For the first time silicon nitride (Si3N4) nanoparticles was used for preparation electrochemical biosensor. GOx immobilized on the Si3N4 nanoparticles exhibits facile and direct electrochemistry. The surface coverage and heterogeneous electron transfer rate constant (ks) of immobilized GOx were 6.31013 molcm2 and 47.40.3 s1. The sensitivity, linear concentration range and detection limit of the biosensor for glucose detection were 38.57 mAmM1cm2, 25 mM to 8 mM and 6.5 mM, respectively. This biosensor also exhibits good stability, reproducibility and long life time. These indicate Si3N4 nanoparticles is good candidate material for construction of third generation biosensor and bioelectronics devices.