The Coptis Rhizome is known for containing a number of isoquinoline type alkaloids. Berberine, coptisine and palmatine are the major constituents of alkaloids. The alkaloids were isolated and determined by forming complex from Coptis japonica (Ranunculaceae). For the determination of these alkaloids, a new spectrophotometric method was developed with a simple and selective sample clean-up using thiocyanatocobaltate[II] complex ion. The absorbance of alkaloidal complex in l,2-dichloroethane solution was measured at 625 nm. A calibration curve for the alkaloids isolated from Coptis Rhizome was linear over the concentration range of 0.2-0.3 mg/mι. The method proved to be rapid, simple and reliable for the isolation and the determination of the alkaloids in Coptis Rhizome.