title: A UHPLC-MS/MS method for the quantification of direct antiviral agents simeprevir, daclatasvir, ledipasvir, sofosbuvir/GS-331007, dasabuvir, ombitasvir and paritaprevir, together with ritonavir, in human plasma.
abstract: To date, the new standard for treatment of chronic hepatitis C is based on the administration of novel direct acting antivirals. Among these, sofosbuvir, simeprevir, daclatasvir, ledipasvir, dasabuvir, ombitasvir and paritaprevir already entered the clinical use. Anyway, since few pharmacokinetic studies have been conducted on these drugs in a "real life" context poor knowledge is available about their optimal therapeutic range. Without this background, therapeutic drug monitoring is not applicable for treatment optimization. Up to now, a few methods are reported to quantify these drugs in human plasma, and none of them in a simultaneous way. The aim of this work was to develop and validate a simple, fast and cheap, but still reliable UHPLC-MS/MS method for the quantification of these drugs, feasible for a clinical routine use. Solid phase extraction was performed using HLB C18 96-well plates. Chromatographic separation was performed on a BEH C18 1.7μm, 2.1mm×50mm column, settled at 50°C, with a gradient run of two mobile phases: ammonium acetate 5mM (pH 9.5) and acetonitrile, with a flow rate of 0.4mL/min for 5min. Tandem-mass detection was carried out in positive electrospray ionization mode. Both inter and intraday imprecision and inaccuracy were below 15%, as required by FDA guidelines, while both recoveries and matrix effects resulted within the acceptance criteria. The method was tested on 80 patients samples with good performance. Being robust, simple and fast and requiring a low plasma volume, this method resulted eligible for a clinical routine use.
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Ariaudo
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Favata
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy. Electronic address: amedeo.denicolo@unito.it.
authors.forename: Amedeo
authors.initials: A
authors.lastname: De Nicolò
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy; "CoQuaLab", Academic spin-off of University of Turin, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Simiele
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Paglietti
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors.forename: Lucio
authors.initials: L
authors.lastname: Boglione
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors.forename: Chiara Simona
authors.initials: CS
authors.lastname: Cardellino
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Carcieri
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Di Perri
authors.affiliation: Unit of Infectious Diseases, University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: D'Avolio
mesh_terms: D000998:Antiviral Agents / Q000097:blood*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000998:Antiviral Agents
keywords: DAAs; HCV; Liquid chromatography; Multiplexed; TDM; Tandem mass spectrometry