Rutin‐loaded octenylsuccinic anhydride (OSA)–starch nanospheres are successfully prepared by a dialysis embedding method with self‐assembled modified starch nanospheres as the carrier. The modification of the starch is performed using OSA. The formation and characteristics of the rutin‐loaded OSA–starch nanospheres are studied by scanning electron microscopy (SEM), fluorescence spectroscopy, and dynamic light scattering (DLS). The effects of the degree of substitution (DS) in the OSA–starch, the initial water content, and the rutin concentration on the morphology, particle size, encapsulation efficiency, and drug loading capacity of the nanospheres are evaluated. The SEM images show that the rutin‐loaded OSA–starch nanospheres, with good spherical shape, are larger than regular OSA–starch nanospheres. The drug loading, encapsulation efficiency, and particle size decrease as the degree of substitution or initial water content increases. With an increase of the rutin loading, the drug loading increases, while the encapsulation efficiency and particle size decrease. Rutin is successfully incorporated into the OSA–starch nanospheres. The OSA–starch nanospheres show good sustained release of rutin, providing a theoretical basis for the application of starch nanospheres in the field of medicine.