Three maize cultivars with different endosperm types (flint, semiflint, and dent maize cultivars) were studied to characterize vitreous endosperm flour properties compared with those of floury endosperm flour from the same maize kernels. Vitreous endosperm flour had higher amylose and protein contents, and lower starch content, higher percentage of large starch granule, bigger mean diameter of starch granule, higher iodine capacity, higher trough viscosity and final viscosity and setback, lower swelling power, lower peak viscosity and breakdown, and higher peak time and pasting temperature than the counterpart floury endosperm flour. X‐ray diffraction analysis indicated typical A‐pattern for starches of vitreous and floury endosperm flours. Floury endosperm flour showed the presence of greater crystallinity and higher enthalpy change (
∆Hgel
) than vitreous endosperm flour for three cultivars. Retrogradation enthalpy (
∆Hret
) and retrogradation percentage (
R
) of vitreous and floury endosperm flours ranged from 6.23 to 7.92 J/g and 52.72% to 73.62%, and from 5.46 to 6.45 J/g and 45.70% to 56.58%, respectively. In conclusion, vitreous and floury endosperm flours had significantly different physicochemical properties. Results of this study provide a foundation for better and valid utilization of different endosperm section during grain processing.