As an adsorbent, natural pyrite was used to study adsorption characteristics of Rhodamine B (RhB), which was characterized by X-ray diffraction (XRD) and scanning electron microscope (SEM). In this study, the factors which affect the adsorption of RhB by pyrite such as reaction time, pH, temperature and ionic strength were investigated. The maximum RhB uptake was 21.3 mg/g, which was observed at the equilibrium time 120 min and pH 4.0. The adsorption capacity of RhB increased gradually with adsorption temperature, while the ionic strength showed little effect on the adsorption process. The adsorption process was well described by the Langmuir isotherm model and the pseudo second-order kinetics model. Thermodynamics results confirmed that the adsorption was a spontaneous process. RhB loaded pyrite was regenerated through calcination in air. The results show that the regenerated pyrite could effectively remove 50% of RhB, demonstrating a strong reuse potential.