Screen-printed carbon electrodes (SPCEs) are widely used for the electroanalysis of a plethora of organic and inorganic compounds. These devices offer unique properties to address electroanalytical chemistry challenges and can successfully compete in numerous aspects with conventional carbon-based electrodes. However, heterogeneous kinetics on SPCEs surfaces is comparatively sluggish, which is why the electrochemical activation of inks is sometimes required to improve electron transfer rates and to enhance sensing performance. In this work, SPCEs were subjected to different electrochemical activation methods and the response to H2O2 electroanalysis was used as a testing probe. Changes in topology, surface chemistry and electrochemical behavior to H2O2 oxidation were performed by SEM, XPS, cyclic voltammetry, chronoamperometry and electrochemical impedance spectroscopy. The combination of electrochemical activation methods using H2SO4 and H2O2 proved particularly effective. A reduction in charge transfer resistance, together with functionalization with some carbon-oxygen groups on carbon ink surfaces, were likely responsible for such electrochemical improvement. The use of a two-step protocol with 0.5 M H2SO4 and 10 mM H2O2 under potential cycling conditions was the most effective activation procedure investigated herein, and gave rise to 518-fold higher sensitivity than that obtained for the untreated SPCEs upon H2O2 electrooxidation. The electrochemical behavior of acetaminophen, hydroquinone and dopamine is also shown, as a proof of concept upon the optimum activated SPCEs.