Directmethanol fuel cells (DMFCs) offer one of the most promising alternatives for the replacement of fossil fuels. A DMFC that had an active Membrane Electrode Assembly (MEA) area of 45 cm2, a squoval-shaped manifold hole design, and a Pt-Ru/C catalyst combination at the anode was taken for analysis in simulation and real-time experimentation. A mathematical model was developed using dynamic equations of a DMFC. Simulation of a DMFC model using MATLAB software was carried out to identify the most influencing process variables, namely cell temperature, methanol flow rate and methanol concentration during a DMFC operation. Simulation results were recorded and analyzed. It was observed from the results that the cell temperature was the most influencing process variable in the DMFC operation, more so than the methanol flow rate and the methanol concentration. In the DMFC, real-time experimentation was carried out at different cell temperatures to find out the optimum temperature at which maximum power density was obtained. The results obtained in simulation and the experiment were compared and it was concluded that the temperature was the most influencing process variable and 333K was the optimum operating temperature required to achieve the most productive performance in power density of the DMFC.