The transport kinetics of [99mTc]-pyridoxyl-5-methyltryptophan were studied by three-compartment model analysis for hepatobiliary scintigraphy in 45 patients with chronic viral liver diseases. Three-compartment model analysis was studied using the time-activity curves of the regions of the heart, liver, and biliary tract and intestine (excretory compartment). The k12 (hepatic uptake rate constant), k21 (hepatic efflux rate constant), and ke1 (hepatic excretion rate constant) were calculated by the nonlinear least-squares method. Among the three parameters obtained by model analysis, k12 values more prominently differed among diseases and correlated well with blood tests such as total bilirubin, total bile acids, or 15 min retention of ICG. In conclusion, three-compartment model analysis of the hepatic handling of [99mTc]-pyridoxyl-5-methyltryptophan is useful in evaluating hepatic transport function. k12 is the most sensitive parameter for this.