ATP, ADP and AMP profiling for diagnostic applications: Recent advances in analytical strategies

Adenylate compounds – adenosine triphosphate (ATP), diphosphate (ADP), and monophosphate (AMP) – are central regulators of cellular energy metabolism and biomarkers of physiological and pathological states. Their rapid interconversion and enzymatic lability make accurate quantification challenging, especially in complex matrices. This review summarizes strategies for the analysis of ATP, ADP, and AMP, focusing on sample preparation and analytical methodologies for biological systems. Analytical techniques – including bioluminescent assays, biosensors, and chromatographic methods – have improved sensitivity and throughput, but most remain limited to endpoint measurements and cannot capture the dynamic nature of adenylate metabolism in vivo. Emerging trends emphasize integrative, miniaturized, and minimally invasive strategies for near-real-time monitoring. In particular, in vivo solid-phase microextraction (SPME) has gained attention as a minimally invasive sampling technique capable of extracting labile metabolites from tissues under physiological conditions. By assessing advancements and challenges, this review highlights the evolution of adenylate determination and the need for tools compatible with dynamic, spatially resolved sampling. • ATP, ADP, AMP analysis provides insight into physiological and pathological states. • Chromatographic strategies for the analysis of phosphate-rich compounds. • Sample preparation approaches for labile ATP, ADP, and AMP under native conditions. • Methodological challenges for ATP, ADP, and AMP determination in biological samples. • In vivo SPME as a minimally invasive tool for spatiotemporal profiling of adenylates.