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A new approach for the botanical origin determination of monofloral bee honey is developed. The methodology combines mineral content and physicochemical parameters determination with intelligent statistics such as self-organizing maps (SOMs). A total of 62 monofloral bee honey samples were analysed, including 31 linden, 14 rapeseed, 13 sunflower, and 4 acacia. All of them were harvested in 2018 and 2019 from trusted beekeepers, after confirming their botanical origin, using melissopalynological analysis. Nine physicochemical parameters were determined, including colour, water content, pH, electrical conductivity, hydroxymethylfurfural content, diastase activity, specific optical rotation, invertase activity, and proline. The content of thirty chemical elements (Ag, Al, As, B, Ba, Bi, Ca, Cd, Co, Cr, Cs, Cu, Fe, Ga, In, K, Li, Mg, Mn, Na, Ni, P, Pb, Rb, S, Se, Sr, Te, V, and Zn) was measured using ICP-OES, ICP-MS, and FAAS as instrumental techniques. The visualisation of the SOMs shows an excellent separation of honey samples in five well-defined clusters—linden, rapeseed, acacia, sunflower, and polyfloral honey—using the following set of 16 descriptors: diastase activity, hydroxymethylfurfural content, invertase activity, pH, specific optical rotation, water content, Al, B, Cr, Cs, K, Na, Ni, Rb, V, and Zn.

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A New Approach for Determination of the Botanical Origin of Monofloral Bee Honey, Combining Mineral Content, Physicochemical Parameters, and Self-Organizing Maps