Analysis of purine and pyrimidine bases,ribonucleosides, and ribonucleotides by high-pressure liquid chromatography

A high-pressure liquid chromatographic method has been developed for the separation and quantitation of purine and pyrimidine bases, ribonucleosides, and ribonucleotides. The procedure is carried out on a 1.8 × 700-mm column packed with Aminex-A-25 anion-exchange resin. The column is eluted with a linear gradient of ammonium chloride. The elution buffer contained borate also to complex the sugar phosphates and ethanol to improve the separation of bases and nucleosides. The analysis is completed in about 160 min. The potential application of this method for the quantitation of acid-soluble metabolites in fibroblasts is described.