A simple approach such as the anodic activation of a glassy carbon electrode (GCE) can be used successfully to improve the detection of acetaminophen at very low concentration levels. In this work, we observed that the exposure of a GCE to a high potential (2 V vs Ag/AgCl) for a limited time period (60 s) in the presence of 50 mM borate/phosphate buffer (pH 9) provides a strongly electro-activated surface. The activated surface was characterized by means of several techniques (electrochemistry, EIS, SEM, AFM, mu-Raman, XPS). It appeared that the anodization procedure gave rise to a strong oxygen-based functionalization that did not affect morphologically the electrode surface. The mechanism of interaction of the activated electrode with the analyte was studied by electrochemical methods, then the electrode was applied to the electroanalysis of acetaminophen by differential pulse voltammetry. In this application, the method showed promising analytical performance, detecting nanomolar traces of acetaminophen. (C) 2016 Elsevier Ltd. All rights reserved.