essential oils & plant extracts - agrifutures australia contents 2.4 essential oils research in...

RIRDC Completed Projects in 2001-2002and Research in Progress as at June 2002

Sub-Program 2.4

Essential Oils & Plant Extracts

August 2002 RIRDC Publication No 02/065

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© 2002 Rural Industries Research and Development Corporation. All rights reserved. ISBN 0 642 8464 8 ISSN 1440-6845 "RIRDC Completed Projects in 2001-2002 Research in Progress as at June 2002- 2.4 Essential Oils and Plant Extracts" Publication No: 02/065 The views expressed and the conclusions reached in this publication are those of the author and not necessarily those of persons consulted. RIRDC shall not be responsible in any way whatsoever to any person who relies in whole or in part on the contents of this report. 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.

RIRDC Essential Oils and Plant Extracts Mr Tony Byrne RIRDC Level 1, AMA House 42 Macquarie Street BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: 02 6272 5472 Fax: 02 6272 5877 Email: [email protected] RIRDC Publications Rural Industries Research and Development Corporation Level 1, AMA House 42 Macquarie Street BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: 02 6272 3186 Fax: 02 6272 5877 Email: [email protected] Website: http://www.rirdc.gov.au Published in August 2002 Printed on environmentally friendly paper by Canprint

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Foreword This year RIRDC has produced Research in Progress, June 2002, which contains short summaries of continuing projects as well as those that were completed during 2001-2002 for all of the Corporation’s 20 program areas. The complete report on all the programs is only available in electronic format on our website at http://www.rirdc.gov.au The following report is a hardcopy extract, covering Sub-program 2.4. It contains all entries from continuing Essential Oils and Plant extracts research projects funded by RIRDC. This program aims to support the growth of a profitable and sustainable essential oils and natural plant extracts industry in Australia. This report is the newest addition to our extensive catalogue of almost 800 research reports, videos and CD-Roms of project supported by RIRDC. Please contact us for the latest publications catalogue of view it on our website: • Downloads at www.rirdc.gov.au/reports/Index.html • Purchases at www.rirdc.gov.au/eshop Simon Hearn Managing Director Rural Industries Research and Development Corporation

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Contents

2.4 ESSENTIAL OILS RESEARCH IN PROGRESS

PROJECT

No PROJECT TITLE RESEARCHER PHONE ORGANISATION PAGE

No

Development and Improvement of Products DAQ-269A The control of chalkbrood disease with

natural products (eg essential oils) Dr. Craig Davis (07) 3406 8611 Dept of Primary Industries

(Qld) 1

DAV-168A Kesom oil a new essential oil for flavours Mr. Fred Bienvenu (03) 5731 1222 Dept of Natural Resources & Environment (Vic)

2

MRA-1A Efficient sourcing and fractionation of sesquiterpenoid alcohols from Australian sandalwood

Ms. Valerie Gearon (08) 9841 7788 Mt Romance Australia Pty Ltd 3

UNC-6A Generation of high quality Australian Skullcap products

Prof. Ron Wills (02) 4348 4140 The University of Newcastle 4

UNC-11A Generation of high quality Australian valerian products


UT-27A Optimisation of parsley seed oil production Prof. Robert Menary

(03) 6226 2723 University of Tasmania 6

UT-34A Preliminary evaluation of some medicinal herbs

Dr. Rowland Laurence


UT-35A Development of a celery oil and extract industry in Tasmania

Prof. Robert Menary


Improved Production Systems DAV-190A Implementing IPM in Australian peppermint

crops Mr. Fred Bienvenu (03) 5731 1222 Dept of Natural Resources &

Environment (Vic) 9

UNC-13A Optimisation of polysaccharides in processed echinacea purpurea products

Dr. Douglas Stuart (02) 4548 4124 The University of Newcastle 10

UT-33A Commercial Borage Production for Oil and Gamma-linolenic Acid

Dr. Rowland Laurence


Regulatory Approvals UT-36A Determination of pesticide minimum

residue limits in essential oils Prof. Robert Menary


Industry Development PSE-2A Production of a newsletter of the Essential

Oil Producers Association of Australia Dr. Erich Lassak (02) 9875 1894 Phytochemical Services 13

