Poly[oxy(methyl-1,2-ethanediyl)], α-hydro-ω-hydroxy-, ether with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol (3:1), 2-propenoate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

key study

Study period:

From May 17, 2010 to June 09, 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

Justification for type of information:

Refer to the section 13 for details on the read across justification. The short-term toxicity to aquatic invertebrates study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Remarks:

LC-MS/MS

Details on sampling:

- Concentrations: All concentration levels and the control were analytically verified in the fresh media (0 h) and in the old media (48 h).
- Sampling method: For the old media, separate replicates without test organisms were prepared and incubated under test conditions. The method was validated according to SANCO 3029/99 rev.4 (2000-07-11).
- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until start of analysis if necessary.

Vehicle:

no

Details on test solutions:

Preparation and application of the test solution (especially for difficult test substances)
- Method: A stock solution of 100 mg/L was freshly prepared with dilution water. The stock solution was shaken with 20 rpm for 24 h (rotating shaker 3040, GFL) at room temperature.
- Eluate: Dilution water
- Differential loading: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
- Controls: Dilution water without test substance tested under the same conditions as the test groups.

Test organisms (species):

Daphnia magna

Details on test organisms:

Test organism:
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu), further own breeding
- Age at study initiation (mean and range, SD): 2 to 24 h
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 20 +/- 2 °C, in an incubator, 16 h illumination, illumination strength max. 20 µExm-2 x s-1
- Feeding during test: No feeding
Acclimatization:
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding
- Health during acclimation (any mortality observed): Healthy, no mortality observed

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Dilution water at test start: 267 (mg CaCO3/L)

Test temperature:

20.1 - 21.6 °C

pH:

- Water Quality Parameters at Test Start (0 h, Fresh Media) (measured in one replicate per concentration and control)
Nominal - test substance - concentration
mg/L pH-Value
100 7.65
50.0 7.64
25.0 7.64
12.5 7.66
6.25 7.55
Control 7.92
- Water Quality Parameters at Test End (48 h, Old Media) (measured in all replicates)
Nominal - test substance - concentration
mg/L pH-Values
Replicates
1 2 3 4
100 7.56 7.57 7.54 7.55
50.0 7.58 7.20 7.34 7.48
25.0 7.60 7.58 7.59 7.60
12.5 7.59 7.38 7.57 7.59
6.25 7.57 7.56 7.57 7.60
Control 7.64 7.65 7.62 7.53

Dissolved oxygen:

- Water Quality Parameters at Test Start (0 h, Fresh Media) (measured in one replicate per concentration and control)
Nominal - test substance - concentration
mg/L O2-Concentration (mg/L)
100 8.65
50.0 8.65
25.0 8.66
12.5 8.68
6.25 8.68
Control 8.74
- Water Quality Parameters at Test End (48 h, Old Media) (measured in all replicates)
Nominal - test substance - concentration
mg/L O2 -Concentration (mg/L)
Replicates
1 2 3 4
100 8.11 8.17 8.18 8.16
50.0 8.22 8.15 8.22 8.18
25.0 8.39 8.37 8.38 8.39
12.5 8.46 8.41 8.42 8.42
6.25 8.47 8.46 8.49 8.49
Control 8.62 8.53 8.54 8.52

Salinity:

Freshwater

Conductivity:

-

Nominal and measured concentrations:

Please refer to "Any other information on materials and methods"

Details on test conditions:

Test system:
- Test vessel: Glass beakers, 50 mL
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: Glass beakers (4 cm ID x 7 cm H), 50 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): One application at test start, static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None
Test medium and water parameters:
- Source/preparation of dilution water: Acc. to Directive 92/69/EEC Method C.2, Annex 1
- Conductivity: Dilution water at test start: 640 µS/cm
- Culture medium different from test medium: Elendt M4 acc. to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: At test start and test end
Other test conditions:
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE x m-2 x s-1

Effect parameters measured (with observation intervals if applicable) :
Immobility was assessed in all treatment levels after 24 h and 48 h. An animal was considered to be immobile, if it was not able to swim during a 15 second observation period following gentle agitation of the test vessel.

Test concentrations:
- Spacing factor for test concentrations: 2
- Range finding study
Immobilization Rates of the Preliminary Test in (%) (n = 20, divided into 2 replicates with 10 daphnids each)
Nominal - test substance - concentration
(mg/L) Immobilization (%)
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 60 60 60 100 100 100
10 0 0 0 0 0 0
Control 0 0 0 0 0 0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

70.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Interval: 61.1-81.8 mg/L

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LC10

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: None
- Mortality of control: 0%
- Other adverse effects in control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50/LC50: 1.70 (CI 1.51 - 1.97) mg/L

Reported statistics and error estimates:

The EC10- and the EC50-values were calculated by sigmoidal dose -response regression. The confidence intervals for the EC50-values were calculated from the best-fit values, the standard error and the t-distribution with the software GraphPad prism.

