Thesis Page 1 Ethnopharmacology and chemotaxonomy of essential oil yielding Australian plants Nicholas John Sadgrove Thesis Page 2 Ethnopharmacology and chemotaxonomy of novel essential oil yielding Australian plants Nicholas John Sadgrove ‘As aromatic plants bestow No spicy fragrance while they grow; But crush’d or trodden to the ground, Diffuse their balmy sweets around’ Oliver Goldsmith ‘The captivity, an oratorio (Act 1)’ 1764. ‘Who knows but that England may revive in New South Wales when it has sunk in Europe’ Sir Joseph Banks. ‘On a feeling and sensitive mind a demolished forest impresses unmingled sadness, whereas its primeval grandeur must inspire anyone to immeasurable delight, who is susceptible to the beauties of nature’ Baron Ferdinand Von Müller. ‘Let us regard the forests as an inheritance, given to us by nature, not to be despoiled or devastated, but to be wisely used, reverently honoured and carefully maintained. Let us regard the forests as a gift, entrusted to any of us only for transient care, to be surrendered to posterity as an unimpaired property, increased in riches and augmented in blessings, to pass as a sacred patrimony from generation to generation’ Baron Ferdinand Von Müller. Thesis Page 3 Ethnopharmacology and chemotaxonomy of novel essential oil yielding Australian plants Nicholas Sadgrove Bachelor of Environmental Science with Honours (First Class); Southern Cross University, Lismore, New South Wales, Australia. A thesis submitted for the award of Doctor of Philosophy of the University of New England. February 2014 Supervisor: Graham Lloyd Jones Co‐supervisors; Ben Greatrex and Kenneth Watson. I certify that the substance of this thesis has not already been submitted for any degree and is not currently being submitted for any other degree or qualification. I certify that any help received in preparing this thesis and all sources used have been acknowledged in this thesis. _________ _______________ Date _____________3/2/14 Thesis Page 4 Condensed Table of Contents Acknowledgements. …………………………………………………………………….…………………………………………………..……………..5. Abstract. ………………………………………………………………………………………………………………………………………………………..14. Chapter 1 – Introduction: essential oils and ethnopharmacology in Australia. ……………………………………………….16. Journal Articles Part 1: Eremophila longifolia: ethnopharmacology, essential oil chemotypes and cytogeography. ...................................................................................................................................................55. Chapter 2 - Characterization and bioactivity of essential oils from novel chemotypes of Eremophila longifolia (F. Muell) (Myoporaceae): a highly valued traditional Australian medicine. ……………………………………………………….56. Chapter 3 - A possible role of partially pyrolysed essential oils in Australian Aboriginal traditional ceremonial and medicinal smoking applications of Eremophila longifolia (R. Br.) F. Muell (Scrophulariaceae). ……………….69. Chapter 4 – Isolation and characterisation of (-)-genifuranal; the principal antimicrobial component in traditional smoking applications of Eremophila longifolia (Scrophulariaceae) by Australian Aboriginal peoples. ……………………………………………………………………………………………………………………………………………………………………….80. Chapter 5 – Cytogeography of essential oil chemotypes of Eremophila longifolia (Schrophulariaceae). ……….93. Journal Articles Part 2: Ethnopharmacology of medicinal plants used traditionally by Aboriginal Australians. ………………………………………………………………………………………………………………………………………………………………….…105. Chapter 6 - Chemical and biological characterization of novel essential oils from Eremophila bignoniiflora (F. Muell) (Myoporaceae): a traditional Aboriginal Australian bush medicine. …………………………………………………106. Chapter 7 - Chemical and biological characterisation of solvent extracts and essential oils from leaves and fruit of two Australian species of Pittosporum (Pittosporaceae) used in aboriginal medicinal practice. ………………118. Chapter 8 - Medicinal compounds, chemically and biologically characterised from smoke, solvent and distilled extracts from Australian Callitris endlicheri and C. glaucophylla (Cupressaceae): used traditionally in Aboriginal and colonial pharmacopoeia. …………………………………………………………………………………………….…………………………131. Chapter 9 - Characterisation and bioactivity of essential oils from Geijera parviflora (Rutaceae): a native bush medicine from Australia. ……………………………………………………………………………………………………………………………..146. Chapter 10 - Chemogeography and antimicrobial activity of essential oils from Geijera parviflora and Geijera salicifolia (Rutaceae): two traditional Australian medicinal plants. ………………………………………………………………159. Journal Articles Part 3: Phytochemical and chemotaxonomic investigations. ……………………………………………175. Chapter 11 - Antimicrobial activity of essential oils and solvent extracts from Zieria species (Rutaceae). ……176. Chapter 12 - Dihydrotagetone, an unusual fruity ketone, is found in enantiopure and enantioenriched forms in additional Australian native taxa of Phebalium (Rutaceae: Boronieae). ……………………………………………….