In the present investigation, the methanol crossover rate through Nafion®-115 membrane at different temperatures and different concentrations had been investigated in a fuel cell test apparatus by using gas chromatography analysis. The singledirect methanol fuel cell (DMFC) tests were carried out to investigate the effect of the concentration of methanol aqueous solutions and cell temperature on methanol crossover and consequently, on the open circuit voltage and the cell performance of DMFC. It can be found that the methanol crossover rate through Nafion® membrane increases as methanol concentration and temperature increase. It can also be found that methanol crossover presented a negative effect on the open circuit voltage and the single DMFC performance. Single DMFC test results showed that an improved cell performance was obtained as temperature increased although the methanol crossover rate increased with temperature increment.