ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE DETERMINATION OF HYDROCHLOROTHIAZIDE, AMLODIPINE BESYLATE AND TELMISARTAN HYDROCHLORIDE IN MULTICOMPONENT TABLET DOSAGE FORM AND IN BIORELEVANT MEDIA (FASSIF) BY RP-HPLC TECHNIQUES Original Article MADHUKAR A a,b *, N. KANNAPPAN b , MAHENDRA KUMAR CB * c a Bright Labs, Kothapet, Dilshuknagar, Hyderabad, Telangana, INDIA, b Department of Pharmacy, Annamalai University, Annamalai Nagar, Chidambaram, Tamil Nadu, INDIA, c Received: 14 Oct 2014 Revised and Accepted: 15 Nov 2014 Department of Pharmacy, St. Mary’s College of Pharmacy, Secunderabad, Telangana, INDIA. Email: dr.amk2014@gmail.com ABSTRACT Objective: A simple, rapid, and precise reverse phase high performance liquid chromatographic (RP-HPLC) method for simultaneous analysis of Hydrochlorothiazide (HTZ), Amlodipine besylate (AML) and Telmisartan hydrochloride (TLM) in a tablet dosage form and in Biorelevant media has been developed and validated. Methods: This method was performed with a thermosil C 18 Results: By the method HTZ, AML and TLM were eluted with retention times of 1.823, 2.639, and 4.198 min, respectively. The method was continued and validated accordance with ICH guidelines. Validation revealed the method is rapid, specific, accurate, precise, reliable, and reproducible. Calibration curve plots were linear over the concentration ranges 6.25-100μg/mL for HTZ, 2.5-40μg/mL for AML, and 20-320μg/mL for TLM. Limits of detection (LOD) were 0.004, 0.0016, and 0.0128μg/ml and limits of quantification (LOQ) were 0.013, 0.0052, and 0.0416μg/mL for HTZ, AML and TLM respectively. (4.6 × 100 mm i. d., 3.7 μm particle column with 40:60 (v/v) 20 mM potassium dihydrogen orthophosphate buffer: methanol as mobile phase at a flow rate of 1.0 ml/min. UV detection at 248 nm. Conclusion: The statistical analysis was proves the method is suitable for the analysis of HTZ, AML and TLM as a bulk, in tablet dosage form and in biorelevant media without any interference from the excipients. It was also proved study for degradation kinetics of three drugs. It may be extended for its estimation in plasma and other biological fluids. Keywords: Hydrochlorothiazide (HTZ), Amlodipine besylate (AML) and Telmisartan hydrochloride (TLM), RP-HPLC, Validation, FaSSIF. INTRODUCTION Hydrochlorothiazide (HTZ) chemically 6-chloro-1,1-dioxo-3,4- dihydro-2H-1,2,4-benzothiadiazine-7-sulfonamide (Fig. 1). It is a diuretic drug and derivative of the thiazide class [1]. Amlodipine (AML) chemically (RS)-3-ethyl-5-methyle-2-[(2-aminoethoxy) methyl]-4-(2-chlorophenyl)-6-methyl-1,4-dihydropyridine-3,5-di carboxylate (Fig. 1). It is a long-acting dihydropyridine-type (DHP) of calcium channel blocker [2]. Amlodipine is also acts as a functional inhibitor of acid sphingomyelinase [3]. Telmisartan (TLM) chemically 2-(4-{[4-methyl-6-(1-methyl-1H-1,3-benzodiazol-2-yle)- 2-propyle-1H-1,3-benzodiazol-1-yle]methyl}phenyl)benzoic acid (Fig. 1). It is an angiotensin II receptor blocking agent it shows high affinity to the angiotensin II receptor type-1 (AT1) [4]. In the scientific literature, analysis of HTZ, AML, and TLM has been reported as individual ingredients and in combination with other compounds. Analytical methods have included estimation of HTZ [5], AML [6], TLM [7, 8] individually. And in two component formulations of HTZ and AML have been analyzed in combination [9, 10]. Simultaneous HPLC analysis of HTZ and TLM [11] and in combinations of HTZ, AML and TLM analyzed in HPLC [12-14] and with other drugs had also been reported [15-17]. No other chromatographic methods are found for simultaneous analysis of HTZ, AML, and TLM in a combined dosage form and in biorelevant media. The method described is rapid, economical, precise, and accurate and can be used for routine analysis of tablets. It was validated as per ICH guidelines [18-20]. HTZ AML TLM Fig. 1: Chemical structures of Hydrochlorothiazide (HTZ), Amlodipine besylate (AML) and Telmisartan (TLM) MATERIALS AND METHODS Experimental Materials and methods Pharmaceutical grade working standards Hydrochlorothiazide (HTZ), Amlodipine besylate (AML), Telmisartan hydrochloride (TLM) were obtained from Hetero Labs, Jedcharla, India. All chemicals and reagents were HPLC grade and were purchased from Merck Chemicals, Mumbai, India. Instrumentation The analysis was performed using Waters-2695 (Modal Alliance) High Performance liquid chromatography, analytical balance International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 7, Issue 1, 2015 Innovare Academic Sciences