Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
acute toxicity: dermal
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:
1989
Report date:
1989

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
February 24, 1987
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
405-750-6
EC Name:
-
Molecular formula:
Not applicable for UVCB substance
IUPAC Name:
Reaction products of diazotised 2-amin-6-nitrophenol-4-sulphonic Acid and 2-Amino-4-nitrophenol-6-sulfonic Acid, coupled with a mixture of 6-Amino-4-hydroxy-2-naphthalenesulfonic Acid and Acetylated 6-Amino-4-hydroxy-2-naphthalenesulfonic Acid, subsequently metallized with Chromium (III) Basic Sulfate and Acetic Acid, and at the end diazotised and coupled with p-Cresol, sodium salts

Test animals

Species:
rat
Strain:
Wistar
Remarks:
albino
Sex:
male/female
Details on test animals or test system and environmental conditions:
-Source: Charles River Wiga, Sulzfeld, West-Germany
-Age at start of treatment: approx. 7 weeks
-Body weight at start of treatment: males 261-298 g, females: 178-206 g
-Identification: earmark
-Acclimation period: at least 5 days under laboratory conditions
-Housing: groups of five per sex in polycarbonate cages containing purified sawdust as bedding material (Woody Clean supplied by Broekman Institute, Someren, The Netherlands).
-Diet: standard pelleted laboratory animal diet (RMH-B from Hope Farms, Woerden, The Netherlands), ad libitum
-Water: tap-water, ad libitum.

ENVIRONMENTAL CONDITIONS
-Temperature: 21 ± 3 °C
-Humidity: 30-70 %
-Air changes: 7.5-15 per hr with air-conditioned
-Photoperiod: 12 hrs artificial fluorescent light / 12 hrs dark per day.

Administration / exposure

Type of coverage:
occlusive
Vehicle:
water
Remarks:
prepared by reverse osmosis
Details on dermal exposure:
TREATMENT
-Method: dermal application
-Frequency: once, on day 1
-Dose volume: 10 ml/kg bw.

TEST SITE
-Shaving: an area of approximately 5x7 cm on the back of the animal was clipped one day before treatment (day -1)
-Area of exposure: approximately 25 cm2 (5x5 cm) for males, 18 cm2 (3.5x5 cm) for females
-Type of wrap if used: gauze patch fixed successively to aluminium foil and flexible bandage (Coban, 3M, St. Paul, U.S.A.) with drops of petrolatum.

REMOVAL OF TEST SUBSTANCE
Residual test substance was removed with tissue moistened with tap-water.
Duration of exposure:
24 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 males at 2000 mg/kg bw
5 females at 2000 mg/kg bw
Control animals:
no
Details on study design:
TEST SUBSTANCE PREPARATION
The formulations were prepared immediately prior to dosing. The test substance was weighed into a glass flask on an analytical balance and the vehicle (w/w) was added. Homogeneity was obtained by a homogeniser.
-Concentration in vehicle: varied to allow constant dosage volume in terms of ml/kg body weight.

OBSERVATIONS
-Mortality/Viability: at periodic intervals on the day of dosing (day 1) and twice daily thereafter for at least 14 days.
-Body Weights: test days 1 (pre-administration), 8 and 15.
-Symptoms: at periodic intervals on day 1 and once daily thereafter. All signs of reaction to treatment were recorded with particular attention paid to changes in the skin, fur, eyes and mucous membranes, as well as to behaviour pattern, tremors, convulsion, salivations, diarrhoea, lethargy, sleep and coma.
-Skin irritation: changes of the treated skin were described immediately after bandage removal (day 2) and on days 5, 8 and 15.

NECROPSY FINDINGS
All animals were necropsied and descriptions of all macroscopic abnormalities were recorded. All animals surviving to the end of the observation period were sacrificed by carbon dioxide asphyxiation and subjected to necropsy.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during the study period.
Clinical signs:
All animals were noted as lethargic on day 2.
No other clinical signs of toxicity or behavioural changes were seen during the study.

Body weight:
All animals showed body weight gain during the study period.
Gross pathology:
Macroscopic examination of all animals at the end of the study did not reveal any abnormalities.
Other findings:
No signs of irritation were seen on the treated skin. However, the treated skin surface of all the animals was discoloured black on days 2 and 5, due to staining by the test substance, which made a detailed examination difficult.

Applicant's summary and conclusion

Interpretation of results:
other: Not classified according to CLP Regulation (EC 1272/2008)
Conclusions:
The dermal LD50 value of the substance in rats of both sexes was estimated to exceed 2000 mg/kg bw after 24 hours of application and over a period of 14 days.
Executive summary:

The dermal acute toxicity of the substance was evaluated with a limit test to albino rat Wistar, according to the OECD Guideline 402 (1987). A 2000 mg/kg bw single dose of the substance was suspended in an aqueous solution and was administered to two groups each of 5 males and 5 females. A gauze patch fixed to aluminium foil and flexible bandage with petrolatum was applied for 24 h to the skin previously shaved. Symptoms, skin irritations and mortality after administration were recorded during the observation period of 14 days. Thereafter all animals were submitted to necropsy and macroscopic examination. No mortality occurred during the study period.

The dermal LD50 value of the substance in rats of both sexes over a period of 14 days was estimated to exceed 2000 mg/kg bw.