Glycine, N-acetyl-N-[3-(trifluoromethyl)phenyl]valyl-, ethyl ester

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 13 August 2002 to 30 December 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

17th July 1992

Deviations:

yes

Remarks:

One animal of the main test received by mistake a dose-volume of 2.5 mL/kg, instead of 2.05 mL/kg (dose received of 2475 mg/k. This minor deviation was not considered to have compromised the validity or integrity of the study.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Version / remarks:

31st July 1992

Deviations:

yes

Remarks:

One animal of the main test received by mistake a dose-volume of 2.5 mL/kg, instead of 2.05 mL/kg (dose received of 2475 mg/k. This minor deviation was not considered to have compromised the validity or integrity of the study.

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Glycine, N-acetyl-N-[3-(trifluoromethyl)phenyl]valyl-, ethyl ester

EC Number:

609-496-9

Cas Number:

379685-94-6

Molecular formula:

C18H23F3N2O4

IUPAC Name:

Glycine, N-acetyl-N-[3-(trifluoromethyl)phenyl]valyl-, ethyl ester

Test material form:

solid: particulate/powder

Details on test material:

- Other: ochre fine powder

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL- Storage condition of test material: room temperature

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Rj: SD (IOPS Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Janvier, Le Genest-Saint-IsIe, France.- Females (if applicable) nulliparous and non-pregnant: yes- Age at study initiation: approximately 6 weeks old- Weight at study initiation: 198 ± 9 g for the males and 164 ± 3 g for the females- Fasting period before study: ovenight period of approximately 18 hours before dosing, and during the 4 hours after administration of the test item. - Housing: animals were housed in polycarbonate cages with stainless steel lid (48 cm x 27 cm x 20 cm). Each cage contained one to seven animals of the same sex during the acclimation period and one rat (sighting test) or five rats of the same sex (main test) during the treatment period. Each cage contained autoclaved sawdust (SICSA, Alfortville, France). Sawdust was analysed by the supplier for composition and contaminant levels. - Diet: free access to A04 C pelleted diet (UAR, Villemoisson, Epinay-sur-Orge, France), except during the fasting period and the 4 hours following the test material administration. Each batch of food was analysed by the supplier for composition and contaminant levels. The diet formula was provided in the study report.- Water: Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum. Bacteriological and chemical analyses of water are performed regularly by extemal laboratories (include detection of possible contaminants (pesticides, heavymetals and nitrosamines)). - Acclimation period: at least 5 days ENVIRONMENTAL CONDITIONS - Temperature (°C): 22 ± 2°C - Humidity (%): 30 to 70% - Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air. - Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

methylcellulose

Remarks:

0.5% suspension of methylcellulose in purified water (prepared at the testing laboratory by reverse osmosis)

Details on oral exposure:

VEHICLE - Lot/batch no. (if required): 80K1020 (Sigma, Saint-Quentin-Fallavier, France) MAXIMUM DOSE VOLUME APPLIED: 10mLkgOn thé day of treatment, thé test item was ground using a mortar and pestle, then was prepared at thé chosen concentration in thé vehicle. DOSAGE FORM PREPARATION: On the day of treatment, the test item was ground using a mortar and pestle, and was then prepared at the chosen concentration in the vehicle. All preparations were made freshly on the morning of administration by the testing laboratory pharmacy and any unused material was discarded that same day.ADMINISTRATION OF THE TEST SUBSTANCE: The volume administered to each animal was adjusted according to body weight determined on the day of treatment.

Doses:

500 and 2000 mg/kg bw for the sighting test2000 mg/kg bw for the main test

No. of animals per sex per dose:

One female per dose level for the sighting test5 male and 5 females for the main test

Control animals:

no

Details on study design:

- Duration of observation period following administration: 7 days for the sighting test and 14 days for the main test - Frequency of observations and weighing: During the hours following treatment, animals were frequently observed. Then, at least once a day for 7 days (sighting test) or 14 day (main test) . All rats were weighted just before administration of the test item on day 1, and then on days 8 and 15.The body weight gain of the treated animals was compared to that of CIT control animals with a similar initial body weight.- Sacrifice: by CO2 asphyxiation - Necropsy of survivors performed: yes, as soon as possible after death. After opening the thoracic and abdominal cavities, a macroscopic examination of the main organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities) was performed. No organ samples were taken.

Statistics:

not applicable

Results and discussion

Preliminary study:

Dyspnea and piloerection were observed on day 1 in the animal given 500 mg/kg bw.No clinical signs and no mortality were observed at the 2000 mg/kg dose level.

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality, no significant clinical signs and no macroscopic findings

Mortality:

No mortality was observed.

Clinical signs:

other: Piloerection was observed on day 1 in all animals.

Gross pathology:

Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.

Other findings:

None.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study, the oral LD50 of the test material is higher than 2000 mg/kg body weight without mortality, significant clinical signs or macroscopic findings. Thus, the test material is not classified as hazardous for acute oral toxicity according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) criteria.

Executive summary:

This GLP-compliant study was performed to assess the acute oral toxicity of the test material, according to OECD Guideline 420 (dated July 17th, 1992) and EU method B.1 bis of the E.E.C. directive n° 92/69 (dated July 31st, 1992).

Material and methods

The test material -prepared in a 0.5% suspension of methylcellulose- was administered by gavage to a group of 10 Sprague Dawley rats (5 males and 5 females) at the dose of 2000 mg/kg body weight. This dose was chosen based on the results of a preliminary test (sighting test) where the doses of 500 and 2000 mg/kg body weight were administered to female rats (one per dose group).

Animals of the main test were monitored at least once a day for 14 days. All rats were weighted just before administration of the test item on day 1, and then on days 8 and 15. Necropsies were performed on all the animals.

Results

No mortalities or significant clinical signs were recorded during the study. Only piloerection was observed on day 1 in all animals. The body weight gain of the treated animals was not affected by the treatment with the test material. The macroscopic examination of the animals at the end of the study did not reveal any treatment-related change.

Conclusion

Under the experimental conditions of this study, the oral LD50 of the test material is higher than 2000 mg/kg body weight without mortality, significant clinical signs or macroscopic findings. Thus, the test material is not classified as hazardous for acute oral toxicity according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) criteria.