A novel, rapid and simple UPLC–MS/MS method for quantification of favipiravir in human plasma: Application to a bioequivalence study
Mamdouh R. Rezk, Kamal A. Badr, Naglaa S. Abdel‐Naby, Magy M. Ayyad
Abstract
Abstract A novel, simple and rapid UPLC–MS/MS method was developed and validated for determination of favipiravir (FAV) in human plasma. Lamivudine was used as an internal standard (IS). The Xevo TQD LC–MS/MS was operated under the multiple‐reaction monitoring mode using electrospray ionization. Precipitation with acetonitrile was used in sample preparation as it gives relatively cleaner plasma samples. The prepared samples were chromatographed using an Acquity UPLC ® HSS C 18 (100 × 2.1 mm, 1.8 μm) column. The mobile phase was composed of ammonium formate and methanol in a gradient mode that was pumped at a flow rate of 0.35 ml/min. The developed method was validated as per the FDA guidelines and linearity was in the range of 0.25–16 μg/ml for FAV. The intra‐ and inter‐day precision and accuracy results were within the acceptable limits. A run time of 4.5 min and a low quantification limit of FAV allowed the application of the developed method for the determination of FAV in a bioequivalence study in healthy human volunteers.