HPLC-MS analysis for determination of plasma concentrations
The supernatant were analyzed using a liquid phase system, which included the Waters ACQUITY Ultra Performance Liquid Chromatography I-Class Plus system, equipped for the analysis of Bosutinib, Nintedanib, Rivaroxaban, Apixaban, and Vilazodone, as well as a High Performance Liquid Chromatography system (Shimadzu LC), used for the analysis of Fostamatinib, Ponatinib, Axitinib, Crizotinib, Venetoclax, Ibrutinib, Sonidegib, Roflumilast, Suvorexant, Apremilast, Elagolix, Istradefylline, Panobinostat, Rolapitant, Tasimelteon, Fedratinib, Lenvatinib, Pomalidomide, Ruxolitinib, Ospemifene, Edoxaban, Baricitinib, Vorapaxar, Flibanserin, Macitentan, Safinamide, and Tofacitinib. Separation of the analytes was achieved using a reversed-phase ACE T3 column (1.8 μm, 2.1×50 mm) and the mobile phase consisted of 0.1% formic acid in water for the A phase and 0.1% formic acid in acetonitrile for the B phase. The samples concentrations were then determined using a Triple Quad 6500+ mass spectrometer (Applied Biosystems, USA), equipped with electron spray ionization (ESI) and a tandem quadruple mass analyzer. Data processing was performed using Analyst software (SCIEX), and the mass spectrum parameters can be found in Table S1.