CAPILLARY ELECTROPHORESIS OF NUCLEOTIDES ON UCON-COATED FUSED-SILICA COLUMNS

Ucon-coated columns have been used to isolate nucleotides, such as rib onucleotides, deoxyribonucleotides, and pyridine nucleotides, by capil lary zone electrophoresis. This neutral, hydrophilic column coating (U con) significantly reduced macromolecule adsorption and electroosmotic flow, which provides the maximum resolution of the nucleotides separa ted under moderate buffer conditions (pH about 5-6), The relative stan dard deviations of the nucleotides were less than 1%. Low minimum dete ctable concentrations (about 2-8 mu M) and quantities (about 70-360 fm ol) of the ribonucleotides were obtained. A qualitative and quantitati ve analysis of the ribonucleotides in hybridoma cell extracts shows th e applicability of the developed methodology for the determination of intracellular nucleotide pools under the desirable buffer conditions. (C) 1994 Academic Press, Inc.