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:
04 - 06 Jun 2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guidelineopen allclose all
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)
Remarks:
Niedersächsisches Umweltministerium, Hannover, Germany

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: control and 100 mg/L (Limit concentration which can be achived by suitable preparation methods)

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution was prepared one day prior to application at a concentration of 100 mg/L. The stock solution was shaken with 20 rpm for 24 h. Undissolved particles were removed by centrifugation (20 min at 3000 rpm).
- Eluate: no
- Differential loading: no
- Controls: yes, test medium control

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Source: Institut für Wasser-, Boden- und Lufthygiene des Bundesgesundheitsamt Berlin, Germany
- Feeding during test: none
- Culture: Daphnids were cultured in 2-3 L glass vessels with approx. 1.8 L culture medium (M4-medium) at 21 °C (20-25 °C); 16 h illumination (20 µmol*m-2*s-1); approx. 1200 lux. Daphnids were fed ad libitum with a mix of Scenedesmus subspicatus and Chlorella vulgaris with a cell density of >10E+06 cells/mL.
- Age at study inititation: 2-24 h; obtained by removing the mother animals twice within 22 h

ACCLIMATION
- Acclimation period: 2 h in dilution water

Study design

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

Test conditions

Hardness:
252 mg CaCO3/L (dilution water)
Test temperature:
21.2 - 21.9 °C
pH:
7.34 - 7.50
Dissolved oxygen:
7.80 - 8.06 mg/L
Conductivity:
668 µS/cm (dilution water)
Nominal and measured concentrations:
Nominal: control, saturated solution
measured DOC: please refer to "Any other information on results incl. tables"
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers (5 cm i.d. x 8 cm), volume: 50 mL, headspace: 30 mL, fill volume: 20 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: according to EEC 92/69 L383A C.2
- Culture medium different from test medium: same as test

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light / 8 h dark
- Light intensity: approx. 1200 lux

EFFECT PARAMETERS MEASURED
- Immobilisation after 24 and 48 h

RANGE-FINDING STUDY
- Test concentrations:
- Results used to determine the conditions for the definitive study:
Reference substance (positive control):
yes

Results and discussion

Effect concentrations
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Reported as "saturated solution"
Details on results:
- Mortality of control: 0%
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Dose-response test: yes, 0.58 - 5.8 mg/L
- ECx: EC50 (24 h): 1.5 mg/L (95% CI 1.3 - 1.7 mg/L)
Reported statistics and error estimates:
Effect concentrations were derived directly from the test results.

Any other information on results incl. tables

All validity criteria were met:

1.) pH did not vary by more than 1 unit

2.) No immobilisation was recorded in the control.

3.) EC50 value of the reference substance was between 1.0 - 2.5 mg/L.

4.) Dissolved O2 concentration was > 5.21 mg/L (60% of air saturation level)

Table 1: Results from DOC analysis

 

DOC [mg/L]

0 h (new without daphnids)

48 h (old without daphnids)

48 h (old media with daphnids)

Overall mean

Corrected vs. control

Saturated solution

43.9

43.37

51.65

46.31

41.16

Control

3.22

3.06

9.17

5.15

--

Applicant's summary and conclusion

Validity criteria fulfilled:
yes