Iron toxicity is the major cause of tissue damage in patients with iron overload. Iron deposits mainly in the liver, where its concentration closely correlates with whole body iron overload. Different techniques have been proposed for estimating iron content, with liver biopsy being the gold standard despite its invasiveness and influence by sampling error. Recently, magnetic resonance imaging (MRI) has been established as an effective technique for evaluating iron overload by measuring T2∗ in the liver. However, various factors associated with the adopted analysis technique, mainly the exponential fitting model and signal averaging method, affect the resulting measurements. In this study, we evaluate the influences of these factors on T2∗ measurement in numerical phantom, calibrated phantoms, and nine patients with different degrees of iron overload. The results show different performances among the fitting models and signal averaging methods, which are affected by SNR, image quality and signal homogeneity inside the selected ROI for analysis.