Tetrasodium [2-({4-fluoro-6-[(2-{[4-fluoro-6-({5-(hydroxykappaO)-6-[(2-{[2-(hydroxy-kappaO)-5-sulfophenyl] diazenyl-kappaN1}-4,5-dimethoxyphenyl) diazenyl-kappaN2]-7-sulfo-2-naphthyl} amino)-1,3,5-triazin-2-yl] amino} propyl) amino]-1,3,5-triazin-2-yl} amino) benzene-1,4- disulfonato(6-)] cuprate(4-)

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

Study period:

Experiment start date - 12 June 2006;
Experiment completion date - 12 July 2006;
Study completion date - 3 August 2006;

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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)

Specific details on test material used for the study:

Identity: FAT 40825/A
Batch number: CHU 297 / BOP 04/05
Purity: Organic part (Na-salt): approx. 83.7 %; All coloured components: approx. 80.54 %; Main component: approx. 59.1 %.
Appearance: Solid, black powder
Storage conditions: At room temperature at about 20 °C
Expiration date: December 31, 2010

Analytical monitoring:

yes

Details on sampling:

For the determination of the actual FAT 40825/A concentration, the following samples were taken:
Just before the start of the test:
- duplicate samples from the test medium (without daphnids)
- duplicate samples from the control (without daphnids)
After 48 hours:
- duplicate samples from the test medium (stability samples)
- duplicate samples from the control
For the 48-hour stability samples, the contents of the respective replicates were combined prior to sampling.
All samples were deep-frozen (at about -20 °C) immediately after sampling. Based on pre-experiments for investigation of the storage stability (without GLP), the test item was found to be stable in the test water under these storage conditions. The concentration of FAT 40825/A was analysed in the duplicate test medium samples of the single test concentration for both sampling times (0 and 48 hours). From the control only one of the duplicate samples was analysed from each of the sampling times.

Vehicle:

no

Details on test solutions:

The test medium was prepared by dissolving 30.1 mg of test item completely in 300 mL of test water using ultrasonic treatment for 10 minutes and intense stirring for 10 minutes at room temperature. The test medium was prepared just before introduction of the daphnids (i.e., start of the test).

Test organisms (species):

Daphnia magna

Details on test organisms:

The study was performed with young daphnids of a clone of the species Daphnia magna Straus. A clone of this species (defined by the supplier as clone 5) was originally supplied by the University of Sheffield/UK in 1992. Since that time, the clone has been bred in the laboratories of RCC in reconstituted water of the quality identical to the water quality used in the tests (in respect to pH, main ions, and total hardness) and under temperature and light conditions identical to those of the tests. At the start of the test, the organisms used in the test were 6 - 24 hours old and were not first brood progeny.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

The water temperature was 20 °C

pH:

The pH values of the test medium were between 7.7 and 7.8.

Dissolved oxygen:

At the beginning and the end of the test period, the dissolved oxygen concentration in the test medium and the control was at least 9.2 mg/L.

Salinity:

no data

Nominal and measured concentrations:

nominal: 100 mg/L
measured: between 99 and 101 % of the nominal value

Details on test conditions:

Experimental Conditions
The test was performed in 100 mL glass beakers filled with 50 mL of test medium. The beakers were covered with glass plates to reduce the loss of water and to avoid the entry of dust into the solutions. The test vessels were labelled with the RCC study number and all necessary additional information to ensure unmistakable identification. At the test concentration and for the control, 20 daphnids were used divided into four replicates of five daphnids each. The daphnids were randomly distributed to the test vessels at initiation of the test. The loading rate was less than one daphnia per 5 mL of test solution. A static test without test medium renewal was chosen since in a pre-experiment (without GLP) the test item concentration in the test medium was constant during the test period of 48 hours. The test was performed under the following conditions:
Water temperature: 20 °C during the test period. The test was performed in a temperature-controlled room (room temperature continuously monitored).
Light conditions: A 16-hour light to 8-hour dark photoperiod (with a 30 minute transition period). Light intensity during the light period was between approximately 470 and 640 Lux.
Feeding: None

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 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:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

In the control and in the test concentration of 100 mg/L, no immobilized test organisms were observed during the test period of 48 hours. No remarkable observations were made concerning the appearance of the test medium. The test medium was a clear solution coloured by the test item throughout the whole test duration.

