Ethyl butyrate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09-05-2017 - 13-05-2017

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental study result was performed accoridng to the OECD guidelines.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Principles of method if other than guideline:

Short term toxicity study was performed according to the standard guideline 203 for the test material .

GLP compliance:

no

Analytical monitoring:

not specified

Details on sampling:

No data available

Vehicle:

not specified

Details on test solutions:

- Method: The test substance was sparingly soluble in water. Therefore, the test solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with 24 hours continuous stirring. Concentrations decided are 6.25mg/L,12.5mg/L, 25mg/L,50mg/L,100mg/L ,respectively.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name:Zebra Fish
- Strain:Danio rerio
- Source:Koyal aquarium,Nagpur., Nagpur, MS (India)
- Age at study initiation (mean and range, SD):No data
- Length at study initiation (length definition, mean, range and SD): average 1.89 cm
- Weight at study initiation (mean and range, SD):0.1702g (average)
- Method of breeding: No data
- Feeding during test: yes
- Food type:Standard brand feed,
- Amount: No data
- Frequency: once daily

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

No data

Post exposure observation period:

No data

Hardness:

148.5 (mg of CaCO3)

Test temperature:

23.3°C

pH:

7.62

Dissolved oxygen:

7.7 mg/L

Salinity:

No data

Conductivity:

No data

Nominal and measured concentrations:

0, 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: plastic aquaria
- Type (delete if not applicable): No data
- Material, size, headspace, fill volume: plastic aquaria having the water capacity of 5liters filled with 2 liter of water containing test chemical and headspace of 3 liters.
- Aeration: Aeration in test vessels was provided 1 day before the start of experiment (Continuous)
- Type of flow-through (e.g. peristaltic or proportional diluter): No data
- Renewal rate of test solution (frequency/flow rate): No data
- No. of organisms per vessel: 8 fishes
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): No data
- Biomass loading rate: No data

TEST MEDIUM / WATER PARAMETERS: No data
- Source/preparation of dilution water:
- Total organic carbon:
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:
- Ca/mg ratio:
- Culture medium different from test medium:
- Intervals of water quality measurement:

OTHER TEST CONDITIONS
- Adjustment of pH: yes
- Photoperiod: 16 hours light and 8 hours dark
- Light intensity: No data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : pH, Dissolved oxygen, Temperature, Mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: No data
- Justification for using less concentrations than requested by guideline: No data
- Range finding study: No data
- Test concentrations: 0, 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/L
- Results used to determine the conditions for the definitive study: Mortality

Reference substance (positive control):

not specified

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No mortality

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No mortality

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No mortality

Details on results:

No abnormalities was observed

Results with reference substance (positive control):

No data

Reported statistics and error estimates:

No data

Sublethal observations / clinical signs:

Fig:Mortality and symptoms

Nominal Concentration (mg/l)	Number of fish		Mortality (cumulated) after
			3hr			6hr		24hr			48hr		72hr			96hr
			V1	V2		V1	V2	V1	V2		V1	V2	V1	V2		V1	V2
Control	4	4	-		-	-	-	-		-	-	-	-		-	-		-
6.25	4	4	-		-	-	-	-		-	-	-	-		-	-		-
12.5	4	4	-		-	-	-	-		-	-	-	-		-	-		-
25	4	4	-		-	-	-	-		-	-	-	-		-	-		-
50	4	4	-		-	-	-	-		-	-	-	-		-	-		-
100	4	4	-		-	-	-	-		-	-	-	-		-	-		-

Fig: Visible Symptoms:

Nominal Concentration (mg/l)

Number of fish

symptom (cumulated) after

3hr

6hr

24hr

48hr

72hr

96hr

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

6.25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

12.5

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

50

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

100

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

Validity criteria fulfilled:

yes

Conclusions:

Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for test material on Zebra Fish was > 100 mg/L.

Executive summary:

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for the test substance on Zebra fish. The nominal concentration selected for the experiment were 0, 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/L and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. The test substance was sparingly soluble in water. Therefore, the test solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with 24 hours continuous stirring Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. The nominal concentration selected for the experiment was 0, 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/l. and Zebra fish were exposed to this concentration for 96 hours. The lethal concentrations LC50 was determine to be >100 mg/l. After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be >100 mg/l. Based on the LC50, it can be consider that the chemical was non toxic and can be consider to be not classified as per the CLP classification criteria.

Description of key information

Short term toxicity to fish:

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for the test substance on Zebra fish.The nominal concentration selected for the experiment were 0, 6.25mg/L,12.5mg/L, 25mg/L, 50mg/L,100mg/L and Zebra Fish Danio rerio were exposed to these concentration for 96 hours.

 

The test substance was sparingly soluble in water. Therefore, the test solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with 24 hours continuous stirring Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. The nominal concentration selected for the experiment was 0, 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/l. and Zebra fish were exposed to this concentration for 96 hours. The lethal concentrations LC50 was determine to be >100 mg/l. After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be >100 mg/l . Based on the LC50, it can be consider that the chemical was non toxic and can be consider to be not classified as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The short term toxicity of fish was summarized with detailed experimental data as per OECD guideline along with another experimental data described in peer reviewed journals and handbooks for the test chemical.

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for the test substance on Zebra fish. The nominal concentration selected for the experiment were0, 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/L and Zebra Fish Danio rerio were exposed to this concentration for 96 hours.The test substance was sparingly soluble in water. Therefore, the test solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with 24 hours continuous stirring Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. The nominal concentration selected for the experiment was 0, 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/l. and Zebra fish were exposed to this concentration for 96 hours. The lethal concentrations LC50 was found to be >100 mg/l.

After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be >100 mg/l . Based on the LC50, it can be consider that the chemical was non toxic and can be consider to be not classified as per the CLP classification criteria.

 

First study was supported by the second study. Short term toxicity of test material was evaluated on fish golden orfe in static condition for 48 h The median lethal effect LC50 concentration of test material on fish after 48 h was observed to be 17 mg/l.Based on this effect concentration it can be considered that test material is toxic to fish and can be classified in the category aquatic chronic 3 as per CLP criteria .

 

Similarly in short term toxicity to fish study for test substance in Leuciscus idus melanotus was observed for 2 days. No mortality was observed at concentration of 35 mg/l. 50% mortality was observed at range concentration of 53-78 mg/l. 100% mortality was observed at range concentration of 87-131 mg/l.As available lethal concentration indicate the test substance classified in aquatic chronic 3; since the test chemical readily biodegradable in nature thus not consider for the aquatic classification as per the CLP criteria.

 

Since, varied results on different fishes were observed the experimental data conducted occording to OECD guideline is considered for the classification of test substance. The lethal concentrations LC50 was found to be >100 mg/l. Based on the LC50, it can be consider that the chemical was non toxic and can be consider to be not classified as per the CLP classification criteria.