Antibiotic-resistant bacteria are a major health concern and honey may provide an alternative to anti- biotic use under certain conditions. The antimicrobial action of six Scottish honeys and Manuka Medi- honey® was compared against antibiotic-resistant Acinetobacter calcoaceticus, Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli. Certain Scottish honeys, such as Highland and Portobello honey 2011, were comparable in effectiveness to the established antimicrobial Medihoney®, inhibiting growth to <1 compared to 10 log10 CFU/ml in the control. Heather honey was the next most active while Blossom honeys were less active. Bacteria were inhibited by a sugar-matched control, but to a lesser extent, indicating that antimicrobial activity was associated with non-sugar components, such as poly- phenols. However, total phenol content or antioxidant capacity did not correlate with antimicrobial activity. Liquid chromatography-mass spectrometric analysis revealed that the composition of poly- phenol and non-polyphenol components differed between honeys. In addition, candidate components that could be associated with antimicrobial activity were noted including novel fatty diacid glycoside derivatives not previously identified in honeys.