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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 Dec - 16 Dec 2016
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
according to guideline
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
Details on sampling:
- Concentrations: all
- Sampling method: sampling tool pleace at the start and and after 24 h from fresh and after 48 hours from the aged test solutions. 500 μL samples were taken and stabilized with 500 μL methanol.
- Sample storage conditions before analysis: All samples were stored deep frozen.
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test item for preparing the stock solution was weighed and transferred to a volumetric flask. Test medium was added to the bench mark and the solution was shaken and treated for 4 minutes with ultrasonification. Test solutions were prepared by dilutions of the stock solution with the test medium.
The preparation procedure was repeated after 24 hours to obtain the new test solutions.
Test organisms (species):
Daphnia magna
Details on test organisms:
- Common name: water flee
- Strain/clone: Daphnia magna Straus, Clone V
- Source: in house laboratory bred culture. Daphnids originally purchased in a healthy condition from the Federal Environment Agency in Berlin/Germany.
- Feeding during test: none

Test type:
Water media type:
Limit test:
Total exposure duration:
48 h
Post exposure observation period:
13°dH (232 mg/L as CaCO3)
Test temperature:
18.8 – 21.4 °C
control: 7.48 – 7.99
Dissolved oxygen:
9.0 ± 0.1 mg/L
Nominal and measured concentrations:
Nominal: 1.07, 2.35, 5.17, 11.4, 25.0 mg/L
Measured concentrations were between 91% and 103 % of nominal. In the aged samples the concentration of the test substance was 54% of nominal.
Details on test conditions:
- Test vessel: Glass vessel of 100 mLvolume filled up with 50 mL test solution
- Type (delete if not applicable): closed (covered with a glass plate)
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): each 24 hours
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

- Source/preparation of dilution water:
- Test medium: Elendt M4
- Intervals of water quality measurement: each 24 hours

- Adjustment of pH: no
- Photoperiod: 16 hours photoperiod /8 hours darkness daily
- Light intensity: not stated

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mobility after 24 hours and 48 hours

no data
Reference substance (positive control):
potassium dichromate
48 h
Dose descriptor:
Effect conc.:
6.12 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
48 h
Dose descriptor:
Effect conc.:
>= 3.305 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
Details on results:
- Behavioural abnormalities:
- Mortality of control: none
Results with reference substance (positive control):
EC50 (24 h) < 2 mg/L

Immobilization [%] of daphnids after 24 and 48 hours:

Concentration (nominal) of test substance [mg/L] 24 hours 48 hours
0 0 0
1.07 15 20
2.35 5 10
5.17 0 20
11.4 20 90
25 50 100

Immobilization [%] of daphnids by the reference substance after 24 and 48 hours:

Concentration of reference substance [mg/L] 24 hours 48 hours
1 10 100
2 100 100

Description of key information

 48h EC50 (D. magna, mortality) = 3.305 mg/L (measured, OECD 202) 

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
3.305 mg/L

Additional information

The acute toxicity study with aquatic invertebrates (Unger, 2017) was performed accordign to OECD 202 guideline and GLP principles. D. magna were exposed in semi-static conditions to C14-16 AS Na (CAS 91648-54-3) to five test concentrations: 1.07, 2.35, 5.17, 11.4 and 25.0 mg/L (nominal). Four replicates of each test concentration were used and five daphnids were exposed per test concentration. Measured concentrations were between 91% and 103 % of nominal. In the aged samples the concentration of the test substance was 54% of nominal.

The 48-hr EC50 value based on mobility was 6.12 mg/L based on the nominal concentration and equal or higher than 3.305 mg/L for the measured concentration.