Carbamazepine is a popular drug that has been detected in natural environments, but little is known about its biogeochemical cycling, influencing factors, and eco-environmental effects in aquatic ecosystems. Interaction between carbamazepine and humic substances, including fulvic and humic acids, was studied using three-dimensional excitation-emission matrix fluorescence spectroscopy and synchronous-scan fluorescence spectroscopy. The intrinsic fluorescence of humic substances was quenched on the addition of carbamazepine, and static quenching was the primary mechanism. The binding parameters on their interaction, including the conditional binding constants (log K) and binding capacities (C(L)), were estimated by the Ryan-Weber nonlinear theory equation. Log K ranged from 3.41 to 5.04 L/mol at 25 degrees C and pH 7.0. The influence of pH on the complexation and the competition between carbamazepine and Cu(II) for fluorescence-binding sites also were discussed. The present results would be helpful in understanding the fate and biogeochemical cycling of other pharmaceuticals and personal care products in aquatic ecosystems.