1-Propanaminium, 2-hydroxy-N-(2-hydroxyethyl)-N-methyl-N-[2-[(1-oxo-9-octadecen-1-yl)amino]ethyl]-3-sulfo-, methyl sulfate, sodium salt (1:1:1)

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:

22.10.2018 - 30.11.2018

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:

no

Vehicle:

no

Details on test solutions:

A water accommodated fraction (WAF) equilibration trail was performed prior to exposure. The purpose of the equilibration trail is to determine the optimum mixing duration to be used for WAF preparation. One WAF was prepared at each of three loading levels 1.0, 50.0 and 100 mg/L. At 2, 4, 6 and 24 hours after initiation of mixing, mixing was stopped and the solutions were allowed to settle for one hour. A sample of WAF was removed from each loading level and mixing was resumed. The concentration of organic carbon that had solubilized into the WAF from the test substance was measured using the TOC analyzer (TOC Analyzer/TNML; Make: Shimadzu; Model: LCPH E200 ROHS).

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Strain: Straus
- Source: Environmental and Molecular Toxicology Division, Department of Zoology, Karnataka University, Karnataka, India
- Feeding during test: no

ACCLIMATION
- Acclimation period: 48h
- Acclimation conditions: same as test conditions
- Type and amount of food: live algal cells (Pseudokirchneriella subcapitata) of 2 mL per liter
- Health during acclimation: no mortality

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

216 mg CaCO3/L

Test temperature:

20.1 to 21.3°C

pH:

7.16 to 7.83

Dissolved oxygen:

7.89 to 8.44 mg/L

Nominal and measured concentrations:

Nominal concentrations: 3.4, 7.5, 16.5, 36.4 and 80 mg/L
no measured concentrations

Details on test conditions:

TEST SYSTEM
- Test vessel: glass jars
- Material, fill volume: glass, 100 mL
- Aeration: no
- Renewal of test media: after 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 1

TEST MEDIUM PARAMETERS
- Preparation of dilution water: reconstituted water according to OECD 202 (with Calcium chloride, Potassium chloride, Magnesium sulfate and Sodium bicarbonate)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours of light and 8 hours dark cycle

EFFECT PARAMETERS MEASURED:
Each test vessel was observed for immobilized Daphnids at 24 and 48 hours after the beginning of the test.
Total hardness of the diluent water (reconstituted water) prior to its use for exposure was analyzed. Temperature, pH and dissolved oxygen were recorded at the beginning and at end of the test in control and in all the tested concentrations.

RANGE-FINDING STUDY
- Test concentrations: 0.01, 0.1, 1.0, 10.0, 50 and 100 mg/L
- Results used to determine the conditions for the definitive study: The immobilization percentages of 30, 80 and 100 % were recorded at the nominal loading rates of 10.0, 50.0 and 100.0 mg/L, respectively, during the 48 hour observation period.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

3.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No clinical signs of toxicity or immobilisation were observed in control and at the nominal loading rate of 3.4 mg/L during the 48 hour observation period. Daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 7.5, 16.5 and 36.4 mg/L during the 24 and 48 hour observation period. Daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 80.0 mg/L during the 24 hour observation period. The immobilization percentages of 15, 40, 70 and 100 % were recorded at the tested nominal loading rates of 7.5, 16.5, 36.4 and 80.0 mg/L, during the 48 hour observation period.

Results with reference substance (positive control):

The 48 hour EC50 of the reference substance Potassium dichromate on Daphnia magna was determined to be 0.57 mg/L. Thus, the 48 hour EC50 of the reference substance was within the acceptance range of the guideline (0.50 to 2.12 mg/L) confirming the suitability of the test system.

Table1: Summary of immobilization at different time points during main study

Group	Nominal Loading Rate (mg/L)	R	Initial No. of Daphnids	No. of Daphnids immobilized		Cummulative No.		% of Immobilisation
				24 h	48 h	24 h	48 h	24 h	48 h
G1	0	R1	5	0	0	0	0	0	0
		R2	5	0	0
		R3	5	0	0
		R4	5	0	0
G2	3.4	R1	5	0	0	0	0	0	0
		R2	5	0	0
		R3	5	0	0
		R4	5	0	0
G3	7.5	R1	5	0	1	1	3	5	15
		R2	5	0	0
		R3	5	1	2
		R4	5	0	0
G4	16.5	R1	5	1	3	4	8	20	40
		R2	5	1	3
		R3	5	1	1
		R4	5	1	1
G5	36.4	R1	5	1	3	7	14	35	*70
		R2	5	2	4
		R3	5	2	3
		R4	5	2	4
G6	80.0	R1	5	3	5	12	20	60	*100
		R2	5	2	5
		R3	5	4	5
		R4	5	3	5

 h: hour; R: Replicate; *: Statistical significance of >0.05.

