4.6. Mouse Pharmacokinetic Study

Pharmacokinetic study in male mice (C57BL6J, Harlan Laboratories, Indianapolis, IN, USA) was run with 36 animals in each group at a GE dose of 250 mg/kg with a dose volume of 10 mL/kg. One group was given GE alone and in the second group GE was co-administered with KTZ (50 mg/kg). A terminal sampling design was used to collect blood samples at 0 (pre-dose), 0.08, 0.16, 0.25, 0.5, 1, 2, 3, 4, 6, 8, and 12 h. At each time, 1 mL of blood was collected in ethylenediaminetetraacetic acid di-potassium salt (K₂EDTA) tubes (BD Biosciences, Franklin Lakes, NJ, USA). Plasma was separated by centrifugation of samples at 4000 rpm for 10 min and stored below −60 °C till bioanalysis. All experiments were performed with approval from the Institutional Animal Care and Use Committee (IACUC, approval code: A14031). All animals were acclimatized for five days before dosing in the experimental area. Food and water was provided ad libitum throughout the study period. Animals were marked and housed (three per cage, n = 36 per group) in polypropylene cages and maintained in controlled environmental conditions with 12 h light and dark cycles. The temperature and humidity of the room was maintained between 22 ± 3 °C and 30% to 50%, respectively, and approximately 10–12 fresh air change cycles per hour.