Development and Validation of an Up-to-Date Highly Sensitive UHPLC-MS/MS Method for the Simultaneous Quantification of Current Anti-HIV Nucleoside Analogues in Human Plasma.
Amedeo De NicolòAlessandra MancaAlice IannielloAlice PalermitiAndrea CalcagnoMicol FerraraMiriam AntonucciJessica CusatoValeria AvataneoElisa Delia De VivoStefano BonoraFrancesco Giuseppe De RosaGiovanni Di PerriAntonio D'AvolioPublished in: Pharmaceuticals (Basel, Switzerland) (2021)
Therapeutic options to treat HIV infection have widened in the past years, improving both effectiveness and tolerability, but nucleoside reverse transcriptase inhibitors (NRTIs) are still considered the standard backbone of the combination regimens. Therapeutic drug monitoring (TDM) can be useful for these drugs, due to concentration-effect relationship, with risk of ineffectiveness, toxicity or adherence concerns: in this scenario, robust and multiplexed methods are needed for an effective TDM activity. In this work, the first validated ultra-high spectrometry liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) method is described for the high-sensitive simultaneous quantification of all the currently used NRTIs in human plasma, including tenofovir alafenamide (TAF), following FDA and EMA guidelines. The automated sample preparation consisted in the addition of an internal standard (IS) working solution, containing stable-isotope-linked drugs, protein precipitation and drying. Dry extracts were reconstituted with water, then, these underwent reversed phase chromatographic separation: compounds were detected through electrospray ionization and multiple reaction monitoring. Accuracy, precision, recovery and IS-normalized matrix effect fulfilled guidelines' requirements. The application of this method on samples from people living with HIV (PLWH) showed satisfactory performance, being capable of quantifying the very low concentrations of tenofovir (TFV) in patients treated with TAF.
Keyphrases
- tandem mass spectrometry
- liquid chromatography
- ultra high performance liquid chromatography
- simultaneous determination
- ms ms
- high resolution mass spectrometry
- antiretroviral therapy
- solid phase extraction
- gas chromatography
- high performance liquid chromatography
- mass spectrometry
- liquid chromatography tandem mass spectrometry
- high resolution
- molecularly imprinted
- hiv infected
- hiv positive
- human immunodeficiency virus
- clinical practice
- deep learning
- randomized controlled trial
- hiv aids
- high throughput
- systematic review
- hiv testing
- small molecule
- hepatitis c virus
- single cell
- metabolic syndrome
- open label
- glycemic control
- insulin resistance
- protein protein
- amino acid
- oxidative stress