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A measured volume of inoculated mineral medium, containing the test item as the nominal sole source of organic carbon is aerated by the passage of carbon-dioxide free air at a controlled rate in the dark. Degradation is followed over 28 days by determining the carbon dioxide produced.
The CO2 is trapped in barium hydroxide and is measured by titration of the residual hydroxide.
The amount of carbon dioxide produced from the test item (corrected for that derived from the blank inoculum)is expressed as a percentage of ThCO2 (theoretical CO2).
T1a – T1b = containing test substance and inoculum (test suspension)
Ctrla – Ctrlb = containing only inoculum (blank)
Ra = containing reference item and inoculum
and when necessary
Aa = containing test substance and sterilising agent (abiotic sterile control)
Xa = containing test substance, reference compound and inoculum (toxicity control)
Abiotic sterile control (A) is used to check if the test substance is degraded abiotically by using a sterilised uninoculated solution of the chemical.
Toxicity control (X) is used to check the possible inhibitory effect of the test substance by adding it at the same concentration as present in the other flasks.
% of Degradation after 28 days
REFERENCES AND GUIDELINES
OECD Guideline for Testing of Chemicals, No. 301-B (17thJuly 1992)-CO2 Evolution Test