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Environmental fate & pathways

Biodegradation in water: screening tests

Administrative data

biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guidelineopen allclose all
according to guideline
OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
Version / remarks:
adopted 17. Jul. 1992
not specified
according to guideline
EU Method C.4-C (Determination of the "Ready" Biodegradability - Carbon Dioxide Evolution Test)
Version / remarks:
adopted 30. May 2008
not specified
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
1,3,2-dioxathiane 2,2-dioxide
EC Number:
EC Name:
1,3,2-dioxathiane 2,2-dioxide
Cas Number:
Molecular formula:
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material: 1,3,2-dioxathiane 2,2-dioxide
- IUPAC name: 1,3,2-Dioxathiane 2,2-dioxide
- Molecular formula: C3H6O4S
- Molecular weight: 138.14 g/mol
- Substance type: Organic
- Physical state: Solid

Study design

Oxygen conditions:
Inoculum or test system:
activated sludge (adaptation not specified)
Activated sludge from a biologic sewage treatment plant was used as inoculum. The cho-sen plant treats mostly domestic sewage.
Details on inoculum:
- Source of activated sludge (e.g. location, sampling depth, contamination history, procedure): The sludge was taken from the activation basin of the ESN (Stadtentsorgung Neustadt) sewage treatment plant, Im Altenschemel, 67435 NW-Lachen-Speyerdorf.
Date of collection: 28. Sep. 2018, batch no: 20180928.
- Pretreatment: The sludge was filtrated, washed and then aerated until use. And the dry matter was determined.
- Concentration of sludge: The dry matter was determined to contain 4940 mg of suspended solids/L.
- Water filtered: no
- Type and size of filter used, if any: The sludge was filtrated with test medium (2X).
Duration of test (contact time):
28 d
Initial test substance concentration
Initial conc.:
20 other: mg organic carbon/L
Parameter followed for biodegradation estimation
Parameter followed for biodegradation estimation:
CO2 evolution
Details on study design:
- Composition of medium: The medium was freshly prepared. 10 mL of solution a were mixed with 800 mL water, then 1 mL of solutions b, c and d were added and filled up to 1 L with H2O demin (volumes were adapted to final volume needed in the test).
- Test temperature: 20.1 – 22.8 °C without direct lighting
- pH: 7.0-7.6
- pH adjusted: no
- Suspended solids concentration: 4940 mg of suspended solids/L
- Continuous darkness: yes

- Measuring equipment: Analyses of the emitted CO2 were made by IC measurement using the carbon analyser TOC multi N/C 2100S, Analytik Jena. Each sample was measured in duplicate or triplicate, respectively (depending on the variation between the measured values).
- Test vessels: The test vessels were aerated with purified (by activated charcoal), CO2-scrubbed, mois-tened air. 2000 mL-SCHOTT-flasks were used as test vessels, 100 mL scrubber flasks as absorbent vessels.

From each front scrubber flask, 9 samples were taken in order to determine the emitted CO2 (on day 0, 3, 7, 9, 11, 14, 17, 23 and 29). The sample volume was 1 mL. The resulting change in the volume of the front flask was considered in the calculation of emitted CO2 (see also chapter 8.3.1).
On day 28, 5 mL HCl 2 M was added to each test flask in order to drive off dissolved CO2. On day 29, samples from both scrubber flasks were taken.

- Positive control: Aniline (Phenylamine, C6H5NH2, CAS-No. 62-53-3).
Apparatus blanks 2, containing mineral medium only
Blank Controls 2, containing mineral medium and inoculum
Positive control flasks 2, containing positive control, mineral medium and inoculum
Test flasks 2, containing test item, mineral medium and inoculum
Abiotic control 1, containing test item, mineral medium and HgCl2
Toxicity control 1, containing test item, positive control, mineral medium and inoculum
Reference substance
Reference substance:

Results and discussion

% Degradation
Key result
% degradation (CO2 evolution)
Sampling time:
28 d
Details on results:
•The test item 1,3,2-Dioxathiane, 2,2-dioxide is considered as “readily biodegrada-ble“.
•The degree of biodegradation reached 83 % after 28 days.
•The 10-day-window began on day 11, at its end, 63 % degradation were reached, surpassing the pass level of 60 % given in the OECD guideline.
•The abiotic degradation reached 0.4 %.

Applicant's summary and conclusion

Validity criteria fulfilled:
Interpretation of results:
readily biodegradable
The test item 1,3,2-Dioxathiane, 2,2-dioxide was tested using a concentration of nominally 20 mg organic carbon/L 1,3,2-Dioxathiane, 2,2-dioxide in test medium following OECD 301B and EU-Method C.4-C.

Aniline was chosen as positive control.
Activated sludge was used as inoculum (concentration in the test 25.0 mg dry matter/L). The test was left running for 28 days.
All validity criteria were met. Degradation of the positive control surpassed the pass level of 60 % after 10 days.

The following data were determined for the test item 1,3,2-Dioxathiane, 2,2-dioxide:
10-day-window: day 11 – 21
degradation at the end of 10-day-window: 63 %
degradation at the end of the test: 83 %
pass level following guideline: 60 % at the end of 10-day-window for pure substances respective 60 % at the end of the test for mixtures

Therefore, when applying the 10-day-window, 1,3,2-Dioxathiane, 2,2-dioxide is readily biodegradable following OECD 301B and EU C.4-C respectively.
Executive summary:

All validity criteria were met.

Degradation behaviour of positive control and toxicity control was normal. Abiotic degradation reached 0.4 %. Both replicates of the test item showed good correspondence.

If degradation in the toxicity flask is below 25 % after 14 days, the test item can be considered as toxic towards the inoculum. As degradation in the toxicity flask was 61.5 % after 14 days, the test item can be stated as “not toxic towards the inoculum in a concentration of 76.7 mg/L”.

Ready biodegradability is defined in the guidelines as degradation surpassing 60% within 10 days after reaching a level of 10 %. Therefore, the test item1,3,2-Dioxathiane, 2,2-dioxidecan be considered as “readily biodegradable”.

No observations were made which might cause doubts concerning the validity of the study outcome.


The result of the test can be considered valid.