Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
25 February 2016 to 10 March 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

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
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Test material form:
other: yellow semi-solid
Specific details on test material used for the study:
- Physical state: Yellow semi-solid
- Analytical purity:100%
- Expiration date of the lot/batch: 23 November 2017
- Storage condition of test material: room temperature in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
ANIMALS and ANIMAL HUSBANDRY
- Female Wistar rats were supplied by Envigo RMS UK Ltd, Oxon, UK.
- The animals were randomly allocated to cages on receipt.
- Females were nulliparous and non-pregnant.
- After an acclimatisation period of at least 5 days the animals were selected at random and given a unique number within the study by indelible ink-marking on the tail and the number was written on a cage card.
- At the start of the study the animals were 8 to 12 weeks of age.

- Body weight did not exceed ± 20 % of the mean body weight for each sex.
- Animals were housed individually during 24-hour exposure period and in groups of five, by sex, for the remainder of the study in suspended solid-floor polypropylene cages furnished with woodflakes.
- Free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo Research UK Ltd, Oxon, UK) was allowed throughout the study.
- Diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect purpose or integrity of the study.
- Temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70 % respectively.
- Rate of air exchange was at least 15 changes per hour.
- Lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
- Animals were provided with environmental enrichment items considered not to contain any contaminant at a level that might affect the purpose or integrity of the study.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
PROCEDURE
- On the day before treatment, the back and flanks of each animal were clipped free of hair.
- Using available information on the toxicity of the test item, a group of five male and five female rats was treated with the test item at a dose level of 2000 mg/kg
- The appropriate amount of test item was applied as evenly as possible to an area of shorn skin (approximately 10 % of the total body surface area) using a graduated syringe.
- A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage.
- The animals were caged individually for the 24-hour exposure period.
- Shortly after dosing the dressings were examined to ensure they were securely in place.
- After the 24 hour contact period the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with arachis oil BP to remove any residual test item.
- The animals were returned to group housing for the remainder of the test period.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 males and 5 females
Control animals:
no
Details on study design:
TEST ITEM PREPARATION
- The test item was weighed out according to each animal’s individual body weight.
- Absorption of the test item was not determined.


Statistics:
- Data evaluations included the relationship, if any, between exposure of the animal to the test item and the incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, body weight changes, mortality and other toxicological effects.
- Using the mortality data obtained, and estimate of the acute dermal median lethal dose (LD50) of the test item was made.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- There were no deaths
Clinical signs:
- No signs of systemic toxicity were noted during the observation period (see Appendix 1, attached).
Body weight:
- Individual body weights and body weight changes are shown in Appendix 4 (attached).
- All animals showed expected gains in body weight over the observation period.
Gross pathology:
- Individual necropsy findings are given in Appendix 5 (attached).
- No abnormalities were noted at necropsy.
Other findings:
DERMAL REACTIONS
- Individual dermal reactions are shown in Appendix 2 and 3 (attached).
- Very slight to well-defined erythema and very slight edema were noted at the test sites of one male and all females.
- Very slight erythema, with or without very slight edema was noted at the test sites of four males.
- Crust formation was noted at the test sites of all animals.
- Small superficial scattered scabs were noted at the test sites of two females with desquamation also noted at the test sites of two females.
- Treated skin sites of males appeared normal 7 days after dosing and the treated skin sites of four females appeared normal 7 or 14 days after treatment.
- Small superficial scattered scabs were still present at the treatment site of one female at the end of the observation period on Day 14.

Applicant's summary and conclusion

Interpretation of results:
other: not classified
Remarks:
EU
Conclusions:
The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be greater than 2000 mg/kg body weight.
Executive summary:

INTRODUCTION

The study was performed to assess the acute dermal toxicity of the test item in the Wistar strain rat.

METHODS

A group of then animals (five males and five females) was given a single, 24-hour, semi-occluded dermal application of the test item to intact skin at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

RESULTS

Mortality: There were no deaths.

Clinical observations: There were no signs of systemic toxicity.

Dermal irritation: Signs of dermal irritation noted were very slight to well-defined erythema, very slight edema, crust formation, desquamation and small superficial scattered scabs. Treated skin sites of males appeared normal 7 days after dosing and the treated skin sites of four females appeared normal 7 or 14 days after treatment. Small superficial scattered scabs were still present at the treatment site of one female at the end of the observation period on Day 14.

Body weight: All animals showed expected gains in body weight.

Necropsy: No abnormalities were noted a necropsy.

CONCLUSION

The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be greater than 2000 mg/kg body weight.