AN IMPROVED RP-HPLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF ASPIRIN, ATORVASTATIN, AND CLOPIDOGREL IN PHARMACEUTICAL FORMULATION USING EXPERIMENTAL DESIGN METHODOLOGY
Keywords:
Central composite design, Multiple response optimizations, HPLC method, Aspirin, Atrovastatin, and ClopidogrelAbstract
Objective: In this study an improved RP-HPLC method was developed for the simultaneous estimation of Aspirin, Atrovastatin, and Clopidogrel in pharmaceutical dosage form. For improving the separation, an experimental design approach was employed.
Methods: Factors–independent variables (Organic modifier, pH of the mobile phase and flow rate) were extracted from the preliminary study and as dependent three responses variables viz. Capacity factor of tR1, resolution between Atorvastatin and internal standard, retention time of tR4 were selected. To improve method development and optimization, Derringer's desirability function was applied to simultaneously optimize the chosen three responses.
Results: The procedure allowed deduction of optimal conditions and the predicted optimum was Acetonitrile: Methanol: 0.1% of Triethylamine (52:05:43, v/v/v), pH of the aqueous phase adjusted at to 3.0 with 10 % ortho phosphoric acid, and the separation was achieved within 8 minutes. The method showed good agreement between the experimental data and predictive value throughout the studied parameter space.
Conclusion: The optimized assay condition was validated according to International Conference on Harmonisation (ICH) guidelines to confirm specificity, linearity, accuracy and precision. The proposed validated method was successfully applied for the analysis of commercially available dosage form.
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References
Satoskar RS, Bhandarkar SD, Ainapure SS. Pharmacology and Pharmacotherapeutics. 23rd Ed. Popular Prakashan, Mumbai; 2013.
Getu K, Ann VS, Erwin A. Development and validation of a liquid chromatographic method for purity control of clopidogrel–acetylsalicylic acid in combined oral dosage forms. J Pharm Biomed Anal 2012;61:271-6.
Dipalipatel, Patel N, Reetavaishy. Development and Validation of RP-HPLC Method for Simultaneous Estimation of Aspirin and Esomeprazole Magnesium in Tablet Dosage Form. J Chem 2013.
Milkica Crevar Sakac, Zorica Vujic, Jasmina Brboric, Vesna Kuntic, Snezana Uskokovic-Markovic. An Improved HPLC method with the aid of a chemometric protocol: simultaneous determination of atorvastatin and its metabolites in plasma. Molecules 2013;18(3):2469-82.
Rumenka Petkovska. Experimental design approach for the development and validation of an enantiospecific RP-HPLC method for simultaneous determination of clopidogrel and related compounds. Maced J Chem Chem Eng 2008;27:1.
Getukahsay. Development and validation of a liquid chromatographic method for purity control of clopidogrel-acetylsalicylic acid in combined oral dosage forms. J Pharm Biomed Anal 2012;61:581-9.
Mahmoud Mohamed Issa. Resolution of ternary mixture of aspirin, atorvastatin, and clopidogrel by Chemometric-Assisted UV spectroscopic and liquid chromatography methods. Int J Spectr 2013.
Valliappan K, Sreejanardhanan V, Venkatesan P. Direct chiral HPLC method for the simultaneous determination of warfarin enantiomers and its impurities in raw material and pharmaceutical formulation: application of chemometric protocol. Chromatographia 2013;76:287-92.
Marcia CB, Isabel CSFJ, Roy EB. Combined column–mobile phase mixture statistical design optimization of high-performance liquid chromatographic analysis of multicomponent systems. J Chromatography A 2009;1216:1439-49.
Sivakumar T, Manavalan R, Muralidharan C, Valliappan K. Multi-criteria decision making approach and experimental design as chemometric tools to optimize HPLC separation of domperidone and pantoprazole. J Pharm Biomed Anal 2007;43:1842-8.
Kulikov AU, Zinchenko AA. Development and validation of reversed phase high performance liquid chromatography method for determination of dexpanthenol in pharmaceutical formulations. J Pharm Biomed Anal 2007;43:983-8.
Richard GB. Chemometrics Data Analysis for the Laboratory and Chemical Plant. 1st Ed. Wiley, England; 2003.
Sivakumar T, Manavalan R, Muralidharan C, Valliappan K. An improved HPLC method with the aid of a chemometric protocol: simultaneous analysis of amlodipine and atorvastatin in pharmaceutical formulations. J Sep Sci 2007;30(18):3143-53.
Beg QK, Sahai V, Gupta R. Statistical media optimization and alkaline protease production from Bacillus mojavensis in a bioreactor. Process Biochem 2003;39:203-9.
Biljana J, Mirjana M, Darko I, Andelija M. Experimental design in chromatographic analysisof pramipexole and its impurities. Acta Chem Slov 2007;54:49-54.
Derringer G, Suich R. Simultaneous optimization of several response variables. J Qual Technol 1980;12:214-9.
Wester P, Gottfries J, Johansson K, Klintebck F, Winblad B. Simultaneous liquid chromatographic determination of seventeen of the major monoamine neurotransmitters, precursors and metabolites: I. Optimization of the mobile phase using factorial designs and a computer program to predict chromatograms. J Chromatography B 1987;415:261-74.
Kleinschmidt G, Ermer J, Miller JHM. (Eds). Method Validation in Pharmaceutical Analysis. A Guide to best practice, Wiley-VCH, Weinheim; 2005. p. 195-226.