Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
8 January to 28 January 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted in accordance with OECD, EU and US guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
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
Remarks:
migrated information: powder
Details on test material:
Chemical Name: Zinc 3,5-bis(α-methylbenzyl)salicylate
CAS number: 53770-52-8

Test animals

Species:
rat
Strain:
other: CRL:(WI)
Sex:
male/female
Details on test animals and environmental conditions:
Species and strain: CRL:(WI) rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Justification of strain: The Wistar rat is one of the standard rodent species used in acute toxicity studies
Number of animals: 5 animals/sex
Sex: Male and female, female rats were nulliparous and non-pregnant.
Age of animals at study start: Young adult rats
Body weight range at dosing: Between 219 g and 236 g
Acclimatization time: 6 days

Food and Water Supply
Animals received ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany, ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottle, ad libitum. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study and the water was considered fit for human consumption.
The batch of feed employed in the study was as follows: 680 2237, expiry date: March 2015
The supplier provided an analytical certificate for the batch used. Copy of the certificates will be archived with the raw data.
Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

Animal Identification
The individual identification was performed using numbers written on the tail with a marker pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.' s Master File for each animal allocated to the treatment groups. The cages were identified by cards containing information about study code, sex, dose group, cage number and individual animal number.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
A single dermal application was made and was followed by a fourteen-day observation period. Sufficient water was used to dampen the test material to ensure good contact with the skin.

Procedure
The back of each animal was shaved (approximately 10 % area of the total body surface) approximately 24 hours prior to treatment. The test item was applied as a single dose, moistened with water to the shaved skin and remained in contact with the skin for the 24-hour exposure period. Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin using a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.
At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
no
Details on study design:
Clinical Observations
Clinical observations were performed on the day of treatment at 1 and 5 hours after application of the test item and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Skin Irritation
Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.

Measurement of Body Weight
The body weights were recorded on Day 0 (before test item administration) and on Days 7 and 14 just before necropsy.

NECROPSY
Macroscopic examination was performed on all animals. All animals were anaesthetised with pentobarbital sodium (details in 3. 3.) and exsanguinated. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.

Results and discussion

Preliminary study:
Not applicable
Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Test item did not cause mortality at the dose level of 2000 mg/kg bw.
Clinical signs:
There were no systemic clinical signs noted in any animal throughout the study.
Body weight:
There were no treatment related effects on body weight or body weight gain during the observation period. However a slight body weight loss was noted in one female rat at the dose level of 2000 mg/kg bw between Days 7 and 14.
Gross pathology:
No macroscopic observations were noted at a dose level of 2000 mg/kg bw.
Other findings:
No local dermal signs were observed after treatment with the test item during the 14 days observation period.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute dermal median lethal dose (LD50) of the test item was found to be greater than 2000 mg/kg body weight in male and female CRL:(WI) rats.
According to the CLP Regulation, the test item is not classified for acute dermal toxicity.
Executive summary:

An acute dermal toxicity study was performed with test item in CRL:(WI) rats, in compliance with OECD Guideline No.: 402.

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied as a single dermal 24-hour exposure followed by a 14-day observation period. Sufficient water was used to dampen the test material to ensure good contact with the skin.

Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Gross macroscopic examination was performed on all animals at the end of the 2-week observation period (Day 14).

The results of the study were summarized as follows:

Mortality

Test item did not cause mortality at the dose level of 2000 mg/kg bw.

Systemic clinical signs

There were no systemic clinical signs noted in any animal throughout the study.

Local dermal signs

No local dermal signs were observed after treatment with the test item during the 14 days observation period.

Body weight and body weight gain

There were no treatment related effects on body weight or body weight gain during the observation period. However, a slight body weight loss was noted in one female rat at the dose level of 2000 mg/kg bw between Days 7 and 14.

Necropsy

No macroscopic observations were noted at a dose level of 2000 mg/kg bw.

The acute dermal median lethal dose (LD50) of the test item was found to be greater than 2000 mg/kg body weight in male and female CRL:(WI) rats.

According to the CLP Regulation, the test item is not classified for acute dermal toxicity.