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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experiment start date - 06 February 2003; Experiment end date - 27 February 2003; Study completion date - 13 March 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 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Identity: FAT 40’810/A
Description: Brown solid
Batch number: WP 6/02
Expiry date: 12-DEC-2009
Purity: approx. 75.0%
Stability of test item dilution: For at least 7 days in water at room temperature.
Storage conditions: In the original container, at room temperature (range of 20 ±3 °C, provided at RCC Ltd), away from direct sunlight.

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:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test system: Rat, HanBrl: Wist (SPF)
Rationale: Recognized by the international guidelines as a recommended test system.
Source: RCC Ltd, Biotechnology and Animal Breeding, CH-4414 Füllinsdorf / Switzerland
Number of animals per group 3 males and 3 females
Total number of animals 3 males and 3 females
Age when treated: Males: 8 weeks, Females: 12 weeks
Identification: Unique cage number and corresponding color-coded spots on the tail. The animals were marked immediately prior to treatment.
Randomization: Selected by hand at time of delivery. No computer generated randomization program.
Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.
Husbandry: Room no. 104/ RCC Ltd, Füillinsdorf
Conditions: Standard Laboratory Conditions. Air-conditioned with 10 – 15 air changes per hour, and continuously monitored environment with target ranges for temperature 22 ± 3 °C and for relative humidity between 30 - 70 % (values above 70 % during cleaning process possible), 12 hours fluorescent light 12 hours dark, music during the light period.
Accommodation: In groups of three per sex in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).
Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 67/02 (Provimi Kliba AG, CH-4303 Kaiseraugst Switzerland) ad Iibitum. Results of analyses for contaminants are archived at RCC Ltd, ltingen.
Water: Community tap water from Füllinsdorf ad Iibitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC Ltd, ltingen.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

Bi-distilled

Details on dermal exposure:

One day before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 10 % of the total body surface. Only those animals without injury or irritation on the skin were used in the test. On test day 1, the test item was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe and covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage (application volume/kg body weight: 6 ml). Twenty-four hours after the application the dressing was removed and the skin was flushed with lukewarm tap water and dried with disposable paper towels. Thereafter, the reaction sites were assessed.

Duration of exposure:

24 h

Doses:

2000 mg /kg bw

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

Observations:
Mortality / Viability: Daily during acclimatization and twice daily during days 1-15.
Body weights: On test days 1 (prior to administration), 8 and 15.
Clinical signs: Daily during acclimatization and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15.
Necropsy: All animals were killed at the end of the observation period by an intraperitoneal injection of Vetanarcol at a dose of at least 2.0 ml/kg body weight (equivalent to at least 324 mg sodium pentobarbitone/kg body weight) and discarded after macroscopic examinations were performed. No organs or tissues were retained.

Statistics:

No statistical analysis was used.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Male: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0
Female: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0

Clinical signs:

other: No clinical signs were observed during the observation period. Slight red staining of the treated skin area was observed in all animals immediately after removal of the dressing and persisted up to the end of the observation period.

Gross pathology:

Effects on organs: No macroscopic findings observed at necropsy.

Other findings:

Slight red staining of the treated skin area was observed in all animals immediately after removal of the dressing and persisted up to the end of the observation period.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

LD50 (dermal, rat): >2000 mg/kg bw

Executive summary:

In a GLP-study conducted according to OECD test guideline 402, FAT 40810/A was assessed for its toxicity potential upon single application to skin. Five male and five female HanBrI: WIST (SPF) rats were treated with FAT 40810/A at 2000 mg/kg by dermal application. The test item was diluted in vehicle (purified water) at a concentration of 0.33 g/ml and administered at a volume dosage of 6 ml/kg. The application period was 24 hours. No deaths occurred during the study. No clinical signs were observed during the observation period. Slight red staining of the treated skin area was observed in all animals immediately after removal of the dressing and persisted up to the end of the observation period. Three female animals showed a loss of body weight (0.3 % to 6.2%) one week after treatment whereas one other female animal did not gain body weight during the same period. They recovered at the end of the observation period. The body weight of all other animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy except the slight red staining of the treated skin area which was still present. In conclusion the median lethal dose of FAT 40810/A after single dermal administration to rats of both sexes, observed over a period of 14 days is LD50 (rat):greater than 2000 mglkg body weight.