Isosteviol has been reported to reverse hypertrophy and related inflammatory responses in in vitro models representative of cardiac muscle cells. The disposition of isosteviol is, however, characterized by secondary peaks and long plasma residence time despite reports of a relatively short half-life in liver fractions. The present study describes a compartmental approach to modelling the secondary peaks characteristic of isosteviol's concentration-time data in Sprague-Dawley rats. Oral (4 mg/kg) and intravenous (4 mg/kg) doses of isosteviol were administered to male and female Sprague-Dawley rats. Plasma samples collected between 0 and 72 h, and total bile secreted in 24 h, were analysed for isosteviol content with LC-MS/MS techniques. The disposition of isosteviol was, thereafter, described with a structural model that accounted for the sampling, liver and biliary secretion compartments, with a gap-time characterizing the accumulation and subsequent emptying of isosteviol for re-absorption. The half-life of isosteviol following oral dosing was about 103% greater in female rats than in the male, and the model-derived area under the concentration-time curve (AUC) in 72 h was about 756% greater in female animals than in males. Following the administration of intravenous doses of isosteviol, half-life and AUC in 24 h were about 332% and 595%, respectively, higher in female rats than in males. Isosteviol equivalent secreted into bile over 24 h accounted for about 94% of orally administered dose in male rats, and about 59% of oral dose in females. These findings show a differential systemic removal of isosteviol in Sprague-Dawley rats, likely explainable by gender-related differences in the glucuronidation-capacity of isosteviol.