Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China.
Anal Chim Acta. 2012 Feb 3;713:121-9. doi: 10.1016/j.aca.2011.10.062. Epub 2011 Nov 23.
A comprehensive two-dimensional (2D) separation is one that employs two separation dimensions (columns) and draws on all of the available resolving power from each of the dimensions of separate the components in a sample. In this study, a comprehensive 2D chromatography approach was developed for the separation and identification of membrane permeable compounds in a famous traditional Chinese medicine of Schisandra chinensis. The first dimensional column was the immobilized liposome chromatography (ILC) column, which mimics the biological membranes and can be used to study drug-membrane interactions in liquid chromatography. Using an automatic ten-port switching valve equipped with two sample loops, the section of the first-dimension was introduced in the second-dimension consist of a silica monolithic column. More than 40 components in Schisandra chinensis were resolved by using the developed separation system and among them 14 compounds were identified interacting with the ILC column based on their retention action, UV and mass data. With this comprehensive 2D-HPLC system, the three-dimensional chromatographic fingerprints of Schisandra chinensis were preliminarily established and processed by using principal component analysis and hierarchical clustering analysis. The obtained information can distinguish the unacceptable samples of the quality control. The result demonstrated that the 2D biochromatography system has been demonstrated to have more advantages of finding strong binding bioactive components, providing an enhanced peak capacity, good sensitivity and powerful resolution biological fingerprinting analysis of complex TCMs, which was a useful means to control the quality of and to clarify the membrane permeability of the compounds in Schisandra chinensis.
一种全面的二维(2D)分离是指采用两种分离维度(柱),并利用每个维度的所有可用分辨率来分离样品中的各个组分。在这项研究中,开发了一种全面的 2D 色谱方法,用于分离和鉴定五味子这一著名中药中的膜通透性化合物。第一维柱是固定化脂质体色谱(ILC)柱,它模拟生物膜,可用于在液相色谱中研究药物-膜相互作用。使用配备两个样品环的自动十通切换阀,将第一维的一部分引入由硅胶整体柱组成的第二维。使用开发的分离系统,可将五味子中的 40 多种成分分离,其中 14 种化合物根据保留行为、UV 和质谱数据被鉴定为与 ILC 柱相互作用。使用这种全面的 2D-HPLC 系统,初步建立了五味子的三维色谱指纹图谱,并通过主成分分析和层次聚类分析进行了处理。所得信息可区分质量控制的不合格样品。结果表明,2D 生物色谱系统在寻找强结合生物活性成分方面具有更多优势,提供了更高的峰容量、更好的灵敏度和强大的分辨率,用于分析复杂中药的生物指纹图谱,这是控制质量和阐明五味子中化合物的膜通透性的有用手段。