College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Jun 15;879(20):1773-80. doi: 10.1016/j.jchromb.2011.04.022. Epub 2011 Apr 22.
A novel method using column chromatography on oligo-β-cyclodextrin-Sepharose HP for the preparation of high purity daidzin from crude soybean samples was proposed in this work. The isoflavone of daidzin in sample A and B was purified under the optimum mobile phase composed of methanol/acetic acid/water=20.0/8.0/72.0 (v/v/v) at a flow-rate of 1.0 mL/min in one-step operation with a purity of 97.2% and 98.1%, a recovery of 95.3% and 96.3% respectively. The target products in isolated fraction were detected and characterized by HPLC analysis and ESI-MS spectrum. Preparative separation with sample-load of up to 2.42 mg/mL medium gave satisfactory results for daidzin with the purities over 97% and recoveries approximately 90%. Molecular docking simulations were utilized to help demonstrate the inclusion complexation between β-cyclodextrin and the isoflavones in samples through inclusion geometries and calculations of the binding energies. The prediction of the elution orders with AUTODOCK and SURFLEX-DOCK were validated by the chromatographic results.
本工作提出了一种使用寡-β-环糊精-琼脂糖 HP 柱层析从粗大豆样品中制备高纯度大豆苷元的新方法。在甲醇/乙酸/水=20.0/8.0/72.0（v/v/v）的最佳流动相下，在 1.0 mL/min 的流速下，一步法操作可将样品 A 和 B 中的大豆苷元异黄酮分别纯化至纯度为 97.2%和 98.1%，回收率分别为 95.3%和 96.3%。用 HPLC 分析和 ESI-MS 谱检测和表征分离馏分中的目标产物。在高达 2.42mg/mL 介质的样品负荷下进行制备性分离，可获得纯度超过 97%、回收率约为 90%的大豆苷元。分子对接模拟被用于帮助通过包合几何形状和结合能的计算来证明β-环糊精与样品中异黄酮之间的包合络合。AUTODOCK 和 SURFLEX-DOCK 的洗脱顺序预测通过色谱结果得到验证。
