Ultrasmall superparamagnetic iron oxide (Fe3O4) nanoparticles coated by biocompatible pullulan (Pu‐USPIO) with sizes below 10 nm and having a magnetite core and a hydrophilic outer shell of pullulan were prepared. The formed Pu‐USPIOs were thoroughly characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, atomic force microscopy, and small‐angle X‐ray scattering experiments. The content of magnetic nanoparticles embedded into the pullulan matrix was determined by thermogravimetric analysis. Vibrating sample magnetometry analysis was used to evaluate the magnetic properties of the Pu‐USPIO samples. Because of the presence of pullulan, these nanoparticles could be conditioned in many versatile forms, from a clear solution to magnetic films, for potential applications, including magnetic hyperthermia mediators. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 42926.