Reaction products of Sodium 1-amino-4-bromo-9,10-dioxoanthracene-2-sulphonate coupled with Aniline, subsequently coupled with p-(1,1-dimethylpropyl)phenol, subsequently coupled with Ethyl benzoylacetate, subsequently coupled with Fuming Sulfuric acid, sodium salts.

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:

May from 08th to 11th, 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

adopted 13th April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

For the determination of the test item concentration, samples were taken from the test item treated group and from the control group (4 x 5 ml at the start and 4 x 5 ml at the end of the study).

Vehicle:

no

Details on test solutions:

The test solution used in the test was prepared by mechanical dispersion. An amount of 0.0501 g test item was dissolved in 501 ml dilution water (ISO medium) in order to obtain the concentration of 100 mg product/l. The test solution was freshly prepared in the testing laboratory just before introduction of test animals (start of the experiment).

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection), Hungary
- Age of parental stock: less than 24 h old at the beginning of the test.
- Feeding during test: test animals were not fed during the exposure.

ACCLIMATION
- Acclimation period: test animals were bred under similar (or the same) conditions as that used during the exposure period (holding water, temperature, background colour etc.), therefore additional acclimatisation before the test was not necessary. Brood daphnids were maintained in dilution water at the test temperature for at least 48 hours prior to the start of the test.
- Acclimation conditions (same as test or not):
- Type and amount of food: Daphnia were fed with concentrated algal suspension of Raphidocelis subcapitata during the holding.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

20.3 - 20.7 °C

pH:

7.84 - 8.09

Dissolved oxygen:

7.02 - 7.20 mg/l

Nominal and measured concentrations:

100 mg/l (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beaker; volume approx. 50 ml filled with approx.40 ml control or test solution.
- No. of organisms per vessel: 5 animals per replicates.
- 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).
- Theoretical total hardness: 249 mg/l (as CaCO3).

OTHER TEST CONDITIONS
- Photoperiod: 48 h darkness.

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

RANGE-FINDING STUDY
- No. of organisms per vessel: 5 animals per replicates.
- No. of vessels per concentration: 2 replicates
- Test concentrations: 0.1, 1.0, 10.0 and 100.0 mg/l.
- Results used to determine the conditions for the definitive study: only one immobilized daphnid was observed at 100 mg/l after the 48 hours exposure period.

ACCEPTANCE CRITERIA
The immobilisation rate of the introduced Daphnia should not exceed 10 % in the control group.
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/l in control and test vessels.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

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:

No immobilisation or any abnormal behaviour of test animals was observed in the control group and no immobilization or any abnormal behavior of the test animals was observed at the test concentration of 100 mg/l after 48 hours of exposure.

APPEARANCE OF TEST MEDIUM
The test solution was clear solution at 0h (start of experiment) and there was not observed any change of the test solution at 24 h and 48 h.

MEASURED CONCENTRATION
The measured concentration of the test item in the test solution was 94 % of the nominal value at the start and 94 % of the nominal value at the end of the test. There is evidence that the concentration of the tested substance was maintained within ± 20 % of the nominal concentration throughout the test. Therefore the analysis of the results was based on the nominal concentration value. Test item was not detected in the control group.

Immobilization of the test animals

Nominal concentration [mg/l]	Number Daphnia tested	Number of immobilised Daphnia after		% of immobilised Daphnia after
		24 h	48 h	24 h	48 h
Control	20	0	0	0	0
100	20	0	0	0	0

Measured concentrations

Nominal concentration [mg/l]	Mean of the measured concentrations (mg/l)		Measured concentration in % of the nominal
	Start	End	Start	End
100	93.6	94.3	94	94
RSD (%)	2.5	4.5
Control	not detected	not detected

Results of the Preliminary Range-Finding Test

Nominal concentration [mg/l]	Untreated control	0.1	1	10	100
Number of treated animals	10	10	10	10	10
Number of immobilised animals(at 48h)	0	0	0	0	1

Validity criteria fulfilled:

yes

Remarks:

The immobilisation rate of Daphnia was lower than 10 % in the control group and the dissolved oxygen concentration at the end of the test was ≥ 3 mg/l in control and test vessels.

Conclusions:

EC50 (48h) > 100 mg/l (nominal)

Executive summary:

The substance was assessed using Daphnia magna in a 48-hour Acute Immobilisation Test, conducted according to the OECD guideline 202. Based on results obtained in a non-GLP preliminary experiment, daphnids were exposed to a single concentration of 100 mg/l (limit concentration). A concurrent control group was run.

A static test was performed as the test item was previously shown to be stable in the ISO medium for 48 hours. The analytically measured test item concentration remained within ± 20 % of the nominal during the test period of 48 hours; therefore, biological results are based on the nominal concentration.

Twenty Daphnia (divided into 4 replicates) were tested each, exposed to 100 mg/l test item and in the control. The immobilisation of the test animals was recorded 24 and 48 hours after treatment. Environmental conditions were recorded. All measured values remained within acceptable ranges.

No immobilisation or any abnormal behaviour of test animals was observed in the control group and no immobilization or any abnormal behavior of the test animals was observed at the test concentration of 100 mg/l after 48 hours of exposure.

All validity criteria were met and therefore the study can be considered as valid.

Conclusion

EC50 (48h) > 100 mg/l (nominal)

Description of key information

Not harmful/toxic to aquatic invertebrates after short-term exposure (EC50 (48h) > 100 mg/l (nominal))

Additional information

The substance was assessed using Daphnia magna in a 48-hour Acute Immobilisation Test, conducted according to the OECD guideline 202. Based on results obtained in a non-GLP preliminary experiment, daphnids were exposed to a single concentration of 100 mg/l (limit concentration). A concurrent control group was run. A static test was performed as the test item was previously shown to be stable in the ISO medium for 48 hours. The analytically measured test item concentration remained within ± 20 % of the nominal during the test period of 48 hours; therefore, biological results are based on the nominal concentration.

Twenty Daphnia (divided into 4 replicates) were tested each, exposed to 100 mg/l test item and in the control. The immobilisation of the test animals was recorded 24 and 48 hours after treatment. Environmental conditions were recorded. All measured values remained within acceptable ranges.

No immobilisation or any abnormal behaviour of test animals was observed in the control group and no immobilization or any abnormal behavior of the test animals was observed at the test concentration of 100 mg/l after 48 hours of exposure.

All validity criteria were met and therefore the study can be considered as valid.