N-methyl-N-[C18-(unsaturated)alkanoyl]glycine

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

30 May - 01 Jun 1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

adopted in 1982

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution contained 100 mg/L test item and 100 mg/L Tween 80 and was stirred for 60 min at 293 K. A serial dilution with dilution water was prepared and each test vessel received 10 mL of the respective dilution.
- Chemical name of vehicle: Tween 80

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Straus
- Source: Laboratory breed at BASF, originally obtained in 1978 from the Institut National de Recherche Chimique Appliquée, France
- Age at study initiation (mean and range, SD): 2 - 24 h

ANIMAL HUSBANDRY
- Light: 16 h light/8 h dark, 5 µE/m2 x s, 400 - 750 nm
- Type and amount of food: Green algae
- Feeding frequency: Once daily

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

2.77 mmol/L

Test temperature:

292.5 - 293.8 K

pH:

7.77 - 8.14

Dissolved oxygen:

8.36 - 8.87

Nominal and measured concentrations:

Nominal: 0, 0.098, 0.195, 0.39, 0.781, 1.56, 3.12, 6.25, 12.5, 25.0, 50.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Centrifuge glass vessels with flat bottom
- Material, fill volume: Glass, 10 mL
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Deionised water
- Ca/Mg ratio: 4:1
- Conductivity: 600 - 700 µS/cm
- Intervals of water quality measurement: Temperature, oxygen and pH were measured at test start and test end.

OTHER TEST CONDITIONS
- Light intensity: Diffuse light

EFFECT PARAMETERS MEASURED (with observation intervals if applicable)
- Immobilisation was recorded after 0, 3, 6, 24 and 48 h.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.53 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 0.47 - 0.59

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

0.39 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

0.781 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: No mortality
- Immobilisation: 5% immobilisation at 0.39 mg/L, 100% immobilisation at 0.781 to 50 mg/L

Reported statistics and error estimates:

Determination of EC50 (48h) by moving average method

Table 1: Immobilisation of Daphnia magna

Concentration (mg/L)	Mobile Daphnia after
	0 h	24 h	48 h
0	20	20	20
0.098	20	20	20
0.195	20	20	20
0.39	20	20	19
0.781	20	7	0
1.56	20	0	0
3.12	20	0	0
6.25	20	0	0
12.5	20	0	0
25.0	20	0	0
50.0	20	0	0

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 Jul - 24 Jul 2010

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:

adopted in 2004

Deviations:

yes

Remarks:

Higher temperature during the test (Guideline: 18 - 22 °C, Test: 23.5 - 23.7 °C)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

adopted in 2008

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Mainz, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All concentrations were measured at test start and test end.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution containing 10 g/L was prepared using the following procedure: 10 g test item was mixed with 1.1 g NaOH (45%). 40 mL deionised water (80 – 100 °C) was added. After homogenisation with a glass stirrer, 85 mL deionised water (room temperature) was added. Then, the vessel was stirred on a magnetic stirrer. 0.5 M NaOH was added until the resulting solution was clear. The pH of the solution was adjusted to 7.8 ± 0.2 with 1 M HCl. the solution was transferred into a volumetric flask and filled up to 1000 mL with deionised water. From this solution, the concentrations to be tested were prepared by diluting with dilution water.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Straus
- Source: Umweltbundesamt Berlin, Germany, received on 27 Sep 2007
- Age at study initiation (mean and range, SD): < 24 h
- Feeding during test: None

ANIMAL HUSBANDRY
- Method of breeding: Daphnia magna is bred in the LAUS GmbH throughout the year. The animals are kept in 2L glass beakers in M4 medium and multiply by parthenogenesis thus being genetically identical.
- Type and amount of food: Unicellular green algae
- Feeding frequency: Twice a week

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

250 mg CaCO3/L

Test temperature:

23.2 - 23.7 °C

pH:

7.3 - 7.8

Dissolved oxygen:

6.4 - 8.3 mg/L

Nominal and measured concentrations:

Nominal: 0, 0.1, 0.22, 0.46, 1.0, 2.2, 4.6, 10, 22, 46, 100 mg/L
Mean measured: < LOQ, < LOQ, < LOQ, < LOQ, 0.42, 0.60, 1.77, 5.58, 12.14, 28.94, 64.70 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Beakers
- Material, size, fill volume: Glass, 50 mL, 20 mL
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Deionised water was used for the preparation of M4 medium
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH and oxygen concentration were measured at test start and test end.

