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Description of key information

Key study: Test method OECD 420. GLP study. The substance was classified as category 4 according to CLP Regulation (EC) no. 1272/2008 since the dose of 2000 mg/kg bw caused death in the sighting study. No effects were observed at 300 mg/kg bw (main study).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 June 2014 - 1 July 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test method according to OECD Guideline 420. GLP study.
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Experimental Medicine Centre at the Medical University in Białystok
- Age at study initiation: 11 week-old
- Weight at study initiation: average body weight: 202.3 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): 19-25ºC
- Humidity (%): 40-85%
- Air changes (per hr): 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
Corn oil 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. The oil suspension of the test item was prepared on the administration day (shortly before the administration).
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 signs of toxicity were stated, the dose of 300 mg/kg b.w. was selected to be used in the main study.
Doses:
Sighting study: 300 and 2000 mg/kg bw
Main study: 300 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.
Preliminary study:
Clinical signs: No signs of toxicity were stated and the animal survived the experiment after administration of 300 mg/kg bw. Following the administration of the 2000 mg/kg bw test item, signs of toxicity were stated. The rounded back, a distinct decrease in locomotor activity, a decreased rate of reaction to sound stimuli, and a bristled coat were stated on the first day of the experiment. It died on the second day of the experiment.
Body weight: During the 14-day experiment, body weight gain was stated in the animal (dose of 300 mg/kg bw).
Gross examinations: No pathological changes were observed in both animals.
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.
Interpretation of results:
Category 4 based on GHS criteria
Remarks:
Migrated information
Conclusions:
The substance was classified as category 4 according to CLP Regulation (EC) no. 1272/2008 since the dose of 2000 mg/kg bw caused death in the sighting study. No effects were observed at 300 mg/kg bw (main study).
Executive summary:
The acute oral toxicity study based on a fixed dose method was performed according to OECD Guideline 420 (GLP study). The experiment commenced with a sighting study in which the test item was first administered to a female rat at a single dose of 300 mg/kg bw and then, since no signs of toxicity were observed, to a second rat at a single dose of 2000 mg/kg bw. The rounded back, a distinct decrease in locomotor activity, a decreased rate of reaction to sound stimuli, and a bristled coat were stated. It died on the second day of the experiment. Based on these results, four animals used in the main study were given the test item at a dose of 300 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 substance was classified as category 4 according to CLP Regulation (EC) no. 1272/2008 since the dose of 2000 mg/kg bw caused death in the sighting study.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
300 mg/kg bw
Quality of whole database:
Klimisch score = 1. GLP study.

Additional information

Key study: The acute oral toxicity study based on a fixed dose method was performed according to OECD Guideline 420 (GLP study). In the sighting study the test item was first administered to a female rat at a single dose of 300 mg/kg bw and then, since no signs of toxicity were observed, to a second rat at a single dose of 2000 mg/kg bw. The rounded back, a distinct decrease in locomotor activity, a decreased rate of reaction to sound stimuli, and a bristled coat were stated. It died on the second day of the experiment. Based on these results, four animals used in the main study were given the test item at a dose of 300 mg/kg bw. No signs of toxicity were observed. The substance was classified as category 4 according to CLP Regulation (EC) no. 1272/2008 since the dose of 2000 mg/kg bw caused death in the sighting study.


Justification for selection of acute toxicity – oral endpoint
Only one study available.

Justification for classification or non-classification

Based on the available results (discriminating dose of 300 mg/kg bw), the substance is classified for Acute Toxicity Category 4, according to CLP Regulation (EC) no. 1272/2008.