1,6-Hexanediamine, N1,N6-bis[2,2-dimethyl-3-(4-morpholinyl)propylidene]-

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 2012-05-08 to 2012-05-10

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:

,updated 13th April 2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

Nr. 440/2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(1996)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Batch no.: 10191-AKm
- Purity. 100.5 % (aldimine titration)
- Expiry date: 2013-06-16

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 renewal period.

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:

The reconstituted water (ISO medium) had an approximate theoretical total hardness of 249 mg/L (as CaCO3).

Test temperature:

The test temperature was in the range of 19.3 – 19.9 °C measured in the test vessels during the test. The additionally measured ambient temperature within the climate chamber was between 19.0 – 20.1 °C.

pH:

The pH of the test solution was not adjusted and did not vary by more than 1.5 units in any one test. The pH was in the range of 7.61 – 8.95 during the test.

Dissolved oxygen:

The dissolved oxygen concentration was in the range of 7.91 – 8.54 mg/L during the test.

Nominal and measured concentrations:

Based on the results of the preliminary range-finding test, five test concentrations in a geometric series with a separation factor of 1.5 and one control was used in the main test.
The following nominal concentrations were tested: 19.8, 29.6, 44.4, 66.7 and 100.0 mg/L.
The corresponding measured geometric mean test item concentrations were: 19.4, 29.2, 44.0, 63.3 and 98.5 mg/L.
All reported biological results are related to the geometric mean of the measured test item concentrations.

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 h light : 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.

TEST CONCENTRATIONS
- Range finding study: A non-GLP preliminary range-finding tests was performed. In the first range finding test, five widely spaced concentrations (0.01 - 100 mg/L).

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

81 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% conf. limits (72.6 - 91.6)

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 98.5 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

44 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

63.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Immobility was not observed in the control group and in whole tested concentration range (19.4 – 98.5 mg/L) after 24 hours of exposure. The 24-h EC50 was higher than 98.5 mg/L. After 48-h immobility was not observed in the control and in the concentration range of 19.4 – 44.0 mg/L.
- The NOEC was determined to be 44.0 mg/L and the LOEC was determined as 63.3 mg/L. The 48-h EC50 was calculated to be 81.0 mg/L (95 % confidence limits: 72.6 – 91.6 mg/L) .
- The 48-h EC100 value was determined to be higher than 98.5 mg/L.
- Statistically not significant (15 %) immobility was detected at 63.3 mg/L and statistically significant immobility (80 %) was noticed at the highest tested concentration of 98.5 mg/L

Results with reference substance (positive control):

The 24h EC50 value of Potassium dichromate (K2Cr2O7) was determined to be 0.97 mg/L.
This 24-hour EC50 value is in accordance with the accepted 0.6 – 2.1 mg/L range for Daphnia magna.

Reported statistics and error estimates:

EC50 was calculated by Probit analysis.

Validity criteria fulfilled:

yes

Conclusions:

In this 48-hour acute toxicity test with Daphnia magna the 48-h EC50 value was determined as 81.0 mg/L, and the 48-h NOEC was determined to be 44.0 mg/L. The results are based on the measured geometric mean 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 MH on the mobility respectively survival of Daphnia magna. Young Daphnia were exposed using a nominal range of concentration of 19.8, 29.6, 44.4, 66.7 and 100.0 mg/L under semi-static conditions. The measured concentrations deviated more than 20 % from the nominal at some cases therefore the geometric mean of the measured concentrations were calculated in order to determine exposure concentrations. The corresponding calculated geometric mean concentrations were the followings:19.4, 29.2, 44.0, 63.3 and 98.5 mg/L. The 48 -h EC50 was determined to be 81.0 mg/L, and the 48-h NOEC was determined to be 44.0 mg/L. The results are based on the measured geometric mean test item concentrations.

Description of key information

In this 48-hour acute toxicity test with Daphnia magna the 48-h EC50 value was determined as 81.0 mg/L, and the 48 -h NOEC was determined to be 44.0 mg/L. The results are based on the measured geometric mean test item concentrations.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

81 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 MH on the mobility respectively survival of Daphnia magna. Young Daphnia were exposed using a nominal range of concentration of 19.8, 29.6, 44.4, 66.7 and 100.0 mg/L under semi-static conditions. The measured concentrations deviated more than 20 % from the nominal at some cases therefore the geometric mean of the measured concentrations were calculated in order to determine exposure concentrations. The corresponding calculated geometric mean concentrations were the followings:19.4, 29.2, 44.0, 63.3 and 98.5 mg/L. The 48 -h EC50 was determined to be 81.0 mg/L, and the 48 h NOEC was determined to be 44.0 mg/L. The results are based on the measured geometric mean test item concentrations.