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Front-face fluorescence method has been used to obtain Fluorescence Excitation Emission Matrix (EEM) characteristic of various Italian monocultivar Extra Virgin Olive Oil (EVOO), Mixed EVOO, and lower grade Virgin Olive Oil (VOO). Specific region of EEM has been explored using Principle Component Analysis (PCA). It was found that EEM region of excitation 250-400 and emission 280-620 nm could be used to discriminate each sample type of different cultivar, mixed sample, and virgin olive oil sample. The important fluorescence peaks for the discrimination belong to unique combination of Tocopherols, Tocotrienols, Phenolic compounds, Oxidation products, and Vitamin E. These results show that front-face Fluorescence EEM spectroscopy has a potential to be used for monocultivar EVOO discrimination.