Quantitative high-performance liquid chromatography of nucleosides and bases in human plasma

A reversed-phase high-performance liquid chromatographic (HPLC) method has been developed for the estimation of purines, pyrimidines and their congeners in biological fluids. An initial HPLC separation allowed the collection of a number of effluent fractions, each of which contained a single component of interest. The re-application of these fractions to a second HPLC separation permitted the resolution and quantification of nanogram amounts of these components. Isocratic elution with volatile buffers renders the samples amenable to automatic sampling procedures or lyophilisation. Data are presented on the application of the method to the analysis of nucleosides and bases in human plasma.