Phenethyl phenylacetate

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:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed using standard test guidelines

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

Preparation of the test Solutions :
The stock solution was prepared by dissolving 15.56 mg of the test substance in 1 liters of potable water (passed through reverse osmosis system) and kept in sonicator for the sonication process of 30 min. From this stock solution required test concentration were prepared for achieving the test concentrations of 0.8 mg/L, 1.68 mg/L, 3.52 mg/L, 7.40 mg/L & 15.56 mg/L, respectively.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
Body length: average 2.28 cm
Body weight: 0.115 g (average)

The fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration.
- Duration of housing: 15 days
- Photoperiod: 12 hours light and 12 hours darkness
- Oxygen content: 7.5 (mg/l)
- pH: 7.1
- Water temperature : 28 °C
- Diet: Standard brand feed, once daily

Acclimatization: Fishes, to be used in the study, are kept in the test water for seven days immediately before testing under following conditions:
- Photoperiod: 12 hours light and 12 hours dark
- Temperature range: 21 °C
- DO range:7.2 mg/l
- Feeding: Daily once upto sixth day of accimatization
- Aeration: Continuous
- Mortality (%): During last 48 hours = < 5 %

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

37.5 mg of CaCO3

Test temperature:

23°C

pH:

7.3

Dissolved oxygen:

7.4 mg/L

Nominal and measured concentrations:

0.8 mg/L, 1.68 mg/L, 3.52 mg/L, 7.40 mg/L & 15.56 mg/L

Details on test conditions:

Photoperiod :12 hours light and 12-hour darkness

Volume of water :4 liters

No. of animals per vessel :8 fishes

Test temperature :21°C

Adaptation :Test conditions corresponded to housing conditions

Test dates :Test started at : 23/05/2015
Date of experimental termination : 27/05/2015

Withdrawal of food :1 day before and during exposure

Aeration :Aeration in test vessels was provided during the experiment after 24 hours of the start of the experiment

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

7.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: highest loading at which no mortality was observed

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

11.47 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: LC50 was calculated through geometric mean based on LC0 and LC100 value.

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

15.55 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: lowest loading at which 100% mortality was observed

Sublethal observations / clinical signs:

Table 1. Mortality and symptoms

NominalConcentration (mg/l)	Number of fish		Mortality (cumulated) after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	-		-	-	-	-	-	-	-
0.8	4	4	-		-	-	-	-	-	-	-
1.68	4	4	-		-	-	-	-	-	-	-
3.52	4	4	-		-	-	-	-	-	-	-
7.40	4	4	-		-	-	-	-	-	-	-
15.55	4	4	4		4	-	-	-	-	-	-

 

Visible Symptoms: Slow movement as compared to control

 

NominalConcentration (mg/l)	Number of fish		Symptoms after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	NS4		NS4	NS4	NS4	NS4	NS4	NS4	NS4
0.8	4	4	M4		M4	M4	M4	M4	M4	M4	M4
1.68	4	4	M4		M4	M4	M4	M4	M4	M4	M4
3.52	4	4	M4		M4	M4	M4	M4	M4	M4	M4
7.40	4	4	M4		M4	M4	M4	M4	M4	M4	M4
15.55	4	4	-		-	-	-	-	-	-	-

 

Explanation of symptoms:

A: apathy

N: narcotic-like state

C: swimming near water surface

D: swimming at the bottom of bowl

M: Slow movements compared to control

NS: No abnormal symptoms

Number behind symbol for symptom= number of affected fish

 

 Table 2. pH and oxygen content:

NominalConcentration (mg/l)	Number of fish		pH readings after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	6.5		6.6	6.5	6.7	7.2	7.2	7.2	7.1
0.8	4	4	6.5		6.5	7.1	7.1	7.2	7.2	7.3	7.3
1.68	4	4	6.7		6.7	7.2	7.2	7.1	7.1	7.1	7.1
3.52	4	4	6.3		6.3	7.1	7.1	7.0	7.0	7.0	7.0
7.40	4	4	6.5		6.5	7.2	7.2	7.1	7.1	7.1	7.1
15.55	4	4	6.5		6.5	-	-	-	-	-	-

