A VALIDATED STABILITY INDICATING RP-HPLC METHOD FOR THE SIMULTANEOUS DETERMINTION OF ATORVASTATIN CALCIUM AND EZETIMIBE HYDROCHLORIDE IN BULK AND TABLET DOSAGE FORM
Objective: To develop a simple, selective and precise stability indicating reverse-phase high performance liquid chromatography method for the simultaneous estimation of atorvastatin calcium and ezetimibe hydrochloride in bulk and tablet dosage form.
Methods: The chromatographic separation was performed by Agilent Zorbax column (250Ã—4.6 mm, 5 Âµm) using methanol: 0.1 % v/v orthophosphoric acid in water (65:35) as mobile phase at flow rate of 1.0 ml/min with injection volume 20 Âµl and the detection was carried out using UV detector at 240 nm. The method was validated as per ICH guidelines.
Results: The retention time for atorvastatin calcium (ATV) and ezetimibe hydrochloride (EZT) was found to be 6.81 min and 4.96 min respectively. The linear regression analysis data for the calibration plots showed good linear relationship in the concentration range of 5-50 Âµg/ml for both ATV and EZT. The percentage recoveries of ATV and EZT in the marketed dosage form were found to be 100.82 and 94.27 respectively. The correlation coefficients for ATV and EZT were 0.9983 and 0.998 respectively. The percentage degradation at different stress conditions like acid, alkaline, oxidative and photolytic for atorvastatin calcium were found to be 14.91, 8.26, 8.02 and 2.65 respectively and for ezetimibe hydrochloride, found to be 9.70, 32.18, 2.51 and 0.16 respectively.
Conclusion: The developed method was successfully validated as per ICH guidelines. This method is simple, selective, linear, precise, accurate and sensitive, and can be used for routine analysis of tablet dosage forms containing both the drugs.
Keywords: Atorvastatin calcium, Ezetimibe hydrochloride, RP-HPLC, Stress degradation
Clinical Pharmacology Online. Gold Standard Multimedia; 2005. Available From http://www.cp.gsm.com. [Last accessed on 25 Oct 2005].
Drug Facts and Comparisons. St. Louis, MO: Wolters Kluwer Health; 2005. p. 533-49.
Kosoglou T, Statkevich P, Johnson-Levonas AO, Paolini JF, Bergman AJ, Alton KB. Ezetimibe: a review of its metabolism, pharmacokinetics, and drug interactions. Clin Pharmacokinet 2005;44:467-94.
Lea AP, McTavish D. Atorvastatin: a review of its pharmacology and therapeutic potential in the management of hyperlipidemias. Drugs 1997;53:828-47.
Remington-The science and practice of pharmacy. Vol. II. 21st ed. In: Chapter 68: Anna MW. Cardiovascular drugs. New Delhi: Wolters Kluwer (India) Pvt. Ltd; 1995. p. 1368.
Eric B, Philippe G, Philippe T. Perspectives in cholesterol-lowering therapy. The role of ezetimibe, a new selective inhibitor of intestinal cholesterol absorption. Circulation 2003;107:3124-8.
Jamshidi A, Nateghi AR. HPTLC determination of atorvastatin in plasma. Chromatographia 2007;65:763-6.
Beata S, Lukasz K. Validation of HPLC method for the determination of atorvastatin in tablets and for monitoring stability in the solid phase. Acta Polonia Pharm Drug Res 2006;63:471-6.
Gholamreza B, Bahareh M, Shahla M, Amir K. Determination of atorvastatin in human plasma serum by reversed-phase high-performance liquid chromatography with UV detection. J Chromatogr B 2005;826:41-5.
Altuntas TG, Erk N. Liquid chromatographic determination of atorvastatin in bulk drug, tablets, and human plasma. J Liq Chromatogr Relat Technol 2004;27:83-93.
Sidika E, Esra SA, Lale E, Samiye F. An HPLC method for the determination of atorvastatin and its impurities in bulk drug and tablets. J Pharm Biomed Anal 2003;33:1017-23.
Saranjit S, Baljinder S, Rakesh B, Lalit W, Rahul S. Stress degradation studies on ezetimibe and development of a validated stability-indicating HPLC assay. J Pharm Biomed Anal 2006;41:1037-40.
Oleveira PR, Brum Junior L, Fronza M, Bernadi LS, Masiero SMK, Dalmora SL. Development and validation of a liquid chromatography-tandem mass spectrometry method for the determination of ezetimibe in human plasma and pharmaceutical formulations. Chromatographia 2006;63:315-20.
Shuijun Li, Gangyi Liu, Jingying Jia, Xiaochuan Li, Chen Yu. Liquid chromatography-negative ion electrospray tandem mass spectrometry method for the quantification of ezetimibe in human plasma. J Pharm Biomed Anal 2006;40:987-92.
