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Environmental fate & pathways

Biodegradation in water: screening tests

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Endpoint:
biodegradation in water: inherent biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
9 December 1996 - 7 January 1997
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
GLP compliance is not specified
Qualifier:
according to guideline
Guideline:
OECD Guideline 302 B (Inherent biodegradability: Zahn-Wellens/EMPA Test)
Deviations:
no
GLP compliance:
not specified
Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, industrial (adaptation not specified)
Details on inoculum:
Inoculum was prepared from activated sludge taken from Perstorp AB waste water treatment plant, and was washed twice with mineral medium prior to sampling. The activated sludge contained 15.4 g suspended solids per litre.
Duration of test (contact time):
28 d
Initial conc.:
0.5 other: ml/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
DOC removal
Details on study design:
The following test, control, inhibition, sterile and blank mediums were derived for use in the study.

Test medium
The following solutions were added to derive 1 L test medium:
- 10 ml solution A
- 1 ml (each) of solutions B to D
- 0.5 ml test solution (Pentaerythritol triallyl ester)
- Approximately 900 ml deionised water. Solution pH was adjusted to 7.4
- 65 ml inoculum
Medium was topped up to 1000 ml with deionised water.

Control medium
The following solutions were added to derive 1 L control medium:
- 10 ml solution A
- 1 ml (each) of solutions B to D
- 10 ml stock solution 1
- Approximately 900 ml deionised water. Solution pH was adjusted to 7.4
- 65 ml inoculum
Medium was topped up to 1000 ml with deionised water.

Inhibition medium
The following solutions were added to derive 1 L inhibition medium:
- 10 ml solution A
- 1 ml (each) of solutions B to D
- 0.5 ml test solution (Pentaerythritol triallyl ester)
- 10 ml stock solution 1
- Approximately 900 ml deionised water. Solution pH was adjusted to 7.4
- 65 ml inoculum
Medium was topped up to 1000 ml with deionised water.

Sterile medium
The following solutions were added to derive 1 L sterile medium:
- 10 ml solution A
- 1 ml (each) of solutions B to D
- 0.5 ml test solution (Pentaerythritol triallyl ester)
- 10 ml stock solution 2
- Approximately 900 ml deionised water. Solution pH was adjusted to 7.4
Medium was topped up to 1000 ml with deionised water.

Blank medium
The following solutions were added to derive 1 L blank medium:
- 10 ml solution A
- 1 ml (each) of solutions B to D
- Approximately 900 ml deionised water. Solution pH was adjusted to 7.4
- 65 ml inoculum
Medium was topped up to 1000 ml with deionised water.


Test:

4 L of test medium was prepared and 2 L added in duplicate to 2 L flasks.
2 L of control medium was prepared and added to a 2 L flask.
2 L of inhibition medium was prepared and added to a 2 L flask.
0.5 L of sterile medium was prepared and added to a 0.5 L flask.
2 L of blank medium was prepared and added to a 2 L flask.

Each flask was assigned the project number, date and type of medium, and the level of medium was marked on the flask.
Flasks were all incubated in a room with diffused light at room temperature (22-24°C). To maintain aerobic conditions in mediums and obtain stirring, flasks were aerated through long glass tubes which ended at the bottom of the flasks.

Prior to sampling, water losses due to evaporation were made up with deionised water. After sampling, a new mark was made at the new medium level.
Dissolved organic carbon (DOC) concentrations were measured for each period (0, 3h, 3, 5, 7, 14, 21 and 28 days) in each inoculated flask. The total organic carbon (TOC) concentration was measured at day 0 and 28. The DOC concentration in the sterile medium flask was only measured on day 0 and 28. A Scheicher & Schuell filter (0.45 µm, OE 67, 45 mm) was used for DOC samples, and boiled 3 times in deionised water to purify the filters from soluble carbon. Each boiling period lasted for at least one hour.
Reference substance:
ethylene glycol
Parameter:
% degradation (DOC removal)
Value:
11
Sampling time:
29 d
Remarks on result:
other: Test medium D(TI)
Parameter:
% degradation (DOC removal)
Value:
11
Sampling time:
29 d
Remarks on result:
other: Test medium D(TII)
Details on results:
The quantity DOC that is degraded in the toxicity control flask (FI) corresponds to the part that comes from the reference compound, thus showing that the test article was no inhibitory to the inoculum. The percentage DOC reduction measured after 28 days (at day 29) in both test mediums TI and TII was 11% after correction for the DOC contribution from the inoculum.
Results with reference substance:
The degradation of the reference compound (ethylene glycol) was approximately 100% (i.e. >70%) after days, thus the study is considered to be valid.

TOC-content (mg/L) during the study

Day F(B) F(C) F(TI) F(TII) F(I) F(S) D(TI) D(TII)
0 25 430 330 320 720 270 0 0
29 16 38 280 290 290 240 13

7

DOC-content (mg/L) during the study

Day F(B) F(C) F(TI) F(TII) F(I) F(S) D(TI) D(TII) D(C)
0 5 430 290 290 740 280 0 0 0
0.125 5.8 400 270 270 730 - 7 7 7
3 5.7 180 270 290 480 - 7 0 59
5 6.1 20 290 280 320 - 0 4 97
7 7 10 280 280 290 - 1 1 99
14 6.3 8 280 260 270 - 4 11 100
21 6.3 7 270 260 280 - 7 11 100
29 6.8 8 260 260 280 260 11 11 100

FT= Test flasks containing sample and inoculum

FS= Control flask for non-biological degradation containing sample and mercury chloride

FB= Control flask for DOC contribution from the inoculum

FC= Flask containing the reference compound (ethylene glycol) for checking inoculum activity

FI= Flask containing sample and the reference compound (ethylene glycol) to check for possible inhibitory effect of the sample on the inoculum

DX= percent DOC reduction compared to day 0 after correction for the DOC contribution from the inoculum

Validity criteria fulfilled:
yes
Remarks:
Reference compound degradation was approximately 100% after 14 days
Interpretation of results:
not inherently biodegradable
Conclusions:
Dissolved organic carbon (DOC) removal for the test solution does not exceed 70% after 28 days, therefore it cannot be regarded as ultimately biodegradable.
Executive summary:

The percentage DOC reduction measured after 28 days (at day 29) in both test mediums TI and TII was 11% after correction for the DOC contribution from the inoculum.  1,3-propanediol, 2,2- bis(hydroxymethyl) -, allyl ether therefore cannot be regarded as ultimately biodegradable.

Endpoint:
biodegradation in water: ready biodegradability
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:

Description of key information

An OECD 302B inherent biodegradability study is available for 1,3-propanediol, 2,2- bis(hydroxymethyl) -, allyl ether, the results of which showed that the percentage dissolved organic carbon (DOC) reduction measured after 28 days (at day 29) in both test mediums (TI and TII) was 11% after correction for the DOC contribution from the inoculum.  1,3-propanediol, 2,2- bis(hydroxymethyl) -, allyl ether therefore cannot be regarded as ultimately biodegradable.

Key value for chemical safety assessment

Biodegradation in water:
under test conditions no biodegradation observed

Additional information