College of Pharmaceutical Science, Zhejiang University of Technology, Gongda Road 1, Huzhou 313000, China.
School of Forestry and Biotechnology, Zhejiang A and F University, Hangzhou 311300, China.
J Chromatogr A. 2022 Sep 13;1680:463422. doi: 10.1016/j.chroma.2022.463422. Epub 2022 Aug 14.
In this work, relationships between solvent strength of organic phase (ψ) for two biphasic solvent systems in high speed countercurrent chromatography, hexane-ethyl-acetate-methanol-water (HEMWat) and ethyl acetate-n-butanol-water (EBuWat), and partition coefficient (K) were investigated using four retention models, including Jandera's model (ABM), Neue-Kuss model (NK), linear-solvent-strength model (LSS) and quadratic-solvent-strength model (QSS). Experimental results showed that ABM model had the best fitting results for HEMWat system while NK model and QSS model had good fitting results in EBuWat system. Thus, a mathematical relationship between partition coefficient (K) and solvent strength of organic phase (ψ) could be obtained by measurement of partition coefficients of the target compounds with three different volume ratios of organic phase. At the same time, a functional map was proposed to construct to get a maneuverable region so that an optimal two-phase solvent system for separation of a target compound could be selected easily, which saved a lot of manpower for high speed countercurrent chromatographic separation. The application of this new method was declared by successful separation of two components, apigenin-6-C-β-D-xylopyranosyl-8-C-α-L-arabinopyranoside and vicenin-3, from dried leaves of Dendrobium officinale Kimura et Migo using high speed countercurrent chromatography.
在这项工作中，研究了两种双相溶剂体系高速逆流色谱中有机相溶剂强度（ψ）与分配系数（K）之间的关系，这两种双相溶剂体系为正己烷-乙酸乙酯-甲醇-水（HEMWat）和乙酸乙酯-正丁醇-水（EBuWat），使用了四种保留模型，包括 Jandera 模型（ABM）、Neue-Kuss 模型（NK）、线性溶剂强度模型（LSS）和二次溶剂强度模型（QSS）。实验结果表明，ABM 模型对 HEMWat 体系具有最佳的拟合效果，而 NK 模型和 QSS 模型对 EBuWat 体系具有良好的拟合效果。因此，可以通过测量目标化合物在三种不同有机相体积比下的分配系数，得到分配系数（K）和有机相溶剂强度（ψ）之间的数学关系。同时，提出了一个功能图来构建可操作区域，以便轻松选择用于分离目标化合物的最佳两相溶剂体系，这为高速逆流色谱分离节省了大量人力。通过使用高速逆流色谱成功分离铁皮石斛干叶中的两种成分，芹菜素-6-C-β-D-吡喃木糖基-8-C-α-L-阿拉伯吡喃糖苷和 vicenin-3，证明了该新方法的应用。
