Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From January 12, 2009 to January 26, 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2009

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
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Test animals

Species:
rat
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
Number of Animals: 10
Sex: 5 Males and 5 Females. Females assigned to test were nulliparous and nonspregnant.
Species/Strain: Rats/Sprague-Dawley derived, albino.
Age/Body weight: Young adult (10-11 weeks)/males 359-382 grams and females 222-240 grams at experimental start.
Source: Received from Ace Animals, Inc., Boyertown, PA on December 18, 2008.
Housing: The animals were singly housed in suspended stainless steel caging with mesh floors, which conform to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals DHEW (NIH). Litter paper was placed beneath the cage and was changed at least three times per week.
Animal Room Temperature and Relative Humidity Ranges: 18-22ºC and 12-27%, respectively. The low humidity occurred during the study due to a temporary malfunction of the environmental control system.
Photoperiod: 12-hour light/dark cycle
Acclimation Period: 25 days
Food: Purina Rodent Chow #5012
Water: Filtered tap water was supplied ad-libitum by an automatic water dispensing system.
Contaminants: There were no known contaminants reasonably expected to be found in the food or water at levels, which would have interfered with the results of this study. Analyses of the food and water are conducted regularly and the records are kept in the laboratory facility.
Cage: Each cage was identified with a cage card indicating at least the study number and identification and sex of the animal.
Animal: A number was allocated to each rat on receipt and a stainless steel ear tag bearing this number was attached to the animal. This number, together with a sequential animal number assigned to study 26612, constituted unique identification.

Administration / exposure

Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Preparation and Selection of Animals
On the day prior to application, a group of animals was prepared by clipping the dorsal area and the trunk. After clipping and prior to application, the animals were examined for health, weighed (initial), and the skin checked for any abnormalities. Ten healthy, naive rats (five males and five females; not previously tested) were selected for test.

Dose Calculations
Individual doses were calculated based on the initial body weights, taking into account the specific gravity (determined by EPSL) of the test substance.

Application of Test Substance
Five thousand mg/kg of body weight of the test substance was applied evenly over a dose area of approximately 2 inches x 3 inches (approximately 10% of the body surface) and covered with a 2-inch x 3-inch, 4-ply gauze pad. The gauze pad and entire trunk of each animal were then wrapped with 3-inch Durapore tape to avoid dislocation of the pad and to minimize loss of the test substance. The rats were then returned to their designated cages. The day of application was considered Day 0 of the study. After 24 hours of exposure to the test substance, the pads were removed and the test sites were gently cleansed of any residual test substance.
Duration of exposure:
24 hours.
Doses:
5000 mg/kg bw
No. of animals per sex per dose:
10
Control animals:
not required
Details on study design:
Body Weights
Individual body weights of the animals were recorded prior to test substance application (initial) and again on Days 7 and 14 (termination).

Cage-Side Observations
The animals were observed for mortality, signs of gross toxicity, and behavioral changes during the first several hours after application, and at least once daily thereafter for 14 days.
Observations included gross evaluation of skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behavior pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhea, and coma.

Necropsy
All rats were euthanized via CO2 inhalation on Day 14. Gross necropsies were performed on all animals. Tissues and organs of the thoracic and abdominal cavities were examined.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortalities were observed.
Clinical signs:
Following application, all animals exhibited facial staining and/or ano-genital staining between Days 1 and 2. Also, dermal irritation (erythema, edema, desquamation, hyperkeratosis, discoloration, eschar and/or corrosion) was noted at the dose site of all animals between Days 1 and 14.
Body weight:
All animals survived exposure to the test substance and gained body weight during the study.
Gross pathology:
Gross necropsy of the females revealed red intestines. No gross abnormalities were noted for any of the males when necropsied at the conclusion of the 14-day observation period.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the study conditions, the acute dermal LD50 of the test substance was greater than 5000 mg/kg bw in male and female rats.
Executive summary:

A study was conducted to determine the potential acute toxicity of the test substance by the dermal route according to OECD Guideline 402, EU Method B.3 and EPA OPPTS Method 870.1200, in compliance with GLP. the test susbtance was applied a single time at 5000 mg/ kg bw to the skin of 10 healthy rats for 24 h. The animals were observed for mortality, signs of gross toxicity and behavioral changes at least once daily for 14 d. Body weights were recorded prior to application and again on Days 7 and 14 (termination). Necropsies were performed on all animals at terminal sacrifice. All animals survived exposure to the test substance and gained body weight during the study. Following application, all animals exhibited facial staining and/or ano-genital staining between Days 1 and 2. Also, dermal irritation (erythema, edema, desquamation, hyperkeratosis, discoloration, eschar and/or corrosion) was noted at the dose site of all animals between Days 1 and 14. Gross necropsy of the females revealed red intestines. No gross abnormalities were noted for any of the males when necropsied at the conclusion of the 14 d observation period. Under the study conditions, the acute dermal LD50 of the test substance was greater than 5000 mg/kg bw in male and female rats (Oley, 2010).