基于液质联用的复方龙胆酊中龙胆、草豆蔻及陈皮的定性定量研究

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

目的 建立基于液质联用的复方龙胆酊中龙胆、草豆蔻、陈皮的定性定量研究的方法。方法 采用Inertsil ODS-3色谱柱（4.6 mm×150 mm，5 μm），柱温40 ℃，流动相为乙腈-0.1%甲酸溶液；采用HPLC分离龙胆苦苷、山姜素、小豆蔻明和橙皮苷。分别采用电喷雾离子源正、负离子模式扫描，选择多反应离子监测方式（MRM）进行定性监测原料药的投料情况及有效成分的定量分析。结果 通过分析供试品与混合对照药材、对照品的一级质谱图的特征峰，判断该中成药是按处方工艺投料。方法学验证结果表明，龙胆苦苷、小豆蔻明、山姜素和橙皮苷依次分别在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）范围内线性关系良好；龙胆苦苷、山姜素、小豆蔻明和橙皮苷精密度依次分别为0.81%、0.48%、0.61%、1.55%，平均回收率依次分别为95.08%、93.28%、94.78%、95.04%。结论 该法快速简便、准确可靠、灵敏度高；适用于全面监测和控制复方龙胆酊中龙胆、草豆蔻、陈皮的投料情况及质量。

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）.