Octenyl succinic anhydride (OSA)-modified starches have gained widespread interest, but the modification can produce two starches with different states ignored. Herein, the two types of starches, esterified starch (ES) and pregelatinized esterified starch (PES), prepared by OSA modification were separated, and their structural and functional characteristics were comprehensively explored. Results showed that compared with native starch (NS), ES and PES exhibited high water-holding capacity, solubility, and swelling power and significantly decreased pasting temperature and thermal stability. Dynamic rheological tests illustrated that OSA modification changed the rheological behavior of starches. Fourier transform infrared spectroscopy confirmed that PES with higher degree of substitution showed more obvious ester carbonyl and carboxylate groups than ES. Laser confocal micro-Raman spectroscopy revealed that the short-range molecular order of ES, especially PES, decreased after modification. X-ray diffraction indicated that OSA modification disrupted the crystalline structure of starch, and that more amylose-lipid complex was formed in PES. Scanning electron microscopy showed that OSA modification eroded starchs surface and reduced its smoothness, and significantly disrupted PES integrity. ES and PES could be developed as food additives for retrogradation inhibition of dough. These results provide new insights into OSA modification and expand its functional application in foods.