A new method of oxygen transfer efficiency (OTE) measurement for aerobic biochemical wastewater treatment system was introduced. The effluent gas and the influent air around the fan were collected and analyzed with a gas chromatograph. The ratios of the concentration of nitrogen to that of oxygen for both influent and effluent gases were determined. The reduced percentage of the ratio for the influent gases was taken as oxygen transfer efficiency (OTE). The nitrogen gas was used as an inner mark. Using this method, the OTE measurement was not affected by the temperature, water vapor mixture and carbon dioxide produced within the treatment system. The accuracy of the method was analyzed. The standard deviation of the ratios for influent and effluent gases were 0.0006 and 0.0008, respectively. The OTE values of a full-scale aeration tank were detected using this method. The OTE values of the representative spots in the aeration tank varied from 7.3% to 11.5%.