Amperometric detection of nicotine (NIC) was carried out on a titanium dioxide (TiO(2))/poly(3,4-ethylenedioxythiophene) (PEDOT)-modified electrode by a molecular imprinting technique. In order to improve the conductivity of the substrate, PEDOT was coated onto the sintered electrode by in situ electrochemical polymerization of the monomer. The sensing potential of the NIC-imprinted TiO(2) electrode (ITO/TiO(2)[NIC]/PEDOT) in a phosphate-buffered saline (PBS) solution (pH 7.4) containing 0.1M KCl was determined to be 0.88 V (vs. Ag/AgCl/saturated KCl). The linear detection range for NIC oxidation on the so-called ITO/TiO(2)[NIC]/PEDOT electrode was 0-5mM, with a sensitivity and limit of detection of 31.35 microA mM(-1)cm(-2) and 4.9 microM, respectively. When comparing with the performance of the non-imprinted one, the sensitivity ratio was about 1.24. The sensitivity enhancement was attributed to the increase in the electroactive area of the imprinted electrode. The at-rest stability of the ITO/TiO(2)[NIC]/PEDOT electrode was tested over a period of 3 days. The current response remained about 85% of its initial value at the end of 2 days. The ITO/TiO(2)[NIC]/PEDOT electrode showed reasonably good selectivity in distinguishing NIC from its major interferent, (-)-cotinine (COT). Moreover, scanning electrochemical microscopy (SECM) was employed to elucidate the surface morphology of the imprinted and non-imprinted electrodes using Fe(CN)(6)(3-)/Fe(CN)(6)(4-) as a redox probe on a platinum tip. The imprinted electrode was further characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR).