An RP monolithic column coated with an amphoteric carboxybetaine type surfactant has been used with a combined triple eluent concentration, pH and flow gradient ion chromatography technique for the simultaneous separation of up to 18 nucleotides, nucleosides and nucleobases. The separation of up to eight precursors on a 1 cm long monolithic microcolumn using the combined gradient approach is also shown. The method was applied to the separation of the above nucleic acid precursors in perchloric acid extracts of yeastolates samples.