Ethyl 2-methylvalerate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 March to 19 Apr 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed accoding to GLP, OECD Guidelines followed and no deviations reported

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl 2-methylvalerate

EC Number:

254-384-1

EC Name:

Ethyl 2-methylvalerate

Cas Number:

39255-32-8

Molecular formula:

C8H16O2

IUPAC Name:

ethyl 2-methylpentanoate

Test material form:

not specified

Details on test material:

- Name of test material (as cited in study report): EMV (EMV correspond to Ethyl 2-methylvalerate)
- Expiration date of the lot/batch: 25 Jul 2014
- Storage condition of test material: Immediately upon receipt, the test item was registered, then stored at room temperature in accordance with the Sponsor’s instructions. The complete description of the chemical and physical properties of the test item including stability is the responsibility of the Sponsor.
- Other:
- Supplier: Toyo Gosei Co., Ltd.
- Intended use: Ind. Chemical
- On 12 Mar 2012, one sample of test item was received in vial labelled “Ethyl 2-methylvalerate(EMV) Batch No. H3-G-26”. The study monitor confirm that this test item correspond to “EMV Batch No. H3-G-26’’, name used in the study report.
- Handling instructions for test item: General safety procedures as appropriate for handling of chemicals of unknown hazard potential were applied. For further details about safety, see the material safety data sheet supplied with the test item by the Sponsor.
- Remaining test item: The remaining test item, except the sample to he archived, (in case of archiving of test item) will be sent to Laboservice, route de Ia Centrale, 69702, Givors Cedex, where it will he destroyed by incineration, under the responsibility of CERB.
- The certificate of analysis of test item is presented in Appendix A, page 28 of attached report.

Test animals

Species:

rat

Strain:

other: SPF (Specific Pathogen Free) Sprague-Dawley -Crl:OFA (SD)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratoires France - Domaine des Oncins, 69210 L’Arbresle Cedex.
- Age at study initiation: 9 weeks at the time of administration.
- Weight at study initiation: Within ± 20% of the mean weight of any previously dosed animals.
- Housing: Daily observations were undertaken at the time of delivery of the animals and during the period of acclimatisation. Animals were housed in groups of 5 at maximum, in cages of standard dimensions with sawdust bedding.
- Diet (e.g. ad libitum): Feeding: RM1 (E)-SQC SDS/DIETEX feed (quality controlled) was available ad libitum except during the fasting experimental period. The certificate of analysis concerning this feed product is included in Appendix B, page 30 of attached report. The criteria for acceptable levels of contaminant, in the feed supply were thin the limits of the analytical specifications established by the diet manufacturer.
- Water (e.g. ad libitum): Drinking water: Drinking water was available ad libitum in polvcarbonate bottles with a stainless steel nipple. A specimen of water is obtained approximately every 6 months and sent to the LAEASE Region Sud Est - 5, avenue Achille Maureau – B. P. 95 - 84703 Sorgues Cedex - France, for analysis. The certificate of analysis are included in Appendix C, page 32 of attached report. The criteria for acceptable levels of contaminants in the water supplied were within the limits of the analytical specifications.
- Acclimation period: Acclimatisation: Five days before treament in the laboratory animal house where the experiment took place.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 - No deviations outside of the temperature or hygrometry ranges were reported by the Study Director according to the SOP 5.36.
- Humidity (%): Relative humidity between 45% and 65% (except during the cleaning slot)
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): The artificial day/night cycle involved 12 hours light and 12 hours darkness with light on at 7.30 am.

- Other:
- Identification: Animals were identified individually by marking the tail with a felt tip marker.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Justification for choice of vehicle: Corn oil was chosen after the solubility trial, on the basis of the physico-chemical characteristics of the test item
- Lot/batch no. (if required): SIGMA, Batch No. MKBF8603V, Expiry date: 5 Mar 2017

MAXIMUM DOSE VOLUME APPLIED: 10mL/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: not provided

Doses:

2000 mg/kg.

