Application of simultaneous UV-radioactivity high-performance liquid chromatography to the study of intermediary metabolism. I. Purine nucleotides, nucleosides and bases.

Procedures are presented for the continuous-flow, simultaneous measurement of concentration and radioactivity of purine metabolites separated by high-performance liquid chromatography (HPLC). A microprocessor-controlled radioactivity flow detector connected in series to a UV-flow detector provides on-line quantitative monitoring of separated components in the post-column effluent stream. Through use of two HPLC separations--reversed-phase and anion-exchange--quantitation of all major purine nucleotides, nucleosides and bases is possible. The procedures provides a rapid, sensitive, and convenient means for the systematic study of purine metabolism.