2-Butenoic acid, 1-ethyl-2-methylpropyl ester, (2E)-

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was conducted between 27 May 2014 and 17 June 2014.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study is considered to be a reliability 1 as it has been conducted according to OECD Test Guideline 423 using the Acute Toxic Class Method and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2-methylpentan-3-yl (2E)-but-2-enoate

EC Number:

806-544-2

Cas Number:

1370711-06-0

Molecular formula:

C10H18O2

IUPAC Name:

2-methylpentan-3-yl (2E)-but-2-enoate

Test material form:

other: Liquid

Details on test material:

Identification: IFF TM 12-209
Physical state / appearance: Extremely pale yellow liquid
Storage conditions: Approximately 4 °C in the dark

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Female Wister (RccHanTM: WIST) strain rats were supplied by Harlan Laboratories UK Ltd., 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 five 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 n a cage card. At the start of the study the animals were eight to twelve weeks of age. The body weight variation did not exceed ± 20 % of the mean body weight of the initially dosed group.
The animals were housed in groups of three in suspended solid-floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately three to four hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analysed 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 (06:00 to 18:00) and twelve 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:

DMSO

Details on oral exposure:

For the purpose of the 2000 mg/kg dose level, the test item was supplied. The specific gravity was determined and used to calculate the appropriate dose volume for the required dose level. For the purpose of the 300 mg/kg dose level, the test item was freshly prepared, as required, as a solution in dimethyl sulphpoxide. Dimethyl sulphpoxide was used because the test item did not dissolve / suspend in distilled water or arachis oil BP.
The test item was formulated within 2 hours of being applied to the test system. It is assumed that the formulation was stable for this duration.
No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.

Doses:

300 and 2000 mg/kg

No. of animals per sex per dose:

3 females / dose

Control animals:

no

Details on study design:

In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose.
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 group to confirm the survival of the previously dosed animals.
The animals were observed for deaths or overt signs of toxicity 0.5, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days. Due to a technician error, the Day 10 clinical observations for the animals treated at a dose level of 300 mg/kg body weight were not performed. This deviation was considered not to affect the purpose or integrity of the study as no signs of systemic toxicity were observed pre- or post Day 10.
Individual body weights were recorded prior to dosing and seven and fourteen days after treatment.
At the end of the observation period, the 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.

Evaluation of Data
Data evaluations included the relationship, if any, between the exposure of the animal to the test item and the incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, body weight changes, mortality and any other toxicological effects.
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, relating to the Classification, labelling and Packaging of Substances and Mixtures.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths

Clinical signs:

other: Hunched posture and ataxia were noted during the day of dosing in the second group of animals treated at a dose level of 2000 mg/kg. There were no signs of systemic toxicity noted in animals treated at a dose level of 300 mg/kg and the initial group trea

Gross pathology:

No abnormalities were noted at necropsy.

Any other information on results incl. tables

Mortality Data

Dose Level mg/kg	Sex	Number of Animals Treated	Deaths During Day of Dosing (Hours)				Deaths During Period After Dosing (Days)								Deaths
			½	1	2	4	1	2	3	4	5	6	7	8-14
300	Female	3	0	0	0	0	0	0	0	0	0	0	0	0	0/3
2000	Female	3	0	0	0	0	0	0	0	0	0	0	0	0	0/3
	Female	3	0	0	0	0	0	0	0	0	0	0	0	0	0/3

 

Individual Clinical Observations - 300 mg/kg

Dose Level mg/kg	Animal Number and Sex	Effects Noted After Dosing (Hours)				Effects Noted During Period After Dosing (Days)
		½	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
300	1-0 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	*	0	0	0	0
	1-1 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	*	0	0	0	0
	1-2 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	*	0	0	0	0

0=   No signs of systemic toxicity

*=  Observation not performed due to technician error

Individual Clinical Observations - 2000 mg/kg

Dose Level mg/kg	Animal Number and Sex	Effects Noted After Dosing (Hours)				Effects Noted During Period After Dosing (Days)
		½	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2000	2-0 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-1 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-2 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-0 Female	H	H	H	HA	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-1 Female	0	H	H	HA	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-2 Female	0	H	H	HA	0	0	0	0	0	0	0	0	0	0	0	0	0	0

0= No signs of systemic toxicity                    

H = Hunched posture                        

A = Ataxia

Individual Body Weights and Body Weight Changes - 300 mg/kg

Dose Level mg/kg	Animal Number and Sex	Body Weight (g) at Day			Body Weight Gain (g) During Week
		0	7	14	1	2
300	1-0 Female	173	184	188	11	4
	1-1 Female	185	196	200	11	4
	1-2 Female	171	178	187	7	9

Individual Body Weights and Body Weight Changes - 2000 mg/kg

Dose Level mg/kg	Animal Number and Sex	Body Weight (g) at Day			Body Weight Gain (g) During Week
		0	7	14	1	2
2000	2-0 Female	174	184	204	10	20
	2-1 Female	169	176	185	7	9
	2-2 Female	180	187	196	7	9
	3-0 Female	172	203	212	31	9
	3-1 Female	169	183	196	14	13
	3-2 Female	164	180	190	16	10

 

Individual Necropsy Findings - 300 mg/kg

Dose Level mg/kg	Animal Number and Sex	Time of Death	Macroscopic Observations
300	1-0 Female	Killed Day 14	No abnormalities detected
	1-1 Female	Killed Day 14	No abnormalities detected
	1-2 Female	Killed Day 14	No abnormalities detected

 Individual Necropsy Findings - 2000 mg/kg

Dose Level mg/kg	Animal Number and Sex	Time of Death	Macroscopic Observations
2000	2-0 Female	Killed Day 14	No abnormalities detected
	2-1 Female	Killed Day 14	No abnormalities detected
	2-2 Female	Killed Day 14	No abnormalities detected
	3-0 Female	Killed Day 14	No abnormalities detected
	3-1 Female	Killed Day 14	No abnormalities detected
	3-2 Female	Killed Day 14	No abnormalities detected

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System – Category 5 > 2000 – 5000 mg/kg body weight).
The test item does not meet the criteria for classification according to Regulation (EC) No 1272 / 2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.

Executive summary:

The acute oral toxicity of the test substance, TM 12-209, was estimated to be greater than 2000 mg/kg body weight according to OECD Test Guideline 423 using the Acute Toxic Class Method.