Contactless conductivity detector as a tool for improving universality and sensitivity of capillary electrophoresis-frontal analysis: Proof of concept

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Authors

BRŽEZICKÁ Taťána MLČOCHOVÁ Hana GLATZ Zdeněk KOHÚTOVÁ Lenka

Year of publication 2024
Type Article in Periodical
Magazine / Source Journal of Separation Science
MU Faculty or unit

Faculty of Science

Citation
Web https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/jssc.202300667
Doi http://dx.doi.org/10.1002/jssc.202300667
Keywords binding constant; capillary electrophoresis-frontal analysis; contactless conductivity detection; human serum albumin; salicylic acid; C4D
Description Drug binding to plasma proteins influences processes such as liberation, adsorption, disposition, metabolism and elimination of drugs, which are thus one of the key steps of a new drug development. As a result, the characterization of drug protein interactions is an essential part of these time- and money-consuming processes. It is important to determine not only the binding strength and the stoichiometry of interaction, but also the binding site of a drug on a protein molecule, because two drugs with the same binding site can mutually affect free drug concentration. Capillary electrophoresis-frontal analysis with mobility shift affinity capillary electrophoresis is one of the most used affinity CE methods for the characterization of these interactions. In this study, a well-known sensitivity problem of most capillary electrophoresis-frontal analyses using UV detection is solved by its combination with contactless conductivity detection, which provided 6-fold lower limits of quantitation and detection. Binding parameters of the human serum albumin salicylic acid model affinity pair were evaluated by this newly developed approach and by the classical approach with UV detection primarily used for their mutual comparison. The results of both approaches agreed well, and are also in agreement with literature data obtained using different techniques.
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