Alcohols, C12-13, branched and linear, 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:

16 Nov - 08 Dec 2020

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 20 mg/L was prepared by adding the necessary amount of test item to a volumetric flask. Test medium was added and the solution was homogenised by shaking. The solution was observed to be slightly turbid and foam was observed. Test item concentrations were prepared by dilution with test medium. At the highest test concentrations foam was observed, all other solutions were observed to be clear and transparent.

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:

19.3 – 19.8 °C

pH:

7.67 – 8.03

Dissolved oxygen:

≥ 7.90 mg/L

Nominal and measured concentrations:

Nominal test substance concentrations: 0.0625, 0.125, 0.250, 0.500 and 1.00 mg/L
Time weighted mean concentrations: 0.0358, 0.0851, 0.176, 0.323 and 0.691 mg/L (based on 3 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: 1428 lux (mean)

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.:

0.238 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

other: based on three representative constituents

Remarks:

C12EO2, C12EO3, C12EO6

Basis for effect:

mobility

Remarks on result:

other: 95% confidence limits could not be determined

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.176 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

other: based on three representative constituents

Remarks:

C12EO2, C12EO3, C12EO6

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: After 24 hours the mobile daphnids in nominal test item concentrations of 0.500 mg/L appeared to be sluggish.
- Mortality of control: no
- Other adverse effects control: no
- Immobilisation of control: no
The appearance of the test solutions was assessed daily and found clear and transparent. At the nominal test item concentration of 1.00 mg/L foam was observed at 0 and 24 hours.

Results with reference substance (positive control):

EC50 (24 h) of the reference item potassium dichromate: 0.6 - 2.1 mg/L (test started on 17 Nov 2020).

Reported statistics and error estimates:

The NOEC and LOEC were determined by using a multiple comparison method (Step-down Cochran-Armitage test procedure after 24 and 48 h). The EC50-value after 24 h was determined by Weibull analysis using linear max. likelihood regression. The value for EC50 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.	5%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	≥ 7.90 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	0	5	5
Replicate 2	0	0	0	0	5	5
Replicate 3	1	0	0	0	5	5
Replicate 4	0	0	0	0	5	5
Σ	1	0	0	0	20	20
%	5	0	0	0	100	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:

12 SEP - 16 SEP 1994

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)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name:
- Strain/clone:
- Justification for species other than prescribed by test guideline:
- Age at study initiation (mean and range, SD):
- Weight at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD):
- Stage and instar at study initiation:
- Valve height at study initiation, for shell deposition study (mean and range, SD):
- Peripheral shell growth removed prior to test initiation:
- Method of breeding:
- Source: bred at testing facility
- Age of parental stock (mean and range, SD):
- Feeding during test
- Food type:
- Amount:
- Frequency:

ACCLIMATION
- Acclimation period:
- Acclimation conditions (same as test or not):
- Type and amount of food:
- Feeding frequency:
- Health during acclimation (any mortality observed):

QUARANTINE (wild caught)
- Duration:
- Health/mortality:

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS:

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20°C

pH:

7.53 - 8.04

Dissolved oxygen:

7.9 - 8.4 mg/L

Nominal and measured concentrations:

Nominal test substance concentrations: 0.1, 0.18, 0.32, 0.56 and 1 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers (15 ml)
- Fill volume: 10 ml
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water
- Intervals of water quality measurement: at 0 and 48 h

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.6 mg/L

95% CI:

>= 0.6 - <= 0.7

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

0.8 mg/L

95% CI:

>= 0.7 - <= 0.9

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: no

Concentration [mg/L]	Test period (h)
	24 h	48 h
	Immobility [%]	Immobility [%]
0.1	0	0
0.18	0	0
0.32	5	5
0.56	15	25
1.0	75	95

Description of key information

EC50 (48 h): 0.238 mg/L (TWA)

Additional information

One key study investigating the acute toxicity Alcohols, C12-13, branched and linear, ethoxylated to aquatic invertebrates is available. The GLP guideline study was conducted under semi-static conditions according to OECD 202. The test organism, Daphnia magna, was exposed to nominal test item concentrations of 0.0625, 0.125, 0.250, 0.500 and 1.00 mg/L. Test solutions were renewed every 24 hours.

The test concentrations were analytically monitored by HPLC-MS/MS. The time weighted mean measured concentrations, based on three representative constituents of the UVCB substance, were 0.0358, 0.0851, 0.176, 0.323 and 0.691 mg/L. The determined EC50 (48 h) was 0.238 mg/L (TWA).

 

In a supporting static test according to OECD 202 conducted with Daphnia magna an EC50 (48 h) of 0.6 mg/L (nominal) was determined.