Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
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other: GLP-compliant guideline study available as unpublished report, no restrictions, fully adequate for assessment.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 423 (Acute Oral toxicity - Acute Toxic Class Method)
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. certificate)
Remarks:
Toxi-Coop Zrt, Pálya u. 2., 2120 Dunakeszi, Hungary
Test type:
acute toxic class 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:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: TOXI COOP ZRT. Cserkesz u. 90. 1103 Budapest, Hungary
- Age at study initiation: 11 weeks
- Weight at study initiation: 241-256g (step 1), 240-256g (step 2)
- Fasting period before study: the day before treatment the animals were fasted. The food but not water was withheld overnight
- Housing: In groups of 3 animals/cage) in Type II polypropylene/polycarbonate cages with laboratory bedding.
- 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: 28 days (step 1) and 29 days (step 2)

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

Route of administration:
oral: gavage
Vehicle:
other: sunflower oil
Details on oral exposure:
- Justification of the doses: Starting dose was selected on the basis of the available information about the test item. The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. One female animal died and the test was continued at 2000 mg/kg bw dose level on further three female rats. No animal died in the second step, so the test was finished.
- Formulation: All doses were formulated in the vehicle. Concentration of formulations was adjusted to maintain a treatment volume of 10 mL/kg bw.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6
Control animals:
no
Details on study design:
PROCEDURE
- A single oral administration followed by a fourteen-day observation period was performed by gavage. The day before treatment the animals were fasted. The food but not water was withheld overnight. Animals were weighed before the application and the food was given back 3 hours after the treatment.
OBSERVATIONS AND EXAMINATIONS:
- Mortality: Animals were observed individually after dosing at least once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h after the treatment and twice each day for 14 days thereafter.
- General state, external appearance, behavior and clinical symptoms: Individual observations were performed on 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 observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Body weight: The body weights were recorded on day 0 (just before the treatment), on day 7 and on day 15 with a precision of 1 g.
- Necropsy: At the end of the observation period rats 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:
The method used was not intended to allow the calculation of a precise LD50 value.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
In step 1, one rat died on Day 1. The death seemed to be a consequence of the systemic toxic effect of the test item. All female rats in step 2 survived until the end of the 14-day observation period.
Clinical signs:
In group 1 clinical signs comprised of decreased activity (9 cases of 43 observations), diarrhea (14/43), piloerection (12/43) and irritability (1/43). Decreased activity (score -1; -2), diarrhea (score +1) and piloerection (score +1; +2) occurred in all animals. Irritability (score +1) was observed in one animal. The clinical symptoms were detected between the treatment day and Day 5.
In group 2 clinical signs comprised of decreased activity (9 cases of 57 observations), pain reaction (4/57), diarrhea (18/57), piloerection (17/57) and irritability (5/57). Decreased activity (score -1; -2), diarrhea (score +1) and piloerection (score +1; +2) occurred in all animals. However, pain reaction (score +1) and irritability (score +1) was observed in one animal. The clinical symptoms were detected between the treatment day and Day 4.
Body weight:
In group 1, the mean body weight of the survivor animals corresponded to their species and age throughout the study.
In group 2, one animal lost body weight between treatment day and Day 7. Although the body weight loss was very slight (approx. 2.3 %), this animal did not regain the original body weight. This animal lost 4 g body weight between Day 0 and Day 15 (approx. 1.6 %). The mean body weight of the other animals corresponded to their species and age throughout the study.
Gross pathology:
One rat spontaneously died during the study. In this animal autolysis was observed . This is considered to be a normal physiological process after death. The five other animals survived until the scheduled necropsy on Day 15. Slight hydrometra was observed in one female of the group 2. This is considered to be a physiological finding and connected to the cycle of the animal. No pathological changes were found related to the effect of the test item during the macroscopic examination of animals.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The oral LD50 of the test item, observed in a GLP compliant OECD 423 study, is >2000 mg/kg bw .
Executive summary:

In a GLP compliant acute oral toxicity study performed according to OECD 423 using the acute toxic class method, female Wistar rats were exposed in two groups of each three rats by gavage to a dose of 2000 mg/kg bw test item dissolved in sunflower oil. One animal died in group 1. Clinical signs in group 1 comprised of decreased activity, diarrhea, piloerection observed in all animals and irritability was observed in one animal. The clinical symptoms were detected between treatment day and Day 5. In group 2 decreased activity, diarrhea and piloerection occurred in all animals, but pain reaction and irritability was observed in one animal. The clinical symptoms were detected between treatment day and Day 4. One animal lost body weight in the first week, but the mean body weight of the other animals corresponded to their species and age throughout the study. Autopsy revealed no treatment related pathological changes. The LD50 of the test item was therefore determined to be > 2000 mg/kg bw.