 

NominalConcentration (mg/l)	Number of fish		Oxygen content (mg/l) after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	7.1		7.1	7.1	7.0	6.9	6.9	6.9	6.8
0.8	4	4	7.1		7.0	7.1	7.0	6.9	6.9	6.8	6.8
1.68	4	4	7.1		7.0	7.2	7.1	6.8	6.7	6.7	6.7
3.52	4	4	7.1		7.0	7.1	7.0	6.9	6.9	6.8	6.8
7.40	4	4	7.1		7.0	7.2	7.1	6.8	6.7	6.7	6.7
15.55	4	4	7.1		7.1	-	-	-	-	-	-

Table 3. Test temperature

NominalConcentration (mg/l)	Number of fish		Temperature (°C) after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	21		21	21	21	21	21	21	21
0.8	4	4	21		21	21	21	21	21	21	21
1.68	4	4	21		21	21	21	21	21	21	21
3.52	4	4	21		21	21	21	21	21	21	21
7.40	4	4	21		21	21	21	21	21	21	21
15.55	4	4	21		21	-	-	-	-	-	-

Validity criteria fulfilled:

yes

Conclusions:

Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] of test material on Zebra fish (Danio rerio) was determined to be 11.47 mg/l calculated through geometric mean based on LC0 and LC100 value.

Executive summary:

Principle of this study was to determine the effect of test chemical on the mortality rate of fishes. Test conducted in accordance with OECD Guideline 203 (Fish, Acute Toxicity Test). Zebra fish (Danio rerio) was used as a test organism. The stock solution was prepared by dissolving 15.56 mg of the test substance in 1 liters of potable water (passed through reverse osmosis system) and kept in sonicator for the sonication process of 30 min. From this stock solution required test concentration were prepared for achieving the test concentrations of 0.8 mg/L, 1.68 mg/L, 3.52 mg/L, 7.40 mg/L & 15.56 mg/L, respectively and Zebra fish (Danio rerio) were exposed to these concentration for 96 hours. Test performed under the static system. Effects on the mortality rate of fishes was calculated and were observed in the interval of 24, 48, 72 and 96 hours. The fishes were moving slowly as compared to control. No mortalities were observed in the control aquaria. Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] of test material on Zebra fish (Danio rerio) was determined to be 11.47 mg/l calculated through geometric mean based on LC0 and LC100 value. Thus based on the LC50 value, it can be concluded that the test chemical was toxic to fish and can be classified in "aquatic chronic 3"category under CLP regulation.

Description of key information

Principle of this study was to determine the effect of test chemical on the mortality rate of fishes. Test conducted in accordance with OECD Guideline 203 (Fish, Acute Toxicity Test). Zebra fish (Danio rerio) was used as a test organism. The stock solution was prepared by dissolving 15.56 mg of the test substance in 1 liters of potable water (passed through reverse osmosis system) and kept in sonicator for the sonication process of 30 min. From this stock solution required test concentration were prepared for achieving the test concentrations of 0.8 mg/L, 1.68 mg/L, 3.52 mg/L, 7.40 mg/L & 15.56 mg/L, respectively and Zebra fish (Danio rerio) were exposed to these concentration for 96 hours. Test performed under the static system. Effects on the mortality rate of fishes was calculated and were observed in the interval of 24, 48, 72 and 96 hours. The fishes were moving slowly as compared to control. No mortalities were observed in the control aquaria. Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] of test material on Zebra fish (Danio rerio) was determined to be 11.47 mg/l calculated through geometric mean based on LC0 and LC100 value. Thus based on the LC50 value, it can be concluded that the test chemical was toxic to fish and can be classified in "aquatic chronic 3"category under CLP regulation.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

11.47 mg/L

Additional information

Data available for the test chemical and structurally and functionally similar read across chemicals has been reviewed to determine the toxicity of the test chemical on fishes. The studies are as mentioned below:

 

