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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1988-04-06 - 1988-04-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The experiment was conducted according to EU Method C.2, meeting scientific principles. No information about GLP compliance is reported. Not all details are described in test report; however, available data indicate that the study was well-performed.
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material: No surrogate or analogue material was used.
Analytical monitoring:
no
Details on sampling:
Samples were taken after 3, 6, 24 and 48 h after onset of the experiment.
Vehicle:
no
Details on test solutions:
The concentration of the test substance in the stock preparation was 500 mg/L. A dilution series was prepared with dilution water. 10 mL of the specific dilution was added per test vessel.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Strain: Straus
- Source: Institut National de Recherche Chimique Appliquée, France
- Age of animals: 2 - 24 h

CULTURE CONDITIONS
- Location: Laboratory of Environmental Analytics and Ecology of BASF AG Ludwigshafen, Germany since 1978
- Total water hardness: 2.7 +/- 0.5 mmol/L; Mole ratio Ca:Mg: about 4:1; Oxygen content > 2 mg/L
- Temperature: 18.85 - 20.85 °C
- Feed: Brewer´s yeast when the water is changed and washed green alga culture once daily
- Illumination: 16 h light, 8 h dark
- Illuminance: about 5 microEinstein (m*m*s) in the wavelength range of 400 - 750 nm
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
No post exposure observation period described.
Hardness:
3.0 mmol/L (start of the experiment)
Test temperature:
18.75 °C (291.9 K; start of the experiment)
pH:
0 mg/L: 7.94 (0h), 8.00 (48h)
31.25 mg/L: 7.97 (0h), 7.64 (48h)
62.50 mg/L: 8.02 (0h), 7.55 (48h)
125 mg/L: 8.04 (0h), 7.23 (48h)
250 mg/L: 8.06 (0h), 7.17 (48h)
500 mg/L: 8.01 (0h), 7.11 (48h)
Dissolved oxygen:
0 mg/L: 9.40 mgO2/L (0h), 8.54 mgO2/L (48h)
31.25 mg/L: 9.28 mgO2/L (0h), 6.70 mgO2/L (48h)
62.50 mg/L: 9.15 mgO2/L (0h), 6.32 mgO2/L (48h)
125 mg/L: 9.06 mgO2/L (0h), 3.68 mgO2/L (48h)
250 mg/L: 9.03 mgO2/L (0h), 3.50 mgO2/L (48h)
500 mg/L: 8.99 mgO2/L (0h), 2.64 mgO2/L (48h)
Salinity:
Not applicable.
Nominal and measured concentrations:
0, 31.25, 62.50, 125, 250 and 500 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Centrifuge glasses with flattened bottoms
- Test volume: 10 mL
- Aeration: yes, with oil-free air
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 2 mL

TEST MEDIUM / WATER PARAMETERS
- Source/preparation: Tap water was used as test water, which was passed over a filter candle of 6 µm and over an activated carbon filter to remove any chlorine present. Analytical-grade H2SO4 was added to reduce the Ks pH 4.3. Distilled water was added to reduce the total hardness.
- Mole ratio Ca:Mg: 4:1
- Mole ratio Na:K: 10:1
- Conductivity: 550 - 650 µSiemens/cm

OTHER TEST CONDITIONS
- Photoperiod: 16h light, 8h dark

RANGE FINDING STUDY
None
Reference substance (positive control):
not required
Remarks:
Not mandatory under the applied guideline.
Duration:
24 h
Dose descriptor:
EC0
Effect conc.:
250 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 500 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC100
Effect conc.:
>= 500 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC0
Effect conc.:
125 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
500 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
> 500 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
EC0(3h): 500 mg/L, EC50(3h) >= 500 mg/L, EC100(3h) > 500 mg/L
EC0(6h): 500 mg/L, EC50(6h) >= 500 mg/L, EC100(6h) > 500 mg/L
Results with reference substance (positive control):
Not applicable.
Reported statistics and error estimates:
No statistics or error estimates reported.

Table 1. Test animals able to swim

Concentration
 [mg/L]

Test animals able to swim after

0h

3h

6h

24h

48h

0.00

20

20

20

20

20

31.25

20

20

20

20

20

62.50

20

20

20

20

20

125.00

20

20

20

19

16

250.00

20

20

20

14

11

500.00

20

20

20

20

20

Validity criteria fulfilled:
yes
Remarks:
Scientific criteria for acute toxicity testing to aquatic invertebrates seemed to be fulfilled.
Conclusions:
The study report describes a valid study, conducted according to EU Method C.2. No information about GLP compliance available. Available data confirm that the experiment was well-performed. The test substance was not toxic to Daphnia magna up to a concentration of 500 mg/L during an exposure duration of 48 h.
Executive summary:

The acute toxicity of Choline chloride towards the aquatic invertebrate Daphnia magna was investigated according to EU Method C.2. (BASF AG, 1988). Test concentrations were chosen as followed: 0 (control), 31.25, 62.50, 125, 250 and 500 mg/L. Four replicates of each concentration were included. The mobility and the corresponding EC0, EC50 and EC100 values were determined after 3, 6, 24 and 48 hours. The highest concentration tested at which an effect

10 % occurred was referred to as EC0. Physico-chemical measurements (temperature, pH and dissolved oxygen content) were measured at the beginning and termination of the experiment. At the start, the temperature was recorded as 18.75 °C and the pH 7.8. The exposure duration was 48 h. After this time, the final results were reported as: EC0(48h): 125 mg/L, EC50(48h): 500 mg/L and EC100(48h) >= 500 mg/L. No analytical verification and no statistical methods were implemented in the experiment.

Description of key information

No toxicity observed even at highest concentration level tested.
Daphnia magna_EU Method C.2: EC0(48h): 125 mg/L, EC50(48h): 500 mg/L, EC100(48h) 500 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
500 mg/L

Additional information

The acute toxicity of Choline chloride towards the aquatic invertebrate Daphnia magna was investigated according to EU Method C.2. (BASF AG, 1988). Test concentrations were chosen as followed: 0 (control), 31.25, 62.50, 125, 250 and 500 mg/L. Four replicates of each concentration were included. The mobility and the corresponding EC0, EC50 and EC100 values were determined after 3, 6, 24 and 48 hours. The highest concentration tested at which an effect

10 % occurred was referred to as EC0. Physico-chemical measurements (temperature, pH and dissolved oxygen content) were measured at the beginning and termination of the experiment. At the start, the temperature was recorded as 18.75 °C and the pH 7.8. The exposure duration was 48 h. Results were reported as: EC0(48h): 125 mg/L, EC50(48h) >= 500 mg/L and EC100(48h) >= 500 mg/L. No analytical verification and no statistical methods were implemented in the experiment.

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