Reaction mass of 2,2'-thiodiethanethiol and 1095071-01-4 and α-sulfanyl-ω-{2-[(2-sulfanylethyl)sulfanyl]ethyl}bis[ethane- 1,2-diylsulfanediylethane- 1,2-diylsulfanediyl(3-hydroxypropane-1,1-diyl) sulfanediyl]

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

13 April 2004

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 2,2'-thiodiethanethiol and 1095071-01-4 and α-sulfanyl-ω-{2-[(2-sulfanylethyl)sulfanyl]ethyl}bis[ethane- 1,2-diylsulfanediylethane- 1,2-diylsulfanediyl(3-hydroxypropane-1,1-diyl) sulfanediyl]

EC Number:

947-988-8

Molecular formula:

not applicable

IUPAC Name:

Reaction mass of 2,2'-thiodiethanethiol and 1095071-01-4 and α-sulfanyl-ω-{2-[(2-sulfanylethyl)sulfanyl]ethyl}bis[ethane- 1,2-diylsulfanediylethane- 1,2-diylsulfanediyl(3-hydroxypropane-1,1-diyl) sulfanediyl]

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

yes

Remarks:

HPLC

Details on sampling:

- Concentrations: nominal test concentrations of 8.0, 3.64, 1.66, 0.74 and 0.34 mg/L (factor 2.2) and a control were tested
- Sampling method: Duplicate samples from the freshly prepared test media of all test concentrations and the control were taken at the start of the test.
For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the test period, duplicate samples from the test media of all test concentrations and the control were collected at the end of the test (after 48 hours) by pouring together the contents of the test beakers of each treatment.
All samples were diluted by a factor of two with acetonitrile.
Additional samples of the control and the dilution solvent acetonitrile were taken at each sampling without any sample treatment.
- Sample storage conditions before analysis: All samples were stored in a freezer (≤ - 20 °C), protected from light, until analysis was performed. Afterwards the samples were again stored deep frozen (≤ -20 °C) and will be kept stored up to the date of the final report.

Test solutions

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test item is not well soluble in test medium. To avoid physical effects of undissolved test item on the daphnids, no concentrations above the solubility limit of the test item in test water was tested.
Therefore, a supersaturated stock solution of nominal 8 mg test item/L was prepared by suspending 12.3 mg test item in 1537.5 mL test water. The stock suspension was stirred for 24 hours at room temperature in the dark to dissolve as much test item as possible.
Then, non-dissolved fractions of the test item were separated from the test medium by membrane filtrations (0.65 µm and 0.45 µm cellulose acetate filter). The solution with dissolved test item was used as the test medium of the highest test concentration and to prepare the desired 1:2.2, 1:4.8, 1:10.8 and 1:23.5 dilutions.
The test media were prepared just before introduction of the daphnids (= start of the test).
- Test concentration separation factor: 2.2
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): There were no remarkable observations, except for the stock solution, where particles were swimming at the surface before filtration.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: Daphnia magna (Straus), clone 5
- Age at study initiation (mean and range, SD): From 0.5 to 22.0 hours old
- Method of breeding: The daphnids were bred in the laboratories of ibacon under similar temperature and light conditions as used in the test. The cultivation of the parental daphnids was performed in Elendt M4 medium. The test organisms were not first brood progeny. The daphnids in the stock culture were fed at least on all working days with green algae (Desmodesmus subspicatus) freshly grown in the laboratories of ibacon.
- Feeding during test: none

ACCLIMATION
- Acclimation period: Was not necessary, since the test was performed in the same medium as the culturing
- Acclimation conditions (same as test or not): as test

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

2.5 mmol/L (= 250 mg/L) as CaCO 3

Test temperature:

20.6 to 20.7 °C at test start;
20.1 to 20.6 °C at test end

pH:

7.9 to 8.2 at test start;
7.9 at test end; and thus the pH-value did not vary by more than 1.5 units

Dissolved oxygen:

8.9 to 9.1 mg/L at test start;
8.6 to 8.8 mg/L at test end

Conductivity:

< 5 µScm -1

Nominal and measured concentrations:

Nominal: 8.00, 3.64, 1.66, 0.74, and 0.34 mg/L
arithmetic mean measured: 4.00, 2.02, 0.896, 0.443 and 0.125 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers
- The test vessels were covered with a lid to reduce the loss of water due to evaporation and to avoid the entry of dust into the solutions.
- Material, size, headspace, fill volume: Glass, 100 mL, fill volume: 60 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: Reconstituted Water (Elendt "M4")
- Alkalinity: 0.9 mmol/L
- Conductivity: < 5 µScm -1
- Culture medium different from test medium: no


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8
- Light intensity: 410 to 830 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The mobility of the daphnids was determined by visual observation after 24 and 48 hours. Those animals that are not able to swim within 15 seconds after gentle agitation of the test beaker were considered to be immobile (even if they could still move their antennae).

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Test concentrations:
- Results used to determine the conditions for the definitive study:

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 4 mg/L

95% CI:

>= 3.12 - <= 4

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 4 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Any other information on results incl. tables

Mean measured concentration [mg test item/L]	No. of Daphnia tested	No. of immobilized Daphnia after		% of immobilised Daphnia after
		24 h	48 h	24 h	48 h
Control	20	0	0	0	0
0.125	20	0	0	0	0
0.443	20	0	0	0	0
0.896	20	0	0	0	0
2.02	20	0	2	0	10
4.00	20	0	6	0	30

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The toxic effect of the test item HIDT to Daphnia magna was assessed in a static concentration-response test. The 48-hour NOEC was determined to be 0.896 mg test item/L. The 48-hour LOEC was determined to be 2.02 mg test item/L and the 48-hour EC50 value was determined to be > 4.00 mg test item/L. The initial concentrations and the maintenance of the exposure concentrations during the test were verified in the analytical part. All reported results refer to mean measured values since a filtrate was used and the concentrations of the test item were not within ± 20% of the nominal concentrations during the test.

Executive summary:

The 48 –hr-acute toxicity of 1-propanol, 3,3-bis[[2-[(2-mercaptoethyl)thio]ethyl]thio]- to Daphnia magna was studied under static conditions. Test species were exposed to control and test chemical at (nominal/measured) concentration of 0.34, 0.74, 1.66, 3.64, and 8.00 mg a.i./L and 0.125, 0.443, 0.896, 2.02, and 4.00 mg a.i./L for 48 hr. Immobilisation and sublethal effects were observed daily. The 48 – h EC50 was > 4.00 mg a.i./L (> 8.00 mg a.i/L nominal).  The 48 – h NOEC based on immobilization was 0.896 mg a.i./L (1.66 mg/L nominal). 

Based on the results of this study, {test material} would be not classified as acutely toxic to Daphnia magna in accordance with Regulation (EU) No. 1272/2008 (CLP).

Results Synopsis

 

Test Type (Static):

 

EC50:  > 4.00 mg a.i./L                            95% C.I. :  {3.12 to 4.00 mg a.i./L}

EC50:  > 8.00 mg a.i./L                            nominal

NOEC:  0.896 mg a.i./L}                       

NOEC:  1.66 mg a.i./L}                            nominal

Endpoint(s) Effected: Mobility