Chollet Sébastien, Marchal Luc, Renault Jean-Hugues, Legrand Jack, Foucault Alain
GEPEA, UMR CNRS 6144, Université de Nantes, 37 bd de l'Université 44602 Saint Nazaire Cedex, France; Kromaton, Rousselet Centrifugation SA, Annonay, France.
GEPEA, UMR CNRS 6144, Université de Nantes, 37 bd de l'Université 44602 Saint Nazaire Cedex, France.
J Chromatogr A. 2015 Apr 3;1388:174-83. doi: 10.1016/j.chroma.2015.02.043. Epub 2015 Feb 23.
Centrifugal Partition Chromatography (CPC) is a separation process based on the partitioning of solutes between two partially miscible liquid phases. There is no solid support for the stationary phase. The centrifugal acceleration is responsible for both stationary phase retention and mobile phase dispersion. CPC is thus a process based on liquid-liquid mass transfer. The separation efficiency is mainly influenced by the hydrodynamics of the phases in each cell of the column. Thanks to a visualization system, called "Visual CPC", it was observed that the mobile phase can flow through the stationary phase as a sheet, or a spray. Hydrodynamics, which directly governs the instrument efficiency, is directly affected during scale changes, and non-linear phenomena prevent the successful achievement of mastered geometrical scale changes. In this work, a methodology for CPC column sizing is proposed, based on the characterization of the efficiency of advanced cell shapes, taking into account the hydrodynamics. Knowledge about relationship between stationary phase volume, cell efficiency and separation resolution in CPC allowed calculating the optimum cell number for laboratory and industrial scale CPC application. The methodology is highlighted with results on five different geometries from 25 to 5000 mL, for two applications: the separation of alkylbenzene by partitioning with heptane/methanol/water biphasic system; and the separation of peptides by partitioning with n-butanol/acetic acid/water (4/1/5) biphasic system. With this approach, it is possible to predict the optimal CPC column length leading to highest productivity.
离心分配色谱法（CPC）是一种基于溶质在两个部分互溶的液相之间进行分配的分离过程。固定相没有固体载体。离心加速度负责固定相的保留和流动相的分散。因此，CPC是一种基于液-液传质的过程。分离效率主要受柱中每个单元内相的流体动力学影响。借助一种称为“可视化CPC”的可视化系统，观察到流动相可以以片状或喷雾状流过固定相。流体动力学直接决定仪器效率，在规模变化过程中会受到直接影响，并且非线性现象会阻碍成功实现掌握的几何规模变化。在这项工作中，基于考虑流体动力学的先进单元形状效率的表征，提出了一种CPC柱尺寸确定方法。关于CPC中固定相体积、单元效率和分离分辨率之间关系的知识，使得能够计算实验室和工业规模CPC应用的最佳单元数。该方法通过25至5000 mL的五种不同几何形状在两种应用中的结果得到突出展示：通过庚烷/甲醇/水双相体系分配分离烷基苯；以及通过正丁醇/乙酸/水（4/1/5）双相体系分配分离肽。采用这种方法，可以预测导致最高生产率的最佳CPC柱长度。
