Phenethyl phenylacetate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

for read across substance

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

WoE report was prepared based on three toxicity study of aquatic invertebrate for the test chemical

GLP compliance:

not specified

Analytical monitoring:

yes

Remarks:

WoE 3

Details on sampling:

2. The stock solution 100 mg/l was prepared by dissolving transparent liquid in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water.

Vehicle:

no

Details on test solutions:

WoE - 2.The stock solution 100 g/L was prepared by dissolving colourless liquid in acetone. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water.


WoE - 3: The saturated test solution was prepared by dissolving 500mg of test chemical in 500ml of M7 media, and allowed for stirring for 96 hours, which was then filtered and the final saturated stock solution obtained was 162.82 mg/L, verified analytically by UV-Vis Spectrophotometer. Further, exposure concentrations of 0, 6.25, 13.12, 27.55, 57.85 and 121.48 mg/l, respectively was from the saturated test concentrations.

WoE 4: The stock solution 100 mg/l was prepared in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water.

Test organisms (species):

Daphnia magna

Details on test organisms:

WoE 2, 4:
TEST ORGANISM
- Common name: Water flea
- Strain: Straus
- Source: Own breeding at University of Chemistry and Technology, Prague
- Age at study initiation (mean and range, SD): The animals used for the test shall be less than 24 h old and should not be first brood progeny
- Feeding during test: No feeding

WoE 3:
TEST ORGANISM
- Common name: Water flea
- Strain/clone: Eggs of Daphnia magna were obtained from MicroBio tests Kleimoer 15B-9030 MARIAKERKE (GENT) BELGIUM
- Feeding during test: no
- Other: A population of parthenogenetic females of synchronized age structure has been maintained for more than 2 years in the test facility under constant temperature conditions (18 to 22°C) at a 16 : 8 hour light-dark photoperiod (illumination: < 1000 lux). The culture media (M7 medium') was partly renewed twice a week. During the maintenance of test organism, test daphnids were fed with unicellular green algae (Selenestrum capricornutum).

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

WoE 3: > 140 mg of CaCO3

Test temperature:

WoE 2,4: 20±1°C
WoE 3: 20°C

pH:

WoE 2: Test at 80 mg/l: 7.8 (didnot changed during test)
Control: 7.8 (changed to pH 7.6 during test)
Control + acetone: 7.9 change to 7.8 during test

WoE 4: Test: 7.7 (no change during test)
Control: 7.8 (Change to 7.6 during test)

Dissolved oxygen:

WoE 2: higher than 8.1 mg/L at the end of test
WoE 4: higher than 7.8 mg/L at the end of test

Nominal and measured concentrations:

WoE 2: 0, 0, 5, 10, 20, 40 and 80 mg/L
WoE 3: Nominal concentrations: 0, 6.25, 13.12, 27.55, 57.85 and 121.48 mg/l
WoE 4: 0, 2.5, 5, 10, 20, 40 mg/l nominal concentration

Details on test conditions:

WoE 2,4:
TEST SYSTEM
- Test vessel: 50 ml glass vessel
- fill volume: 25 ml
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Natural water (surface or ground water), reconstituted water or dechlorinated tap water are acceptable as culturing and dilution water if D. magna survives in it for the duration of the culturing, acclimation and testing without showing signs of stress. Waters in the range pH 6 to pH 9, with hardness between 140 mg/l and 275 mg/l (as CaCO3) are recommended.
As an example, the preparation of dilution water meeting the requirements is described below.
Dissolve known quantities of reagents in water. The dilution water prepared shall have a pH of 7.8 ± 0.5, a hardness of (225 ± 50) mg/l (expressed as CaCO3), a molar Ca + Mg ratio close to 4 + 1 and a dissolved oxygen concentration above 7 mg/l.

Prepare the solutions specified below:
- Calcium chloride solution: Dissolve 117.6 g of calcium chloride dihydrate (CaCl2.2H2O) in water (4.2) and make up to 1 l with water (4.2).
- Magnesium sulfate solution: Dissolve 49.3 g of magnesium sulfate heptahydrate (MgSO4.7H2O) in water (4.2) and make up to 1 l with water (4.2).
- Sodium bicarbonate solution: Dissolve 25.9 g of sodium bicarbonate (NaHCO3) in water (4.2) and make up to 1 l with water (4.2).
- Potassium chloride solution: Dissolve 2.3 g of potassium chloride (KCI) in water (4.2) and make up to 1 l with water (4.2).

Mixing
Mix 2.5 ml of each of the four solutions and make up to 1 l with water.
The dilution water shall be aerated until the dissolved oxygen concentration has reached saturation and the pH has stabilized. If necessary, adjust the pH to 7.8 ± 0.5 by adding sodium hydroxide (NaOH) solution or hydrochloric acid (HCI). The dilution water prepared in this way shall not be further aerated before use.

