A VALIDATED LC-MS/MS METHOD FOR THE ESTIMATION OF BOCEPREVIR AND BOCEPREVIR D6 (IS) IN HUMAN PLASMA EMPLOYING LIQUID-LIQUID EXTRACTION
Journal Title: International Journal of Pharmacy and Pharmaceutical Sciences - Year 2016, Vol 8, Issue 7
Abstract
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
Authors and Affiliations
Bhetanabotla Chandramowli, Bigala B. Rajkamal
Keywords
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