Abstract

We develop a method on the basis of high-performance liquid phase chromatography–mass spectrometry (HPLC-MS) for determination of triterpene glycosides in ginseng extracts. In contrast to procedures based on HPLC with ultraviolet detection (HPLC-UV), commonly used for extract standardization, superior selectivity afforded by the developed method enables identification and quantitation of 23 major and minor ginsenosides. For this, in addition to using MS for highly selective detection of adduct ions composed of ginsenoside molecules and sodium and fragment sapogenin ions, we identify the conditions for chromatographic separation on a sorbent with grafted pentafluorophenyl groups. The effects that the temperature and mobile phase composition have on the selectivity of glycoside determination receive special attention here. For some pairs of compounds (F4/Rg6 and Rk3/Rh4), complete separation of chromatographic peaks is not achieved; nevertheless, even if present simultaneously, they can be determined owing to their different m/z ratios. The linearity ranges, equations for calibration curves, and analytical parameters (i.e., the limit of detection and reproducibility) are established for all analytes. The developed method is tested for standardization of reference extracts of Asian (P. ginseng) and American (P. quinquefolius) ginseng roots. For some ginsenosides, the content claimed by the manufacturer is at variance with the actual values, while for others the determined concentrations proved to be close to the claimed values. Additionally, we succeed in expanding the range of determinable ginsenosides, which is important for medical application of such extracts.

中文翻译：

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人参和西洋参根提取物的HPLC-MS标准化

摘要

我们开发了一种基于高效液相色谱-质谱（HPLC-MS）的方法，用于测定人参提取物中的三萜糖苷。与通常用于提取物标准化的基于HPLC的紫外检测程序（HPLC-UV）相比，该开发方法提供的优异选择性能够鉴定和定量23种主要和次要人参皂苷。为此，除了使用MS高度选择性地检测由人参皂甙分子，钠和皂苷元离子组成的加合物离子外，我们还确定了在具有接枝五氟苯基的吸附剂上进行色谱分离的条件。温度和流动相组成对糖苷测定选择性的影响在此受到特别关注。对于某些化合物对（F4 / Rg6和Rk3 / Rh4），无法完全分离色谱峰。但是，即使同时存在，也可以根据它们的不同来确定它们m / z比。建立所有分析物的线性范围，校正曲线方程式和分析参数（即检测限和重现性的极限）。测试了开发的方法，以对亚洲人参（P. quinquefolius）和美国人参（P. quinquefolius）人参根的参考提取物进行标准化。对于某些人参皂甙，制造商要求的含量与实际值存在差异，而对于另一些人参皂甙，确定的浓度被证明接近要求的值。此外，我们成功地扩大了可确定的人参皂苷的范围，这对于此类提取物的医学应用非常重要。