High-performance frontal analysis for drug–protein binding study

Abstract

High-performance frontal analysis (HPFA) is a novel analytical method which enables simultaneous determination of total and unbound drug concentrations under drug–plasma protein binding condition. HPFA can be achieved using separation systems such as HPLC and CE. This paper deals with the principle and feature of HPFA method and its application to the stereoselective protein binding study. HPFA allows a simple analysis following direct sample injection, and does not suffer from undesirable drug adsorption on membrane nor leakage of bound drug through the membrane which are often encountered in conventional ultrafiltration and dialysis methods. HPFA can be easily incorporated into on-line HPLC system. By coupling HPFA with a chiral HPLC column, the unbound concentration of a racemic drug can be determined enantioselectively. The detection limit can be improved by coupling of HPFA with a preconcentration column. High-performance capillary electrophoresis/frontal analysis (HPCE/FA) enables to determine unbound concentrations enantioselectively with ultramicro injection volume, and is hence useful especially for the binding study of proteins which are scarce and difficult to obtain.