Abstract:This paper introduces the use of optimized solvent system with twice-ascending-development in twin trough chamber on TLC silica gel plate for the separation of protoberberine-type alkaliods eontained in Rhizomata of Coptis spp. Nine to eleven spotsincluding the main and the ‘minor‘ alkaloids in the samples can be observed on the chromatograms obtained under controlled conditions. The fluorescence and UV-absorption TLC scanning profile can serve as fingerprint for the analysis of commercial samples of Rhizomata of Coptis spp.