Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April 26, 1994 - May 10, 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
1987
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
1992
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wñlferstrasse 4, CH-4414 Füllinsdorf/Switzerland
- Age at study initiation: males at 9 weeks; females at 11 weeks
- Weight at study initiation: males at 256.0 - 269.6; females at 189.0 - 209.7 g
- Diet: pelleted standard Kliba 343. Batch nos. 86/93 and 87/93 rat maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst) available ad libitum (except for over-night fasting period prior to application).
- Water: community tap water from Fülllnsdorf, available ad libitum.
- Acclimation period: one week under laboratory conditions» after health examination. Only animals without any visual signs of illness were used for the study.
- Identification: by unique cage number and corresponding color-coded spots on the tail.
- Randomization: randomly selected at time of delivery Into groups of five

ENVIRONMENTAL CONDITIONS
- Temperature: continuously monitored environment with a temperature of 22 ± 2 °C
- Humidity: 36 - 64 %
- Air changes: 10-15 ACH
- Photoperiod: 12 hours artificial fluorescent light (approx. 100 Lux) / 12 hours dark (light period between 6.00 a.m. to 6.00 p.m.), music during the light period
- Cage: during acclimatization in groups of five in Makrolon type-4 cages (size: 33 x 55 x 20 cm) with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz); during treatment and observation individually in Makrolon type-3 cages (size: 22 X 37.5 x 15 cm) with standard softwood bedding.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
bi-distilled
Details on dermal exposure:
TEST SITE
- Area of exposure: approximately 24 hours before treatment, the backs of the animals were clipped with an electric clipper. On test day 1, the test article was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe.
- % coverage: area of approximately 10 % of the total body surface.
- Type of wrap if used: semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.
- Rationale: dermal administration was used because this is one possible route of human exposure during manufacture, handling and use of the test article.

REMOVAL OF TEST SUBSTANCE
- Washing: the skin was washed with lukewarm tap water and dried with disposable paper towels.
- Time after start of exposure: twenty-four hours after the application.

TEST MATERIAL
- Amount applied: 4.0 ml/kg
- For solids, paste formed: no
- Preparation: the test article was placed into a glass beaker on a tared Mettier PM 480 balance, and the vehicle, (bi-distilled water) was added. A weight by volume dilution was prepared using a homogenizer (Ultra-Turrax, Janke & Kunkel, D-79219 Staufen).
Homogeneity of the test article in the vehicle was maintained during treatment using a magnetic stirrer (Janke & Kunkel, D-79219 Staufen).
The preparation was made shortly before dosing.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw, single dose
No. of animals per sex per dose:
5 males and 5 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing: on test day 1 (pre-administration), 8 and 15.
- Necropsy: necropsies were performed by experienced prosectors. At the end of the observation period all animals were anesthetized by intraperitoneal injection of NARCOREN at a dose of at least 2.0 ml/kg body weight and sacrificed by exsanguination. The animals were examined macroscopically.
- Other examinations performed: each animal was examined for changes in behaviour and appearance (with special emphasis on the application area, except for the time when the semi-occlusive dressing was in place) four times during day 1, and once dally during days 2-15.
Statistics:
The LOGIT-Model could not be used as no deaths occurred,

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: Value not quantified since higher than tested concentrations
Mortality:
No deaths observed.
Clinical signs:
No clinical signs of systemic toxicity.
Body weight:
Not affected by tratment with test substance. The slight reduction in body weight gain during the first observation week was considered to be a consequence of the semi occlusive dressing.
Gross pathology:
No organ abnormalities were observed at necropsy.
Other findings:
A red discoloration of the skin at the application site was evident in all animals after the removal of the dressing on test day 2 and persisted until test day 3 - 6.

Applicant's summary and conclusion

Interpretation of results:
other: not classified according to the CLP Regulation (EC 1272/2008)
Conclusions:
LD50 (males and females) > 2000 mg/kg b.w.
Executive summary:

The test article was administered to a group of 5 male and 5 female rats by semi-occlusive dermal application with a single dose of 2000 mg test article/kg body weight.

- No deaths occurred during the study period.

- No clinical signs of systemic toxicity were observed during the observation period.

- A red discoloration of the skin at the application site was evident in all animals after the removal of the dressing on test day 2 and persisted until test day 3 - 6.

- There were no test article-related effects on the body weight of the animals during the observation period. The slight reduction in body weight gain during the first observation week was considered to be a consequence of the semi occlusive dressing.

- The macroscopic examination at study termination revealed no organ abnormalities.

In conclusion, the mean lethal dose after single oral administration to rats of both sexes, observed over a period of 14 days, could not be estimated, because no deaths occurred at the maximal dose of 2000 mg test article/kg body weight.