2.4 ESSENTIAL OILS COMPLETED PROJECTS

PROJECT

No PROJECT TITLE RESEARCHER PHONE ORGANISATION PAGE

No

Development and Improvement of Products DAQ-281A Growing milkweed, a plant with prospective

anti-cancer properties Dr. Craig Davis (07) 3406 8611 Dept of Primary Industries

(Qld) 14

UT-28A Chamomile varieties for essential oil and dry flower production

Prof. Robert Menary


Improved Production Systems DAV-178A Potential for IPM in Peppermint growing in

South East Australia Mr. Fred Bienvenu (03) 5731 1222 Dept of Natural Resources &

Environment (Vic) 17

SAG-1A Methodology and viability of re-establishing commercial boronia plantations

Dr. Doris Blaesing (03) 6427 0800 Serve-Ag Pty Ltd 18

UNC-8A Production of high quality Australian ginseng


Industry Development SAG-2A Review of lavender research and

development Dr. Lee Peterson (03) 6233 5522 Serve-Ag Pty Ltd 21

1

Research in Progress Project Title

The control of chalkbrood disease with natural products (eg essential oils)

RIRDC Project No:

DAQ-269A

Start Date: 01/07/00 Finish Date: 30/04/02 Researcher: Dr. Craig Davis Organisation: Department of Primary Industries (Qld)

Centre for Food Technology 19 Hercules Street HAMILTON QLD 4007

Phone: (07) 3406 8611 Fax: (07) 3406 8677 Email: [email protected] Objectives

• To conduct a laboratory study on the efficacy of natural products against

Chalkbrood bacteria. • To develop an appropriate field treatment plan to evaluate the products

identified above.

Current Progress

The honeybee, Apis mellifera, provides essential pollination for approximately 1 billion dollars worth of agricultural crops in Australia each year. Bee diseases represent a significant problem in the Australian beekeeping industry, causing major economic loss to apiarists. Chalkbrood is an infectious disease of honeybee larvae caused by the spore-forming fungus (Ascosphaera apis). Although chalkbrood does not affect the quality of honey and is of no concern to consumers, it can reduce honey production and cause mortality in excess of 50% in managed bee populations. In 1993, when chalkbrood was first identified in Australia, the New Zealand beekeeping industry was suggesting that chalkbrood was responsible for a 5% reduction in honey production. This figure has been used for the Australian situation for some years. It is difficult to accurately assess the impact of the disease since every State of Australia is infected with the disease. The susceptibility of the chalkbrood fungi to a range of treatments has been examined in the laboratory. The treatments which have been studied include a selection of natural products (eg essential oils), fatty acid derivatives (eg glycerol monolaurate), food acids (eg sorbic, formic and acetic), and some natural extracts (eg from banana, garlic and mango), along with antimicrobial (Leptospermum) honey, natural antimicrobial peptides, and salt. More than 40 essential oils have been assessed including native (tea tree oil, lemon myrtle oil and Manuka oil) and exotic spice oils (like sage, rosemary and clove oil). The current knowledge in this area has been reviewed and will be presented with the final report, along with an outline for a field trial for the evaluation of the most active and appropriate anti-microbiological agents in experimentally-infected hives. If these trials are subsequently undertaken, the success of treatments will be assessed by regular examination for disease development and culture of adult bee samples for chalkbrood. This work has the potential to improve the economic viability of the Australian apiarist while producing an effective and environmentally-sound treatment regime.

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Project Title

Kesom oil a new essential oil for flavours

RIRDC Project No:

DAV-168A

Start Date: 29/07/99 Finish Date: 30/10/02 Researcher: Mr. Fred Bienvenu Organisation: Department of Natural Resources & Environment

NRE Ovens PO Box 235 MYRTLEFORD VIC 3737


To facilitate development of a commercially viable industry based on production of kesom by:

• Development of an effective weed control protocol. • Establishing the effects on oil yield and composition of interactions

between rate/time of nitrogen application and the time /number of harvests.

• Evaluating market opportunities for a range of kesom oils produced under commercial conditions.

• Establishment of likely costs of production of a commercial scale kesom oil production system.

• Provision of a detailed report documenting requirements for the sustainable production and marketing of kesom oil.

