Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April to May 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
not specified
GLP compliance:
yes
Test type:
acute toxic class method

Test material

Reference
Name:
Unnamed
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
cool, good aired

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Dobrá Voda, Slavak Republik
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9-11 weeks
- Housing: the animals were housed in plastic cages suspended on stainless steel racks, 3 animals per cage in a rrom equipped with central air-conditioning.
- Diet (e.g. ad libitum): The laboratory food ssniff (Spezialdiäten GmbH, Germany) was offered at recommended doses each day approximately at the same time.
- Water (e.g. ad libitum): The animals received tap water for human consumption. Supply of drinking was unlimited. The quality of drinking water is periodically analysed and recorded.
- Acclimation period: The animals were acclimated under the conditions identical to the conditions during the experiment 5 days prior to the start of treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.86 +/- 0.54 °C
- Humidity (%): 54.45 +/- 3.07 %
- Air changes (per hr): not mentioned
- Photoperiod (hrs dark / hrs light): 12h / 12h

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 5ml/kg bw

Doses:
2.000 mg/kg (3 animals)
300 mg/kg (6 animals)
50 mg/kg (6 animals)
No. of animals per sex per dose:
3-6
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed individually immediately after administration of the test item and 0.5, 1, 2, and 4 hours later. Each animal was inspected daily for the next 14 days. Observations included: changes in skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity, and behavioural pattern. Particular attention was given to potential neurologic endpoints such as tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Necropsy of survivors performed: yes

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 50 - <= 300 mg/kg bw
Mortality:
2000 mg/kg bw: 3/3 animals died
300 mg/kg bw: 3/6 animals died
50 mg/kg bw: 0/6 animals died
Clinical signs:
2000 mg/kg bw: All animals died within 10 minutes after application.
300 mg/kg bw: piloerection and lethargy were noted in several animals.
50 mg/kg bw: piloereaction and lethargy was noted in one animal.
Body weight:
The body weights of all survived animals increased during the study. A slight decrease of body weight in two animals and a stagnation of body weight in one animal of the low dose group were observed between the first and second week after administration of the test item.
Gross pathology:
All animals were necropsied. Gastric erosions and hyperemia of gastrointestinal tract were registered in animals treated with the test item at the dose of 2000 mg/kg body weight and in dead animals treated with the dose of 300mg/ kg body weight. In addition, splenomegaly was observed in one animal of the high dose group. One animal of the mid dose group could not be necropsied because cadaver was autolysed. During necropsy, no macroscopic findings were noticed in animals treated with the dose of 50 mg/kg body weight.

Applicant's summary and conclusion

Interpretation of results:
Category 3 based on GHS criteria
Conclusions:
The LD50 of the test item is higher than 50 mg/kg bw and lower than 300 mg/kg bw after single oral administration to Wistar rats.
Executive summary:

The purpose of the study was to evaluate the potential toxic effect of the test item when administered as a single oral dose to Wistar rats. The procedure according to OECD Guideline 423 Acute Toxic Class (ATC) Method was used. A limit dose of 2000 mg/kg body weight was used as a starting dose. One group of 3 females was dosed. Test item-related mortality was observed within few minutes. In

a second step, 3 females were treated at the dose of 300 mg/kg body weight. Only one animal died within five days and therefore another 3 females (third step) were treated at the same dose of 300

mg/kg body weight. Test item-related mortality was observed within 2 hours after administration of the test item. During this observation period, two animals died. In a fourth step, 3 females were treated

at the dose of 50 mg/kg body weight. All females survived 24 hours and therefore another 3 females (fifth step) were treated at the same dose of 50 mg/kg body weight. The test item administered to 3 females at a limit dose of 2000 mg/kg body caused death of 3/3 animals. Gastric erosions, hyperemia of gastrointestinal tract and splenomegaly were observed. 3/6 females survived the dose of 300 mg/kg. Lethargy and piloerection were observed during the observation period. A stagnation of body weight in one animal was observed between the first and second week after administration of the test item. During necropsy of the dead animals, the same findings as in animals which were treated with the dose of 2000 mg/kg body weight were registered. Piloerection and lethargy were noted in one animal treated with the dose of 50 mg/kg body weight, the rest animals did not display signs of toxicity during the first 4 hours and the 14-day observation period. The body weight of all survived animals increased during the study. A slight decrease of body weight in two animals and a stagnation of body weight in one animal treated with the dose of 50 mg/kg body weight were observed between the first and second week after administration of the test

item. During necropsy, no macroscopic findings were observed. The LD50 of the test item after single oral administration to Wistar rats is higher than 50 mg/kg body weight and lower than 300 mg/kg body weight.