Pakistan Journal of Pharmaceutical Sciences Vol.15, No.2, July 2002, pp.51-56 METHOD DEVELOPMENT FOR ANALYSIS OF GLICLAZIDE IN HUMAN PLASMA BY USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY ROOHI OBAID, TASNEEM AHMED, OBAID ALI*, NOOR KAMIL* AND SYED WASEEMUDDIN AHMED** Research Institute of Pharmaceutical Sciences, Faculty of Pharmacy, University of Karachi *Department of Biochemistry, University of Karachi, **Department of Pharmacognosy, Faculty of Pharmacy, University of Karachi, Karachi-75270, Pakistan ABSTRACT Since 1970, dissolution requirements have been included in tablets and capsules monographs, in general, in response to concerns for bioavailability of equal significance is the recognition of the immense value of dissolution testing as a tool for quality control. Thus, equivalence in dissolution behaviour was sought in the light of both bioavailability and quality control considerations (Coppack et al., 1990). Nonetheless, dissolution profiles are often considered by the industry to ascertain the release rates of drug from tablet formulations as a quality assurance tool. However, in terms of sensitivity, precision and specificity, high-performance liquid chromatographic (HPLC) method may offer additional advantages (Charles & Ravenscroft, 1984 and Nawaz, 2001). INTRODUCTION Selective and sensitivie analytical method for quantitative determination of drugs and their metabolites are essential for successful evaluation of clinical pharmacology, pharmacokinetics (PK), bioavailability (BA) and bioequivalence (BE) studies (Raghow & Meyer, 1981). Gliclazide is known to possess low aqueous solubility (Nawaz, 2001). Large inter- and intra- individual responses following administration of sulphonylurea (glibenclamide etc.) reparations have also been reported (El-Sayed et al., 1989, Marchetti & Navalesi, 1989 and Charles & Ravenscroft 1984). Such variations are undesirable and may expose susceptible patients to the danger of hypoglycaemia or other associated hazards when a patient’s therapy is changed from one preparation to another. MATERIALS AND METHODS Potassium di hydrogen phosphate, guaranteed reagent tested according to International Standard Organization, Merck specification Acetonitrile were obtained from Merck and Reference Standard (RS) was obtained through the courtesy of Nabi-Qasim Pharmaceutical Industries. All stock solutions were prepared by dissolving RS in methanol followed by mobile phase/plasma. Chromatographic System: The chromatographic system used in this study consisted of a LC-5A, 6A system (Shimadzu- Japan) attached by UV. µ bonda pack C 18 300 mm x 3.9 mm internal diameter were used protected by Octa Decyl Silane guard column (pre column). A degassed and filtered mixture of Potassium Dihydrogen Phosphate (0.685 g in 1000 ml water) buffer pH 2.5 and Acetonitrile 30: 70 (v/v) was used as mobile phase. The flow rate was maintained at 1 ml/min. Detection was performed at 229 nm and separation was carried out at ambient temperature.