Controlling of band width, resolution and sample loading by injection system in a simple preparative free-flow electrophoresis with gratis gravity.

J Chromatogr A

Laboratory of Analytical Biochemistry & Bio-separation, Key Laboratory of Microbiology of Educational Ministry, School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.

Published: April 2010

In this paper, the controllable band width, resolution and sample loading were investigated by the injection system of free-flow electrophoresis (FFE) with gratis gravity. Two general injection methods were described herein. The first method was the one in which sample injection fluxes were variable with constant background flux, while the second was the one in which the background fluxes were flexible with stable sample flux. With methyl green and crystal violet as two viewable model compounds, a series of experiments were performed, and the experimental results revealed that (1) the sample band width could be under desiring control through the regulation of ratios between sample and background fluxes, (2) the separative resolution could be also adjusted elaborately via the regulation of flux ratios during the separation of methyl green and crystal violet with only one charge disparity, and (3) the sample loading could be conveniently controlled via the flux ratios and an approximate maximum sample loading could be selected under the condition of just completed separation of two adjoining solutes. In addition, it was observed that the flux ratio had soft influence on the separative resolution of two solutes. These results were of significance to the designs on band width, resolution and sample loading in the newly developed FFE device with gratis gravity.

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http://dx.doi.org/10.1016/j.chroma.2010.01.085DOI Listing

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