disodium 7-(4,6-difluoropyrimidin-2-ylamino)-4-hydroxy-3-(4-methoxy-2-sulfonatophenylazo)naphthalene-2-sulfonate; reaction mass of: disodium 7-(2,4-difluoropyrimidin-6-ylamino)-4-hydroxy-3-(4-methoxy-2-sulfonatophenylazo)naphthalene-2-sulfonate

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:

Start of Experiments: November 12, 1996 - Completion of Experiments: December 19, 1996

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)

Version / remarks:

adopted April 04, 1984

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

dated December 29, 1992

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

For the analysis of the actual test item concentrations the following samples were taken:
Just before test start:
- duplicate samples from the test medium (without daphnids)
- duplicate samples from the control (without daphnids}

After 48 hours (stability samples) :
- duplicate samples from the test medium
- duplicate samples from the control

For the 48-hour stability samples the contents of the two respective test beakers were combined.
The 0-hour 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 is sufficiently stable in the test water under these storage conditions. The 48-hour samples were analyzed immediately after sampling.
The concentration of the test item SCARLET RN 1165 was analyzed 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 analyzed from each of both sampling times.

Vehicle:

no

Details on test solutions:

The test medium with the test substance concentration of nominal 100 mg/l was prepared by dissolving the test substance homogeneously in test water by intense stirring. The test medium was freshly prepared just before the start of the test(= introduction of Daphnia).

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: supplied by the Ruhrverband, Essen, F.R.G. in 1989
- Feeding during test:
- Food type:
- Amount:
- Frequency:

ACCLIMATION
- Acclimation conditions (same as test or not): The clone was bred in the laboratories of RCC under similar temperature and light conditions as in the test, and in reconstituted water of a similar quality regarding to pH, components of the main ions and total hardness as the test water used in the test



METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:

Test type:

static

Water media type:

other: Reconstituted test water

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

None

Hardness:

2.5 mmol/l (= 250.0 mg/I) as CaC03.

Test temperature:

20.7 - 20.8°C during the test period.

pH:

During the test period, the pH-values ranged from pH 7.7 to pH 8.0.

Dissolved oxygen:

During the test period, the dissolved oxygen concentrations in the test media were at least 8.0 mg/l or higher.

Salinity:

CaCl2 x2H20 : 2.0 mmol/L (= 294.0 mg/L)
MgS04 x 7H20 : 0.5 mmol/L (= 123.0 mg/L)
NaHC03 : 0.75 mmol/L (= 65.0 mg/L)
KCI : 0.075 mmol/L (= 5.8 mg/L)

Conductivity:

Not specified

Nominal and measured concentrations:

Nominal: 100 mg/L
The analytical determined test substance concentrations in the test medium of nominal 100 mg test substance/I varied in the range of 91 % to 94 % of the nominal value during the test period. The mean measured concentration (calculated as the average over all measurements per concentration) was 92 % of nominal.
Therefore, all reported results are related to the nominal concentration of the test substance.

Details on test conditions:

TEST SYSTEM
In each treatment, the test concentration and the control, 20 Daphnia were tested, divided into four groups of five animals, each group in 50 ml test medium in a 100-ml-glassbeaker.
The test was performed in an air-temperated room.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Reconstituted water: in deionized water analytical grade salts were added to following nominal concentrations
CaCl2 x2H20 : 2.0 mmol/L (= 294.0 mg/L)
MgS04 x 7H20 : 0.5 mmol/L (= 123.0 mg/L)
NaHC03 : 0.75 mmol/L (= 65.0 mg/L)
KCI : 0.075 mmol/L (= 5.8 mg/L)
Water Hardness : 2.5 mmol/L (=250.0 mg/L) as CaC03
Alkalinity : 0.8 mmol/L
Ratio of Ca : Mg = 4 : 1 (based on molarity)
Na : K = 1 O : 1 (based on molarity)

The test water was aerated to oxygen saturation.

OTHER TEST CONDITIONS
- Photoperiod: A 16-hour light to 8-hour darkness photoperiod
- Light intensity: light intensity at light period approximatively 400-550 Lux

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Key result

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

In the control and in the test concentration of nominal 100 mg test substance/I no immobility or mortality of the test animals was 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 ECO of SCARLET RN 1165 to Daphnia magna were determined to be at least 100 mg test substance/I. The NOEC and the EC0 might even be higher than this concentration, but concentrations in excess of 100 mg test substance/I have not been tested according to the EEC Commission Directive. The 48-hour LOEC (lowest concentration with toxic effects) and the 48-hour EC 50 were clearly higher than 100 mg test substance/l. These values could not be quantified due to the absence of toxicity of SCARLET RN 1165 up to this test concentration.

Results with reference substance (positive control):

Positive control not used.

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test item BLUE GS 5664.80 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, (1984), resulting in a 48-hour EC50 > 100 mg/L.

Executive summary:

The acute toxicity of the test substance SCARLET RN 1165 to Daphnia magna was determined in a 48-hour static test according to the Commission Directive 92/69/EEC, Annex Part C.2, dated December 29, 1992, and the OECD Guideline for Testing of Chemicals, Section 2, No. 202, Part I, adopted April 04, 1984.

A limit test was performed in accordance with the Commission Directive 92/69/EEC to demonstrate that the test substance has no toxic effect on the test animals up to the concentration of nominal 100 mg test substance/l. Thus the only concentration tested was nominal 100 mg test substance/l and a control.

The analytical determined test substance concentrations in the test medium of nominal 100 mg test substance/l varied in the range of 91 % to 94 % of the nominal value during the test period. The mean measured concentration (calculated as the average over all measurements per concentration) was 92 % of nominal. Therefore, all reported results are related to the nominal concentration of the test substance.

The 48-hour NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours), and the 48-hour EC0 of SCARLET RN 1165 to Daphnia magna were determined to be at least 100 mg test substance/I. The NOEC and the EC0 might even be higher than this concentration, but concentrations in excess of 100 mg test substance/l have not been tested. The 48-hour LOEC (lowest concentration with toxic effects) and the 48- hour EC 50 were clearly higher than 100 mg test substance/I. These values could not be quantified due to the absence of toxicity of SCARLET RN 1165 up to this test concentration.

Description of key information

The acute toxicity of the test item BLUE GS 5664.80 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, (1984), resulting in a 48-hour EC50 > 100 mg/L.