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Environmental fate & pathways

Biodegradation in water: screening tests

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Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 3 November 1999 to 16 August 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: EPA-Test Guideline CG-2050
Deviations:
yes
Remarks:
solutions 4 and 5
GLP compliance:
yes (incl. QA statement)
Oxygen conditions:
anaerobic
Inoculum or test system:
other: sewage, domestic
Details on inoculum:
- Laboratory culture: treatment plant at Taunusstein-Bleidenstadt
- Method of cultivation: the sludge was obtained directly from the digestor
Duration of test (contact time):
56 d
Initial conc.:
ca. 17.54 g/L
Based on:
test mat.
Details on study design:
TEST CONDITIONS
- Test temperature: 35±1ºC
- pH: 8.15

Solution 1*
0.5 g Resazurin

Solution 2*
20g (NH4)2HPO4
100 g NH4Cl

Solution 3*
23.84g CaCl2x2H2O
180g MgCl2x6H2O
130g KCl
2g MnCl2x4H2O
3g CoCl2x6H2O
0.6gH3BO3
0.23g CuCl2x2H2O
1.0g Na2MoO4x2H2O
0.2g ZnCl2

*were dissolved in 1L Ultrapure water

Solution 4+
The "solution 4" as described in the protocol (368g FeCl2x4H2O in 1L ultrapure water) was replaced by the addition of 2944 mg FeCl2x4H2O to 8000mL of final volume of the test solution.

Solution 5+
The "solution 5" as described in the protocol (50g Na2Sx9H2O in 1L ultrapure water) was replaced by the addition of 4mg Na2Sx9H2O to 8000mL of final volume of the test solution.

+This is a desviation of the protocol. The reason for this deviation is the aim of minimizing the introduction of any oxygen into the test solution.

Parameter:
other: tranformed into biogas
Value:
81
Sampling time:
56 d

Time (d) %Degradation
1 1,9
4 1,9
7 34,3
10 44
14 52
18 56
21 61
39 75
44 80
50 80
56 81
Validity criteria fulfilled:
yes
Interpretation of results:
other: biodegradable under anaerobic conditions
Conclusions:
The test substance was transformed 81% into biogas in 56 days and therefore can be considered as biodegradable under anaerobic conditions.
Executive summary:

The substance was investigated for biodegradability under anaerobic conditions according to EPA Guideline CG-2050. In the same test, inhibitory/retardant properties towards methanogenic microflora could be determined. Under the conditions of this test the test substance was transformed 81% into biogas. The control substance ethanol was degraded 95% in the same time. There was a ratardant property of the test substance which was >20% only at the beginning of the test. The mathematical value of 7.6% "inhibition" is negligible.

Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11/2/2003-12/3/2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.4-C (Determination of the "Ready" Biodegradability - Carbon Dioxide Evolution Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Oxygen conditions:
aerobic
Inoculum or test system:
other: activated sewage sludge micro-organisms
Details on inoculum:
- Source of inoculum/activated sludge: activated sewage sludge micro-organism
- Laboratory culture: aerobic stage of the Severn Trent Water Plc sewage treatment plant at Loughborough, Leicestershire, UK, which treats predominantly domestic sewage.
Duration of test (contact time):
28 d
Initial conc.:
30 mg/L
Based on:
other: suspended solid
Initial conc.:
10 mg/L
Based on:
other: carbon
Details on study design:
The following test preparation were prepared and inoculated in 5 litre glass culture vessels each containing 3 litres of solution:
a) A control, in duplicate, consisting of inoculated culture medium.
b) The standard material (sodium benzoate), in duplicate, in inoculated culture medium to give a final concentration of 10 mg carbon/L.
c) The test material, in duplicate, in inoculated culture medium to give a final concentration of 10 mg carbon/L.
d) The test material plus the standard material in inoculated culture medium to give a final concentration of 20 mg carbon/L to act as a toxicity control (one vessel only).
Reference substance:
benzoic acid, sodium salt
Parameter:
% degradation (CO2 evolution)
Value:
96
Sampling time:
28 d
Validity criteria fulfilled:
yes
Remarks:
The result og the degradtion test are considered valid if in the same test the standard material yields >= 60% degradation by Day 14.
Interpretation of results:
readily biodegradable
Conclusions:
The test material attained 96% degradation after 28 days and satisfied the 10-Day window validation criterion, whereby 60% degradation must be attained within 10 days of the degradation rate exceeding 10%. The test material can therefore be considered to be readily biodegradable under the strict terms and conditions of OECD Guideline No 301B.
Executive summary:

A study was performed to assess the ready biodegradability of the test material in an aerobic aqueous medium. The method followed that described in the OECD Guideline for Testing Chemicals (1992) No 301B, "Ready Biodegradability; CO2 Evolution Test" referenced as Method C.4 -C of Commission Directive and US EPA Fate, Tranport, and Transformation Test Guidelines OPPTS 835.3110 Paragraph (m). The test material, at a concentration of 10 mg C/L, was exposed to activated sewage sludge micro-organisms with culture medium in sealed culture vessels in the dark at 21ºC for 28 days. The degradation of the test material was assessed by the determination of carbon dioxide produced. Control solutions with inoculum and the standard material, sodium benzoate, together with a toxicity control were used for validation purposes. The test material attained 96% degradation after 28 days and satisfied the 10 -Day window validation criterion, whereby 60% degradation must be attained within 10 days of the degradation rate exceeding 10%. The test material can therefore be considered to be readily biodegradable under strict terms and conditions of OECD Guideline No 301B.

Description of key information

Aerobic biodegradation: The test material attained 96% degradation after 28 days and satisfied the 10-Day window  validation criterion, whereby 60% degradation must be attained within 10 days of the degradation rate exceeding 10%. 
Anaerobic biodegradation: In a test according to EPA Guideline CG-2050 the test substance was transformed 81% into biogas.

Key value for chemical safety assessment

Biodegradation in water:
readily biodegradable

Additional information