Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008-02-27 to 2008-03-26
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Version / remarks:
, 2008
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Version / remarks:
, adopted 2001-12-17
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
signed 2007-10-15
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
- EC name: Acetic-acid, 2- (ethylamino) -2-oxo, (1R,2S,5R) -5-methyl- 2- (1-methylethyl)- cyclohexyl ester
- Molecular formula: C14H25NO3
- Molecular weight: 255.36g/mol
- Physical state: clear colourless extremely viscous liquid
- Storage condition of test material: room temperature in the dark

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, UK
- Age at study initiation: eight to twelve weeks of age
- Weight at study initiation: 213 - 234 g
- Fasting period before study: an overnight fast immediately before dosing and for approximately three to four hours after dosing
- Housing: the animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet (ad libitum, exception see "Fasting period before study" above): Certified Rat and Mouse Diet
- Water (ad libitum): mains drinking water
- Acclimation period: at least five days

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

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
SIGHTING STUDY / MAIN STUDY
In absence of data regarding the toxicity of the test material 300 mg/kg was chosen as the startind dose. A single animal was treated as follows:
dose level 300 mg/kg; concentration 30 mg/ml; dose volume 10 ml/kg.
In the absence of toxicity at a dose level of 300 mg/kg, an additional animal was treated as follows:
dose level 2000 mg/kg; concentration 200 mg/ml; dose volume 10 ml/kg.
In the absence of toxicity at a dose level of 2000 mg/kg, an additional group of 4 animals was treated as follows:
dose level 2000 mg/kg; concentration 200 mg/ml; dose volume 10 ml/kg.
A total of five animals were therefore treated at a dose level of 2000 mg/kg in the study.

The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

VEHICLE
- Justification for choice of vehicle: Arachis oil BP was used because the test material did not dissolve/suspend in distilled water.

DOSAGE PREPARATION: The test material was freshly prepared a a solution in arachis oil BP.
Doses:
300 mg/kg bw. (sighting study)
2000 mg/kg bw. (sighting study / main study)
No. of animals per sex per dose:
300 mg/kg bw. one female rat
2000 mg/kg bw. five female rats
Control animals:
no
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 bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: yes; at the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
Statistics:
not applicable

Results and discussion

Preliminary study:
DOSE LEVEL 300 mg/kg bw:
- There was no mortility.
- No signs of systemic toxicity were noted during the observation period.
- The animal showed expected gains in bodyweight over the observation period.
- No abnormalities were noted at necropsy.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths.
Clinical signs:
No signs of systemic toxicity were noted during the observation period.
Body weight:
All animals showed expected gains in bodyweight over the observation period.
Gross pathology:
No abnormalities were noted at necropsy.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the female Sprague-Dawley CD strain rat was estimated to be greater than 2000 mg/kg bw.
According to the EC-Commission directive 67/548/EEC and its subsequent amendments, the test substance is not classified as acute toxic via the oral route.
According to the EC-Regulation 1272/2008 and subsequent regulations, the test item is not classified as acute toxic via the oral route.