The evolution of the fluorescent compounds during the thermal deterioration of virgin olive oil is not yet well-known. Samples of heated virgin olive oils collected from a fryer every 2 h up to 94 h were analyzed to study their fluorescence spectra as well as the evolution of the concentrations of alpha-tocopherol and individual phenols by high-performance liquid chromatography (HPLC). The regions of the fluorescence spectra of the heated oils, diluted in hexane at 1%, were explained by the content of these compounds with regression coefficients higher than 0.90 (R2 adjusted). The fluorescence intensity recorded at 350 nm and the wavelength of the spectrum maximum in the range of 390-630 nm also allowed for the explanation of the increase of the percentage of polar compounds during the experiment. On the other hand, the spectra of the undiluted heated oils indicated that the maximum of the spectrum of any undiluted oil at 490 nm or beyond is related to a percentage of the polar compounds higher than 25%, which is the maximum percentage accepted for edible oils used in frying processes.