Abstract: A simple, rapid and sensitive method for the determination of alkaloids content in Phellodendron amurense R. was established. Alkaloids was ultrasonically extracted with 63% alcohol at 41℃ for 64 min, and determined by high performance liquid chromatography. The alkaloids were separated on a KYA HIQ sil C18 column 250 mm×4.6 mm, 5 μm, using acetonitrile-aqueous solution 3.4 g KH2PO4 and 1.7 g SDS in 1 000 mL H2O 1:1 as the mobile phase at flow rate 1.0 mL/min and detected at 345 nm. Contents of berberine, palmatine and jatrorrhizine in the root bark, stem bark and leaves of P. amurense R. were determined. The results showed that alkaloids contents in the root bark, stem bark and leaves were different. Contents of berberine and jatrorrhizine were higher in the root bark than in the stem bark, and they were the lowest in the leaves; while palmatine content was the highest in the stem bark, then in the root bark, and the lowest in the leaves too.