Detection of Adulterated Honey by Fluorescence Excitation-Emission Matrices

Honey is a frequent target of adulteration through inappropriate production practices and origin mislabelling. Current methods for the detection of adulterated honey are time and labor consuming, require highly skilled personnel, and lengthy sample preparation. Fluorescence spectroscopy overcomes such drawbacks, as it is fast and noncontact and requires minimal sample preparation. In this paper, the application of fluorescence spectroscopy coupled with statistical tools for the detection of adulterated honey is demonstrated. For this purpose, fluorescence excitation-emission matrices were measured for 99 samples of different types of natural honey and 15 adulterated honey samples (in 3 technical replicas for each sample). Statistical t-test showed that significant differences between fluorescence of natural and adulterated honey samples exist in 5 spectral regions: (1) excitation: 240–265 nm, emission: 370–495 nm; (2) excitation: 280–320 nm, emission: 390–470 nm; (3) excitation: 260–285 nm, emission: 320–370 nm; (4) excitation: 310–360 nm, emission: 370–470 nm; and (5) excitation: 375–435 nm, emission: 440–520 nm, in which majority of fluorescence comes from the aromatic amino acids, phenolic compounds, and fluorescent Maillard reaction products. Principal component analysis confirmed these findings and showed that 90% of variance in fluorescence is accumulated in the first two principal components, which can be used for the discrimination of fake honey samples. The classification of honey from fluorescence data is demonstrated with a linear discriminant analysis (LDA). When subjected to LDA, total fluorescence intensities of selected spectral regions provided classification of honey (natural or adulterated) with 100% accuracy. In addition, it is demonstrated that intensities of honey emissions in each of these spectral regions may serve as criteria for the discrimination between natural and fake honey.