Current Progress

Kesom oil is extracted from Persicaria odorata, a Polygonaceae plant very rich in C10 & C12 aldehydes and C10 & C12 alcohols. These naturally produced flavour compounds are sought after in the food and fragrance industries. This is a collaborative project involving RIRDC, a commercial essential oil producer, the flavour and fragrance industry and DNRE to develop the protocols required to produce kesom oil on a commercial basis. The primary aim of the project is to establish the potential to produce kesom oil as a viable essential oil crop. This will be achieved using data developed by NRE agronomic studies now approaching completion at NRE Ovens and market evaluation and development by industry. Larger samples of kesom oil have been produced in Corryong and NRE Ovens in 2001and again in 2002. These samples will enable more product evaluation and attract further development by the flavour and fragrance industry. Weed control protocols have been developed and refined for use as both pre and post plant applications. The kesom plant while used widely in Asia and increasingly in Australia, as a fresh and dried herb/condiment has not yet been developed as a source of a discrete essential oil.

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Project Title

Efficient sourcing and fractionation of sesquiterpenoid alcohols from Australian sandalwood

RIRDC Project No:

MRA-1A

Start Date: 01/07/00 Finish Date: 31/07/03 Researcher: Ms. Valerie Gearon Organisation: Mt Romance Australia Pty Ltd

Lot 2, Down Road Miranmbeena Park ALBANY WA 6330


• The primary objective is the development of methods of the efficient

separation of individual sesquiterpenoid alchohols (or enriched concentrates) from the fragrant wood oil of Western Australia sandalwood. Santalum spicatum (R.Br) A. DC. The pure alcohols or their enriched concentrates will be used in the formulation of value added products such as pharmaceuticals (anti-inflammatory and bacterial agents) cosmetics/toiletries and fine perfumes.

• A secondary, but nonetheless indispensible, objective for the ultimate commercial success of this project is the identification of all parameters influencing the wood oil composition and oil yield.

Current Progress

Mt Romance (MRA) is currently scaling up distillation processes which enable a greater yield of high quality sesquiterpenoid alcohols to be produced. This involves reducing the amount of non – volatile constituents present in the oil, thereby increasing concentrations of the biologically active alcohols. As a result of these technological advances, the company is consistently producing a high odour and therapeutic quality product. A specification for oil of Santalum spicatum has been submitted to Standards Australia. This standard is based on the statistical analysis of a large number of individual distillates. Enriched concentrates of the sesquiterpenoid constituents of sandalwood oil have been assayed to quantify their therapeutic effects. MRA has produced 2 monographs based upon the potency of fractions of oil which contain higher levels of epi – α – bisabolol and E, E farnesol, both established therapeutic chemicals not present in Indian sandalwood oil. This potentially increases the value of the oil by increasing access to the pharmaceutical market. Elucidation of activity in all constituents will enhance this value -adding. Identification of parameters affecting oil composition and yield is proceeding, with cooperation from the Western Australian Government enabling the compilation of chemical profiles according to geographical location of wood. This database is very revealing, and shows a diverse variation in composition which is extremely specific for each location. The information will be of great benefit to the whole sandalwood industry, including farmers considering plantations, researchers identifying pharmaceutical compounds and oil (MRA) and incense manufacturers.

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Project Title Production of high quality Australian skullcap products

RIRDC Project No:

UNC-6A

Start Date: 30/06/98 Finish Date: 30/09/02 Researcher: Prof Ron Wills Organisation: The University of Newcastle

Centre for Food Industry Research and Development PO Box 127 OURIMBAH NSW 2258


• The project seeks to generate technical information to enable skullcap

growers and processors to optimise their growing and harvesting practices and postharvest procedures to maximise quality in all end products that may be marketed domestically or for export.

Current Progress

The effect of post-harvest storage of dried skullcap powder on the flavone active constituents were investigated. Samples were placed in high and low humidity environments at 5°C, 20°C and 30°C in a dark chamber and in a chamber at 20°C under incandescent light. The results revealed there was no significant difference in flavones between all samples except at 5ºC in high humidity which decreased markedly. A common processing method for the manufacture of skullcap products in Australia is by obtaining an extract from dried plant material with a solvent containing ethanol and water. A preliminary study was conducted to determine the relative effectiveness of two methods of physical extraction (sonication and low velocity stirring). The results showed no significant difference between either method. However, the effect of ethanol/water mixtures as the extracting solvent had a significant effect on the extraction of flavones. The maximum amount extracted from aerial material was 72% which was obtained using 60% and 40% ethanol. The extraction level decreased with increasing alcohol content and only 0.5% was extracted with 100% ethanol. The sum of the extracted and residual levels was not consistent for all solvent mixtures, indicating that there was varying degrees of degradation of total flavones during processing.

