title: Simultaneous determination of the antiretroviral agents: amprenavir, lopinavir, ritonavir, saquinavir and efavirenz in human peripheral blood mononuclear cells by high-performance liquid chromatography-mass spectrometry.
abstract: A selective and accurate assay for the simultaneous quantitation of four protease inhibitors (PIs) (amprenavir (APV), lopinavir (LPV), ritonavir (RTV) and saquinavir (SQV)) and a non-nucleoside reverse transcriptase inhibitor (NNRTI) (efavirenz, EFV) in human peripheral blood mononuclear cells using high-performance liquid chromatography-mass chromatography (LC/MS) has been developed and validated. After liquid-liquid extraction, the antiretroviral agents were separated within 15 min. The calibration curves of each drug showed a good linearity in a range of concentration between 2 and 200 ng/3 x 10(6) cells for amprenavir, lopinavir, efavirenz, 1.60 and 128 ng/3 x 10(6) cells for ritonavir and saquinavir. Mean intra- and inter-assay coefficients of variation over the ranges of the standard curves were less than 15% and mean extraction recoveries ranged 88.7-112.1%. The limits of quantification were 2 ng/3 x 10(6) cells for amprenavir, lopinavir, efavirenz, 1 ng/3 x 10(6) cells for ritonavir and 1.6 ng/3 x 10(6) cells for saquinavir. This novel LC/MS assay, which provides an excellent method for simultaneous intra-cellular determination of amprenavir, lopinavir, ritonavir, saquinavir and efavirenz in human peripheral blood mononuclear cells, could be successfully applied for therapeutic drug monitoring and pharmacokinetic studies.
authors.affiliation: Clinical Pharmacokinetic Department EA 525, Victor Segalen Bordeaux 2 University, Bordeaux, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Rouzes
authors.forename: Karine
authors.initials: K
authors.lastname: Berthoin
authors.forename: Fabien
authors.initials: F
authors.lastname: Xuereb
authors.forename: Sarah
authors.initials: S
authors.lastname: Djabarouti
authors.forename: Isabelle
authors.initials: I
authors.lastname: Pellegrin
authors.forename: Jean Luc
authors.initials: JL
authors.lastname: Pellegrin
authors.forename: Anne Cécile
authors.initials: AC
authors.lastname: Coupet
authors.forename: Sylvie
authors.initials: S
authors.lastname: Augagneur
authors.forename: Hélène
authors.initials: H
authors.lastname: Budzinski
authors.forename: Marie Claude
authors.initials: MC
authors.lastname: Saux
authors.forename: Dominique
authors.initials: D
authors.lastname: Breilh
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017320:HIV Protease Inhibitors / Q000097:blood*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D017320:HIV Protease Inhibitors