超临界流体色谱分离中药皂苷的色谱行为规律及特点研究

中药成分组成复杂，研究中面临一些分离、分析难题，如螺甾皂苷的C25-R/S异构体至今难以分离，中药高效液相指纹图谱存在一峰多个化合物等问题。超临界流体色谱（SFC)作为一种较新的分离分析手段，具有分离度高、选择性强、绿色环保等特点，近年在手性化合物分离等方面显示了优势，但其对中药成分的分离分析仅有零散报道，更没有对皂苷分离、分析的系统规律性研究和认识。本课题利用已经建立的皂苷化合物库（300余单体），观察皂苷分子中糖基、羟基、双键、酮基等的个数、连接位置对保留时间的影响，以及不同位置的立体异构体在SFC上的分离度；了解SFC分离皂苷的色谱行为规律及特点优势，以及与常规分离手段的互补性，形成SFC分离分析皂苷的一般流程及常规条件方法；从而指导SFC进行皂苷同分异构体的纯化制备及中药指纹图谱的分析表征，解决目前中药有效成分研究中所面临的分离分析难题，也为拓展SFC在中药研究中的应用奠定基础。

The composition of traditional Chinese medicine (TCM) is so complicated that there are some separation and analysis problems. For example, it is still difficult to separate some isomers such as C25-R/S spirostane saponins, and there are multiple compounds in one peak of some HPLC fingerprints of TCM. Supercritical fluid chromatography (SFC), a relative new separation and analysis technology, with the characteristics of high resolution, high selectivity, and friendly to environment, shows some advantages at the separation of chiral compounds. Till now, there are only a few reports on separation and analysis of the components of TCM by using SFC, and there is no systemic study and chromatographic behavior regularity of saponin separation by SFC. This proposal utilizes our established saponin library (over 300 compounds) to investigate effect of the numbers and position of glycosyl, hydroxyl, double bond and ketone group on retention time, and resolution of different saponin stereoisomers for SFC separation. In that case, we manage to understand the chromatographic behavior patterns and the characteristics of SFC separation of saponins, and the complementarity with conventional separation methods, and set up general work flow and routine conditions for separation and analysis of saponins on SFC. The results could guide the SFC usage on purification of saponin isomers and fingerprint analysis of TCM. SFC not only may solve the problem of separation and analysis of effective constituents, but also settle a foundation for expanding application of SFC in the research and development of TCM.