Table 1. Immobilization rates of the definitive test after 24 and 48 h of exposure

                 (n = 20, divided into 4 replicates with 5 daphnids each)

Nominal Test Substance Concentration	Immobilization [%]
	24 h					48 h
	Replicates					Replicates
[mg/L]	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
100	40	60	60	60	55	60	100	100	100	90
50.0	0	0	0	0	0	0	40	0	0	10
25.0	0	0	0	0	0	0	0	0	0	0
12.5	0	0	0	0	0	0	0	0	0	0
6.25	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

MV: Mean Value

The test solutions of all concentration levels were visually clear throughout exposure. The recoveries in the fresh (0 h) and old media (48 h) were in the range of 96 to 107 % of the nominal values, demonstrating that concentrations of the test substance were successfully maintained for the duration of the test. All effect values given are based on the nominal concentrations of the test substance.

Validity criteria fulfilled:

yes

Conclusions:

Based on the results of the read across study, the 48h EC50 and NOEC (Daphnia magna) were determined to be 70.7 and 50 mg/L (nominal), respectively.

Executive summary:

A study was conducted to determine the short-term toxicity to aquatic invertebrates of the read across substance, TMP(EO)TA according to OECD Guideline 202 and EU Method C.2, in compliance with GLP. The study was conducted under static conditions with 5 concentration levels of the test substance ranging from 6.25 to 100 mg/L over a period of 48 h. 20 test organisms were exposed to each test concentration and to the control. All concentration levels and the control were analysed by LC-MS/MS after 0 h and 48 h. The recoveries in the fresh (0 h) and old media (48 h) were in the range of 96 to 107% of the nominal values, demonstrating that concentrations of the test substances were successfully maintained for the duration of the test. Water quality parameters of pH-value and dissolved oxygen concentration, measured at 0 and 48 h, were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled. Immobilization was recorded in groups after 24 h and 48 h at the concentrations of 100 mg/L and only in one replicate after 48 h at the concentration of 50 mg/L (Noack, 2010). Based on the results of the read across study, the 48h EC50 and NOEC (Daphnia magna) of the test substance can be considered to be at 70.7 and 50 mg/L (nominal), respectively.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 17, 2010 to June 09, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

- Name of test material (as cited in study report): Photomer 4149 F
- Molecular formula: C21H32O9
- Molecular weight: 428.48 g/mol
- Physical state: Liquid, clear light yellowish
- Analytical purity: 100 %
- Purity test date: 2010-03-19
- Lot/batch No.: S700040031
- Stability under test conditions: Not specified
- Storage condition of test material: 6 +/- 2 °C, protected from moisture and light

Analytical monitoring:

yes

Remarks:

LC-MS/MS

Details on sampling:

- Concentrations: All concentration levels and the control were analytically verified in the fresh media (0 h) and in the old media (48 h).
- Sampling method: For the old media, separate replicates without test organisms were prepared and incubated under test conditions. The method was validated according to SANCO 3029/99 rev.4 (2000-07-11).
- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until start of analysis if necessary.

Vehicle:

no

Details on test solutions:

Preparation and application of the test solution (especially for difficult test substances)
- Method: A stock solution of 100 mg/L was freshly prepared with dilution water. The stock solution was shaken with 20 rpm for 24 h (rotating shaker 3040, GFL) at room temperature.
- Eluate: Dilution water
- Differential loading: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
- Controls: Dilution water without test substance tested under the same conditions as the test groups.

Test organisms (species):

Daphnia magna

Details on test organisms:

Test organism:
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu), further own breeding
- Age at study initiation (mean and range, SD): 2 to 24 h
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 20 +/- 2 °C, in an incubator, 16 h illumination, illumination strength max. 20 µExm-2 x s-1
- Feeding during test: No feeding
Acclimatization:
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding
- Health during acclimation (any mortality observed): Healthy, no mortality observed

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Dilution water at test start: 267 (mg CaCO3/L)

Test temperature:

20.1 - 21.6 °C

pH:

- Water Quality Parameters at Test Start (0 h, Fresh Media) (measured in one replicate per concentration and control)
Nominal - test substance - concentration
mg/L pH-Value
100 7.65
50.0 7.64
25.0 7.64
12.5 7.66
6.25 7.55
Control 7.92
- Water Quality Parameters at Test End (48 h, Old Media) (measured in all replicates)
Nominal - test substance - concentration
mg/L pH-Values
Replicates
1 2 3 4
100 7.56 7.57 7.54 7.55
50.0 7.58 7.20 7.34 7.48
25.0 7.60 7.58 7.59 7.60
12.5 7.59 7.38 7.57 7.59
6.25 7.57 7.56 7.57 7.60
Control 7.64 7.65 7.62 7.53

Dissolved oxygen:

- Water Quality Parameters at Test Start (0 h, Fresh Media) (measured in one replicate per concentration and control)
Nominal - test substance - concentration
mg/L O2-Concentration (mg/L)
100 8.65
50.0 8.65
25.0 8.66
12.5 8.68
6.25 8.68
Control 8.74
- Water Quality Parameters at Test End (48 h, Old Media) (measured in all replicates)
Nominal - test substance - concentration
mg/L O2 -Concentration (mg/L)
Replicates
1 2 3 4
100 8.11 8.17 8.18 8.16
50.0 8.22 8.15 8.22 8.18
25.0 8.39 8.37 8.38 8.39
12.5 8.46 8.41 8.42 8.42
6.25 8.47 8.46 8.49 8.49
Control 8.62 8.53 8.54 8.52

