1,3-Cyclohexanedimethanamine, N1,N3-bis(2-methylpropylidene)-

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:

2010-11-12 to 2011-10-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study, in accordance to OECD, EU guidelines

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

updated 13th April 2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

dated May 30, 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Sampling preparation:
Three replicate samples were taken from each test concentration and from the control at the start and at the end of the study.
Representative samples (in duplicate series) were taken at the start and at the end of the experiment. One series are stored frozen in the testing laboratory, one series were analysed immediately after sampling.

- Storage of the Samples: The second series of samples were stored in a refrigerator immediately after sampling, and were kept stored until delivery of the final report to enable additional analyses on request of the sponsor. After delivery of the final report, all samples will be discarded.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Formulation: At the preparation of the test item stock solution 50 mg test item was weighted and emulsified in 500 mL ISO Medium, thereafter sonicated for 30 minutes in ultrasonic bath (under cooling). The „stock solution” was centrifuged at 4000 rpm for 10 min. (20 °C) and filtrated using a 0.45 μm filter (VWR Vacuum Filtration System) and further diluted accordingly.
Untreated Control: ISO Medium with daphnids (without test item) was examined in parallel in the experiment.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection) 2100 Gödöllő, Kotlán S. u. 3. Hungary
- Age: Less than 24 hours old
- Sex: female
- Feeding, Aeration: None
- Acclimatization: The Daphnia mothers selected ca. one months prior to the start of the test were maintained in ISO medium under test conditions (temperature, light conditions, quality and content of reconstituted water, pH and total hardness).
The test animals were fed daily with a few mL of centrifuged algae (Pseudokirchneriella subcapitata) suspension.
To avoid the necessity of longer adaptation prior to the test, the water used in the test is similar (or the same) to the culture conditions (water temperature, background colour, light intensities).

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

219.5 mg/L

Test temperature:

21.1 – 21.9 °C in the climate chamber
21.4 – 21.6 °C in the test media

pH:

pH 7.97 to 8.66 at the start of the test
pH 7.91 to 8.19 at the end of the test

Dissolved oxygen:

8. 37 to 8.83 mg/L at test start of the test
8.16 to 8.31 mg/L at test end of the test

Nominal and measured concentrations:

Nominal 6.25; 12.5; 25, 50 and 100 mg/L
Mean measured concentrations with 95 % confidence interval: 5.46 ± 0.062, 13.3 ± 0.15, 27.1 ± 0.28, 57.5 ± 2.05, 113.1 ± 1.63

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers of 50 mL volume with ~40 mL test medium
- No. of organisms per vessel: 20 Daphnia per control(s) and test concentration, divided into four groups of five animals, each group in at least 40 mL test medium

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light : 8 h dark

EFFECT PARAMETERS MEASURED:
NOEC (48 h): No Observed Effect Concentration: highest test concentration at which no significant effect is determined after 48 hours test duration
EC 50: the calculated concentration of the test item which results in a 50 % immobilisation rate
EC 0: highest test concentration without a significant number of immobilised test animals
EC 100: the lowest test concentration at which all test animals are immobile

TEST CONCENTRATIONS
- Range finding study: Two non-GLP preliminary range-finding tests were performed. In the first range-finding test the examined nominal concentrations were: 3.125, 6.25, 12.5, 25, 50 and 100 mg test item/L. In the second range-finding test the same concentration range was investigated.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

68.79 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL: 56.88 - 85.19 mg/L

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

The actual 24-hours EC50 value of the potassium dichromate is: 2.05 mg/L.
This 24-hour EC50 value is in accordance with the accepted 0.6 – 2.1 mg/L range for Daphnia magna.

The analytically determined mean test item concentrations in the analysed test media varied in the range from 86 % to 103 % of the nominal value at the start of the study and from 87 % to 116 % at the end of the study (see the attached analytical report). The measured concentrations remained within the range of ± 20% of nominal during the study, therefore, all reported biological results are related to the nominal concentrations applied in the test.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this acute daphnia toxicity study the observed endpoints for the effect of Incorez 397 were the followings: 48h EC50 value: 68.79 mg test item/L; 48h NOEC value: 25 mg test item/L and 48h LOEC value: 50 mg test item/L. All reported biological results are related to the nominal concentrations.

Executive summary:

An acute toxicity test on Daphnia magna over an exposure period of 48 hours in a static system was assessed according to OECD Guideline 202 and EU method C.2. Five different concentrations of the test item Incorez 397 arranged in a geometric series and an untreated control group was tested in the main experiment. The concentrations were in nominal: 6.25; 12.5; 25, 50 and 100 mg/L. The 48-hour EC50value was determined as 68.79 mg/L (Probit analysis).The 48-hour NOEC was determined to be 25 mg/L, while the 48-hour LOEC was 50 mg test item/L. All reported biological results are related to the nominal concentrations.

Description of key information

Under the conditions of this acute daphnia toxicity study the observed endpoints for the effect of Incorez 397 were the followings: 48h EC50 value: 68.79 mg test item/L; 48h NOEC value: 25 mg test item/L and 48h LOEC value: 50 mg test item/L. All reported biological results are related to the nominal concentrations.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

68.79 mg/L

Additional information

An acute toxicity Test on Daphnia magna over an exposure period of 48 hours in a static system was assessed according to OECD Guideline 202 and EU method C.2. Five different concentrations of the test item Incorez 397 arranged in a geometric series and an untreated control group was tested in the main experiment. The concentrations were in nominal: 6.25; 12.5; 25, 50 and 100 mg/L. The48-hour EC50value was determined as 68.79  mg/L (Probit analysis). The 48-hour NOEC was determined to be 25 mg/L, while the 48-hour LOEC was 50 mg test item/L. All reported biological results are related to the nominal concentrations.