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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 May - 14 Jun 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guidelineopen allclose all
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
GLP compliance:
yes (incl. QA statement)
Remarks:
Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Mainz, Germany

Test material

Constituent 1
Chemical structure
Reference substance name:
Disodium 4,5-dihydroxybenzene-1,3-disulphonate
EC Number:
205-741-5
EC Name:
Disodium 4,5-dihydroxybenzene-1,3-disulphonate
Cas Number:
149-45-1
Molecular formula:
C6H6O8S2.2Na
IUPAC Name:
disodium 4,5-dihydroxybenzene-1,3-disulphonate

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: Control and 100 mg/L
- Sampling method: Samples were taken at test start and test end

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Differential loading: A stock solution containing 101 mg/L test item in dilution water was prepared and used for the test.
- Controls: Yes, only medium

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Strain/clone: Berlin
- Source: Originally obtained from Bundesumweltamt Berlin, in-house breading since 27. Sep 2007
- Feeding during test: No

ACCLIMATION
- Acclimation conditions: Medium was different M4-medium (husbandry) to dilution water (test)
- Type and amount of food: Green algae (Desmodesmus subspicatus) for stock culture
- Feeding frequency: Twice a week

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: 20 h before test start, the adult animals were separated from the young. 40 min before test start, the adults were caught with the help of a glass tube, and the newborn daphnia, aged between 0 and 19 h and 20 min, were sieved from the medium and immediately placed into a beaker containing dilution water. After the settling-in period, animals which showed no apparent damage were used for the test.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h

Test conditions

Hardness:
2.5 mmol/L and 250 mg CaCO3/L (for dilution water)
Test temperature:
20.3 – 21.8 °C
pH:
7.8 (Control)
7.2 - 7.8 (100 mg/L)
Dissolved oxygen:
8.7 - 8.8 mg/L (Control)
8.7 - 8.8 mg/L (100 mg/L)
Nominal and measured concentrations:
Control and 100 mg/L (nominal)
Control, 119.1 mg/L (based on geomertic mean of each test item concentration)
Details on test conditions:
TEST SYSTEM
Test vessel:
- Type: open (covered with glas lids)
- Material, size, headspace, fill volume: glass beakers, nominal volume 50 mL, tall shape, 20 ± 5 mL test solution
- Aeration: No
- No. of organisms per vessel: 5 organisms
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dilution water, details are given in table 1 in section "any other information on materials and methods incl. tables".
- Culture medium different from test medium: Yes M4-medium (for husbandry) and dilution water (for the test)

OTHER TEST CONDITIONS
- Adjustment of pH: No

EFFECT PARAMETERS MEASURED: Mobility of Daphnia was checked each day

RANGE-FINDING STUDY: No range-finding test was performed
Reference substance (positive control):
yes
Remarks:
Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9)

Results and discussion

Effect concentrationsopen allclose all
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:
EC50
Effect conc.:
> 119.1 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Behavioural abnormalities: No abnormal behaviour was observed
- Mortality of control: No mortality was observed in the control replicates
- Other: Results are summarized within table 1 in section"Any other information on results incl tables"
Results with reference substance (positive control):
Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9) was used as positive control in a current reference study. The 24h-EC50 value was determined as 1.9 mg/L. The value was within the requested range for the 24h-EC50 between 0.6 - 2.1 mg/L of potassium dichromate K2Cr2O7 as specified in the OECD guideline. Since the fits showed sufficient statistical correspondence of the data with the dose-response-equation, this reference study can be considered valid.
Reported statistics and error estimates:
As no toxicity occurred, no statistical evaluation was conducted.

Any other information on results incl. tables

Analytical Results:

At the beginning and at the end of the test, the content of the test item in the test solutions was determined using HPLC-UV-determination. The concentration determined at the start of the test was 116% of the nominal concentration. At the end of the test the determined concentration was 122% of the nominal concentration. Therefore, the determination of the biological results was based on the geometric mean of the measured concentration. However, nominal concentrations were additionally presented.

Table 1: Biological Results Test Item

Parameter

Nominal value

Geometric mean value

48h-NOEC

≥ 100mg/L

≥ 119.1 mg/L

48h-LOEC

> 100 mg/L

> 119.1 mg/L

24h-EC50

> 100 mg/L

> 119.1 mg/L

48h-EC50

> 100 mg/L

> 119.1 mg/L

 

Table 2: Validity criteria

Criterion from the guideline

Outcome

Validity criterion fulfilled

In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.

0

Yes

The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.

8.7 mg/L

Yes

Table 3: Measured Concentrations

Nominal Concentration

Test Item Measured concentration t = 0 h

Measured concentration t = 48 h

% of nominal concentration t = 0 h

% of nominal concentration t = 48 h

mg/L

mg/L

mg/L

mg/L

mg/L

Blank control

< LOQ/LOD

< LOQ/LOD

--

--

100

116.29

122.00

116

122

LOQ (Limit of quantification) = 0.05 mg/L

LOD (Limit of determination) = 0.05 mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
For further details please refer to “Any other information on results incl. tables”.