In this work, cobalt phosphide nanoparticles (Co₂P NPs) were prepared by simple and mild hydrothermal method without the use of harmful phosphorous source. The morphological structure and surface component of Co₂P were characterized by transmission electron microscopy, X-ray diffraction and X-ray photoelectron spectroscopy measurements. Considering the excellent electrocatalytic reduction activity and good electrical conductivity of transition-metal phosphide, we fabricated Co₂P NPs on indium tin oxide (ITO) substrate (Co₂P/ITO) for H₂O₂ detection. The Co₂P/ITO transducer displayed a rapid amperometric response less than 5 s, a broader response range from 0.001 to 10.0 mM and a low detection limit of 0.65 μM. In addition, the non-enzymatic Co₂P/ITO sensor showed outstanding selectivity, reproducibility, repeatability and stability, all of which qualified the Co₂P/ITO electrode for quite a reliable and promising biosensor for H₂O₂ sensing.