<p>This study conducted a comprehensive proteomic and amino acid analysis of five monofloral honeys from the Thrace region: sunflower (<i>Helianthus annuus</i>), oak (<i>Quercus</i>), Christ's thorn (<i>Paliurus spina-christi</i>), bindweed (<i>Convolvulus</i>), and goosefoot (<i>Chenopodium album</i>). The combined use of reverse-phase and normal-phase (HILIC) LC-QTOF methods led to the identification of 39 bee-derived proteins, nine of which (including phospholipase A1 and carboxylic ester hydrolase) were common to all honeys. Plant-derived proteins were found to be most abundant in sunflower honey. NanoDrop quantification revealed protein contents ranging from 0.2% to 0.4%, with Christ's thorn honey exhibiting the highest concentration. Free amino acid profiling via LC–MS/MS identified proline as the predominant amino acid, occurring at very high levels, particularly in Christ's thorn honey (10.184&#xa0;nmol/g). The findings underscore the potential of monofloral honeys for cosmetic applications (e.g., anti-aging formulations) and as dietary supplements. Furthermore, the results suggest that proteomic analysis may serve as an effective method for authenticity testing to combat honey adulteration.</p> Graphical abstract <p></p>