L-Glutamic acid, N-coco acyl derivs., compds. with triethanolamine (1:1)

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 February 2012 to 02 March 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Reason / purpose for cross-reference:

other: read-across target

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF 2-7-1 The guidelines related to the study reports for the registration application of pesticide (Ref. No. 12-Nousan-8147 on 24 November 2000)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All test concentrations and the untreated control were sampled for analysis.
- Sampling method: Concentrations were analytically determined at the start and at the end of the first and the last renewal period.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Prior to treatment, a stock solution of 200 mg/L was prepared by diluting an appropriate amount of test material in aquarium water. The test solutions were prepared by appropriate dilution of this stock solution, see Table 'Preparation of Test Solutions'. Test solutions were freshly prepared prior to each renewal period.
- Controls: The dilution water (circulated and filtered) was used without addition of the test material.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Common Carp
- Length at study initiation: 3.6 – 4.8 cm
- Feeding during test: The fish were not fed during the test.

ACCLIMATION
- Acclimation period: > 12 days.
- Health during acclimation (any mortality observed): Fish were observed during the acclimatisation and test periods, the fish were healthy. There was no mortality of the population for seven days before the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

The hardness of the dilution water was determined as 211 mg/L (as CaCO3).

Test temperature:

20.8 - 21.4 °C

pH:

8.16 – 8.26

Dissolved oxygen:

87 – 96 % of the air saturation value at the temperature used

Nominal and measured concentrations:

Nominal concentrations of 19.1, 34.3, 61.7, 111.1 and 200 mg/L.
Measured geometric mean concentrations of 17.7, 34.4, 56.7, 128.5 and 239.4 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquarium.
- Fill volume: 20 L of test solution.
- Renewal rate of test solution: 24 hours.
- No. of organisms per vessel: 7 per test group.
- No. of vessels per concentration (replicates): One aquarium per concentration.
- No. of vessels per control (replicates): One aquarium.
- Biomass loading rate: < 1.0 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Aquarium water, not specified.
- Intervals of water quality measurement: The water temperature, oxygen concentration and pH were measured at the start and at the end of each renewal period in each test aquarium.

OTHER TEST CONDITIONS
- Photoperiod: The light-dark cycle during the test was 16 hours light and 8 hours darkness.

EFFECT PARAMETERS MEASURED:
The observations of fish were carried out in the following intervals: at 3, 6, 24, 48, 72 and 96 hours. Mortality and any sub-lethal effect were determined in each observation period.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study : A preliminary concentration range-finding test was conducted to determine the approximate toxicity of the test material so that appropriate test concentrations can be selected for use in the definitive test. Four fish were exposed to each concentration of the test item plus a control, for 96 hours.
During the formulation procedure the test solutions were prepared by similar method as described for the definitive test.
- Test concentrations: 0.0 (untreated control, 0.1, 1, 10 and 100 mg/L.
- Results used to determine the conditions for the definitive study: One mortality was observed at the high dose level, 100 mg/L. Since a significant toxic response was observed during the preliminary range-finding test, five test concentrations in a geometric series with a separation factor of 1.8 and one control were used in the main semi-static test.
The following nominal concentrations were tested: 19.1, 34.3, 61.7, 111.1 and 200 mg/L.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

84.4 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95 % Confidence Limits 52.27 - 162.50 mg/L.

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

56.7 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

128.5 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: No sub-lethal effects were observed in the control and treated groups for a period of 96 hours.
- Mortality of control: No mortality was observed in the control vessels.

Reported statistics and error estimates:

The 24, 48, 72 and 96 hours LC50 values were calculated by Probit analysis with 95 % confidence limits using using a statistical software package (based on the calculated geometric mean concentrations).

Sublethal observations / clinical signs:

FORMUALTION ANALYSIS

- The measured geometric mean test item concentrations were: 17.7, 34.4, 56.7, 128.5 and 239.4 mg/L.

- As the measured concentrations deviated more that 20 per cent from the nominal in one cases, biological results are based on measured geometric mean concentrations.

