Simultaneous quantification of intracellular lamivudine and abacavir triphosphate metabolites by LC –MS/MS

Publication date: 10 May 2018 Source:Journal of Pharmaceutical and Biomedical Analysis, Volume 153 Author(s): Nagsen Gautam, Zhiyi Lin, Mary G Banoub, Nathan A Smith, Audai Maayah, JoEllyn McMillan, Howard E. Gendelman, Yazen Alnouti Nucleoside reverse transcriptase inhibitors (NRTIs) require intracellular phosphorylation to active triphosphate (TP) nucleotide metabolites before they can inhibit the HIV reverse transcriptase. However, monitoring these pharmacologically active TP metabolites is challenging due to their instability and their low concentrations at the pg/ml levels in blood and tissues. The combination of lamivudine (3TC) and abacavir (ABC) is one of the first lines for HIV therapy. Therefore, a sensitive, selective, accurate, and precise LC–MS/MS method was developed and validated for the simultaneous quantification of 3TC- and ABC-TP metabolites in mouse blood and tissues. Calibration curves were linear over the range of 10–100,000 pg/ml for 3TC-TP and 4–40,000 pg/ml for carbovir-TP (CBV-TP; phosphorylated metabolite of ABC). This corresponds to 2.1–21,322 fmol/106 cells for 3TC-TP and 0.8–8000 fmol/106 cells for CBV-TP. Accuracy and precision were less than 15% for all quality control sample (QCs), and absolute extraction recovery of were >65% for 3TC-TP and >90% for CBV-TP. The method was optimized to ensure stability of TP samples and standards during sample collection, preparation, analysis, and storage conditions. Th...
Source: Journal of Pharmaceutical and Biomedical Analysis - Category: Drugs & Pharmacology Source Type: research