An efficient method was developed for the preparative separation of palmatine and jatrorrhizine from Tinospora capillipes Gagnep.by pH-zone-refined counter-current chromatography(CCC). The two phase solvent system of chloroform-methanol-water(4∶3∶2) was selected,in which hydrochloric acid(20 mmol/L) was added into the upper aqueous phase as the stationary phase,and triethylamine(10 mmol/L) was added to the lower orgnic phase as the mobile phase.The apparatus was rotated at a speed of 850 r/min,while the mobile phase was pumped into the column at 2.0 mL/min.From 2 g of crude extract,512 mg palmatine and 421 mg jatrorrhizine with purities of 95.1% and 95.3% were obtained.The chemical identification of the isolated compounds was carried out by electrospray ionization-mass spectrometry(ESI-MS),1H NMR and 13C NMR.The introduced method is a rapid and efficient approach for the preparative separation of palmatine and jatrorrhizine in tinospora capillipes gagnep.