Validation of a UHPLC-MS/MS Method to Quantify Twelve Antiretroviral Drugs within Peripheral Blood Mononuclear Cells from People Living with HIV


De Nicolo, Amedeo, Ianniello, Alice, Ferrara, Micol, Avataneo, Valeria, Cusato, Jessica, Antonucci, Miriam, De Vivo, Elisa, Waitt, Catriona ORCID: 0000-0003-0134-5855, Calcagno, Andrea, Trentalange, Alice et al (show 4 more authors) (2021) Validation of a UHPLC-MS/MS Method to Quantify Twelve Antiretroviral Drugs within Peripheral Blood Mononuclear Cells from People Living with HIV PHARMACEUTICALS, 14 (1). pp. 1-16. ISSN 1424-8247, 1424-8247

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Abstract

Recently, anti-HIV treatment has achieved high efficacy and tolerability. Nevertheless, few data are available about the intracellular penetration of antiretrovirals, partly due to the tech-nical challenges related to intracellular quantification. This work aimed to validate an ultra-high performance liquid chromatography (UHPLC) tandem mass spectrometry (MS/MS) method for the simultaneous quantification of maraviroc, nevirapine, rilpivirine, dolutegravir, raltegravir, cobicis-tat, darunavir, ritonavir, atazanavir, efavirenz, elvitegravir, and etravirine within peripheral blood mononuclear cells (PBMCs) and apply it to samples from patients. PBMCs were isolated by density gradient on cell preparation tubes (CPT). Samples were prepared by addition of internal standards (IS), sonication, centrifugation, and drying. Reconstituted extracts underwent chromatographic separation by reversed phase UHPLC and detection was performed by electrospray ionization and multiple reaction monitoring. Method validation followed FDA and EMA guidelines, showing ac-ceptable accuracy, precision, recovery and IS-normalized matrix effect. The application to 56 samples from patients undergoing antiretroviral treatment provided description of intracellular penetration, showing method eligibility for future studies.

Item Type: Article
Uncontrolled Keywords: liquid chromatography, tandem mass spectrometry, peripheral blood mononuclear cells, dolutegravir, intracellular
Depositing User: Symplectic Admin
Date Deposited: 11 Jan 2021 11:19
Last Modified: 16 Jun 2026 12:22
DOI: 10.3390/ph14010012
Open Access URL: https://www.mdpi.com/1424-8247/14/1/12/pdf
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URI: https://livrepository.liverpool.ac.uk/id/eprint/3112496
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