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Project Title

Generation of high quality Australian valerian products

RIRDC Project No:

UNC-11A

Start Date: 30/07/99 Finish Date: 30/07/02 Researcher: Prof. Ron Wills Organisation: The University of Newcastle

PO Box 127 OURIMBAH NSW 2258


• The project will generate information to enable valerian growers and

processors to optimise growing and harvesting practices and postharvest procedures to maximise quality in the end product. The derived systems will be trialed and optimised with industry groups under commercial conditions.

Current Progress

Ease of washing, drying time and active constituents was examined for the whole root and for rootlets and crown plant sections. There was no effect on washing time, two phase drying was more efficient with the rootlets drying much faster than crowns and whole plants and total valerenic acid and essential oil content was higher in plants that had their rootlets removed than plants left whole, probably because of the over-drying of rootlets in whole plants. Plants were dried at 15 - 70°C to determine the effect of drying temperature on the active constituents. For total valerenic acids there was a significant decrease in the content from 15°C to 40°C, no difference between 40 and 60°C, and a decrease at 70°C. Oil content was reduced and valepotriate content was not affected by increasing drying temperature. The long term storage of powdered valerian root over 18 months showed increase loss of all active constituents in higher humidity, temperature and longer time.

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Project Title

Optimisation of parsley seed oil production

RIRDC Project No:

UT-27A

Start Date: 30/07/99 Finish Date: 01/11/02 Researcher: Prof. Robert Menary Organisation: University of Tasmania

School of Agricultural Science GPO Box 252-54 HOBART TAS 7001


• The location and high apiole parsley seed germplasm will enable the

establishment of a new product line within the essential oil industry in Tasmania. The concurrent development of an efficient extraction protocol for the oil will enhance the returns to the industry.

Current Progress

Two semi-commercial field trials of Plain Leaf parsley were grown concurrently this season. One was a second year trial of the variety "Napoli" and the second a first year planting of the Yates Pty Ltd variety "Dark Green". Both were serially harvested throughout the growing season. Laboratory steam distillations of these samples showed that Dark Green produces higher quality oil than Napoli. The trial demonstrated that both varieties must be maintained through to seed maturity in order to obtain the best oil quality. This poses some difficulties for growers as the seed may shed if the crop is harvested too late. Harvest must be carefully timed and some modifications to the current harvesting techniques will be required. Parsley oil has a specific gravity very similar to that of water and separation of the oil and water as it leaves the condenser is very difficult in the current commercial separator. Solvent partitioning of the wastewater leaving the separator has shown that significant quantities of oil are being lost. This is partly overcome by using two separators in series. A review of the success of a further technique involving salting the water of the second separator is underway. The new variety is currently being incorporated into local production and the results of the review of the extraction technology will be available for commercial growers in the coming season.

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Project Title

Preliminary evaluation of some medicinal herbs

RIRDC Project No:

UT-34A

Start Date: 01/01/01 Finish Date: 31/12/03 Researcher: Dr. Rowland Laurence Organisation: University of Tasmania

GPO Box 447 BURNIE TAS 7320


• To pursue the development and commercialisation of one or more of the

plants investigated. Decisions on development will be based upon the data obtained from the project, including additional market information and other investment criteria.

• The study will deliver comparative data to the stakeholders on the growth and marketable yield of species/genotypes sourced from different parts of the world. It will provide market information on prices and demand for the four herb plants under investigation, additional to that back-grounded in the Keane Report and in the current research proposal.

• Production agronomy aspects will be supported by initial laboratory assessments of extraction procedures. These data will be adequate to allow decisions by the company on further investments in these medicinal herb crops with regard to their suitability for production in the cool, temperate regions of Australia.

• Activities will include the field demonstration to growers, progressively of the crops and of the results obtained.

