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:
2008-11-04 to 2008-11-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, guideline study
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
, 2004
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:
signed 2006-06-01

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Naringin dihydrochalcone (DHC)
- Physical state: solid, whitish powder
- Storage condition of test material: room temperature, dark and dry
No further details are given.
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No other test material was used.

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Analytical evaluation of various concentrations of the test item Naringin Dihydrochalcone (DHC), was carried out via HPLC-DAD.
- Concentrations: The samples of the limit concentration were diluted with mobile phase prior to analysis. The nominal test item concentration of 100 mg/L was diluted by the factor 50. The sample volume and the final volume were 0.2 mL and 10 mL, respectively.
- Sample storage conditions before analysis: All samples were stored at 6 +/- 2°C until start of analysis if necessary.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The stock solution (100 mg/L test item was weighed out) was freshly prepared with dilution water. The stock solution was treated with ultrasound for 15 minutes and stirred with approximately 1100 rpm at room temperature.
- Controls: Dilution water without test item tested under the same conditions as the test group.
No further details are given.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Strain: Daphnia magna STRAUS (Clone 5)
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
- Age at study initiation: 2-24 hours old
- Method of breeding: in glass vessels with approximately 1.8L culture medium, at 20 +/- 2°C, in an incubator, 16 hours illumination
- Feeding during test: The daphnids were not fed during the study.

ACCLIMATION
- Acclimation period: at least 2 hours in dilution water
- Type and amount of food: A mix of unicellular green algae (Pseudokirchneriella subcapitata, Desmodesmus subspicatus and Chlorella vulgaris, density of green algae was >10^6 cells/mL) was given.
- Feeding frequency: at least 5 times per week ad libitum
- no mortality was observed

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
not applicable

Test conditions

Hardness:
272 mg CaCO3/L
Test temperature:
19-20°C
pH:
test start, 0 hours: pH 7.19 (test item) and pH 7.69 (control)
test end, 48 hours: pH 7.32 to pH 7.34 (test item) and pH 7.31 to pH 7.32 (control)
see also attached tables
Dissolved oxygen:
test start, 0 hours: 8.53 mg/L (test item) and 8.11 mg/L (control)
test end, 48 hours: 7.89 to 8.05 mg/L (test item) and 8.0 to 8.06 mg/L (control)
see also attached tables
Salinity:
According to Directive 92/69/EC L383A C.2. Annex
KCl: 5.76 mg/L
NaHCO3: 64.8 mg/L
CaCl2 x 2H2O: 294 mg/L
MgSO4 x 7H2O: 123 mg/L
Nominal and measured concentrations:
nominal concentration: 100 mg/L;
measured concentration at test start: 103 mg/L;
measured concentration at test end: 96.0 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass beakers (4 cm ID x 7 cm H), 50 mL capacity; 20 mL volume per vessel
- No. of organisms per vessel: 20 animals, divided into 4 parallel samples, each with 5 animals per limit concentration and control.
- No. of vessels per concentration: duplicates
- No. of vessels per control: duplicates

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dilution water was prepared according to Directive 92/69/EEC Method C.2, Annex 1
- Intervals of water quality measurement: Prior to test start the water quality parameters of the dilution water were measured. At the beginning of the test the water quality parameters of the fresh media were measured and at test end after 48 hours, the water quality parameters of the old media were measured.

OTHER TEST CONDITIONS
- Photoperiod: 16/8 hours light/dark cycle
- Light intensity: max 20 µE x m-2 x s-1

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: limit test, only one concentration is required
- Range finding study
- Test concentrations: 190 and 380 mg/L (calculated water solubility)
- Results used to determine the conditions for the definitive study: 0% immobilisation was observed under static test conditions after 24 or 48 hours with test concentrations of 190 and 380 mg/L.

No further details are given.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (>99.5% pure)

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
LOEC
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
There were no effects, neither in the limit concentration nor in the control group.
Results with reference substance (positive control):
- Results with reference substance valid? yes; The 24-hour EC50-value for potassium dichromate was found to be within the prescribed concentration range of 1.0 - 2.5 mg/L which is in line with the quality criteria stated in AQS P 9/2 (05/1996) for daphnids clone 5. The EC50-value of the reference item is also within the recommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.
- EC50/LC50: 1.5 mg/L (CI 1.3 - 1.73 mg/L)
Reported statistics and error estimates:
The effects of the limit concentration of 100 mg/L were determined directly from the test results.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
see "overall remarks"
Conclusions:
There was no effect on immobilisation of Daphnia magna at the limit concentration of 100 mg/L (nominal) of the test item Naringin dihydrochalcone (DHC).