Registration Dossier

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:
02 - 18 Aug 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 guideline
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
13 Apr 2004
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Landesanstalt für Umwelt, Messungen und Naturschutz Baden-Württemberg, Karlsruhe, Germany

Test material

Reference
Name:
Unnamed
Type:
Constituent

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: control, and 100 mg/L (nominal)
- Sampling method: Analytical samples were taken at test start and after 24 h from fresh and aged solutions and after 48 h from aged solutions in one separate replicate per test item concentration without test organisms. For each sampling also a retain sample was taken.
- Sample storage conditions before analysis: All samples were stored deep frozen until they were transferred to the analytical laboratory.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The necessary amount of test item for preparing the stock solution S1 was weighed on a weighing scoop and transferred to a volumetric flask. Test medium was added up to the bench mark and the solution was homogenised by shaking and treated with two times 5 minutes of ultrasonication. The preparation procedure was repeated after 24 h. 50 mL of the prepared solutions were transferred to each test vessel.
- Differential loading: No. Lower test solutions were prepared by dilution of the appropriate solutions with test medium.
- Controls: Yes, untreated test medium.
- Evidence of undissolved material: After homogenisation, the stock solution was clear and transparent and showed foaming. The substance solved hardly from the vessel and small pellets of substance were built during the treatment with ultrasonication.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain/clone: clone V
- Source: In-house culture, originally purchased from Federal Environment Agency, Berlin, Germany
- Age of Daphnia at test start: Freshly hatched Daphnia, less than 24 h old Daphnia were used for the test.
- Feeding during test: No feeding during test.


BREEDING CONDITIONS
- Breeding: D. magna was bred as single culture (1 daphnid per 100 mL) in Elendt M4 medium. The medium was changed three times per week. A pipette was used to separate the young daphnids from the adults.
- pH of the aerated water: 6.0 - 9-0
- Dissolved oxygen > 60% saturation
- Total hardness: 140 - 250 mg/L CaCO3 (7.8 - 14 °dH)
- Temperature: 20 ± 2 °C (climatic chamber)
- Photoperiod: 16 h light/8 h darkness.
- Food type: Single cell green algae (Desmodesmus subspicatus)
- Frequency: At least three times a week.

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
232 mg/L CaCO3, corresponding to 13 °dH (test medium Elendt M4)
Test temperature:
19.4 - 21.5 °C
pH:
7.67 - 8.12
Dissolved oxygen:
≥ 8.3 mg/L
Nominal and measured concentrations:
control, 6.25, 12.5, 25, 50, and 100 mg/L (nominal)
control, NA, NA, NA, NA, and 96.2 - 99.4% of nominal (measured)
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 mL glass beakers covered with glass plates
- Fill volume: 50 mL
- Aeration: None.
- Renewal rate of test solution: After 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Culture medium different from test medium: No, Elendt M4 medium was used as test medium.
- Intervals of water quality measurement: The test temperature and the pH-value as well as oxygen concentration of the test solutions were measured at all concentrations at t = 0 h and at t = 24 h in fresh test solutions and after t = 24 h and t = 48 h in aged test solutions in one separate replicate per test item concentration without test organisms.

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

EFFECT PARAMETERS MEASURED:
Immobilisation: After 24 h and 48 h

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: Yes (non-GLP)
Reference substance (positive control):
yes
Remarks:
potassium dichromate

Results and discussion

Effect concentrations
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Mortality of control: 0%
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Relevant effect levels: EC50 (24 h): 1.0 - 2.0 mg/L (nominal, potassium dichromate)
Reported statistics and error estimates:
Since no effects were observed, no statistical analysis was performed.

Any other information on results incl. tables

ANALYTICAL RESULTS

TOC analysis of the test solutions showed that the measured fresh TOC content for the concentrations ranged from 96.2 - 98 .8% of nominal. In the aged sample the measured TOC content was 99.4% of nominal. Since the measured TOC content of concentrations relevant for the determination of the toxicological endpoints were within 80 - 120% of nominal concentration, toxicological endpoints were evaluated using the nominal concentrations of the test item.

BIOLOGICAL RESULTS

After 48 h no immobilisation was observed up to and including 6.25 mg/L and 25 mg/L and in the test item concentration of 100 mg/L (nominal). 5% of immobilisation was observed at 12.5 and 50 mg/L, which is below the validity criterion of max. 10% immobilisation in the control treatment, as defined by the guideline.

Applicant's summary and conclusion