Simultaneous Analysis of Protoberberine and Indolequinoline Alkaloids in Zuojin Pill Extract by High-Performance Liquid Chromatography

As a new form of traditional Chinese medicine (TCM), Zuojin pill extract is made containing Coptidis rhizoma (the rhizome of Coptis chinesis Franch. [Ranunculaceae]) and Euodiae fructus (the unripe fruit of Euodia rutaecarpa [Juss.] Benth. [Rutaceae]) at the ratio of 6:1 (w/w) and has been most widely used in TCM to treat gastro-intestinal disorders. However, the quality control is insufficient. For establishing an analysis method for the effective quality control of Zuojin pill extract, the active components of three protoberberine alkaloids (coptisine, palmatine, berberine, components from C. rhizoma) and two indolequinoline alkaloids (evodiamine, rutaecarpine, components from E. fructus) in Zuojin pill extract were determined simultaneously by high-performance liquid chromatography. Chromatographic separations were performed on an Inertsil ODS-3 column (4.6 mm x 250 mm, 5 mu m). Elution was carried out at 25 degrees C under isocratic conditions by using 25 mmol L-1 KH2PO4-25 mmol L-1 SDS-acetonitrile (1:1:2, v/v/v, adjusted pH to 3.0 using phosphoric acid) as a mobile phase with a flow rate of 0.5 mL min(-1). Detection wavelength was set at 264 nm. Good linearity was obtained with correlation coefficient higher than 0.999 for all the analytes over the investigated concentration ranges. Three batches of Zuojin pill extract were successfully analyzed. The average percentages of coptisine, palmatine, berberine, evodiamine, and rutaecarpine in Zuojin pill extract were 5.35%, 3.92%, 16.64%, 0.03%, and 0.03%, respectively. Consequently, these protoberberine and indolequinoline alkaloids could be identified and determined easily by the established high-performance liquid chromatography (HPLC) method which can be used to evaluate and control the quality of Zuojin pill extract.