title: Validation of a fast method for quantitative analysis of elvitegravir, raltegravir, maraviroc, etravirine, tenofovir, boceprevir and 10 other antiretroviral agents in human plasma samples with a new UPLC-MS/MS technology.
abstract: Therapeutic drug monitoring (TDM) of antiretrovirals requires accurate and precise analysis of plasma drug concentrations. This work describes a simple, fast and sensitive UPLC-MS/MS method for determination of the commonly used protease inhibitors such as amprenavir, atazanavir, darunavir, indinavir, lopinavir, ritonavir, saquinavir and tipranavir, tenofovir a nucleoside reverse transcriptase inhibitor (NRTI), the non-NRTI such as efavirenz, nevirapine, etravirine, the CCR5 antagonist maraviroc as well as the more recent antiretrovirals, the integrase inhibitors such as raltegravir, elvitegravir and the new direct acting anti-HCV boceprevir. Adapted deuterated internal standard was added to plasma aliquots (100μl) prior to protein precipitation with methanol and acetonitrile. This method employed ultra-performance liquid chromatography coupled to tandem mass spectrometry with electrospray ionization mode. All compounds eluted within 4.2-min run time. Calibration curves were validated, with correlation coefficients (r(2)) higher than 0.997, for analysis of therapeutic concentrations reported in the literature. Inter- and intra-assay variations were <15%. Evaluation of accuracy shows a deviation <15% from target concentration at each quality control level. No significant matrix effect was observed for any of the antiretroviral studied. This new validated method fulfills all criteria for TDM of 15 antiretrovirals and boceprevir drugs and was successfully applied in routine TDM of antiretrovirals.
authors.affiliation: Department of Pharmacology, E.A.3801, SFR CAP-santé, Reims University Hospital, 51, rue Cognacq-Jay, 51095 Reims Cedex, France. Electronic address: zoubir.djerada@univ-reims.fr.
authors.forename: Zoubir
authors.initials: Z
authors.lastname: Djerada
authors.forename: Catherine
authors.initials: C
authors.lastname: Feliu
authors.forename: Claire
authors.initials: C
authors.lastname: Tournois
authors.forename: Damien
authors.initials: D
authors.lastname: Vautier
authors.forename: Laurent
authors.initials: L
authors.lastname: Binet
authors.forename: Arnaud
authors.initials: A
authors.lastname: Robinet
authors.forename: Hélène
authors.initials: H
authors.lastname: Marty
authors.forename: Claire
authors.initials: C
authors.lastname: Gozalo
authors.forename: Denis
authors.initials: D
authors.lastname: Lamiable
authors.forename: Hervé
authors.initials: H
authors.lastname: Millart
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000097:blood; D044966:Anti-Retroviral Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D003510:Cyclohexanes / Q000097:blood; D006801:Humans; D000077592:Maraviroc; D013058:Mass Spectrometry / Q000379:methods / Q000592:standards; D009570:Nitriles; D063065:Organophosphonates / Q000097:blood; D011392:Proline / Q000031:analogs & derivatives / Q000097:blood; D011724:Pyridazines / Q000097:blood; D011743:Pyrimidines; D011760:Pyrrolidinones / Q000097:blood; D015363:Quinolones / Q000097:blood; D000068898:Raltegravir Potassium; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards*; D000068698:Tenofovir; D013997:Time Factors; D014230:Triazoles / Q000097:blood
chemical_list: D044966:Anti-Retroviral Agents; D003510:Cyclohexanes; D009570:Nitriles; D063065:Organophosphonates; D011724:Pyridazines; D011743:Pyrimidines; D011760:Pyrrolidinones; D015363:Quinolones; D014230:Triazoles; C451734:etravirine; D000068898:Raltegravir Potassium; C509700:elvitegravir; C512204:N-(3-amino-1-(cyclobutylmethyl)-2,3-dioxopropyl)-3-(2-((((1,1-dimethylethyl)amino)carbonyl)amino)-3,3-dimethyl-1-oxobutyl)-6,6-dimethyl-3-azabicyclo(3.1.0)hexan-2-carboxamide; D000068698:Tenofovir; D011392:Proline; D000225:Adenine; D000077592:Maraviroc
keywords: ARV; Antiretrovirals; Antiviral; Boceprevir; Ct; Drug monitoring; Elvitegravir; IS; MRM; TDM; UPLC-MS/MS; antiretroviral drugs; internal standard; multiple reaction transition; plasma trough concentration; therapeutic drug monitoring; ultra-performance liquid chromatography-tandem mass spectrometry