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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:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
Deviations:
yes
Remarks:
Determination of the Chemical Oxygen Demand (COD), was not performed under GLP
Qualifier:
according to guideline
Guideline:
EU Method C.4-D (Determination of the "Ready" Biodegradability - Manometric Respirometry Test)
Deviations:
yes
Remarks:
Determination of the Chemical Oxygen Demand (COD), was not performed under GLP
Qualifier:
according to guideline
Guideline:
EPA OPPTS 835.3110 (Ready Biodegradability)
Deviations:
yes
Remarks:
Determination of the Chemical Oxygen Demand (COD), was not performed under GLP
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Identification: FAT #: 93580/A
Batch: 0022357000
Purity: 94.3 %
Expiry date: 15 December 2015
Description at Room Temperature: White-yellowish waxy solid
Storage Conditions: Room temperature at 20 ± 5 °C, in the dark.
Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, domestic (adaptation not specified)
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): wastewater treatment plant (ARA Ergolz II, Füllinsdorf, Switzerland)
- Preparation of inoculum for exposure: The sludge was washed twice with tap water by centrifugation and the supernatant liquid phase was decanted. A homogenized aliquot of the final sludge suspension was weighed, thereafter dried and the ratio of wet to dry weight was calculated.
- Pretreatment: During the holding period of one day prior to use, the sludge was aerated at room temperature. Prior to use, the sludge was first thoroughly mixed and then diluted with test water to a concentration of 1 g per liter (dry weight basis). Based on the determined dry weight of this diluted activated sludge, defined amounts were added to test water to obtain a final concentration of 30 mg dry material per liter.
- Concentration of sludge:30 mg dry material per liter.
Duration of test (contact time):
28 d
Initial conc.:
100 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
TEST CONDITIONS
- Composition of medium: Test water along with 30mg (dry material per liter) of activated sludge
- Solubilising agent (type and concentration if used): None
- Test temperature: 22 °C, maintained with a built-in thermostat and checked at least at each working day.
- pH: 7.4-7.6
- pH adjusted: no
- Continuous darkness: yes


TEST SYSTEM
- Culturing apparatus: The test flasks (500 mL reaction vessels, labelled with the necessary information to ensure unmistakable identification) were incubated under continuous stirring in a SAPROMAT D12 (Voith GmbH, Heidenheim, Germany).
- Number of culture flasks/concentration: Duplicate
- Measuring equipment:
Electro-chemical analysis process:
The biodegradation process consumes the dissolved oxygen in the test medium and generates CO2. The CO2 is adsorbed by soda lime, which results in a decrease of the total pressure in the airtight test flasks. The pressure drop is detected and converted into an electrical signal by means of an electrode type manometer. The consumed oxygen is replaced by electrolytically generated oxygen from a copper sulfate solution.

SAMPLING:
The test flasks (500 mL reaction vessels, labelled with the necessary information to ensure unmistakable identification) were incubated under continuous stirring in a SAPROMAT D12 (Voith GmbH, Heidenheim, Germany). Oxygen consumption was recorded manually by taking a daily reading at least on each working day.

CONTROL AND BLANK SYSTEM
- Inoculum blank: Yes
- Toxicity control: Yes

STATISTICAL METHODS: None employed
Reference substance:
benzoic acid, sodium salt
Preliminary study:
The percent biodegradation of the test item was calculated based on the chemical oxygen demand (COD) of 2.47 mg O2/mg test item.
The biochemical oxygen demand (BOD) of FAT 93580/A in the test media was in the range of the inoculum controls throughout the study period of 28 days.
At Exposure Day 14 the mean biodegradation of FAT #: 93580/A reached 3 % and remained there until the end of the 28-day exposure period.
Test performance:
The results are considered valid since the following criteria are met:
- The percentage degradation of the reference item reached the level for ready biodegradability by Day 3 (criterion: at least 60 % of ThOD in a 10-day window by Day 14 for the reference item).
- The mean oxygen demand of the inoculum control (medium and inoculum) was 18 mg O2/L within 28 days (criterion: normally 20-30 mg O2/L, but not greater than 60 mg O2/L). Since the test item was not degraded, other validity criteria are not applicable.
Key result
Parameter:
% degradation (O2 consumption)
Value:
3
Sampling time:
28 d
Details on results:
The percent biodegradation of the test item was calculated based on the chemical oxygen demand (COD) of 2.47 mg O2/mg test item.
The biochemical oxygen demand (BOD) of FAT #: 93580/A in the test media was in the range of the inoculum controls throughout the study period of 28 days.
At Exposure Day 14 the mean biodegradation of FAT #: 93580/A reached 3 % and remained there until the end of the 28-day exposure period.
Consequently, FAT #: 93580/A was not biodegradable under the test conditions within 28 days.

