Imidazolium compounds, 2-(C9-19 and C9-19-unsatd. alkyl)-1-[(C10-20 and C10-20-unsatd. amido)ethyl]-4,5-dihydro-1-Me, Me sulfates

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:

From September 12, 2017 to September 15, 2017

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:

not specified

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

High-performance liquid chromatography (HPLC)

Details on sampling:

To demonstrate that nominal exposure concentrations were being achieved the concentrations of test substance in the test vessels were measured using the high-performance liquid chromatography method. At the start of the test additional sacrificial vessels with dispensed test solution were taken for whole sample analysis and at the end of the test, replicate vessels of the dilution water control, solvent control and each test concentration was taken for whole sample analysis.

Vehicle:

yes

Remarks:

Tetrahydrofuran

Details on test solutions:

The study was run with a dilution water control, solvent control and nominal exposure concentrations of 0.018, 0.032, 0.056, 0.1, 0.18 and 0.32 mg/L. A primary stock concentrate of test substance, with a nominal concentration of 3.2 g/L, was prepared by adding a nominal 0.032 g of test substance to 10 mL of the solvent tetrahydrofuran (THF). The resultant stock was observed to be clear and colourless and was used to prepare the test solutions. This was achieved by the direct addition of the appropriate amount of concentrate to dilution water via a microliter syringe into a stirring solution in a volumetric flask. The solvent control was prepared in the same way using solvent only. The control consisted of culture medium only. A solvent was used in this study to assist in dosing the test compound due to its apparent low solubility in test media, the concentration of solvent used in all exposure solutions with the exception of the control was 100 µg/L. All test solutions were clear and colourless.

Test organisms (species):

Daphnia magna

Details on test organisms:

The test organism was the freshwater crustacean, Daphnia magna, obtained from continuous laboratory cultures held at Scymaris. The stock cultures of D. magna were maintained in a reconstituted water medium, the same as the test dilution water, at a temperature of 20 ± 2°C. The cultures were maintained in 2 L glass vessels with a working volume of 1.6 L. A photoperiod of 16 h light:8 h dark, with 20-minute transition periods was provided. The D. magna cultures were fed on a mixed algae diet of Chlorella vulgaris, strain CCAP 211/12 and Pseudokirchneriella subcapitata, strain CCAP 278/4. The D. magna cultures were fed daily ad libitum depending on age and density of the culture. Culture conditions were such that the D. magna reproduction was by diploid parthenogenesis. D. magna <24 h old, obtained from a single culture vessel, were used for testing. The parent animals were 25 ± 1 d old and had been maintained with a twice weekly renewal of reconstituted water medium since birth. The test organisms and the culture from which they were obtained showed no evidence of disease before the test period.

Test type:

static

Water media type:

other: Elendt's M4 D. magna medium

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20±1°C

pH:

7.80 to 7.95

Dissolved oxygen:

8.63 to 8.95 mg/L

Nominal and measured concentrations:

Control, solvent control and 0.018, 0.032, 0.056, 0.1, 0.18 and 0.32 mg/L (nominal)
Control, solvent control and 0.014, 0.024, 0.043, 0.042, 0.097 and 0.17 mg/L (measured)

Details on test conditions:

Apparatus
Glass beakers of 250 mL nominal capacity were used as test vessels, with four replicates per test substance concentration. Each vessel contained 200 mL of test solution providing a depth of approximately 60 mm. The beakers were covered with loose fitting glass lids. The positions of the treatments were randomly allocated within the test area.

Reference substance (positive control):

no

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 0.17 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 0.1 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Remarks on result:

other: Confidence interval: 0.086 – 0.122 mg/L

Remarks:

Linear regression (GLM) method

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

ca. 0.097 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

ca. 0.043 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Details on results:

Analytical data
The limit of quantification of test substance in this study was 0.0020 mg/L. All analytical values are quoted to two significant figures and percentages to the nearest integer. The measured concentrations at the start of the study were 54-99% of nominal and at the end were 29-63% of nominal. On the basis of the analytical data the mean measured concentrations were used for the calculation and reporting of results.

Biological data
Based on immobility compared to the control (p <0.05) the 48 h No Observed Effect Concentration (NOEC) was determined to be 0.043 mg/L and the Lowest Observed Effect Concentration (LOEC) was 0.097 mg/L. There was no immobility observed in the dilution water control. No other symptoms of toxicity were observed.

