Phenol, 2,4-dinitro-, sulfurized, thiosulfonated

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:

2019-04-30 to 2019-05-02

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)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2016

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100 mg/L
- Sampling method: For the determination of the test item concentration, samples were taken from the test item treated group and from the control group (4 x 10 mL at the start and 4 x 10 mL at the end of the study).

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION:
The test solution used in the test was prepared by mechanical dispersion without using any solubilizing agent. An amount of 0.0314 g test item was dissolved in 314 mL dilution water (ISO medium see 5.4) in order to obtain the nominal concentration of 100 mg/L. Thereafter this solution was placed into an ultrasonic bath for approximately 5 minutes. The test solution was freshly prepared in the testing laboratory just before introduction of test animals (start of the experiment). No precipitation or particles were observed.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (Straus)
- Strain/clone: Straus
- Age at study initiation: Less than 24 h old at the beginning of the test.
- Sex: female
- Method of breeding: The Daphnia are bred in the Laboratory of TOXI-COOP ZRT.
- Source: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection) 2100 Gödöllő, Kotlán S. u. 3. Hungary
- Feeding during test :Test animals were not fed during the exposure.


ACCLIMATION
- Acclimation period and conditions: 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.
- Food and Feeding: The Daphnia were fed with concentrated algal suspension of Raphidocelis subcapitata during the holding.
- Health during acclimation (any mortality observed): Apparently healthy animals were used in this test with a known history

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

249 mg/L (as CaCO3)

Test temperature:

21.0 – 21.3 °C (in the test vessels)
20.3 – 21.4 °C (within the climate chamber)

pH:

7.29 - 8.10

Dissolved oxygen:

8.03 –8.38 mg/L

Salinity:

Not applicable.

Conductivity:

No data.

Nominal and measured concentrations:

Nominal concentrations: 100 mg/L
Measured concentrations: 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.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beaker
- Material, size, fill volume: glass, volume approx. 50 mL filled with approx. 40 mL
- Aeration: No
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: According to OECD guideline 202

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light: 8 h dark
- Light intensity: The measured light intensity was 749 lux in the climate chamber at the start of the test.

EFFECT PARAMETERS MEASURED: Determination of 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 are considered to be immobile. In addition to immobility any abnormal behaviour or appearance is reported.

RANGE-FINDING STUDY
- Test concentrations: 100 mg/L
- Results used to determine the conditions for the definitive study: In the preliminary test ten daphnids (divided into 2 replicates) were exposed to the test item at a concentration of 100 mg/L for 48 hours. The test solution used in the test was prepared by mechanical dispersion. 0.0306 g of test item was dissolved in 306 mL dilution water (ISO medium) in order to obtain the concentration of 100 mg/L. This test solution was placed into an ultrasonic bath for approximately 5 minutes. Based on the results of the preliminary test, the test item had no toxic effect on the daphnids up to a concentration of 100 mg/L. Based on the results of the non-GLP Preliminary Range-Finding Test, a single nominal concentration of 100 mg/L (limit test concentration) and a concurrent control were tested in the main study.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks:

mortality

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks:

mortality

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks:

mortality

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.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- 24h EC50: 1.58 mg/L (95% conf. limits: 1.34 – 1.85 mg/L)

Reported statistics and error estimates:

A limit test was performed and no toxic effects were observed, therefore statistical analysis was not necessary. The NOEC and LOEC values of the test item were determined directly from the raw data.

Analytical results

The quantification of the test item in the test solutions was performed by a previously validated analytical method (Study Code: 805-100-4366) by the analytical laboratory of TOXI-COOP ZRT. Samples were taken from the test concentration and the control at the start and at the end of the experiment and analysed by VIS spectrophotometric method.

The measured concentration of the test item in the test solution was 96 % of the nominal value at the start and 100 % 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.

 

Nominalconcentrationmg/L	Start (April 30, 2019)		End (May 02, 2019)
	Mean of the measured concentrations (mg/L) (n=4)	Measured concentration in % of the nominal	Mean of the measured concentrations (mg/L) (n=4)	Measured concentration in % of the nominal
100	95.6±0.70	96	100.4±0.47	100
Control	<LOD	-	<LOD	-

 

Validity criteria fulfilled:

yes

Conclusions:

In this acute immobilisation study with Daphnia magna according to OECD guideline 202, the 48-h NOEC was determined to be 100 mg/L and the 48-h LOEC value was determined to be >100 mg/L (corresponding to 16 and >16 mg/ dyestuff/L, repsectively).

Executive summary:

The influence of the test item Solubilized Sulphur Black 1 liquid on the mobility respectively survival of Daphnia magna was assessed in an acute immobilisation test according to OECD guideline 202. Young Daphnia were exposed to aqueous test media containing the test item for 48 hours. Based on results obtained in a non-GLP preliminary experiment Daphnia were exposed to a single concentration of 100 mg/L (limit concentration), corresponding to 16 mg dyestuff/L. 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 quantification of the test item was performed by a previously validated analytical method. Samples were taken from the test concentration and the control at the start and at the end of the test. 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. 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 (equivalent to 16 mg dyestuff/L). The 48 -h NOEC was determined to be 100 mg/L and the 48 -h LOEC value was determined to be > 100 mg/L (corresponding to >16 mg/dyestuff/L). All validity criteria were met and therefore the study can be considered as valid.

Description of key information

In an acute immobilisation study with Daphnia magna according to OECD guideline 202, no immobilization or any abnormal behavior of the test animals was observed at the limit concentration of 100 mg/L (equivalent to 16 mg dyestuff/L). The 48 -h NOEC was determined to be 100 mg/L and the 48-h LOEC value was determined to be > 100 mg/L.

Additional information

The influence of the test item Solubilized Sulphur Black 1 liquid on the mobility respectively survival of Daphnia magna was assessed in an acute immobilisation test according to OECD guideline 202. Young Daphnia were exposed to aqueous test media containing the test item for 48 hours. Based on results obtained in a non-GLP preliminary experiment Daphnia were exposed to a single concentration of 100 mg/L (limit concentration, corresponding to 16 mg dyestuff/L). 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 quantification of the test item was performed by a previously validated analytical method. Samples were taken from the test concentration and the control at the start and at the end of the test. 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. 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. The 48-h NOEC was determined to be 100 mg/L and the 48-h LOEC value was determined to be > 100 mg/L. All validity criteria were met and therefore the study can be considered as valid