A New Essential Oil – Agonis fragrans Chemotype Selection and Evaluation A report for the Rural Industries Research and Development Corporation by Christopher J. Robinson August 2006 RIRDC Publication No 06/090 RIRDC Project No GSD -1A © 2006 Rural Industries Research and Development Corporation. All rights reserved. ISBN 1 74151 353 7 ISSN 1440-6845 Chemotype Selection and Evaluation of Agonis fragrans oil Publication No. 06/090 Project No GSD -1A The information contained in this publication is intended for general use to assist public knowledge and discussion and to help improve the development of sustainable industries. The information should not be relied upon for the purpose of a particular matter. Specialist and/or appropriate legal advice should be obtained before any action or decision is taken on the basis of any material in this document. The Commonwealth of Australia, Rural Industries Research and Development Corporation, the authors or contributors do not assume liability of any kind whatsoever resulting from any person's use or reliance upon the content of this document. This publication is copyright. However, RIRDC encourages wide dissemination of its research, providing the Corporation is clearly acknowledged. For any other enquiries concerning reproduction, contact the Publications Manager on phone 02 6272 3186. Researcher Contact Details Chris Robinson c/o 444 Albany Highway ALBANY WA 6330 Phone: 08 98 928486 Fax: 08 98 412707 Email: [email protected] In submitting this report, the researcher has agreed to RIRDC publishing this material in its edited form. RIRDC Contact Details Rural Industries Research and Development Corporation Level 2, Pharmacy Guild House 15 National Circuit BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: 02 6272 4819 Fax: 02 6272 5877 Email: [email protected]. Web : http://www.rirdc.gov.au Published in August 2006 Printed on environmentally friendly paper by Canprint ii Foreword Although Western Australia has one of the most diverse floras in the world (ca. 12,000 taxa), with enormous potential for native plant based industries, to date there has been little research into the development of the essential oils of these species. Over the last decade there has been growing interest in the commercial production and uses of the essential oil of Agonis fragrans, which is a species native to the south coast of WA. This project aimed to investigate chemotype variation of Agonis fragrans across its natural range to ensure that any further commercial development of its essential oil was based upon a suitable chemotype. The project also aimed to confirm antimicrobial status of the oil, identify uses and clarify regulatory processes to increase its commercial application. This research is important to ensure vital background data is available to the emerging industry based upon Agonis fragrans oil. The species itself was only described in 2001 and no scientific data on its chemistry and uses had previously been completed. The report is intended to benefit essential oil producers and users. It is essentially a collation of sub reports on various aspects of research into Agonis fragrans oil. Readers requiring specific information on the research methodologies used are referred to the appendices which contain the complete sub- reports. This project was funded by RIRDC’s core funds which are provided by the government of Australia. Funding support was also provided by the Great Southern Development Commission, Albany. This report is an addition to RIRDC’s diverse range of over 1500 research publications. It forms part of our Essential Oil and Plant Extracts R&D sub-program which aims to support the growth of a profitable and sustainable essential oils and natural plant extracts industry in Australia. Most of our publications are available for viewing, downloading or purchasing online through our website: • downloads at www.rirdc.gov.au/fullreports/index.html • purchases at www.rirdc.gov.au/eshop Peter O’Brien Managing Director Rural Industries Research and Development Corporation iii Acknowledgments The support and funding provided by RIRDC has been critical in facilitating the research contained in this report which will strengthen and enhance the commercial reality of Agonis fragrans oil. RIRDC Research Manger Tony Byrne was always available, responsive and flexible in his dealings with the project partners. Peter Grayling of CALM provided the initial GC analysis of 55 leaf samples which facilitated more targeted sampling for GCMS analysis. The Buxton family of Redmond allowed harvest of their Agonis fragrans plantation for oil production. The Day family of The Paperbark Company provided great enthusiasm and the necessary commercial reason to continue the research into this new oil. Great Southern Development Commission contributed to funding and provided an administrative base through which the partners could develop this project. iv Contents Foreword............................................................................................................................................... iii Acknowledgments................................................................................................................................. iv Contents.................................................................................................................................................. v Executive Summary ............................................................................................................................. vi Introduction ........................................................................................................................................... 1 Project Objectives.................................................................................................................................. 1 Production and Analysis of Oil Samples from Plantations and Natural Populations..................... 2 Comparative Antimicrobial Tests........................................................................................................ 2 Identification of Best or Superior Chemotypes .................................................................................. 2 Identification of Potential Uses ............................................................................................................ 3 Assessment of Immunostimulatory Ability for Aquaculture............................................................. 4 Identification of and Progression Toward Recognition by Regulatory Authority.......................... 4 Determination of Traditional Use ........................................................................................................ 4 Determination of Anti Inflammatory Ability...................................................................................... 5 Discussion............................................................................................................................................... 6 Implications and Recommendations.................................................................................................... 8 Appendices ............................................................................................................................................. 9 Appendix 1: A Report on the Field Investigation of Market Opportunities for Agonis Oil. .............. 9 Appendix 2: Gas Chromatography of 55 Agonis fragrans leaf samples in ethanol .......................... 13 Appendix 3: GCMS analysis and “Comments on the chemistry of the oils”.................................... 19 Appendix 4: Composition of an Anti-microbial Essential Oil from Agonis fragrans....................... 24 Appendix 5: “Antimicrobial activity of Agonis fragrans oil”........................................................... 32 Appendix 6: “Regulatory Overview Agonis Oil” ............................................................................. 44 Appendix 7: Effects of Agonis fragrans oil on mononuclear cell immune responses ...................... 60 v Executive Summary What this report is about This report provides information on research to facilitate realisation of the commercial potential of a new essential oil, Agonis fragrans. Who the report is targeted at The report is particularly targeted at potential investors in the production of agonis oil and potential users of the product. It will also be useful as a first step in investigation of the regulatory requirements for the oil. Background The oil contained in the foliage of Agonis fragrans, an erect shrub native to SW Western Australia, was first recognised as having commercial potential as an essential oil in 1996. Since then, a market investigation confirmed commercial interest, but identified a lack of supply as a limitation to further development. In 2001, The Paperbark Company established a plantation of Agonis fragrans and is now producing, promoting and selling increasing quantities of the oil. Aims This project aimed to investigate chemotype variation of the species across its natural range to ensure that any further commercial development was based upon a suitable chemotype. The project also aimed to confirm antimicrobial status of the oil, identify uses and clarify regulatory processes to increase its commercial application. Methods used The methodology involved production and analysis of oil samples from plantations and across range of natural populations and identification of best or superior chemotypes; comparative antimicrobial