Determination of the raw material feeding (Gentiana scabra, Katsumade Galangal Seed and dried tangerine peel) in compound gentian tincture by HPLC-MS/MS

Abstract

Objective To establish a HPLC-MS/MS method for comprehensive monitor and control of the raw material feeding (Gentiana scabra, Katsumade Galangal Seed and dried tangerine peel), and determination of Gentiopicroside, Alpinetin, Cardamonin and Hesperidin in Compound gentian tincture. Methods The separation was performed on an Inertsil ODS-3 (4.6 mm×150 mm, 5 μm) analytical column with the mobile phase consisting of acetonitrile-0.1% formic acid solution by gradient elution program, and the column temperature was 40 ℃. Active ingredients were separated by HPLC. The Electrospray Ionization Mass (ESI) source was applied and operated in the negative ion mode, and reactions ion monitoring mode (MRM) for quantitative analysis were selected. Results Through the analysis of the samples with mixed extract the same characteristic peak in MS was found to determine the proprietary Chinese medicine according to the prescription feeding process. The calibration curve of Gentiopicroside, Alpinetin,Cardamonin and Hesperidin were linear: 103.26-619.56 μg (r=0.999 0), 109.50-657.00 μg (r=0.999 5), 105.50-633.00 μg (r=0.996 9), 105.02-630.12 μg (r=0.999 5). The precision was Gentiopicroside 0.81%, Alpinetin 0.48%, Cardamonin 0.61% and Hesperidin 1.55% respectively. The average recovery rate were Gentiopicroside 95.08%, Alpinetin 93.28%, Cardamonin 94.78% and Hesperidin 95.04% respectively. Conclusions The method was proved to be simple, accurate, reliable, high sensitivity and can be used for determination and control of the raw material feeding (Gentiana scabra, Katsumade Galangal Seed and dried tangerine peel). Key words: Gentiopicroside; Alpinetin; Cardamonin; Hesperidin; Compound gentian tincture; HPLC-MS/MS