Current Progress

The project began in 2001 with the accession of lines of astragalus, bilberry, evening primrose and stevia. While evening primrose was originally accessed, discussions with buyer and consultant contacts in Europe in July 2001 convinced the project team that feverfew, Tanacetum parthenium, would be a preferable alternative subject of investigation. Thus, with RIRDC approval, feverfew has been substituted into the project. Thirteen lines of these four species have be acquired to date and planted in small quantity with mixed success. Astragalus lines germinated and grew readily in pots and seedlings have recently been transferred to field plots. One line of feverfew was direct-seeded into the field by the stakeholder, Botanical Resources Australia, and has grown well from a poorly germinated stand. Attempts to access vegetative material of bilberry have failed and only seed material can practically be brought to Tasmania due to quarantine restriction. Three lines of seed have been acquired. Two of these have been germinated in laboratory and greenhouse but seedling growth so far has been poor under these conditions. Five lines of stevia have been acquired, including one from Professor Midmore’s team at CQU, which also has recently received support from RIRDC for stevia research and with whom the current project will collaborate. Initial germination of seed was poor but improved with selection. Seedling growth has also so far been poor under greenhouse conditions. Some established plants have recently been acquired from a commercial nursery in Tasmania and planted in the field. Initial indications are than the species may be readily propagated by cuttings. Access to lines from China and elsewhere which are high in active constituents will be important in the development of a local industry.

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Project Title

Development of a celery oil and extract industry in Tasmania

RIRDC Project No:

UT-35A




• The inclusion of celery (Apium graveolens L.) oil and extract products in

the essential oils industry within Tasmania will increase the diversity of crops available to growers and provide a buffer against price fluctuations in local and international markets. Any refinement of current extraction processes, as a result of the development of the new products has the potential to benefit the entire industry.

Current Progress

Seed of seven celery varieties was solvent extracted and the composition of the extracts examined using gas chromatography. GC/MS is currently being used to identify the various major volatile constituents of these extracts. Protocols are being developed which can later be used for quality assurance of the commercial products. Seedlings of six of these varieties are being tested in a field trial and will be serially harvested through the coming flowering season. This trial will establish the changes in oil composition through the growing season and develop test products including leaf, herb and seed oils for market assessment. Further potential products are a seed oleoresin and a root tincture.

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Project Title

Implementing IPM in Australian Peppermint crops

RIRDC Project No:

DAV-190A

Start Date: 1/09/01 Finish Date: 31/08/04 Researcher: Mr. Fred Bienvenu Organisation: Department of Natural Resources & Environment

NRE Ovens PO Box 235 MYRTLEFORD VIC 3737


• Identification and integration of chemical and sustainable bio-control

practices that will maximise yield of high quality peppermint oil. • Development of data required to support registration (minor or full) of

predator “friendly” pesticides. • Establishment of an industry reference group to facilitate and implement the

use of a practical IPM program.

Current Progress

Following the pilot studies in DAV 178A, which examined the potential for IPM in peppermint, this project seeks, among broader objectives, to implement sustainable use and maintenance of predator mites (Phytoseiulus persimilis) to control two-spotted-mites (TSM) (Tetranychus urticae). The 2001-2002 summer was much cooler than typically and consequently the two-spotted mites were not active in the field until much later than expected. For the same reasons the commercially produced predator mites used for dissemination were not available until late December. As the weather warmed in the peppermint growing areas, numbers of TSM increased rapidly. Quickly the number suggested in overseas studies as the threshold of permanent leaf damage resulting in oil yield reduction was surpassed. Once the predators were released into “hot spots” in the numbers of TSM, the level of infestation was quickly reduced to a point below which no economic damage can be expected. The TSM population fell by over 97% within four weeks after the introduction of the predators. Further observations will be performed throughout the project to assess the over-wintering ability of the predator. Observation and understanding of the wider pest spectrum in the peppermint fields has been a positive side benefit of this study.

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Project Title

Optimisation of polysaccharides in processed echinacea purpurea products

RIRDC Project No:

UNC-13A

Start Date: 01/01/01 Finish Date: 30/12/02 Researcher: Dr. Douglas Stuart Organisation: The University of Newcastle

Centre for Food Industry Research and Development PO Box 127 Brush Road OURIMBAH NSW 2258


• To determine if echinacea extracts based on polysaccharide content could

be grown and processed in Australia to eliminate the need for imported equivalents and develop a potential export market. The success of the project will encourage alternative crops of echinacea and other medicinal herbs within the primary producing and manufacturing sector.

