1,3,3-trimethyl-N-(2-methylpropylidene)-5-[(2-methylpropylidene)amino]cyclohexanemethylamine

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Sampling method: samples taken from additional beakers without daphnia, kept under the same experimental conditions
- Sample storage conditions before analysis: samples were taken, immediately transferred to the analytical laboratory and analysed on the same day.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test substance concentrations were individually prepared in M4 medium, stirred for 24 h, and afterwards the preparations were filtered .
- Controls: control preparations without test substance but treated the same way.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna, water flea
- Source: Landesumweltamt NRW, Düsseldorf, Germany
- Age at study initiation (mean and range, SD): < 24 h
- Method of breeding: The daphnids were held in beakers with a nominal capacity of 2000 mL. Each beaker was filled with approximately 1400 mL M4 medium (Elendt-Schneider). Between 15 and 20 adult organisms were held in one beaker in order to get a high reproduction rate and viable offspring. The breeding beakers were kept at an additional artificial light-dark cycle of 16 to 8 hours. Breeding beakers were placed in a temperature controlled laboratory at 20 +/- 2°C. The breeding medium was renewed three times a week.
- Feeding during test: no feeding during experimental period


ACCLIMATION
- Acclimation period: culture of daphnids permanently kept at the test facility
- Acclimation conditions (same as test or not): no acclimation necessary
- Type and amount of food: singel cell green algae (Desmodesmus subspicatus)
- Feeding frequency: daily

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

2.502 mmol/L (total hardness)

Test temperature:

20.7 - 22.0°C

pH:

7.95 - 8.28

Dissolved oxygen:

5.5 - 6.2 mg/L

Nominal and measured concentrations:

nominal: 3.75 mg/L, 7.5 mg/L, 15 mg/L, 30 mg/L, 60 mg/L
measured (DOC at 48 h): 20.5 mg/L (nominal 30 mg/L), 31.4 mg/L (nominal 60 mg/L)

Details on test conditions:

TEST SYSTEM
- Type: open
- Material, size, headspace, fill volume: glas test beakers (50 mL), filled with 30 mL
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 5
- No. of vessels per control (replicates): 5
- Biomass loading rate: 1 daphnid per 6 mL test solution

TEST MEDIUM / WATER PARAMETERS
- Total organic carbon: 0 mg/L
- Alkalinity: 0.936 mmol/L
- Conductivity: 588 µS/cm
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 16 h / 8 h light / dark cycle
- Light intensity: 700 +/- 50 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobility

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Results of range finding study used to determine the conditions for the definitive study

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

22.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 17.0-30.2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

14.7 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Remarks:

(DOC)

Basis for effect:

mobility

Remarks on result:

other: 4.9-20.2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

7.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: no immobility observed in the control group
- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

- Results with reference substance valid? yes, test was conducted on 22 February 2011 (24 h)
- Measured parameter: immobility
- EC50/LC50: EC50 after 24 h = 1.0 mg/L; in the required range of 0.6 - 2.1 mg/L

Reported statistics and error estimates:

EC values calculated with ToxRat Professional (ToxRat Solutions GmbH, Alsdorf, Germany)

Analytical data: see attachment

Effect values:

EC value	Effective concentration after 48 hours [mg/L]
	nominal
EC50	22.2
95 % Confidence interval	17.0 – 30.2
NOEC	7.5

 

 

Vessel No.		Concentration of test substance (mg/L)		Immobilised daphnids per test vessel
				24 h					48 h
1 - 5		0 (Control)		0	0	0	0	0	0	0	0	0	0
6 - 10		3.75		0	0	0	0	0	0	0	0	0	0
11 - 15		7.5		0	0	0	0	0	2	0	0	0	2
16 - 20		15		3	2	2	2	2	3	2	2	2	2
21 - 25		30		0	2	3	2	4	2	2	3	3	5
26 - 30		60		3	3	2	3	2	4	4	4	4	4

Validity criteria fulfilled:

yes

Conclusions:

The EC50 (48 h), based on nominal concentrations was determined as 22.2 mg/L indicating that the test substance may be harmful to aquatic invertebrates. Based on DOC, the EC50 (48 h) was 14.7 mg/L.

Executive summary:

The test substance was tested in the Acute Daphnia Toxicity Test according to the OECD Guideline for the Testing of Chemicals No. 202 (April 2004) using Daphnia magna (Straus) as test organism. The experiment was carried out as a static test, i.e. the test substance preparations were not exchanged for the whole exposure period of 48 hours. The test was performed with test substance preparations of nominal 3.75 mg/L, 7.5 mg/L, 15 mg/L, 30 mg/L, 60 mg test substance/L M4 medium. The EC₅₀value after 48 hours of exposure was 22.2 mg/L and the NOEC was 7.5 mg/L, based on nominal concentrations. Based on measured DOC values, the EC₅₀value was 14.7 mg/L.

Description of key information

The test substance was tested in the Acute Daphnia Toxicity Test according to the OECD Guideline for the Testing of Chemicals No. 202 (April 2004) using Daphnia magna (Straus) as test organism. The experiment was carried out as a static test, i.e. the test substance preparations were not exchanged for the whole exposure period of 48 hours. The test was performed with test substance preparations of nominal 3.75 mg/L, 7.5 mg/L, 15 mg/L, 30 mg/L, 60 mg test substance/L M4 medium. The EC50value after 48 hours of exposure was 22.2 mg/L and the NOEC was 7.5 mg/L, based on nominal concentrations. Based on measured DOC values, the EC50 value was 14.7 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

14.7 mg/L