Micellar liquid chromatography for the analysis of nucleosides and bases

A new high-performance liquid chromatographic method for the analysis of nucleosides and bases was developed in which a micellar mobile phase is used. Separation was achieved on a polyvinyl alcohol (PVA) column by isocratic elution with micellar sodium dodecyl sulfate (SDS) as the mobile phase. The retention behavior of the nucleosides and bases was significantly different from that obtained by reversed-phase chromatography. Effect of pH, temperature, and concentration of SDS and the counter ion (Na+) on retention behavior were investigated. With the PVA column, the best conditions for an isocratic separation were 0.01 M SDS (pH 3.4) and a flow-rate of 2 ml/min at ambient temperature. Mechanisms for the retention of the nucleosides and bases on the PVA column with a micellar mobile phase were proposed and an application of the separation was demonstrated by the analysis of human serum.