A validated assay by liquid chromatography-tandem mass spectrometry for the simultaneous quantification of elvitegravir and rilpivirine in HIV positive patients.

摘要: Because of the large variability in pharmacokinetics anti-HIV drugs, therapeutic drug monitoring patients may contribute to optimize overall efficacy and safety antiretroviral therapy. An LC–MS/MS method for simultaneous assay plasma novel agents rilpivirine (RPV) elvitegravir (EVG) has been developed that endeavor. Plasma samples (100 μL) extraction is performed by protein precipitation with acetonitrile, supernatant subsequently diluted 1:1 20-mM ammonium acetate/MeOH 50:50. After reverse-phase chromatography, quantification RPV EVG, using matrix-matched calibration samples, electrospray ionization–triple quadrupole mass spectrometry selected reaction detection positive mode. The stable isotopic-labeled compounds RPV-13C6 EVG-D6 were used as internal standards. The was validated according FDA recommendations, including assessment yield, matrix effects (<6.4%), well EVG short long-term stability plasma. Calibration curves over clinically relevant concentrations ranging from 5 2500 ng/ml 50 5000 ng/ml EVG. precise (inter-day CV%: 3–6.3%) accurate (3.8–7.2%). found be (<15%) all considered conditions (RT/48 h, +4°C/48 h, −20°C/3 months 60°C/1 h). Selected metabolite profiles analysis patients' revealed presence glucuronide, separated parent allowing exclude potential interferences through in-source dissociation glucuronide drug. This new, rapid robust LCMS/MS these two major new drugs offers an efficient analytical tool clinical studies routine service.