- Sodium hydroxide solution, e.g. [NaOH] : 1 mol/l.
- Hydrochloric acid, e.g. [HCl] : 1 mol/l.

Reference substance:
Dissolve 600 mg of potassium dichromate (K2Cr2O7) in water and make up to 1 l with water (4.2).

OTHER TEST CONDITIONS
- Adjustment of pH: no adjustment done
- Photoperiod: No - Darkness
- Light intensity:

CALCULATION:
EC50 was calculated using non linear regression by the software Prism 4.0

WoE 3:
TEST SYSTEM
- Test vessel: Glass beaker
- Aeration: No aeration during experiment
- Material, size, headspace, fill volume: 25 ml of glass beaker filled with 20 ml media having headspace of 5 ml
- No. of organisms per vessel: 10 daphnids

OTHER TEST CONDITIONS
- Photoperiod:16 hours light and 8 hours dark
- Light intensity: 1000 – 1500 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The criterion of immobilisation used in this study was the absence of response to physical stimulation. In addition to observations on mortality at 24, and 48 hours, type and incidence of sub-effective effects compared with control Daphnia were observed.

Reference substance (positive control):

yes

Remarks:

potassium dichromate was used as a reference substance for the study.

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

60.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: WoE 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

63.86 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: WoE 3

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

16.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI was 14.7-18.7 mg/l

Remarks:

WoE 4

Results with reference substance (positive control):

WoE 3: The 24 hr EC50 value of reference substance potassium dichromate was determined to be 0.831 mg/l.
WoE 4:- Results with reference substance valid
- EC50: 0.79 mg/L (24 hours)

Reported statistics and error estimates:

EC50 was calculated using non linear regression by the software Prism 4.0

Validity criteria fulfilled:

not specified

Conclusions:

An acute immobilisation test was conducted for 48 hrs for assessing the short term toxicity of test chemical to aquatic invertebrate. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Based on the effects on the immobilisation of Daphnia magna after the chemical exposure for 48 hours, it was observed that the EC50 value ranges from 16.6 to 63.86 mg/l. Thus based on the overall studies from various sources, it was concluded that the test chemical was toxic and classified in aquatic chronic category 3 as per the CLP classification criteria.

Executive summary:

Data available for the test chemicals has been reviewed to determine the toxicity of the test chemical on aquatic invertebrates Daphnia magna. The studies are as mentioned below:

 

Aim of this key weight of evidence study was to assess the short term toxicity of test chemical to aquatic invertebrate. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna was used a test organism. The animals used for the test was Daphnia magna, which should be less than 24 h old and should not be of first brood progeny. The stock solution 100 g/L was prepared in acetone. Test solutions of required concentration as were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 0, 5, 10, 20, 40 and 80 mg/L concentrations were used in the study. Effects on immobilisation were observed for 48 hours and conducted under the static system. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. The median effective concentration (EC50) value of the test substance on Daphnia magna was determined to be 60.7 mg/L on the basis of immobilisation effects in a 48 hour study. Based on the EC50 value, substance consider likely to be hazardous to aquatic invertebrate Daphnia magna and can be classified in aquatic chronic 3 category as per the CLP classification criteria.

 

Above study further supported by the third weight of evidence study from experimental report. An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on Daphnia magna. The test was performed in accordance to OECD guideline No. 202“Daphnia sp., Acute Immobilization Test”. The saturated test solution was prepared by dissolving 500mg of test chemical in 500ml of M7 media, and allowed for stirring for 96 hours, which was then filtered and the final saturated stock solution obtained was 162.82 mg/L, verified analytically by UV-Vis Spectrophotometer. Further, exposure concentrations of 0, 6.25, 13.12, 27.55, 57.85 and 121.48 mg/l, respectively was from the saturated test concentration. Study was performed using 10 daphnids in a static system. Total 10 Daphnids/conc. were exposed to test chemical in 25 ml beakers in a volume of 20 ml of liquid solution containing both the chemical and media. The beakers were placed in a room at a temperature of 20°C, hardness of water > 140 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions with light intensity 1000 – 1500 Lux, respectively. One control vessel was also run simultaneously during the study. The animals in control and test chemical concentrations were exposed for a period of 48 hour. Potassium dichromate was used as a reference substance for the study. The 24 hr EC50 value of reference substance was determined to be 0.831 mg/l. No Immobility were found in the control test animals and the dissolved oxygen concentration at the end of the test in the control and test vessel was ≥ 3 mg/l, thus validity criterion of the study has been fulfilled. 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 mobility of the test organism, the median effect concentration (EC50 (48 h)) value was determined to be 63.86 mg/L. Thus, based on the EC50 value, chemical was considered as toxic to aquatic invertebrates and hence, considered to be classified in 'aquatic chronic category 3' as per CLP classification criteria.

