The fate of fluorescing substances during the recycling of membrane backwash water (MBWW) ahead of coagulation, flocculation and sedimentation (CFS) with ultrafiltration (UF) membranes has been investigated. Bench-scale UF membranes were used to generate MBWW from a CFS-treated surface water containing 21 mg/L dissolved organic carbon (DOC) registering a 0.95 cm–1 UV254 absorbance that had been coagulated with 100 mg/L with polyaluminum chloride. CFS settled water, when processed with UF, produced MBWW containing 9 mg/L DOC registering a 0.25 cm–1 UV254 absorbance. High performance size-exclusion chromatography using UV254 detection demonstrated an analogous UV254 reduction as measured by detector response. However, fluorescence excitation emission spectroscopy revealed that protein-like substances, known to be associated with irreversible fouling, had been concentrated in the MBWW. In order to evaluate recycling operations on overall DOC removal in a CFS-UF process, a blend of 30% MBWW with 70% of raw water was treated, resulting in an overall DOC removal of 73%. However, without MBWW recycle, the CFS-UF process removed less of the influent DOC (63%). This work suggests that MBWW recycle operations should consider possible downstream impacts of concentrated protein-like substances not previously detected, as these substances are suspected to contribute to long-term irreversible UF fouling.