title: Development of a simple HPLC-UV method for the determination of the hepatitis C virus inhibitor simeprevir in human plasma.
abstract: A simple high-performance liquid chromatography method for the determination of the hepatitis C virus protease inhibitor simeprevir in human plasma was developed and validated. The method involved a rapid and simple solid-phase extraction of simeprevir using Oasis HLB 1cc cartridges, isocratic reversed-phase liquid chromatography on an XTerra RP18 (150 mm×4.6 mm, 3.5 μm) column, and ultraviolet detection at 225 nm. The mobile phase consisted of phosphate buffer (pH 6, 52.5 mM) and acetonitrile (30:70, v/v). This assay proved to be sensitive (lower limit of quantification of 0.05 μg/mL), linear (correlation coefficients ≥0.99), specific (no interference with various potentially co-administrated drugs), reproducible (both intra-day and inter-day coefficients of variation ≤8.3%), and accurate (deviations ranged from -8.0 to 1.2% and from -3.3 to 6.0% for intra-day and inter-day analysis, respectively). The method was applied to therapeutic monitoring of patients undergoing simeprevir treatment for hepatitis C and proved to be robust and reliable. Thus, this method provides a simple, sensitive, precise and reproducible assay for dosing simeprevir that can be readily adaptable to routine use by clinical laboratories with standard equipment.
authors.affiliation: Department of Molecular Medicine, University of Padua, Padua, Italy.
authors.forename: Giulio
authors.initials: G
authors.lastname: Nannetti
authors.affiliation: Department of Molecular Medicine, University of Padua, Padua, Italy; Clinical Microbiology and Virology Unit, Padua University Hospital, Padua, Italy.
authors.forename: Silvana
authors.initials: S
authors.lastname: Pagni
authors.affiliation: Department of Molecular Medicine, University of Padua, Padua, Italy; Clinical Microbiology and Virology Unit, Padua University Hospital, Padua, Italy.
authors.forename: Saverio G
authors.initials: SG
authors.lastname: Parisi
authors.affiliation: Department of Molecular Medicine, University of Padua, Padua, Italy.
authors.forename: Alfredo
authors.initials: A
authors.lastname: Alberti
authors.affiliation: Department of Molecular Medicine, University of Padua, Padua, Italy; Clinical Microbiology and Virology Unit, Padua University Hospital, Padua, Italy. Electronic address: arianna.loregian@unipd.it.
authors.forename: Arianna
authors.initials: A
authors.lastname: Loregian
authors.affiliation: Department of Molecular Medicine, University of Padua, Padua, Italy; Clinical Microbiology and Virology Unit, Padua University Hospital, Padua, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Palù
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D016174:Hepacivirus / Q000187:drug effects; D006801:Humans; D010949:Plasma / Q000737:chemistry*; D015203:Reproducibility of Results; D000069616:Simeprevir / Q000097:blood* / Q000737:chemistry* / Q000494:pharmacology; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
chemical_list: D000069616:Simeprevir
keywords: HCV protease inhibitor; HPLC–UV; Simeprevir; Solid-phase extraction; TMC435; Therapeutic drug monitoring