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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Between 21 April 2005 and 25 May 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report date:
2005

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
β,4-dimethylcyclohex-3-ene-1-propan-1-al
EC Number:
229-846-0
EC Name:
β,4-dimethylcyclohex-3-ene-1-propan-1-al
Cas Number:
6784-13-0
Molecular formula:
C11H18O
IUPAC Name:
3-(4-methylcyclohex-3-en-1-yl)butanal
Test material form:
liquid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, UK
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-12 weeks
- Fasting period before study: The rats were fasted overnight prior to dosing, with food being returned to the rats approximately 3-4 hours after the first dose
- Housing: The animals were housed in groups of three in suspended solid-floor polypropylene cages furnished with woodflakes
- Diet:ad libitum except during fasting
- Water: water ad libitum except during fasting
- Acclimation period: 5 days
- Microbiological status when known: 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.

ENVIRONMENTAL CONDITIONS
- Temperature: 19-25ºC
- Humidity: 30-70%
- Air changes: The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: The test material was administered orally undiluted for the 2000 mg/kg dose level and orally as a solution in arachis oil BP for the 300 mg/kg dose level.
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 mg/mL for 300 mg/kg
- Justification for choice of vehicle: Arachis oil BP was used because the test material did not dissolve/suspend in distilled water

MAXIMUM DOSE VOLUME APPLIED: 2.14 mL/kg for 2000 mg/kg; 10 mL/kg go 2000 mg/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Using all available information on the toxicity of the test material, 2000 mg/kg was chosen as the starting dose.
Doses:
300 mg/kg bw and 2000 mg/kg bw.
No. of animals per sex per dose:
6 females per dose
Control animals:
no
Details on study design:
The animals were observed for deaths or overt signs of toxicity 1/2, 1, 2 and 4 hours after dosing and subsequently once daily for up to fourteen days. Due to a technical error the observations of three animals treated at a dose level of 300 mg/kg on Day 2 were not performed. This was considered not to affect the purpose or integrity of the study.
Individual bodyweights were recorded prior to dosing and seven and fourteen days after treatment or at death. At the end of the observation period the surviving animals were killed by cervical dislocation. All animals were subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities for examination of major organs. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
One animal treated at a dose level of 2000 mg/kg was killed in extremis and one other animal at
this dose level was found dead one day after dosing. There were no deaths noted in animals treated at a dose level of 300 mg/kg.
Clinical signs:
other:
Body weight:
other body weight observations
Remarks:
The surviving animals showed expected gains in bodyweight over the study period.
Gross pathology:
Abnormalities noted at necropsy of the animal that died during the study were abnormally red lungs, dark liver and dark kidneys. No abnormalities were noted at necropsy of animals that were killed in extremis or at the end of the study.

Applicant's summary and conclusion

Interpretation of results:
other: not acute harmful according to EU CLP (EC 1272/2008 and its amendments)
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the female Sprague-Dawley CD strain rat was >2000 mg/kg bw
Executive summary:

The study was conducted to assess the acute oral toxicity of limoxal in accordance with the OECD Guideline 423 in compliance with GLP.


A group of three fasted females was treated with the test material at a dose level of 2000 mg/kg bw. This was followed by a further group of three fasted females at the same dose level. Based on the results from this dose level a further two groups of fasted females were treated at a dose level of 300 mg/kg bw. Dosing was performed sequentially. The test material was administered orally undiluted for the 2000 mg/kg dose level and orally as a solution in arachis oil BP for the 300 mg/kg dose level.  Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.


At 2000 mg/kg, one animal was found dead one day after dosing and one animal was killed in extremis at this time. Signs of systemic toxicity noted in animals treated at a dose level of 2000 mg/kg were hunched posture, ataxia, lethargy, pilo-erection, decreased respiratory rate, laboured respiration, loss of righting reflex, occasional body tremors, tiptoe gait and coma. Surviving animals treated at a dose level of 2000 mg/kg appeared normal two, three or six days after dosing. There were no signs of systemic toxicity noted in animals treated at a dose level of 300 mg/kg. The surviving animals showed expected gains in bodyweight over the study period. Abnormalities noted at necropsy of the animal that died during the study were abnormally red lungs, dark liver and dark kidneys. No abnormalities were noted at necropsy of animals that were killed in extremis or at the end of the study.


The acute oral median lethal dose (LD50) of the test material in the female Sprague-Dawley CD strain rat was >2000 mg/kg bw.