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The standardization of the botanical origin of honey reflects the commercial value and quality of honey. Nowadays, most consumers are looking for a unifloral honey. The aim of the present study was to develop a novel method for honey classification using chemometric models based on phenolic compounds analyzed with right angle fluorescence spectroscopy, coupled with stepwise linear discriminant analysis (LDA). The deconstructed spectrum from three-dimensional-emission excitation matrix (3D-EEM) spectra provided a correct classification score of 94.9% calibration and cross-validation at an excitation wavelength (λex) of 330 nm. Subsequently, a score of 81.4% and 79.7%, respectively, at an excitation wavelength (λex) of 360 nm was achieved. Each chemometric model confirmed its power through the external validation with a score of 82.1% for both. Differentiation could be correlated with hydroxycinnamic and hydroxybenzoic acids, which absorb in this region of the spectrum. Fluorescence spectroscopy constitutes a rapid and sensitive technique, which, when combined with the stepwise algorithm and LDA method, can be used as a reliable and predictive authentication tool for honey. This study indicates that the developed methodology is a promising technique for determination of the botanical origin of common Greek honey varieties. Our long-term ambition is to support producers and suppliers to remain in a competitive national and international market.

The Use of Right Angle Fluorescence Spectroscopy to Distinguish the Botanical Origin of Greek Common Honey Varieties