High-performance liquid chromatographic techniques can be very effective for the resolution and isolation of nucleic acids. The characteristic ionic (phosphodiesters) and hydrophobic (nucleobases) properties of RNAs can be exploited for their separation. In this respect anion-exchange and reversed-phase chromatography have been successfully employed in the analysis and/or isolation of RNAs. In some cases, particularly tRNAs, chromatographic separations which rely upon multiple interactions between the solute and mobile and/or stationary phases have been highly effective. Mixed-mode chromatography involving simultaneous ionic and hydrophobic interactions, has been employed to resolve complex mixtures of tRNAs. Hydrophobic-interaction chromatography using gradients of decreasing salt concentration and weakly hydrophobic stationary phases has allowed the resolution of some tRNA mixtures as well as the analysis of modified materials.