Onium compounds, morpholinium, 4-ethyl-4-soya alkyl, Et sulfates

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 - 23 Nov 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

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)

Remarks:

The department of health of the government of the united kingdom, Brixham, UK

Test material

Test material information

Constituent 1

Reference substance name:

Onium compounds, morpholinium, 4-ethyl-4-soya alkyl, Et sulfates

EC Number:

263-167-0

EC Name:

Onium compounds, morpholinium, 4-ethyl-4-soya alkyl, Et sulfates

Cas Number:

61791-34-2

Molecular formula:

Not available for UVCB substance

IUPAC Name:

4-ethyl-4-(hexadecanoyloxy)morpholin-4-ium 4-ethyl-4-[(9E)-octadec-9-enoyloxy]morpholin-4-ium 4-ethyl-4-[(9E,12E)-octadeca-9,12-dienoyloxy]morpholin-4-ium triethyl sulfate

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: At the 0 hour sampling occasion, an aliquot was removed from each of the excess dosing solutions (three aliquots for 0.031 mg/L) and an equivalent volume of tetrahydrofuran (THF) was added (resulting in a x2 dilution). The sample was mixed and an aliquot transferred to an LCMS vial for analysis. At the 48 hour sampling occasion, an aliquot was removed from one test vessel from each concentration (three test vessels for 0.031 mg/L) and an equivalent volume of THF was added (resulting in a x2 dilution). The sample was mixed and an aliquot transferred to an LCMS vial for analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A primary stock concentrate of test item, with a nominal concentration of 10 mg/L, was prepared by adding a nominal 0.01 g (actual weight: 0.01002 g) of test substance to 1000 mL of culture medium. The stock was then stirred for 20 minutes. The test concentrations were prepared by diluting the stock solution and stirring of 5 min.
- Eluate: no
- Differential loading: no
- Controls: yes, test medium only
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): The resultant stock was observed to be clear and colourless with bubbles on the surface and was used to prepare the test solutions. The final test item solutions were observed as clear and colourless.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Age of test species at test start: <24 hours old
- Source: Laboratory cultures held at the laborator.
- Age of parental stock (mean and range, SD): 11 +/- 1 d
- Feeding during test: no

CULTURING CONDITIONS
- Temperature: 20 ± 2°C
- Vessel: in 2 L glass vessels with a working volume of 1.6 L
- Photoperiod: 16 h light: 8 h dark with 20-min transition period
- Culturing conditions: same as test
- Type and amount of food: Mixed algae diet of Chlorella vulgaris, strain CCAP 211/12 and Pseudokirchneriella subcapitata, strain CCAP 278/4.
- Feeding frequency: daily ad libitum depending on age and density of the culture
- Health during cultruing: The test organisms and the cultures from which they were obtained showed no evidence of disease before the test period.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Test temperature:

19.6 - 20.5

pH:

7.90 - 8.36

Dissolved oxygen:

9.04 - 9.33 mg/L

Nominal and measured concentrations:

Nominal concentrations: control, 0.010, 0.031, 0.098, 0.313 and 1.0 mg/L.
Mean measured concentrations: control, 0.0055, 0.011, 0.042, 0.15 and 0.71 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: 250 mL glass beakers
- Type: covered with glass lids
- Material, size, headspace, fill volume: glass beakers: 250 mL, headspace: 50 mL, fill volume: 200 mL
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Intervals of water quality measurement: pH and dissolved oxygen concentrations were measured at the start (measured from the excess solution remaining after filling the test vessel) and at the end of the test (measured in one replicate of each concentration and the control). Temperature was measured continually.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: according to guideline
- Culture medium different from test medium: no (Elendt's M4 D. magna medium)

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light:8 hours dark, with 20 minute dusk and dawn transition periods - Culture medium different from test medium: no (Elendt's M4 D. magna medium)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobility at 24 and 48 h. Any overt symptoms of toxicity were also recorded.

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Test concentrations: nominal test concentrations of 0.01, 0.1, 1.0, 10 and 100 mg/L (non-GLP range-finding test).
- Results used to determine the conditions for the definitive study: 48 hour observations: 0.01 mg/L: 1/10 Daphnia on surface of solution; 0.1 mg/L: 3/10 Daphnia on surface of solution. Therefore a 1 mg/L test concentration was selected as the highest concentration for the definitive test.

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.09 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other:

Remarks:

CL 95%: 0.063 - 0.10 mg/L

Details on results:

- Mortality of control: no
- Other biological observations: no other symptoms of toxicity observed

Any other information on results incl. tables

The measured concentrations at the start of the study were 100-110% and at the end 0-46% of nominal. Therefore, the mean measured concentrations were used for the calculation and reporting of the effect values.

Table 1: Daphnia magna response

Time (h)	Nominal concentration (mg/L)	Mean measured concentration (mg/L)	Number immobilised per replicate				Total number tested	Total number immobilised	Percentage immobilised
			A	B	C	D
24	Control	0	0	0	0	0	20	0	0
	0.010	0.0055	0	0	0	0	20	0	0
	0.031	0.011	0	0	0	0	20	0	0
	0.098	0.042	0	0	0	0	20	0	0
	0.313	0.15	5	5	5	4	20	19	95
	1.0	0.71	5	5	5	5	20	20	100
48	Control	0	0	0	0	0	20	0	0
	0.010	0.0055	0	0	0	0	20	0	0
	0.031	0.011	0	0	0	0	20	0	0
	0.098	0.042	0	0	2	0	20	2	10
	0.313	0.15	5	5	5	5	20	20	100
	1.0	0.71	5	5	5	5	20	20	100

Table 2: Validity criteria

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.	dissolved oxygen concentration remained at >= 3 mg/L	yes

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

please refer to "any other information on results incl. tables"