Herein, a portable and cost‐effective electrochemical sbased on Silver/tannic acid/titanium dioxide/glassy carbon electrode (Ag/TA@ TiO2/GCE) was fabricated to determine timolol(TM) assay. The Ag/TA@TiO2/GCE offered an irreversible oxidation peak at +0.99 V, and exhibited an extraordinary electrochemical performance with a wide linear working ranges from 0.01–0.84 and 0.84–49.0 μM and a low detection limit of 5.2 nM. The detection of TM in the presence of interfering agents and real samples was also analyzed. The sensor‘s selectivity was studied by comparing the binding of TM, propranolol, nebivolol, and metoprolol. The developed electrochemical sensing platform could have promising potential for the determination of TM in clinical samples.