In the present work, combination of bi-metallic and tri-metallic Pt, Ir, Sn electro-catalysts was prepared by impregnation reduction method on carbon Vulcan XC-72 to improve upon electro-oxidation of ethanol in direct ethanol fuel cell. The prepared electro-catalysts were characterized by means of scanning electron microscope (SEM), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDX) and X-ray diffraction (XRD) analyses. XRD and TEM analyses reveal that the prepared catalysts are of nano size (6-10 nm) range. It is shown that Pt lattice parameter decreases with the addition of Ir, and increases with the addition of Sn in Pt-Ir-Sn/C catalyst. The electro-catalytic activities characterized by cyclic voltammetry (CV), linear sweep voltammetry (LSV) and chronoamperometry (CA) techniques reveal that the addition of small amount of Ir in Pt-Sn/C electro-catalyst exhibits higher activity towards ethanol oxidation than the Pt-Sn/C (20% Pt and 20% Sn by wt) electro-catalyst. The single direct ethanol fuel cell (DEFq test at 90 degrees C, 1 bar with catalyst loading of 1 mg/cm(2) and 2 M ethanol as anode feed showed an enhancement of catalytic activity in following order: Pt-In-Sn/C (20% Pt, 5% Ir and 15% Sn by wt) > Pt-In-Sn/C (20% Pt, 10% In and 10% Sn by wt) > Pt-Sn/C (20% Pt and 20% Sn by wt) > Pt-In-Sn/C (10% Pt, 15% Ir and 15% Sn by wt) > Pt-Ir/C (20% Pt and 20% Ir by wt) > Pt/C (40% Pt by wt). Pt-In-Sn/C (20% Pt, 5% In and 15% Sn by wt) exhibited highest performance among all the catalysts prepared with power density of 29 mW/cm(2) in DEFC operating at 90 degrees C. Copyright (C) 2011, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.