Cuprate(4-), [[3,3',3'',3'''-[(29H,31H-phthalocyanine-1,8,15,22-tetrayl-.kappa.N29,.kappa.N30,.kappa.N31,.kappa.N32)tetrakis(sulfonyl)]tetrakis[1-propanesulfonato]](6-)]-, sodium (1:4), (SP-4-1)-

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:

2013-12-18 to 2014-01-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study meets the criteria for classification as reliable without restriction according to Klimisch et al (1997).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

The study plan states Daphnia used in testing should be <24h old. Some of those used in the test may be 24h and 9min old. This was not considered to impact the validity of the test. Observations in the definitive test were consistent with rangefinder.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

tetrasodium 1,8,15,22-tetra-(3-sulfoxypropylsulfonyl)copper phthalocyanine

EC Number:

696-145-8

Cas Number:

944730-39-6

Molecular formula:

C44H36CuN8O20S8 .4Na

IUPAC Name:

tetrasodium 1,8,15,22-tetra-(3-sulfoxypropylsulfonyl)copper phthalocyanine

Test material form:

other: solid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 120 mg/l
- Sample storage conditions before analysis: samples were stored frozen prior to analysis

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Reconstituted Water - ISO medium

Based on the results of the range-finding test a "Limit test" was conducted at a concentration of 120 mg/L to confirm that at the highest test concentration no immobilization or adverse reactions to exposure were observed.

An amount of test item (120 mg) was dissolved in test water and the volume adjusted to 1 liter to give the 120 mg/L test concentration.

The prepared concentration was inverted several times to ensure adequate mixing and homogeneity.

The concentration and stability of the test item in the test preparation was verified by chemical analysis at 0 and 48 hours

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

The test was carried out using 1st instar Daphnia magna derived from in-house laboratory cultures.

Adult Daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium in a temperature controlled room at approximately 20 ºC. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were approximately 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

The study plan states that the Daphnia to be used in testing should be less than 24 hours old. However, some of those used in the definitive test may have been greater than 24 hours old as they were previously isolated 24 hours and 9 minutes prior to the start of the test. This was considered not to impact the validity of the test as the observations in the definitive test were consistent with those from the range-finding test.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Test temperature:

20ºC

pH:

7.6-7.9

Dissolved oxygen:

8.8-9.1 mg O2/L

Nominal and measured concentrations:

Analysis of the test preparations at 0 and 48 hours showed measured test concentrations to be near nominal so it was considered justifiable to estimate the EC50 values in terms of the nominal test concentrations only.

Details on test conditions:

250 mL glass jars containing approximately 200 mL of test preparation were used. At the start of the test 5 daphnids were placed in each test and control vessel at random, in the test preparations. Four replicate test and control vessels were prepared. The test vessels were then covered to reduce evaporation and maintained in a temperature controlled room at 20 ºC with a photoperiod of 16 hours light (532 to 546 lux) and 8 hours darkness with 20 minute dawn and dusk transition periods. The daphnids were not individually identified, received no food during exposure and the test vessels were not aerated.

The control group was maintained under identical conditions but not exposed to the test item.

The test preparations were not renewed during the exposure period.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: 0% immobilization

Results with reference substance (positive control):

A positive control used potassium dichromate as the reference item at concentrations of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L.

Exposure conditions for the positive control were similar to those in the definitive test.

Analysis of the immobilization data by the trimmed Spearman-Karber method (Hamilton et al 1977 ) at 24 and 48 hours based on the nominal test concentrations gave the following results:

Time Point
(Hours) EC50(mg/L) 95% Confidence Limits (mg/L) (NOEC) (mg/L) (LOEC) (mg/L)
24 1.0 0.91 - 1.2 0.56 1.0
48 0.71 0.65 - 0.76 0.32 0.56

The No Observed Effect Concentration is based upon zero immobilization at this concentration.

The results from the positive control with potassium dichromate were within the normal range for this reference item.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Exposure of Daphnia magna to the test item gave EC50 values of >120 mg/L. The NOEC was 120 mg/L. Therefore it is concluded that the test material did not have a significant toxic effect on Daphnia magna in this study.

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item toDaphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphniasp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.

Results & Conclusions

Exposure of Daphnia magna to the test item gave EC50 values of >120 mg/L. The NOEC was 120 mg/L. Therefore it is concluded that the test material did not have a significant toxic effect on Daphnia magna in this study.