An exhaustive characterization of the physicochemical properties of gum arabic (GA)-coated Fe3O4 magnetic nanoparticles was conducted in this work. These nanoparticles were obtained via the in-situ coprecipitation method (a fast single-step method) in two GA:Fe ratios, 10:1 and 20:1, respectively. Several experimental techniques were applied in the characterization process, all of them described below. Using Transmission Electron Microcopy images, they were shown to have spherical-like morphology with 11 nm diameter. The Fourier Transform Infrared spectra confirmed the attachment of the GA on the surface of the magnetic nanoparticles (MNPs), providing good colloidal stability from pH 7 to 8. The thickness of the coatings (1.7 nm and 1.1 nm) was determined using thermogravimetric measurements. A high specific absorption rate and superparamagnetic properties were determined using alternant and static magnetic fields, respectively. The GA-coated MNPs were non-cytotoxic, according to tests on HT-29 human intestine cells. Additionally, HT-29 cells were exposed to magnetic fluid hyperthermia at 530 kHz, and the induction of cell death by the magnetic field, due to the heating of GA-coated MNP, was observed.