Principle of this study was to determine the effect of test chemical on the mortality rate of fishes. Test conducted in accordance with OECD Guideline 203 (Fish, Acute Toxicity Test). Zebra fish (Danio rerio) was used as a test organism. The stock solution was prepared by dissolving 15.56 mg of the test substance in 1 liters of potable water (passed through reverse osmosis system) and kept in sonicator for the sonication process of 30 min. From this stock solution required test concentration were prepared for achieving the test concentrations of 0.8 mg/L, 1.68 mg/L, 3.52 mg/L, 7.40 mg/L & 15.56 mg/L, respectively and Zebra fish (Danio rerio) were exposed to these concentration for 96 hours. Test performed under the static system. Effects on the mortality rate of fishes was calculated and were observed in the interval of 24, 48, 72 and 96 hours. The fishes were moving slowly as compared to control. No mortalities were observed in the control aquaria. Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] of test material on Zebra fish (Danio rerio) was determined to be 11.47 mg/l calculated through geometric mean based on LC0 and LC100 value. Thus based on the LC50 value, it can be concluded that the test chemical was toxic to fish and can be classified in "aquatic chronic 3"category under CLP regulation.

 

 

Above study further supported by the second study from experimental sources. This study was designed to access the toxic effects of the test compound on the fishes. Test conducted in accordance with OECD Guideline 203 (Fish, Acute Toxicity Test). Zebra fish (Danio rerio) was used as a test organism. The stock solution was prepared by dissolving 1ml of the test substance in 1 liter of potable water (passed through reverse osmosis system) with 24 hrs of continuous stirring. From this stock solution, further test concentration was prepared for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L, respectively. Aquaria containing 4 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes each. A static procedure was used for the study. Effects on the mortality rate of fishes was calculated and were observed in the interval of 24, 48, 72 and 96 hours. The median lethal concentration (LC50) value of test chemical on Danio rerio in a 96 hours study on the basis of mortality effect was determine to be >12.5 mg/L. As 100 % mortality was observed at the concentration of 25 mg/l, thus on that basis, chemical consider to be toxic and classified in aquatic chronic category 3 as per the CLP classification criteria.

 

Similar an acute toxicity test was conducted for 96 hrs for assessing the effect of test chemical on Zebra fish (Danio rerio). The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average length of 2.7 cm and average weight of 0.19 g was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 8.8 mg/l, pH 7.5, water temperature 22°C and under a photoperiod of 16:8 hr light: dark conditions, respectively. The test chemical was prepared by dissolving 5 gm of test chemical in 5 lit of potable water (passed through reverse osmosis system). After stirring, the stock solution was filtered and analytically detected & the concentration was found to be 236.34 mg/l. The remaining test solutions were prepared by dilution from the stock solution. Test chemical concentrations were analytically determined by UV-VIS spectrophotometer. Test chemical concentrations used for the study were 0, 6.25, 12.5, 50 and 100 mg/l, respectively. Total 7 fishes were exposed to test chemical in a 4 lit Polypropylene (PP) fish tank containing 4000 ml of potable water. The test vessels were placed in a room at a temperature of 21 -22°C, pH of control at 0 and 96 hr was 7.8 & 7.9 and DO of control at 0 and 96 hr was 7.9 & 4.8 mg/l and under a photoperiod of 16:8 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. Mortality in the control was 0%. The dissolved oxygen concentration remained above 60% of the air saturation value throughout the exposure period. Thus, fulfilling the validity criterion. As the concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on nominal concentration. On the basis of effect of test chemical on mortality of the test organism, the 96 hr median lethal concentration [LC50 (96 h)] for test chemical on Danio rerio (Zebra Fish) was determined to be in the range of > 25 to < 50 mg/L. Thus, test chemical was considered as toxic to aquatic fishes and hence, considered to be classified in 'aquatic chronic category 3' as per the CLP classification criteria.

 

Thus based on the above all studies and effects observations, it is concluded that the test chemical was toxic and can be consider to be classified in aquatic chronic category 3 as per the CLP classification criteria.