Wang Zhanliang, Wang Hong, Chen Shizhong
Research Laboratory of Traditional Chinese Medicine, School of Pharmaceutical Sciences, Peking University, Beijing 100083, China.
Se Pu. 2006 Jul;24(4):325-30.
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. x 150 mm, 5 microm) 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.
建立了一种高效液相色谱-二极管阵列检测-电喷雾电离质谱联用（HPLC-DAD/MS）负离子模式分析生脉饮（人参、麦冬、五味子）中人参皂苷的方法。分析在反相C18柱（内径4.6 mm×150 mm，5μm）上进行，采用二元流动相（10 mmol/L醋酸铵（A）和乙腈（B），1 mL/min），梯度洗脱（0～30 min，60%A～40%A；30～40 min，40%A～30%A）。17种人参皂苷（20（R）-Rh1、Rh2、Rg3、Rg2；20（S）-Rh1、Rh2、Rg3、Rg2；Rf、Rg6、Rg5、F4、Rk1、Rk3、Rh4；20（S）-和20（R）-原人参三醇）得到良好分离，DAD检测器在203 nm处检测。DAD检测器流出物以0.3 mL/min的柱后分流流速引入电喷雾电离（ESI）源。在质谱中，人参皂苷标准品（20（R）-Rh1、Rg3、Rh2；20（S）-Rh1、Rg3、Rh2；20（S）-和20（R）-原人参三醇）以及生脉饮中的人参皂苷观察到两个主要离子[M-H]-和[M+AcO]-。生脉饮人参皂苷质谱中还发现了一些其他离子[M-Glc-H]-、[M-2Glc-H]-、[M-Rha-H]-和[M-Rha-Glc-H]-。在煎煮过程中，人参皂苷通过C-20位的水解、异构化和脱水转变为中低极性成分，C-3或C-6位也发生水解反应。上述工作提供了一种快速准确的测定方法，可用于生脉饮中人参皂苷的定性分析和生脉饮制剂的质量控制。
