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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:
From April 04 to July 22, 2019
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report

Materials and methods

Test guidelineopen allclose all
according to guideline
OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
according to guideline
EPA OPPTS 835.3110 (Ready Biodegradability)
Principles of method if other than guideline:
Test duration: 28 days.
Test environment: Thermostat.
Monitoring of test medium pH: At the start and at the end of the test, in all test item flasks.
Test temperature: 21.8 – 22.1 °C.
pH of test medium: 7.12– 7.43 at the test start.
7.48 – 8.27 at the test end.
GLP compliance:
yes (incl. QA statement)

Test material

Chemical structure
Reference substance name:
Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized
EC Number:
EC Name:
Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized
Cas Number:
Molecular formula:
Not applicable - UVCB substance
Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized
Test material form:
Specific details on test material used for the study:
Test item name: Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized
Batch number: I1-0000196125
Manufacturing date: July 28, 2018
Expiry date : July 28, 2019
C.A.S. number: 90459-71-5
EEC number : 291-716-4
Molecular weight: UVCB substance
Storage : The test item will be stored at room temperature, in the dark

Study design

Oxygen conditions:
Inoculum or test system:
activated sludge, domestic (adaptation not specified)
Details on inoculum:
The study was performed using aerobic activated sludge from BRIANZACQUE civil wastewater treatment plant. A homogenized aliquot of the wet sludge was centrifuged, the supernatant was removed while the solid phase was weighed, dried and the ratio of wet to dry weight was calculated to be equal to 5.37%.
Based on this ratio, calculated amounts of wet sludge was centrifuged, the supernatant was removed and the solid phase was suspended in the test medium to get a concentration equivalent to 3 g dry material per litre.
The prepared sludge inoculum was pre-conditioned to the experimental conditions by aerating it in the dark. The concentrated suspension was used as inoculum to give a final concentration of 15 mg dry material per litre into test flasks.
Duration of test (contact time):
28 d
Initial test substance concentrationopen allclose all
Initial conc.:
50 mg/L
Based on:
test mat.
Initial conc.:
67 mg/L
Based on:
Details on study design:
Since the test item was soluble in water, a concentrated stock solution at nominal concentration of 200 mg/L was prepared by weighing 0.0505 g of test item in 250 mL of mineral medium (corresponding to 202 mg/L corrected for the weighed test item).
The proper volume was dosed in the test flasks to obtain the final concentrations of 50 mg/L of test item making to a final volume of 250 mL or 150 mL (based on setting scale) with test medium.

Moreover, stock solutions (10 g/L) of reference item Sodium Benzoate and sterilizing agent HgCl2 were prepared and an appropriate aliquot of the stock solution was added to the test medium (final volume: 250 mL or 150 mL ) in the test flasks to obtain a final concentration of 100 mg/L.

Activated sludge was added to each flask (with the exception of the abiotic control) before incubation.
Finally, the test flasks, containing stir bars covered with teflon, were kept under continuous stirring by the means of a magnetic stirrer for the 28-days test period.

The incubation temperature was continuously monitored during the course of the study by means of a data logger. It was in the range 21.8 – 22.1 °C with a mean value of 22.0 °C and a standard deviation of 0.04 °C. The temperature range recommended by OECD guideline is 22 ± 2 °C, this range was corrected according to the data logger precision, therefore the range to be considered for the test was 20.4 – 23.6 °C while the recorded temperature values were corrected according to the correction factor.
The recorded temperature values during the test period were within the recommended range
and the maximum deviation was lower than 1 °C, as recommended by the guideline.

Reference substance
Reference substance:
benzoic acid, sodium salt

Results and discussion

% Degradation
% degradation (O2 consumption)
Sampling time:
28 d
Details on results:
Test item concentration should be 100 mg/L or a concentration giving at least 50-100 ThOD or COD/L.
The tested concentration was 50 mg/L (corresponding to 67 mg COD/L).
All reported results are related to the nominal concentration of test item.

The biodegradation was followed by measuring the oxygen uptake (mg/L) of the test item under defined conditions by the means of a manometric method and calculating from these values the BOD (Biochemical Oxygen Demand).

BOD5 / COD results

1.34 g O2/g test mat.
Results with reference substance:
COD= 1.67 mg O2/mg for reference item
% degradation= 79.6%

Applicant's summary and conclusion

Validity criteria fulfilled:
Interpretation of results:
not readily biodegradable
The 10-day window wasn’t start during test period.
Maximum value at the end of the test period (28 days of exposure): 5.2 % (mean percentage values between two test item replicates).

The test item, Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized, was found to be not ready and completely biodegradable under the conditions applied in a manometric respirometry test, having showed a maximum biodegradation of 5.2% at the end of the test (mean percentage values between two replicates),and the 10-day window wasn’t start.

No significant degradation was observed in the abiotic control.

The test item showed no inhibitory effects on the microorganism activity at the tested concentration, as demonstrated by the biodegradation percentage in the toxicity control exceeding the 25% pass level within 14 days of exposure (29.1%).

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