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Diss Factsheets

Environmental fate & pathways

Biodegradation in water: screening tests

Administrative data

biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Feb - Mar 2001
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
GLP compliance:

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
Cas Number:
Molecular formula:
C21 H26 O3
Test material form:
solid: bulk

Study design

Oxygen conditions:
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): Kläranlage Berlin Ruhleben
- Method of cultivation: homogenized aerated and stirred
Duration of test (contact time):
28 d
Initial test substance concentration
Initial conc.:
10 mg/L
Based on:
Parameter followed for biodegradation estimation
Parameter followed for biodegradation estimation:
CO2 evolution
Details on study design:
- Test temperature: 23 - 24 °C
- pH: 7.5 - 7.8
- Continuous darkness: yes
- pH adjusted: yes

- Number of culture flasks/concentration: 3
- Method used to create aerobic conditions: Aerated with CO2-free air
- Details of trap for CO2 and volatile organics if used: For the determination of the CO2 produced, three CO2 absorber bottles, filled with 100 mL 0.025 N Ba(OH)2 each, were connected in series to the exit air-pipe of each test bottle. The amount of CO2 produced was determined by titration of the remaining Ba(OH)2 with 0.05 N standardized HCI. On days 4, 6, 8, 11, 14, 19, 22, 2'7 and 29 the CO2 absorber bottle nearest to the test bottles was removed for the titration, with the exception of day 29 when all three bottles were removed. The remaining two absorbers were each moved one place closer to the test vessel and a new absorber bottle filled with fresh Ba(OH)2 was placed at the far end of the series. In cases where BaCO3 was also precipitated in the second absorber bottle of any solution, titrations of two Ba(OH)2 bottles were performed. Subsequently, two fresh absorbers were added.

- Sampling frequency: 4, 6,8, 11, 14, 19, 22, 27, 29 days; After the measurements of pH, 1 mL of concentrated HCl was added to each test vessel in order to convert all dissolved inorganic carbon into CO2.

- Inoculum blank: yes, 3 vessels
- Abiotic sterile control: no
- Toxicity control: yes, 1 vessel
- Reference substance: yes, 1 vessel
Reference substance
Reference substance:
benzoic acid, sodium salt

Results and discussion

% Degradation
Key result
% degradation (CO2 evolution)
Sampling time:
28 d

BOD5 / COD results

Results with reference substance:
The reference substance was completely degraded after 28 d.

Any other information on results incl. tables

Table 1: Amounts of CO2 produced (cumulative), in mg (values for ZK 5668, sodium acetate and toxicity control corrected for blank CO,: production)

 Control/blank (mean of 3 vessels)   Concentration of carbon  2,956,508,610,612,715,517,722,6
 ZK 5668 (mean of 3 vessels)   0 mg/L  2,83349,906474,682,685,388,390,2
 Reference (sodium acetate)   10 mg/L  42,565,775,0085,59399,5101103,8105,3
 Toxicity control (ZK 5668 + sodium acetate)   10 mg/L + 10 mg/L  39,892117,90141,7157,9170,3176,1181,7185,2

Applicant's summary and conclusion

Validity criteria fulfilled:
Interpretation of results:
readily biodegradable
According to the results of the test it can be concluded that the subsance is readily biodegradable under the conditions of the test.
Executive summary:

The purpose of this study was to determine the ready biodegradability of 1,4,6-Trienol (ZK 5668). The study was conducted in agreement with the OECD test guideline no. 301 B.

The test substance was incubated in an aqueous solution including nutrients with microorganisms from a municipal sewage treatment plant for 28 days (start of treatment = day 1). The nutrient solutions were made up of phosphates, magnesium sulphate, iron chloride, ammonium chloride and calcium chloide. The test substance was incubated at a concentration of 10 mg carbon/L in triplicate. Additionally, a reference substance (sodium acetate) was tested in a single set according to the same procedure, in order to verify the viability and activity of the degrading microorganisms. One further set was incubated with sodium acetate at 10 mg carbon/L (reference substance) plus test substance at 10 mg carbon/L representing a toxicity control. Furthermore, a blank control was tested in triplicate without any test or refeference substance. The biological degradation of the test and reference substances was evaluated by measurement of the carbon dioxide (CO2) produced during the test period. CO2 production was determined on days 4, 6, 8, 11, 14, 19, 22, 27 and :29. On day 28 the solutions were acidified in order to expel all dissolved CO2 and CO2 was determined on day 29. The CO2 production was calculated as the percentage of total CO2 that the testmaterial could theoretically have produced, based on carbon content. The blank CO2 production was subtracted for correction.

The test compound 1,4,6 -Trienol was degraded to 68% on day 14 and 82% on day 29 (28 days of incubation). The reference compound sodium acetate was degraded to 60% on day 6 and 96% on day 29 (28 days of incubation). In the toxicity control, the reference compound (sodium acetate) plus the test compound, was degraded to 84% on day 29 (28 days of incubation), reflecting the degradation in the individual sets.

Based on the result, the substance is considered being readily biodegradable according to the OECD criteria.