Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From July 01-29, 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted in compliance with OECD Guideline 420 without any deviation.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

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
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Programme (inspected on March 12 to 14, 2014/ signed on May 12, 2014)
Test type:
fixed dose procedure
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): (E)-1-(2,6,6-trimethyl-1-cyclohexen-1-yl)pent-1-en-3-one
- Physical state: Pale yellow liquid
- Storage condition of test material: Stored at room temperature in the dark.

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 137-180 g
- Fasting period before study: Animals were fasted for overnight period before dosing and for approximately 3-4 h after dosing.
- Housing: Animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet: Food (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK), ad libitum
- Water: Mains drinking water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-25 °C
- Humidity: 30-70 %
- Air changes: 15 changes / h
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: From: July 01, 2014 To: July 29, 2014

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Arachis oil BP for 300 mg/kg bw; no vehicle (unchanged) for 2000 mg/kg bw
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 mg/mL
- Justification for choice of vehicle: Test material at 300 mg/kg bw did not dissolve/suspend in distilled water and therefore solution of test material in arachis oil BP was prepared.

DOSAGE PREPARATION (if unusual): Test material at 2000 mg/kg bw dose level was used as supplied. 300 mg/kg bw dose level was freshly prepared as solution in arachis oil BP. Test material was formulated within 2 h before dosing.

DOSE VOLUME APPLIED: 10 mL/kg bw for 300 mg/kg bw; 2.15 mL/kg bw for 2000 mg/kg bw (specific gravity: 0.933)
Doses:
- Sighting study: 300 and 2000 mg/kg bw
- Main study: 2000 mg/kg bw
No. of animals per sex per dose:
- Sighting study: 1 female/dose
- Main study: 5 females/dose (1 animal included from sighting study)
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 h after dosing and then daily for 14 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; at the end of the observation period, animals were killed by cervical dislocation and subjected to gross necropsy.
Statistics:
None

Results and discussion

Preliminary study:
- No mortality or clinical signs were observed at 300 mg/kg bw.
- The animal showed a body weight gain over the observation period that was considered to be within the historical range for this strain.
- No abnormalities were noted at necropsy.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed.
Mortality:
- No mortality was observed at 2000 mg/kg bw.
Clinical signs:
- Hunched posture was noted 4 h after dosing in three animals. No other signs of systemic toxicity noted in two animals.
Body weight:
- Animals showed body weight gains over the observation period that were considered to be within the historical range for this strain.
Gross pathology:
- Epithelial sloughing of the non-glandular epithelium of the stomach was noted at necropsy of one animal. No abnormalities were noted at necropsy of four animals.
Other findings:
None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Rat Oral LD50 (females) > 2000 mg/kg bw
Executive summary:

In an acute oral toxicity study performed according to OECD Guideline 420 and in compliance with GLP, groups (5 females/dose) of Wistar (RccHan™:WIST) rats were given a single oral (gavage) dose of test material at 2000 mg/kg bw. Animals were then observed for mortality, clinical signs and bodyweights for 14 days and at the end of the study the surviving animals were sacrificed for macroscopic examination. Sighting study was conducted at the dose levels of 300 and 2000 mg/kg bw (one female/dose) to determine the dose for main study.

In the sighting study, no mortality or clinical signs were observed at 300 mg/kg bw. The animal showed a body weight gain over the observation period that was considered to be within the historical range for this strain. No abnormalities were noted at necropsy. In the main study, no mortality was observed at 2000 mg/kg bw. Hunched posture was noted 4 h after dosing in three animals. There were no other signs of systemic toxicity noted in two animals. Animals showed body weight gains over the observation period and that were considered to be within the historical range for this strain. Epithelial sloughing of the non-glandular epithelium of the stomach was noted at necropsy of one animal. No abnormalities were noted at necropsy of four animals.

 

Rat Oral LD50 (females) > 2000 mg/kg bw

 

Under the test conditions, the test material is not classified according to the annex VI of the Regulation EC No. 1272/2008 (CLP).

This study is considered as acceptable and satisfies the requirement for acute oral toxicity endpoint.