School of Pharmacy, Southwest Medical University, Luzhou, P. R. China.
J Sep Sci. 2021 Mar;44(5):973-980. doi: 10.1002/jssc.202001003. Epub 2021 Jan 15.
Preparative liquid chromatography has become an important purification method owing to its advantages of high separation efficiency, good reproducibility, and low solvent consumption. Because overloading in preparative liquid chromatography must be performed to increase the throughput in a cycle, nonlinear chromatographic behavior is observed. Therefore, it is crucial to carefully study nonlinear chromatography for the purification of a given product, which facilitates the efficient optimization of the purification parameters. In this work, a method for the development of a purification method using preparative liquid chromatography based on nonlinear chromatography is proposed. Hydroxytyrosol was selected as the subject for method demonstration. Using methanol and ethanol as organic modifiers, the optimum flow rate was determined on three commercial columns entitled C TDE, C ME, and C TDE, respectively. The curves were fitted with the van Deemter equation, with thorough analysis of the A, B, and C terms. Adsorption isotherms were subsequently studied to explore the distribution of solutes between the stationary and mobile phases at equilibrium. C TDE, 5 vol% ethanol-water, and 0.2 mL/min were selected as the optimal separation material, elution solvent, and flow rate, respectively. Purification of hydroxytyrosol was tentatively confirmed on a C TDE column with 1.6% sample loading, 90.98% recovery, and 98.01% purity.
制备液相色谱由于其具有分离效率高、重现性好、溶剂消耗低等优点，已成为一种重要的纯化方法。由于在制备液相色谱中必须进行过载以提高一个周期的通量，因此会出现非线性色谱行为。因此，仔细研究给定产物的非线性色谱对于纯化至关重要，这有助于高效优化纯化参数。在这项工作中，提出了一种基于非线性色谱的制备液相色谱纯化方法开发方法。以羟基酪醇为研究对象。分别以甲醇和乙醇为有机改性剂，在三种商用柱 C TDE、C ME 和 C TDE 上确定了最佳流速。用 van Deemter 方程对曲线进行拟合，对 A、B 和 C 项进行了详细分析。随后研究了吸附等温线，以探索在平衡时溶质在固定相和流动相之间的分布。选择 C TDE、5 vol%乙醇-水和 0.2 mL/min 作为最佳分离材料、洗脱溶剂和流速。在 C TDE 柱上进行羟基酪醇的初步纯化，样品加载量为 1.6%，回收率为 90.98%，纯度为 98.01%。