Results with reference substance (positive control):

The latest result of the positive control test in September 2005 (48-hour EC50: 0.81 mg/L, RCC Study No. A28102) showed that the toxic performance was valid and within the historical range of the RCC laboratory (from 1996 to 2005: 48-hour EC50: 0.53-1.1 mg/L).

Reported statistics and error estimates:

The NOEC and ECO were determined directly from the raw data. The EC50 and the EC 100 could not be determined due to the absence of a toxic effect of the test item in this test.

Validity criteria fulfilled:

yes

Conclusions:

The 48-hour EC50 was higher than 100 mg/L.

Executive summary:

The acute toxicity of the test item FAT 40825/A to Daphnia magna was determined in a 48-hour static test according to the EU Commission Directive 92/69/EEC, Part C.2 (1992), and the OECD Guideline for Testing of Chemicals, No. 202 (2004) in a GLP-certified laboratory. A limit test was performed in accordance with the test guidelines to demonstrate that the test item has no toxic effect on the test organisms up to and including a nominal concentration of 100 mg/L. Thus, the only concentration tested was nominal 100 mg/L and a control. At the start and the end of the test, the analytically determined concentration of the test item in the analyzed test medium was between 99 and 101 % of the nominal value. Thus, the test item FAT 40825/A was stable during the test period of 48 hours under the conditions of the test, and the reported biological results are based on the nominal concentration of the test item. In the control and in the test concentration of 100 mg/L, no immobilized test organisms were observed during the test period of 48 hours. Therefore, the 48-hour NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours) and the 48-hour EC0 of FAT 40825/A to Daphnia magna were determined to be at least 100 mg/L. The 48-hour NOEC and the 48-hour EC0 is even higher but concentrations above 100 mg/L were not tested, in accordance with the test guidelines. The 48-hour EC50 and the 48-hour EC100 were clearly higher than 100 mg/L. These values could not be quantified due to the absence of toxicity of FAT 40825/A at the test concentration of 100 mg/L.

Description of key information

The 48-hour NOEC and the 48-hour EC0 of FAT 40825/A to Daphnia magna were determined to be at least 100 mg/L. The 48-hour EC50 and the 48-hour EC100 were clearly higher than 100 mg/L due to the absence of toxicity of FAT 40825/A at the test concentration of 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

LC50

Effect concentration:

> 100 mg/L

Additional information

The acute toxicity of the test item FAT 40825/A to Daphnia magna was determined in a 48-hour static test according to the OECD Guideline for Testing of Chemicals, No. 202 (2004). A limit test was performed in accordance with the test guidelines to demonstrate that the test item has no toxic effect on the test organisms up to and including a nominal concentration of 100 mg/L. Thus, the only concentration tested was nominal 100 mg/L and a control. At the start and the end of the test, the analytically determined concentration of the test item in the analyzed test medium was between 99 and 101 % of the nominal value. Thus, the test item FAT 40825/A was stable during the test period of 48 hours under the conditions of the test, and the reported biological results are based on the nominal concentration of the test item. In the control and in the test concentration of 100 mg/L, no immobilized test organisms were observed during the test period of 48 hours. Therefore, the 48-hour NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours) and the 48-hour EC0 of FAT 40825/A to Daphnia magna were determined to be at least 100 mg/L. The 48-hour NOEC and the 48-hour EC0 is even be higher but concentrations above 100 mg/L were not tested, in accordance with the test guidelines. The 48-hour EC50 and the 48-hour EC100 were clearly higher than 100 mg/L. These values could not be quantified due to the absence of toxicity of FAT 40825/A at the test concentration of 100 mg/L.