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 fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experiment start date - 15 August 2006;
Experiment completion date - 06 September 2006;
Study completion date - 29 September 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

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 analysis of the actual test item concentration the following samples were taken just before test start:
- duplicate samples from the test medium
- duplicate samples from the control
after 48 and 96 hours:
- duplicate samples from the test medium (stability samples)
- duplicate samples from the control
All samples were taken from the approximate center of the aquaria without mixing the test medium, and 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 is stable in the test water under the storage conditions. The concentration of the test item was analyzed in the duplicate test medium samples from all sampling times. From the control samples only one of the duplicate samples was analyzed from the start and the end of the test.

Vehicle:

no

Details on test solutions:

At the start of the test, the single test concentration of nominal 100 mg/L was freshly prepared by completely dissolving 500.9 mg of the test item in 5000 mL of test water using stirring for 15 minutes at room temperature. The test medium was freshly prepared just before introduction of the fish (= start of the test).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The study was performed with zebra fish (Brachydanio reno). The test fish were raised in the RCC laboratories without any medication. Prior to test start, they were acclimated for one week to the test water and temperature. During holding and acclimatization until one day before the start of the test the fish were fed with a commercial fish diet (Tetra Min® Hauptfutter, supplied by TETRA-Werke, D-49304 Melle, Germany). During the last two weeks prior to the test no fish died in the test fish batch and all fish were healthy. From the acclimated test fish batch, 10 fish were measured at the start of the test: The mean body length of the fish was 3.2 ± 0.14 cm (Mean ± SD), the mean body wet weight was 0.33 ± 0.04 g (Mean ± SD).

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

no data

Test temperature:

The water temperature was 22 °C.

pH:

The pH values in the test medium and the control ranged between 7.3 to 7.5.

Dissolved oxygen:

The oxygen concentration was always 8.3 mg/L or higher.

Salinity:

no data

Nominal and measured concentrations:

nominal: 100 mg/L
measured: 102 % of the nominal value

Details on test conditions:

Light conditions:
A 16-hour light to 8-hour dark photoperiod, with a 30-minute transition period (light intensity during the light period was approximately within the range of 50 to 500 Lux).

Water quality criteria:
The water temperature, pH-values and dissolved oxygen were measured at the start of the test and once every day during the test in the single test concentration and in the control. At the same dates the appearance of the test medium was recorded.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No remarkable observations were made concerning the appearance of the test medium. It was a clear solution colored red by the test item throughout the entire test duration. The pH values in the test medium and the control ranged between 7.3 to 7.5. The oxygen concentration was always 8.3 mg/L or higher, and thus higher than 60 % oxygen saturation. The water temperature was 22 °C.

Reported statistics and error estimates:

The NOEC and the LCO were determined directly from the raw data. The LOEC, the LC100 and the LC50 at the observation times could not be quantified due to the absence of a toxic effect of the test item at the tested concentration.

Validity criteria fulfilled:

yes

Conclusions:

The 96-hour LC50 was higher than 100 mg/L.

Executive summary:

The acute toxicity of the test item FAT 40825/A to zebra fish {Brachydanio rerio) was determined in a 96-hour static test according to the EU Commission Directive 92/69/EEC, Part C.1 (1992), and the OECD Guideline for Testing of Chemicals No. 203, (1992). A limit test was performed in accordance with the guidelines to demonstrate that the test item has no toxic effect on the test organisms at the nominal concentration of 100 mg/L. Thus, the only concentration tested was nominal 100 mg/L and a control. The analytically determined concentration of the test item FAT 40825/A in the test medium was 102 % of the nominal value throughout the test period of 96 hours. The test item FAT 40825/A was stable under the conditions of the test during the test period of 96 hours. Therefore, all reported biological results are related to the nominal concentration of the test item. In the control and at the test concentration of 100 mg/L no mortality or other visible abnormalities were determined at the test fish during the test period of 96 hours. Therefore, the 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), and the 96-hour LCO of FAT 40825/A to zebra fish were determined to be at least 100 mg/L. The NOEC and the LCO might even be higher than this concentration, but concentrations in excess of 100 mg/L have not been tested, according to the guidelines. The 96-hour LOEC (lowest concentration with toxic effects), the 96-hour LC50 and the 96-hour LC100 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 tested concentration.

Description of key information

The 96-hour NOEC and the 96-hour LCO of FAT 40825/A to zebra fish were determined to be at least 100 mg/L. The 96-hour LOEC (lowest concentration with toxic effects), the 96-hour LC50 and the 96-hour LC100 were clearly higher than 100 mg/L, due to the absence of toxicity of FAT 40825/A at the tested concentration.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

> 100 mg/L

Additional information

The acute toxicity of the test item FAT 40825/A to zebra fish (Brachydanio rerio) was determined in a 96-hour static test according to the OECD Guideline for Testing of Chemicals No. 203, (1992). A limit test was performed in accordance with the guidelines to demonstrate that the test item has no toxic effect on the test organisms at the nominal concentration of 100 mg/L. Thus the only concentration tested was nominal 100 mg/L and a control. The analytically determined concentration of the test item FAT 40825/A in the test medium was 102 % of the nominal value throughout the test period of 96 hours. The test item FAT 40825/A was stable under the conditions of the test during the test period of 96 hours. Therefore, all reported biological results are related to the nominal concentration of the test item. In the control and at the test concentration of 100 mg/L no mortality or other visible abnormalities were determined at the test fish during the test period of 96 hours. Therefore, the 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), and the 96-hour LCO of FAT 40825/A to zebra fish were determined to be at least 100 mg/L. The 96-hour LOEC (lowest concentration with toxic effects), the 96-hour LC50 and the 96-hour LC100 were clearly higher than 100 mg/L, due to the absence of toxicity of FAT 40825/A at the tested concentration.