Trisodium 2-[[6-[(4-amino-6-chloro-1,3,5-triazin-2-yl)methylamino]-1-hydroxy-3-sulphonato-2-naphthyl]azo]naphthalene-1,5-disulphonate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline available

Principles of method if other than guideline:

The study report contains the limited information on the test conditions because the study has been performed in 1994. However available information is sufficient to conclude on the classification of the substance. Further the test was performed on the vertebrates and use of results from old experimental studies is one of the options to provide information requested by REACH. New experimental studies with vertebrates must only be conducted if there is no adequate existing information.

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

no

Test organisms (species):

Daphnia magna

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Nominal and measured concentrations:

Nominal concentrations:848.96, 636.72, 424.48, 212.24, 169.79, 106.12, 63.67, 21.22 and 10.61 mg/L

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

177 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

306 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Conclusions:

According to Regulation (EC) No. 1272/2008 (CLP), the test substance, Reactive Orange 13, need not be classified as Aquatic Acute substance.

Reference 2

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

Justification for type of information:

None

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

Principles of method if other than guideline:

None

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL- Source and lot/batch No.of test material: Ref No. 6293 - Expiration date of the lot/batch: 14 October 2019STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL- Storage condition of test material: Frozen in the dark

Analytical monitoring:

yes

Details on sampling:

Samples were taken from the control and 100 mg/L test group from the bulk test preparation at 0 hours and from the pooled replicates at 48 hours for quantitative analysis. All samples were stored frozen prior to analysis. Duplicate samples were taken and stored frozen for further analysis if necessary.

Vehicle:

no

Details on test solutions:

A nominal amount of test item (100 mg) was dissolved in test water and the volume adjusted to1 liter to give the 100 mg/L test concentration.The prepared concentration was inverted several times to ensure adequate mixing and homogeneity.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM - Common name: Daphnia Magna - Source: In-house laboratory cultures - Feeding during testDaphnids were not fed during the main test The test was carried out using 1st instar Daphnia magna derived from in-house laboratory cultures.Adult daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium in a temperature controlled room maintaining the water temperature at 18 to 22°C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

Not applicable

Post exposure observation period:

Not applicable

Hardness:

The reconstituted water had an approximate theoretical total hardness of 250 mg/L as CaCO3.

Test temperature:

Nominal: 18 to 22°C with a maximum deviation of ±1°C.Measured: 22°C (all vessels at each measurement timepoint; 0, 24, and 48 hours).

pH:

The reconstituted water had a pH of 7.8 ± 0.2 adjusted (if necessary) with NaOH or HCl.Measured values: 7.6 - 7.7

Dissolved oxygen:

Measured: 8.0 - 8.8 mg O2/L

Salinity:

Not measured

Conductivity:

Not measured

Nominal and measured concentrations:

Nominal: 0 (control), 100 mg/L.Measured (treated group only; control samples were

Details on test conditions:

TEST SYSTEM - Test vessel: Glass beakers - Type (delete if not applicable): Covered to reduce evaporation - Material, size, headspace, fill volume: 150 mL beaker, 100 mL fill volume - Aeration: No - Type of flow-through: Not applicable - Renewal rate of test solution: Not applicable - No. of organisms per vessel: 5 - No. of vessels per concentration: 4 - No. of vessels per control (replicates): 4 TEST MEDIUM / WATER PARAMETERSReconstituted Water – ISO MediumIngredientFinal Concentration (mg/L)CaCl2.2H2O294MgSO4.7H2O123NaHCO365KCl5.8The reconstituted water had a pH of 7.8 ± 0.2 adjusted (if necessary) with NaOH or HCl and was aerated until the dissolved oxygen concentration was approximately air-saturation value. OTHER TEST CONDITIONS - Adjustment of pH: As noted above, the pH of the reconstituted water was adjusted to pH 7.8±0.2; the pH of the test solutions was not adjusted. - Photoperiod: 16 hours light (200 to 1200 Lux), 8 hours dark, with 20 minute dawn and dusk transition periods. EFFECT PARAMETERS MEASURED: Any immobilization or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure. The criterion of effect used was that daphnia were considered to be immobilized if they were unable to swim within 15 seconds after gentle agitation. VEHICLE CONTROL PERFORMED: Not applicable RANGE-FINDING STUDY - Test concentrations: 0.1, 1.0, 10 and 100 mg/L. - Results used to determine the conditions for the definitive study: Yes (no immobilisation was observed in the 100 mg/L level, so the definitive test was run as a Limit Test at 100 mg/L).

Reference substance (positive control):

yes

Remarks:

A positive control was run as a separate study under similar conditions to those used during the definitive test. Daphnia were exposed to a potassium dichromate control. The date of the positive control study was 29 February to 02 March 2016.

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:

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:

No sub-lethal effects of exposure were observed throughout the test.

Results with reference substance (positive control):

None

Reported statistics and error estimates:

Not applicable.

None

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test item to the freshwater invertebrate Daphnia magna was investigated and gave a 48-Hour EC50 value of greater than 100 mg/L. The No Observed Effect Concentration was 100 mg/L.

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item toDaphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.

Methods

Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to an aqueous solution of the test item at a concentration of 100 mg/L for 48 hours at a temperature of 22°C under static test conditions. Immobilization and any adverse reactions to exposure were recorded after 24 and 48 hours.

 Results…….

Analysis of the test preparations at 0 and 48 hours showed measured test concentrations to range from 99% to 101% of nominal and so the results are based on nominal test concentrations.

Exposure ofDaphnia magnato the test item gave EC₅₀values of greater than 100 mg/L. The No Observed Effect Concentration was 100 mg/L.

It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L