The present study reveals the utility of high-performance thin-layer chromatographic (HPTLC) fingerprinting of phenolic constituents for the authentication of monofloral honeys. The obtained data enables a more complete assessment of honey quality and the identification of emerging threats to honey quality. The developed procedure facilitates differentiation of varietal honeys and detection of honey adulterations. We used an HPTLC fingerprint analysis to determine the characteristic patterns of different honey types (willow, buckwheat, heather, pine honeydew, and manuka honey). The HPTLC chromatograms were used to determine the differences in the botanical origin of the honey samples on the basis of the band profiles, which are characteristic for each honey type. Identification of 11 polyphenols was performed by comparison of the color and R f of the bands with available standards. Additionally, the results were confirmed by an HPLC analysis.