C.I. Reactive Yellow 2

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:

25 October 2016 to 27 October 2016

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: YAKUSHOKHATSU No. 1121002, SEIKYOKU No.2 and KANPOKIHATSU No. 021121002 and partially amended 2006 as the joint ordinance of The Japanese Ministry of Economy Trade and Industry, Ministry of Health, Labour and Welfare and Ministry of the Environment.

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF 2-7-7.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:

- Concentrations: Analytical measurements were performed at the applied test concentration level (100 mg/L) at the beginning and at the end of the experiment.
- Sampling method: The samples were analyzed by a HPLC-UV method.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method:A stock solution with a concentration of 100 mg/L (nominal) was prepared with direct addition of the test item, mixed into the test medium (ISO Medium). As a limit test was carried out, further dilution of stock solution was not performed.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Justification for species: Daphnia magna is the standard species of the acute immobilisation test.
- Age of parental stock: The Daphnia magna were less than 24 h old at the beginning of the test.
- Feeding during test: The animals were not fed during the test.

ACCLIMATION
- Acclimation period: There was no acclimatization 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:

247 mg/L (as CaCO3)

Test temperature:

19.9 – 20.1 °C

pH:

6.99 – 7.78

Dissolved oxygen:

8.4 – 8.7 mg/L

Nominal and measured concentrations:

- Nominal concentration of 100 mg/L
- Measured concentration was 95.7 mg/L at the start and 99.3 mg/L at the end of the test.
- As the measured concentrations deviated not more than 20 per cent from the nominal, the biological results are based on the nominal concentration.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beaker
- Material, size, headspace, fill volume: At least 5 mL test solution/animal.
- No. of organisms per vessel: Five animals per vessel
- No. of vessels per concentration: 4 replicates
- No. of vessels per control: 4 replicates

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. Separate stock solutions of individual trace elements were first prepared in deionised water. The medium was prepared of these different stock solutions, so it contained all trace elements (combined solution). See Table 1 for the composition of ISO medium.

OTHER TEST CONDITIONS
- Adjustment of pH: The pH of the test solution was not adjusted and not varied by more than 1.5 units in any one test.
- Photoperiod: The test was carried out in 16-hour light and 8-hour dark cycle.

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.
The water temperature was measured daily; the oxygen concentrations and pH of the controls and the test solution were measured at the beginning and at the end of the experiment.

RANGE-FINDING STUDY
A concentration range-finding test was conducted to determine the approximate toxicity of the test item so that appropriate test concentrations could be selected for use in the definitive test. Ten daphnids (divided into two replicates) in each test concentration and control were exposed for 48 hours under semi-static conditions (in absence of stability data).
The test solutions were prepared by appropriate dilution of the stock solution.
- Test concentrations: The range-finding test was conducted at nominal test substance concentrations of 0.10, 1.0, 10 and 100 % saturated solution under static test conditions.
- Results used to determine the conditions for the definitive study: The choice of the test concentration was made on the basis of the results of a preliminary range-finding test. Because toxic response was not observed during the preliminary concentration rangefinding 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 is tested at least twice a year to demonstrate satisfactory test conditions.

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:

EC100

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

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Immobilisation: The number of immobilised animals and the percentage of immobility were determined at the 24th and 48th hour. None of the daphnia were immobilised during the test.
- Abnormal responses: No abnormal behaviour or appearance of test animals was detected.
- Mortality of control: There was no immobilization in the control group.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
The 24h EC50 of the reference item, Potassium dichromate, was 0.93 mg/L (95 % confidence limits: 0.86 – 0.99 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

There was no immobilisation in the control group and the dissolved oxygen concentration at the end of the test in control and test vessels was no more than 3 mg/L.

All validity criteria were within acceptable limits and therefore the study can be considered as valid.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the study, the 48h EC50 was >100 mg/L, the 48h EC100 was >100 mg/L, the 48h NOEC was determined as 100 mg/L and the 48h LOEC was determined as 100 mg/L based on nominal concentrations.

Executive summary:

The toxicity of the test material to the freshwater invertebrate, Daphnia magna, was investigated in a study which was conducted under GLP conditions and in accordance with the standardised guideline OECD 202, EU Method C.2 and and the Japanese guidelines YAKUSHOKHATSU No. 1121002, SEIKYOKU No.2 and KANPOKIHATSU No. 021121002 and JMAFF 2-7-7.

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.

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

The test concentration was analytically determined at the start and at the end of the experiment. Measured concentration was 95.7 mg/L at the start and 99.3 mg/L at the end of the test.

As the measured concentrations deviated not more than 20 per cent from the nominal in all cases, biological results are based on the nominal concentration.

Twenty animals, divided into four groups (glass beaker) of five animals each were used at the test concentration and for the control. All validity criteria were met during this study.

Under the conditions of the study, the 48h EC50  was >100 mg/L, the 48h EC100  was >100 mg/L, the 48h NOEC was determined as 100 mg/L and the 48h LOEC was determined as 100 mg/L based on nominal concentrations.

Description of key information

Under the conditions of the study, the 48h EC50  was >100 mg/L, the 48h EC100  was >100 mg/L, the 48h NOEC was determined as 100 mg/L and the 48h LOEC was determined as 100 mg/L based on nominal concentrations.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The toxicity of the test material to the freshwater invertebrate, Daphnia magna, was investigated in a study which was conducted under GLP conditions and in accordance with the standardised guideline OECD 202, EU Method C.2 and and the Japanese guidelines YAKUSHOKHATSU No. 1121002, SEIKYOKU No.2 and KANPOKIHATSU No. 021121002 and JMAFF 2-7-7. The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

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.

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

The test concentration was analytically determined at the start and at the end of the experiment. Measured concentration was 95.7 mg/L at the start and 99.3 mg/L at the end of the test.

As the measured concentrations deviated not more than 20 per cent from the nominal in all cases, biological results are based on the nominal concentration.

Twenty animals, divided into four groups (glass beaker) of five animals each were used at the test concentration and for the control. All validity criteria were met during this study.

Under the conditions of the study, the 48h EC50  was >100 mg/L, the 48h EC100  was >100 mg/L, the 48h NOEC was determined as 100 mg/L and the 48h LOEC was determined as 100 mg/L based on nominal concentrations.