The studies of the interaction between nicotine and DNA on the solid electrode were proposed for the first time. The electrochemical behaviors of nicotine and its interaction with DNA were explored by differential pulse voltammetry (DPV) at the DNA modified glassy carbon electrode (DNA/GCE). In 0.05M Na 2 C 2 O 4 (pH 4.24), nicotine at DNA/GCE showed an irreversible reductive behavior in the range of −1.6 to −1.1V. The presence of DNA led to the decrease in the peak current of nicotine and the negative shift in the potential, indicating that nicotine could interact with DNA by electrostatic mode. The binding ratio between nicotine and DNA was calculated to be 2:1 and the binding constant was 8.31(±0.36)×10 2 . The interaction between nicotine and DNA could be further verified by UV–vis spectroscopy.