Fast centrifugal partition chromatography was successfully applied in the separation of bioactive constituent niazirin directly from the chloroform extract of Moringa oleifera. The experiment was performed with a two-phase solvent system composed of ethyl acetate/BuOH/water (6:0.5:4v/v/v) in which the upper organic layer was used as stationary phase and lower aqueous phase was used as mobile phase. From 1g of crude extract, 70mg of niazirin was obtained in 94.8% purity as determined by HPLC. The total yield recovery was 94%. The isolated nitrile glycoside (niazirin) was characterized on the basis of its 1H, 13C-NMR and ESI-MS data.