Light-induced oxidation of semihard cheese has been evaluated by fluorescence spectroscopy. The cheese was packaged in two packaging materials and exposed to different storage conditions, which included light/dark storage, oxygen availability, and storage time (0, 4, 7, 14, 21, 42, 70, or 84 days). Fluorescence excitation-emission matrices (EEM) were analyzed by PARAFAC, which gave an estimation of the pure excitation and emission spectra of the fluorophores and the concentrations of these. This analysis showed the presence of components such as tryptophan, tyrosine, vitamin A, fluorescent oxidation products, and riboflavin. Effects of packaging material, light or dark storage, and storage time were seen. However, there was no effect of the oxygen availability on the fluorescence measurements. The score values obtained by the PARAFAC models and chemical and physical measurements were analyzed together by principal component analysis (PCA). The loadings showed a separation of the variables into three groups; the first group was related to oxidation, the second group was related to the degradation of both riboflavin and vitamin A, and the third group was linked to the protein structure.