Evaluation of Inter-Apparatus Separation Method Transferability in Countercurrent Chromatography and Centrifugal Partition Chromatography

Simon Roehrer,Mirjana Minceva

doi:10.3390/separations6030036

Simon Roehrer, Mirjana Minceva

Open Access

https://doi.org/10.3390/separations6030036

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Journal: Separations	Publication Date: Jul 24, 2019
Citations: 24	License type: CC BY 4.0

Affiliation: Technical University of Munich

Abstract

In the countercurrent chromatography and centrifugal partition chromatography, separation method transfer and scale-up is often described as an easy and straightforward procedure. Separation methods are usually developed on lab scale columns and subsequently transferred using linear scale-up factors to semi-preparative or preparative columns of the same column design. However, the separation methods described in the literature have been developed on various columns of different design and size. This is accompanied by differences in the separation behavior of the columns and therefore makes separation method transfer difficult. In the current study, the separation performances of different columns were evaluated and compared. Linear correlations of stationary phase retention and column efficiency as a function of flow rate were found to be applicable for the calculation of separation resolution in the typical operating range of each column. In this context, a two-point short-cut approach for a fast column characterization is recommended. This allows a quick prediction of the separation method transferability between columns, which saves experimental time and effort. In the current study, the transferability between five different columns from lab scale countercurrent chromatography (CCC) (18 mL) to semi-preparative centrifugal partition chromatography (CPCs) (250 mL) with different cell numbers and design is investigated.

Highlights

Solid support-free liquid-liquid chromatography (LLC) is a powerful technique with increasing popularity for the separation of active compounds from plant extracts and biotechnological products
Two fundamental column designs exist: hydrodynamic columns better known as countercurrent chromatography (CCC), and hydrostatic columns known as centrifugal partition chromatography (CPC)
The different column characteristics can be described by a different stationary phase retention, column efficiency and peak resolution

Summary

Introduction

Solid support-free liquid-liquid chromatography (LLC) is a powerful technique with increasing popularity for the separation of active compounds from plant extracts and biotechnological products. Two fundamental column designs exist: hydrodynamic columns better known as countercurrent chromatography (CCC), and hydrostatic columns known as centrifugal partition chromatography (CPC). CCC columns consist of a coil that is wound on a bobbin that rotates around its own axis and revolves around a second axis of a centrifuge [1,2]. CPC columns have only one axis of rotation with a constant centrifugal field. Conventional CPC columns consist of several identical annular disks that are connected in series, with polytetrafluoroethylene (PTFE) plates in between acting as seals. CCC columns, for example, differ in its rotary motion, ratios of the rotation radii (β-values), and different coil or tubing diameters and geometries, while CPC columns often have different cell and duct design as well as size.

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