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Environmental fate & pathways

Biodegradation in water: screening tests

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Reference
Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24.07.1997 - 14.04.1998
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
Version / remarks:
Adopted by the Council on 17th July 1992
Qualifier:
according to guideline
Guideline:
EU Method C.4-D (Determination of the "Ready" Biodegradability - Manometric Respirometry Test)
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
stable in water
Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, domestic, non-adapted
Details on inoculum:
Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure):

- Species/Origin: Aerobic activated sludge, microorganisms from a domestic waste water treatment plant (Sindlingen, Frankfurt am Main, Germany).
- Concentration of sludge: stock suspension of 2.2 g/L on dry matter base - final sludge concentration in test flasks: 22 mg sludge/L
Duration of test (contact time):
28 d
Initial conc.:
96.24 mg/L
Based on:
COD
Initial conc.:
34.25 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
TEST CONDITIONS
- Composition of medium: Mineral medium was prepared from stock solutions of appropriate concentrations of mineral components, as described in the OECD Guideline.
- Test temperature: 22°C +- 1°C

TEST SYSTEM
- Number of flasks/concentration: Two test flasks were used with 34.25 mg/L and 34.25 mg/L per flask.

CONTROL AND BLANK SYSTEM
- Inoculum blank: Two flasks were prepared with activated sludge and mineral medium.
- Toxicity control: Two flasks with 35.25 mg/L and 34.50 mg/L test item and 60 mg/L sodium benzoate per flask.
- Other: One flask for the procedure control with 60 mg/L mg sodium benzoate.
Reference substance:
benzoic acid, sodium salt
Parameter:
% degradation (O2 consumption)
Value:
4
Sampling time:
5 d
Remarks on result:
other: Data are averages of two independent tests
Parameter:
% degradation (O2 consumption)
Value:
43
Sampling time:
11 d
Remarks on result:
other: Data are averages of two independent tests
Parameter:
% degradation (O2 consumption)
Value:
61
Sampling time:
17 d
Remarks on result:
other: Data are averages of two independent tests
Parameter:
% degradation (O2 consumption)
Value:
66
Sampling time:
28 d
Remarks on result:
other: Data are averages of two independent tests
Details on results:
The criterion for ready biodegradability under the conditions of a manometric respirometry test is the degradation of the test item of at least 60%, reached within a 10-day window; the 10-day window starts when the degradation of the test item reaches at least 10% degradation.

The test system showed no inhibitory/toxic effects at the concentration applied. The pass level of 60 % COD was not completely reached within the 10-d window. As the compound is a mixture, it could not be excluded that individual components are not or only partially biodegraded.

--------------

Kinetic of control substance (in %):
ca. 65 after 3 day(s)
ca. 93 after 14 day(s)
Degradation products: not measured
Parameter:
COD
Value:
2 810 mg O2/g test mat.
Results with reference substance:
The reference item sodium benzoate was sufficiently degraded to 65% after 3 days and to 93% after 14 days of incubation, thus confirming the suitability of the aerobic activated sludge inoculum used.

Oxygen consumption in mg/L and degradation in % for test item:

    Oxygen consumption in mg/L minus mean values of blank control        Degradation in %
 Time  Flask A  Flask C  Flask A Flask C  Mean value 
 1 d  0  0
 3 d  0 0  0
 5 d  6.3  7
 8 d  43.30  0  45 23 
 11 d  54.00  28.60  56 30  43 
 14 d  57.40  51.10  60 53  57 
 17 d  59.45  56.85  62 59  61 
 21 d  60.75  60.95  63 63  63 
 24 d  61.70  63.20  64 66  65 
 28 d  62.50  64.80  65 67  66 
Validity criteria fulfilled:
yes
Interpretation of results:
inherently biodegradable
Conclusions:
The criterion for ready biodegradability under the conditions of a manometric respirometry test is the degradation of the test item of at least 60%, reached within a 10-day window; the 10-day window starts when the degradation of the test item reaches at least 10% degradation.

The mean biodegradation at test end after 28 days was 66% (COD). However, the pass level of 60 % COD was not completely reached within the 10-d window, since the adaptation period was 6 days (biodegradation above 10%) and the pass level was reached after 17 days based on the man values. As the compound is a mixture, it could not be excluded that individual components are not or only partially biodegraded.

Therefore, the test substance is considered to be inherently biodegradable.
Executive summary:

The test item (initial concentration: 34.25 mg/L, two replicates) was investigated compliant with GLP for its ready biodegradability in a manometric respirometry test according to OECD 301F over a period of 28 days. The biodegradation was followed by the oxygen uptake of the microorganisms during exposure. As a reference item sodium benzoate was tested simultaneously under identical conditions (procedural control). Replicate inoculum controls and toxicity controls (two flasks) containing test item and reference item at the same concentrations as provided in test and reference item flasks were set up and run in parallel. The test met all validity criteria as given in OECD guideline 301.

Under the test conditions, C13/C15 amine reached 66% biodegradation at the end of the test (28 days, mean value of both replicates). Biodegradation of the reference item demonstrated proper functioning of the test system (biodegradation extent 93% after 14 days). Since the biodegradation of at least 60% was not reached within a 10 -day window, the test item is considered to be inherently biodegradable under the conditions of this test.

Based on the results of the toxicity control containing test item and reference item at the same concentrations as provided in the test and reference flasks, the test item was not inhibitory with regard to microorganisms responsible for biodegradation of the reference compound.

Description of key information

Inherently biodegradable (OECD 301 F; GLP): 66 % within 28 days based on biological oxygen demand of two replicates (mean value).

The pass level for ready biodegradability was not reached in a 10-d window, since it started at day 6 (degree of biodegradation has reached at least 10%) and 66% removal of DOC were reached after 28 days (mean value of two replicates).

Key value for chemical safety assessment

Biodegradation in water:
inherently biodegradable
Type of water:
freshwater

Additional information

The test item (initial concentration: 34.25 mg/L, two replicates) was investigated compliant with GLP for its ready biodegradability in a manometric respirometry test according to OECD 301F over a period of 28 days. The biodegradation was followed by the oxygen uptake of the microorganisms during exposure. As a reference item sodium benzoate was tested simultaneously under identical conditions (procedural control). Replicate inoculum controls and toxicity controls (two flasks) containing test item and reference item at the same concentrations as provided in test and reference item flasks were set up and run in parallel. The test met all validity criteria as given in OECD guideline 301.

Under the test conditions, C13/C15 amine reached 66% biodegradation at the end of the test (28 days, mean value of both replicates). Biodegradation of the reference item demonstrated proper functioning of the test system (biodegradation extent 93% after 14 days). Since the biodegradation of at least 60% was not reached within a 10 -day window,the test item is considered to be inherently biodegradable under the conditions of this test.

Based on the results of the toxicity control containing test item and reference item at the same concentrations as provided in the test and reference flasks, the test item was not inhibitory with regard to microorganisms responsible for biodegradation of the reference compound.