Quantification of gliclazide by semi-micro high-performance liquid chromatography: Application to a bioequivalence study of two formulations in healthy subjects

Abstract

The objective of the present study was to evaluate the bioequivalence of two formulations of gliclazide in healthy human volunteers. Bioequivalence of the two formulations was determined in 20 healthy subjects with a single-dose, two-period, crossover study. A new high-performance liquid chromatographic method for the pharmacokinetic analysis of gliclazide was developed, using a semi-micro column to quantify gliclazide in plasma samples. Chromatographic separation was achieved with a semi-micro C18 column and 40 mM KH2PO4 (pH 4.6)-acetonitrile-isopropyl alcohol (5:4:1, v/v/v) as the mobile phase, and with UV detection at 229 nm. The method displayed good precision (coefficients of variation (CV<8.0%)), was fast (total analysis time 8 min), and required only a small amount of mobile phase (0.22 ml/min), with a reasonable limit of quantification (0.1 μg/ml). The calibration curve was linear in the concentration range 0.1-10 μg/ml. When the pharmacokinetic parameters of gliclazide in the two formulations were calculated and compared statistically using crossover analysis of variance, they were similar, with no statistically significant difference. Ninety percent confidence intervals for AUC0-last, AUC0-∞, and Cmax, used to evaluate bioequivalence, were in the stipulated range of 0.80-1.25. This result suggests that two formulations are bioequivalent when administered orally at a dose of 80 mg gliclazide. © 2004 Elsevier B.V. All rights reserved.