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2 Acta Chromatographica

2 2013

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Simultaneous analysis of protoberberine and indolequinoline alkaloids in Zuojin pill extract by high-performance liquid chromatography

Zhang H. M., Wang C. H., Wang Z. T.

Acta Chromatographica

2013

Vol. 25, no. 3

589--600

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 × 250 mm, 5 μm). Elution was carried out at 25°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.

Characterization and determination of trace alkaloids in seeds extracts from Peganum harmala linn. Using LC-ESI-MS and HPLC

Liu L., Zhao T., Cheng X. M., Wang C. H., Wang Z. T.

Acta Chromatographica

2013

Vol. 25, no. 2

221--240

Harmaline and harmine accounted for more than 70% in composition in extracts of P. harmala. More attention, however, should be paid to the other alkaloids which would be favorable or unfavorable to the efficacy and safety of the products. It was necessary to determine these trace alkaloids in the extracts; thereafter, most of them have been characterized. Diglycoside vasicine, vasicine, vasicinone, harmalol, harmol, tetrahydroharmine, 8-hydroxy-harmine, ruine, harmaline, and harmine were separated and identified with reference substances and characteristic MS spectra in extracts by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and high-performance liquid chromatography (HPLC). Three trace alkaloids, vasicine, harmalol, and harmol were determined using the developed chromatographic separation method subsequently. The average contents of vasicine, harmalol, and harmol in extracts of ten batches were 2.53 ± 0.73, 0.54 ± 0.19, and 0.077 ± 0.03%, respectively. The total content of the three alkaloids was 3.23 ± 0.90% (from 1.81 to 4.48%). For rough estimation of all the relative alkaloids except of harmaline and harmine, the average total areas of all peaks in extracts varied from 4.35 to 26.64% detected at 220, 254, 265, 280, and 380 nm, respectively. The results indicated that area normalization method was powerless for the quality evaluation for traditional herb medicine consisting of numerous compounds with highly differential features. It might be concluded that LC-MS or HPLC could be utilized as a qualitative and quantitative analytical method for quality control of the extracts from seeds of P. harmala L.