Three monoterpenes were prepared,isolated and purified from Piper nigrum L..by high-speed counter-current chromatography(HSCCC).A two-phase solvent system composed of TBME-methanol-water(2∶1∶1,v/v/v) was used.The upper phase was stationary phase and lower phase was mobile phase.By one-step elution,under the conditions of a flow rate of 2.5 mL/min and 900 r/min,35 mg of 3,7-dimethyloct-2E-en-1,6,7-triol,55 mg of p-menthane-1,2,8-triol-2-O-β-D-glucopyranoside and 40 mg of 5-hydroxy bornyl-2-O-β-D-glucopyranoside was obtained from 2.76 mg of the butanol extract of Piper nigrum L.,the purities was over 95.0% detected by high performance liquid chromatography with evaporative light scattering detection.The obtained fractions were analyzed by high performance liquid chromatography(HPLC),and identified by mass spectrometry(MS),1H-nuclear magnetic resonance(NMR) and 13C-NMR.The results indicated that HSCCC is a powerful technique for the purification of monoterpenes from Piper nigrum L.with high productive capacity and high efficiency.