The physicochemical properties of high-pressure homogenized (microfluidized) maize starch with different preheating temperatures (50, 60oC), levels of pressure (34.5, 69, 138 MPa), and numbers of pass (1, 2, 3 pass) were examined in this study. The enzyme susceptible starch (ESS) content, morphological property, X-ray diffraction, and Rapid Visco Analyzer (RVA) profile of starch were significantly altered via increasing the number of passes and preheating temperatures. The amount of ESS and the diffraction pattern of starch indicated that the granular crystalline structure of starch was severely damaged by increasing the number of passes and preheating temperatures. The morphology of starch granule was changed from angular to spherical shape with the damaged surface as the pressure increased. Moreover, damaged starch particles gathered to form a larger mass when treated at a higher temperature with the increasing number of passes, indicating that different types and extents of damage occurred. The RVA profile of starch showed a moderate peak viscosity with increased pasting stability against shear thinning similar to that of cross-linked starch as the number of passes and preheating temperatures increased. These results suggested that microfluidization combined with preheating might be used as a potential alternative method for the modification of starch such as cross-linked starch.