Abstract
In this chapter, high-performance capillary electromigration (HPCE or CE) methods are presented as powerful tools for the determination of important physicochemical parameters of (bio)molecules. Using the different HPCE methods (zone electrophoresis in free solutions or in sieving/gel media, isotachophoresis, isoelectric focusing, affinity electrophoresis, and electrokinetic chromatography), the following physicochemical characteristics of a wide spectrum of compounds can be determined: effective, ionic and limiting mobilities, effective charges, isoelectric points, Stokes radii, relative molecular masses, partition constants, diffusion coefficients, and acidity (ionization) constants (pKa). In addition, the binding (stability, association, formation, or dissociation) constants of (bio)molecular complexes and rate constants of chemical and enzymatic reactions and noncovalent molecular interactions can be estimated. Advantages of HPCE methods for these physicochemical measurements include high separation efficiency, short analysis time, and especially the ultra-small mass (picogram range) and volume (nanoliter level) of injected solute samples and low consumption of chemicals and solvents. Moreover, the characterization of not quite pure solutes and solutes sparingly soluble in water is possible.
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