Male Sprague Dawley (SD) rats (average weighing 220–250) g were purchased from the laboratory animal center of Wenzhou Medical University, Zhejiang Province, China. Animals were maintained in a controlled environment with a 12 h light-dark cycle at 20°C–25°C and 55% ± 15% relative humidity. Diet intake was not allowed for a period of 12 h prior to the experiment. Then, water was allowed to be free and food was provided after the experiment. All the experimental procedures and protocols were reviewed and approved by the animal ethics committee of Wenzhou Medical University according to the guide for the care and use of laboratory animals (xmsq2021-0409). 36 male rats aged 8–10 weeks were randomly selected and divided into six groups for gavage. The group which was orally administered 40 mg/kg ketoconazole and 10 mg/kg tofacitinib acted as the positive control group and the group which was only orally administered 10 mg/kg tofacitinib acted as the negative group. Another four groups were orally administered 20 mg/kg bergapten, 50 mg/kg of bergapten, 20 mg/kg of isopsoralen and 50 mg/kg of isopsoralen, respectively, except 10 mg/kg of tofacitinib. Blood (300 μL) was collected from the tail veins into 1.5 mL tubes at 5, 15, and 30 min and 1, 2, 3, 4, 6, 8, 12, and 24 h after administering the drugs. The supernatant was collected and stored at −20°C after centrifugation at 4000 rpm for 10 min. The samples were restored to RT before analysis. 50 μL sample was accurately drawn to 1.5 mL of EP tube, 150 μL of midazolam internal standard solution (200 ng/mL) was added to the tube, then placed and vortexed for 15 s. After centrifugation at 13, 000 rpm for 15 min, 150 µL of supernatant was prepared for the UPLC-MS/MS system to analyze.
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