Fluorescently labeled cellulose nanofibrils (CNFs) were used to evaluate CNF leaching from paper according to standard safety assays for food contact materials. Enzymatically pretreated pulp was first labeled with 5-([4,6-Dichlorotriazin-2-yl]amino)fluorescein hydrochloride (DTAF), followed by homogenization to produce fluorescent CNFs of varying degrees of fibrillation. Labeling at the μmolar DTAF/g cellulose level imparted quantitative ppb fluorescence detection of CNFs (LOD of approximately 20 ppb), without significantly altering other material properties, suggesting that DTAF-labeled CNFs are an appropriate mimic for native CNFs and that this approach can be used to detect low CNF concentrations. Cold and hot-water extractions of laboratory papers (100 % CNFs and CNF-fiber blended papers) showed loss values below 3 wt% CNFs, with the finest CNF quality showing the least loss overall and with greater loss experienced under hot water conditions compared with cold water. DTAF-labeled CNFs can be used to address questions related to CNF distribution, localization, and loss.