An experimental procedure was developed for the respirometric determination of bacterial storage yield as defined in the Activated Sludge Model No. 3. The proposed approach is based on the oxygen utilization rate (OUR) profile obtained from a batch test and correlates the area under the OUR curve to the amount of oxygen associated with substrate storage. Model simulation was used to evaluate the procedure for different initial experimental conditions. The procedure was tested on acetate. The same storage yield value of 0.76 gCOD/gCOD was calculated for two experiments, starting with different F/M ratios of 0.12 and 0.64 gCOD/g cellCOD.