2,20-dichloro-13,31-diethyl-4,22-dioxa-13,18,31,36-tetraazanonacyclo[19.15.0.03,19.05,17.06,14.07,12.023,25.024,32.025,30]hexatriaconta-1(36),2,5,7,9,11,14,16,18,20,23,25,27,29,32,34-hexadecaene

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study according to OECD Guideline 202 and Directive 92/69/EC Method C.2

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

GLP according to German "Chemikaliengesetz", EC and OECD Guideline principles

Analytical monitoring:

no

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 was prepared with dilution water one day day prior to application.
Preparation of the saturated solution: A dispersion (100 mg/L test item was weighed out) was prepared with dilution water, shaken with 20 rpm for 24 h (rotating shaker 3040, GFL) and membrane filtrated with 0.45µm (RC Schleicher & Schuell).

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 μE⋅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
- Type and amount of food: a mix of Desmodesmus subspicatus and Chlorella vulgaris with an algae cell density of > 100 000 cells/mL
- Feeding frequency: at least 5 x weekly ad libitum

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

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

none

Hardness:

249 mg CaCO3/L in dilution water prior to test start

Test temperature:

20 ± 1 °C constant

pH:

7.67 (saturated solution) and 7.85 (controls) at the start
7.66-7.72 (saturated solution, 4 replicates) and 7.60-7.65 (controls, 4 replicates) at the end (48 h)

Dissolved oxygen:

8.47 (saturated solution) and 8.66 (controls) mg/L at the start
7.89-7.95 (saturated solution, 4 replicates) and 7.87-8.06 (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 test item was clearly dissolved in the tested limit concentration throughout the whole exposure period. 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 μE⋅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

Reference substance (positive control):

yes

Remarks:

potassium dichromate p.a. (Merck)

Duration:

48 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:

EC0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

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:
- EC10 (immobilisation): 0.97 mg/L
- EC50 (immobilisation): 2.07 mg/L (95% confidence interval: 1.68-2.56 mg/L)
- Other:
EC50 values determined by sigmoidal dose-response regression.
EC100 (immobilisation): 5.80 mg/L (determined directly from the test results)

Reported statistics and error estimates:

EC10- and EC50-values were only calculated for the reference item via probit analysis according to WEBER (1986). Calculation of the confidence intervals for EC50 were carried out using standard procedures according to BREITIG and TÜMPLING.W (1982).
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: SigmaPlot rel. 2000 (2000), SPSS CORPORATION

Reference:
Breitig G. & Tümpling W. (1982): Methoden der Wasseruntersuchungen Bd. II, VEB Gustav Fischer Verlag, Jena

Validity criteria fulfilled:

yes

Conclusions:

At the saturated solution (nominally 100 mg/L) 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 (nominally 100 mg/L) of Pigment Violet 23, was determined according to OECD 202 (1999, Draft) and Directive 92/69/EC Method C.2 (1992). The limit test was conducted under static conditions over 48 h from 2004 -11 -09 to 2004-11-11 at DR.U.NOACK-LABORATORIEN, Käthe-Paulus-Straße 1, D-31157 Sarstedt. 20 test organisms were exposed to the saturated solution and the control. 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 2.07 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 no biologically significant effect was determined.

Description of key information

In the acute immobilisation test with Daphnia magna (STRAUS) the effect of the saturated solution (nominally 100 mg/L) of the test item (Pigment Violet 23), was determined according to OECD 202. At the saturated solution of the test item (Pigment Violet 23) no biologically significant effect was determined.

Additional information

In the acute immobilisation test with Daphnia magna (STRAUS) the effect of the saturated solution (nominally 100 mg/L) of the test item was determined according to OECD 202 (1999, Draft) and Directive 92/69/EC Method C.2 (1992). The limit test was conducted under static conditions over 48 h.

At the saturated solution of the test item no biologically significant effect was determined.