Detection of various physical and chemical indicators in the national standard of extra virgin olive oil necessitates the use of expensive physical and chemical analysis instruments and related professionals, which limits the supervision of olive oil market. In this study, a novel olive oil quality analysis algorithm based on the regional integral ratio of the three-dimensional fluorescence characteristic peaks is proposed, in which the characteristic fluorescence peaks of extra virgin olive oil, refined olive oil, and other edible vegetable oil are extracted using their three-dimensional fluorescence spectra. Ten samples were prepared, including four brands of extra virgin olive oils, refined olive oil, and adulterated olive oils, which are extra virgin olive oil doped with 10%-50% refined oil. The quality factor’s dynamic range for olive oil is 37 dB. It can also effectively identify various proportions of doped samples, brand extra virgin olive oil, and fake extra virgin olive oil. The experimental results show that the quality factor of the proposed analysis algorithm outperforms the samples’ three traditional physical and chemical indicators. Since the approach requires less fluorescence spectroscopy data than other fluorescence approaches, it can provide a new theoretical basis for the low-cost and rapid identification instrument of olive oil quality.