On the basis of the quenching effect of oxygen on the material fluorescence, we developed an analytical method based on time-domain fluorescence lifetime to detect the concentration of dissolved oxygen in water. The fluorescence lifetime is calculated according to two points on the fluorescence quenching curve, and the dissolved oxygen concentration in water is then obtained by the inversion of Stern-Volmer equation. The results showed that under the same dissolved oxygen concentration, the normalized fluorescence quenching curve is not affected by the excitation conditions, such as the intensity of the excitation light and the duration of excitation. The measured fluorescence lifetime of different dissolved oxygen concentrations is affected by the system delay. The theoretical curve of the fluorescence lifetime after compensation is in good agreement with the modified curve with the fitting correlation coefficient of 0.9985. Compared with the HQ30d dissolved oxygen analyzer, the measurement error of the dissolved oxygen in the mass concentration range of 0-20 mg·L -1 is less than 0.5 mg·L -1 while the fitting correlation coefficient reach to 0.9992. These results showed that it could be an effective method for the measurement of dissolved oxygen concentration in water.