BSA, tryptones, and environmental protein-like substances were analyzed in this study. Environmental substances and tryptones had more aromatic groups and were less stable than BSA. Only BSA exhibited significant fluorescence quenching and absorbance changes when interacting with metal ions or humic acids. BSA also tended to aggregate, while tryptones and environmental substances did not. The findings suggest that tryptones are a better model for studying protein-pollutant interactions.