Organic Anion-Transporting Polypeptides (OATPs) are known to control the liver uptake of many drugs. Non-hepatic expression of OATPs has been reported although functional importance for whole-body pharmacokinetics (WBPK) remains unknown. Glyburide is a well described substrate of several hepatic and non-hepatic OATPs. Dynamic whole-body positron emission tomography (DWB-PET) with 11Cglyburide was performed in humans for determination of the importance of OATPs for liver uptake and WBPK. Seven healthy male subjects (24.7 ± 3.2 years) underwent 11Cglyburide PET scan with concomitant blood sampling. All subjects underwent baseline 11Cglyburide PET scan. Five subjects underwent a subsequent 11Cglyburide PET scan after infusion of the potent OATP inhibitor rifampicin (9 mg/kg i.v.). The transfer constant (kuptake) of 11Cglyburide from blood to the liver was estimated using the integration plot method. The tissue exposure of 11Cglyburide was described by the area under the time-activity curve (AUC) and corresponding tissue/blood ratio (AUCR). 11Cglyburide was barely metabolized in both the baseline and rifampicin conditions. Parent (unmetabolized) 11Cglyburide accounted for > 90 % of the plasma radioactivity. Excellent correlation was found between radioactive counting in arterial blood samples and in the image-derived input function, in both the baseline and rifampicin conditions (R2 = 97.9 %, p < 0.01). 11Cglyburide predominantly accumulated in the liver. Rifampicin decreased liver kuptake by 77.3 ± 7.3 %, which increased exposure in blood, kidneys, spleen, myocardium and brain (p < 0.05). No significant change in AUCR was observed except in the liver (p < 0.01). 11Cglyburide benefits from metabolic stability and high sensitivity to OATP inhibition which enables quantitative determination of OATP function. DWB-PET suggests negligible role for non-hepatic OATPs in controlling the tissue distribution of 11Cglyburide. Display omitted •Organic Anion-Transporting Polypeptides (OATP) form a family of influx transporters.•Importance of hepatic and non-hepatic OATP on drug delivery in humans is not known.•Whole-body PET imaging using the OATP substrate 11C-glyburide was performed in humans.•11C-glyburide benefits from unmet imaging properties to explore OATP function in humans.•PET data suggest a liver-selective importance of OATP in controlling drug delivery.