Key Laboratory of Ministry of Education for Traditional Chinese Medicine Resource and Compound Prescription, Hubei University of Chinese Medicine, Wuhan 430065, PR China; Key Laboratory of Particle & Radiation Imaging, Ministry of Education, Department of Engineering Physics, Tsinghua University, Beijing 100084, PR China.
China Pharmaceutical Preparation Section, Huazhong University of Science and Technology Union Jiangbei Hospital/Wuhan Caidian People's Hospital, Wuhan 430100, PR China.
J Chromatogr A. 2020 Dec 6;1633:461650. doi: 10.1016/j.chroma.2020.461650. Epub 2020 Oct 29.
For a successful high-speed countercurrent chromatography (HSCCC) separation of multiple components, the suitable solvent system selection is a critical operation. Despite the difference in separation mechanism between HSCCC and HPLC, K values of compounds in the solvent system and the retention factor of compounds in HPLC were connected with polarity, and the polarity mainly depended on the structure and chemical properties of compounds. On the basis of the concept of "like dissolves like", the average polarity of the solvent system is equal to the average polarity of multiple components at the "sweet point" log K = 0. The result of theoretical deduction showed a negative linear correlation between the polarity of each component and the logarithm of the gradient range of the organic phase. Therefore, the average polarity of unknown multiple components could be obtained through the polarity parameter model established in the HPLC analysis. The suitable solvent system composed of n-hexane-ethyl acetate-methanol-water (8:3:8:1, v/v) was selected through the average polarity of multi-components combining K values of the enriched samples. In the context of bioassay, an efficient HSCCC separation procedure was established, and two groups of analogue benzaldehyde derivatives (four main antioxidants, namely, isodihydroauroglaucin, isoaspergin, isotetrahydro-auroglaucin, and flavoglaucin; two minor antioxidants, namely, 6'-oxo-chaetopyranin and chaetopyranin) were obtained from Hypsizygus marmoreus. The predicted polarity values of multi-components were sufficient to meet the HSCCC experimental requirements. The HPLC analysis of reference compounds and multi-components showed a significant consistency with different chromatographic columns. Therefore, the polarity parameter model established in the HPLC analysis was a simple, rapid, and helpful tool for looking an appropriate solvent system, which was a forwarding step for the HSCCC separation of multiple components.
为了成功地进行高速逆流色谱（HSCCC）的多组分分离，合适的溶剂系统选择是一个关键操作。尽管 HSCCC 和 HPLC 的分离机制不同，但化合物在溶剂系统中的 K 值和化合物在 HPLC 中的保留因子与极性有关，而极性主要取决于化合物的结构和化学性质。基于“相似相溶”的概念，溶剂系统的平均极性等于“甜蜜点”（log K = 0）时多个组分的平均极性。理论推导的结果表明，每个组分的极性与有机相梯度范围的对数之间呈负线性相关。因此，通过在 HPLC 分析中建立的极性参数模型，可以获得未知多组分的平均极性。通过结合富集样品的 K 值，选择由正己烷-乙酸乙酯-甲醇-水（8:3:8:1，v/v）组成的平均极性合适的溶剂系统。在生物测定的背景下，建立了一种有效的 HSCCC 分离程序，并从杏鲍菇中获得了两组类似的苯甲醛衍生物（四种主要抗氧化剂，即异二氢金缕梅素、异麦角碱、异四氢金缕梅素和黄酮醇；两种次要抗氧化剂，即 6'-氧-壳聚糖素和壳聚糖素）。多组分的预测极性值足以满足 HSCCC 实验要求。参考化合物和多组分的 HPLC 分析显示出与不同色谱柱的显著一致性。因此，在 HPLC 分析中建立的极性参数模型是寻找合适溶剂系统的一种简单、快速和有用的工具，这是 HSCCC 多组分分离的一个前进步骤。
