1-Tetradecanaminium, N,N,N-trimethyl-, ethanedioate (2:1)

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study under GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Tetradecyltrimethylammonium oxalate

IUPAC Name:

Tetradecyltrimethylammonium oxalate

Constituent 2

Reference substance name:

-

EC Number:

453-080-8

EC Name:

-

Cas Number:

154858-16-9

Molecular formula:

C36H76N2O4

IUPAC Name:

bis(trimethyl(tetradecyl)azanium) oxalate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 5.12 - 12.8 - 32.0 - 80.0 - 200 - 500 µg/L
- Sampling method: All test concentrations above LOQ and the control were analytically verified by photometer in the fresh media (at 0 hours) and old media (after 48 hours) as specified below.
Sampling from freshly prepared media was carried out immediately after homogenisation. Samples of the old media were taken directly from the test vessels.
- Sample storage conditions before analysis: All samples were stored at 7±2°C before preparation and before analysis, if necessary.

Test solutions

Vehicle:

no

Details on test solutions:

Stock solution: 50 mg/L, prepared with dilution water
Dispersion treatment: Agitation
Test concentrations: 500-200-80.0-32.0- 12.8-5.12 µg/L

In the stock solution of 50 mg/L a slight turbidity was observed at test start.
In all tested concentration levels the test item was clearly dissolved throughout exposure.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu)
- Age at study initiation: 2 to 24 h
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 21 °C, in an incubator, 16 h illumination, illumination strength max. 20 µEmol x m-2 x s-1
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding

DILUTION WATER:
Dilution water according to EC 92/69 L383A C.2

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

no

Test conditions

Hardness:

Dilution water at test start: 244 mg CaCO3/l

Test temperature:

21±1°C

pH:

7.51-7.80

Dissolved oxygen:

8.08-8.52 mg/l

Salinity:

not measured, freshwater

Nominal and measured concentrations:

please refer to "Any other information on material and methods"

Details on test conditions:

Test vessels: Glass beakers (5 cm ID x 8 cm H), 50 ml
Volume of the study medium: 20 ml
Number of study organisms per concentration and control: 20 animals, divided into 4 (2*) parallel samples, each with 5 (10*) animals
Number of animals per test vessel: 5 (10*)
Number of parallel samples per concentration: 4 (2*)
Photoperiod: 16/8 h light/dark cycle

*) The information in brackets refer to the test of the reference item

TEST DESCRIPTION
A static preliminary range finding tests was conducted with 6 concentrations of the test item ranging from 0.5 to 50 000 µg/L, set up in a geometric series with a factor of 10 (NON-GLP-state ).

Based on the results of the preliminary test the definitive test was performed with 6 concentrations ranging from 5.12 to 500 µg/L in a geometric series with a factor of 2.5 under static conditions to enable the determination of immobilisation after 24 and 48 hours.

2 to 24 h old daphnids were used for the definitive test. They were obtained by removing the mother animals twice within 22 h. The daphnids were acclimated to the dilution water ca. 2 h before the test started. The daphnids were not fed during the study.

Control replicates with dilution water (test medium without test item) were tested under the same conditions as the test groups.

20 daphnids divided into 4 (2*) groups with 5 (10*) animals per test vessel were used. All test beakers were placed in an incubator (light/dark cycle 16/8 for 48 h at 20 ± 1 °C constant). In the definitive test the immobilisation of the daphnids was observed after 24 and 48 hours of exposure. Water parameters were measured at begin and end of the study.

The reference item was tested in five concentrations within the concentration range 0.58 to 5.8 mg/L.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

29.3 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

37.9 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

12.8 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

32 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: 0 %
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No

Results with reference substance (positive control):

The percentage immobility for the reference item was determined after 24 h. The EC100-value was determined directly from the test results. The EC10 and EC50- with 95 % confidence interval (CI) was determined in a probability network by interpolation according to standard procedures.

EC-values after 24 h of the reference item in mg/L:
EC10 : 0.96
EC50 : 1.73 (CI 1.47- 2.03)
EC100 : 5.80
The EC50-value of the reference item potassium dichromate after 24 h is within the prescribed concentration range of 1.0 - 2.5 mg/L of quality criteria according to AQS: DIN Guideline 38412 L 30.

Reported statistics and error estimates:

The No Observed Effect Concentration (NOEC) for immobilisation (<10 %) of 12.8 µg/L was determined directly from the test results obtained.

The concentration-effect-relationships after 24 and 48 h of exposure are graphically illustrated in a probability network. The 24 and 48 h EC10and EC50-values were calculated by probit analysis.

Any other information on results incl. tables

Percentage of Daphnids Incapable of Swimming after 24 and 48 h of Exposure (n = 20)

Nominal	IMMOBILIZATION [%]
Test Item Concentration	24 h					48 h
[µg/L]	Replicates					Replicates
	1	2	3	4	MV	1	2	3	4	MV
500	100	100	100	100	100	100	100	100	100	100
200	80	100	100	100	95	100	100	100	100	100
80.0	60	40	20	60	45	100	100	100	100	100
32.0	0	0	20	0	5	0	20	40	20	20
12.8	0	0	20	0	5	0	20	20	0	10
5.12	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item the 48-h EC50 for Daphnia magna was calculated to be 37.9 µg/L (95 % confidence limits: 34.7 – 41.4 µg/L). The NOEC after 48 h was 12.8 µg/L. The LOEC after 48 h was 32.0 µg/L.

Executive summary:

The Acute lmmobilisation (EC50) of the test item, to Daphnia magna (STRAUS) was determined according to OECD Guideline 202 (1999, Draft) and EC Directive 92/69/EC Method C.2 (1992) from 2003-03-10 to 2003-03-12 at DR.U.NOACK-LABORATORIEN, D-31157 Sarstedt, Germany. The study was conducted under static conditions with 6 concentrations ranging from 5.12 to 500 µg/L in a geometric series with a factor of 2.5 over a duration of 48 hours. 20 test organisms were exposed to each test concentration and control. The test concentration levels of 12.8 to 500 µg/L of the test item and the control were analytically verified via photometer after 0 h (new media) and 48 h (old media). Recovery rates of the test item were > 80 %. All results are given based on the nominal test item concentrations. Details of the analytical results are given in part 9. The EC10 and

EC50-values were calculated by probit analysis according to WEBER (1986). A reference test was carried out with potassium dichromate as reference item. The EC50-value of the reference item at 1. 73 mg/L after 24 h was within the prescribed concentration range of 1.0 to 2.5 mg/L according to AQS, DIN Guideline 38412 L 30. Water quality parameters of pH-value and dissolved oxygen concentration measured at 0 h and 48 h were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled.

EC10, EC50- (with Confidence Interval), EC100 -Values, NOEC and LOEC (based on nominal concentrations)

	Test	Nominal Test Item Concentration
	Duration	[µg/L]
	[h]
EC10	24	40.1
	48	29.3
EC50	24	81.9
	48	37.9
95 % confidence interval	24	67.1-99-9
	48	34.7-41.4
EC100	24	500
	48	80.0
NOEC	48	12.8
LOEC	48	32.0