High resolution separation and quantitation of ribonucleotides using capillary electrophoresis

A method for the analysis of ribonucleotides using capillary electrophoresis has been developed. A cross-linked polyacrylamide coated column, Tris-HCl, and phosphate-mixed buffer were used, which produced reproducible separations (solute migration time average RSD% of 1.2) of 14 ribonucleotides within 50 min. Linear relationships between peak areas and sample concentrations, an average minimum detectable concentration of 5.4 microM, and an average minimum detectable quantity of 0.08 pmol were obtained. The described method allowed reproducible and reliable qualitative and quantitative analysis of intracellular ribonucleotide pools in HeLa cells.