虽然远离完美,通过同时测定一组成分来评估给定草药(HM)的质量是可行的。然而,LC-MS/MS平台的定量结果的可信度存在几个技术壁垒,如化学降解,极性范围,浓度跨度,和身份错误识别。在这里,我们试图通过整合几种适合目的的技术来规避这些障碍,包括在线提取(OLE),串联反相LC-亲水相互作用液相色谱(RPLC-HILIC),定制的多反应监测(MRM),和相对反应与碰撞能量曲线(RRCEC)匹配。增强信心的肉馅盐(CSA)定量分析,一种著名的沙生植物和滋补草药,是作为概念验证进行的。部署OLE模块以禁止化学降解,特别是苯乙醇糖苷的E/Z-构型转化。由于连续通过RPLC和HILIC柱,无论极性如何,每种分析物都发生了令人满意的保留。次要组件的最佳参数,在丰富成分的劣质成分的同时,确保所有内容在线性范围内的位置。通过匹配保留时间来实现捕获信号的明确分配,离子跃迁,更重要的是,真实化合物和可疑峰之间的RRCEC。多种验证试验证明了新开发的方法是可靠的。特别是,尽管这些异构体显示出相同的保留时间和离子跃迁,但仍公开了甘露醇而不是半乳糖醇的分布。在Csa以及两个类似物种中确定了21种感兴趣的化合物的含量,数量模式不仅在不同物种之间而且在不同Csa样品之间都存在很大差异。一起,通过RRCEC匹配对OLE-RPLC-HILIC量身定制的MRM进行强化,可以完全满足HMs的置信度增强定量分析的需求。
Although far away from perfect, it is practical to assess the quality of a given herbal medicine (HM) through simultaneous determination of a panel of components. However, the confidences of the quantitative outcomes from LC-MS/MS platform risk several technical barriers, such as chemical degradation, polarity range, concentration span, and identity misrecognition. Herein, we made an attempt to circumvent these obstacles by integrating several fit-for-purpose techniques, including online extraction (OLE), serially coupled reversed phase LC-hydrophilic interaction liquid chromatography (RPLC-HILIC), tailored multiple reaction monitoring (MRM), and relative response vs. collision energy curve (RRCEC) matching. Confidence-enhanced quantitative analysis of Cistanche salsa (Csa), a well-known psammophytic species and tonic herbal medicine, was conducted as a proof-of-concept. OLE module was deployed to prohibit chemical degradation, in particular E/Z-configuration transformation for phenylethanoid glycosides. Satisfactory retention took place for each analyte regardless of polarity because of successive passing through RPLC and HILIC columns. Optimum parameters for the minor components, at the meanwhile of inferior ones for the abundant ingredients, ensured the locations of all contents in the linear ranges. The unequivocal assignment of the captured signals was achieved by matching retention times, ion transitions, and more importantly, RRCECs between authentic compounds and suspect peaks. Diverse validation assays demonstrated the newly developed method to be reliable. Particularly, the distribution of mannitol rather than galactitol was disclosed although these isomers showed identical retention time and ion transitions. The contents of 21 compounds-of-interest were definitively determined in Csa as well as two analogous species, and the quantitative patterns exerted great variations among not only different species but different Csa samples. Together, the fortification of OLE-RPLC-HILIC-tailored MRM with RRCEC matching could fully address the demands from confidence-enhanced quantitative analysis of HMs.