The fraudulent addition of plant oils during the manufacturing of hard cheeses is a real issue for the dairy industry. Considering the importance of monitoring adulterations of genuine cheeses, the potential of fluorescence spectroscopy for the detection of cheese adulteration with plant oils was investigated. Synchronous fluorescence spectra were collected within the range of 240 to 700 nm with different wavelength intervals. The lowest detection limits of adulteration, 3.0 and 4.4%, respectively, were observed for the application of wavelength intervals of 60 and 80 nm. Multiple linear regression models were used to calculate the level of adulteration, with the lowest root mean square error of prediction and root mean square error of cross validation equalling 1.5 and 1.8%, respectively, for the measurement acquired at the wavelength interval of 60 nm. Lower classification errors were obtained for the successive projections algorithm-linear discriminant analysis (SPA–LDA) rather than for the principal component analysis (PCA)–LDA method. The lowest classification error rates equalled 3.8% (∆ λ = 10 and 30 nm) and 0.0% (∆ λ = 60 nm) for the PCA–LDA and SPA–LDA classification methods, respectively. The applied technique is useful for detecting the addition of plant fat to hard cheese.