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The cassette dosing technique is employed in the drug discovery stage of non-clinical studies to obtain pharmacokinetic data from multiple drug candidates in a single experiment. The objective of the current investigation was to evaluate the effect of sex and food on the selected pharmacokinetic parameters of four biopharmaceutical classification system (BCS) drugs (BCS-I: propranolol, BCS-II: diclofenac, BCS-III: atenolol, and BCS-IV: acetazolamide) utilizing cassette dosing in male and female rats under fed and fasting conditions. Different animal groups were dosed intravenous (i.v) and oral at 1 and 10 mg/kg, respectively, in the form of cassette at a dose of 5 mL/kg. Blood samples were analyzed by liquid chromatography-tandem mass spectrometry. Pharmacokinetics parameters were calculated using Phoenix software version 8.1. A significant increase (p < 0.05) of the area under the plasma concentration–time (AUC0-last) was observed for diclofenac and acetazolamide in females over males after i.v dosing. Additionally, acetazolamide showed greater instantaneous concentration at the time of dosing, and clearance in females (p < 0.05) compared to males after i.v administration. After oral dosing, propranolol exhibited significant variations (p < 0.05) in the maximum drug concentration (Cmax), AUC0-last, the volume of distribution (Vd), and bioavailability in females as compared to males under fed state. Diclofenac showed significant changes (p < 0.05) in AUC0-last, and clearance (Cl) in females as compared to males under fasting and fed state. However, acetazolamide exhibited a significant enhancement (p < 0.05) in AUC0-last, Vd, and Cl in fasting females than the males. The data here illustrates that there is an appreciable difference in AUC and Cmax values exist in male and female rats under fed and fasting conditions administered with the cassette dosing of tested BCS class drugs.

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