Novel determination of sofosbuvir and velpatasvir in human plasma by UPLC–MS/MS method: Application to a bioequivalence study

Abstract

A novel and sensitive LC-MS/MS method was developed, optimized and validated for quantification of sofosbuvir (SOF) and velpatasvir (VEL) in human plasma using ledipasvir as an internal standard (IS). Sample preparation was done using acetonitrile for precipitation of plasma proteins. Chromatographic analysis was done on an Acquity UPLC BEH C column using 0.1% formic acid and acetonitrile as a mobile phase. The Xevo TQD LC-MS/MS system was run with electrospray ionization mode. The developed method was optimized and then validated according to the US Food and Drug Administration guidelines. Linearity was found to be in the range of 0.25-3500 ng/mL for SOF and 1-1000 ng/mL for VEL. A short run time of 1.5 min allows swift analysis of many plasma samples per day. The developed method was successfully utilized for estimating both SOF and VEL in the plasma of healthy human volunteers participated in a bioequivalence study.

Recent advances in the chromatographic determination of the most commonly used anti‐hepatitis C drug sofosbuvir and its co‐administered drugs in human plasma

“…A mobile phase composed of ACN and FA was frequently used in most of the reported UPLC methods for the analysis of sofosbuvir due to its superior separation characteristics (Notari et al, 2018; Pan et al, 2016; Rezk et al, 2015; Rezk et al, 2018; Rezk, Basalious, & Amin, 2016; Rezk, Bendas, et al, 2016; Semreen et al, 2019; Shi et al, 2015). This mixture eluted the analytes and IS in a narrow range, which in turn reflects on the compensation for matrix effects (Rezk et al, 2015; Rezk, Basalious, & Amin, 2016) and the fluctuation in ionization of MS analysis, with a reduction in the total time needed for the analysis (Rezk et al, 2018).…”

“…The use of isocratic elution allows omitting the equilibration step after finishing gradient (Rezk et al, 2018), and the use of gradient elution was found to compensate for the carry‐over problems in the simultaneous determination of sofosbuvir, GS‐331007, grazoprevir, ledipasvir, simeprevir, daclatasvir, elbasvir, and velpatasvir (van Seyen et al, 2019).…”

“…The use of ketotifen and ledipasvir as IS was also exploited owing to their stability and compatibility with the response of the detector; in addition to their similar physicochemical properties to sofosbuvir, similar extraction procedure and retention time were required (Rezk et al, 2018; Semreen et al, 2019). Famotidine (Rezk et al, 2015), midazolam (Shi et al, 2015), eplerenone (Rezk, Basalious, & Amin, 2016; Rezk, Bendas, et al, 2016), 6,7‐dimethyl‐2,3‐di(2‐pyridyl), and quinoxaline (Ariaudo et al, 2016; Notari et al, 2018) were also employed as ISs for the UPLC analysis of sofosbuvir.…”

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“…The literature review discloses that a very few spectroscopic, [7,8] LC-MS/MS, [9] and high performance liquid chromatographic techniques [10][11][12][13][14][15][16] have been reported for the estimation of VXR, SFR, and VLR. Based on the reported HPLC methods, there is a need to develop a rapid, sensitive reversed-phase-UPLC method for simultaneous estimation of VXR, SFR, and VLR in bulk and formulations.…”

Stability Indicating HPTLC Method for Sofosbuvir and Velpatasvir in Combination

“…After a thorough literature survey, it is observed that there are many analytical methods reported for analysis of SOF and VEL, either single or in combination, but no SIM-HPTLC method was found for combination. The reported methods of SOF combination with VEL include analytical methods like LC-MS/MS, [1] HPLC, [2][3][4][5] UPLC MS/MS [6] and UPLC. [7]…”