………189. Chapter 13 - Composition and antimicrobial activity of the essential oils from the Phebalium squamulosum species complex (Rutaceae) in New South Wales, Australia. ………………………………………………………..………………200. Chapter 14 - Cineole-rich essential oils from Australian Prostanthera Labill. species (Lamiaceae): chemotaxonomy and antimicrobial activity. ……………………………………………………………………………….…….…………212. Part 4: Thesis conclusions and appendices. ……………………………………………………………….……………………….………235. Chapter 15 – Conclusion. …………………………………………………………………………………………….………………………………236. Appendix A – Consolidated list of references. ……………………………………………………………………………….……….……245. Appendix B – Supplementary files chapter 5. ………………………………………………………….……………………………………263. Appendix C – Supplementary files chapter 10. …………………………………………………………….………………………………270. Appendix D – Supplementary files chapter 13. …………………………………………………………………………….………………276. Thesis Page 5 Acknowledgements After more than 500 hours of distillation, and travelling distances in excess of 10,000km searching for plants across the Australian landmass, I have an almost unlimited list of acknowledgements to make. However, at the risk of forgetting those who are going to come into contact with my thesis, I would like to commence acknowledgement within my sphere of supervisors. First and foremost, to my principal supervisor Associate Professor Graham Lloyd Jones, who may possibly have demonstrated the greatest exhibition of resilience and perseverance known to academia, in the careful revision and preparation of this thesis. To my co-supervisors Emeritus Professor Kenneth Watson and Dr Ben William Greatrex, for their ready response and selfless contribution to this thesis. To the CSIRO at Chiswick (Uralla, NSW) and their staff, particularly Dr Nicholas Andronicos, for help with the operation of the flow cytometer and ploidy determination. To Andrew Wallace at the UNE chemistry department, for his technical expertise and assistance, generosity with materials and great sense of humour and patience. For their expertise in plant identification and their generosity in making available the excellent resource of the N.C.W Beadle Herbarium at the University of New England, I would like to thank particularly Mr Ian Telford and Professor Jeremy Bruhl. They have always been prepared to share their great experience and expertise in plant taxonomy, with a novice such as myself. I would like to thank the two great phytochemists Ian Southwell and Erich Lassak for their interest and advice with regard to exploring the various essential oil yielding genera in and around New South Wales. In this context I would also like to thank Dr David Tucker, for his useful chemical advice with regard to nuclear magnetic resonance spectroscopy. For his patience over the email, I thank Robert Chinnock, Australia’s foremost expert on Eremophila. I wish to thank the staff and fellow students in the School of Science and Technology, for their friendship, help and advice. They have put up with many strange smells (not all of them unpleasant I hope) emanating from our laboratory. Finally, I wish to thank my family and friends, for what has been a lot of support in this long and sometimes arduous journey. Individual further acknowledgements have been given at the end of each experimental chapter. Thesis Page 6 List of publications originating from this thesis. Sadgrove, N., Gonçalves-Martins, M., & Jones, G. L. (2014). Chemogeography and antimicrobial activity of essential oils from Geijera parviflora and Geijera salicifolia (Rutaceae): Two traditional Australian medicinal plants. Phytochemistry 104: pp. 60-71 Sadgrove, N., Hitchock, M., Watson, K., & Jones, G. L. (2013). Chemical and biological characterization of novel essential oils from Eremophila bignoniiflora (F. Muell) (Myoporaceae): a traditional Aboriginal Australian bush medicine. Phytotherapy Research 27: pp. 1508-1516 Sadgrove, N., & Jones, G. L. (2013). Antimicrobial activity of essential oils and solvent extracts from Zieria species (Rutaceae). Natural Product Communications 8(6): pp. 741-745 Sadgrove, N., & Jones, G. L. (2013). Characterisation and bioactivity of essential oils from Geijera parviflora (Rutaceae): a native bush medicine from Australia. Natural Product Communications 8(6): pp. 747-751 Sadgrove, N., & Jones, G. L. (2013). Chemical and biological characterisation of solvent extracts and essential oils from leaves and fruit of two Australian species of Pittosporum (Pittosporaceae) used in aboriginal