Research Article A VALIDATED RP-HPLC METHOD FOR THE DETERMINATION OF SPARFLOXACIN IN BULK AND PHARMACEUTICAL DOSAGE FORM P. RAVISANKAR 1* , CH. DEVADASU 1 , G. DEVALA RAO 2 , M. NAGESWARA RAO 3 1 Department of Pharmaceutical Analysis and Quality Assurance, Vignan Pharmacy College, Vadlamudi, Guntur, A.P., India., 2 Department of Pharmaceutical Analysis, KVSR Siddhartha College of Pharmaceutical Sciences, Vijayawada, A.P., India., 3 Department of freshman engineering, Prasad V. Potluri Siddhartha Institute of Technology, kanuru, Vijayawada, A.P., India. Email : banuman35@gmail.com Received: 1 December 2013, Revised and Accepted: 26 December 2013 ABSTRACT Objective: To develop a simple, accurate, precise and selective isocratic reversed phase-high performance liquid chromatography (RP-HPLC) method was developed for the quantitative estimation of Sparfloxacin in pharmaceutical formulations. Experimental: RP-HPLC method was developed by using Welchrom C18 Column with 250 mm x 4.6 mm i.d and 5µm particle size, SHIMADZU LC- 20AT Prominence liquid chromatograph. The mobile phase used is phosphate buffer (pH-3.0): acetonitrile (70:30% v/v) with a flow rate of 1mL/min. The eluent was monitored by UV detection at 291nm using SHIMADZU SPD-20A prominence UV-Vis detector. Results: The retention time of Sparfloxacin found to be 5.499 min. The method retains linearity in the range of 2-10µg/mL with correlation coefficient 0.9999. The mean recovery of sparfloxacin was 99.702% to 100.456%. The LOD and LOQ of the drug were found to be 0.186 µg/mL and 0.558 µg/mL respectively. Conclusion: The proposed method can be successfully used for routine determination of sparfloxacin in pharmaceutical formulations. Keywords: Sparfloxacin, Isocratic RP-HPLC, Correlation coefficient , Method Validation. INTRODUCTION: The new fluoroquinolone Sparfloxacin (SPF) is (5-Amino-1- cyclopropyl-7-(cis-3,5-dimethyl-1-piperazinyl)-6,8-difluoro-1,4 dihydro-4-oxo-3-quinolinecarboxylic acid[1] (Figure 1), is a broad spectrum fluorinated a quinolone antibiotic used in the treatment of bacterial infections and commonly prescribed for infective opthalmitis and sinusitis, acute exacerbation of chronic bronchitis, community-acquired pneumonia, eye infections, urinary tract infection . SPF is a new difluorinated quinolone with similar activity for gram-negative and gram-positive bacteria and a spectrum of activity that embraces anaerobes, Chlamydiatrachomatis, Mycoplasma and mycobacteria [2]. The quinolones and SPF compounds are bactericidal in nature. The molecular target of quinolones is considered to be DNA gyrase, since quinolones inhibit gyrase activities and gyrases isolated from quinolone-resistant strains are resistant to quinolones [3-4]. Escherichia coli gyrase consists of subunits A and B which are the products of the gyrA and gyrB genes, respectively [5-7] since the unexpected finding by Shen and Pernet that [3H] norfloxacin binds to DNA but not to purified gyrase [8] it has been proposed that SPF exerts its antibacterial activity by inhibiting DNA gyrase which is a bacterial topoisomerase. DNA gyrase [9] is an essential enzyme which controls DNA topology and assists in DNA replication, repair, deactivation and transcription. N COOH O NH 2 F F HN H 3 C CH 3 Figure 1: Structure of SPF Literature survey revealed that very few methods have been reported for the analysis of SPF which includes luminescence spectroscopy, Reverse Phase High Pressure liquid Chromatography [10], RP-HPLC with fluorescence detection [11-18] and few UV spectrophotometric methods [19]. However most of the available RP- HPLC methods have limitations such as long run times, low sensitivity, uneconomical and have poor symmetry. Keeping in view of these we have decided to develop a simple, accurate, precise and reliable RP-HPLC method for the estimation of SPF in pharmaceutical dosage forms as per ICH guidelines [20]. The goal of this study is to develop rapid HPLC method for the analysis of SPF in bulk drug samples and tablet formulations using the most commonly employed column C18 with UV detection at suitable wavelength. MATERIALS AND METHODS Chemicals and reagents SPF active pharmaceutical ingredient (API) was obtained from FDC Limited, Mumbai, India. HPLC grade acetonitrile, water and analytical reagent (AR) grade Potassium dihydrogen orthophosphate dehydrate and orthophosphoric acid from Merk chemicals, Mumbai, India. While Commercial tablets of SPF was acquired from local market. Sparcip100mg (Cipla), Zospar 200mg (FDC Ltd.,), Mumbai, India are the commercial formulations. Instruments The HPLC analysis was performed on a high pressure isocratic high performance liquid chromatograph (SHIMADZU LC-20AT prominence liquid chromatograph) equipped with two LC-20AT VP pumps, manual injector with loop volume of 20 µL (Rheodyne), programmable variable wavelength SHIMADZU SPD-20A prominence UV-Vis detector and WELCHROM C18 Column with 250mm x 4.6mm i.d and 5µm particle size. The HPLC system was equipped with DzSpinchrom CFRdz software. Moreover an analytical balance (Shimadzhu TX223L), digital pH meter (systronics model 802), a sonicator (Spectra lab, model UCB 40), UV-Visible spectrophotometer (Systronics model-2203) were used in present study. Vol 7, Suppl 1, 2014 ISSN - 0974-2441