In this paper, multi-walled carbon nanotube/Azure A/gold nanoparticle composites (Nafion/AuNPs/AzA/MWCNTs) were prepared by binding gold nanoparticles to the surfaces of Azure A-coated carbon nanotubes. Nafion/AuNPs/AzA/MWCNTs based electrochemical sensor was fabricated for the simultaneous determination of ascorbic acid, dopamine, uric acid, and tryptophan. Cyclic voltammetry and electrochemical impedance spectroscopy were used to characterize the electrochemical properties of the modified electrodes. The modified electrode showed excellent electrocatalytic activity toward ascorbic acid, dopamine, uric acid, and tryptophan (pH 7.0). The experiment results showed that the linear response range for simultaneous detection of AA, DA, UA and Trp were 300-10,000 mu M, 0.5-50 mu M, 0.5-50 mu M and 1.0-100 mu M, respectively, and the detection limits were 16 mu M, 0.014 mu M, 0.028 mu M and 0.56 mu M (S/N = 3). The proposed method offers promise for simple, rapid, selective and cost-effective analysis of small biomolecules. The procedure was also applied to the determination of tryptophan in spiked milk samples. (C) 2015 Production and hosting by Elsevier B.V. on behalf of King Saud University.