Table 2: TOC analysis during main study

		TOC During Exposure (mg)
Group	Nominal Loading rate (mg/L)	Day 0 (Fresh)	Day 1 (Aged)	Day 1 (Fresh)	Day 2 (Aged)
G1	0.0	-	-	-	-
G2	3.4	2.1210	1.960	1.927	1.822
G3	7.5	2.5200	2.104	2.292	2.076
G4	16.5	4.7940	3.831	4.149	4.100
G5	36.4	8.6140	7.623	10.120	9.813
G6	80.0	20.070	19.780	24.260	24.440

Validity criteria fulfilled:

yes

Conclusions:

The 48-hour EL50 of the test item on Daphnia magna was determined to be 18 mg/L.

Executive summary:

The test item was evaluated for acute toxicity on Daphnia magna as per the OECD Guideline No. 202, “Daphnia sp., Acute Immobilisation Test”. Daphnids were exposed for the test item by aqueous exposure (treatment) under static condition. Daphnidswere observed at 24 and48 hours for signs of toxicity and immobilisation. Range Finding Study: No clinical signs of toxicity or immobilization were observed in control and at the nominal loading rates of 0.01, 0.1, 1.0 and 10.0 mg/L during the 48 hour observation period. Clinical sign of lethargy and localized on bottom of container was observed at 24 hour observation and clinical sign of localized on bottom of container was observed at 48 hour observation period at the nominal loading rates of 50.0 mg/L. Daphnids localized on bottom of container and lethargy was observed at the nominal loading rate of 100.0 mg/L during the 24 hour observation period. The immobilization percentages of 30, 80 and 100 % were recorded at the nominal loading rates of 10.0, 50.0 and 100.0 mg/L, respectively, during the 48 hour observation period.
Main Study: Based on the results of the range finding study, main study was conducted at the nominal loading of 3.4, 7.5, 16.5, 36.4 and 80.0 mg/L test item of test medium along with control. No clinical signs of toxicity or immobilisation were observed in control at the nominal loading rate of 3.4 mg/L during the 48 hour observation period. Daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 7.5, 16.5 and 36.4 mg/L during the 24 and 48 hour observation period. Daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 80.0 mg/L during the 24 hour observation period. The immobilization percentages of 15, 40, 70 and 100 % were recorded at the tested nominal loading rates of 7.5, 16.5, 36.4 and 80.0 mg/L, during the 48 hour observation period.Based on the results of the experiment and under experimental conditions employed, it is concluded that the 48 hour acute median Effective Lethal Loading Rate (EL50) of test item on Daphnia magna is 18.00 mg/L. The 95% confidence interval with lower limit of 14.69 mg/L and upper limit of 22.06 mg/L.The NOEC was 3.4 mg/L.

Description of key information

The 48-hour EL50 of the test item on Daphnia magna was determined to be 18 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

18 mg/L

Additional information

The test item was evaluated for acute toxicity on Daphnia magna as per the OECD Guideline No. 202, “Daphnia sp., Acute Immobilisation Test”. Daphnids were exposed for the test item by aqueous exposure (treatment) under static condition. Daphnids were observed at 24 and 48 hours for signs of toxicity and immobilisation. Range Finding Study: No clinical signs of toxicity or immobilization were observed in control and at the nominal loading rates of 0.01, 0.1, 1.0 and 10.0 mg/L during the 48 hour observation period. Clinical sign of lethargy and localized on bottom of container was observed at 24 hour observation and clinical sign of localized on bottom of container was observed at 48 hour observation period at the nominal loading rates of 50.0 mg/L. Daphnids localized on bottom of container and lethargy was observed at the nominal loading rate of 100.0 mg/L during the 24 hour observation period. The immobilization percentages of 30, 80 and 100 % were recorded at the nominal loading rates of 10.0, 50.0 and 100.0 mg/L, respectively, during the 48 hour observation period.

Main Study: Based on the results of the range finding study, main study was conducted at the nominal loading of 3.4, 7.5, 16.5, 36.4 and 80.0 mg/L test item of test medium along with control. No clinical signs of toxicity or immobilisation were observed in control at the nominal loading rate of 3.4 mg/L during the 48 hour observation period. Daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 7.5, 16.5 and 36.4 mg/L during the 24 and 48 hour observation period. Daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 80.0 mg/L during the 24 hour observation period. The immobilization percentages of 15, 40, 70 and 100 % were recorded at the tested nominal loading rates of 7.5, 16.5, 36.4 and 80.0 mg/L, during the 48 hour observation period. Based on the results of the experiment and under experimental conditions employed, it is concluded that the 48 hour acute median Effective Lethal Loading Rate (EL50) of test item on Daphnia magna is 18.00 mg/L. The 95% confidence interval with lower limit of 14.69 mg/L and upper limit of 22.06 mg/L. The NOEC was 3.4 mg/L.