A multifaceted approach is needed to fully characterise honey composition and evaluate its beneficial effects in biomedical applications. Here we investigated the composition of four samples of Irish honey, marketed as heather honey, and one sample of manuka honey, examined their inhibitory effect on bacterial growth, and developed honey-loaded scaffolds for topical application. The melissopalynological analysis confirmed only one Irish sample as unifloral heather honey. Aroma compounds, previously suggested as markers of botanical origin, were measured in the headspace of Irish heather honey (C9-norisoprenoids with 3,5,5-trimethyl-cyclohexenic structure) and manuka honey (2-methoxyacetophenone and 2’-hydroxyacetophenone) via Gas Chromatography/Mass Spectrometry (GC/MS). Honey’s antibacterial activity against S. aureus and E. coli was evaluated in vitro, with bacterial growth inhibition of 93-100% induced by 50% (w/v) honey solutions in the broth microdilution assay, while the Minimum Bactericidal Concentration (MBC) was established at a honey concentration between >50% and 25% (w/v). An alternative strategy to evaluate honey’s antibacterial properties via GC/MS was proposed and tested with E. coli by monitoring the variation in emission of distinctive volatile metabolites (i.e., indole and linear alcohols) when bacterial growth was inhibited by honey in liquid culture. Honey-based hydrogels were fabricated with the aim to improve honey’s applicability, and their functional performance evaluated regarding swelling degree for absorption of wound exudate and honey release rate. All the hydrogels developed here, regardless of the honey presence, exerted a significant antibacterial effect against S. aureus (but not against E. coli) in vitro, which should therefore be attributed to the intrinsic antibacterial properties of the polymer formulation. It has been demonstrated in this work that Irish heather honey has an antibacterial effect comparable to manuka honey, that its botanical origin can be characterised using GCMS methods, and that it can be successfully incorporated into hydrogels whereupon its release can be manipulated. Together, these elements provide a robust foundation for further investigations into the potential of Irish honey for biomedical use.