Separation of major RNA-derived nucleotides by reversed-phase liquid chromatography

The effects of pH, ionic strength and amount of methanol in the eluent on the retention of 5'-, 3'- and 2'-ribonucleoside monophosphates on a reversed-phase high-performance liquid chromatographic system are described. The data were used to develop suitable separation protocols for synthetic nucleotide mixtures and applied to the separation of RNA nucleotides derived by alkaline hydrolysis.