Sensitivity enhancement of capillary electrophoresis frontal analysis - based methods for characterization of drug - protein interactions by conductivity detection and preconcentration techniques
|Fakulta / Pracoviště MU
|Pharmacological research comprehends the study of drugs and plasma protein interaction. Plasma proteins do not act only as transport molecules but also as the reservoirs of drug molecules. Therefore, only the free fraction of the drug exhibits pharmacologic effect. The development of new methods to obtain as much information about binding as possible are in demand. Capillary electrophoresis (CE) technique is attractive for its speed of analyses, high efficiency, miniaturisation of analyses and possibility of automation. The capillary electrophoresis-frontal analysis (CE-FA) is established CE method for the affinity interaction studies. It allows the study in almost physiological conditions without labeling or immobilization of interaction partner. Despite of mentioned benefits there are certain limitation of the technique i.e. low concentration sensitivity associated with the mostly used UV-VIS detector. The aim of this study was to solve this problem by development new more sensitive methods based on the combination of CE-FA with conductivity detection or preconcentration techniques without an impact on the binding properties of the interaction. As the most abundant protein in the blood is serum albumin, this work was focused on its interaction with salicylic acid selected as model drug. Newly optimized methods have higher concentration sensitivity; estimated binding parameters are comparable with the data measured by conventional methods and with the data from the literature.