 

Analytical Results

Renewal	Test Concentration (mg/L)	Measured Concentration at the Start (95 % confidence Intervals)		Measured Concentration at the Start (95 % confidence Intervals)
		mg/L	% of Nominal	mg/L	% of Nominal
First Renewal	Control	Not Detected	-	-	-
	19	18.7 ± 4.0	99	23.4 ± 7.8	123
	34.3	38.9 ± 2.9	113	38.0 ± 2.6	111
	61.7	60.7 ± 2.5	98	56.6 ± 2.2	92
	111.1	127 ± 7.5	114	130 ± 7.6	117
	200	232 ± 23.9	116	247 ± 16.2	124
Second Renewal	Control	Not Detected	-	-	-
	19	18.3 ± 3.0	96	12.2 ± 2.1	64
	34.3	34.9 ± 3.5	102	27.3 ± 2.8	80
	61.7	59.1 ± 0.8	96	51.0 ± 1.9	83

 

Cumulative mortality data in the definitive test

Concentrations (mg/L)		Cumulative Mortality (No. of fish) Initial Population of 7 Fish
		3 Hour	6 Hours	24 Hours	48 Hours	72 Hours	96 Hours
Nominal	Measured	0	0	0	0	0	0
Control	0.0	0	0	0	0	0	0
19.1	17.7	0	0	0	0	0	0
34.3	34.4	0	0	0	0	0	0
61.7	56.7	0	0	0	0	0	0
111.1	128.5	0	0	4	7	7	7
200	239.4	7	7	7	7	7	7

 

Validity criteria

The study fulfilled all validity criteria outlined in the most recent EU guidance (OECD 203, 2019) as detailed below:

• Control mortality should be less than or equal to 10% (or 1 fish if less than 10 control fish are tested) by the end of the test. 0% control mortality was observed in the study.


• Dissolved oxygen concentrations should be greater than or equal to 60% of the air saturation value in all test vessels throughout the exposure. greater than or equal to 87% was observed throughout the exposure period.

• Analytical measurement of test concentrations was completed. As the concentration values measured in all test concentrations at the start and at the end of the first and at the last renewal periods deviated more that 20% from the nominal in one cases (see Table below). Therefore, the, biological results are based on measured geometric mean concentrations.

Validity criteria fulfilled:

yes

Remarks:

See 'Any other information on results incl. tables' for further information.

Conclusions:

Under the conditions of the test, mortality was observed at measured concentrations of ≥ 128.5 mg/L. No sub-lethal effects were observed during exposure.
Based on these results it was possible to calculate the following effect parameters; 96 hour LC50 84.4 mg/L (95 % confidence limits 52.27 - 162.50 mg/L); NOEC 56.7 mg/L; LOEC 128.5 mg/L.

Executive summary:

In a read-across study the acute toxicity of the test material to fish was determined in an acute toxicity study using Cyprinus carpio. The study was performed under GLP conditions and in line with the standardised guidelines OECD 203, EU Method C.1, EPA OPPTS 850.1075, and Japanese MAFF (2-7-1).

Based on the results of the preliminary range-finding test 7 fish per concentration were exposed for 96 hours under semi-static conditions to nominal test material concentrations of 19.1, 34.3, 61.7, 111.1 and 200 mg/L. Corresponding to measure geometric mean concentration of 7.7, 34.4, 56.7, 128.5 and 239.4 mg/L. Since measured values deviated more than 20 % of the nominal, effect parameters were expressed in terms of measured concentrations.

Under the conditions of the test, mortality was observed at measured concentrations of ≥ 128.5 mg/L. No sub-lethal effects were observed during exposure. Based on these results it was possible to calculate the following effect parameters; 96 hour LC50 84.4 mg/L (95 % confidence limits 52.27 - 162.50 mg/L); NOEC 56.7 mg/L; LOEC 128.5 mg/L.

Description of key information

Under the conditions of the test, mortality was observed at measured concentrations of ≥ 128.5 mg/L. No sub-lethal effects were observed during exposure. Based on these results it was possible to calculate the following effect parameters; 96 hour LC50 84.4 mg/L (95 % confidence limits 52.27 - 162.50 mg/L); NOEC 56.7 mg/L; LOEC 128.5 mg/L.

.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

84.4 mg/L

Additional information

Read-Across Substance: Sipos (2013c)

The acute toxicity of the test material to fish was determined in an acute toxicity study using Cyprinus carpio. The study was performed under GLP conditions and in line with the standardised guidelines OECD 203, EU Method C.1, EPA OPPTS 850.1075, and Japanese MAFF (2-7-1).

Based on the results of the preliminary range-finding test 7 fish per concentration were exposed for 96 hours under semi-static conditions to nominal test material concentrations of 19.1, 34.3, 61.7, 111.1 and 200 mg/L. Corresponding to measure geometric mean concentration of 7.7, 34.4, 56.7, 128.5 and 239.4 mg/L. Since measured values deviated more than 20 % of the nominal, effect parameters were expressed in terms of measured concentrations.

Under the conditions of the test, mortality was observed at measured concentrations of ≥ 128.5 mg/L. No sub-lethal effects were observed during exposure. Based on these results it was possible to calculate the following effect parameters; 96 hour LC50 84.4 mg/L (95 % confidence limits 52.27 - 162.50 mg/L); NOEC 56.7 mg/L; LOEC 128.5 mg/L.