title: UPLC-MS/MS method for the simultaneous quantification of three new antiretroviral drugs, dolutegravir, elvitegravir and rilpivirine, and other thirteen antiretroviral agents plus cobicistat and ritonavir boosters in human plasma.
abstract: Rilpivirine (RPV), dolutegravir (DTG) and elvitegravir (EVG) are the latest antiretroviral drugs approved for treatment of HIV infection. Currently, poor information is currently available concerning their pharmacokinetic and pharmacodynamic properties, thus making the use of therapeutic drug monitoring for these drugs not useful. This lack of information is partially due to the absence of an high-throughput method for their simultaneous quantification together with other antiretroviral drugs. In this work, we describe the development and validation of a new UPLC-MS/MS method to quantify these drugs, together with other fourteen antiretroviral agents, in human plasma. One hundred microliters of plasma samples were added with internal standard (6,7-Dimethyl- 2,3-di(2-pyridyl) quinoxaline), underwent a simple protein precipitation with methanol:acetonitrile (50:50v/v) followed by sample dilution with water. Chromatographic separation was performed on a Acquity®UPLC HSS T3 column (150mm x 2.1mm I.D) with a particle size of 1.8μm and compounds were detected with a tandem mass detector, monitoring two ion transitions for each drugs. The mean recovery of RPV, DTG and EVG were 101%, 87% and 112.3% respectively. Accuracy and precision inter/intra-day were below 15% for all drugs, in accordance to Food and Drug Administration guidelines requirements. The UPLC-MS/MS method reported here could be used routinely to monitor plasma concentrations of antiviral drugs, including RPV, DTG and EVG.
authors.affiliation: University of Turin, Department of Medical Sciences,(2) Laboratory of Clinical Pharmacology and Pharmacogenetics,(3) Amedeo di Savoia Hospital, CorsoSvizzera 164, 10149, Turin, Italy; "CoQuaLab", Academic Apin-off of University of Turin, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Simiele
authors.affiliation: University of Turin, Department of Medical Sciences,(2) Laboratory of Clinical Pharmacology and Pharmacogenetics,(3) Amedeo di Savoia Hospital, CorsoSvizzera 164, 10149, Turin, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Ariaudo
authors.affiliation: University of Turin, Department of Medical Sciences,(2) Laboratory of Clinical Pharmacology and Pharmacogenetics,(3) Amedeo di Savoia Hospital, CorsoSvizzera 164, 10149, Turin, Italy. Electronic address: amedeo.denicolo@unito.it.
authors.forename: Amedeo
authors.initials: A
authors.lastname: De Nicolò
authors.affiliation: University of Turin, Department of Medical Sciences,(2) Laboratory of Clinical Pharmacology and Pharmacogenetics,(3) Amedeo di Savoia Hospital, CorsoSvizzera 164, 10149, Turin, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Favata
authors.affiliation: University of Turin, Department of Medical Sciences,(2) Laboratory of Clinical Pharmacology and Pharmacogenetics,(3) Amedeo di Savoia Hospital, CorsoSvizzera 164, 10149, Turin, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Ferrante
authors.affiliation: University of Turin, Department of Medical Sciences,(2) Laboratory of Clinical Pharmacology and Pharmacogenetics,(3) Amedeo di Savoia Hospital, CorsoSvizzera 164, 10149, Turin, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Carcieri
authors.affiliation: University of Turin, Department of Medical Sciences,(2) Laboratory of Clinical Pharmacology and Pharmacogenetics,(3) Amedeo di Savoia Hospital, CorsoSvizzera 164, 10149, Turin, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Bonora
authors.affiliation: University of Turin, Department of Medical Sciences,(2) Laboratory of Clinical Pharmacology and Pharmacogenetics,(3) Amedeo di Savoia Hospital, CorsoSvizzera 164, 10149, Turin, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Di Perri
authors.affiliation: University of Turin, Department of Medical Sciences,(2) Laboratory of Clinical Pharmacology and Pharmacogenetics,(3) Amedeo di Savoia Hospital, CorsoSvizzera 164, 10149, Turin, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: De Avolio
mesh_terms: D019380:Anti-HIV Agents / Q000097:blood / Q000737:chemistry; D044966:Anti-Retroviral Agents / Q000097:blood* / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000069547:Cobicistat / Q000097:blood / Q000737:chemistry; D006575:Heterocyclic Compounds, 3-Ring / Q000097:blood / Q000737:chemistry; D006801:Humans; D057230:Limit of Detection; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D015363:Quinolones / Q000097:blood / Q000737:chemistry; D015203:Reproducibility of Results; D000068696:Rilpivirine / Q000097:blood / Q000737:chemistry; D019438:Ritonavir / Q000097:blood / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000099226:Dolutegravir
chemical_list: D019380:Anti-HIV Agents; D044966:Anti-Retroviral Agents; D000069547:Cobicistat; D006575:Heterocyclic Compounds, 3-Ring; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D015363:Quinolones; D000068696:Rilpivirine; D019438:Ritonavir; D000099226:Dolutegravir; C509700:elvitegravir
keywords: Antiretrovirals; HAART; Liquid chromatography; Protein precipitation; Reverse phase; Tandem mass spectrometry