Rapid automated high-performance liquid chromatographic analysis of cyclic adenosine 3',5'-monophosphate. Synthesis...

A rapid and sensitive automated system for measuring cyclic adenosine 3',5'-monophosphate (cAMP) synthesized from either radiolabelled adenine or adenosine 5'-triphosphate (ATP) in intact and broken cell tissue preparations, respectively, is described. After incubation with radiolabelled precursor, tissue samples are deproteinized and then injected directly onto a reversed-phase high-performance liquid chromatographic column. The column effluent fraction which contains cAMP is collected into scintillation vials and assayed for tritium by liquid scintillation spectrometry. Since the high-performance liquid chromatographic and fraction collection procedures are automated, over fifty samples can be analyzed in duplicate in a single day. The utility of this assay is illustrated by investigations of the effects of beta-adrenergic receptor stimulation on cAMP synthesis in tissue slices prepared from rat cerebral cortex and dopamine on cAMP synthesis in striatal membrane preparations.