In the present work a sensitive and selective electrochemical sensor was fabricated based on a glassy carbon electrode which has been modified with Pd nanoparticles loaded on Vulcan carbon/conductive polymeric ionic liquid composite nanofibers. The nanostructures were characterized by UV-Vis, FT-IR, FESEM, EDX and XRD techniques. The electrochemical study of the modified electrode, as well as its efficiency for the electrooxidation of tramadol was described in 0.1 M phosphate buffered solution (PBS) (pH 7.0) using cyclic voltammetry, linear sweep voltammetry, chronoamperometry and square wave voltammetry as diagnostic techniques. It has been found that application of the composite nanofibers result in a sensitivity enhancement and a considerable decrease in the anodic overpotential, leading to negative shifts about 200 mV in peak potential. The results exhibit a linear dynamic range from 0.05 mu M to 200 mu M and a detection limit of 0.015 mu M for tramadol. Finally, the modified electrode was used for the determination of tramadol in pharmaceutical and biological samples. (C) 2016 Elsevier B.V. All rights reserved.