Disodium 4,5-dihydroxybenzene-1,3-disulphonate

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

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Reference 2

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Test guideline 2

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

Test material information

Constituent 1

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 concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

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”.