Membrane fouling has always been an urgent issue to be tackled in membrane filtration field. The development of new characterization technology of membrane fouling is beneficial for understanding its process and provides reference for its control. This research tried to characterize organic membrane foulants directly through the front-face fluorescence excitation-emission matrix (FF-EEM). Bovine Serum Albumin (BSA), as the standard foulant, was used with Polyvinylidene fluoride (PVDF) membrane for this ultrafiltration experiment. The FF-EEM coupled with parallel factor analysis (PARAFAC) quantified the foulants accumulated on the surface of the membrane during the experiment. The research confirmed the validity of the application of FF-EEM on membrane foulants characterization. The result showed that the BSA deposition rate was relatively slow at the initial stage of the experiment, while as the filtration went on, a filter cake layer was formed on the surface of the membrane and the BSA deposition rate got faster, which led to more filtration resistance. It proved that FF-EEM is accurate and convenient in the characterization of the membrane foulants, and is expected to provide technical support for further research studies and online detection of organic membrane fouling.