Plasma samples were processed following a similar protocol.32 (link) These samples were injected on the Hypersil GOLD C18 column (1.8 μm, 2.1mm × 50 mm) with UHPLC ultimate 3000 from Dionex coupled with a TSQ Vantage mass spectrometer coupled with a UHPLC ultimate 3000 from Dionex to quantitate the concentration of the main metabolites of PI-103BE (9 ) and PI-103 (7 ). The 10 μL samples were run with the gradient starting at 0.6 mL/min from 10% mobile phase B (Acetonitrile with 0.05% formic acid) and 90% mobile phase A (water with 0.05% formic acid) until 0.5 min, up to 100% B at 3 min, and until 6 min, then came back to 10% B until equilibration. The TSQ Vantage was set at spray vantage at 3200 V, vaporizer temperature at 365 °C, Sheath gas pressure at 42 PSI, Aug Gas pressure at 12 psi, Capillary temperature at 350 °C.
All procedures involving the animals were conducted in compliance with State and Federal laws, standards of the U.S. Department of Health and Human Services, and guidelines established by Xavier University Animal Care and Use Committee. The facilities and laboratory animals program of Xavier University are accredited by the Association for the Assessment and Accreditation of Laboratory Animal Care.
All procedures involving the animals were conducted in compliance with State and Federal laws, standards of the U.S. Department of Health and Human Services, and guidelines established by Xavier University Animal Care and Use Committee. The facilities and laboratory animals program of Xavier University are accredited by the Association for the Assessment and Accreditation of Laboratory Animal Care.