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In order to identify possible adulteration of onroad diesel with vegetable oil, fluorescence spectroscopy was used as the analytical technique to differentiate between vegetable oil and biodiesel in diesel blends. Diesel/oil and diesel/biodiesel blends made with different proportions of soy, canola or waste cooking oil were analyzed. The reduced cost of analysis using fluorescence spectroscopy together with the reliability of the results suggest that this technique could be of great use in differentiating between diesel, biodiesel and vegetable oil and could therefore be used for rapid identification or confirmation of adulterated diesel. Furthermore, a compact fluorescence spectrophotometer with an LED excitation source could be used in gas stations or fuel distributors for diesel quality control because of its practicality, low cost and reliability.