Current Progress

Analysis of the polysaccharides within each section of the plant during growth is showing qualitative and quantitative differences seen in planting location, plant section and growing stage. The results indicate that the root produces a distinctive polysaccharide identity to that in the leaf, stem and flower which are of similar structure. Quantification is being performed with reference to dextran standards of known molecular weight, however, these may be adjusted to directly reference the polysaccharides if current purification through the use of a preparative HPLC is successful. In addition, further analytical work is being conducted utilizing a Light Scattering Detector to aid in the establishment of the correct molecular weight profile. Based on dextran standards, the leaf contains polysaccharides at 10-18 mg/g (dry) with no significant difference throughout the growth of the plant. The level in flower is slightly lower at 5-13 mg/g with indication of an increase during plant growth. It is of some interest that polysaccharides are present in flowers that have insenescence. These results are being re-examined and a correlation of the degree of senescence to the polysaccharides content being more closely observed. The root contains 3-10 mg/g during its first season of growth and a substantially higher amount (30-40 mg/g) at maturity after 18 months growth. These results reveal a substantial increase in yield may be obtained through an 18month growing period for the root section.

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Project Title

Commercial Borage Production for Oil and Gamma-linolenic Acid

RIRDC Project No:

UT-33A

Start Date: 01/09/00 Finish Date: 31/08/03 Researcher: Dr. Rowland Laurence Organisation: University of Tasmania

GPO Box 447 BURNIE TAS 7320


• The research proposed here will seek to improve agronomy and harvesting

of borage in order that the yields and profitability in this new Australian industry will allow grower and processor stakeholders to maximise their competitive advantage in an expanding world market.

• The key deliverable of the proposed work will be the provision to grower stakeholders of recommendations for the commercial production of borage in Tasmania/South East Australia. These will be synthesized from the verification and adaption of available overseas information through local field experimentation. The latter will concentrate on those husbandry practices, which are likely to be the major deterimants of yield and profitability.

Current Progress

This project, which began in September 2000, attempts to maximise commercially harvested yield, optimise planting time and find adequate methods of weed control.

Three field investigations have been carried out in 2001-02. A further small plot comparison of a range of planting times, from July 2001 through to October 2002, has been harvested and seed is presently being re-cleaned before analysis of the data to verify the reduction in yield with delayed sowing found in the previous season’s experiment.

While most emphasis is being placed on non-chemical methods for weed control, in recognition of market preferences, a range of ’best bet’ commercial herbicides has been applied, post emergence, to small plots of borage in the late rosette stage of growth. Of these herbicides, Allicide®, Stomp® and Ramrod® showed no visual effects on growth of borage.

In addition, a field trial has been carried out attempting to improve upon the minimal herbicide production regime investigated in the previous growing season. Replicated large plots, with a total area of 0.5 hectare, compared three sowing rates and two row spacings to assess crop competition with weeds after the use of a stale seed-bed technique, a pre-emergence contact herbicide and mechanical post emergence weed control with an inter-row brush weeder. The crop competed very well with weeds, which were confined to an insignificant understorey. Prolonged wet weather after windrowing of the borage in December prevented final commercial harvesting for two months. While seed re-cleaning is still to be completed, yields are likely to be higher than this delay in reclaiming the windrow earlier suggested.

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Project Title

Determination of pesticide minimum residue limits in essential oils

RIRDC Project No:

UT-36A




• To establish adequate withholding periods after pesticide application to

ensure sufficient dissipation of the active ingredients from essential oil crops

• To assess the efficiency of disposable SPE cartridges in the clean-up of essential oil.

• To develop analytical techniques using ECD and NPD • To produce an updated manual detailing more streamlined methodologies

for pesticide residue analyses in essential oils.

Current Progress

Peppermint and dill samples were collected several days after routine applications of pesticides. Despite the short time period between application and sample collection, no pesticide residues were detected. Propiconazole has previously been detected in peppermint crops at the final harvest. More comprehensive trials are planned for the 2002 season. The application of SPE to the clean-up of pesticide residues has been undertaken. Anion exchange discs were trialled for the separation of aqueous solutions of chelated mancozeb. The subsequent derivatisation with organic solvent based methylating reagents was successful but recoveries were poor compared to that obtained with phase transfer reagents. The discs were used to clean-up halogenated acidic pesticide residues. Detection was undertaken using GC ECD. Encouraging results were obtained but severe contamination of the GC column was evident. The source of contamination is being investigated. Analytical method development and validation is continuing. LCQ capabilities have been extended to include screening of boronia oil and blackcurrant concrete. Extensive work has been undertaken to detect residues of Dithane in boronia. In addition to the work on methylating chelated residues of mancozeb, vigorous acidic digestions to allow for the quantification of zinc and manganese using ICPMS has been undertaken with limited success. The oils produced in the 2002 season have been screened. Pesticide residues have been detected and samples are to be resubmitted for analyses to confirm results obtained.

