3,10-diamino-6,13-dichloro-2-((6-(((4-(1,1-dimethylethyl)phenyl)sulfonyl)amino)-2-naphthalenyl)sulfonyl)-4,11-triphenodioxazinedisulfonic acid, lithium potassium sodium salt

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:

2001, September 05 to 2002, March 19

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:

1984

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1992

GLP compliance:

yes

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 BLUE GS 5664.80 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 of the only test concentration of nominal 100 mg/L was prepared by dissolving 30 mg test item in 300 ml test water by intense stirring for 15 minutes at room temperature.
The test media were prepared just before introduction of the daphnids (= start of the test).
The actual concentration of the test item in the test medium was analytically determined.
The test concentration was based on the results of a range-finding test and on results of a pre-experiment to the solubility of the test item (without GLP).
The range-finding test and the pre-experiment to the solubility of the test item were not performed in compliance with GLP-Regulations and therefore are excluded from the Statement of Compliance.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: A clone of this species was originally supplied by the University of Sheffield/UK in 1992, defined from the supplier as clone 5. Since this date the clone is bred in the laboratories of RCC in reconstituted water of the quality identical to the water quality used in the tests (regarding pH, main ions and total hardness) and under temperature and light conditions identical to those of the tests
- Age of parental stock: At the start of the test the used test organisms were 6-24 hours old and were not first brood progeny.
- Feeding during test: none

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/L) as CaC03

Test temperature:

The water temperature was between 20 and 21°C

pH:

The pH value in the control was 7.9 at the start and at the end of the test. The pH value in the test medium ranged from 8.6 at the start to 7.9 at the end of the test.

Dissolved oxygen:

The test water was aerated prior to the start of the study until oxygen saturation was reached. During the test period the test water was not aerated.
At the beginning and the end of the test period, the dissolved oxygen concentrations in the test medium and the control were at least 8.1 mg/L:
- in control: 8.5 mg/L at the start and 8.6 mg/L at the end of the test.
- in the test medium: 8.6 mg/L at the start and 8.3 mg/L at the end of the test.

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 analytically determined test item concentration in the test medium at the start and the end of the test was 105 and 106% of the nominal value, respectively. Under the test conditions the test item BLUE GS 5664.80 was stable during the test period of 48 hours. Therefore all reported results are related to the nominal concentration of the test item.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume:
- Aeration:
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate):
- No. of organisms per vessel:
- No. of vessels per concentration (replicates):
- No. of vessels per control (replicates):
- No. of vessels per vehicle control (replicates):
- Biomass loading rate:

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Reconstituted test water: analytical grade salts were dissolved in purified water to obtain the 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 prior to the start of the study until oxygen saturation was reached. During the test period the test water was not aerated.

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: A 16-hour light to 8-hour darkness photoperiod (with a 30 minute transition period).
- Light intensity: at light period between 200 and 1200 Lux.

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: none
- Appearance of the test medium during the test period: strong coloration caused by the test item (at 0, 24 & 48h). Test item lying at the bottom of the test beaker (at 24 & 48h)

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 LC50 > 100 mg/L

Executive summary:

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, Part I (1984).

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

During the test period the test item concentration in the duplicate samples from the start and the end of the test was measured to be 105 and 106% of the nominal value, respectively. Under the test conditions the test item BLUE GS 5664.80 was stable during the test period of 48 hours. Therefore all reported results are related to the nominal concentration of the test item.

In the control and at the test item concentration of 100 mg/L no immobilized test organisms or other signs of intoxication were determined during the test period of 48 hours.

The 48-hour NOEC {highest concentration tested without toxic effects after the exposure period of 48 hours), and the 48-hour ECO of BLUE GS 5664.80 to Daphnia magna were determined to be at least 100 mg/L. These values might even be higher, but concentrations in excess of 100 mg/L have not been tested, according to the EU Commission Dire.ctive. 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 BLUE GS 5664.80 at the test concentration of 1 00 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L