A VALIDATED LC-MS/MS METHOD FOR THE ESTIMATION OF BOCEPREVIR AND BOCEPREVIR D6 (IS) IN HUMAN PLASMA EMPLOYING LIQUID-LIQUID EXTRACTION
Keywords:
Boceprevir, LC-MSMS, Human plasma, Liquid-Liquid ExtractionAbstract
Objective: This study was aimed to develop a simple, rapid, specific and precise liquid chromatography-tandem mass spectrophotometric (LC–MS/MS) validated method for quantification of Boceprevir and internal standard (ISTD) Boceprevir D6 in human plasma.
Methods: Plasma samples were pretreated with 100 µl of 0.1N Sodium Hydroxide and are subjected for Liquid-Liquid Extraction (LLE) using 2.5 ml of ethyl acetate. Chromatographic separation was achieved on Chromolith RP18e column (100 mmx4.6 mmx5 µm) with Acetonitrile: 20 mM Ammonium formate (80:20%v/v) as an isocratic mobile phase with a flow rate of 1.2 ml/min. the LC eluent was split, and approximately 0.1 ml/min was introduced into Tandem mass spectrometer using turbo Ion Spray interface at 400 °C. Quantitation was performed by transitions of m/z 586.2 precursor ion to the m/z 422.2 for Boceprevir and m/z 592.2/574.20 for Boceprevir D6.
Results: The concentrations of nine working standards showed linearity between 2 to 1000 ng/ml (r2 ≥ 0.998). Chromatographic separation was achieved within 3 min. The limit of Quantification (LOQ) was found to be 2ng/ml. The intraday and interday precision with quality control samples was found to be 0.09 to 3.17%.
Conclusion: The assay is suitable for pharmacokinetic study samples as demonstrated by its specificity, precision, accuracy, recovery, and stability characteristics.
Keywords: Boceprevir, LC-MS/MS, Human plasma, Liquid-Liquid Extraction
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References
Degertekin B, Lok AS. Update on viraral hepatitis. Curr Opin Gastroenterol 2007;41:306-11.
Njoroge FG, Chen KX, Shih NY, Piwinski JJ. Challenges in modern drug discovery: a case study of boceprevir, an HCV protease inhibitor for the treatment of hepatitis C virus infection. Acc Chem Res 2008;41:50–9.
http://www.drugbank.ca/drugs/DB08873. [Last accessed on 21 Jan 2016].
Shiny G, Satyavathi D. Development, and validation of rp-hplc method for the analysis of boceprevir and related impurities in bulk and pharmaceutical dosage forms. Indo Am J Pharm Res 2015;5:2391-401.
Ghosal A, Yuan Yuan, Wei Tong, Ai-Duen Su, Chunyan Gu, K Swapan Chowdhury et al. Characterization of human liver enzymes involved in the biotransformation of boceprevir, a hepatitis c virus protease inhibitor. Drug Metab Dispos 2011;39:213-22.
Xiaoyan Chu, Xiaoxin Cai, Donghui Cui, Cuyue Tang, Anima Ghosal, Grace Chan, et al. In vitro assessment of drug-drug interaction potential of boceprevir associated with drug metabolizing enzymes and transporters. Drug Metab Dispos 2013;41:668-81.
Aouri M, Moradpour D, Cavassini M, Mercier T, Buclin T, Csajka C, et al. Multiplex liquid chromatography-tandem mass spectrometry assay for simultaneous therapeutic drug monitoring of ribavirin, boceprevir, and telaprevir. Antimicrob Agents Chemother 2013;57:3147-58.
Farnika H, El Duweika J, Welschb C, Sarrazinb C, Lotscha J, Zeuzemb S, et al. Highly sensitive determination of HCV protease inhibitors boceprevir (SCH 503034) and telaprevir (VX 950) in human plasma by LC–MS/MS. J Chromatogr B 2009;31:4001-6.