高效液相色谱-二极管阵列检测/质谱法分析生脉饮煎剂中的人参皂甙类成分

Analysis of Ginsenosides in Sheng-Mai-Yin Decoction by High Performance Liquid Chromatography-Diode Array Detection-Electrospray Mass Spectrometry

采用高效液相色谱-二极管阵列检测/质谱(HPLC-DAD/M S)联用技术,以10mm o l/L醋酸铵和乙腈混合溶液梯度洗脱系统为流动相,应用C18色谱柱对生脉饮煎剂中人参皂甙类成分进行分离鉴定。分析结果表明:生脉饮煎剂中主要含有17个人参皂甙类成分,即20(R)-人参皂甙Rh1、Rh2、Rg3、Rg2,20(S)-人参皂甙Rh1、Rh2、Rg3、Rg2,人参皂甙R f、Rg6、Rg5、F4、Rk1、Rk3、Rh4,20(S)-和20(R)-原人参三醇。人参皂甙成分在煎煮过程中发生了很大变化,主要变成了一些中低极性产物,这是因为煎煮过程中发生了水解、差向异构、脱水等反应。该方法简便、精确、灵敏度高,可以用来分析生脉饮煎剂中人参皂甙的变化。

A method of high performance liquid chromatography coupled with diode array detection and electrospray ionization mass spectrometry （HPLC-DAD/MS） in negative ion mode was developed for the analysis of ginsenosides in Sheng-Mai-Yin decoction （Panax gingeng C. A. Mey, Ophiopogon japonicus （Thunb.） Ker-Gawl, Shisandra chinensis （Turcz.） Baill. ）. The analyses were preformed on a reversed-phase C18 column （4.6 mm i. d. × 150 mm, 5μm） using a binary eluent （10 mmol/L ammonium acetate （A） and acetonitrile （B）, 1 mL/min） under gradient conditions （60%A -40%A at 0 -30 min, 40%A -30%A at 30 -40 min）. Seventeen ginsenosides （20 （R）-Rh1, Rh2, Rg3, Rg2; 20（S）-Rh1, Rh2, Rg3, Rg2; Rf, Rg6, Rg5, F4, Rk1, Rk3, Rh4; 20 （S）- and 20 （R）-protopanaxatriol） were well separated and detected at 203 nm by a DAD detector. The effluent from the DAD detector was introduced into the electrospray ionization （ESI） source in a post-column splitting flow rate at 0.3 mL/min. In the mass spectrum two major ions [ M - H]^ - and [ M ＋ AcO ]^ - were observed for ginsenoside standards （20 （R）-Rh1, Rg3, Rh2; 20 （S）-Rh1, Rg3, Rh2; 20 （S）-and 20 （R）-protopanaxatriol） and ginsenosides in Sheng-Mai-Yin decoction. Some other ions [ M - Glc - H ]^ -, [ M - 2Glc - H ]^ -, [ M - Rha - H ]^ - and [ M - Rha - Glc - H ]^ - were also found in the mass spectrum of ginsenosides of Sheng-Mai-Yin decoction. In the decoction process ginsenosides changed into constituents of moderate and low polarity by hydrolysis, isomerization and dehydration at the site of C-20 and hydrolysis reaction also occurred at the site of C-3 or C-6. The work above presents a quick and accurate assay method which can could be used for the qualitative analysis of ginsenosides in Sheng-Mai-Yin decoction and the quality control of Sheng-Mai-Yin preparation.