Abstract: The electrochemical behaviour of shikonin was investigated on a glassy carbon electrode in an acetate buffer (pH=3.98) solution using cyclic voltammetry. Shikonin gives a pair of redox peaks by the absorption-controlled process. The experimental conditions were optimized for the determination of shikonin and the square-wave anodic peak currents were linearly related to the shikonin concentrations in the range from 2.08×10^-8 to 1.82×10^-6 mol/L with a correlation coefficient of 0.998. Using the established method without pretreatment and pre-separation, shikonin in herbal drug Gromwell Root was determined with satisfactory result.