Enzymeless hydrogen peroxide (H 2 O 2 ) detection with high sensitivity and excellent selectivity is desirable for clinical diagnosis. Herein, one-dimensional Co 3 O 4 nanowires have been successfully constructed on reduced graphene oxide (rGO) via a simple hydrothermal procedure and subsequent thermal treatment. These Co 3 O 4 nanowires, assembled by small nanoparticles, are interlaced with one another and make a spider web-like structure on rGO. The formation of Co 3 O 4 -rGO hybrids is attributed to the structure-directing and anchoring roles of DDA and GO, respectively. The resulting structure possesses abundant active sites, the oriented transmission of electrons, and unimpeded pathways for matter diffusion, which endows the Co 3 O 4 -rGO hybrids with excellent electrocatalytic performance. As a result, the obtained Co 3 O 4 -rGO hybrids can serve as an efficient electrochemical catalyst for H 2 O 2 oxidation and high sensitivity detection. Under physiological conditions, the oxidation current of H 2 O 2 varies linearly with respect to its concentration from 0.015 to 0.675 mM with a sensitivity of 1.14 mA · mM -1 ·cm -2 and a low detection limit of 2.4 μM. Furthermore, the low potential (-0.19 V) and the good selectivity make Co 3 O 4 -rGO hybrids suitable for monitoring H 2 O 2 generated by liver cancer HepG2 cells. Therefore, it is promising as a non-enzymatic sensor to achieve real-time quantitative detection of H 2 O 2 in biological applications.