This work is focused on the application and performance of a high temperature PBI-based direct ethanol fuel cell, studying the influence of some operating variables such as the temperature, ethanol concentration and oxygen partial pressure. An increase in the temperature resulted in an improvement of the cell performance due to the enhanced electrodic kinetic and electrolyte conductivity. An ethanol/water weight ratio between 0.25 and 0.5 was found to be suitable for providing both enough water and fuel availability to make the ethanol oxidation possible. Measurements of the ethanol crossover at different temperatures and concentrations were carried out. An intermittent lifetime test showed that the cell, after several hours, was able to reach stability. Moreover, its performance was completely reversible with no perceptible losses for 7 days. Finally, tests using bio-ethanol as fuel were performed, with no significant power losses. This final feature is of special interest from a practical 'green' point of view. © 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.