The equilibrium constants of five deoxyribonucleosides (dDyd, dUrd, dGuo, dThd, dAdo) were estimated by the plate theory and the moment method under isocratic conditions of the Reversed-Phase High Performance Liquid Chromatography (RP-HPLC). The mobile phase in this system was composed of water and organic modifiers(acetonitrile and methanol) The plate theory of linear adsorption isotherm was treated on the basis of continuous flow of eluent through the plates of the column. The moment method was utilized to find the equilibrium constant from the first absolute moment of experimental data. The equilibrium constants of five deoxyribonucleosides in the two methods were very close, and also the equilibrium constants calculated by capacity factor were similar to those by both the plate theory and the moment method. The equilibrium constant was expressed as a semi-log function of the quantity of organic modifier. Excellent agreements between the calculated elusion profile by the plate theory and the experimental data were observed.