A sensitive electrochemical method for lomefloxacin (LMF) determination has been developed by using poly-melamine layer modified glassy carbon electrode [p-(melamine)/GCE]. The surface morphology of the modified sensor was characterized by Horizontal Attenuated Total Reflectance-infrared spectroscopy (HATR-IR), Field emission scanning electron microscopy (FE-SEM) and electrochemical impedance spectroscopy (EIS). The electrochemical measurements were carried out using square wave voltammetry and cyclic voltammetry. The polymer modified electrode showed an excellent electrocatalytic activity toward the electrochemical oxidation of LMF with a well-defined voltammetric peak at similar to 980 mV. The peak current of LMF was found to be linear in the range of 0.1-500 mu M and a detection limit and sensitivity of 16 nM and 0.0346 mu A mu M-1, respectively, were observed. The modified electrode shows good selectivity toward the specific recognition of LMF in the presence of common biological interferents. The analytical applicability of the proposed method was successfully demonstrated in different pharmaceutical formulations, human urine and serum samples. The results obtained from the voltammetry were also validated with HPLC. (C) 2014 The Electrochemical Society. All rights reserved.