No. of animals per sex per dose:

Female: 6 animals with 3 animals per step.

Control animals:

no

Details on study design:

Examination:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were weighed on the following days:
- on D1, day of administration
- on D4, D7, D10 and D14 during the study
- on D15, day of necropsy (not exsanguinated)
- Necropsy of survivors performed: yes - All animals surviving to the end of the 14-day monitoring period were euthanased on D15 by subtotal exsanguination, after isoflurane inhalation. All animals were subjected to gross necropsy and their organs (brain, liver, spleen, kidneys, stomach, intestines, gonads/reproductive tact, lungs and heart) were examined macroscopically.

- Other examinations performed:
- Mortality - Morbidity: Mortality was recorded twice a day, i.e. in the morning and at the end of the working day.
- Clinical signs:
- General examination: Animals were examined clinically once before dosing and twice on the day of treatment (30 minutes ± 2 minutes post-dose and then again between 3 and 4 hours post-dose). Thereafter they were examined clinically at least once a day for 14 days. The general disposition, behaviour and activity were observed (See Section 1, page 10 of attached report).
- Functional and neurobehavioural tests: On D1 (30 minutes ± 1 minutes post-dose), on D7 and on D14, animals were observed according to a standardised observation battery for general clinical signs, neurobehavioural, neurovegetative or psychotropic signs or neurotoxic effects. The method is based on the Irwin screen* modified by suppressing the graduation of intensity of clinical signs. Animals were observed individually in a cage with out sawdust in a quiet room. Clinical signs were observed according to Table 1.1, page 21 of attached report.
- Food and water consumption, fasted periods: Food and water consumption were not measured. Animals were fasted during the night before treatment and remain deprived of food for 3 to 4 hours post-dose.

- Pathology: Euthanasia were performed by CERB under the responsibility of P. Pradeau, DVM, AnatomopathoIogist.

- Method of administration:
- Timing, frequency and duration of administration: EMV was administered to animals deprived of food since the previous day. It was administered to the animals as a single dose, by gavage, using a cannuIa of appropriate size.
- Volume to be administered: The dose volume was 10 mL/kg.

* Irwin S. Comprehensive observational assessment: A ystematic quantitative procedure fo assessing the behavioural and physiological state of the mouse. Psychopharmacologica. 1968; 13:222-257.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Remarks on result:

other: No deaths and no evidence of toxicity at 2000 mg/kg

Mortality:

MORTALITY: No mortality occurred during the study.

Clinical signs:

other: CLINICAL FINDINGS: There was no clinical sign during the study.

Gross pathology:

MACROSCOPIC FINDINGS: Individual data are presented in Table 2.5, page 26 of attached report. No findings were seen at necropsy.

Any other information on results incl. tables

DISCUSSION: Since the body weight change was minimal (at maximum -2%) and because body weight increased or remained stable in three other animals between D7 and D10 and in all animals on D14, this change was not attributed to an effect of the test item.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the experimental conditions adopted, EMV administered once by oral route in the Sprague Dawley rat at 2000 mg/kg did not induce any sign of toxicity.

Executive summary:

The aim of this study was to determine the acute toxicity of EMV at 2000 mg/kg after single dose administration by the oral (gavage) route in the rat.

 

Experimental procedure:

- 6 animals were treated at 2000 mg/kg.

- Mortality was recorded twice a day for 14 days. Clinical observations were performed before the dosing then daily. Functional and neurobehavioural tests were performed on D1, D7 and D14.

- Body weight was recorded on D1, D4, D7, D10 and D14.

- Animals were subjected to gross necropsy on D15.

 

Results:

- Mortality: No mortality occurred during the study.

- Clinical signs: There was no clinical sign during the study.

- Body weight: There was no treatment-related change in body weight gain.

- Macroscopic examination: No findings were seen at necropsy.

 

Conclusion:

Under the experimental conditions adopted, EMV administered once by oral route in the Sprague Dawley rat at 2000 mg/kg did not induce any sign of toxicity.