Abstract
Micro high-performance frontal analysis (MHPFA) method has been developed for the simultaneous determination of unbound and total carbamazepine (CBZ) concentrations in protein binding equilibrium. When a 300-μl portion of 4-12μg/ml CBZ and 593μM human serum albumin (HSA) mixed solution was directly injected into a small bore ISRP column (10cm×2.0mm i.d.), CBZ gave a trapezoidal peak with a plateau separated from HSA. The CBZ concentration calculated from the plateau height agreed well with the unbound concentration determined by the conventional ultrafiltration-HPLC method, while the CBZ concentration calculated from the peak area coincided with the total (bound plus unbound) concentration. The within-run and day-to-day reproducibilities were satisfactory (relative standard deviation, <1.27%, n=5). Compared with the injection volume (1.4ml) required for the HPFA using a conventional size (15cm×4.6mm i.d.) ISRP column, MHPFA could reduce the sample size to about one-fifth (300μl).
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