Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-compliant guideline study available as unpublished report, no restrictions, fully adequate for assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
Remarks:
Toxi-Coop Zrt, Pálya u. 2, 2120 Dunakeszi, Hungary
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Petol PM 410-4N
- Physical state: yellow-brown (apparently yellow), viscous, clear liquid
- Storage condition of test material: Store in tightly closed container, in a dry and well ventilated area, between 20-30°C.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90.
- Age at study initiation: Young adult rats, females were nulliparous and non-pregnant
- Weight at study initiation: Preliminary study: 259 - 295 g; Main test: male : 254 - 268 g; female: 239 - 296 g
- Housing: During acclimatization: 3 animals/sex/cage. During the study: individually in type II polypropylene/polycarbonate rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets.
- Diet: Ssniff® SM R/M-Z+H complete diet for rats and mice produced by ssniff Spezialdiäten GmbH, D-59494, Soest, Germany, ad libitum
- Water: tap water from municipal supply, ad libitum
- Acclimation period: Preliminary study: 54 days in preliminary study; Main study: Females: 68 days, Males: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 8-12 air exchanges/hour by central air-condition system.
- Photoperiod (hrs dark / hrs light): Artificial light, from 6 a.m. to 6 p.m.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
other: sunflower oil
Details on dermal exposure:
The backs of the animals were shaven (approximately 10 % area of the total body surface) 24 hours prior to the treatment. The test item was applied in a single 2000 mg/kg bw dose uniformly over the shaved skin throughout a 24- hour exposure period. Sterile gauze pads were placed on the skin of the rats. These gauzes were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was wrapped with semi-occlusive plastic wrap for 24 hours. At the end of the exposure period, residual test item was removed, using body temperature water.
Duration of exposure:
24 hours
Doses:
Preliminary test: 5, 50, 300, 2000 mg/kg bw
Main test: 2000 mg/kg bw
No. of animals per sex per dose:
Preliminary test: 2 (females)
Main test: 5
Control animals:
no
Details on study design:
Observations:
- Mortality: Inspection for signs of morbidity and mortality were made twice daily at the beginning and end of the working day.
- Clinical observations: Careful clinical observation was made at the following intervals: 1h, 5h after the treatment and once each day for 14 days thereafter. Individual observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Body weight: The body weight was recorded on day 0 (shortly before treatment), on day 7 and on day 14 with a precision of 1 g in the main study.
- Pathology: At the end of the observation period all rats of the main study were sacrificed under isofluran anaesthesia. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed, and any abnormality was recorded with details of its location, colour, shape and size.
Statistics:
No statistical analysis has been performed.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
No behavioural changes or systemic toxic signs were noted during the study. Dermal irritation symptoms as erythema and other signs were observed in all animals on the treatment site. The redness was detected between Day 1 and Day 7 in males and between Day 1 and Day 14 in females. The redness was very slight to well defined (score 1 and 2) in males and very slight to severe (score 1 to 3) in females. Other dermal irritation signs were as follows: wounds and crusting. Wounds were recorded between Day 2 and Day 7 in males and between Day 1 and Day 14 in females. Wounds appeared in all animals. Crusting was detected in males between Day 7 and Day 12.
Body weight:
The mean body weight of the male animals corresponded to their species and age throughout the study. A body weight loss was observed in all females between treatment day and Day 7 and in one female between Day 7 and Day 15. Although these body weight losses were below 10%, the possibility that this effect is a result of the toxic effect of the test item could not be excluded, because four animals did not regain the original body weight.
Gross pathology:
All animals survived until the scheduled necropsy on Day 14. No pathological changes were found related to the effect of the test item during the macroscopic examination of male animals. No macroscopic alterations due to the systemic toxic effects of the test item were found in females, but external necropsy findings were detected on site of the administration. Wounds on the treated area were observed in all females. Very slight redness were recorded in two animals, well defined erythema was detected in one animal and moderate to severe erythema was observed in two animals. Slight hydrometra was observed in two females and moderate hydrometra was found in two females. Hydrometra was an indication for the sexual cycle of female animals and is a frequent observation in experimental rats with no toxicological meaning.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The dermal LD50 of the test item, observed in a GLP compliant OECD 402 study, is >2000 mg/kg bw .
Executive summary:

In a GLP compliant acute dermal toxicity study performed according to OECD 402, 5 male and 5 female Wistar rats were exposed to the test item at 2000 mg/kg bw by the dermal route. The test item, dissolved in sunflower oil was applied semi-occlusive and left in contact with the skin for 24 hours, followed by a 14-day observation period. No mortalities occurred. No behavioural changes or systemic toxic signs were noted during the study. Dermal irritation symptoms as very slight to well defined erythema in males and very slight to severe erythema in females were observed on the treatment site. These symptoms were observed in the first week in males and throughout the 14-day observation period in females. Other signs as wounds and/or crusting appeared in males between Day 2 and Day 12. Wounds occurred in all females between Day 1 and Day 14. The body weight development was undisturbed in all male animals. The body weight loss was observed in all females in the first week and in one female in the second week. Four animals did not regain its original body weight. No treatment-related macroscopic alterations of organs and tissues were seen during necropsy. The external alterations (erythema, wounds) occurred in females were in line with the observed local irritant symptoms. Under the experimental conditions, the acute dermal LD50 value of

the test item is determined to be greater than 2000 mg/kg bw.