Novel and sensitive UPLC–MS/MS method for quantification of sofosbuvir in human plasma: application to a bioequivalence study |
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| Authors: | Mamdouh R Rezk Emad B Basalious Mohammed E Amin |
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| Affiliation: | 1. Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt;2. Pharmaceutics and Industrial Pharmacy Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt;3. Advanced Research Center, Nasr City, Cairo, Egypt |
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| Abstract: | A novel and sensitive LC–MS/MS method was developed and validated for determination of sofosbuvir (SF) using eplerenone as an internal standard. The Xevo TQD LC–MS/MS was operated under the multiple‐reaction monitoring mode using electrospray ionization. Extraction with tert‐butyl methyl ether was used in sample preparation. The prepared samples were chromatographed on Acquity UPLC BEH C18 (50 × 2.1 mm, 1.7 μm) column by pumping 0.1% formic acid and acetonitrile in an isocratic mode at a flow rate of 0.35 mL/min. Method validation was performed as per the US Food and Drug Administration guidelines and the standard curves were found to be linear in the range of 0.25–3500 ng/mL for SF. The intra‐ and inter‐day precision and accuracy results were within the acceptable limits. A very short run time of 1 min made it possible to analyze more than 500 human plasma samples per day. A very low quantification limit of SF allowed the applicability of the developed method for determination of SF in a bioequivalence study in human volunteers. Copyright © 2016 John Wiley & Sons, Ltd. |
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| Keywords: | sofosbuvir UPLC‐MS/MS plasma validation bioequivalence |
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