 

Similar an acute immobilisation test was conducted for 48 hrs for assessing the short term toxicity of test chemical to aquatic invertebrate. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna was used a test organism. The stock solution 100 mg/l was prepared in reconstituted water. 0, 2.5, 5, 10, 20, 40 mg/l nominal concentrations were used in the study. Effects on immobilisation were observed for 48 hours and conducted under the static system. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. Based on the immobilisation of Daphnia magna due to the exposure of test chemical for 48 hours, the EC50 value was determine to be 16.6 mg/l. EC50 value indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified in aquatic chronic 3 category as per the CLP criteria.

 

Thus based on the overall studies from various sources, it was concluded that the test chemical was toxic and classified in aquatic chronic category 3 as per the CLP classification criteria.

Description of key information

An acute immobilisation test was conducted for 48 hrs for assessing the short term toxicity of test chemical to aquatic invertebrate. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Based on the effects on the immobilisation of Daphnia magna after the chemical exposure for 48 hours, it was observed that the EC50 value ranges from 16.6 to 63.86 mg/l. Thus based on the overall studies from various sources, it was concluded that the test chemical was toxic and classified in aquatic chronic category 3 as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

60.75 mg/L

Additional information

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

 

Aim of this key weight of evidence study was to assess the short term toxicity of test chemical to aquatic invertebrate. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna was used a test organism. The animals used for the test was Daphnia magna, which should be less than 24 h old and should not be of first brood progeny. The stock solution 100 g/L was prepared in acetone. Test solutions of required concentration as were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 0, 5, 10, 20, 40 and 80 mg/L concentrations were used in the study. Effects on immobilisation were observed for 48 hours and conducted under the static system. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. The median effective concentration (EC50) value of the test substance on Daphnia magna was determined to be 60.7 mg/L on the basis of immobilisation effects in a 48 hour study. Based on the EC50 value, substance consider likely to be hazardous to aquatic invertebrate Daphnia magna and can be classified in aquatic chronic 3 category as per the CLP classification criteria.

 

Above study further supported by the third weight of evidence study from experimental report. An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on Daphnia magna. The test was performed in accordance to OECD guideline No. 202“Daphnia sp., Acute Immobilization Test”. The saturated test solution was prepared by dissolving 500mg of test chemical in 500ml of M7 media, and allowed for stirring for 96 hours, which was then filtered and the final saturated stock solution obtained was 162.82 mg/L, verified analytically by UV-Vis Spectrophotometer. Further, exposure concentrations of 0, 6.25, 13.12, 27.55, 57.85 and 121.48 mg/l, respectively was from the saturated test concentration. Study was performed using 10 daphnids in a static system. Total 10 Daphnids/conc. were exposed to test chemical in 25 ml beakers in a volume of 20 ml of liquid solution containing both the chemical and media. The beakers were placed in a room at a temperature of 20°C, hardness of water > 140 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions with light intensity 1000 – 1500 Lux, respectively. One control vessel was also run simultaneously during the study. The animals in control and test chemical concentrations were exposed for a period of 48 hour. Potassium dichromate was used as a reference substance for the study. The 24 hr EC50 value of reference substance was determined to be 0.831 mg/l. No Immobility were found in the control test animals and the dissolved oxygen concentration at the end of the test in the control and test vessel was ≥ 3 mg/l, thus validity criterion of the study has been fulfilled. 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 mobility of the test organism, the median effect concentration (EC50 (48 h)) value was determined to be 63.86 mg/L. Thus, based on the EC50 value, chemical was considered as toxic to aquatic invertebrates and hence, considered to be classified in 'aquatic chronic category 3' as per CLP classification criteria.

 

Similar an acute immobilisation test was conducted for 48 hrs for assessing the short term toxicity of test chemical to aquatic invertebrate. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna was used a test organism. The stock solution 100 mg/l was prepared in reconstituted water. 0, 2.5, 5, 10, 20, 40 mg/l nominal concentrations were used in the study. Effects on immobilisation were observed for 48 hours and conducted under the static system. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. Based on the immobilisation of Daphnia magna due to the exposure of test chemical for 48 hours, the EC50 value was determine to be 16.6 mg/l. EC50 value indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified in aquatic chronic 3 category as per the CLP criteria.

 

Thus based on the overall studies from various sources, it was concluded that the test chemical was toxic and classified in aquatic chronic category 3 as per the CLP classification criteria.