Alcohols, C12-14, ethoxylated

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:

experimental study

Adequacy of study:

key study

Study period:

01 Mar - 13 Mar 2021

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)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations
- Sampling method: sampling at 0 h and 24 (fresh solutions) and after 48 h (aged solutions)
- Sample storage conditions before analysis: The samples were mixed with 10 mL acetonitrile and stored deep-frozen (≤ - 18 °C) until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution with a nominal concentration of 10 mg/L was prepared by adding the necessary amount of test item to a volumetric flask. Test medium was added and the solution was shaken and stirred for 60 minutes at 1000 rpm. The solution was observed to be clear and transparent; foam was observed. Test item concentrations were prepared by dilution with test medium.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: Clone V
- Age at study initiation (mean and range, SD): < 24 h
- Method of breeding: Daphnia magna was bred as single culture (1 daphnid per 100 mL) in Elendt M4 medium. pH-value: 6.0 – 9.0, dissolved oxygen: > 60 % saturation, total hardness 140 - 250 mg/L (as CaCO3), fed with single cell green algae (Desmodesmus subspicatus, formerly Scenedesmus subspicatus) at least three times a week.
- Source: bred at testing faciltiy, originally obtained from the Federal Environment Agency in Berlin/Germany.
- Feeding during test: none

ACCLIMATION
- Acclimation period: none

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

250 mg/L CaCO3

Test temperature:

19.6 – 20.4 °C

pH:

7.73 – 7.95

Dissolved oxygen:

≥ 8.03 mg/L

Nominal and measured concentrations:

Nominal test substance concentrations: 0.0625, 0.125, 0.250, 0.500, 1.00 mg/L
Time weighted mean concentrations: 0.0274, 0.0455, 0.0689, 0.116, 0.201 mg/L (based on 2 representative constituents)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers (nominal: 100 mL)
- Type (delete if not applicable): covered by glass plate
- Volume of solution: ≥ 50 mL
- Aeration: none
- Renewal rate of test solution (frequency): after 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Loading rate: ≥ 10 mL / animal

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium
- Culture medium different from test medium: no
- Intervals of water quality measurement: Temperature, pH-value and oxygen concentration were measured after 0, 24 hours (aged and fresh) and 48 hours

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light and 8 hours darkness
- Light intensity: 1181 ± 104 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mobility and behavioural changes 24 and 48 h

RANGE-FINDING STUDY
- Test concentrations: 0.010, 0.10, 1.00, 10.0 mg/L
- Results used to determine the conditions for the definitive study: EC50 (48 h) >0.10 mg/L < 1 mg/L

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.125 mg/L

95% CI:

>= 0.11 - <= 0.142

Nominal / measured:

meas. (TWA)

Conc. based on:

other: based on two representative constituents

Remarks:

C12EO2 and C12EO3

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.045 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

other: based on two representative constituents

Remarks:

C12EO2 and C12EO3

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: After 24 hours 20% immobile daphnids were observed at a nominal test item concentration of 0.500 mg/L. 100% immobilisation was observed at a nominal test item concentration of 1.00 mg/L. After 48 hours 20% immobile daphnids were observed at nominal test item concentrations of 0.250 and 0.500 mg/L.
- Mortality of control: none
- Other adverse effects control: After 24 hours the mobile daphnids in the nominal test item concentration of 0.500 mg/L were sluggish. On immobile daphnids at nominal test item concentration 1.00 mg/L particles were observed. After 48 hours the mobile daphnids in the nominal test item concentrations of 0.250 and 0.500 mg/L were sluggish. On some immobile daphnids at nominal test item concentration 0.500 mg/L particles were observed.
- Immobilisation of control: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The appearance of the test solutions was assessed daily and found clear and transparent from 0.0625 to 1.0 mg/L at each assessment

Results with reference substance (positive control):

EC50 (24 h) of the reference item potassium dichromate: 0.6 - 2.1 mg/L (test started on 23 Feb 2021).

Reported statistics and error estimates:

The NOEC and LOEC were determined by using a multiple comparison method (Step-down Cochran-Armitage test procedure). The value for EC50 after 24 h was determined by Weibull analysis using linear max. likelihood regression. The EC50-value after 48 h was determined following the trimmed Spearman-Karber procedure.

Table 1: Validity criteria for OECD 202 (2004)

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	0%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	≥ 8.03 mg/L	yes

Table 2:    Results of the test, 48 h values

	Nominal test item concentration [mg/L]
	Control	0.0625	0.125	0.250	0.500	1.00
	Immobilised daphnids after 48 h
Replicate 1	0	0	0	1	2	5
Replicate 2	0	0	0	1	1	5
Replicate 3	0	0	0	1	1	5
Replicate 4	0	0	0	1	0	5
Σ	0	0	0	4	4	20
%	0	0	0	20	20	100

 

Validity criteria fulfilled:

yes

Remarks:

Please see table 1 at "any other information on results incl. tables"

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

13 - 15 FEB 2008

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

adopted 2004

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: An acetonic stock solution was prepared. The respective amounts of this stock solution were directly applied to the vessels and the acetone was evaporated. Subsequently test medium was added and the test solutions were moderately stirred overnight followed by filtration (glassfibre filter). The resulting water accomodated fractions wre used in the test.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: waterflea
- Length at study initiation (length definition, mean, range and SD): < 2 mm
- Method of breeding: parental and young Daphnia were held in 40 L glass aquaria at 20 +/- 2 °C.