Stefan O, Eberhard S, Ingolf C, Werner S. A LC-MS/MS method to quantify the novel cholesterol-lowering drug ezetimibe in human serum, urine, and feces in healthy subjects genotyped for SLCO1B1. J Chromatogr B 2006;830:143-50.
Sistla R, Tata VSSK, Kashyap YV, Chandrasekar D, Diwan PV. Development and validation of a reversed-phase HPLC method for the determination of ezetimibe in pharmaceutical dosage forms. J Pharm Biomed Anal 2005;39:517-22.
Dixit RP, Barhate CR, Nagarsenker MS. Stability-indicating HPTLC method for simultaneous determination of ezetimibe and simvastatin. Chromatographia 2008;67:101-7.
Petkovska R, Cornett C, Dimitrovska A. Development and validation of rapid resolution RP-HPLC method for simultaneous determination of atorvastatin and related compounds by use of chemometrics. Anal Lett 2008;41:992-1009.
Jain N, Jain R, Swami H, Jain DK. RP-HPLC method for simultaneous estimation of simvastatin and ezetimibe in bulk drug and its combined dosage form. Asian J Res Chem 2008;1:29-31.
Lucie N, Dalibor S, Petr Solich. HPLC methods for the determination of simvastatin and atorvastatin. TrAC Trends Anal Chem 2008;27:352-67.
Chaudhari BG, Patel NM, Shah PB. Stability indicating RP-HPLC method for simultaneous determination of atorvastatin and amlodipine from their combination drug products. Chem Pharm Bull 2007;55:241-6.
Mohammadi A, Ansari DM, Ghorbani BF, Hashem M, Walker RB. A stability-indicating high-performance liquid chromatography (HPLC) assay for the simultaneous determination of atorvastatin and amlodipine in commercial tablets. J Chromatogr B: Anal Technol Biomed Life Sci 2007;846:215-21.
Ozaltin N, Ucakturk E. Simultaneous determination of ezetimibe and simvastatin in pharmaceutical formulations by dual-method gradient LC. Chromatographia 2007;66:87-91.
Chaudhari BG, Patel NM, Shah PB. Stability-indicating reversed-phase liquid chromatographic method for simultaneous determination of simvastatin and ezetimibe from their combination drug products. J AOAC Int 2007;90:1242-9.
Oliveira PR, Barth T, Todeschini V, Dalmora SL. Simultaneous liquid chromatographic determination of ezetimibe and simvastatin in pharmaceutical products. J AOAC Int 2007;90:1566-72.
Borek-Dohalsky V, Huclova J, Barret B, Nemec B, Ulc I, Jelinek I. Validated HPLC-MS-MS method for simultaneous determination of atorvastatin and 2-hydroyatorvastatin in the human plasma-pharmacokinetic study. Anal Bioanal Chem 2006;386:275-85.
AlShehri MM. A validated capillary electrophoresis method for simultaneous determination of ezetimibe and atorvastatin in pharmaceutical formulations. Saudi Pharm J 2012;20:143-8.
ICH Q1AR, Stability Testing of New Drug Substances and Products, International Conference on Harmonization IFPMA, Geneva, Switzerland; 2000.
U. S. Pharmacopeia. 24th ed. U. S. Pharmacopeial Convention, Inc., Rockville, MD; 2000.
Ermer J, Miller JH. Method validation in pharmaceutical analysis. 1st Ed. Wiley-VCH Publishers, Weinheim, Germany; 2005.
Swartz ME, Krull IS. Analytical method development and validation. Marcel Dekker, New York, NY; 1997.
ICH Q2A, Validation of Analytical Procedures, Methodology. International Conference on Harmonization, Brussels, Belgium; 1995.
Riley CM, Rosanske TW. Development and validation of analytical methods. Elsevier, Amsterdam, The Netherlands; 1996.
Raul SK, Aravelli AB, Jhansi D. RP-HPLC method development and validation for the simultaneous estimation of atorvastatin and ezetimibe in pharmaceutical dosage form. Asian J Pharm Clin Res 2015;8:178-81.
Kumar P, Ahmed Y. Stability is indicating method development for simultaneous determination of ezetimibe and atorvastatin in dosage form by RP-HPLC. Der Pharm Sinica 2012;3:413-21.
Kumar SA, Debnath M, Seshagiri Rao JVLN, Sankar DG. Stability indicating RP-HPLC analytical method development and validation for simultaneous estimation of atorvastatin and ezetimibe in bulk as well in pharmaceutical dosage form by using PDA detector. Der Pharm Lett 2014;6:37-55.