Estuary is a complex region due to the mixing of fresh and sea water as well as the mixing of terrestrial and marine dissolved organic matters (DOM), so that the sorption behavior between hydrophobic organic compounds and dissolved organic matters is very complicated to estimate. By applying fluorescence quenching method, we investigated the influence of the mixing of binary dissolved organic matters from different sources on the binding constant (Kdoc) of pyrene and DOM. Results show that the kinetic of mixing of binary DOMs influences the binding constant, that means if there weren¡¦t enough standing time for mixing of binary DOMs, it would cause some bias in estimating the binding constants of pyrene and DOMs mixture. In this study, we also found that there exists a linear relationship on the binding constants between pyrene and DOMs mixture against mixing ratios. In addition, the mixing mechanism of SRNOM and LHA was dominated by diffusion process from the results of initial mixing efficiency. However, the diffusion process might not be the only mechanism of the mixing reaction of SHHA and LHA, other factors should be considered. Further investigations should be done to find out the dominating factors on the mixing of SHHA and LHA system in the future.