The methods of measuring effective renal plasma flow (ERPF) based on the two-compartment analysis of plasma clearance of I-123-orthoiodohippurate (OIH) disregard the distribution of I-123-OIH to erythrocytes, resulting in over-estimation of ERPF. In this study, a new simple modification was established to obtain accurate ERPF. Intercompartmental clearance rates among plasma, erythrocytes, and extracellular fluid were determined through in vitro and in vivo experiments to enable the three-compartment analysis of plasma clearance curves of I-123-OIH using multiple blood samplings in comparison with the two-compartment analysis. Multiplying renal blood flow derived from two-compartment analysis of the whole blood clearance curve by (1-hematocrit/100) was a new modification for measuring ERPF to reduce the complexity of three-compartment analysis and to supplement the distribution of I-123-OIH to erythrocytes. Comparing ERPF derived from the modified two-compartment analysis with that from three-compartment analysis demonstrated good agreement (r = 0.883, p = 0.001) between the two methods. ERPF derived by this new method was also compared with the clearance of paraaminohippurate (PAH), resulting in a regression line which was much closer to unity and a better correlation between them (r = 0.926) than that between PAH clearance and ERPF (r = 0.909) derived from conventional two-compartment analysis.