OTHER TEST CONDITIONS
- Photoperiod: Darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable)
- After 24 and 48h the immobilised daphnia were counted.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.43 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.38 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: 5% after 48 h
- Immobilisation: 10% at nominal 0.46 mg/L, 70% at nominal 1.0 mg/L, 100% at nominal 2.2 mg/L to 100 mg/L
- Analytical measurements: In treatments 0.1 – 0.46 mg/L (nominal) no test item was detectable after 48 h. The recovery after 48 h in treatments 1 – 100 mg/L (nominal) was in the range of 44 – 128% of the start concentration. Because of the medium correlation between nominal and measured concentrations, the determination of the biological results was based on the geometric mean of the measured concentrations for treatments 1 – 100 mg/L (nominal) and on the measured concentration at test start for treatment 0.46 mg/L (nominal, measured: 0.38 mg/L). Treatments 0.1 and 0.22 mg/L were not used for the evaluation of the results.

Results with reference substance (positive control):

- The reference test was carried out 08 - 09 Jun 2010
- EC50 (24h): 1.6 mg/L, 95% CL = 1.4 - 1.9 mg/L
- NOEC (24h): 1.0 mg/L

Immobilities Exp. 1

Nominal Conc. In mg/L	Immobility 24 hours					Immobility 48 hours
	abs.				in %	abs.				in %
0	0	0	0	0	0	0	0	0	0	0
1.0	3	3	4	2	60	4	4	4	2	70
2.2	5	5	5	5	100	5	5	5	5	100
4.6	5	5	5	5	100	5	5	5	5	100
10	5	5	5	5	100	5	5	5	5	100
22	5	5	5	5	100	5	5	5	5	100
46	5	5	5	5	100	5	5	5	5	100
100	5	5	5	5	100	5	5	5	5	100

 

Immobilities Exp. 2

Nominal Conc. In mg/L	Immobility 24 hours					Immobility 48 hours
	abs.				in %	abs.				in %
0	1	0	0	0	5	1	0	0	0	5
0.1	0	0	0	0	0	0	0	0	0	0
0.22	0	0	0	0	0	0	0	0	0	0
0.46	0	0	0	0	0	1	1	0	0	10

 

Measured concentrations

Nominal Conc. in mg/L	Measured Conc. in mg/L		% Recovery after 48h	% of Nominal Conc.		Geom. Mean in mg/L
	0 h	48 h		0 h	48 h
0	n.d.	n.d.	-	-	-	-
0.1	0.36	n.d.	0	363	--	--
0.22	0.37	n.d.	0	166	--	--
0.46	0.38	n.d.	0	82	--	--
1	0.51	0.35	70	51	35	0.42
2.2	0.90	0.40	44	41	18	0.60
4.6	2.20	1.43	65	48	31	1.77
10	5.80	5.36	93	58	54	5.58
22	12.65	11.64	92	58	53	12.14
46	27.02	30.99	115	59	67	28.94
100	57.21	73.17	128	57	73	64.70

 n.d. = not detectable

Validity criteria fulfilled:

yes

Description of key information

EC50 (48h) = 0.43 mg/L for Daphnia magna (OECD 202)

Additional information

Two studies investigating the short-term toxicity of N-methyl-N-(C18-(unsaturated)alkanoyl)glycine (EC 701-177-3) to aquatic invertebrates are available.

The key study was carried out according to OECD guideline 202 and GLP including analytical measurement. Daphnia magna was exposed to the following ten nominal concentrations 0.1, 0.22, 0.46, 1.0, 2.2, 4.6, 10, 22, 46, 100 mg/L under static conditions. These test concentrations were performed in two different tests. The first test was conducted using seven test concentrations (nominal: 1.0, 2.2, 4.6, 10, 22, 46, 100 mg/L). As immobilization (≥ 70%) was observed in all test concentrations, a second test was performed using three additional concentrations (nominal: 0.1, 0.22, 0.46 mg/L). Analytical measurements verified the test item concentration to be 44 – 128% of the initial test concentrations of 1 – 100 mg/L throughout the exposure period, 82 – 363% for the initial test concentrations of 0.1 – 0.46 mf/L. For the test concentrations 0.1 - 0.46 mg/L after 48 h test concentrations of < LOD are detectable. Immobility could be observed in concentrations ≥ 1 mg/L (nominal).The test revealed an EC50 (48h) of 0.43 mg/L (measured).

The supporting study was carried out under non-GLP conditions according to EU Method C.2 and without analytical measurement. Daphnia magna was exposed to ten test concentrations (0.098, 0.195, 0.39, 0.781, 1.56, 3.12, 6.25, 12.5, 25.0, 50.0 mg/L) under static conditions. Immobility of 100% could be observed in concentrations ≥ 0.781 mg/L. An EC50 (48h) of 0.53 mg/L (nominal) is reported.