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Abstract: In the present work, using copper as an etching agent, a novel and easy method to prepare porous graphene nanosheet (P-GR) was reported. With a large number of pores, more acidic groups and edge defect sites (EDSs) compared with graphene (GR), the prepared P-GR was used as an electrochemical biosensing platform for the first time. Using voltammetry and amperometry as detection methods, some important electroactive compounds, including dopamine (DA), ascorbic acid (AA), uric acid (UA), epinephrine (EP), guanine (G), adenine (A), xanthine (XA), hypoxanthine (HX), hydrogen peroxide and β-nicotinamide adenine dinucleotide (NADH) were employed to study their electrochemical responses on P-GR modified glassy carbon electrode (P-GR/GCE). The results show that P-GR/GCE exhibits better electrochemical performance to these substances compared with GR modified glassy carbon electrode (GR/GCE) and edge plane pyrolitic graphite electrode (PGE). The remarkable electrochemical advantage of P-GR makes it a promising material in electrochemical biosensing field. [Copyright &y& Elsevier]