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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
26 May - 17 December, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
not applicable
Remarks:
Due to the short term nature of the study, no analysis was carried out to determine the homogeneity, concentration or stability of the test item formulation. This exception is considered not to affec the integrity or validity of the study.
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
not applicable
Remarks:
Due to the short term nature of the study, no analysis was carried out to determine the homogeneity, concentration or stability of the test item formulation. This exception is considered not to affec the integrity or validity of the study.
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure

Test material

Reference
Name:
Unnamed
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Physical state/Apparence : pale yellow powder
- Source and lot/batch No.of test material: 55503625
- Expiration date of the lot/batch: 28 April 2019
- Purity test date: 100%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark
- Stability under test conditions:
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: the test item was prepared as a suspension in arachis oil BP.
The arachid oil BP was used because the test item dod not dissolve/suspend in distilled water.

Test animals

Species:
rat
Strain:
Wistar
Remarks:
RccHan : WIST
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
- Females nulliparous and non-pregnant
- Age at study initiation: 8-12 weeks of age
- Fasting period before study: an overnight before dosing and for approximately 3 to 4 hours after dosing
- Housing: animal were foused in groups of up to four in suspended solid-floor polypropylene cages surnished with woodflakes
- Diet : ad libitum
- Water : ad libitum
- Acclimation period: 5 days
- Body Weight: the body weight variation did not exceed +/- 20% of the mean body weight at the stard of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25
- Humidity (%): 30 to 70
- Air changes (per hr): 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12/ 12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
Using available information on the toxicity ot the test item, 50 mg/kg was chosen as the starting dose
A single animal was treated with 50mg/kg.
In the absence of mortality at this dose level, an additional animal was treated with 300 mg/kg.
In the absence of mortality at this dose level, an additional animal was treated with 2000 mg/kg.
Due to mortality at the dose level of 2000 mg/kg, an additional group of 4 animals was treated at 300 mg/kg.
Doses:
50, 300, 2000 mg/kg
No. of animals per sex per dose:
1 animal for 50 mg/kg and 2000 mg/kg
5 animal for 300 mg/kg
Control animals:
no
Details on study design:
- Duration of clinical observation period following administration: 30 min, 1, 2 and 4 hours after dosing and the daily for up to 14 days.
- Frequency of weighing: day 0 and on day 7 and 14 or at death.
- Frequanecy of morbidity and mortality : twice daily, early and late during normal working day, and once daily at weekends and public holidays.
- Euthanasia : animals were killed by cervical dislocation
- Necropsy of survivors performed: yes for all animals

Evaluation of data included identification of the number of animals that died during the study (or that were killed for humane reasons), and determination of the nature, severity, onset and duration of the toxic effects. If possible, the signs of evident toxicity were described.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
At 50 mg/kg: no mortalilty
At 300 mg/kg: no mortality
At 2000 mg/kg: the animal was killed for humane reasons, 2 hours after dosing, due to the occurence of clinical signs of toxicity that approached the severity
limit set forth in th UK Home Office Project License.
Clinical signs:
At 50 mg/kg: Hunched posture and ataxia were noted during the day of dosing. The animal appeared normal 1 day after dosing.

At 300 mg/kg: No signs of systemic toxicity during the observation period.

At 2000 mg/kg: Signs of systemic toxicity noted were loss of righting reflex, decreased respiratory rate, labored respiration, pallor of the extremities, pilo-
ereection, prostration ans hypothermia.
Body weight:
At 50 mg/kg and 300 mg/kg: The animals showed expected gains in body weight over the observation period.

At 2000 mg/kg: Not applicable
Gross pathology:
At 50 mg/kg and 300 mg/kg: No abnormalities were noted at necropsy.

At 2000 mg/kg: Abnormalities noted at necropsy were patchy pallor of the liver, dark kidneys and clear liquid present in the stomach.
Other findings:
NA

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Executive summary:

An acute oral toxicity in the Wistar strain rat (A. Sanders, 2017, Envigo) has been performed, according the 420 OECD guideline in order to assess the LD50 of the test item.

Following a sighthing test at dose levels of 50, 300 and 2000 mg/kg, a further group of four fasted females was given a single oral dose of test item, as a suspension in arachis oil BP, at a dose level of 300 mg/kg. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

At 2000 mg/kg, animal was killed because of evident sign of systemic toxicity, confirmed by necropsy.

At 50 and 300 mg/kg, no mortality, no sign of systemic toxicity (also in necropsy) were observed.

Therefore the LD50 ot the test item in the female Wistar strain rat is estimated to be in the range of 300 -2000 mg/kg.