反相／亲水色谱法分析糖苷类化合物

Separation of Glucoside Compounds by Reversed-phase and Hydrophilic Interaction Liquid Chromatography

根据皂苷、黄酮苷等糖苷类化合物的结构特点,采用亲水色谱模式分析该类化合物,以弥补反相色谱模式分离结构类似糖苷类化合物选择性的不足。首先选择14个糖苷类化合物,比较了反相色谱柱(XAqua C18)和亲水色谱柱(Click XIon)的分离效果,评价了反相/亲水色谱的正交性,并构建了反相/亲水二维体系用于西洋参样品的分离。结果表明,糖苷类化合物在反相及亲水色谱柱上均有很好的保留,但两者具有不同的分离选择性,14种物质在反相和亲水柱的出峰顺序有较大差异,在反相色谱中不易分离的人参皂苷Rg1和Re在亲水色谱柱可获得很好的分离,反相/亲水模式分离糖苷化合物具有很好的正交性。以构建的RPLC/HILIC二维色谱体系分离西洋参样品,有效地提高了分离能力及峰容量,有利于后续更多极性及微量化合物的制备、结构表征与活性研究,且该方法操作简便、流动相兼容性好,可作为糖苷分离分析、制备的有效手段,也可以为其他中药复杂体系的分析提供参考。

Hydrophilic interaction liquid chromatography（HILIC） was selected to separate glucosides according to their structure characteristics. The insufficient of selectivity of reversed-phase chromatog- raphy（RPLC） would be made up. Firstly, the separation effects of 14 glucosides by both HILIC and RPLC columns were compared. The orthogonality was evaluated. Then the 2-D RPLC/HILIC system was used to separate American ginseng extract. The results showed that the glucosides were well re- served on both HILIC and RPLC columns. But the sequence of peaks was very different. Ginsen- oside Rgl and Re could not be separated with RPLC column, but they could be separated absolutely with HILIC column. The RPLC and HILIC modes had a high orthogonality. The peak capacity and separation ability were well increased for analyzing American ginseng extract by this 2-D RPLC/HILIC system. It was beneficial to the discovery of new compounds. This method was simple, and the mo- bile phase was compatible with both chromatographic modes. It could be used as the effective means for separation and preparation of glucosides. Also, it could provide a reference for the analysis of other complex traditional Chinese medicines.