Optimisation of ATP determination in drinking water: Manuscript

Adenosine Triphosphate (ATP) can be used as a relative measure of cell activity, and is measured by the light output from the reaction between luciferin and ATP catalyzed by firefly luciferase. The measurement has potential as a monitoring and surveillance tool within drinking water distribution, since the method is very sensitive (detects 0.5 ng ATP/L) and results are obtained within minutes. When calculating the ATP value a number of parameters need to be considered. These were investigate by use of two different reagent kits (PCP-kit and Lumin(ATE)/Lumin(EX)-kit), internal standard and an Advance Coupe luminometer. The
investigations showed a 60 times higher response of the PCP-kit, making it more suitable for measurement of samples with low ATP content. ATP-standard dilutions prepared in tap water were stable for at least 15 months when stored frozen at -80ºC, and storage of large aliquots of standards increase quality control and ease daily operation. The medium (Lumin(PM) buffer, tap water or MilliQ water) for preparation of ATP-standard dilution significantly affected the rlu
response of the ATP-standard dilutions (20% difference). The effect of dilution media and of sample characteristics can be eliminated by use of internal standard. In strongly coloured biofilm samples the measuring efficiency can be reduced with up to 85%. Extra cellular ATP made up a significant part of the total ATP (>50%) in some samples, so when only intra cellular ATP is of interest the cells need be separated from the water phase by filtration.