Rapid separation of DNA constituents, bases, nucleosides and nucleotides, under the same chromatographic conditions using high-performance liquid chromatography with a reversed-phase column.

Four components of three sets of DNA constituents, bases, deoxyribonucleosides and deoxyribonucleoside 5'-monophosphate, were sufficiently resolved under one set of chromatographic conditions using high-performance liquid chromatography with a reversed-phase column (Zorbax ODS) and the solvent 0.4 M NH4H2PO4 (pH 3.5). The effect of pH and salt concentration in the solvent on the retention of these compounds in the column was thoroughly investigated. Proportionality of peak height to the content, and reproducibility and recovery of the four bases were satisfactory under appropriate conditions and as little as 1 microgram of DNA could be analysed for its base composition by this method.