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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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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:
2016-07-22 till 2017-01-11
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Concentration control analyses were not performed, because a sufficiently sensitive method to determine the test substance in test water up to its saturation limit was not available.
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
Concentration control analyses were not performed
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
2008
Deviations:
yes
Remarks:
Concentration control analyses were not performed
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
The test substance is a mixture of sparingly water soluble components. Therefore test solutions were prepared separately for each test concentration as a water accommodated fraction (WAF) using a liquid-liquid saturator technique. For each test solution, a glass aspirator bottle (2-L) with a bottom side-outlet attached to a stopcock was filled with 2-L test media. Prior to use the test substance was homogenized by shaking the container. Then test substance (considering density = 0. 8166 g/cm³) was pipetted carefully on the water surface. The bottle was closed and the solution was stirred on a magnetic plate for about 3 days. The stirring was slow (approx. 100 rpm) so that no vortex forms. Undissolved test substance was observed at the water surface as oily drops. Yellowish suspended particles were observed in the aqueous phase of the control and saturated test solution (likely dust or otherwise insoluble parts of test medium). The required volume of the WAF (approx. 1 L per test solution) was removed from the bottom port and was clear without visible undissolved substance. The control was treated in the same way. Fresh test solutions were prepared daily. The test vessels were conditioned with the test solution for approx. 24-h prior initiating exposure. The WAF was changed after 24 hours.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna STRAUS 1820
- Source: Institut National de Recherche Chimique Appliquée, France, in 1978. From this date on this clone was cultured and bred continuously in the Ecotoxicology Laboratory of Experimental Toxicology and Ecology, BASF SE, Ludwigshafen Germany.
- Age at test initiation: <24 hours (at least 3rd brood progeny)
- Feeding during test: no

ACCLIMATION
The Daphnia are cultured under the identical conditions as the test including test media (Elendt M4), water quality, and temperature (20 ±1°C).
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
not performed
Hardness:
no data
Test temperature:
19.8 – 20.2 °C
pH:
7.7 – 8.0
Dissolved oxygen:
8.2 - 9.2 mg/L
Conductivity:
550 - 650 μS/cm
Nominal and measured concentrations:
10 - 22 - 46 - 100 - 220 mg/L nominal concentrations based on test substance density without a correction for purity
Details on test conditions:
TEST SYSTEM
- Test vessel: Numbered glass beakers (nominal volume 100 mL)
- Fill volume: 50 mL
- Aeration: no
- No. of organisms per vessel: 5 animals / test vessel
- No. of vessels per concentration: 4 replicates
- No. of vessels per control: 4 replicates
- Biomass loading rate: 0.1 animals/mL
- Renewal of test solution: 24 hours

TEST MEDIUM PARAMETERS
- Preparation of dilution water: A synthetic fresh water (Elendt M4) is used as media for culture and test purposes. For the composition of Elendt M4 medium see OECD 202.
- Total organic carbon: 0.9 mg/L
- Acid capacity up to pH 4.3: 0.84 mmol/L
- Molar ratio Ca:Mg: about 4 : 1
- pH value: 7.9
- Conductivity: 550 - 650 μS/cm
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light : 8 hours darkness
- Light intensity: About 161 – 677 lux at a wave length of 400 - 750 nm

EFFECT PARAMETERS MEASURED :
visually checked for immobilized daphnids after 0, 24 and 48 hours

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: 48 hour EC50 was between 10 and 100 mg/L. Since there was 60% effect at 100 mg/L, concentrations >100 mg/L were included in the definitive test in order to estimate the EC50.
Reference substance (positive control):
yes
Remarks:
sodium chloride, CAS: 7646-14-5, Expiry date: 2018-10-19
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
44.3 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
Remarks on result:
other: Lowest Concentration with 100% Immobile
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
22 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Behavioural abnormalities:
- Observations on body length and weight: no
- Mortality of control: no
- Abnormal responses: no
- Any observations that might cause a difference between measured and nominal values: no

Dissolution behavior:
All of the test solutions were visibly clear and colorless after preparation and remained so after 24 and 48 hours of exposure in the test. No undissolved test substance was visible and there were no other remarkable observations.
Results with reference substance (positive control):
- Results with reference substance valid: yes
The EC50(48h) of the reference substance sodium chloridewas 4.47 g/L (experiment date: 12 Jul 2016). This result is within the range of 3.88 – 7.22 g/L and indicates that the culture of Daphnia magna used in this study is responding normally to toxic stress.
Reported statistics and error estimates:
For the statistical evaluation of the data and calculation of the EC50 the probit method (Finney, 1971) was used and performed with the commercial software "TOXRAT Professional 2.10” (ToxRat Solutions GmbH, Alsdorf, Germany).

Table 1: Number of mobile Daphnia at each observation period

Loading rate [mg/L]

Replicate 1

Replicate 2

Replicate 3

Replicate 4

Sum

Sum

0 h

24 h

48 h

0 h

24 h

48 h

0 h

24 h

48 h

0 h

24 h

48 h

24 h

48 h

0 (control)

5

5

5

5

5

5

5

5

5

5

5

5

20

20

10

5

5

5

5

5

5

5

5

5

5

5

5

20

20

22

5

5

5

5

5

5

5

5

5

5

5

5

20

20

46

5

5

1

5

5

2

5

5

2

5

5

3

20

8

100

5

4

0

5

4

0

5

3

0

5

4

0

15

0

220

5

0

0

5

0

0

5

0

0

5

0

0

0

0

Number of test animals per replicate: 5

Number of test animals per test group: 20

Validity criteria fulfilled:
yes
Conclusions:
The acute immobilisation of the test item to daphnia magna was determined according to OECD guideline 202. The EC50 value after 48 h was determined to be 44.3 mg/L.
Executive summary:

The acute immobilisation of the test item to Daphnia magna (STRAUS) was determined according to OECD Guideline 202 / EEC Directive 92/69/EEC Method C.2. The study was conducted under static-renewal conditions over a duration of 48 hours. 20 test organisms were exposed to each test concentration and control. The nominal test item concentrations were: 10 - 22 - 46 - 100 - 200 mg/L. Four replicates (with five Daphnids) for each test concentration were used.

The EC50 value of the reference item at 4.47 g/L after 48 h was within the prescribed concentration range of 3.88 – 7.22 g/L. Water quality parameters of pH-value and dissolved oxygen concentration measured at 0 and 48 h were determined to be within the acceptable limits.The validity criteria of the test guideline were fulfilled. The EC50 value was determined to be 44.3 mg/L after 48 h.

Description of key information

Key study: The acute immobilisation of the test item to daphnia magna was determined according to OECD guideline 202. The EC50 value after 48 h was determined to be 44.3 mg/L.


Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
44.3 mg/L

Additional information

Key study: The acute immobilisation of the test item to Daphnia magna (STRAUS) was determined according to OECD Guideline 202 / EEC Directive 92/69/EEC Method C.2. The study was conducted under static-renewal conditions over a duration of 48 hours. 20 test organisms were exposed to each test concentration and control. The nominal test item concentrations were: 10 - 22 - 46 - 100 - 200 mg/L. Four replicates (with five Daphnids) for each test concentration were used.

The EC50 value of the reference item at 4.47 g/L after 48 h was within the prescribed concentration range of 3.88 – 7.22 g/L. Water quality parameters of pH-value and dissolved oxygen concentration measured at 0 and 48 h were determined to be within the acceptable limits.The validity criteria of the test guideline were fulfilled. The EC50 value was determined to be 44.3 mg/L after 48 h.