistened with water. The patches prepared in this way were placed on the opposite dorso-lateral areas of the trunk of each animal and were loosely held in place with a semiocclusive dressing for the duration of the exposure period. Thus, access by the animal to the patch and resultant ingestion/inhalation of the test substance was prevented. The treated skin area was approx. 6 cm² in size. After an exposure period of four hours, the dressing and patches were removed. The exposed skin areas were carefully washed with water without altering the existing response, or the integrity of the epidermis. The contralateral skin area not treated with test substance served as control.

Clinical observations and scoring
Dermal irritation was scored and recorded after termination of exposure at 1h, 24h, 48h, 72h and 7d . The degree of erythema/eschar formation and oedema formation was recorded as specified by DRAIZE, and any serious lesions or toxic effects other than dermal irritation were also recorded.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Remarks on result:

other: evaluation not possible due to the intense coloration by the test substance

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: score = 0 at any time point

Irritant / corrosive response data:

During the first 72 hours evaluation of erythema was not possible due to the intense coloration by the test substance (exposure period: 4 hrs). Other dermal reactions (e.g. edema) were not observed during this period and also no skin erythema became evident on day 7. Therefore, a significant irritant potential of the test substance to the skin is unlikely.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Evaluation of erythema was not possible during the first 72 hours due to the intense coloration by the test substance (exposure period: 4 hrs). Edema score was = at any time point (1h, 24h, 48h, 72h and 7d).

Executive summary:

500 mg of the pulverized test substance were moistened with water (to ensure good contact with the skin) and subsequently applied to a hypoallergenic Hansamed® - patch. ( A further patch was moistened with water. The patches prepared in this way were placed on the opposite dorso-lateral areas of the trunk of each animal and were loosely held in place with a semiocclusive dressing for the duration of the exposure period. The treated skin area was approx. 6 cm² in size. After an exposure period of four hours, the dressing and patches were removed. The exposed skin areas were carefully washed with water and scored according to Draize. During the first 72 hours evaluation of erythema was not possible due to the intense coloration by the test substance (exposure period: 4 hrs). Other dermal reactions  (e.g. edema) were not observed during this period and also no skin erythema became evident on day 7. Therefore, a significant irritant potential of the test substance to the skin is unlikely.