Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From May, 16 2006 to May 30, 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report date:
2006

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
475-300-1
EC Name:
-
Cas Number:
910030-59-0
Molecular formula:
Hill formula: C29H25N8Na5O22S7 CAS formula: C29H30N8O22S7.5Na
IUPAC Name:
Pentasodium 2,2'-((2,4-diamino-5-(4-methoxy-2-sulfophenylazo)-1,3-phenylene)bis(azo))bis(5-(2-(sulfonatooxyethyl)sulfonyl)benzenesulfonate
Test material form:
solid: particulate/powder
Details on test material:
see below

Test animals

Species:
rat
Strain:
Wistar
Remarks:
CRL:(WI) BR Wistar rats
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (Europe) laboratories INC.
- Hygienic level at arrival: SPF
- Age at study initiation: Young adult rats
- Body weight range at treatment: Male: 243 - 271 g; Female: 236 - 245 g
- Sex: Male and female; female rats were nulliparous and non-pregnant
- Housing: Type II polypropylene/polycarbonate, individual caging
- Bedding: Laboratory bedding
- Diet: Ssniff RIM-Z+H complete diet, ad libitum
- Water: Tap water from 500 mL bottle, ad libitum
- Acclimation period: Female: 20 d; male: 6 d
-Animal identification: Numbers on the tail written with a permanent marker; The cages were marked by identity cards with information about study code, sex, dose group, cage number and individual animal number

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3°C
- Humidity: 30 - 70%
- Air changes: 8-12 air exchanges/hour by central air-condition system
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: May, 16 2006 to May 30, 2006

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
The back of animals was shaven (approximately 10% area of the total body surface) 24 h prior to the treatment. The test substance was applied in a single dose in original form uniformly over the shaved skin throughout a 24 h exposure period. Sterile gauze pads were placed on the skin of rats. These gauzes were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was wrapped with semi occlusive plastic wrap for 24 h.

REMOVAL OF TEST SUBSTANCE
At the end of the exposure period, residual test item was removed, using body temperature water.

PREPARATION OF THE TEST SUBSTANCE
For the treatment the test substance was applied in original form moistened with 0.5 mL of distilled water.
Duration of exposure:
24 h
Doses:
2,000 mg/kg bw
No. of animals per sex per dose:
Five/sex/dose
Control animals:
not required
Details on study design:
Experimental design

Dose Group Number of animals
Male Group (2,000 mg/kg bw) 5
Female Group (2,000 mg/kg bw ) 5

A single administration was performed by dermal route and was followed by a 14 d observation period.

OBSERVATIONS

Clinical Observations

Careful clinical observation was made at the following intervals: 1h, 5h after the treatment and once each day for 14 d thereafter. Individual observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern as well. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Body Weight Measurement

The body weight of each animal was recorded on Day 0 (shortly before the treatment), then on Days 7 and 14 with a precision of 1 g.

NECROPSY

At the end of the observation period gross pathology was conducted in every experimental animal. Animals were exsanguinated under pentobarbital anaesthesia. After examination of the external appearance, the cranial, thoracic, and abdominal cavities were opened and tissues, organs were observed. Gross pathological changes were recorded for each animal on the post mortem record sheets.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No mortality occurred after the 24 h dermal exposure to test substance in CRL(WI)BR rats.
Clinical signs:
No behavioural changes or general toxic symptoms were noted during the study.

The skin remained brownish coloured after careful washing off of test substance, thus it was not feasible to evaluate erythema. In three cases (1/5 male, 2/5 female) some small wounds were observed on the treated skin surface. The skin recovered between Days 4 and 6.
Body weight:
The mean body weight and the body weight gain of animals were in the normal range during the two weeks observation period, similar to the expected values in untreated animals of the same age and strain.
Gross pathology:
Test substance related macroscopic alterations were not found at the gross pathology. Pinprick-sized haemorrhages (5/5 male, 2/5 female) were observed in the lungs, which were caused by exsanguination procedures. Hydrometra (1/5) due to the sexual cycle is a common alteration in experimental rats.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the study conditions, the dermal LD50 of the test substance was found to be >2,000 mg/kg bw in rats.
Executive summary:

A study was conducted to assess the acute dermal toxicity of the test substance in CRL:(WI) BR Wistar rats according to OECD Guideline 402, EU Method B.3. and EPA OPPTS 870.1200, in compliance with GLP.

 

Groups of five female and five male rats received a single dermal dose of 2,000 mg/kg bw. Test substance moistened with 0.5 mL of distilled water was applied topically under semi-occlusive conditions for 24 h.

 

No mortality occurred, no clinical symptoms were observed and no significant macroscopic abnormalities were seen at necropsy. Body weight was also not impaired. Although, the skin of the animals was brownish coloured by the test substance and some small wounds were observed on the treated skin surface in three animals, skin recovered between Days 4 and 6. 

Under the study conditions, the dermal LD50 of the test substance was found to be >2,000 mg/kg bw in rats