Bis[N-(2-hydroxyethyl)-N-methylglycinato-N,O,ON]copper

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:

2017-08-08 to 2017-10-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Bis[N-(2-hydroxyethyl)-N-methylglycinato-N,O,ON]copper

EC Number:

300-491-4

EC Name:

Bis[N-(2-hydroxyethyl)-N-methylglycinato-N,O,ON]copper

Cas Number:

93940-93-3

Molecular formula:

C10H20CuN2O6

IUPAC Name:

bis[N-(2-hydroxyethyl)-N-methylglycinato-N,O,ON]copper

Test material form:

solid: particulate/powder

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: analytical samples taken from control and all test item concentrations at 0 hours (initial value) and 48 hours aged test solutions were analysed
- Fortified samples: a stock solution (1770 mg/L, purity not considered) and a dilution (1 mg/L) were prepared in acetonitrile/water (1:1, v/v) +0.1% acetic acid and used for fortification of recovery samples.
- Sample storage (before test): stored deep-frozen until analysis

Test solutions

Vehicle:

no

Details on test solutions:

- Stock solution of 15 mg/L. Lower test concentrations were prepared by dilution.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: straus, clone 5
- Feeding during test: no feeding during test
- Feeding of culture (before test): three times a week with single cell green algae
- Main test: freshly hatched daphnids less than 24 hours old were used

Study design

Test type:

static

Water media type:

freshwater

Remarks:

Elendt M4 medium

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

250 mg/L CaCO3

Test temperature:

20.2 – 21.7 °C

pH:

7.28 – 8.03

Dissolved oxygen:

≥ 8.3 mg/L

Nominal and measured concentrations:

Nominal concentrations: 1.50, 0.500, 0.167, 0.0556 and 0.0185 mg/L and a control
Measured concentration (geometric mean based): 1.70, 0.515, 0.170, 0.0556, 0.0189 mg/L and not determined for the control

Details on test conditions:

TEST SYSTEM
- Test vessel: glass vessel
- Type: covered with a glass plate (to reduce evaporation)
- Material, size, headspace, fill volume: 100 mL capacity filled with 50mL of test solution
- No. of organisms: 20 daphnids per treatment group (4 replicates of 5 animals each)
- No. of treatment groups: 6 (5 dose rates of the test item and a control)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4
- Culture medium different from test medium: same as Elendt M4

OTHER TEST CONDITIONS
- Exposure to light: none

EFFECT PARAMETERS MEASURED: Immobilisation

RANGE-FINDING STUDY
- Test concentrations: control, 100, 10 and 1 mg/L
- Sampling points : after 24h and 48h of exposure
- Results used to determine the conditions for the definitive study:
-- After 24 h 95 % immobilisation was observed at 10.0 mg/L. 100 % of the daphnids were found to be immobile at the highest test item concentration of 100 mg/L.
-- After 48 hours 100 % immobilisation was observed at 10.0 mg/L and 100 mg/L test item.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate, tested at 1.00 and 2.00 mg/L with 20 test organisms per test concentration, at around the same time period as the study

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.114 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.056 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

see any other information on results

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Test concencentrations: 1 and 2 mg/L (20 organisms per test concentration)
- Relevant effect levels: EC50 (24h) is between 1 and 2 mg/L. The results are in accordance with the requirements of the OECD guideline 202.

Any other information on results incl. tables

Results non-GLP range finding test

After 48h

	nominal test concentration (mg/L)
	control	1	10	100
group 1	0	0	5	5
group 2	0	0	5	5
group 3	0	1	5	5
group 4	0	0	5	5
sum	0	1	20	20
%	0	5	100	100

Summary of biological results:

a) Results of the test, 24h values

	nominal test item concentration [mg/L]
	Control	0.0185	0.0556	0.167	0.500	1.50
	immobilised daphnids after 24 h
Group 1	0	0	0	2	2	5
Group 2	0	0	1	1	3	5
Group 3	0	0	0	2	2	5
Group 4	0	0	1	2	5	5
Σ	0	0	2	7	12	20
%	0	0	10	35	60	100

b) Results of the test, 48h values

	nominal test item concentration [mg/L]
	Control	0.0185	0.0556	0.167	0.500	1.50
	immobilised daphnids after 48 h
Group 1	0	0	0	4	5	5
Group 2	0	0	1	5	5	5
Group 3	0	0	0	4	5	5
Group 4	0	0	1	4	5	5
Σ	0	0	2	17	20	20
%	0	0	10	85	100	100

Determined concentration of test substance:

Test item	Sampling	Test item found		Test item concentration measured
nominal [mg/L]	[h]	[mg/L]	% of nominal	[mg/L]
Control	0 fresh	< LOD	-	-
	48aged	< LOD	-	-
0.0185	0 fresh	0.0187	101	0.0189
	48 aged	0.0192	104
0.0556	0 fresh	0.0558	100	0.0556
	48 aged	0.0564	101
0.167	0 fresh	0.175	105	0.170
	48 aged	0.166	99
0.500	0 fresh	0.528	106	0.515
	48 aged	0.498	100
1.50	0 fresh	1.60	107	1.70
	48 aged	1.79	119

Effect concentrations:

	nominal concentrations		measured concentrations
	24h	48h	24h	48h
NOEC	0.0556	0.0556	0.0556	0.0556
EC50	0.269	0.102	0.335	0.114
95% confidence limit of EC50	0.186-0395	0.079-0.132	0.225-0.475	0.090-0.139

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48-hour EC50 for Daphnia magna is 0.114 mg/L (measured, geom. mean).

Executive summary:

The short-term toxicity to Daphnia magna was determined according to OECD 202 guideline with no deviations. The study was conducted under static conditions. The daphnids were exposed over a period of 48 hours to a nominal concentrations of 1.50, 0.500, 0.167, 0.0556 and 0.0185 mg/L and a control. This corresponds to 1.70, 0.515, 0.170, 0.0556, 0.0189 mg/L (geometric mean of measured concentrations).

The validity criteria were fulfilled.

The EC50 value was determined to be 0.114 mg/L (measured, geom. mean) and the NOEC 0.0556 mg/L.