A carbon nanosphere electrode (CN) has been fabricated for the electrochemical detection of L-serine and L-phenylalanine. Electrochemical behavior of amino acids was investigated using cyclic voltammetry, differential pulse voltammetry, square wave voltammetry and linear sweep voltammetry. In optimal conditions, the peak current of both amino acids on the CN electrode was found to enhance greatly. A sensitive oxidation peak at 0.085 V was observed in the determination of L-serine and 0.06 V for L-phenylalanine. A linearity between the oxidation peak current and the concentration of both amino acids was obtained in the range of 1-100 lM (R 2 = 0.99783 for L-serine, 0.99618 for L-phenylalanine). The practical application of the carbon nanosphere electrode in the determination of the amino acids possesses high selectivity, sensitivity and stability.