Application of monolithic column in quantification of gliclazide in human plasma by liquid chromatography.

A simple, rapid and sensitive isocratic reversed phase HPLC method with UV detection using a monolithic column has been developed and validated for the determination of gliclazide in human plasma. The assay enables the measurement of gliclazide for therapeutic drug monitoring with a minimum quantification limit of 10ngml(-1). The method involves simple, one-step extraction procedure and analytical recovery was complete. The separation was carried out in reversed-phase conditions using a Chromolith Performance (RP-18e, 100mmx4.6mm) column with an isocratic mobile phase consisting of 0.01M disodium hydrogen phosphate buffer-acetonitrile (52:48, v/v) adjusted to pH 4.0. The wavelength was set at 230nm. The calibration curve was linear over the concentration range 10-5000ngml(-1). The coefficients of variation for inter-day and intra-day assay were found to be less than 6.0%.