Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

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:
March 24, 2000 - July 13, 2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reference:
Composition 0
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
April 4, 1984
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
Official Journal No. L 383 A, December 29, 1992
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test material information:
Composition 1
Specific details on test material used for the study:
Batch No.: E99988541
Appearance: white crystals






Analytical monitoring:
yes
Details on sampling:
Samples of the filtrate (concentration of 0.1 g/L nominally) were taken for analyses directly after preparation and at the end of the exposure period. At each time point, two samples of about 20 ml filtrate were filled in siliconized glass bottles. They were stored deeply frozen (at least –18°C) until analyses.
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 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 preparation was given through a nutsch filter (pore size >10 - <16 µm). The filtrate of the test material preparation was used for the study.

Test organisms (species):
Daphnia magna
Details on test organisms:
Species: Daphnia magna Straus
Culture 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.
Temperature and humidity in the experimental room was measured using a thermo-hygrograph.
At the start of the experimental phase 5 Daphnia magna were introduced in each test vessel (25 ml, all-glass) containing 10 ml reconstituted water with or without test material. The vessels were open. The test medium was freshly prepared before the introduction of the young Daphnia magna. The Daphnia magna were not fed, and the test medium was not aerated during the test.
Feeding: The parental daphnids are fed ad libitum with unicellular green algae Desmodesmus subspicatus
Age: Offspring less than 24 hours old were used for the study.
Acclimation period: same as test

Feeding during test: None

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Test temperature:
21°C
pH:
8.0
Dissolved oxygen:
95.5 - 99.4%
Nominal and measured concentrations:
Nominal 100 mg/L (measured: <0.00727 g/L)
Details on test conditions:
EXPOSURE:
The study was performed in an air-conditioned room. For the study 25 mL glass vessels containing at least 10 mL either reconstituted water (control group) or test medium (test item group) were used. Each test vessel contained five daphnids. They were not fed and the media were not aerated during the exposure.

The test was performed as a static test in open vessels.

The duration of exposure was 48 hours. During the exposure period, the mobility of the daphnids was assessed daily, i.e. after 24 and 48 hours.

NO. OF DAPHNIDS:
Control Group: 20 daphnids
100 mg/L: 20 daphnids

CONCENTRATION(S)
In a pre-test no immobilization was observed at a concentration of 100 mg/L under open static conditions. Therefore, the solution of a nominal test item concentration of 100 mg/L was tested in the present study.

For the control, reconstituted water (ELENDT M4 medium) was used.

VEHICLE
Reconstituted water (ELENDT M4 medium) was used as vehicle.

Macro nutrients (mg/L)
CaCI2 •2H20 293.80
MgSO4•7H20 123.30
NaHCO3 64.80
KCI 5.80
Na2SiO3•9H20 10.00
NaNO3 0.27
KH2PO4 0.14
K2HPO4 0.18

Trace elements (mg/L)
B 0.5000
Fe 0.2000
Mn 0.1000
Li, Rb and Sr 0.0500
Mo 0.0250
Br 0.0125
Cu and Zn 0.0063
Co and I 0.0025
Se 0.0010
V 0.0003

Macro nutrients (mg/L)
Na2EDTA • 2H20 2.50

Vitamins (µg/L):
Thiamine 75.00
B12 1.00
Biotin 0.75

- pH: 8.0
- O2-Concentration: 99.4%
- Culture medium different from test medium: no
- Intervals of water quality measurement: 48 h


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

References:

ELENDT, B.-P. Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus.
Protoplasma 154, 25-33, 1990
Reference substance (positive control):
no
Remarks:
No positive control used in this study. The accuracy and reliability of the test method is demonstrated periodically as recommended by guidelines with potassium dichromate.
Key result
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: EC50 > 0.00727 g/L
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: EC50 > 0.00727 g/L
Details on results:
The 48 hour EC50 for the test material to Daphnia magna could not be quantified due to the absence of toxicity at the highest test concentration achievable in water. An aqueous filtrate of 0.1 g/L nominally, corresponding to an analytical concentration <0.00727 g/L, revealed no aquatic toxicity in the test system.


The 48h EC50 exceeded the water solubility of <0.00727 g/L (nominal >100 mg/L) and, thus, could not be determined in this study.

- Behavioural abnormalities: no
- Other biological observations: no
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no

Study Design

The objective of this study was to determine the acute toxicity of an aqueous solution of the test material in a static test to Daphnia magna. For this purpose, juvenile Daphnia magna were exposed to an aqueous filtrate of the test item over 48 hours, under defined conditions. The study comprised four vessels per dose group containing five Daphnia magna, i.e. 20 Daphnia per concentration (test medium group and control group). Daphnia magna were exposed to a filtrate (nominal test material concentration of 0.1 g/L) in an aqueous, open, and static system. The study was performed as limit test.

Results

Samples for analysis were taken from the filtrate directly after preparation and at the end of the exposure period. The analytically determined test material concentrations were below the limit of quantification of about 0.00727 g/L.

No effect on the mobility of the daphnia was observed at a nominal concentration of 100 mg/L.

For the test item the following EC50 values were determined:

EC50 (24h) >0.1 g/L

EC50 (48h) >0.1 g/L

Conclusion

The 48 hour EC50 value for the test material to Daphnia magna could not be quantified due to the absence of toxicity at the highest test concentration achievable in water. An aqueous filtrate of 0.1 g/L nominally, corresponding to an analytical concentration <0.00727 g/L, revealed no aquatic toxicity in the test system.

Validity criteria fulfilled:
yes
Conclusions:
The 48 hour EC50 for the test material to Daphnia magna could not be quantified due to the absence of toxicity at the highest test concentration achievable in water. An aqueous filtrate of 0.1 g/L nominally, corresponding to an analytical concentration <0.00727 g/L, revealed no aquatic toxicity in the test system.
Executive summary:

This study was performed according to GLP and the methods applied are fully compliant with OECD TG 202. The 48 hour EC50 for the test material to Daphnia magna could not be quantified due to the absence of toxicity at the highest test concentration achievable in water. An aqueous filtrate of 0.1 g/L nominally, corresponding to an analytical concentration <0.00727 g/L,  revealed no aquatic toxicity in the test system.

Description of key information

Under the given experimental conditions, the test material showed a nominal 48 h EC50 value greater than 100 mg/L.

Key value for chemical safety assessment

Additional information