Determination of ATP, ADP, and AMP levels by reversed-phase high-performance liquid chromatography in cultured cells

Abstract

Cytoplasmic and mitochondrial Ca signals couple cellular ATP production to activity-related energy demand. In order to accurately determine the bioenergetic effect of Ca signals, cellular energy charge, i.e., the compound ratio of the phosphorylated adenine nucleotides AMP, ADP, and ATP, should be estimated. Reversed-phase high-performance liquid chromatography (RP-HPLC) allows the rapid separation and quantitation of these molecules. Here we describe a protocol applied in our laboratories to quantify ATP, ADP, and AMP nucleotides in cellular extracts.