2-Propenenitrile, reaction products with 2,2,4(or 2,4,4)-trimethyl-1,6-hexanediamine

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:

from 1993-05-04 to 1993-06-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

EG 84/449, 1989

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

Part 1, 1984

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: DIN 38412 Part 1

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

A stock solution was prepared with concentration of 1 g/L in deionised water.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Clone: 5
- Strain: Straus
- Source: in-house culture.
The culture is kept in M4 Medium in 1 L beakers. Every 2-3 days the medium is changed and the young daphnids are separated.
- Feeding of the culture
- Food type: Scenedesmus subspicatus
- Amount: calculated in order for the animals to consume the whole amount
- Frequency: daily

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 °C

pH:

7.8 - 8.3

Dissolved oxygen:

8.3 - 8.7 mg/L

Nominal and measured concentrations:

Nominal concentrations: 0, 0.8, 1.5, 3, 6, 12, 25, 50, 100 mg/L
No measured concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: round bottom glasses
- Material, size, headspace, fill volume: glass, 10 mL
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Feeding during the test: no

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium, prepared according to OECD 202
- Composition of test medium:
CaCl2x2H2O - 294 mg/L
MgSO4x7H2O - 123 mg/L
NaHCO3 - 63 mg/L
KCl - 5.5 mg/L
- Na/K ratio: 10:1
- Ca/mg ratio: 2.5 mmol; 4:1
- Culture medium different from test medium: yes

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: continuous darkness

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

31.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI: 22.9 - 43.9 mg/L

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

19.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI: 14.1 - 27.7 mg/L

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

After 48 h the highest concentration at which no immobilisation (<= 10 %) was observed, was determined to be 3 mg/L.
After 48 h the highest concentration at which 100 % immobilisation was observed, was determined to be 100 mg/L.
All biological effects are based on the nominal concentrations.

Results with reference substance (positive control):

Potassium dichromate was tested as reference substance. After 24 h exposure at concentration level of 0.9 mg/L 30 % immobilisation was observed. 100 % immobilisation was observed at 1.9 mg/L.

Table 1. Individual results on mobility after 24 and 48 h

Concentration (mg/L)	24 h			48 h
	Animals		%	Animals		%
	Mobile	Immobile	Immobile	Mobile	Immobile	Immobile
Control	19	1	5	19	1	5
0.8#	19	0	0	18	1	5
1.5*	21	0	0	21	0	0
3.0	20	0	0	20	0	0
6.0	19	0	0	17	2	11
12.0	16	4	20	11	9	45
25.0	15	5	25	12	8	40
50.0	6	14	70	3	17	85
100.0	0	20	100	0	20	100

# 21 daphnids were counted at the beginning of the test.

* 19 daphnids were counted at the beginning of the test.

Table 2. EC50 values calculated after 24 and 48 h exposure

Time (h)	EC50 (mg/L)	95 % confident intervals (mg/L)
24	31.7	22.9 – 43.9
48	19.7	14.1 – 27.7

Validity criteria fulfilled:

yes

Conclusions:

The EC50 of the test item on Daphnia magna was determined after 48 h exposure to be 19.7 mg/L. The NOEC was determined to be 3 mg/L.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item were determined according to OECD 202, Regulation (EC) No. 440/2008 Method C.2 and DIN 38412 Part 1. The study was conducted in a static system over a period of 48 hours with a concentration series of 0.8, 1.5, 3, 6, 12, 25, 50 and 100 mg/L. 20 daphnids per test concentration and control divided in 5 replicates with 4 animals each were tested. The test item concentration was not determined analytically, therefore, the biological endpoints were based on the nominal concentration. After 48 h exposure significant effects on the daphnids were observed. The NOEC was determined to be 3 mg/L. The concentration at which 100 % immobilization was observed was determined to be 100 mg/L. The EC50 after 48 h exposure was determined to be 19.7 mg/L. Significant effects of the test item on Daphnia magna were observed under the test conditions.

Description of key information

The EC50 of the test item on Daphnia magna was determined after 48 h exposure to be 19.7 mg/L. The NOEC was determined to be 3 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

19.7 mg/L

Additional information

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item were determined according to OECD 202, Regulation (EC) No. 440/2008 Method C.2 and DIN 38412 Part 1. The study was conducted in a static system over a period of 48 hours with a concentration series of 0.8, 1.5, 3, 6, 12, 25, 50 and 100 mg/L. 20 daphnids per test concentration and control divided in 5 replicates with 4 animals each were tested. The test item concentration was not determined analytically, therefore, the biological endpoints were based on the nominal concentration. After 48 h exposure significant effects on the daphnids were observed. The NOEC was determined to be 3 mg/L. The concentration at which 100 % immobilization was observed was determined to be 100 mg/L. The EC50 after 48 h exposure was determined to be 19.7 mg/L. Significant effects of the test item on Daphnia magna were observed under the test conditions.