2-[[4-chloro-6-[[8-hydroxy-3,6-disulpho-7-[(1-sulpho-2-naphthyl)azo]-1-naphthyl]amino]-1,3,5-triazin-2-yl]amino]-5-sulphobenzoic acid, 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:

16 October 2017 to 18 October 2017

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF 2-7-2-1 The guidelines related to the study reports for the registration application of pesticide (Ref. No. 12-Nousan-8147 on 24 November 2000) & Ref. No.13-Seisan-3986 on 10 October 2001.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Analytical measurements were performed from the control and at the applied test concentration level at the beginning and at the end of the test.

Vehicle:

no

Details on test solutions:

- Test Material
A stock solution with a concentration of 100 mg/L (nominal) was prepared with the test material and test medium (ISO Medium). As a limit test was carried out, further dilution of stock solution was not performed. The colour of the stock solution was red.

- Untreated Control
The dilution water (ISO-medium) was used without of addition of the test material.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: Szent István University, Department of Aquaculture 2100 Gödöllő, Páter Károly u. 1. – Hungary.
- Age: organisms were less than 24 h old at the beginning of the test
- Feeding during test: no

ACCLIMATION
There was no acclimatisation because the water used was similar to the culture water.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

The reconstituted water (ISO medium) had a total hardness of 247 mg/L (as CaCO3).

Test temperature:

20.1 - 20.6°C (test vessels); 19.8 - 20.9°C (climate chamber)

pH:

7.04 - 7.35

Dissolved oxygen:

8.2 - 8.5 mg/L

Nominal and measured concentrations:

100 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beaker
- Fill volume: at least 5 mL test solution/animal
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water (ISO medium, according to OECD 202) was used as dilution water for both the range finding and definitive tests. The same composition of reconstituted water was used for the tests and for breeding the test animals.
- Intervals of water quality measurement: The water temperature was measured daily, the oxygen concentrations and pH of the control and the test solution were measured at the beginning and at the end of the experiment.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light / 8 hours dark

EFFECT PARAMETERS MEASURED
The immobility or mortality of the Daphnia was determined by visual observation 24 and 48 hours after the start of the test. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker are considered to be immobile.
The number of immobilised animals and the percentage of immobility were determined at 24 and 48 hours.

RANGE-FINDING STUDY
- Test concentrations: 0.1, 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: yes
Because a toxic response was not observed during the preliminary concentration range-finding test, a limit test was carried out using only one test concentration (100 mg/L) and one control group in a static system.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

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

Details on results:

None of the organisms appeared to be immobilised on either the control group or the test material treated group at any observation point. In addition to immobility, no abnormal behaviour or appearance of test animals was detected.

Results with reference substance (positive control):

The 24h EC50: 0.64 mg/L, (95 % confidence limits: 0.59 – 0.68 mg/L).

Reported statistics and error estimates:

No statistical analysis was performed because of the lack of toxic effects.
The EC50, NOEC, LOEC and EC100 values were determined directly from the raw data.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the study the 48 hour EC50 was determined to be > 100 mg/L.

Executive summary:

Acute toxicity of the test material on Daphnia magna was assessed in an Acute immobilisation test, over an exposure period of 48 hours in a static test system. The study was conducted under GLP conditions and in accordance with the standardised guidelines OECD 202, EU Method C.2, US EPA OCSPP 850.1010 and JMAFF 2-7-2-1.

Because no toxic response was observed during the preliminary range-finding test, only a limit test was carried out using only one test concentration (100 mg/L) and one control group in the definitive study.

The test concentration was analytically determined at the start and at the end of the experiment. As the measured concentrations deviated not more than 20 per cent from the nominal, biological results are based on the nominal concentration.

Twenty animals, divided into four replicates (glass beaker) of five animals each were used at the test concentration and for the control.

No statistical analysis was performed because of the lack of toxic effects. The EC50, NOEC, LOEC and EC100 values were determined directly from the raw data. All validity criteria were met during this study.

Under the conditions of the study the 48 hour EC50 was determined to be > 100 mg/L.

Description of key information

Under the conditions of the study the 48 hour EC50 was determined to be > 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

Acute toxicity of the test material on Daphnia magna was assessed in an Acute immobilisation test, over an exposure period of 48 hours in a static test system. The study was conducted under GLP conditions and in accordance with the standardised guidelines OECD 202, EU Method C.2, US EPA OCSPP 850.1010 and JMAFF 2-7-2-1. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Because no toxic response was observed during the preliminary range-finding test, only a limit test was carried out using only one test concentration (100 mg/L) and one control group in the definitive study.

The test concentration was analytically determined at the start and at the end of the experiment. As the measured concentrations deviated not more than 20 per cent from the nominal, biological results are based on the nominal concentration.

Twenty animals, divided into four replicates (glass beaker) of five animals each were used at the test concentration and for the control.

No statistical analysis was performed because of the lack of toxic effects. The EC50, NOEC, LOEC and EC100 values were determined directly from the raw data. All validity criteria were met during this study.

Under the conditions of the study the 48 hour EC50 was determined to be > 100 mg/L.