The most common frauds of tuna cans supply chain concern the substitution or mixing of valuable tuna species with cheaper ones, which is strictly forbidden. The objective of the present study was to determine the potential use of front-face fluorescence spectroscopy (FFFS) as a rapid tool to authenticate species in canned tuna: skipjack tuna (Katsuwonus pelamis), yellowfin tuna (Thunnus albacares), Albacore tuna (Thunnus alalunga), and bigeye tuna (Thunnus obesus). Different spectra (tryptophan residues, aromatic amino acids, and nucleic acids (AAA + NA), riboflavin, nicotinamide adenine dinucleotide (NADH), and vitamin A) were recorded on 256 canned tunas, produced at the pilot scale, that were used for the establishment of models. The robustness of the established models was tested on 40 commercial canned tunas. According to the label tunas, the percentage of correct classification reached 75% allowing us to conclude that FFFS may represent a promising tool to be used by both canning industry and governmental control agencies to ascertain correct labeling of canned tuna.