Hindawi Publishing Corporation Chromatography Research International Volume 2012, Article ID 870951, 9 pages doi:10.1155/2012/870951 Research Article A Stability Indicating UPLC Method for the Determination of Tramadol Hydrochloride: Application to Pharmaceutical Analysis Kanakapura B. Vinay, Hosakere D. Revanasiddappa, Cijo M. Xavier, Pavagada J. Ramesh, and Madihalli S. Raghu Department of Chemistry, University of Mysore, Manasagangothri, Karnataka, Mysore 570 006, India Correspondence should be addressed to Hosakere D. Revanasiddappa, hdrevanasiddappa@yahoo.com Received 28 February 2012; Accepted 28 March 2012 Academic Editor: Antonio Mart´ ın-Esteban Copyright © 2012 Kanakapura B. Vinay et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The use of Ultra Performance Liquid Chromatography (UPLC), with a rapid 5-minute reversed phase isocratic separation on a 1.7 μm reversed-phase packing material to provide rapid “high throughput” support for tramadol hydrochloride (TMH) is demonstrated. A simple, precise and accurate stability-indicating isocratic UPLC method was developed for the determination of TMH in bulk drug and in its tablets. The method was developed using Waters Aquity BEH C18 column (100mm × 2.1 mm, 1.7 μm) with mobile phase consisting of a mixture of potassium dihydrogen phosphate buffer of pH 2.8 and an equal volume of acetonitrile (60 : 40 v/v). The eluted compound was detected at 226 nm with a UV detector. The standard curve of mean peak area versus concentration showed an excellent linearity over a concentration range 0.5–300 μg mL −1 TMH with regression coefficient (r) value of 0.9999. The limit of detection (S/N = 3) was 0.08 μg mL −1 and the limit of quantification (S/N = 10) was 0.2 μg mL −1 . Forced degradation of the bulk sample was conducted an accordance with the ICH guidelines. Acidic, basic, hydrolytic, oxidative, thermal and photolytic degradation were used to assess the stability indicating power of the method. TMH was found to degrade significantly in acidic, basic and oxidative stress conditions and stable in thermal, hydrolytic and photolytic conditions. 1. Introduction Tramadol hydrochloride (TMH), chemically known as (1R,2R)-rel-2-[(dimethylamino)methyl]-1-(3-methoxyphe- nyl)cyclohexanol (Figure 1), is a synthetic analogue of codeine and is a centrally acting analgesic agent [1]. It is metabolized by the cytochrome P450 enzyme system in the liver to form eleven metabolites of which o- desmethyltramadol (M1) predominates and has analgesic properties [2]. It has been used since 1977 for the relief of severe physical pain and has been the most widely sold opioid analgesic drug in the world [3]. TMH is official in European Pharmacopeia (EP) [4] which describe nonaqueous titration with perchloric acid as titrant the end point being located potentiometrically. Ultraviolet spectrophotometry [5, 6], high-performance liquid chromatography [6–9], thin layer chromatography-densitometry [10], capillary isotachophoresis [11], flow injection chemiluminescence spectrometry [12], voltametry [13–15], ion-selective-based potentiometry [16–22], visible spectrophotometry [23– 28], and titrimetry [26, 29] for determining TMH in pharmaceutical dosage forms. In addition, there have been reports of its assay when present in combination with other drugs. TDH and ibupro- fen were assayed simultaneously by first-order derivative spectrophotometry and HPLC with UV-detection [30]. TMH along with dexketoprofen and haloperidol have been determined by HPLC-diode array detection (DAD) method [31]. Simultaneous analysis of TMH and aceclofenac in a commercial tablet was accomplished also by HPLC with UV-detection method [32]. Simultaneous determination of paracetamol, TMH, and domperidone in combined dosage