The food industry places significant emphasis on ensuring quality and traceability as key components of a healthy diet. To cater to consumer demands, researchers have prioritized the development of analytical techniques that can rapidly and non-invasively provide data on quality parameters. In this study, we propose to use the Delayed Luminescence (DL), an ultra-weak and photo-induced emission of optical photons, as a tool for a rapid evaluation of quality profile associated with fruit ripening, in support of traditional analysis methods. Delayed Luminescence measurements have been performed on cherry tomatoes, with and without the PGI “Pomodoro di Pachino” certification, harvested from two different growing areas of south-eastern Sicily (Italy). Then, DL emissions were correlated with soluble solid content and titratable acidity values, which are known to affect the flavor, the commerciality and the maturity degree of tomato fruits. In addition, we evaluated the changes in the DL parameters with respect to the geographical origin of the cherry tomatoes, with the aim of testing the possibility of applying the technique for identification purposes. The signals of Delayed Luminescence appeared to be good indicators of the macromolecular structure of the biological system, revealing structural changes related to the content of total soluble solids present in the juice of tomatoes analyzed, and they appeared unsuitable for authenticating vegetable crops, since the differences in the photon yields emitted by tomato Lots were not related to territory of origin. Thus, our results suggest that DL can be used as a nondestructive indicator of important parameters linked to tomato fruit quality.