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Administrative data

acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24-07-2018 to 21-08-2018
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.
Justification for type of information:
Information as to the availability of the in vivo study is provided in 'attached justification'.

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guidelineopen allclose all
according to guideline
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
according to guideline
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
GLP compliance:
yes (incl. QA statement)
inspected: July 2017 ; signature: November 2017
Test type:
fixed dose procedure
Limit test:

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
Cas Number:
Molecular formula:
Test material form:
Details on test material:
- Physical state: Liquid
- Storage condition of test material: approximately 4ºC, in the dark, under nitrogen
- Other: colourless to pale yellow liquid

Test animals

Details on test animals or test system and environmental conditions:
- Source: Recognised supplier
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 - 12 weeks.
- Weight at study initiation: 150-162 g (300 mg/kg including sighting test; sentinel); 160-175 g (2000 mg/kg sighting test; sentinel); The weight variation did not exceed ±20% of the mean weight in the definitive test.
- Fasting period before study: Overnight before dosing and three to four hours after dosing.
- Housing: Group housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum (except for fasting period).
- Water (e.g. ad libitum): ad libitum (except for fasting period)
- Acclimation period: At least 5 days.

- Temperature (°C): 19 - 25
- Humidity (%): 30 - 70%
- Air changes (per hr): > 15 air changes per hour
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES: From: To: 24-07-2018 to 21-08-2018

Administration / exposure

Route of administration:
oral: gavage
arachis oil
Details on oral exposure:
- Concentration in vehicle: The specific gravity was determined and used to calculate the appropriate dose volume for the required dose level. For the purpose of the 300 mg/kg dose level the test item was freshly prepared, as required, as a solution in arachis oil BP. Arachis oil BP was used because the test item did not dissolve/suspend in distilled water. For the purpose of the 2000 mg/kg dose level, the test item was used as supplied. The 300 mg/kg bw and 2000 mg/kg bw dose levels were treated stepwise. Singularly and in the absence of mortality or evident toxicity a further group of 4 was tested in the appropriate dose level.
The test item was formulated at concentrations of 30 mg/mL or dosed as supplied according to specific gravity of the test item. Formulations were prepared on the day of dosing.
- Amount of vehicle (if gavage): Test Item dose volume was 10 mL/kg (of bodyweight) in Arachis Oil BP or dosed as supplied at 2.35 mL/kg
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed.
- Lot/batch no. (if required): See full study report.
- Purity: BP grade.

MAXIMUM DOSE VOLUME APPLIED: 300 mg/kg bw dose level: 10 mL/kg of test item in vehicle (Arachis Oil BP) ; 2000 mg/kg bw dose level: 2.35 mL/kg of test item as supplied.

DOSAGE PREPARATION (if unusual): Not applicable. The test item was prepared in the vehicle. It was administered to the animals under a volume of 10 mL/kg in Arachis Oil BP (300 mg/kg bw) or 2.35 mL/kg as supplied (2000 mg/kg bw).

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose based on guideline recommendations.
300 mg/kg bw (initial sighting test and main study)
2000 mg/kg bw (initial sighting test and main study)
No. of animals per sex per dose:
300 mg/kg bw in Arachis Oil vehicle (starting dose) : 1 (sighting study) and 4 (main study) as applicable.
2000 mg/kg bw: 1 (sighting study) and 4 (main study) as applicable; total 5 per dose - based on guideline specified sequential testing strategy
Control animals:
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made 0.5, 1, 2, and 4 hours after dosing and then daily for fourteen days. Morbidity and mortality checks were made twice daily. Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: yes

Results and discussion

Effect levels
Dose descriptor:
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
300 mg/kg bw (sentinel and definitive test): No mortality
2000 mg/kg bw (sentinel and definitive test): Two mortalities on day 2 (2/5)
Clinical signs:
other: 300 mg/kg bw (sentinel and definitive test): No signs of systemic toxicity were noted. 2000 mg/kg bw (sentinel and definitive test): Hunched posture on day of dosing and up to day 4 (3/5), ataxia (1/5) which ceased one day after dosing. Within the mortali
Gross pathology:
300 mg/kg bw (sentinel and definitive test): No abnormalties were noted at necropsy.
2000 mg/kg bw (sentinel and definitive test): No abnormalties were noted at necropsy in two survivors. One survivor had ulceration of the non-glandular region of the stomach at the end of the observation period. Within the mortalities: patchy pallor of the liver was noted at necropsy

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Criteria used for interpretation of results: EU
Under the conditions of this study, the oral LD50 was estimated to be in the range of 300 - 2000 mg/kg bw in female Wistar rats.
Executive summary:

The study was performed according to OECD TG 420 and EU Method B.1 bis Acute Toxicity (Oral) and in accordance with GLP to assess the acute oral toxicity of the test material following a single oral administration in the female Wistar strain rat by the fixed dose method. The test item was administered by oral gavage in an initial sighting study at 300 mg/kg bw in a solution in arachis oil BP and following an absence of toxicity then at 2000 mg/kg bw unchanged. Subsequently, at a further group of four fasted females was given a single oral dose of test item, at a dose level of 300 mg/kg body weight and/or 2000 mg/kg bw depending on the findings at the preceding dose level.Two mortalities were seen at a dose level of 2000 mg/kg on Day 2. There were no deaths noted at a dose level of 300 mg/kg. Signs of systemic toxicity noted at a dose level of 2000 mg/kg in the mortalities during the study were loss of righting reflex, lethargy, decreased respiratory rate and laboured respiration. Within survivors hunched posture (three of five) and ataxia was observed (in one of five at this dose level). All survivors gained bodyweight. There were no clinical signs of systemic toxicity at 300 mg/kg bw and all animals showed expected gains in bodyweight over the study period.Patchy pallor of the liver was noted at necropsy in the mortalities at a dose level of 2000 mg/kg. Ulceration of the non-glandular region of the stomach was noted at necropsy of one survivor. At 300 mg/kg bw there was no abnormalities noted. Under the conditions of this study, the oral LD50 was estimated to be in the range of 300 - 2000 mg/kg bw in the female Wistar rat.