High Pressure Anion Exchange Chromatographic Measurement of Cyclic Adenosine 3′,5′-Monophosphate and Cyclic [14C]Adenosine Monophosphate Specific Activity in Myocardium Prelabeled with [14C]Adenosine

A 10-min high pressure pellicular anion exchange chromatographic method coupled to a new sensitive ultraviolet flow cell has been used to measure picomole amounts of cyclic adenosine 3',5'-monophosphate (cyclic AMP) in biological materials after previous purification of the samples. Tissue extracts in trichloroacetic acid are previously purified by anion exchange chromatography, zinc-barium precipitation, and thin layer chromatography. High specific activity cyclic [3H]AMP is used to determine purification recovery for each sample. The method has also been used to determine cyclic AMP and cyclic [14C]AMP specific activity in perfused frog ventricles prelabeled with [14C]adenosine. In addition, this method has been used to demonstrate that the positive inotropic effects of epinephrine and norepinephrine could not be dissociated from increased myocardial levels of cyclic AMP in the presence of the glycogenolytic blocking agent, N-isopropylmethoxamine.