High Performance Thin Layer Chromatography (HPTLC) fingerprinting for authentication of monofloral honey

Authenticating the floral origin of honey is a key aspect of quality control, particularly for monofloral honeys, which are derived from a predominant nectar source and commonly yield higher prices than multifloral honeys. While bees can deliberately target and collect pollen, which serves as their main dietary protein source, they may inadvertently also introduce pollen of their floral nectar source into the nectar during collection. This pollen then becomes a natural honey ‘contaminant’. Melissopalynology, the microscopic analysis of pollen in honey, is a widely used method for verifying monofloral honey on the assumption that honey’s predominant pollen grains align with its nectar origin. However, this authentication technique has its limitations. While it is effective when bees forage from a restricted set of floral sources such as in agricultural settings with controlled pollination, it is limited in its ability to authenticate honeys produced from extrafloral nectaries, pollen-free flowers or those produced in biodiverse environments, where a wide variety of pollen may unintentionally ‘contaminate’ the nectar on its collection [ 1 ] [ 2 ] [ 3 ]. Using Jarrah ( Eucalyptus marginata ) honey as a case example, this study highlights the potential of High Performance Thin Layer Chromatography (HPTLC) fingerprinting as an alternative method for monofloral honey authentication in such complex environments. Honey extracts are visualised as sets of HPTLC fingerprints, which capture honey’s typical chemical composition reflective of its nectar origin. The study demonstrates that despite natural variations in composition, the proposed analytical approach delineates typical patterns that can be used to confirm honey’s monofloral status. Publication History Article published online: 05 March 2026 © 2026. Thieme. All rights reserved. Georg Thieme Verlag KG Oswald-Hesse-Straße 50, 70469 Stuttgart, Germany