Sample collection, analytical methods, and pharmacokinetic analysis
Blood samples were collected for pharmacokinetics (PK) analysis of oral midazolam, prednisolone, and atorvastatin at 0, 0.5, 1, 2, 3, 4, 6, 8, 10, 12, 24 h post dose on days -1, 7, 14, 28, and 42. For PK analysis of intravenous midazolam, blood samples were collected at 0, 0.08, 0.17, 0.33, 0.5, 0.75, 1,1.5, 2, 4, 6, 8, 10, 12, and 24 h post dose on days -1, 7, 14, 28, and 42. Plasma concentrations of all analytes were measured by validated methods using liquid chromatography-tandem mass spectrometry. The linear analytical range of the assay was 20 − 20000 pg mL-1 for midazolam, 5 ­− 250 ng mL-1for prednisolone, and 50 − 10000 pg mL-1 for atorvastatin.
The primary PK parameters analyzed for midazolam, prednisolone, and atorvastatin included the AUC0-t, AUC from time of dosing to infinity (AUC0‐inf), and Cmax. Additional PK parameters included Tmax, and elimination half‐life (t1/2).