A rationale for the appropriate amount of inoculum in ready biodegradability tests

Several screening methods at the so-called ready biodegradability level are suitable to test poorly soluble substances. Typical for these tests is that mineralization is evaluated from monitoring oxygen uptake or carbon dioxide production. Unfortunately, they suffer from a rather low precision in the calculated percentage of mineralization caused by subtracting a too high inoculum control measurement from the response in the test system. Criteria for blank oxygen consumption, due to the metabolic activity of the inoculum, are proposed from which maximum amounts of activated sludge or secondary effluent per litre test medium can be derived to be used as an appropriate inoculum. Both for current and future standardized tests the precision of the method can be kept within acceptable margins. Inoculum material was sampled from 40 communal biological waste water treatment plants. From endogenous respiration rates it was derived that the concentration of secondary effluent in the Closed Bottle Test can be increased up to 50 mL/L but that in respirometry tests inoculated with activated sludge the appropriate concentration is 10 mg/L dry matter or below, depending of the design of the test system.