Abstract: In this paper, a HPLC method was developed for the determination of alkaloids in Sophora flavescens Ait. Experimental evidences indicate that a system of a LiChrosorb--NH₂ column (4.0 mm×250 mm) as stationary phase and CH₃CN--H₃PO₄ (pH 2)--CH₃CH₂OH (80:8:10) as mobile phase can separate the five alkaloids, sophcarpine, matrine, sophoridine, oxysophcarpine and oxymatrine very well. This method is very easy and efficient and takes only 15 min for one run.