The results are considered valid since the following criteria are met:
- The percentage degradation of the reference item reached the level for ready biodegradability by Day 3 (criterion: at least 60 % of ThOD in a 10-day window by Day 14 for the reference item).
- The mean oxygen demand of the inoculum control (medium and inoculum) was 18 mg O2/L within 28 days (criterion: normally 20-30 mg O2/L, but not greater than 60 mg O2/L).
Results with reference substance:
The percent biodegradation of the reference item sodium benzoate was calculated based on the theoretical oxygen demand (ThOD) of 1.67 mg O2/mg reference item.
In the procedure controls, the reference item was degraded by an average of 87% by Exposure Day 14, thus confirming suitability of the activated sludge. At Day 28, the reference item was degraded by an average of 98%.

Biodegradation in the Toxicity Control:

In the toxicity control, containing both the test item and the reference item sodium benzoate, the biodegradation amounted to 36% within 14 days, reaching 39% at the end of the test. Therefore, according to the test guidelines, FAT #: 93580/A had no inhibitory effect on activated sludge microorganisms at the tested concentration of 101 mg/L because biodegradation in the toxicity control was >25 % within 14 days.

Validity criteria fulfilled:
yes
Interpretation of results:
not readily biodegradable
Conclusions:
FAT #: 93580/A was not biodegradable under the test conditions within 28 days.
Executive summary:

The test item FAT #: 93580/A was investigated for its ready biodegradability in a manometric respirometry test over 28 days according to the OECD Guideline for Testing of Chemicals (1992) No. 301F, the Commission Regulation (EC) No 440/2008, C.4-D and the US EPA Draft Ecological Effects Test Guidelines OPPTS 835.3110 (Paragraph (q)).

The percent biodegradation of the test item was calculated based on the chemical oxygen demand (COD) of 2.47 mg O2/mg test item.

The biochemical oxygen demand (BOD) of FAT #: 93580/A in the test media was in the range of the inoculum controls throughout the study period of 28 days. At Exposure Day 14 the mean biodegradation of FAT #: 93580/A reached 3 % and remained there until the end of the 28-day exposure period.

The reference item sodium benzoate was degraded by an average of 87 % by Exposure Day 14, and reached an average biodegradation of 98 % by the end of the test (Day 28), thus confirming suitability of the activated sludge.

Consequently, FAT #: 93580/A was not biodegradable under the test conditions within 28 days.

Description of key information

The test item FAT 93580/A was investigated for its ready biodegradability in a manometric respirometry test over 28 days according to the OECD Guideline for Testing of Chemicals (1992) No. 301F, the Commission Regulation (EC) No 440/2008, C.4-D and the US EPA Draft Ecological Effects Test Guidelines OPPTS 835.3110 (Paragraph (q)).

The percent biodegradation of the test item was calculated based on the chemical oxygen demand (COD) of 2.47 mg O2/mg test item. The biochemical oxygen demand (BOD) of FAT 93580/A in the test media was in the range of the inoculum controls throughout the study period of 28 days. At Exposure Day 14 the mean biodegradation of FAT 93580/A reached 3 % and remained there until the end of the 28-day exposure period.

Consequently, FAT 93580/A was not biodegradable under the test conditions within 28 days.

Key value for chemical safety assessment

Biodegradation in water:
under test conditions no biodegradation observed

Additional information