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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29 May 2006 to 24 July 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study according to OECD Guideline 202 and EU Method C.2

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

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)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
GLP according to German "Chemikaliengesetz", EC and OECD Guideline principles

Test material

Reference
Name:
Unnamed
Type:
Constituent

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances):
A saturated solution was tested in a limit test.
Saturated solution: The maximum dissolved concentration of the test item that can be achieved under the test conditions in the test medium, acc. to OECD Series, No. 23 (2000)6.
The solution (100 mg/L test item was weighed out) was prepared with dilution water one day day prior to application.
The stock solution was stirred with ~ 1100 rpm for 24 h. Undissolved particles were removed by membran filtration (0.45¿m)

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Strain: Daphnia magna STRAUS (Clone 5)
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), Berlin, Germany; Breeder: Dr. U. Noack-Laboratorien, Sarstedt, Germany
- Age at study initiation (mean and range, SD): 2-24 h
- Method of breeding:
Culture: In 2-3 L glass vessels with approximately 1.8 L culture medium, at 20 ± 2 °C, in an incubator, 16 h illumination, illumination strength max. 20 ¿E¿m-2 ¿s-1.
Culture medium: Elendt M4, according to Elendt (1990), modified to a total hardness of 160 to 180 mg CaCO3/l.
- Feeding during test: no feeding during the study

ACCLIMATION
- Acclimation period: at least 2 h in dilution water

Reference:
Elendt, B. P. (1990): Selenium Deficiency in Crustacea, Protoplasma, 154, 25-30

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
none

Test conditions

Hardness:
245 mg CaCO3/L in dilution water prior to test start
Test temperature:
18 - 22 °C, ± 1 °C constant
pH:
7.67 (saturated solution) and 7.65 (controls) at the start
7.55-7.61 (saturated solution, 4 replicates) and 7.62-7.80 (controls, 4 replicates) at the end (48 h)
Dissolved oxygen:
8.27 (saturated solution) and 8.52 (controls) mg/L at the start
8.26-8.33 (saturated solution, 4 replicates) and 8.43-8.55 (controls, 4 replicates) mg/L at the end (48 h)
Nominal and measured concentrations:
A limit test with a saturated solution was carried out.
The saturated solution and the control after 0 h (new media) and 48 h (old media) were analytically verified via DOC analysis according to DIN EN 1484 (DOC of saturated solution: 1.8 mg/L). Separate replicates without test organisms for the test item analysis after 48 h were prepared and incubated under test conditions.
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers (5 cm ID x 8 cm H), 50 mL volume were used.
- 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: dilution water according to EU Method C.2, Annex

OTHER TEST CONDITIONS
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: diffuse light, illumination range max. 20 ¿E¿m-2 ¿s-1.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation at 24 and 48 h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Limit test with saturated solution (see above)
- Justification for using less concentrations than requested by guideline: limit test
- Range finding study
- Test concentrations: dilutions of the saturated solution of 1:1, 1:10 and 1:100
- Results used to determine the conditions for the definitive study: Based on the absence of any immobilisation, the main study was conducted as a limit test with the saturated solution.
Reference substance (positive control):
yes
Remarks:
potassium dichromate p.a.

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
Remarks on result:
other: Due to poor solubility of test item, a saturated solution had been prepared using a nominal concentration of 100 mg/L
Details on results:
No immobilisation after 24 and 48 h with the test substance (saturated solution) or the control.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Mortality:
- EC50 (immobilisation): 1.95 mg/L (95% confidence interval: 1.71-2.16 mg/L)
- Other:
EC50 values determined by sigmoidal dose-response regression.
EC100 (immobilisation): 2.87 mg/L (determined directly from the test results)
Reported statistics and error estimates:
EC10- and EC50-values were only calculated for the reference item by sigmoidal dose-response regression. Calculation of the confidence intervals for EC50 were carried out using standard procedures according to CLOPPER and PEARSON (1934).
All data were computer generated and rounded for presentation from the full derived data. Consequently, if calculated manually based on the given data minor variations may occur from these figures. Calculations were carried out using software: GraphPad Prism4 (2005), GRAPHPAD SOFTWARE, INC.

Reference:
Clopper and Pearson (1934): Biometrika, 26: 404-413 citied in GraphPad Prism Statistics Guide 4.0.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
At the saturated solution of the test item no biologically significant effect was determined.
Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS) the effect of the saturated solution of the test item was determined according to OECD 202 (2004) and Directive 92/69/EC Method C.2 (1992). The limit test was conducted under static conditions over 48 h from 2006-06-13 to 2006-06-15 at DR.U.NOACK-LABORATORIEN, Käthe-Paulus-Straße 1, D-31157 Sarstedt. 20 test organisms were exposed to the saturated solution and the control. Details of the analytical results are given in part 9. A reference test was carried out with potassium dichromate to determine the toxicity of the reference item. The EC50-value of the reference item of 1.95 mg/L after 24 h was within the prescribed concentration range of 1.0 to 2.5 mg/L according to AQS. Water quality parameters pH-value and dissolved oxygen concentration, measured at 0 and 48 h, were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled.

At the saturated solution of the test item (Pigment Red 122) no biologically significant effect was determined.