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Du Pont Instruments

In a detailed study of the control parameters for the activated sludge process, the measurements of interest are BOD, COD, BOD and COD reduction, biological population density, and biological oxidative activity. The amount of ATP is proportionate to the viable biomass in a sample, whereas changes in ATP concentration measure the biomass oxidative capability. Thus, environmental engineers are interested in measuring the ATP content of samples in the activated sludge process as well as in rivers, lakes, and other receiving waters.

An ATP assay procedure has been developed based on the reactions just described. Briefly, the procedure involves the rapid killing of live bacterial cells and the immediate extraction of ATP into an aqueous solution. The latter is then treated with firefly lantern extract, and a photometer measures the light emission of the resultant solution. The firefly lantern extract and the ATP required for calibration are commercially available. Du Pont is the only supplier who has designed a manually operated instrument specifically for this measurement.

The instrument is supplied with the required reagents. The lab technician dissolves a tablet containing buffer and magnesium sulfate in water and adds a homogeneous powder of luciferin and luciferase. Then, the technician filters the sample through a coarse filter to remove solid matter, which is discarded. The filtrate passes through a bacterial filter to catch the living bacteria. The technician treats the bacteria on the filter with butanol, which ruptures the cell walls and releases the ATP. To take the filtrate up to volume, the technician adds water and a microliter aliquot to the prepared reagent already in a cuvette. He then places the cuvette in the instrument to read its light emission. The instrument automatically converts the light flash to ATP or microorganism concentration per milliliter, depending on how it is calibrated.

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