Reverse Phase High Performance Liquid Chromotography of the 2′- and 3′-Nucleotide Monophosphates

Abstract

A radial compression reverse phase column In conjunction with a high performance liquid chromotographic system has been developed for the separation of nucleotides, with special emphasis on the separation of the 2′- and 3′-nucleotide monophosphates which are produced by the base hydrolysis of RNA. The complete separation of the eight 2′- and 3′-isomers (as well as cyclic AMP) can be accomplished in twenty-five minutes using radial compression chemical bonded alkyl (C-18) reverse phase column by eluting with a reverse ionic strength gradient. Previous separation of these 2′- and 3′- isoraers required considerably longer time using the ion-exchange high performance liquid chro-matography methods.