Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 September 2017 to 27 September 2017
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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Appearance: Natural white powder
- Storage conditions: Room temperature (15-25 °C), protected from light and humidity

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 11 weeks old
- Weight at study initiation: 230 - 243 g
- Fasting period before study: On the night before treatment, the animals were fasted. The food, but not water, was withheld during an overnight period. The food was returned 3 hours after the treatment.
- Housing: 3 animals/ cage in Type II polypropylene/polycarbonate cages
- Diet: Ad libitum, except for the night before treatment
- Water: Animals received tap water from the municipal supply from 500 mL bottles, ad libitum
- Acclimation period: At least 26 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2 - 25.0 °C
- Humidity (%): 30 - 80 %
- Air changes (per hr): 15 - 20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
methylcellulose
Remarks:
1 % Methyl cellulose
Details on oral exposure:
VEHICLE
- Concentration in vehicle: The test material was formulated in 1 % methyl cellulose at a concentration of 200 mg/mL

MAXIMUM DOSE VOLUME APPLIED: The dose volume was 10 mL/kg bw

DOSAGE PREPARATION: The test material was freshly formulated at a concentration of 200 mg/mL in the vehicle in the Pharmacy of Citoxlab Hungary Ltd. on the day of administration. The formulation container was stirred continuously with a magnetic stirrer until dose administration procedures were complete.

CLASS METHOD
- Rationale for the selection of the starting dose: The initial dose level was selected by the Study Director to be that which is most likely to produce mortality in some of the dosed animals. Limit dose of 2 000 mg/kg bw was selected as a starting dose. Initially, 3 female animals were treated at dose level of 2 000 mg/kg bw. No mortality was observed, therefore a further 3 animals were treated at the dose level of 2 000 mg/kg bw. As no mortality was observed in the second dose group, further testing was not required according to the test guidelines (OECD 423, Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris).
Doses:
2 000 mg/kg bw
No. of animals per sex per dose:
6 animals, 3 animals/ group
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter.
- Necropsy of survivors performed: Yes. Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthanimal 40 %). After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.
- Other examinations performed: Individual observations were performed on the skin, fur, eyes, mucous membranes, 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.
Statistics:
The method used was not intended to allow the calculation of a precise LD50 value.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed at dose level of 2 000 mg/kg bw.
Clinical signs:
All animals were symptom-free during the 14-day observation period at a dose level of 2 000 mg/kg bw.
Body weight:
There were no effects on body weights or body weight gains that could be attributed to treatment with the test material.
Gross pathology:
There was no evidence of the macroscopic observations in the animals dosed at 2 000 mg/kg bw and terminated on Day 14.

Applicant's summary and conclusion

Interpretation of results:
other: Not classified in accordance with EU criteria.
Conclusions:
Under the conditions of this study, the acute oral LD50 value of the test material was > 2 000 mg/kg bw in rats.
Executive summary:

The single-dose oral toxicity of the test material was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.Tris) in Crl:WI Wistar rats. Two groups of 3 female Crl:WI rats were treated with the test material at a dose level of 2 000 mg/kg bw (Group 1 and Group 2).

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test material was formulated in 1 % methyl cellulose at a concentration of 200 mg/mL at a dose volume of 10 mL/kg bw.

Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0, 7 and 14 (before necropsy). All animals were subjected to a necropsy and a macroscopic examination.

Initially, three females (Group 1) were treated at a dose level of 2 000 mg/kg bw. As no mortality was observed, a confirmatory group (Group 2) was treated at the same dose level. No mortality was observed in the confirmatory group, therefore no further testing was required.

The results of the study were summarised as follows:

Mortality: No mortality was observed at dose level of 2 000 mg/kg bw.

Clinical Observations: All animals were symptom-free during the 14-day observation period at a dose level of 2 000 mg/kg bw.

Body Weight and Body Weight Gain: There were no effects on body weights or body weight gains that could be attributed to treatment with the test material.

Necropsy: There was no evidence of the macroscopic observations in the animals dosed at 2 000 mg/kg bw and terminated on Day 14.

Under the conditions of this study, the acute oral LD50 value of the test material was found to be above 2 000 mg/kg bw in female Crl:WI rats.

According to the GHS criteria, the test material can be ranked as "Unclassified" for acute oral exposure.