Contents lists available at ScienceDirect Journal of Chromatography B journal homepage: www.elsevier.com/locate/jchromb UHPLC-MS/MS assay for simultaneous determination of amlodipine, metoprolol, pravastatin, rosuvastatin, atorvastatin with its active metabolites in human plasma, for population-scale drug-drug interactions studies in people living with HIV Perrine Courlet a , Dany Spaggiari a,1 , Vincent Desfontaine a , Matthias Cavassini b , Susana Alves Saldanha a , Thierry Buclin a , Catia Marzolini c , Chantal Csajka a,d , Laurent-Arthur Decosterd a, ⁎ a Service of Clinical Pharmacology, Lausanne University Hospital and University of Lausanne, Switzerland b Service of Infectious Diseases, Lausanne University Hospital and University of Lausanne, Switzerland c Departments of Medicine and Clinical Research, University Hospital of Basel and University of Basel, Switzerland d School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Switzerland 1. Introduction Highly active antiretroviral treatments (ARTs) have transformed Human Immunodefciency Virus (HIV) infection from a deadly disease into a chronic condition. Consequently, people living with HIV (PLWH) live longer and the proportion of older individuals within the HIV-in- fected population is constantly growing [1]. Notably, interactions be- tween traditional health risk factors and HIV infection itself lead PLWH to a higher risk of adverse cardiovascular events [2,3]. A wide variety of cardiovascular drugs such as calcium channel blockers, β-blockers and statins, are consequently prescribed to PLWH along with their ARTs. Atorvastatin, rosuvastatin, and pravastatin are the most prescribed lipid-lowering agents in PLWH [4]. Atorvastatin is predominantly me- tabolized by cytochrome P450 3A4 (CYP3A4) into two major active metabolites (namely ortho-hydroxy-atorvastatin, and para-hydroxy- atorvastatin), and the inactive metabolite atorvastatin lactone [5]. Conversely, rosuvastatin and pravastatin undergo minor hepatic me- tabolism. However, all statins are actively transported in the liver by the organic anion transporting polypeptide 1B1 (OATP1B1) [6]. In the case of polymedicated PLWH, antiretroviral drugs (ARVs) such as ata- zanavir are strong inhibitors of OATP1B1 and are therefore expected to substantially increase statin exposure by both inhibiting the entry of the statin in the liver and by further inhibiting its biotransformation [7–9]. This drug-drug interaction (DDI) can lead to severe adverse drug re- actions such as rhabdomyolysis [10]. Additionally, 26.1% of PLWH have been reported to sufer from high blood pressure in the Swiss HIV Cohort Study (SHCS) [11]. Cal- cium channel blockers, such as amlodipine, are the preferred frst-line antihypertensive drugs for most patients, including PLWH [12]. Am- lodipine is predominantly metabolized by CYP3A4/5 [13,14]. In con- sequence, the inhibition of CYP3A4 by ritonavir-boosted protease in- hibitors (PIs) can increase amlodipine plasma concentrations, potentially leading to toxicity [13]. Finally, metoprolol, a β-blocker commonly used in the treatment of hypertension and heart failure, is predominantly metabolized by the polymorphic CYP2D6, and boosted ARVs with inhibitory properties for this isoform could potentially lead to increase metoprolol plasma concentrations, though reportedly to a moderate extent [15]. Cardiovascular drugs are commonly used in PLWH, particularly in the context of an aging population, and are subject to DDIs leading to substantial risks of iatrogenic toxicity. Bioanalytical methodologies are therefore needed to determine plasma concentrations of cardiovascular drugs in PLWH and help manage DDIs in clinical practice. Various methods have been published for the quantifcation in human plasma of a wide variety of cardiovascular drugs including calcium channel blockers, β-blockers and statins. Previously published assays for statins in human plasma, imply a chromatographic step on C18 support [16–29], or on Zorbax-SB phenyl column such as proposed by Macwan et al. for atorvastatin or rosuvastatin and their metabolites [30,31]. While multiplex liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analyses of various combinations of statins, calcium-channel inhibitors and β-blocker agents have been https://doi.org/10.1016/j.jchromb.2019.121733 Received 22 May 2019; Received in revised form 2 July 2019; Accepted 24 July 2019 ⁎ Corresponding author at: Laboratory of Clinical Pharmacology BU19-04, Service of Clinical Pharmacology, Centre Hospitalier Universitaire Vaudois, CHUV, 1011 Lausanne, Switzerland. E-mail addresses: perrine.courlet@chuv.ch (P. Courlet), dany.spaggiari@debiopharm.com (D. Spaggiari), vincent.desfontaine@chuv.ch (V. Desfontaine), matthias.cavassini@chuv.ch (M. Cavassini), susana.alves-saldanha@chuv.ch (S. Alves Saldanha), thierry.buclin@chuv.ch (T. Buclin), catia.marzolini@usb.ch (C. Marzolini), chantal.csajka@chuv.ch (C. Csajka), LaurentArthur.Decosterd@chuv.ch (L.-A. Decosterd). 1 Present address: Debiopharm International SA, Lausanne, Switzerland. Journal of Chromatography B 1125 (2019) 121733 Available online 25 July 2019 1570-0232/ © 2019 Elsevier B.V. All rights reserved. T