Determination of saikosaponin derivatives in Radix bupleuri and in pharmaceuticals of the Chinese multiherb remedy Xiaochaihu-tang using liquid chromatographic tandem mass spectrometry
Determination of saikosaponin derivatives in Radix bupleuri and in pharmaceuticals of the Chinese multiherb remedy Xiaochaihu-tang using liquid chromatographic tandem mass spectrometry
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DOI：
10.1021/ac0499423
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发表时间：
2004-07-15
影响因子：
7.4
通讯作者：
Nan, FJ
中科院分区：
文献类型：
--
作者：
Bao, YW;Li, CA;Nan, FJ
柴胡皂苷是从中草药柴胡中提取的具有生物活性的齐墩果烷型皂苷。建立了一种基于LC-MS/MS的方法，用于在一次色谱运行中表征和定量15种柴胡皂苷衍生物（柴胡皂苷a、柴胡皂苷B（1）、柴胡皂苷g、柴胡皂苷元A、柴胡皂苷元H、柴胡皂苷元c、柴胡皂苷元h、柴胡皂苷元i、原柴胡皂苷元C-2、原柴胡皂苷元B-2、柴胡皂苷元C、柴胡皂苷元B、柴胡皂苷元d、柴胡皂苷元B（2）和柴胡皂苷元D）。电喷雾和大气压化学电离技术在正离子和负离子模式下进行电离过程的优化。采用负离子ESI产生前体去质子化分子，以获得分析物的最佳电离灵敏度。此外，为每种分析物选择最丰富的碎片离子，以获得最佳CID灵敏度。由于一些皂苷衍生物是异构体，因此通过色谱和质谱实现了整个分析物的完全分离。此外，通过使用组合化学方法，成功地发现了用于测定分析物的最佳内标。在1.65或4.98 - 1200 ng/mL范围内观察到良好的线性关系。标称低、中和高浓度下的日内准确度和精密度分别在0.8 - 11.8%和80 - 116%之间变化，而日间含量测定的准确度和精密度分别在1.1 - 15.5%和86 - 119%之间变化。供试化合物的定量下限类似于16.5至49.4 pg柱上。新方法提供了更高的灵敏度和更大的特异性比以前报道的LC方法。验证后，这些化学品存在于各种粗柴胡根和不同品牌的中国多草药治疗小柴胡汤（或Shosaiko-to）提取物颗粒的测定方法的适用性已被证明。该技术的灵敏度和特异性将为准确定量生物基质中柴胡皂苷衍生物的方法奠定基础。
Saikosaponins are bioactive oleanane saponins derived from the Chinese medicinal herb Radix bupleuri ("chaihu" in Chinese). An LC-MS/MS-based method has been developed for characterization and quantification of 15 saikosaponin derivatives (saikosaponin a, saikosaponin b(1), saikosaponin g, saikogenin A, saikogenin H, saikosaponin c, saikosaponin h, saikosaponin i, prosaikogenin C-2, prosaikogenin B-2, saikogenin C, saikogenin B, saikosaponin d, saikosaponin b(2), and saikogenin D) in one chromatographic run. Optimization of the ionization process was performed with electrospray and atmospheric pressure chemical ionization techniques in both positive and negative ion modes. Negative ion ESI was adopted for generation of the precursor deprotonated molecules to achieve the best ionization sensitivity for the analytes. In addition, the most abundant fragment ion was chosen for each analyte to give the best CID sensitivity. Because some of the saponin derivatives are isomeric, complete resolution for the whole analytes was achieved both chromatographically and mass spectroscopically. Furthermore, optimal internal standard was successfully discovered for determination of the analytes by making use of a combinatorial chemistry approach. Good linearity over the range similar to1.65 or 4.98 to 1200 ng/mL for the analytes was observed. The intraday accuracy and precision at nominal low, intermediate, and high concentration varied between 0.8 and 11.8% and between 80 and 116%, respectively, whereas those for interday assay were between 1.1 and 15.5% and between 86 and 119%, respectively. The lower limits of quantitation for the test compounds were similar to16.5 to 49.4 pg on-column. The new method offered higher sensitivity and greater specificity than previously reported LC methods. After the validation, the applicability of the method for determination of these chemicals present in a variety of crude chaihu roots and in different brands of the Chinese multiherb remedy Xiaochaihu-tang (or Shosaiko-to) extract granules has been demonstrated. The sensitivity and specificity of the technique will be the basis of a method for the accurate quantification of the saikosaponin derivatives in biomatrixes.