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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Juny 15, 2016 - September 2, 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Specific details on test material used for the study:
Designation: Art. 201386
Synonym: B-2O-O5
Batch: EF13004186
Purity (HPLC): 100.0%
Appearance: White, solid
Water solubility: <0.00003 g/L (OECD 105)
Stability under test conditions: Not specified
Date of expiry: April 30, 2018
Storage: Tightly closed, dark at room temperature (15 to 25°C)

Preparation of the Test Item:
A stock preparation with a test item concentration of 100 mg/L was freshly prepared. For that purpose, the test item was weighed in a calibrated flask and vehicle was added. The preparation was treated in an ultrasonic device for 30 minutes. Subsequently, the preparation was stirred with a magnetic stirrer for further 23 hours. Afterwards, the formulation was passed through a membrane filter with a pore size of 0.2 µm. The filtrate was used for the study.

Sampling and analysis

Analytical monitoring:
no
Remarks:
The test item concentration in the reconstituted water was not quantified at the start and the end of this study. Because of the low water solubility (<0.00003 g/L), the compound cannot be detected with standard analytical methods.

Test solutions

Vehicle:
yes
Details on test solutions:
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: 7.89
- Total hardness: 220 mg CaCO3/L

Preparation of the Test Item:
A stock preparation with a test item concentration of 100 mg/L was freshly prepared. For that purpose, the test item was weighed in a calibrated flask and vehicle was added. The preparation was treated in an ultrasonic device for 30 minutes. Subsequently, the preparation was stirred with a magnetic stirrer for further 23 hours. Afterwards, the formulation was passed through a membrane filter with a pore size of 0.2 µm. The filtrate was used for the study.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Species: Daphnia magna Straus
Origin: Daphnia magna Straus was originally obtained from IBACON GmbH (Roßdorf, Germany).
Culture conditions: The clone is bred in the laboratories of Merck KGaA (room 2068).
Parental daphnids are used for reproduction until they are about 6 weeks old. Thereafter, they are replaced by neonates.
Daphnids are kept individually in 100 mL glass vessels containing approximately 60 mL reconstituted water (ELENDT M4 medium) at a water temperature of 20 ± 2°C and a 16 hour light and 8 hour dark regime to ensure similar conditions as in the experiment. Offspring are removed from the vessels at least twice per week.
Feeding: The parental daphnids are fed ad libitum with unicellular green algae Desmodesmus subspicatus three times per week.
Age: Offspring less than 24 hours old were used for the study.
Acclimation period: same as test

Feeding during test: None

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h

Test conditions

Hardness:
220 mg CaCO3/L
Test temperature:
19.2 – 19.9°C
pH:
7.89 - 7.91
Dissolved oxygen:
7.72 - 8.35
Conductivity:
802 µS/cm
Nominal and measured concentrations:
Nominal 100 mg/L
Details on test conditions:
EXPOSURE:
The study was performed in an air-conditioned room. For the study 20 mL glass test tubes containing at least 20 mL either reconstituted water (control group) or test medium (test item group) were used. Each test vessel contained five daphnids resulting in 4 mL medium per daphnid. 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: 7.89
- Total hardness: 220 mg CaCO3/L
- O2-Concentration: 8.35 mg/L
- Conductivity: 802 µS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: 48 h


OTHER TEST CONDITIONS
- Photoperiod: 16h light / 8h dark
- Light intensity: The mean light intensities were 614 Lux prior to and at the end of the exposure period, respectively.

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.

Results and discussion

Effect concentrationsopen allclose all
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 > 3.0E-5 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 > 3.0E-5 g/L
Details on results:
Under the conditions of the present study, an aqueous solution of nominal 100 mg/L of Art. 201386 (B-2O-O5) revealed no aquatic toxicity in the test system.

The 48h EC50 exceeded the water solubility of 0.00003 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
Results with reference substance (positive control):
Potassium dichromate:
24h EC50 1.00 (0.85 - 1.23) mg/L
48h EC50 0.66 (could not be determined) mg/L

The test item Art. 104864 (Potassium dichromate) showed a 24h EC50 value of 1.0 mg/L which is within the range of the published data of 0.6 to 1.7 mg/L (Council Regulation (EC) No. 440/2008) and 0.6 to 2.1 mg/L (OECD Guideline No. 202).

Any other information on results incl. tables

Objective

The objective of this study was to determine the effect of the test item Art. 201386 (B-2O-O5) on the immobilization of Daphnia magna.

Study Design

Juvenile daphnids were exposed to a nominal test item concentration of 100 mg/L (limit test) in an open static system. 20 daphnids, divided into four replicates, each with five animals were used in the test item group and the control group. The daphnids were observed for immobilization after 24 and 48 hours of exposure.

Results

The test item concentration in the reconstituted water was not quantified at the start and the end of this study. Because of the low water solubility (<0.00003 g/L), the compound cannot be detected with standard analytical methods and the development of an analytical method with a sufficiently low detection and quantification limit is complex.

Due to absence of any adverse effects at the saturation concentration, the study was performed without analytical verification.

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

For the test item Art. 201386 (B-2O-O5) the following EC50 values were determined:

EC50 (24h) >0.00003 g/L (nominal >100 mg/L)

EC50 (48h) >0.00003 g/L (nominal >100 mg/L)

Conclusion

Under the conditions of the present study, an aqueous solution of nominal 100 mg/L of Art. 201386 (B-2O-O5) revealed no aquatic toxicity in the test system.


The 48h EC50was >0.00003 g/L (nominal >100 mg/L) and, thus, could not be determined in this study.


Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of the present study, an aqueous solution of nominal 100 mg/L of Art. 201386 (B-2O-O5) revealed no aquatic toxicity in the test system.

The 48h EC50 was >0.00003 g/L (nominal >100 mg/L) and, thus, could not be determined in this study.
Executive summary:

This study was performed according to GLP and the methods applied are fully compliant with OECD TG 202. Under the conditions of the present study, an aqueous solution of nominal 100 mg/L of Art. 201386 (B-2O-O5) revealed no aquatic toxicity in the test system. The 48h EC50 exceeded the water solubility of 0.00003 g/L (nominal >100 mg/L) and, thus, could not be determined in this study.