An assay of deoxyadenosine and adenosine in human plasma by HPLC

Abstract

We have developed a method for the simultaneous measurement of deoxyadenosine and adenosine in human plasma. A barium hydroxidezinc sulfate extraction procedure circumvented acid precipitation and preserved the deoxyadenosine. Deoxyadenosine and adenosine were separated using reverse-phase high-pressure liquid chromatography and their identities were confirmed by the enzymatic peak shift technique. A radioactive internal standard was used to measure extraction efficiency and to prove completeness of enzyme treatment for the peak shift technique. This assay is sensitive to 100 n of either nucleoside in plasma and can be completed within 1 hr. This technique can be used to quantitate plasma deoxyadenosine and adenosine in patients with congenital adenosine deaminase deficiency or in patients being treated with ADA inhibitors.