Alcohols, C12-13, branched and linear, ethoxylated, sulfates, sodium salts

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 Jul - 16 Aug 2022

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:

adopted 13 April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All concentrations including the control
- Sampling method: Samples were taken at 0 h (initial value) and after 24 h from fresh test solutions and after 24 h and 48 h from aged test solutions. Additionally, an analytical sample was taken from the stock solutions at test start and after 24 hours. 10 mL test medium sample were added with 10 mL methanol.
- Sample storage conditions before analysis: All samples were stored deep frozen (≤ - 18 °C) until they were transferred to the analytical laboratory. On the day of analysis, the samples were thawed to ambient temperature and shaken well using a vortex mixer. If necessary, the samples were further diluted with methanol/test medium (1:1, v/v) prior to analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The necessary amount of test item for preparing the stock solution was weighed into a glass vial and 20 mL test medium was added. The solution was homogenised by shaking. Lower concentrated stock solutions were prepared by dilution of appropriate solutions with test medium. In this process a stock solution for each test item concentration was produced. All stock solutions were clear and transparent, frothing was observed in all stock solutions. For the preparation of the test solutions, defined volumes of the prepared stock solutions were added to test medium. All solutions were clear and transparent, frothing was observed in three highest test solution concentrations. ≥ 50 mL of the prepared solutions were transferred to each test vessel. The preparation procedure was repeated after 24 hours.
- Controls: Test medium
- Test concentration separation factor: 2

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain/clone: Clone V
- Age at study initiation: ≤ 24 h
- Method of breeding: Daphnia magna was bred as single culture (1 daphnid per 100 mL) in Elendt M4 medium
- Source: Federal Environment Agency in Berlin/Germany
- Feeding during test: No
- Food type: Single cell green algae (Desmodesmus subspicatus)
- Frequency: 3 times a week

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

250 mg/L as CaCO3

Test temperature:

19.5 – 20.2 °C

pH:

7.86 – 8.03

Dissolved oxygen:

≥ 8.62 mg/L

Nominal and measured concentrations:

Nominal test concentrations: 23.0, 11.5, 5.75, 2.87, 1.44 and 0.00 mg/L, or 16.0, 8.00, 4.00, 2.00, 1.00 and 0.00 mg active ingredient/L C12-13 AES
TWA measured test item concentrations (substance activity considered): 16.0, 7.73, 4.09, 2.05 and 0.981 mg/L and

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass vessel (100 mL)
- Type: covered with a glass plate
- Material, size, fill volume: glass, 100 mL, ≥ 50 mL
- Volume of solution: ≥ 50 mL
- Renewal rate of test solution: after 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium
- Culture medium different from test medium: no
- Intervals of water quality measurement: 0 h fresh, 24 h aged, 24 h fresh, 48 h aged

OTHER TEST CONDITIONS
- Photoperiod: 16 h light / 8 h darkness
- Light intensity: 1166 lux (mean)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility and behavioral changes every 24 h

RANGE-FINDING STUDY
non-GLP
- Test concentrations: 0.1, 1.0, 10.0, 100.0
- Results used to determine the conditions for the definitive study: EC50 (48 h): 10 - 100 mg/L

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

7.94 mg/L

95% CI:

>= 6.74 - <= 9.35

Nominal / measured:

meas. (TWA)

Conc. based on:

other: based on six representative constituents

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: No
- Mortality of control: No
- Immobilisation of control: No
- Any observations that might cause a difference between measured and nominal values: frothing was observed in the three highest test solutions

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Dose-response test: yes
- 48 h-EC50: 0.841 mg/L (95% CI : 0.735 - 0.962)

Reported statistics and error estimates:

The statistical evaluation of the data was performed with ToxRat Professional, Version 3.3.0. ToxRat Professional. The EC50-values after 48 h was determined following the trimmed Spearman-Karber procedure.

Table 1: Nominal and calculated TWA concentrations of the test substance after 48 h:

Test item nominal	Time-weighted average (TWA) concentration [mg a.i./L]							TWA conc. test item	TWA conc. test item
[mg/L]^1)	C12EO0	C12EO1	C12EO2	C13EO0	C13EO1^2)	C13EO2^2)	Sum	[mg/L]	[mg/L]^1)
1.00	0.236	0.0464	0.0387	0.291	0.0681	0.0461	0.726	1.38	0.960
2.00	0.500	0.100	0.0846	0.571	0.136	0.0918	1.48	2.81	1.96
4.00	1.01	0.206	0.171	1.20	0.272	0.184	3.04	5.78	4.02
8.00	2.01	0.426	0.361	2.38	0.544	0.368	6.09	11.6	8.07
16.0	3.70	0.796	0.686	4.56	1.09	0.736	11.6	22.0	15.3

¹⁾ Corrected for substance activity

²⁾ Since all analytical recoveries of this constituent were within 80 – 120 % of nominal, the nominal concentrations of the constituent were used for the further calculations

 

 

Table 2: Results of the test, 48 h values

Nominal test item concentration [mg/L]^1)	Control	1.00	2.00	4.00	8.00	16.0
Time weighted average concentrations  [mg/L]^1)	Control	0.960	1.96	4.02	8.07	15.3
	Immobilised daphnids after 48 h
Replicate 1	0	0	0	1	2	5
Replicate 2	0	0	0	0	3	5
Replicate 3	0	0	0	0	2	5
Replicate 4	0	0	0	0	2	5
Σ	0	0	0	1	9	20
%	0	0	0	5	45	100

¹⁾ Corrected for substance activity

 

 

 

Table 3: 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.	≥ 8.62 mg/L	Yes

Validity criteria fulfilled:

yes

Remarks:

Please refer to table 3 at "Any other infrmation on results incl. tables"

Conclusions:

The 48 h EC50 for Daphnia magna was found to be 7.94 mg/L (based on the time weighted average using measures values) under a semi-static test design (24 h interval for water exchange).

Description of key information

EC50 (48 h): 7.94 mg a.i./L, meas. TWA (Daphnia magna, OECD 202)

Additional information

The short-term toxicity of Alcohols, C12-13, branched and linear, ethoxylated, sulfates, sodium salts to aquatic invertebrates was investigated in a OECD 202 guideline study. Daphnia magna were exposed to nominal test substance concentrations of 6.0, 8.00, 4.00, 2.00, and 1.00 mg a.i./L in a semi-static test. Test solutions were renewed after 24 h. The test concentrations were analytically monitored by LC-MS/MS. Six constituents of the UVCB substance were targeted in the analysis. The time weighted test concentrations were 15.3, 8.07, 4.02, 1.96 and 0.960 mg/L after 48 h. The determined EC50 (48 h) was 7.94 mg/L (meas. TWA).