Article | REF: P3365 V2

Capillary electrophoresis Principles

Author: Nathalie DELAUNAY

Publication date: December 10, 2019, Review date: September 2, 2020

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ABSTRACT

In capillary electrophoresis, the compounds are analyzed within a capillary with an internal diameter of a few tens of µm, filled with an electrolyte. The application of an electric field induces the movement of the compounds according to their electrophoretic migration and electroosmotic flow. This article presents the different parameters that influence these two phenomena, as well as the fundamental parameters (migration time, mobility, efficiency and resolution) that characterize the separation obtained. The different contributions to peak broadening are described. Finally, the different modes of separation are presented and illustrated by some applications.

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AUTHOR

  • Nathalie DELAUNAY: CNRS Researcher, Laboratory of Analytical Sciences, Bioanalytics and Miniaturization, UMR 8231 CBI, ESPCI Paris, France

 INTRODUCTION

Capillary electrophoresis is a technique for analyzing compounds in the liquid phase, be they small inorganic ions, small molecules such as amino acids, peptides or active pharmaceutical ingredients, or macromolecules such as proteins, polymers and DNA, or even nanoparticles or viruses. It is performed within a capillary of small internal diameter (a few tens of micrometers) filled with a separation electrolyte, and requires the application of an electric field. As with liquid chromatography, there are several separation modes, depending on the composition of the electrolyte and the possible presence of a stationary phase in the capillary, which enable compounds to be separated according to their charge/size ratio, hydrophobicity, chirality, size or isoelectric point, for example. This diversity of separation mechanisms, combined with very high efficiencies, rapid, automated, low-cost and miniaturized analyses, makes this technique indispensable in many applications. This article presents the different parameters influencing compound separation, defines the fundamental quantities and describes the various phenomena that can lead to a reduction in analysis performance. The principle of each separation mode in capillary electrophoresis is presented and illustrated by examples of various applications (inorganic ions, drugs, pollutants, proteins, DNA, etc.).

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KEYWORDS

separation   |   electrophoretic migration   |   electroosmotic migration


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Capillary electrophoresis