Abstract: Tehandrine in three Chinese medicinal herb(Stephania terandra S.Moore,Stephania cepharantha Hayata,Menispermum dauricum DC)were firstly separated and determined by RP-HPLC and a chromatographic method for the separation and the determination of the tetrandrine was established. The chromatographic conditions are: ODS colunm, methanol-water-triethylamine(85:15: 0.25 V/v) mobile phase and UV detection at 283nm,This study offers a simple,convenient,sensitive and accurate method of separation and determination for expanding the exploition of tetrandrine medicament resources in Chinese medicinal herb and plants.