Ethyl butyrate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

data is from experimental reports following standard procedures.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Principles of method if other than guideline:

To determine the acute oral toxicity of test chemical in Sprague Dawley rats

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl butyrate

EC Number:

203-306-4

EC Name:

Ethyl butyrate

Cas Number:

105-54-4

Molecular formula:

C6H12O2

IUPAC Name:

ethyl butanoate

Test material form:

liquid

Specific details on test material used for the study:

- Substance Type: Organic
- Physical State: Liquid

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 0001
- Expiration date of the lot/batch:July; 2021
- Manufacturing Date: July; 2016

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Test Item and prepared formulation(s) were stored at ambient temperature.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The dose of 300 mg/kg of test item was prepared by dilution of the test item in distilled water to obtain 30 mg/ml strength of solution. The formulation was prepared fresh on the day of dosing. The test item was administered in the dose volume of 10 ml/kg body weight. Actual preparation procedures are documented in the raw data. The dose of 2000 mg/kg of test item was administered undiluted as supplied by the Sponsor.

OTHER - Safety Precautions: Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females (if applicable) nulliparous and non-pregnant: [yes/no] : yes
- Age at study initiation: Healthy young adult females were used. The age at study initiation was 8 to 12 weeks old
- Weight at study initiation: Body weights at the start : Female Mean : 199.48 g (= 100 %); Minimum : 194.3 g (- 2.60 %); Maximum : 207.2 g (+ 3.87 %)
- Fasting period before study: fasted rats (approximately 16 hours or more)
- Housing:polycarbonate cages.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum):Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period:All animals were acclimatized to laboratory conditions for minimum of 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.4 to 22.0 degree centigrade
- Humidity (%): 53.1% to 58.2%.
- Air changes (per hr): ten to fifteen air changes per hour of 100% fresh air that had been passed through the HEPA filters
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: From: 31-05-2017 To: 19-06-2017

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 300 mg/kg and 2000 mg/kg
- Amount of vehicle (if gavage):10 ml per kg of body weight

MAXIMUM DOSE VOLUME APPLIED: 10 ml per kg of body weight was considered the maximum volume which could be administered to a rat.

DOSAGE PREPARATION (if unusual): The dose of 300 mg/kg of test item was prepared by dilution of the test item in distilled water to obtain 30 mg/ml strength of solution.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:The doses were selected based on the available literature of the test item.

Doses:

Dose Group I : 300 mg/kg
Dose Group II : 2000 mg/kg

No. of animals per sex per dose:

Three females were used at each step

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical Observations and General Appearance: Animals were observed for clinical signs, mortality and morbidity, until sacrifice.Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at immediately (0 to 5 minutes), 5, 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time. The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.
Body weights: Individual animal body weights were recorded, before fasting, prior to administration of the test item (fasting body weights), weekly thereafter and at termination on day 14. Weight changes were calculated and recorded.
Gross Pathology: Necropsy was performed on all animals at the end of the study period on day 15. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique.
Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.

Statistics:

Mean ± SD

Results and discussion

Preliminary study:

not specified

Effect levels

Key result

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 all the tested concentrations at all times

Clinical signs:

other: No clinical signs and signs of toxicity were observed.

Gross pathology:

Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.

Other findings:

- Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.

Any other information on results incl. tables

Table 1: Summary of Clinical Signs of Toxicity and Mortality

Sprague Dawley Rat

Sex : Female

Group I :   

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	300	No clinical signs observed	3	1,2,3	Day 0 - Day 14	0/3

 

 

 

Group I :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
II	300	No clinical signs observed	3	4,5,6	Day 0 - Day 14	0/3

 

Group II :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	2000	No clinical signs observed	3	7,8,9	Day 0 - Day 14	0/3

 

Group II :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
II	2000	No clinical signs observed	3	10,11, 12	Day 0 - Day 14	0/3

 

 

 

Table 2: Mean Body Weight and Percent Body Weight Gain (g)

Sprague Dawley Rat

Sex : Female

Group I :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	300	Mean	199.03	205.60	3.32	223.10	8.51	12.11
		± SD	3.23	1.90	2.28	2.48	0.63	2.11

 

 

Group I :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
II	300	Mean	203.43	209.37	2.91	227.20	8.52	11.68
		± SD	4.60	5.75	0.75	5.64	0.37	0.43

 

 

Group II :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	199.17	206.67	3.77	227.90	10.27	14.43
		± SD	2.14	2.81	1.00	4.19	0.67	1.78

 

 

Group II :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
II	2000	Mean	196.30	201.80	2.80	220.83	9.44	12.50
		± SD	2.17	2.61	0.20	1.40	1.13	0.94

 

Table 3: Summary of Gross Pathological Findings

Sprague Dawley Rat

Sex : Female

 

        Group I :

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	300	1 - 3	TS	No abnormality detected

 

 

        Group I :

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	300	4 - 6	TS	No abnormality detected

 

 

                    Group II :

 

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	7 - 9	TS	No abnormality detected

 

 

                    Group II :

 

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	2000	10 - 12	TS	No abnormality detected

 

                     TS = Terminal Sacrifice

 

                    

 

 

 

Applicant's summary and conclusion

Interpretation of results:

other: Not classified

Conclusions:

The acute oral LD50 value for test chemical can be considered to be >2000mg/kg. Hence, it was concluded that the acute toxicity of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.

Executive summary:

The study was designed and conducted to determine the acute oral toxicity profile of test chemical in Sprague Dawley rats. The study was performed according to OECD 423 - Acute Toxic Class Method. Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Group I Step I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Group I Step II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality after the dosing. As no mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Group II Step I). Administration of the test item at 2000 mg/kg did not result in any signs of toxicity and mortality after the dosing. As no mortality were observed at 24 hours after the dosing, additional three female animals were treated with the higher dose of  2000 mg/kg of the test item (Group II Step II). Administration of the test item at 2000 mg/kg did not result in any signs of toxicity and mortality after the dosing. No mortality or gross pathological abnormalities were observed in animals from 300 mg/kg and 2000 mg/kg dose groups. Therefore, the acute oral LD50 value for test chemical can be considered to be >2000mg/kg. Hence, it was concluded that the acute toxicity of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.