Pharmacokinetic Study of Oxime Prodrug of Gliclazide by LC-MS/MS Method in Rabbit Plasma

Abstract

Oxime prodrug of gliclazide is a water soluble and biologically inactive derivative of gliclazide, a sulphonyl urea analogue used to treat type II diabetes mellitus. A rapid liquid chromatography tandem mass spectrometry LC-MS-MS method has been optimized for analysis of oxime prodrug of gliclazide in rabbit plasma using clopidogrel as internal standard. Following turboionspray ionization, the analytes were quantified on a triple– quadrupole mass spectrometer in multiple-reaction-monitoring (MRM) positive ion mode. Sample preparation involved a simple one-step protein precipitation with methanol, followed by centrifugation and evaporation of the organic solvent. The residue was redissolved in mobile phase and analyzed by LC–MS/MS. A Symmetry C18, 50x4.6, 5μ, a mobile phase composed of Acetonitrile: 25mM Potassium dihydrogen orthophosphate (pH6.5) (50:50 v/v), and a flow rate of 0.6 mL/min were employed, and the total run time was 3.0 min. The method was validated for accuracy, precision, linearity, selectivity, lower limit of quantification (LLOQ), recovery and matrix effect. The method was found to be linear in the range of 150 to 6000ng/mL. LLOQ was found to be 27ng/mL. All validation parameters met the acceptance criteria according to regulatory guidelines. This method was successfully applied to pharmacokinetic study of the prodrug in rabbit through oral administration.