Int. J. Pharm. Sci. Rev. Res., 25(2), M ar – Apr 2014; Article No. 39, Pages: 207-210 ISSN 0976 – 044X International Journal of Pharmaceutical Sciences Review and Research Available online at www.globalresearchonline.net 207 Abeer Koul Ali 1 * , Ahmad Ali Hassan 2 1 *License in Pharmacy, Stage of preparing master degree in Pharmaceutical Chemistry and Quality control, Faculty of Pharmacy, Damascus University, Syria. 2 Professor at the Department of Pharmaceutical Chemistry and Quality control, Faculty of Pharmacy, Damascus University, Syria. * Corresponding author’s E-mail: abeerkoulali@yahoo.com Accepted on: 04-02-2014; Finalized on: 31-03-2014. ABSTRACT A simple, rapid, and precise reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed for the separation and simultaneous determination of Gliclazide, Glibenclamide, Glipizide and Glimepiride in bulk and pharmaceutical dosage forms. The elution was performed using a mobile phase mixture of methanol, acetonitrile and phosphate potassium buffer (pH= 3.5) in the ratio of 60:10:30 at a flow rate of 1.5 ml /min on a reverse phase C 18 column (150 × 4.6 mm, 5 μm) at 35 ̊ C. The drugs were monitored at a wavelength of 230 nm and were separated within 8 min. the method was validated as per ICH guidelines and the results were found to be within the acceptable range. Hence, the proposed method can be used for the routine quality control of the drugs. Keywords: Bulk and pharmaceutical dosage forms, RP-HPLC, Sulfonylureas, Validation. INTRODUCTION ulfonylureas were the first approved class of oral antidiabetics in the United States 1 , they were discovered accidentally in 1942. 2 They are classified as first-generation and second-generation agents. Second generation compounds, which include Gliclazide, glipizide, Glibenclamide and Glimepiride (Figure 1), are more potency and widely used in the treatment of type 2 diabetes. 3 Sulfonylureas stimulate insulin release from pancreatic β cells by binding to sulfonylurea receptors (SURs) on the β cell plasma membrane, which closes an adenosine triphosphate–dependent potassium ion (K + ) channels, leading to depolarization of the membrane and opening of Voltage-dependent calcium ion (Ca +2 ) channels, allowing influx of calcium ions and subsequent secretion of preformed insulin granules. 4-7 On the other hand, sulfonylureas act by reducing hepatic glucose production 5-7 and increasing peripheral insulin sensitivity 6,7 They are useful in the treatment of type 2 diabetes patient who can’t be managed by diet and increasing physical activity alone. 8, 9 Literature review assembled that a number of analytical methods have been reported for the determination of some antidiabetic sulfonylurea agents alone or with other antidiabetic drugs in human serum using HPLC-UV 10 or LC- MS 11 techniques, dosage forms using HPLC-UV 12,13 , or both of them using HPLC-UV. 14-16 One method has been developed for screening and determination of sulfonylurea agents in adulterated health food. 17 Although HPLC-UV was the most common technique used in these developed methods, HPLC methods using the most commonly available columns and inexpensive solvents were few. The aim of this study is to develop a rapid, sensitive, accurate and precise reverse phase HPLC method for the separation and simultaneous determination of Gliclazide (GLC), Glibenclamide (GLB), glipizide (GLP) and Glimepiride (GLM) in bulk and pharmaceutical dosage forms. The results of analysis were treated statically, as per International Conference on Harmonization (ICH) guidelines, for validation of analytical procedure. 18 The developed method would help in assay of drugs in a single run which reduces the time of analysis and doesn’t require separate method for each drug. In addition, all used solvents are inexpensive, so the developed method was economical and suitable for undeveloped areas. M ATERIALS AND M ETHODS Instrumentation The analysis of drug was carried out on a SHIM ADZU HPLC system equipped with a reverse phase C18 column (150x4.6mm, 5μm in particle size), a 20 μl injection loop and SPD-20A prominence UV/VIS detector, Degasser DGU-20A3. Reagents and solutions The reference standards of GLC, GLB, GLP and GLM were obtained from National Institute for the Control of Pharmaceutical and Biological Products in Syria HPLC grade water, methanol and acetonitrile were purchased from Merck Ltd., Mumbai (India). Potassium dihydrogen phosphate and ortho phosphoric acid were obtained from BDH Laboratory Supplies (Poole, UK). Chromatographic conditions Chromatographic separation of GLC, GLB, GLP and GLM was achieved on a reverse phase C18 column (150x4.6mm, 5μm in par cle size) at 35 ̊ C, in a mobile Development of a RP-HPLC M ethod for Simultaneous Determination of Some Antidiabetic Sulfonylurea Drugs in Bulk and Pharmaceutical Dosage Forms S Research Article