Analytical results

Nominal concentration of Quaternium87 (mg/L)	Measured concentration of Quaternium 87  (mg/L)				Mean measured concentration (mg/L)	Mean measured concentration (%)
	0 h		48 h
	(mg/L)	% of nominal	(mg/L)	% of nominal
Control	0	-	0	-	0	-
Solvent Control	0	-	0	-	0	-
0.018	0.017	95	0.011	61	0.014	78
0.032	0.028	89	0.020	63	0.024	76
0.056	0.055a	99	0.032b	57	0.043	78
0.1	0.054	54	0.029	29	0.042	42
0.18	0.13	74	0.061	34	0.097	54
0.32	0.23	73	0.11	33	0.17	53

All measurements are quoted to 2 significant figures

a Mean of triplicate analyses:0.053, 0.057, 0.055mg/L

b Mean of triplicate analyses:0.030, 0.032, 0.033mg/L

The limit of quantification in the study was 0.0020mg/L

Daphnia magna response

Time (h)	Nominal concentration of Quaternium 87 (mg/L)	Number immobilised per replicate				Total number tested	Total number immobilised	Percentage immobilised
		A	B	C	D
24	Control	0	0	0	0	20	0	0
	Solvent Control	0	0	0	0	20	0	0
	0.0.18	0	0	0	0	20	0	0
	0.032	0	0	0	0	20	0	0
	0.056	0	0	0	0	20	0	0
	0.1	0	0	0	0	20	0	0
	0.18	0	0	0	0	20	0	0
	0.32	0	0	0	0	20	0	0
48	Control	0	0	0	0	20	0	0
	Solvent Control	0	0	0	0	20	0	0
	0.0.18	0	0	0	0	20	0	0
	0.032	0	0	0	0	20	0	0
	0.056	0	0	0	0	20	0	0
	0.1	0	0	0	0	20	0	0
	0.18	3	4	1	3	20	11	55
	0.32	4	4	5	4	20	17	85

Validity criteria

As no immobility or other signs of stress were observed in the control and dissolved oxygen concentration remained at ≥3 mg/L, this test has satisfied all OECD Guideline 202 validity criteria.

Validity criteria fulfilled:

yes

Conclusions:

Under study conditions, the 48 h EC50 was determined to be 0.10 mg/L (measured).

Executive summary:

A study was conducted to determine the acute toxicity of test substance, 'di-C16 and C18-unsatd. AAEMIM-MS' (active: 100%), to Daphnia magna, according to OECD Guideline 202, in compliance with GLP. Twenty test organisms were exposed to each nominal test substance concentrations of 0, 0.018, 0.032, 0.056, 0.1, 0.18 and 0.32 mg/L for 48 h under static conditions. The concentrations of test substance in the test vessels were measured using the HPLC method. The mean measured concentrations of test substance were determined to be 0, 0.014, 0.024, 0.043, 0.042, 0.097 and 0.17 mg/L. Immobilisation was recorded at 24 and 48 h and compared with control values. Based on immobility compared to the control (p <0.05), the 48 h NOEC and LOEC were determined to be 0.043 and 0.097 mg/L (measured) respectively. No other symptoms of toxicity were observed. Based on the study results, the 48 h EC50 was calculated to be 0.10 mg/L (measured). As no immobility or other signs of stress were observed in the control and dissolved oxygen concentration remained at ≥3 mg/L, the test was considered to have satisfied all the validity criteria. Under study conditions, the 48 h EC50 was determined to be 0.10 mg/L (measured) (Scymaris, 2017).

Description of key information

Based on the results of the read across study, the 48 h EC50 value of the test substance for toxicity to aquatic invertebrates is considered to be 0.10 mg/L (measured).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.1 mg/L

Additional information

A study was conducted to determine the acute toxicity of test substance, 'di-C16 and C18-unsatd. AAEMIM-MS' (active: 100%), to Daphnia magna, according to OECD Guideline 202, in compliance with GLP. Twenty test organisms were exposed to each nominal test substance concentrations of 0, 0.018, 0.032, 0.056, 0.1, 0.18 and 0.32 mg/L for 48 h under static conditions. The concentrations of test substance in the test vessels were measured using the HPLC method. The mean measured concentrations of test substance were determined to be 0, 0.014, 0.024, 0.043, 0.042, 0.097 and 0.17 mg/L. Immobilisation was recorded at 24 and 48 h and compared with control values. Based on immobility compared to the control (p <0.05), the 48 h NOEC and LOEC were determined to be 0.043 and 0.097 mg/L (measured) respectively. No other symptoms of toxicity were observed. Based on the study results, the 48 h EC50 was calculated to be 0.10 mg/L (measured). As no immobility or other signs of stress were observed in the control and dissolved oxygen concentration remained at ≥3 mg/L, the test was considered to have satisfied all the validity criteria. Under study conditions, the 48 h EC50 was determined to be 0.10 mg/L (measured) (Scymaris, 2017).