title: High performance liquid chromatography using UV detection for the simultaneous quantification of the new non-nucleoside reverse transcriptase inhibitor etravirine (TMC-125), and 4 protease inhibitors in human plasma.
abstract: Etravirine (TMC-125, ETV) is a second-generation non-nucleoside reverse transcriptase inhibitor (NNRTI) that demonstrates potent activity against NNRTI-resistant strains of human immunodeficiency virus type-1 (HIV-1). Thus, ETV has been used in combination with ritonavir-boosted protease inhibitor (PI) and integrase inhibitor for therapy-experienced HIV-1-infected patients. On the other hand, as ETV is a substrate and inducer of cytochrome P450 3A4 (CYP3A4), ETV may induce metabolism of PI and alter the concentrations of co-administered PIs. In order to ensure optimal drug efficacy and prevention of resistance, it is essential to monitor plasma concentrations of ETV and PIs. Here we describe the application of HPLC with UV detection for the simulataneous assay of ETV and 4 PIs, darunavir (DRV), atazanavir (ATV), ritonavir (RTV) and lopinavir (LPV). In this study, the calibration curve of each drug was linear with the average accuracy ranging from 93.6 to 110.9%. Both intra- and interday coefficients of variation for each drug were less than 11.6%. The mean recovery of all drugs ranged from 88.0 to 97.5%. The limit of quantification was 0.04, 0.04, 0.04, 0.05 and 0.07 microg/ml for ETV, DRV, ATV, RTV and LPV, respectively. These results demonstrate that our HPLC-UV method can be used for routine determination of plasma concentrations of ETV and 4 PIs in clinical settings.
authors.affiliation: Department of Pharmacy, National Hospital Organization Nagoya Medical Center, Nagoya, Aichi 460-0001, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Hirano
authors.forename: Masaaki
authors.initials: M
authors.lastname: Takahashi
authors.forename: Eri
authors.initials: E
authors.lastname: Kinoshita
authors.forename: Masaaki
authors.initials: M
authors.lastname: Shibata
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Nomura
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Yokomaku
authors.forename: Motohiro
authors.initials: M
authors.lastname: Hamaguchi
authors.forename: Wataru
authors.initials: W
authors.lastname: Sugiura
mesh_terms: D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015658:HIV Infections / Q000097:blood* / Q000188:drug therapy; D017320:HIV Protease Inhibitors / Q000097:blood* / Q000627:therapeutic use; D015497:HIV-1 / Q000187:drug effects; D006801:Humans; D009570:Nitriles; D011724:Pyridazines / Q000097:blood* / Q000627:therapeutic use; D011743:Pyrimidines; D012015:Reference Standards; D015203:Reproducibility of Results; D018894:Reverse Transcriptase Inhibitors / Q000097:blood* / Q000627:therapeutic use; D013056:Spectrophotometry, Ultraviolet
chemical_list: D017320:HIV Protease Inhibitors; D009570:Nitriles; D011724:Pyridazines; D011743:Pyrimidines; D018894:Reverse Transcriptase Inhibitors; C451734:etravirine