Tetrasodium [3-[(3-chloro-2-hydroxy-5-sulphophenyl)azo]-5-[[4-chloro-6-(3-sulphoanilino)-1,3,5-triazin-2-yl]amino]-4-hydroxynaphthalene-2,7-disulphonato(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:

2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

published in O.J. L 142, 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

at beginning and end of the test

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM - Common name: Daphnia magna - Strain/clone: Straus - Source: own laboratory - Age of parental stock (mean and range, SD): 24 h

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

no

Hardness:

2.254 mmol/l

Test temperature:

20 ± 2°C with variation ± 1°C

pH:

7.5 - 7.8

Dissolved oxygen:

8.2 - 8.7 mg/l

Nominal and measured concentrations:

nominal concentration: 1 mg/l- 0 h: 1.01 mg/l- 48 h: 1.28 mg/lnominal concentration: 100 mg/l- 0 h: 92.4 mg/l- 48 h: 95.5 mg/l

Details on test conditions:

DILUTION WATERStandard dilution water was prepared just before the test by admixing 2.5 mL of stored solutions A, B, C and D in 1000 mL of deionized water.A. 117.6 g CaCl2·2H2O in 1L deionized waterB. 49.3 g MgSO4·7H2O in 1L deionized waterC. 25.9 g NaHCO3 in 1L deionized waterD. 2.3 g KCl in 1L deionized waterThe dilution water should have a pH 7.8 ± 0.2, values measured in this study were 7.8 in the preliminary test. The content of sum Ca + Mg ions should have the value of 2.50 ± 0.10 mmol/l, values measured in this study were 2.54 mmol/l in the preliminary test. Measured values meet the requirements.INSTRUMENTS AND EQUIPMENT - oximeter WTW Oxi 730 with membrane probe CellOx 325- pH meter WTW pH 539 - conductivity meter FE30 (Mettler Toledo)- analytical balance AG 135 (Mettler Toledo)- equipment for keeping and checking of temperature in test area- beakers, volumetric flasks, pipettesSTUDY CONDITIONSTemperature: 22 ± 2°C, with a variation of ± 1°C at each individual testExposure time: 48 hoursLighting: daylightNumber of animals at one concentration level: 20 for each concentration and controlVolume solution: 50 mL (at least 2 mL per individual)Concentration of dissolved oxygen: higher than 3 mg/LDesign of test: static testWithout feeding, without aeration.PREPARATION OF SOLUTIONS OF THE TEST SUBSTANCEThe stock solution of the test substance was prepared in the dilution water. 50 mg of the test substance was weighed into 500 ml of the dilution water for the preliminary test. The concentrations of solutions used in the preliminary test were obtained by dilution of the stock solution with dilution water.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 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:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No immobilisation found at concentration 1 - 100 mg/l.

Results with reference substance (positive control):

Substance for positive control: potassium dichromateEC50 = 1.17 mg/l (24 h)EC50 = 0.62 mg/l (48 h)

Table No. 1: Immobilisation of daphnia

nominal concentration (mg/l)		100	50	5	1	control
number of mobile animals	24 h / 1st series	10	10	10	10	10
	24 h / 2nd series	10	10	10	10	10
	24 h / total	20	20	20	20	20
immobilisation (%)	24 h	0	0	0	0	0
number of mobile animals	48 h / 1st series	10	10	10	10	10
	48 h / 2nd series	10	10	10	10	10
	48 h / total	20	20	20	20	20
immobilisation (%)	48 h	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

The experiment with the concentration of 100 mg/l included in the preliminary test has been performed using 20 daphnia, therethrough the conditions of the limit test, given in the guideline, were fulfilled. It was demonstrated that the EC50 is higher than this concentration.

Executive summary:

The test substance, Reactive Violet 1, was tested in acute immobilisation test on Daphnia magna. The test was performed according to method C.2 – Daphnia sp. Acute Immobilisation Test, Council Regulation (EC) No. 440/2008, published in O.J. L 142, 2008.

The preliminary test was performed in a range of the test substance nominal concentrations 1 – 100 mg/l. The test was performed as static. Based on no toxicity of the test substance found in preliminary test, the study was terminated. The experiment with the concentration of 100 mg/l included in the preliminary test has been performed using 20 daphnia, therethrough the conditions of the limit test, given in the guideline, were fulfilled. It was demonstrated that the EC₅₀is higher than this concentration.

The samples for analytical determination were taken at the beginning and at the end of the test. The analytical results from the preliminary test showed that the test substance, Relative Violet 1, was in the highest concentration (100 mg/l) stable in the dilution water at conditions of the test.

Test results:

EC₅₀> 100 mg/l (24 h)

EC₅₀> 100 mg/l (48 h)

EC₀> 100 mg/l (24 h)

EC₀> 100 mg/l (48 h)

EC₁₀₀> 100 mg/l (24 h)

EC₁₀₀> 100 mg/l (48 h)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L