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Ecotoxicological information

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:
from 2000-08-04 to 2000-10-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
1984
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Before analysis the samples had to be prepared and diluted. So the analytical test material concentrations of 1.0 and 1.8 mg/L (nominal concentrations) got below the limit of quantification (5 mg/L). Therefore the three high concentrations were analyzed. The analytical controls were carried out with 3.2, 5.6 and 1 mg/L
- Sampling method: For the analytical measurement a sample from the freshly prepared test medium was taken at the start of the study. For the determination of the maintenance of the test material concentration during the exposure period a sample was taken from the medium at the end of the exposure period.
- Sample storage conditions before analysis: tightly closed, dark, at room temperature
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A stock preparation with a test material concentration of 100 mg/L was prepared. The test medium (reconstituted water and test material) was freshly prepared. Therefore, the calibrated flask with test material and vehicle, reconstituted water, was treated in an ultrasonic device for 1 hour. Subsequently the preparation was aerated, and stirred with a magnetic stirrer for further 23 hours. After 24 hours the formulation was given through a nutsch filter (pore size >10 - < 16 µm). The filtrate was used for the stock preparation. The stock preparation was diluted with reconstituted water to the different test medium concentrations.
- Controls: Blank control performed with the test solution witout test material.
- Evidence of undissolved material: no
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Age at study initiation: not older than 24 hours
- Method of breeding: The strain with the parents generation was bred and mantained in vessels containing a lot of Daphnia magna in differents ages. for the study young Dahnia magna were separated in 100 mL of reconstituted water.
- Source: Institute of Toxicology, Merck KGaA, Darmstadt, Germany
- Age of parental stock: differents ages.
- Feeding during test: no
- Aeration during the test: no

ACCLIMATION
- Acclimation conditions: The Daphnia magna were kept in reconstituted water in glass-vessels. The study was located in an air-conditioned room in the Institute of Toxicology. Lighting was controlled by a timer to provide a 16 hours light - 8 hours dark regime.
- Health during acclimation: No mortality was observed during the test .

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
none
Post exposure observation period:
none
Hardness:
250 mg/L expressed as CaCO3
Test temperature:
19 to 20 °C .
pH:
7.86 - 7.97
Dissolved oxygen:
71.2 - 96.7 %
Salinity:
n.a
Conductivity:
n.a.
Nominal and measured concentrations:
- Nominal concentrations: 1, 1.8, 3.2, 5.6 and 10 mg/L
- Measured concentrations: The analytically determined test material concentrations of the freshly prepared samples corresponded to the nominal values of 3.2, 5.6, and 10 mg/L. Following measured concentrations were determined: 3.4, 5.7 and 1 mg/L corresponding to ca. 100 % of the nominal concentrations.
Details on test conditions:
TEST SYSTEM
- Test vessel: 25 mL, all-glass
- Type: closed with a vaulted glass bowl
- Volume of solution: 10 mL
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water according to OECD 202 (1984)
- Culture medium different from test medium: No
- Intervals of water quality measurement: The pH-values and dissolved oxygen concentration (O2) were measured in the control and all test material concentrations at the beginning and at the end of the experimental part. During the experimental part, the temperature was registered continuously in a control vessel with an electronic thermometer containing a maximum and minimum memory display.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light - 8 hours dark regime

EFFECT PARAMETERS MEASURED: Immobility of the animals after 24, 48 hours

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Not performed
Reference substance (positive control):
no
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
5.1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Behavioural abnormalities: not observed
- Observations on body length and weight: not observed
- Other biological observations: not observed
- Mortality of control: no
- Other adverse effects control: not observed
- Immobilisation of control: not observed
- Abnormal responses: not observed
- Any observations that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no
Results with reference substance (positive control):
No test with reference substance was performed.
Reported statistics and error estimates:
The EC50 was calculated as a logit analysis according the procedure of Unkelbach and Wolf (1985) using the PC-program 511.

Table 1: Nominal concentration and immobilization data

Nominal concentration
[mg/l]

Immobilized / exposed Daphnia magna

24 h

48 h

Control

0 / 20

0 / 20

1.0

0 / 20

0 / 20

1.8

0 / 20

0 / 20

3.2

0 / 20

0 / 20

5.6

0 / 20

14 / 20

 

 

 

Validity criteria fulfilled:
yes
Conclusions:
The EC50 (48h) of the test item to Daphnia magna was determined to be 5.1 mg/L (reference 6.1.3 -1) according to OECD 202 under GLP conditions.
Executive summary:

The objective of this study was to determine the acute toxicity of the test material to Daphnia magna according to OECD 202 (1984) under GLP conditions. For this purpose, young Daphnia magna were exposed to aqueous test material concentrations under defined conditions. The study comprised of four vessels per concentration containing five Daphnia magna, i.e. 20 Daphnia per concentration (test medium group). The Daphnia were observed for immobilization up to 48 hours. The study was started with the exposure of Daphnia to test material concentrations of nominal 1, 1.8, 3.2, 5.6 and 10 mg/L in a static open system. The test vessels were covered with a vaulted glass bowl. For analytical determination the samples had to be prepared and diluted. Therefore the analytical controls were carried out with 3.2, 5.6 and 10 mg/L. The measured concentrations were determined as 3.4, 5.7 and 1 mg/L corresponding to ca. 100 % of the nominal concentrations. A blank control without test substance was performed. No test with reference substance as positive control was determined. The pH-values and dissolved oxygen concentration (O2) were measured in the control and all test material concentrations at the beginning and at the end of the experimental part. During the experimental part, the temperature was registered continuously in a control vessel with an electronic thermometer containing a maximum and minimum memory display. The EC50 (48h) of the test item to Daphnia magna was determined to be 5.1 mg/L.

Description of key information

The EC50 (48h) of the test item to Daphnia magna was determined to be 5.1 mg/L (reference 6.1.3-1) according to OECD 202 under GLP conditions.

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
5.1 mg/L

Additional information

The objective of this study was to determine the acute toxicity of the test material to Daphnia magna according to OECD 202 (1984) under GLP conditions. For this purpose, young Daphnia magna were exposed to aqueous test material concentrations under defined conditions. The study comprised of four vessels per concentration containing five Daphnia magna, i.e. 20 Daphnia per concentration (test medium group). The Daphnia were observed for immobilization up to 48 hours. The study was started with the exposure of Daphnia to test material concentrations of nominal 1, 1.8, 3.2, 5.6 and 10 mg/L in a static open system. The test vessels were covered with a vaulted glass bowl. For analytical determination the samples had to be prepared and diluted. Therefore the analytical controls were carried out with 3.2, 5.6 and 10 mg/L. The measured concentrations were determined as 3.4, 5.7 and 1 mg/L corresponding to ca. 100 % of the nominal concentrations. A blank control without test substance was performed. No test with reference substance as positive control was determined. The pH-values and dissolved oxygen concentration (O2) were measured in the control and all test material concentrations at the beginning and at the end of the experimental part. During the experimental part, the temperature was registered continuously in a control vessel with an electronic thermometer containing a maximum and minimum memory display. The EC50 (48h) of the test item to Daphnia magna was determined to be 5.1 mg/L.