[Reevaluation of direct radioimmunoassay for plasma cAMP concentration]

In order to evaluate the validity of the direct radioimmunoassay of plasma cAMP level by using Yamasa cyclic AMP assay kit, we determined the cyclic AMP concentration of normal human plasma before and after purification through anion-exchange chromatography. We found that 1) the direct assay before purification gave almost the same values as the indirect assay after purification by anion-exchange chromatography, 2) hemolysis did not affect the results of the direct assay, 3) the plasma cAMP was stable for up to 12 weeks when 5 mM EDTA was added to the blood at the time of sampling and stored below -20 degrees C and 4) the cAMP concentration did not decrease by repeated freeze-thawing of the plasma samples for up to 10 times. These results indicate that the direct radioimmunoassay of plasma cAMP concentration in normal subjects can give the same values as those obtained after purification by anion-exchange chromatography. Although further investigation is needed to clarify whether these results can be equally applied to patient's plasma with various diseases, the direct assay offers a convenient and accurate method to determine plasma cAMP concentration.