Mitigation of pesticide dispersion in soil and water is required to protect ecosystem health and the anthropic uses of water bodies. Biochar amendments have been suggested to reduce pesticide dispersion due to their high sorption potentials. Nevertheless, appraisals at different scales have been limited by the costs of pesticide analyses. The aim of this study was to evaluate the potential of two fluorescent tracers, uranine (UR) and sulforhodamine B (SRB), for use as pesticide proxies in the context of biochar amendments used for mitigation purposes. Therefore, we compared the sorption processes of both fluorescent tracers and those of three pesticides, glyphosate, 2,4-D, and difenoconazole for soils; three wood biochars (pine, oak, and beech/charm blend); and soil/biochar mixtures representing agricultural usages. The results showed that the sorption of glyphosate by soil was unaffected by amendment with the tested pine, oak, and wood blend biochars. In contrast, the sorption coefficients of UR, SRB, 2,4-D, and difenoconazole were significantly increased with these biochar amendments. SRB, in particular, exhibited sorption behavior similar to that of the hydrophobic fungicide difenoconazole. This indicates promise for the use of SRB as a proxy for hydrophobic pesticides, in testing biochar amendments.