Mixed mode chromatography via column switching for the simultaneous HPLC analysis of lonic and non-lonic nucleic acid constituents

Summary A method is presented for the separation of purine and pyrimidine nucleotides, nucleosides and bases in one analysis. This method uses both the ion-exchange and the reversed-phase modes for the analysis of these compounds. The modes are coupled through automatic switching valves. The use of these valves permits the isolation of either column during the analysis. The nucleotides are separated on an anion-exchange column and the nucleosides and bases on a C-18 column. Total analysis time is approximately one hour, and a very small volume of a sample can be analyzed. The technique is flexible and mobile phase conditions for one column can be altered without affecting the resolution on the other column. The method is applicable to biological matrices.