[No public or meaningful name is available]

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:

From 2018-04-10 to 2018-04-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Bis and tris and tetra{N-[(2-ethylanilino) or 2,4(or 2,5 or 2,6)-dimethylanilino]-N’-[(2-ethylanilino) or 2,4(or 2,5 or 2,6)-dimethylanilino] methaniminium} [phthalocyanine(bis and tris and tetra)sulfonato-κ4N29,N30,N31,N32]cuprate(II)

Cas Number:

265115-84-2

Molecular formula:

C32H16-τN8Cu(SO3C17H22N3)τ

IUPAC Name:

Bis and tris and tetra{N-[(2-ethylanilino) or 2,4(or 2,5 or 2,6)-dimethylanilino]-N’-[(2-ethylanilino) or 2,4(or 2,5 or 2,6)-dimethylanilino] methaniminium} [phthalocyanine(bis and tris and tetra)sulfonato-κ4N29,N30,N31,N32]cuprate(II)

Test material form:

solid: particulate/powder

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
* 0 mg/L of test item (control)
* 0.0500 mg/L of test item
* 0.158 mg/L of test item
* 0.500 mg/L of test item
* 1.58 mg/L of test item
* 5.00 mg/L of test item
* 15.8 mg/L of test item

- Sampling method: Sampling done in freshly prepared media at the start of exposure (0 hours) and in old media at the end of exposure (48 hours) without daphnids.
The control and the lower concentrated samples (0.0500 mg/L and 0.158 mg/L) were analyzed enriched factor 10. The samples 0.500 mg/L and 1.58 mg/L were analyzed undiluted. The higher concentrated samples (5.00 mg/L and 15.8 mg/L) were diluted with dilution medium prior to analysis.

- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until the start of the analysis, if necessary. Prepared samples were stored in the autosampler at room temperature until analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution with a nominal test item concentration of 15.8 mg/Lwas freshly prepared with dilution water one day prior to the start of the exposure (at day -1). This solution was stirred with a magnetic stirrer at approximately 1100 rpm for 24 hours at room temperature.
The four other concentrations: 0.0500 - 0.158 - 0.500 - 1.58 - 5.00 mg/L of test item were prepared by appropriate dilution of the stock solution.

- Controls: dilution water without test item incubated under the same conditions as the test groups.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnid
- Strain/clone: STRAUSS
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany
- Age of parental stock (mean and range, SD): less than 24 hours (at test initiation)
- Feeding during test: no
- Acclimation period: No acclimation period as the daphnids were cultured under the same conditions as the test replicates.

Study design

Test type:

static

Water media type:

other: medium Elendt M4 according to OECD 202, Annex 3 (2004)

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

285 mg CaCo3/L of the Dilution Water at the Start of the Exposure (0 hours)

Test temperature:

18 - 22°C, constant within +/- 1°C

pH:

7.73-7.96 (fresh media at start of the exposure)
7.76-7.65 (in old Media at the End of the Exposure (48 hours))
8.10 Dilution Water at the Start of the Exposure

Dissolved oxygen:

9.30 mg/L - 10.3 (fresh media at start of the exposure)

Nominal and measured concentrations:

Nominal: 0 mg/L; 0.050 mg/L; 0.158 mg/L; 0.500 mg/L; 1.580 mg/L; 5.00 mg/L; 15.8 mg/L
Measured (fresh media 0hr/old media 48h): < LOQ/< LOQ; 0.0441/0.0497 mg/L; 0.134/0.131 mg/L; 0.529/0.516 mg/L; 1.56/1.58 mg/L; 5.08/5.39 mg/L; 15.7/15.6 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity
- Type : closed with watch glasses
- Material, size, headspace, fill volume: filled at 20 mL.
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): no renewal
- No. of organisms per vessel: 5 daphnids
- No. of vessels per concentration (replicates): 4 replicates
- No. of vessels per control (replicates): 4 replicates

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light, 8 hours darkness
- Light intensity: Diffuse light, maximum of 1500 lx

VEHICLE CONTROL PERFORMED: no
RANGE-FINDING STUDY
- Test concentrations:
* 0 mg/l of test item (control)
* 0.0500 mg/l of test item
* 0.5000 mg/l of test item
* 5.00 mg/l of test item
- Results used to determine the conditions for the definitive study: Immobilization rates were used to determine the conditions for the definitive study

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CI 6.47-15.6 mg/l

Effect concentrations 2

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 15.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

ca. 4.48 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CI 0.846- 15.0 mg/l

Effect concentrations 4

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

ca. 1.58 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 5

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 15.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 6

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 15.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: none observed.
- Mortality of control: No mortality observed

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- ECx: EC50 = 2.00 mg/l
- 95% confidence limits: 1.00 - 4.00 mg/L

Reported statistics and error estimates:

EC10 and EC 50 are determined via the following the Sigmoidal dose-response (variable slope) following the equation:
Y=Bottom + (Top-Bottom)/(1+10^((LogEC50-X)*HillSlope))

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentration of the test item, the 48-hour EC50 for Daphnia magna was 13.0 mg/L. No effects on Daphnia magna STRAUS were observed.

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS), the effects of the test item were determined at the test facility according to OECD 202 (2004) from 2018 -04 -10 to 2018-04-16, with the definitive exposure phase from 2018-04-11 to 2018-04-13.

The study was conducted under static conditions over a period of 48 hours with the following nominal test item concentrations: 0.0500 - 0.158 - 0.500 - 1.58 - 5.00 - 15.8 mg/L of the test item.

The test item concentration was blue coloured in a concentration-related manner and visually clear throughout the exposure.

Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each test item concentration and the control.

The concentrations of the test item were analytically verified via HPLC-DAD in fresh media at the start (0 hours) and in old media at the end of the exposure (48 hours) in each test item concentration and in the control.

The measured concentrations of the test item at test start were between 85% and 106% of the nominal test item concentration. At 48 hours, the measured concentrations were between 83% and 108% of the nominal test item concentration.

The measured test item concentrations remained stable within ± 20% of the nominal concentration of the test item over the test period. The effect concentrations are based on the nominal concentrations of the test item.

The validity criteria of the test guideline were fulfilled.

Based on the nominal concentration of the test item, the 48 -hour EC50 for Daphnia magna was 13.0 mg/L.