A native graphene oxide (NGO) prepared by Hummers modified method was used as the template materials for the fabrication of electrochemically oxidative graphene oxide (OGO) onto a glassy carbon electrode (GCE). The fabrication of OGO-GCE persuaded via anodic reversible potentiodynamic of NGO base surface materials that increases the concentration of oxygen functionalities. The development of a new aldehyde/alcoholic functional group on OGO-GCE characterized by XPS analysis is a marked signal for oxygen richness surface which provoked on the cost of sp2 hybridized function. The EIS data underline that OGO-GCE promote the electron transfer kinetics much more than NGO-GCE by 9 times as estimated by the rate constant calculation. The XPS and EIS data highlight the influence of anodic treatment on increasing the interlayer spacing distance between the resultant OGO. Differential pulse voltammetry (DPV) results were evident for a promising efficiency of OGO-GCE on the simultaneous determination of dopamine (DA), ascorbic acid (AA) and uric acid (UA). In addition, the selectivity of the proposed sensor for DA quantification in the presence of high concentrations of AA and UA was achieved successfully with a detection limit (DL3s) approach to 12 nM. The analytical performance of OGO-GCE on serum blood revealed its potential application for trace DA quantification.