A new, simple method using ion-pair high-performance liquid chromatography was developed for simultaneous determination of berberine, palmatine and coptisine in Coptidis Rhizoma, Phellodendri Cortex and oriental pharmaceutical preparations containing them. A reversed-phase system of ODS-silica gel with a mixture of sodium lauryl sulfate, 0.1N tartaric acid, methanol and acetonitrile (0.5 : 49.5 : 10 : 40) as the mobile phase was used. The three components extracted with the mobile phase could be completely separated within 15 min. The detection limits for the three components were 15 ng at a signal-to-noise ratio of 5 : 1. The results obtained with an ultraviolet detector were in good accord with those obtained with a fluorescence detector.