The static adsorption properties of alkaloids in nine kinds of macroporous resins were evaluated. The H103 macroporous resin which displayed the best adsorption capacity among the tested resins was selected out; the adsorption isotherm curve and adsorption kinetic behaviour of alkaloids with H103 macroporous resin were systematically studied, then the dynamic adsorption and desorption capability of alkaloids on H103 resin-packed column was investigated to establish optimal parameters for separation of matrine and oxymatrine. The different eluent was detected by thin layer chromatography (TLC), the sample of matrine or oxymatrine was detected by high performance liquid chromatography (HPLC). Results showed that the adsorption isotherm could be described by Freundlich isotherm equation; According to dynamics adsorption curve, the balance time was 100 minutes, Adsorption kinetic law could be expressed by Pseudo-second-order; the optimized dynamic process for gradient elution was 30% C2H5OH-25% NH4OH (115:1, v/v) and 80% C2H5OH, the oxymatrine was eluted firstly with recovery yield 85.3%, then matrine was eluted with recovery yield 90.1%, these two alkaloids could be separated effectively.