School of Pharmacy, East China University of Science and Technology, Shanghai, P R China.
J Sep Sci. 2012 Jul;35(14):1821-7. doi: 10.1002/jssc.201101103.
Two-dimensional reversed-phase liquid chromatography/hydrophilic interaction liquid chromatography (2D-RPLC/HILIC) system was successfully applied for comprehensive characterization of steviol glycosides from Stevia rebaudiana. The experiments were performed in offline mode using an XCharge C18 column in first dimension and an XAmide column in second dimension. In first dimension, preliminary separation of Stevia aqueous extract was accomplished and 30 fractions were collected. Then fractions 1-20 were selected for further purification and 13 compounds with high purity were obtained in second dimension. Comprehensive characterization of these compounds was completed by determination of their retention time, accurate molecular weight, diagnostic fragmentation ions, and nuclear magnetic resonance spectroscopy. As a result, all nine known steviol glycosides, as well as other four steviol glycosides were fully purified. The result demonstrated that this procedure is an effective approach for the preparative separation and comprehensive characterization of steviol glycosides in Stevia. This 2D-RPLC/HILIC method will be a promising tool for the purification of low-abundance compounds from natural products.
二维反相液相色谱/亲水相互作用液相色谱（2D-RPLC/HILIC）系统成功应用于甜菊叶中甜菊糖苷的综合特征分析。该实验采用离线模式，在第一维使用 XCharge C18 柱，在第二维使用 XAmide 柱。在第一维，初步分离甜菊叶的水提物，并收集 30 个馏分。然后选择馏分 1-20 进行进一步的纯化，在第二维得到 13 种高纯度的化合物。通过测定它们的保留时间、精确分子量、特征碎片离子和核磁共振波谱对这些化合物进行全面的特征分析。结果表明，所有 9 种已知的甜菊糖苷以及其他 4 种甜菊糖苷都得到了完全纯化。该方法为甜菊叶中甜菊糖苷的制备分离和综合特征分析提供了一种有效的方法。这种 2D-RPLC/HILIC 方法将成为从天然产物中纯化低丰度化合物的有前途的工具。
