Conventional 2-dimensional planar imaging of 123 I-metaiodobenzylguanidine ( 123 I-mIBG) is not fully quantitative. To develop a more accurate quantitative imaging approach, we investigated dynamic SPECT imaging with kinetic modeling in healthy humans to obtain the myocardial volume of distribution (V T ) for 123 I-mIBG. Methods: Twelve healthy humans underwent 5 serial 15-min SPECT scans at 0, 15, 90, 120, and 180 min after bolus injection of 123 I-mIBG on a hybrid cadmium zinc telluride SPECT/CT system. Serial venous blood samples were obtained for radioactivity measurement and radiometabolite analysis. List-mode data of all the scans were binned into frames and reconstructed with attenuation and scatter corrections. Myocardial and blood-pool volumes of interest were drawn on the reconstructed images to derive the myocardial timeactivity curve and input function. A population-based blood-to-plasma ratio (BPR) curve was generated. Both the population-based metabolite correction (PBMC) and the individual metabolite correction (IMC) curves were generated for comparison. V T values were obtained from different compartment models, using different input functions with and without metabolite and BPR corrections. Results: The BPR curve reached the peak value of 2.1 at 13 min after injection. Parent fraction was approximately 58% ± 13% at 15 min and stabilized at approximately 40% ± 5% by 180 min after injection. Two radiometabolite species were observed. When the reversible 2-tissue-compartment fit was used, the mean V T value was 29.0 ± 12.4 mL/cm 3 with BPR correction and PBMC, a 188% ± 32% increase compared with that without corrections. There was significant difference in V T with BPR correction (P 5 2.3e-04) as well as with PBMC (P 5 1.6e-05). The mean difference in V T between PBMC and IMC was −3% ± 8%, which was insignificant (P 5 0.39). The intersubject coefficients of variation after PBMC (43%) and IMC (42%) were similar. Conclusion: The myocardial V T of 123 I-mIBG was established in healthy humans for the first time. Accurate kinetic modeling of 123 I-mIBG requires both BPR and metabolite corrections. Population-based BPR correction and metabolite correction curves were developed, allowing more convenient absolute quantification of dynamic 123 I-mIBG SPECT images. Met aiodobenzylguanidine labeled with 123 I ( 123 I-mIBG) is a norepinephrine analog that has a reuptake mechanism similar to that of norepinephrine without being catabolized in the myocardial sympathetic nerve endings and has been the most widely used sympathetic innervation imaging agent in research and clinical studies for risk stratification of patients with congestive heart failure (HF) (1-5). For conventional 123 I-mIBG imaging, early (15-30 min) and delayed (3-5 h) 2-dimensional (2D) planar images are typically acquired and used for calculating heart-to-mediastinum ratio (HMR) and washout rate (WOR). However, 2D planar imaging is not fully quantitative, because of the superposition of background structures with the 2D heart region of intere...