Reversed-phase high-performance liquid chromatography (RP-HPLC) was used to determine the equilibrium isotherm of single and multi-components of dUrd(2'-deoxyuridine), dGuo(2'-deoxyguanosine), and dAdo(2'-deoxyadenosine) of 2'-deoxyribonucleosides by dynamic method. The composition of mobile phase was 90/10 vol.% (water/MeOH). With an increase in the injection volumes, the retention times were shorter and the peak shapes were triangleshaped, so Langmuir-type isotherm was assumed. The Langmuir adsorption parameters were estimated by PIM (pulsedinput method), and the competitive Langmuir adsorption isotherm was further utilized. For the sample of the dUrd and dGuo whose retention times were relatively short, the agreement of between the calculated value and experimental data was fairly good in both single and multi-components, but for the dAdo, the last eluting component, some deviations were caused by non-linear and non-ideal properties.