Analyzing adenosine nucleotides, such as ATP, ADP, and ADP, was found to be difficult using standard LC systems and columns made of stainless-steel components. The challenge can be attributed to the strong affinity of these compounds towards exposed metal surfaces. The analytes with two or three phosphate groups showed the worst effects. With the standard LC and column, ATP and ADP were completely lost at and below 2 ng/µL sample concentrations. AMP, with a single phosphate group, showed comparatively minor losses. Nonetheless, it suffered from compromised assay sensitivity.
As shown with this case study, the ACQUITY PREMIER System can be effectively used with an ACQUITY PREMIER Column to overcome some metal adsorption challenges that can interfere with achieving robust quantitation. With this technology, new levels of performance will now become possible for the quantitation of nucleotides, whether the employed technique be based on a fit-for-purpose RPLC method, HILIC, or mixed mode chromatography.