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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
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
according to guideline
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
according to guideline
EU Method C.2 (Acute Toxicity for Daphnia)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
Details on sampling:
At the start of the test, four samples (50 mL) were taken from freshly prepared control and test media.
After 48 hours, four samples (50 mL) of control and test media were taken from the pooled media in replicate vessels.
Samples were stored in a freezer until analysis.
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: water accommodated fractions (WAFs)
100 mg test material was dispersed in 1L dilution medium (Elendt M4 medium) in a glass vessel. The contents of the vessel were stirred for approximately 24 hours and then left to stand for approximately 48 hours. An aliquot (800 mL) was then removed mid-vessel to provide the Water Accommodated Fraction (WAF).
Test organisms (species):
Daphnia magna
Details on test organisms:
- Source: in-house culture originating from a strain supplied by the National Institute for Applied Chemical Research (IRCHA), France
- Age at study initiation: < 24 h
- Method of breeding: Stock cultures of Daphnia magna were maintained in glass vessels containing approximately 0.5 to 0.8 litres of Elendt M4 culture medium in a temperature-controlled laboratory at nominally 20 ± 2°C. A photoperiod of 16 hours light : 8 hours dark was maintained, with periods of subdued lighting at the beginning and end of each light phase. The culture medium was renewed three times each week.
Cultures were fed daily with a suspension of the unicellular green algae, Pseudokirchneriella subcapitata, to provide nominally 0.1 to 0.2 mg carbon per daphnid, per day, except during the initial three days when a slightly lower ration was given. Culture conditions ensure that the stock animals reproduce by parthenogenesis.
The day before the start of the study, all juvenile Daphnia were removed from the laboratory cultures. The following morning, juveniles produced by the gravid (egg-bearing) adult Daphnia were removed from the culture vessels and held in a separate holding vessel.
- Feeding during test: none

- Acclimation period: not relevant, conditions were the same for maintenance and testing
Test type:
Water media type:
Limit test:
Total exposure duration:
48 h
260-270 mg/L as CaCO3
Test temperature:
19.5-20 °C
Dissolved oxygen:
96-106 % air saturation value
Nominal and measured concentrations:
nominal loading rate: 100 mg test material/L
mean measured TOC levels at test start: 2.26 mgC/L
mean measured TOC levels at test end: 2.23 mgC/L
geometric mean value: 2.24 mgC/L
Details on test conditions:
- Test vessel: glass dishes containing approx. 120 mL of medium, covered leaving no headspace
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 24 mL medium per organism

- Source/preparation of dilution water: Elendt M4 medium (according to guideline), prepared in deionised, reverse osmosis water
- Total organic carbon: 1.4 mg C/L
- Alkalinity: 38-50 mg/L as CaCO3
- Ca/Mg ratio: 3.7:1
- Culture medium different from test medium: no

- Adjustment of pH: no
- Photoperiod: 16 hours light : 8 hours dark with periods of subdued lighting at the beginning and end of each light phase

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : numbers of mobile, immobile and floating Daphnia counted approx. 24 and 48 hours after the start of the study

- Test concentrations: nominal loading rates of 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: no significant immobility after 48 hours.
Reference substance (positive control):
potassium dichromate
Key result
48 h
Dose descriptor:
Effect conc.:
> 100 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: results are expressed in terms of loading rates
Key result
48 h
Dose descriptor:
Effect conc.:
>= 100 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: results are expressed in terms of loading rates
Details on results:
- Immobilised Daphnia: none
- Behavioural abnormalities: none
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none

The test medium was a colourless dispersion.
Results with reference substance (positive control):
- 48-hour EC50: 1.3 mg/L (within the range typically obtained in this laboratory)
Reported statistics and error estimates:
The “no observed effect loading rate” (NOELR) was derived by direct inspection of the data on the immobility of the animals. An incidence rate of more than 10% was considered to be significant.
Validity criteria fulfilled:
In the control, no Daphnia have been immobilised or trapped at the surface of the water. The dissolved oxygen concentration at the end of the test was ≥60% of the air saturation value at the temperature used.

Description of key information

48-h EC50 (based on loading rates) >100 mg/L for Daphnia magna (OECD TG 202, EU C.2)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
100 mg/L

Additional information

The EC50/LC50 for the test material is much higher than its water solubility.