Simultaneous Determination of the Free and Total Carbamazepine Concentrations in Human Plasma by High-Performance Frontal Analysis Using an Internal-Surface Reversed-Phase Silica Support

Abstract

High-performance frontal analysis (HPFA) was applied to simultaneous determinations of the free and total carbamazepine (CBZ) concentrations in human plasma. When 1.5 ml of human plasma containing CBZ at a clinical therapeutic level (free fraction, about 30%) was directly injected into an internal-surface reversed-phase silica column, the CBZ was separated from the plasma bland and was eluted as a zonal peak with a plateau height corresponding to the free CBZ concentration in protein binding equilibrium. Slow and continuous introduction of the plasma sample and the use of titanium filters permitted us to inject the sample repeatedly while avoiding a rapid increase in column pressure. The free and total CBZ concentrations were determined simultaneously from the peak height at the plateau region and the area of the CBZ peak, respectively. The within-run and day-to-day reproducibilities were satisfactory (C.V.≤1.63%, n=5).