OBJECTIVE:To establish a method for the simultaneous content determination of berberine hydrochloride, phellodendrine hydrochloride and magnoflorine in Phellodendron amurense decoction piece, and to campare the contents of the 3 ingredients in different grades of P. amurense decoction piece. METHODS: HPLC was performed on the column of Phenomenex Luna C18 with mobile phase of acetonitrile-0.05 mol/L KH2PO4 (gradient elution) at a flow rate of 1 ml/min, the detection wavelength was 280 nm, the column temperature was 30 ℃, and injection volume was 5 μl. RESULTS:The linear ranges were 0.387 0-7.740 μg for berberine hydrochloride (r=0.999 9),0.044 4-0.888 0 μg for phellodendrine hydrochloride (r=0.999 8) and 0.048 0-0.960 0 μg for magnoflorine(r=0.999 9); RSDs of precision, stability and reproducibility tests were lower than 3%, recoveries were 95.61%-103.22% (RSD=2.80%,n=6), 96.18%-102.80%(RSD=1.84%,n=6) and 97.93%-102.78% (RSD=1.84%,n=6). CONCLUSIONS:The method is simple and accurate, and can be used for the contents determination of berberine hydrochloride,phellodendrine hydrochloride and magnoflonine in P. amurense. The contents of berbenine hydrochloride and phellodendrine hydrochloride in the first-grade decation piece are higher than those in the second-grade decoction piece,and the content of magnoflorine in both decoction pieces shows no discernible differences.