The Science of Nature

, 102:68 | Cite as

Chemical composition and antimicrobial activity of honeybee (Apis mellifera ligustica) propolis from subtropical eastern Australia

  • Carmelina Flavia Massaro
  • Jack Bruce Simpson
  • Daniel Powell
  • Peter Brooks
Original Paper


Propolis is a material manufactured by bees and contains beeswax, bee salivary secretions and plant resins. Propolis preparations have been used for millennia by humans in food, cosmetics and medicines due to its antibacterial effects. Within the hive, propolis plays an important role in bees’ health, with much of its bioactivity largely dependent on the plant resins the bees select for its production. Few chemical studies are available on the chemistry of propolis produced by Australian honeybees (Apis mellifera, Apidae). This study aimed to determine the chemical composition as well as in vitro antimicrobial effects of propolis harvested from honeybees in subtropical eastern Australia. Honeybee propolis was produced using plastic frames and multiple beehives in two subtropical sites in eastern Australia. Methanolic extracts of propolis were analysed by liquid chromatography with ultraviolet detection and high-resolution mass spectrometry (ultra-high-pressure liquid chromatography (UHPLC)-UV-high-resolution tandem mass spectrometry (HR-MS/MS)) and by gas chromatography mass spectrometry (GC-MS). The resulting chemical data were dereplicated for compound characterisation. The two crude extracts in abs. ethanol were tested in vitro by the agar diffusion and broth dilution methods, using a phenol standard solution as the positive control and abs. ethanol as the negative control. Chemical constituents were identified to be pentacyclic triterpenoids and C-prenylated flavonoids, including Abyssinoflavanone VII, Propolin C and Nymphaeol C. The two propolis crude extracts showed bactericidal effects at the minimal inhibitory concentrations of 0.37–2.04 mg mL−1 against Staphylococcus aureus ATCC 25923. However, the extracts were inactive against Klebsiella pneumoniae ATCC 13883 and Candida albicans ATCC 10231. The antistaphylococcal potential of propolis was discussed, also in relation to honeybees’ health, as it warrants further investigations on the social and individual immunities of Australian honeybees.


Geranyl flavonoids Staphylococcus aureus Klebsiella pneumoniae Mass spectrometry Triterpenoids Candida albicans 



The authors are thankful to Terry Braggins at Analytica Laboratories Ltd, NZ, for HR-MS analyses, to the Craig’s members of ‘Valley Bees’, Gympie, for beekeeping help and to Michael Howes at ‘Tyagarah Apiaries’ and Dieter Horstmann at ‘Eagle Farm’ Tyagarah, NSW, Australia, for supplying honeybee propolis from their beehives.


This study and CFM were financed through the University of the Sunshine Coast studentships.

Authors’ contributions

Conceived and designed the experiments: CFM. Performed the experiments: CFM, JS and DP. Analysed the data: CFM. Contributed reagents/materials: PB. Wrote the manuscript: CFM, JS and PB.

Compliance with ethical standards

The authors declare that the experiments described in this article comply with the current laws for the conduct of scientific research in Australia.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

114_2015_1318_MOESM1_ESM.doc (3.9 mb)
ESM 1 (DOC 3966 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Carmelina Flavia Massaro
    • 1
  • Jack Bruce Simpson
    • 2
  • Daniel Powell
    • 1
  • Peter Brooks
    • 1
  1. 1.Faculty of Science, Health, Education and EngineeringUniversity of the Sunshine CoastMaroochydoreAustralia
  2. 2.Research School of BiologyAustralian National UniversityCanberraAustralia

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