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Environmental fate & pathways

Biodegradation in water: screening tests

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Administrative data

Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-03-02 to 2012-07-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
Version / remarks:
July 1992
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Biphenyl-2-yl-(9,9-dimethyl-9H-fluoren-2-yl)-(9,9’-spirobifluoren-2-yl)-amine
EC Number:
700-681-0
Cas Number:
1364603-07-5
Molecular formula:
C52H37N
IUPAC Name:
Biphenyl-2-yl-(9,9-dimethyl-9H-fluoren-2-yl)-(9,9’-spirobifluoren-2-yl)-amine
Test material form:
solid
Specific details on test material used for the study:
Batch: OS11008922
Appearance: pale yellow powder without visible impurities
Released until: August 15, 2016

Study design

Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, non-adapted
Details on inoculum:
- Source of inoculum/activated sludge: Municipal sewage treatment plant, D-31137 Hildesheim

- Pretreatment:
The activated sludge was washed twice with autoclaved tap water. After the second washing the settled sludge was filled up with mineral salts medium and was maintained in an aerobic condition by aeration for 2-4 hours. Thereafter the sludge was homogenized with a blender. The supernatant was decanted and maintained in an aerobic condition by aeration with C02-free air for 5 days. 10 mL/L were used to initiate inoculation.

- Concentration of sludge: 10 mL/L
- Initial cell/biomass concentration: 10^7 - 10^8 CFU/L
- Water: demineralized
Duration of test (contact time):
28 d
Initial test substance concentrationopen allclose all
Initial conc.:
20 mg/L
Based on:
test mat.
Initial conc.:
57.2 mg/L
Based on:
ThOD
Parameter followed for biodegradation estimation
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
Preparation of Test Flasks:
All test solutions were prepared in measuring flasks in demineralised water with the required amounts of mineral medium stock solutions and 10 mL/L inoculum. The test item was weighed out and demineralised water was added. After ultrasonic treatment it was transferred into the measuring flasks.

The reference item was weighed out and transferred into the measuring flasks with demineralised water.

250 mL measuring flasks were used to place 250 mL of the test solutions in the brown glass bottles.

A rubber sleeve with soda lime was hung into the opening of the bottles to absorb evolved CO2. The bottles were closed with Oxi-Top® measuring heads and the measuring system was activated.


Measurements:
The temperature in the incubator was documented continuously throughout the test by a hygrothermograph.
At test start the pH-values of the test solutions in the measuring flasks (before dividing) and at test end the pH-values of the test solutions in the brown glass bottles were determined.
The oxygen consumption was determined continuously from the pressure in the incubation vessels by OxiTop® measuring system at 360 measuring points (every 112 minutes) during the 28 d incubation period.



Reference substance
Reference substance:
benzoic acid, sodium salt

Results and discussion

Test performance:
Results of the Functional Control
The pass level of a biodegradation > 60 % was reached after 6 days. The validity criterion that the degradation should be > 60 % after 14 d was fulfilled

Results of the Toxicity Control
In the toxicity control the biodegradation achieved 46 % after 14 days. After 28 days the biodegradation came to 54 %. The validity criterion that the test item should not inhibit the biodegradation of the reference item was fulfilled.
% Degradation
Key result
Parameter:
% degradation (O2 consumption)
Value:
> 0 - < 10
Sampling time:
28 d
Details on results:
no biodegradation was observed

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Interpretation of results:
under test conditions no biodegradation observed
Conclusions:
The test item is classified as not readily biodegradable in the 10-d-window and after 28 days.
Executive summary:

The ready biodegradability of the test item was determined with non-adapted activated sludge in the Manometrie Respirometry Test for a period of 28 days. The study was conducted according to OECD Guideline 301 F. The test item concentration selected as appropriate was 20 mg/L, corresponding to a ThOD of 57.2 mg O2/L per test vessel. The oxygen was depleted by the respiration of bacteria and the degradation was followed by measuring the oxygen concentration. The biodegradation rate is therefore expressed as the percentage BOD (biological oxygen demand) and was calculated for each study day.

The mean oxygen depletion in the inoculum control was 7 .1 mg O2/L on day 28. In order to check the activity of the test system aniline was used as functional control. The pass level > 60 % was reached after 6 days. The biodegradation rate reached a maximum of 89 % degradation on day 24. In the toxicity control containing both test and reference item 46 % degradation occurred within 14 days. After 28 days the biodegradation came to 54 %. Both test item replicates did not reach the 10% level (beginning of biodegradation); no biodegradation was determined. The validity criteria of the Guideline are fulfilled.

After a test period of 28 days, the test item is classified as not readily biodegradable within the 10-d-window.