Reaction mass of Ethyl 3-(cyclohex-1-en-1-yl) propanoate and Ethyl 3-(cyclohex-2-en-1-yl) propanoate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

This study was conducted between 03 January 2017 and 30 January 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Reliability 1 is assigned because the study conducted according to OECD TG 420 in compliance with GLP, without deviations that influence the quality of the results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

EC No. 440/2008

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of Ethyl 3-(cyclohex-1-en-1-yl) propanoate and Ethyl 3-(cyclohex-2-en-1-yl) propanoate

EC Number:

949-881-1

Molecular formula:

C11H18O2

IUPAC Name:

Reaction mass of Ethyl 3-(cyclohex-1-en-1-yl) propanoate and Ethyl 3-(cyclohex-2-en-1-yl) propanoate

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Sponsor Batch No. NG342-12
- Expiration date of the lot/batch: 01 September 2017
- Purity test date: 07 September 2016
- Purity: 96.6%
- Appearance: clear colorless liquid
- Storage condition of test material: Approximately 4”C in the dark

Test animals

Species:

rat

Strain:

Wistar

Remarks:

RccHan™:WIST

Sex:

female

Details on test animals or test system and environmental conditions:

Female Wistar (RccHan™:WIST) strain rats were supplied by Envigo RMS (UK) Limited, Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non pregnant. After an acclimatization period of at least 5 days the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on a cage card. At the start of the study the animals were 8 to 12 weeks of age. The body weight variation did not exceed ±20% of the mean body weight at the start of treatment.

Animal Care and Husbandry
The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analyzed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
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:

other: Arachis Oil was used as the vehicle for the 300 mg/kg dose level. For the 2000 mg/kg dose level the test material was used undiluted (no vehicle)

Details on oral exposure:

In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose.
A single female animal was treated as follows:
Dose Level (mg/kg) Concentration (mg/mL) Dose Volume (mL/kg)
300 30 10

In the absence of toxicity at a dose level of 300 mg/kg, an additional female animal was treated as follows:
Dose Level (mg/kg) Specific Gravity Dose Volume (mL/kg)
2000 0.966 2.08

In the absence of toxicity at a dose level of 2000 mg/kg, an additional group of 4 female animals was treated as follows:
Dose Level (mg/kg) Specific Gravity Dose Volume (mL/kg)
2000 0.966 2.08

A total of five animals were therefore treated at a dose level of 2000 mg/kg in the study.
All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted body weight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each dose group to confirm the survival of the previously dosed animals.

Doses:

300 and 2000 Mg/kg bw

No. of animals per sex per dose:

300 mg/kg : 1 female
2000 mg/kg : 5 females

Control animals:

no

Details on study design:

Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for up to 14 days. Morbidity and mortality checks were made twice daily, early and late during normal working days, and once daily at weekends and public holidays.
Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death, except for animals 3-0 to 3-2 which were not weighed Day 14 due to a technician error.
At the end of the observation period the surviving 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.

Data Evaluation
The test item was classified according to Annex 3 of the OECD Guidelines for Testing of Chemicals No. 420 "Acute Oral Toxicity - Fixed Dose Method" (adopted 17 December 2001).
Evaluation of data included identification of the number of animals that died during the study (or that were killed for humane reasons), and determination of the nature, severity, onset and duration of the toxic effects. If possible, the signs of evident toxicity were described. Evident toxicity refers to the toxic effects of sufficient severity that administration of the next higher dose level could result in development of severe signs of toxicity and probable mortality. Effects on body weights and abnormalities noted at necropsy were also identified.
Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made.
The results were also evaluated according to Regulation (EC) No. 1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures

Results and discussion

Preliminary study:

300 mg/kg bw: No adverse effects

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No mortality at 300 mg/kg bw.
One female was found dead 24 h after dosing at 2000 mg/kg bw

Clinical signs:

other: 300 mg/kg bw: No signs of systemic toxicity were noted during the observation period. 2000 mg/kg bw: No signs of systemic toxicity were noted in the initial treated animal during the observation period. Hunched posture was noted in three animals 1 day af

Gross pathology:

300 mg/kg bw: No abnormalities were noted at necropsy.

2000 mg/kg bw: Abnormalities noted at necropsy of the animal that died during the study were hemorrhagic lungs, dark liver and dark kidneys. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Other findings:

Dissiminated (delete when editing) Fill

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral toxicity test showed an LD50 of 2000 mg/kg bw

Executive summary:

Acute oral toxicity: In this study, 5 female rats were administered the substance at dose levels of 300 and 2000 mg/kg bw. At 300 mg/kg bw the rat showed no mortality, clinical signs, body weight changes or abnormalities at necropsy.

At 2000 mg/kg bw one rat showed died, in the surviving 4 females rats there were no clinical signs, no unexpected body weight changes or abnormalities seen at necropsy.

The acute oral LD50 for the substance in female rats was determined to be 2000 mg/kg bw.