P-1401

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

4 August 2015 - 25 August 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Experimental Medicine Centre at the Medical University in Białystok
- Age at study initiation: 10 week-old (preliminary test); 11 week-old (main test)
- Weight at study initiation: average body weight: 207-217 g
- Fasting period before study: about 19 hours before the administration of the test item, restored 3 hours after the administration
- Housing: plastic cages covered with wire bar lids, 58 x 37 x 21 cm, with UV-sterilized wood shavings as bedding.
- Diet (e.g. ad libitum): ad libitum, "Murigran" standard granulated laboratory food.
- Water (e.g. ad libitum): ad libitum, tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-26ºC
- Humidity (%): 40-87%
- Air changes (per hr): 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

(distilled water)

Details on oral exposure:

VEHICLE: Distilled water was introduced to a beaker containing 300 mg (300 mg/Kg bw dose) or 2000 mg (2000 mg/kg bw dose) of the test item to obtain a solution in a volume of 5 mL. Then, the test was carefully mixed with distilled water.
Concentration in vehicle: 60 mg/ml (300 mg/kg bw dose); 400 mg/ml (200 mg/kg bw dose).

MAXIMUM DOSE VOLUME APPLIED: 0.5 mL/100 g bw

RATIONALE FOR SELECTION OF THE STARTING DOSE: The starting dose for the sighting study was selected from the fixed dose levels of 5, 50, 300 and 2000 mg/kg bw. Since no data were available, the sighting study commenced with the administration of the test item at a dose of 300 mg/kg bw to one female rat. Considering the fact that no evident toxicity was produced, the test item at a single dose of 2000 mg/kg bw was administered to the next animal. Since no signs of toxicity were stated, the dose of 2000 mg/kg b.w. was selected to be used in the main study.

Doses:

Sighting study: 300 and 2000 mg/kg bw
Main study: 2000 mg/kg bw

No. of animals per sex per dose:

2 female in the sighting study.
4 females in the main study.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: on days 0 (directly before the administration of the test item), 7, and 14 (before euthanasia).
- Necropsy of survivors performed: yes
- Other examinations performed:
General condition: observation of all animals for morbidity and mortality: twice a day or once a day (on days off) during the 14-day experiment.
Detailed clinical observations: on the day of the test item administration (day 0), i.e. 10, 30, and 60 minutes after the administration and then at hourly intervals up to the 5th hour after the administration. From the 1st to the 14th day of the experiment, the detailed clinical observations were performed once a day.
Gross examinations: After the 14-day observation period, all animals were euthanized by intraperitoneal administration of morbital at a dose of 200 mg/kg bw and subjected to gross examinations. The detailed gross examinations comprised the observation of the external body surface, all natural apertures, and the cranial, thoracic, and abdominal cavities with their contents.

Results and discussion

Preliminary study:

Clinical signs: No signs of toxicity were stated and the animals survived the experiment after administration of 300 mg/kg bw and 2000 mg/kg bw.
Body weight: During the 14-day experiment, body weight loss was observed in one female (dose of 300 mg/kg bw).
Gross examinations: No pathological changes were observed in both animals.

Effect levels

Key result

Sex:

female

Dose descriptor:

discriminating dose

Effect level:

300 mg/kg bw

Based on:

test mat.

Mortality:

All animals survived the experiment.

Clinical signs:

No signs of toxicity were found.

Body weight:

During the 14-day experiment, body weight gain was found in all animals.

Gross pathology:

No lesions were found in the animal.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

CLP Implementation.

Conclusions:

No effects were observed in rats after the oral exposure of 2000 mg/kg bw (highest dose tested).

Executive summary:

The acute oral toxicity study based on a fixed dose method was performed according to OECD Guideline 420 (GLP study). In the preliminary test a female rat was orally exposed to a single dose of 300 mg/kg bw and then, since no signs of toxicity were observed, a second rat was exposed to a single dose of 2000 mg/kg bw. Since no effects were observed and all rats survided, four animals used in the main study were given the test item at a dose of 2000 mg/kg bw. After the administration of the test item, the animals were observed for 14 days. General and detailed clinical observations were conducted daily during the entire experiment. Body weights of the animals were determined. After the 14-day observation period, the animals were euthanized and subjected to a necropsy and a detailed gross examination. No signs of toxicity were found and all animals survived the experiment. During the 14-day experiment, body weight gain was found in the animals. Regarding the gross examinations, no lesions were found in the animals. The test item was beyond categorization according to CLP Regulation (EC) No. 1272/2008.