Open Access Research Article Analytical & Bioanalytical Techniques Ali et al., J Anal Bioanal Tech 2014, 5:5 http://dx.doi.org/10.4172/2155-9872.1000211 Volume 5 • Issue 5 • 1000211 J Anal Bioanal Tech ISSN: 2155-9872 JABT, an open access journal Keywords: Allopurinol; Benzbromarone; TLC-Densitometric; RP- HPLC Introduction 1,5-Dihydro-4H-Pyrazolo[3,4-d]pyrimidin-4-one (Figure 1) [1], known as Allopurinol (ALP), is an ofcial drug in British (BP) and United States (USP) Pharmacopoeias [1,2] which is used for treatment of gout and hyperuricaemia [3]. It is a xanthine oxidase inhibitor [4- 7], which prevents the oxidation of hypoxanthine to xanthine and xanthine to uric acid [8]. Tus results in the reduction of urate and uric acid concentrations in plasma and urine. 3,5-dibromo-4-hydroxyphenyl-2-ethyl-3- benzofuranyl methanone (Figure 2) [1], known as Benzbromarone (BENZ), is an ofcial drug in British (BP) Pharmacopoeia [1] which is used as a hypouricaemic drug. It increases the excretion of uric acid by blocking renal tubular reabsorption and thus reduces plasma concentrations and increases the excretion of uric acid [9,10]. Combination of ALP and BENZ has the advantages of greater therapeutic efect than with either drug alone [11]. Tis combination causes manifold reduction in uric acid concentrations in plasma and urine as compared to double dose of the individual drug when used alone [12]. Also, this combination helps to decrease the dose of each active ingredient, and as a result, decreases side efects of each component if given separately in high doses [13]. Reviewing the literature in hand, only one report has been published for determination of the studied mixture which depended on measuring BENZ using zero order spectra at its λ max =356 while ALP was determined by using ( 2 D) amplitudes at 281.4 nm or by measuring the amplitudes of the second derivative of the ratio spectra curves (2DD) at 282.4 nm afer using a standard spectrum of 8 μgmL -1 *Corresponding author: Weam Mahmoud Osman, Analytical Chemistry Department, Faculty of pharmacy, Misr University for Science and Technology (MUST), Egypt, Tel: 01004630203- 01117236884; E-mail: assem.wm@gmail.com Received September 15, 2014; Accepted October 16 2014; Published October 20, 2014 Citation: Ali NW, Abdelwahab NS, Fatatry HMEL, Osman WM (2014) Development and Validation of Different Chromatographic Methods for Determination of Two Hypouricemic Drugs in Their Combined Dosage Form. J Anal Bioanal Tech 5: 211 doi:10.4172/2155-9872.1000211 Copyright: © 2014 Ali NW, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Two sensitive, selective and precise chromatographic methods have been developed, optimized and validated for Allopurinol (ALP) and Benzbromarone (BENZ) determination in their pure form, laboratory prepared mixtures and pharmaceutical dosage form. The frst developed method was based on thin layer chromatographic (TLC) combined with densitometric determination of the separated spots. The separation was achieved using silica gel 60F 254 TLC plates. The mobile phase used was chloroform: methanol (9.2:0.8, v/v) and UV detection at 240 nm. Good correlations were obtained between the integrated peak area of the studied analytes and their corresponding concentrations in different ranges. The second developed method was based on the high-performance liquid chromatography with ultraviolet detection, by which the proposed components were separated using Zorbax C 18 column using a mobile phase consisting of sodium acetate buffer (pH=4.5, adjusted with acetic acid): acetonitrile : triethylamine (50:50:0.5, by volume) maintaining the mobile phase fow rate at 1 mLmin -1 with UV detection at 260 nm. Different parameters affecting the suggested methods were optimized for maximum separation of the cited components. System suitability parameters of the two developed methods were also tested. Validation of the methods has been carried out according to USP requirements and ICH guidelines, accuracy, precision and repeatability were found to be within the acceptable limits. The results obtained by TLC-Densitometric and RP-HPLC methods were statistically compared with those obtained by the reported RP-HPLC method and no signifcant difference was found regarding both accuracy and precision. Development and Validation of Different Chromatographic Methods for Determination of Two Hypouricemic Drugs in Their Combined Dosage Form Nouruddin W Ali 1 , Nada S Abdelwahab 1 , Hamed M EL Fatatry 2 and Weam M Osman 2 * 1 Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Bani-sueif University, Egypt 2 Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Misr University for Science and Technology (MUST) 6th October, Egypt (1H-pyrazolo [3,4-d]pyrimidin-4-ol)[1] Figure 1: Allopurinol (ALP) structure. (3,5-dibromo-4-hydroxyphenyl)-(2-ethyl-3- benzofuranyl) methanone Figure 2: Benzbromarone (BENZ) structure.