In this paper the separation of different mixtures of RNA derivatives by high-performance anion-exchange liquid chromatography is presented. By using this method different mixtures of nucleosides, 3#x2032;- and 5#x2032;- nucleotides and 2#x2032;:3#x2032;-cyclic nucleotides can be separated by isocratic elution at room temperature and in a short analysis time. The separation was optimized by properly adjusting the ionic strength and pH of the mobile phase. The pH is a critical factor in the order of elution and in the resolution capacity whereas the ionic strength affects primarily, the retention time. The separation method was applied to the analysis of RNAase A substrates and ligands.
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