Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 8 March 2016 to 27 April 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Stabilisation: in water undergoes hydrolysis
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Purity: 92%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, humidity should be avoid.


Test animals

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 - 12 week old.
- Weight at study initiation: For the preliminary study 236-245 g. For the main study an average body weight of 231 g.
- Fasting period before study: 19h
- Housing:plastic cages covered with wire bar lids. The dimensions of the cages were 58 x 37 x 21 cm (length x width x height). In the preliminary experiment, the animals were caged individually. In the main experiment, there were four animals in one cage.UV-sterilized wood shavings were used as bedding
- Diet: ad libitum
- Water: ad libitum
- Acclimation period:The animals were quarantined and observed daily for 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 – 25 °C
- Humidity (%): 30 – 51%
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

IN-LIFE DATES:
Preliminary experiment: The animal treated with 300 mg/kg bw from 30.03.2016 to 13.04.2016. The animal treated with 2000 mg/kg bw from 05.04.2016 to 19.04.2016.
Main experiment: From 08.04.2016 to 22.04.2016.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle:
For the preliminary experiment - 60 mg test item/1 ml oil suspension in one animal and 400 mg/1 ml oil suspension in a second animal.
For the main experiment: 400 mg/1 ml oil suspension
- Amount of vehicle (if gavage): 0.5 mL/100 g b.w. in the preliminary and main experiment
- Justification for choice of vehicle: Due to the low water solubility of the test item.
Doses:
Preliminary experiment: 300 mg/kg b.w.and 2000 mg/kg b.w.
Main experiment: 2000 mg/kg b.w.
No. of animals per sex per dose:
Preliminary experiment: 1 animal per sex per dose
Main experiment: 4 animals per sex per dose
The animal from the preliminary experiment which had received the dose of 2000 mg/kg b.w. was included in the main experiment.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The observation of all animals for morbidity and mortality was conducted twice a day or once a day (on days off) during the 14-day experiment. Body weights of the animals were determined on days 0, 7, and 14.
- Necropsy of survivors performed: yes
- Other examinations performed:
Clinical signs: evaluation of general condition of the animals and detailed clinical observations (locomotor system, behaviour, reactions to stimuli, skin and hair, eyes and eyelides, respiratory system, urinary system, digestive system, reproductive system).
Gross examinations: observation of the external body surface, all natural apertures, and the cranial, thoracic, and abdominal cavities with their content.
Statistics:
Not performed

Results and discussion

Preliminary study:
No signs of toxicity were stated on animals treated with 300 mg/kg b.w.and 2000 mg/kg b.w. Both animals survived the experiment. Body weight gain was observed in both animals but gross examinations did not reveal any pathological changes. According to these results, in the main experiment was administered a dose of 2000 mg/kg b.w.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
The animals survived the experiment.
Clinical signs:
No signs of toxicity were stated.
Body weight:
During the second week of the experiment, body weight loss in two females was observed. During the 14-day experiment body weights of all animals increased.
Gross pathology:
Gross examinations did not reveal any pathological changes in the examined animals.

Any other information on results incl. tables

Body weights of the animals (g) – overall list.

Dose

(mg/kg b.w.)

Animal

number

Day of the experiment/

Body weight (g)

Body weight

gain(g)

(From 0 to 14)

0

7

14

300

1*

236

261

264

28

  

2000

1*

245

266

269

24

  

2000

2

231

253

253

22

 

2000

3

227

252

247

20

  

2000

4

238

265

259

21

  

2000

5

228

259

264

36

* the female from the preliminary experiment.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The median lethal dose (LD50) of the test item on female rats is greater than 2000 mg/kg b.w.
Executive summary:

The acute oral toxicity of the test item was determined on female Wistar rats in accordance to the OECD Guideline 420 with GLP. A preliminary experiment was conducted in which the test item at a single dose of 300 mg/kg b.w. was administered to one animal. Since no evident toxicity was observed in dose 300 mg/kg b.w., the test item at a single dose of 2000 mg/kg b.w. was administered to the second animal. No signs of toxicity or mortality were observed on both animals. On the grounds of the preliminary experiment results, four animals used in the main experiment were given the test item at a dose of 2000 mg/kg b.w. The test item in the form of an oil suspension in a volume of 0.5 mL/100 g b.w. was administered with a metal stomach tube. 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 and body weights of the animals were determined on day 0, 7 and 14. During the 14-day experiment body weights of all animals increased. No signs of toxicity were stated and the animals survived the experiment. Gross examinations did not reveal any pathological changes in the examined animals. On the grounds of the study results LD50 of the test item is greater than 2000 mg/kg b.w.