ACCLIMATION
- Acclimation period: 4 h
- Type and amount of food: 15 g Sera micropan (Sera GmbH, D-52525 Heinsberg) homogenized in 1 L deionized water
- Feeding frequency: once a day, except weekends

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 ± 0.5 °C

pH:

6.7 - 7.9

Dissolved oxygen:

5.9 - 6.4 mg/L

Nominal and measured concentrations:

Nominal test substance concentrations: 1.02, 2.56, 6.40, 16.0, 40.0 and 100 mg/L (WAF)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass flasks (100 ml)
- Fill volume: 50 ml
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water
- Intervals of water quality measurement: at test start and test end

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light, 8 h darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation after 24 and 48 h

RANGE-FINDING STUDY
- Test concentrations: 1, 10 an d100 mg/L
- Results used to determine the conditions for the definitive study: 65% immobile Daphnia after 48 h at 10 mg/L

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

2.49 mg/L

95% CI:

>= 1.74 - <= 3.47

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.838 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Table 1: Validity criteria for OECD 202

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	5 -10%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	5.9 - 6.4 mg/L	yes

Table 2 Results

Nominal concentration [mg/L]	Replicate	Test period (h)
		24 h	48 h
		Immobility [%]	Immobility [%]
0	A B C	5 0 5	5 10 5
1.02	A B	0 0	5 0
2.56	A B	60 45	60 65
6.40	A B	90 75	95 80
16.0	A B	85 90	100 95
40.0	A B	95 100	100 95
100	A B	100 95	100 100

Validity criteria fulfilled:

yes

Remarks:

Please see table 1 at "any other information on results incl. tables"

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

22 - 24 JUN 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: EG-Guideline 92/69/EWG

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution was prepared by diluting 20 mg/L in demineralised water.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: waterflea
-- Strain/clone: clone 5
- Source: in-house culture
- Age at study initiation (mean and range, SD): < 24 h
- Feeding during test: none


ACCLIMATION
- Type and amount of food: Scenedesmus subspicatus ad libitum
- Feeding frequency: daily

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 +/- 1 °C

pH:

7.4 - 7.5

Dissolved oxygen:

7.1 - 8.4 mg/L

Nominal and measured concentrations:

nominal: 0.1, 0.2, 0.4, 0.8, 1.6 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:glass beaker
- Fill volume: 10 mL test solution
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: one test organism per 2 mL test solution


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: CaCl2 x 2 H2O: 294 mgL; MgSO4 x 7 H2O: 123 mg/L, NaHCO3: 63 mg/L and KCl: 5.5 mg/L
- Ca/mg ratio: 4 : 1
- Intervals of water quality measurement: test start and test end


OTHER TEST CONDITIONS
- Photoperiod: continous darkness


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mobility after 24 and 48 h

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.53 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

0.61 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

The reference substance was tested in a separate test (1993-04-20)
- Mortality: 0.9 mg/L - 30%; 1.9 mg/L - 100%

Table 1: Validity criteria for OECD 202

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	0%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	6.6 mg/L	yes

Table 2: Immobilisation

Nominal concentration [mg/L]	Test period (h)
	24 h	48 h
	Immobility [%]	Immobility [%]
Control	0	0
0.1	0	0
0.2	0	0
0.4	10	20
0.8	75	95
1.6	100	100

Validity criteria fulfilled:

not specified

Remarks:

Please see table 1 at "any other information on results incl. tables"

Conclusions:

The 48-hour acute toxicity of alcohols, C12-14, ethoxylated (2 EO) to Daphnia magna, was determined in accordance with EU Guideline 92/69/EWG. The 48-hour EC50 value was determined to be 0.53 mg/L. The test is considered valid.

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

09 - 11 DEC 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 25 ml of each of the four stock solutions were mixed and made up to one litre with distilled water.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Age at study initiation (mean and range, SD): 6-24 h
- Source: Istituto Ricera sulle Acque, Brugherio, Italy
- Feeding during test : no

ACCLIMATION
- Acclimation period:
- Acclimation conditions (same as test or not):
- Type and amount of food: green algae (Selenastrum capricornutum) and yeast (Saccharomyces cerevisae)
- Feeding frequency: daily

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

180 mg CaCO3/L

Test temperature:

21°C

pH:

7.0

Dissolved oxygen:

6.6 mg/L

Nominal and measured concentrations:

Nominal test substance concentration: 2 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers (100 ml)
- Fill volume: 25 ml
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: filtered tap water

OTHER TEST CONDITIONS
- Photoperiod: 12 h light/12 h darkness
- Light intensity: fluorescent light

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mobility after2, 4, 8, 24 and 48 h.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Table 1: Validity criteria for OECD 202

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	0%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	6.6 mg/L	yes

Table 2: cummulative immobilisation

	Observation time [h]
	2	4	8	24	48
Control	0	0	0	0	0
2 mg/L	0	0	0	0	0

Validity criteria fulfilled:

yes

Remarks:

Please see table 1 at "any other information on results incl. tables"

Description of key information

EC50 (48 h): 0.125 mg/L (TWA; OECD 202)

Additional information

The key study, investigating the acute toxicity of Alcohols, C12-14, ethoxylated to Daphnia magna, was conducted according to OECD guideline 202. The Daphnids were exposed to nominal test substance concentrations of 0.0625, 0.125, 0.250, 0.500 and 1.00 mg/L for 48 hours. The test solutions were renewed every 24 h. the test concentrations were analytically verified by HPLC. Based on two representative constituents time weighted mean concentrations of 0.0274, 0.0455, 0.0689, 0.166, and 0.201 mg/L were determined. An EC50 (48 h) of 0.125 mg/L (TWA) was determined.

Three supporting studies are available. A static test following EG-Guideline 92/69/EWG determined an EC50 (48 h) of 0.53 mg/L (nominal). A static test conducted according to OECD guideline 202 determined an EC50 (48 h) of 2.49 mg/L (nominal). A second supporting study following OECD guideline 202 determined an EC50(48 h) > 2 mg/L (nominal) in a static limit test.