The cyclic voltammetry and spectroscopy have been proved to be powerful electrochemical means for the elucidation of interaction betweens Deoxyribonucleic acid (DNA) and chemical anti-carcinogens. With the aim to elucidate the interaction between DNA and anti-carcinogenic 5-fluorouracil derivatives using the instruments and techniques above, the electrochemical behavior of (S)-2-(5-Fluorouracil-1-Acetyl) amido -1,4 -succinic acid (5FU-ASP) and the electrostatic interaction of 5FU-ASP with DNA using Fe(CN)(6)(3-/4-)as electroactive indicator of dominance were investigated in this study. The binding equilibrium constant of interaction is estimated to be 1.1x10(3)xM(-1) and DNA modified gold electrodes were prepared by the dry adsorptive method. Based on the electrochemical and ultraviolet-visible spectrum data, the interaction mode of DNA with 5FU-ASP was further explored, and the results serve as good reference for studies on 5FU-ASP targeting DNA strands or base pairs in the natural environment of living cells in vivo.