INTERNATIONAL JOURNAL OF PHARMACEUTICS & DRUG ANALYSIS VOL.4 ISSUE 2, 2016; 64 – 71 ; http://ijpda.com ; ISSN: 2348-8948 64 RESEARCH ARTICLE DEVELOPMENT, VALIDA- TION & STRESS DEGRA- DATION STUDIES OF DARUNAVIR BY REVERSE PHASE-HIGH PERFOR- MANCE LIQUID CHROMATOGRAPHGY (RP-HPLC) PVSR Mohana Rao 1 , Dr. Jagannadha Rao Vepa 2 , Tata Santosh 3 , Dr. K Raghu Babu 1 * 1 Andhra University College of Engineering, Andhra Universtiy, Andhra Pradesh, India. 2 Anil Neerukonda Institute of Technology & Science, Vi- sakhapatnam, Andhra Pradesh, India. 3 Corpuscle Research Solutions, Andhra Pradesh, India. Date Received: 30 th January 2016; Date Accepted: 11 th February 2016 Date published: 12 th February 2016 Email: drraghualways@yahoo.co.in Abstract: A simple, selective, rapid, precise and economi- cal reverse phase high pressure liquid chromatographic method has been developed for the estimation of Darunavir in pharmaceutical dosage form. The mobile phase consisted of 80:20% (v/v) of Methanol & 0.1% or- tho-phosphoric acid was used and operated on isocratic mode. The flow rate is 1.0 mL/min. Chromatographic de- termination of Darunavir was performed on Agilent Pola- ris C18 column (150 X 4.6 mm id, ODS 2,5µ m). The wave- length of detection is 265nm. The injection volume is 20µL. The retention time of Darunavir is 2.42 ± 0.01minutes. The developed method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantification and solution stability. The influence of Acid, Alkaline, Oxidative stress, Photolytic stress con- dition on Darunavir was studied. Results indicated com- pleted degradation in Alkaline medium. The proposed method has been successfully used for the estimation in tablet dosage forms. Key words: Darunavir; HPLC, Tablet formulation. Introduction Darunavir ethanolate (DRV) is an oral anti-retroviral agent which selectively inhibits the cleavage of Human immunodeficiency virus (HIV-1) encoded Gas- polyproteins in infected cell, thereby preventing the for- mation of mature virus. Darunavir ethanolate is chemical- ly [(1S,2R)-3-[[(4-amino phenol) sulfonyl](2-methyl pro- pyl)amino]-2-hydroxy-1-(phenylmethyl)propyl]-carbamic acid (3R,3aS,6aR)-hexahydrofuro[2,3-b]furan-3-yl ester monoethanolate [1] (fig. 1). Darunavir was designed to form robust interactions with the protease enzyme from many strains of HIV, including strains from the treatment- experienced patients with multiple resistance mutations to Protease inhibitors.[2] It blocks HIV protease, an en- zyme which is needed for HIV to multiply. According to in vitro experiments, DRV was active against HIV-1 with PI-resistance mutations and against PI-resistance clinical isolates [3-4]. This drug is effective in patients experi- enced in anti-retroviral treatment, such as those carrying HIV-1 strains which are resistance to more than one PI [5]. The use of advanced instrumentation techniques for the analysis of drugs has been discussed elsewhere [6]. Litera- ture survey revealed that different analytical methods have been reported for the determination of DRV in plasma using liquid chromatography coupled with tan- dem mass spectroscopy [7]; simultaneous determination of DRV with other anti-retroviral agents in plasma[8-9]. Few HPTLC method for determination of darunavir in rat plasma and in tablet dosage form its application to phar- macokinetics studies [10]. Infrared Spectroscopy method for determination of Darunavir in tablets [11]. Few meth- ods had been developed for determination of Darunavir by HPLC [12-16] and Spectrophotometric method [17]; and electrophoretic method for the separation of DRV[18]. Reported HPLC methods require more time for sample analysis resulting in lesser throughput. Therefore, the present work involves the development of a rapid RP- HPLC method for estimation of Darunavir ethanolate in bulk and tablet dosage form. Validation as per USFDA & ICH guidelines [19], [20] is done along with stress degra- dation study. The aim of the present study is to develop a simple, precise and accurate reversed-phase HPLC meth- od for the development and validation of Darunavir in bulk drug samples and in pharmaceutical dosage form.