Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
January 20 - February 17, 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): E-BW102
- Physical state: Red brown colored powder
- Storage condition of test material: Room temperature in an air-tight container in a dark place
- Stability under storage conditions: No data

Test animals

Species:
rat
Strain:
other: Crl:CD
Sex:
male/female
Details on test animals and environmental conditions:
- Source: Charles River Laboratories Japan, Inc. (Hino Breeding Center)
- Age at study initiation: 7 week old males and 10 week old females
- Weight at study initiation: 223.6 to 236.9 (males), 217.6 to 250.8 (females)
- Housing: Individually housed in labeled stainless-steel cages (WxDxH: 226x346x198 mm)
- Diet (e.g. ad libitum): Free access to autoclave-sterilized pellet diet (CRF-1, Oriental Yeast Co., Ltd.)
- Water (e.g. ad libitum): Free access to well water admixed with NaClO (about 2 ppm).
- Acclimation period: The acclimatization period was set for 8 days.
- Health inspection: During the acclimatization period, all animals were examined for clinical signs (once daily) and body weights (on the day of receipt and 5 days after receipt) to confirm their health status.

Results of analysis for diet (lot no. 101005) was provided by Oriental Yeast Co., Ltd., and the contaminants in the diet were confirmed to be within the acceptable limits established by the test facility. The water was analyzed twice a year at Nichigo Kyushu Co., Ltd. The results were confirmed to be in compliance with the SOP of the test facility.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.9 – 23.8
- Humidity (%): 48.4 - 57.0
All housing equipment was sterilized by autoclaving after washing water. The floor of the animal room was cleaned and wiped every day with a disinfectant-soaked mop. NaClO was used as disinfectant.
- Air changes (per hr): 10 to 20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: February 03, 2011 to February 17, 2011

Administration / exposure

Type of coverage:
occlusive
Vehicle:
other: moistened with an appropriate amount (1.0 mL) of water for injection
Details on dermal exposure:
The fur on the dorsal skin (areas of about 5 x 6 cm) was clipped off with electric clippers, taking care not to injure the skin, at about 24 hours before dosing.

The animals were fitted with a neck collars after the hair clipping. The test substance application area was prepared to be within about 10% of the total body surface (about 4 x 5 cm). The required amount of the test substance was applied uniformly on a lint cloth (about 4 x 5 cm) lined with an impermeable sheet. The lint cloth was moistened with an appropriate amount (1.0 mL) of water for injection. The lint cloth loaded with the test substance was applied to the skin, and the application area was covered with an elastic adhesive bandage for about 24 hours. After about 24 hour exposure, the test substance, lint cloth and adhesive bandage were removed.

Frequency: Single dosage, on Day 1.

Washing: The application area was wiped with a gauze sheet immersed in lukewarm water and water was removed with dry tissue paper.

The animals were fitted with the neck collars during the dosing and recovery periods.
Duration of exposure:
24 hours.
Doses:
2000 mg/kg


No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
DOSAGE PREPARATION: The prescribed amount of the test substance was calculated based on the body weights on the day of dosing.

Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Body weights: All animals were subjected to the body weight measurement before dosing (day 1) and on days 4, 8 and 15. Moreover the body weight gain between each measurement day was calculated.
Mortality/Clinical signs: All animals were observed for their mortality and clinical signs before the dosing and at 30 minutes, 1, 3 and 6 hours postdose on the day of the dosing and once daily for 14 days thereafter.
- Necropsy of survivors performed: At the end of the 14-day observation period, all animals were euthanized by exsanguination under intravenous sodium pentobarbital (30 mg/kg) anesthesia and dissected immediately and all organs and tissues were examined macroscopically.
Statistics:
None.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths occurred.
Clinical signs:
No abnormalities including skin irritation was seen in either sex.
Body weight:
Although decreased body weight, which was considered to be attributable to the occlusive application or the fitted neck collar, was seen in 1 female on day 4, body weights of both sexes increased steadily on and after day 8.
Gross pathology:
No abnormalities were seen in either sex.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an acute dermal toxicity study performed according to OECD 402 and GLP, the LD50 value of E-BW102 in male and female Crl:CD(SD) rats was established to exceed 2000 mg/kg body weight.
According to Regulation (EC) No. 1272/2008, E-BW102 needs not to be classified for acute dermal toxicity.