The CE-Way of Thinking: "All Is Relative!".
Methods in molecular biology, January 2016
Philippe Schmitt-Kopplin, Agnes Fekete
Over the last two decades the development of capillary electrophoresis instruments lead to systems with programmable sampler, separation column, separation buffer, and detection devices comparable visually in many aspects to the setup of classical chromatography.Two processes make capillary electrophoresis essentially different from chromatography and are the basis of the CE-way of thinking, namely, the injection type and the liquid flow within the capillary. (1) When the injection is made hydrodynamically (such as in most of the found applications in the literature), the injected volumes are directly dependent on the type and size of the separation capillary. (2) The buffer velocity is not pressure driven as in liquid chromatography but electrokinetically governed by the quality of the capillary surface (separation buffer dependant surface charge) inducing an electroosmotic flow (EOF). The EOF undergoes small variations and is not necessarily identical from one separation or day to the other. The direct consequence is an apparent nonreproducible migration time of the analytes, even though the own velocity of the ions is the same.The effective mobility (field strength normalized velocity) of the ions is a possible parameterization from acquired timescale to effective mobility-scale electropherograms leading to a reproducible visualization and better quantification with a direct relation to structural characters of the analytes (i.e., charge and size-see chapter on semiempirical modelization).
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