Salinity:

Freshwater

Conductivity:

-

Nominal and measured concentrations:

Please refer to "Any other information on materials and methods"

Details on test conditions:

Test system:
- Test vessel: Glass beakers, 50 mL
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: Glass beakers (4 cm ID x 7 cm H), 50 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): One application at test start, static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None
Test medium and water parameters:
- Source/preparation of dilution water: Acc. to Directive 92/69/EEC Method C.2, Annex 1
- Conductivity: Dilution water at test start: 640 µS/cm
- Culture medium different from test medium: Elendt M4 acc. to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: At test start and test end
Other test conditions:
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE x m-2 x s-1

Effect parameters measured (with observation intervals if applicable) :
Immobility was assessed in all treatment levels after 24 h and 48 h. An animal was considered to be immobile, if it was not able to swim during a 15 second observation period following gentle agitation of the test vessel.

Test concentrations:
- Spacing factor for test concentrations: 2
- Range finding study
Immobilization Rates of the Preliminary Test in (%) (n = 20, divided into 2 replicates with 10 daphnids each)
Nominal - test substance - concentration
(mg/L) Immobilization (%)
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 60 60 60 100 100 100
10 0 0 0 0 0 0
Control 0 0 0 0 0 0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

70.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Interval: 61.1-81.8 mg/L

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LC10

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: None
- Mortality of control: 0%
- Other adverse effects in control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50/LC50: 1.70 (CI 1.51 - 1.97) mg/L

Reported statistics and error estimates:

The EC10- and the EC50-values were calculated by sigmoidal dose -response regression. The confidence intervals for the EC50-values were calculated from the best-fit values, the standard error and the t-distribution with the software GraphPad prism.

Table 1. Immobilization rates of the definitive test after 24 and 48 h of exposure

                 (n = 20, divided into 4 replicates with 5 daphnids each)

Nominal Test Substance Concentration	Immobilization [%]
	24 h					48 h
	Replicates					Replicates
[mg/L]	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
100	40	60	60	60	55	60	100	100	100	90
50.0	0	0	0	0	0	0	40	0	0	10
25.0	0	0	0	0	0	0	0	0	0	0
12.5	0	0	0	0	0	0	0	0	0	0
6.25	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

MV: Mean Value

The test solutions of all concentration levels were visually clear throughout exposure. The recoveries in the fresh (0 h) and old media (48 h) were in the range of 96 to 107 % of the nominal values, demonstrating that concentrations of the test substance were successfully maintained for the duration of the test. All effect values given are based on the nominal concentrations of the test substance.

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditons, the 48h EC50 and NOEC (Daphnia magna) were determined to be 70.7 and 50 mg/L (nominal), respectively.

Executive summary:

A study was conducted to determine the short-term toxicity to aquatic invertebrates of the test substance according to OECD Guideline 202 and EU Method C.2, in compliance with GLP. The study was conducted under static conditions with 5 concentration levels of the test substance ranging from 6.25 to 100 mg/L over a period of 48 h. 20 test organisms were exposed to each test concentration and to the control. All concentration levels and the control were analysed by LC-MS/MS after 0 h and 48 h. The recoveries in the fresh (0 h) and old media (48 h) were in the range of 96 to 107% of the nominal values, demonstrating that concentrations of the test substances were successfully maintained for the duration of the test. Water quality parameters of pH-value and dissolved oxygen concentration, measured at 0 and 48 h, were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled. Immobilization was recorded in groups after 24 h and 48 h at the concentrations of 100 mg/L and only in one replicate after 48 h at the concentration of 50 mg/L. Under the study conditions, the 48h EC50 and NOEC (Daphnia magna) were determined to be 70.7 and 50 mg/L (nominal), respectively (Noack, 2010).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

70.7 mg/L

Additional information

A study was conducted to determine the short-term toxicity to aquatic invertebrates of the read across substance, TMP(EO)TA according to OECD Guideline 202 and EU Method C.2, in compliance with GLP. The study was conducted under static conditions with 5 concentration levels of the test substance ranging from 6.25 to 100 mg/L over a period of 48 h. 20 test organisms were exposed to each test concentration and to the control. All concentration levels and the control were analysed by LC-MS/MS after 0 h and 48 h. The recoveries in the fresh (0 h) and old media (48 h) were in the range of 96 to 107% of the nominal values, demonstrating that concentrations of the test substances were successfully maintained for the duration of the test. Water quality parameters of pH-value and dissolved oxygen concentration, measured at 0 and 48 h, were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled. Immobilization was recorded in groups after 24 h and 48 h at the concentrations of 100 mg/L and only in one replicate after 48 h at the concentration of 50 mg/L (Noack, 2010). Based on the results of the read across study, the 48h EC50 and NOEC (Daphnia magna) of the test substance can be considered to be at 70.7 and 50 mg/L (nominal), respectively.