Behavior of Interacting Species in Capillary Electrophoresis Described by Mass Transfer Equation
journal contributionposted on 15.03.2006, 00:00 by Ning Fang, David D. Y. Chen
Affinity capillary electrophoresis (ACE) has been used to estimate thermodynamic constants of binding interactions with linear or nonlinear regression methods. The accuracy of this approach relies heavily on the binding interaction mechanism, which is controlled by both the nature of the interaction and the experimental conditions. The development of a highly efficient computer-simulated ACE system makes it possible to demonstrate the detailed behavior of any interacting species of a given interaction under any conditions. The order of the mobilities of the complex and the two binding species in their free forms is a key factor to determine what molecules in what locations of the column are involved in the interaction, and the peak shape resulting from such interactions, of a given ACE experiment. In this paper and the supporting materials, 18 scenarios in 6 different combinations of migration orders of the free analyte, free additive, and complex formed are studied by a computer simulation program based on the mass transfer equation. From the study of these situations, we conclude high additive concentration (ensuring high capacity factor) and low analyte concentration (ensuring fast fill-in of the free additive in the analyte plug) are crucial for obtaining accurate results when using the regression methods. On the other hand, the approach to estimate binding constants with computer simulation can be much more accurate as long as accurate and efficient simulation models can be developed, especially when the ratio of the additive and analyte concentrations is not large enough.
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simulation models18 scenariosCapillary Electrophoresis Describedanalyte plugMass Transfer Equation Affinity capillary electrophoresisACE experimentanalyte concentrationsbinding speciesmigration ordersnonlinear regression methodspeak shapecomputer simulation programadditive concentrationcapacity factorbinding interactionsanalyte concentrationregression methodsInteracting Speciesmass transfer equationbinding interaction mechanismcomputer simulationestimate binding constants