N-[3-({[2,2-dimethyl-3-(morpholin-4-yl)propylidene]amino}methyl)-3,5,5-trimethylcyclohexyl]-2,2-dimethyl-3-(morpholin-4-yl)propan-1-imine

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 2012-05-16 to 2012-05-18

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

For determination of the test item concentration, samples were taken from the testing concentrations and the control at the start and at the end of each test.

Vehicle:

no

Details on test solutions:

A stock solution of 100 mg/L (nominal) was first prepared by dissolving an amount of 0.10 g test item in 1000 mL ISO medium using intensive shaking (until all of the test item dissolved). The test solutions of the chosen test concentrations were prepared by appropriate dilution of this stock solution.
The test solutions were prepared immediately before introduction of daphnids. Preparation of test solution was performed by a same way at each water renewal period.

Test organisms (species):

Daphnia magna

Details on test organisms:

- Species: Daphnia magna (Straus)
- Age at test start: Less than 24 hours old
- Sex: Female
- Origin: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection) 2100 Gödöllő, Kotlán S. u. 3., Hungary
- Breeding conditions: Daphnia were bred in the Laboratory of TOXI-COOP ZRT. under similar temperature and light conditions as in the test, and in reconstituted water of a similar quality regarding to pH, components of the main ions and total hardness as the test water used in the test. The test animals were fed with centrifuged green alga suspension.
- Acclimatisation: Test animals were bred under similar conditions as that used during the exposure period, therefore, additional acclimatisation before the test was not necessary.
- Number of animals: Twenty animals at each group, divided into 4 replicates.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

249 mg/L (as CaCO3)

Test temperature:

19.5 – 20.2 °C in the test vessels;
within the climate chamber 19.1 – 20.6 °C

pH:

7.91 – 9.23

Dissolved oxygen:

7.65 – 8.33 mg/L

Salinity:

N/A

Nominal and measured concentrations:

nominal concentrations: 9.5, 17.1, 30.9, 55.6 and 100.0 mg/L
measured concentrations: 9.97, 19.03, 31.57, 57.48, 103.33 mg/L (102 and 114 % of the nominal concentration)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers of approximately 40 mL test medium
- No. of organisms per vessel: Twenty animals at each group, divided into 4 replicates.


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light (artificial illumination) and 8 hours darkness

EFFECT PARAMETERS MEASURED:
EC 50: concentration estimated to immobilise 50 per cent of the daphnids within a stated exposure period
EC100: the lowest test concentration at which all test animals are immobile.
NOEC (48 h, No Observed Effect Concentration) the highest test concentration at which no significant immobilisation or abnormal behaviour is observed relative to the control.
LOEC: (Lowest Observed Effect Concentration) the lowest test concentration at which a significant immobilisation or abnormal behaviour is observed relative to the control.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

40.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % conf. limits (71.1 – 94.2 mg/L)

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

17.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

30.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- The immobilization was 80 per cent at the highest tested concentration (100 mg/L) after 24 hours of exposure.
- A single daphnid was observed as immobile at the concentration of 55.6 mg/L.
- At the lower concentrations and in the control no immobility was noticed.
- The 24-h EC50 was calculated to be 81.7 mg/L (95 % confidence limits: 71.1 – 94.2 mg/L) using Probit analysis by SPSS PC+.
- After 48 hours in the concentrations of 9.5 and 17.1 mg/L a statistically not significant immobility was noticed.
- The immobility was statistically significant at the concentration range of 30.9 – 100 mg/L.
- The NOEC was determined to be 17.1 mg/L and the LOEC was determined as 100.0 mg/L.
- The 48-h EC50 was calculated to be 40.2 mg/L (95 % confidence limits: 33.0 – 49.2 mg/L) using Probit analysis by SPSS PC+ software.
- All daphnids were observed as immobile at the concentration of 100 mg/L; correspondingly the 48-h EC100.

Results with reference substance (positive control):

The date of the last study (Study Code: 392.441.3446) with reference item Potassium dichromate was: 22 – 23 March 2012.
The 24-h EC50 value of Potassium dichromate (K2Cr2O7) was determined to be 0.97 mg/L.
This 24-h EC50 value is in accordance with the accepted 0.6 – 2.1 mg/L range for Daphnia magna.

Reported statistics and error estimates:

Statistically not significant immobility (Bonferroni t-Test (a= 0.05)) was noticed in the concentrations of 9.5 and 17.1 mg/L. The immobility was statistically significant (Bonferroni t-Test (a= 0.05)) at the concentration range of 30.9 – 100 mg/L. Accordingly, the NOEC was determined to be 17.1 mg/L and the LOEC was determined as 100.0 mg/L.
The 48-h EC50 was calculated to be 40.2 mg/L (95 % confidence limits: 33.0 – 49.2 mg/L) using Probit analysis by SPSS PC+ software.
All daphnids were observed as immobile at the concentration of 100 mg/L; correspondingly, the 48-h EC100 was determined to be 100 mg/L.

Validity criteria fulfilled:

yes

Conclusions:

In this 48-hour acute toxicity test with Daphnia magna the 48 h EC50 value was determined as 40.2 mg/L, and the 48 h NOEC was determined to be 17.1 mg/L. The results are based on the nominal test item concentrations.

Executive summary:

An acute toxicity test on Daphnia magna over an exposure period of 48 hours in a semi-static system was assessed according to OECD Guideline 202 and EU method C.2. The purpose of this study was to evaluate the influence of the test item Sika Hardener MI on the mobility respectively survival of Daphnia magna. Young Daphnia were exposed using a nominal range of concentration of 9.5, 17.1, 30.9, 55.6 and 100.0 mg/L under semi-static conditions. The analytically measured concentrations of the test item were within ± 20 % of the nominal during the experiment. Therefore, the biological results were based on the nominal concentrations. The 48-h EC50 were determined to be 40.2 mg/L, and the 48-h NOEC was determined to be 17.1 mg/L. The results are based on the measured geometric mean test item concentrations.

Description of key information

In a 48-hour acute toxicity test with Daphnia magna the 48-h EC50 value was determined to be 40.2 mg/L, and the 48-h NOEC was determined to be 17.1 mg/L. The results are based on the nominal test item concentrations.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

40.2 mg/L

Additional information

An acute toxicity test on Daphnia magna over an exposure period of 48 hours in a semi-static system was assessed according to OECD Guideline 202 and EU method C.2. The purpose of this study was to evaluate the influence of the test item Sika Hardener MI on the mobility respectively survival of Daphnia magna. Young Daphnia were exposed using a nominal range of concentration of 9.5, 17.1, 30.9, 55.6 and 100.0 mg/L under semi-static conditions. The analytically measured concentrations of the test item were within ± 20 % of the nominal during the experiment. Therefore, the biological results were based on the nominal concentrations. The 48-h EC50 were determined to be 40.2 mg/L, and the 48-h NOEC was determined to be 17.1 mg/L. The results are based on the measured geometric mean test item concentrations.