Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From September 27 to October 18, 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report Date:
1990

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
adopted February 24th, 1987
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd.
- Age at study initiation: 10 weeks males; 12 weeks females.
- Weight at study initiation: 211 - 243 g males; 189 - 205 g females.
- Housing: individually in Makrolon type-2 cages with standard softwood bedding.
- Diet: pelleted standard Kliba 343, rat maintenance diet, ad libitum.
- Water: community tap water from Itingen, ad libitum.
- Acclimation: one week under laboratory conditions and after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 40 - 70 %
- Air changes: 10-15 air changes per hour.
- Photoperiod: 12 hrs dark / 12 hrs artificial fluorescent light.
- Other: music during the light period, room was air-conditioned.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Preparation of site of exposure: approximately 24 hours before treatment, the backs of the animals were clipped with an electric clipper.
- % coverage: appr. 10 % of the total body surface.
- Application of test material: applied evenly on the skin with a syringe.
- Type of wrap if used: semi - occlusive dressing wrapped around the abdomen and fixed with an elastic adhesive bandage.

TEST MATERIAL
- Amount volume: 4 ml at 2000 mg/kg
- Preparation of test sample: the test article was placed into a glass beaker on a tared balance and the vehicle (bi-distilled water) was added. A weight/volume dilution was prepared using a homogenizer. Homogeneity of the test article in the vehicle was maintained during treatment using a magnetic stirrer. The preparation was made immediately prior to dosing.

REMOVAL OF TEST SUBSTANCE
- Washing: with lukewarm tap water. The skin was then dried with disposable paper towels.
- Time after start of exposure: 24 hours after the application.
Duration of exposure:
24 hours.
Doses:
2000 mg/kg bw.
No. of animals per sex per dose:
5 male and 5 females.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 15 days.
- Frequency of observations: mortality and viability were recorded four times during test day 1 and daily during days 2 - 1 5.
- Frequency of weighing: on the first day (pre-administration), at day 8 and day 15.
- Clinical signs observed: each animal had an examination for changes in appearance and behaviour four times during day 1 and daily during days 2-15. All abnormalities were recorded. Due to the 24 hour semi-occlusive treatment, the local findings were observed starting with day 2 of test. The animals were checked for the following clinical signs: aggressiveness, vocalization, restlessness / excitation, nervousness, fear, sedation, somnolence, sleep, coma, rhinorrhea, epistaxis, apnea, dyspnea, rales, chromodacryorrhea, exophthalmos, miosis, mydriasis, whitish discharge, lid adhesion, negative corneal reflex, akinesia, ataxia, dropped head, hyperkinesia, hypokinesia, paralysis flaccid, paralysis spastic, paddling movements, stiff gait, rolling movements, ventral body position, latero-abdominal position, hunched posture, spasms, tonic muscle spasms, clonic muscle spasms, opisthotonus, saltatory spasms, trismus, tremor, muscle-twitching generalized, erythema, edema, necrosis, crusts, scale formations, loss of weight, diarrhea, emaciation, ruffled fur, necrosis of tissue of application area, salivation, pallor, cyanosis.
- Necropsy: all animals were necropsied. The surviving animals were euthanised at the end of the observation period by intraperitoneal injection of sodium pentobarbitone.
Statistics:
The LOGIT-Model could not be applied to the observed rate of death. The toxicity was estimated without the use of a statistical model.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality observed.
Clinical signs:
The rats exhibited blackened skin throughout the entire study period. No systemic signs were observed in any animal.
Body weight:
The body weight gain of the animals was not affected throughout the study by test article treatment.
Gross pathology:
No obvious macroscopical organ findings noted.

Any other information on results incl. tables

Clinical signs

Summary

Test day

1 1 1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Time after treatment (hrs)

1 2 3 5
Skin black (back) - Males - - - - X X X X X X X X X X X X X X
Skin black (back) - Females - - - - X X X X X X X X X X X X X X

X = sign observed

- = observation performed, sign not observed

Test day

1 1 1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Time after treatment (hrs)

1 2 3 5
Anima N.  Sign (G)
1M Skin black (back) 1 - - - - 1 1 1 1 1 1 1 1 1 1 1 1 1 1
2M Skin black (back) 1 - - - - 1 1 1 1 1 1 1 - - - - - - -
3M Skin black (back) 1 - - - - 1 1 1 1 1 1 1 1 1 1 1 1 1 1
4M Skin black (back) 1 - - - - 1 1 1 1 1 1 1 - - - - - - -
5M Skin black (back) 1 - - - - 1 1 1 1 1 1 1 1 1 1 1 - - -
6F Skin black (back) 1 - - - - 1 1 1 1 1 1 1 1 1 1 1 1 1 1
7F Skin black (back) 1 - - - - 1 1 1 1 1 1 1 1 1 1 1 1 1 1
8F Skin black (back) 1 - - - - 1 1 1 1 - - - - - - - - - -
9F Skin black (back) 1 - - - - 1 1 1 1 - - - - - - - - - -
10F Skin black (back) 1 - - - - 1 1 1 1 1 1 1 - - - - - - -

- = observation performed, sign not observed

(G) = max. grade

Applicant's summary and conclusion

Interpretation of results:
other: not classified, according to the CLP Regulation (EC) No 1272/2008
Conclusions:
LD50 (males and female) > 2000 mg/kg bw
Executive summary:

The purpose of the dermal toxicity study was to assess the toxicological profile of test item when administered to rats by a single semi-occlusive dermal application, with an observation period of 15 days. The test was perfromed accodring to the OECD guideline 402. The test article was applied to the skin of rats of both sexes for 24 hours at a single dose of 2000 mg/kg bw. The substance was removed after 24 hours exposure and the skin was washed. All the animals were subjected to necropsy at the end of the observation period.

No dead occurred during the experiment and no macroscopic findings were noted. All animals, showed a black-coloured back, from the 2nd day until the end of the observation period, but no mortality occured.

Conclusion

LD50 (males and female) > 2000 mg/kg bw