Simultaneous Quantitative Analysis of HIV Protease Inhibitors in Human Plasma Using Core-Shell Column and Fast MRM Detection

Abstract

To support therapeutic drug monitoring (TDM) a quantitative method for eight HIV protease inhibitors (PIs): nelfinavir, amprenavir, indinavir, saquinavir, atazanavir, darunavir, ritonavir and lopinavir in human plasma was developed. Separation of the analytes was achieved using core-shell column for UHPLC separation and mass spectrometric detection in fast multiple reaction monitoring (MRM) mode. Human plasma samples were collected on filter paper in the tube-based dried blood spot device and the analytes were extracted with methanol directly. Core-shell column and fast MRM allowed achieving an analysis time of six minutes. Using a plasma aliquot of 15 mu L a dynamic linear range of 25-5000 ng/mL (ritonavir), 25-10000 ng/mL (nelfinavir and amprenavir) and 25-20000 ng/mL (indinavir, saquinavir, atazanavir, darunavir and lopinavir) could be achieved with precision and accuracies better than 15 \%.