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Project Title Production of a newsletter of the Essential Oil

Producers Association of Australia

RIRDC Project No:

PSE-2A

Start Date: 01/07/01 Finish Date: 30/06/03 Researcher: Dr. Erich Lassak Organisation: Phytochemical Services

254 Quarter Sessions Road WESTLEIGH NSW 2120

Phone: (02) 9875 1894 Fax: (02) 9875 1791 Objectives

• To bring together growers, producers, traders, researchers and users of

essential oils and of certain plant related natural products by the dissemination of information, which will stimulate the growth and development of Australian industries based on these products.

• It is proposed to produce three issues of the newsletter over two years.

Current Progress

The first newsletter for this project was published in March 2002. It included reports on the adoption of a tea tree oil monograph by the European Pharmacopoeia, reports on recent essential oils conferences and a listing of the botanical names of Australian native species used for commercial essential oil production. The next newsletter is due to be published in January 2003.

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Completed Projects Project Title:

Growing Milkweed, a plant with prospective anti-cancer properties

RIRDC Project No:

DAQ-281A

Researcher: Dr Craig Davis Organisation: Centre for Food Technology

19 Hercules Street, Hamilton 4007 Qld Phone: (07) 3406 8611 Fax: (07) 3406 8677 Email: [email protected] Objectives:

• To review the traditional use (and toxicity, where reported) of Euphorbia species in medicine and survey their potential for agricultural cultivation.

Background: Recent research by a small Brisbane-based, Australian-owned company Peplin Biotech

Ltd in conjunction with the Queensland Institute of Medical Research has found that extracts from milkweed (Euphorbia peplus) and potentially other Euphorbia species (e.g. E.esula, E.lathyris, E.terracina and E.lagascae) have significant anti-cancer activity in mice and humans. Euphorbia is a large genus of some 2000 species with milky sap, often with poisonous or medicinal uses. It includes beneficial as well as weedy plants, of which some 45 species are found in Australia. Although milkweed and the other promising Euphorbia spp. are new to agriculture, environmental requirements for their cultivation have been established in the course of earlier research by Peplin Biotech and QIMR. The active compounds are unable to be synthesised on a commercial scale and have been patented by Peplin Biotech. An early version of the product has given >90% complete responses on skin cancers in a Phase II clinical trial.

Research: A comprehensive literature review has been prepared with information being sourced from the resources of the Agricultural (DPI) and the Medical (QIMR) database systems as well as from the considerable library on the topic which is held by Peplin Biotech. This review discusses the presence of anti-cancer activity, the previous testing undertaken and the potential toxicity of the product.

Outcomes: Peplin Biotech has discovered a novel class of natural compounds, purified from Euphorbia, which show great potential in the laboratory as a potent treatment for a wide range of human cancers including breast, prostate cancer and skin cancers. An early clinical trial on thick and thin non-melanoma skin cancers has confirmed that the compounds are very effective in producing long-term (possibly permanent) responses in human patients without any evident systemic toxicity when applied topically. E.peplus is one of a number of plants in the Euphorbiaceae family that has attracted attention as a home remedy for skin cancer because of its milky sap. However, a survey by Peplin Biotech of over 200 species of the Euphorbiaceae family has shown that only E.peplus has the desired attributes of anti-cancer efficacy. In addition, E.peplus is not a noxious weed, it grows rapidly, produces harvestable seed and it is potentially suitable for large-scale agricultural production. Methods have been developed by Peplin Biotech for the extraction and purification of compounds from E.peplus in the laboratory, and these are currently being scaled-up for commercial production. It is anticipated that the overall cost of producing the active ingredient in pure form will result in a commercially viable treatment, provided the cost of agricultural production can be kept low. Currently, the plant is sown and harvested by hand and tended in small plots. The limiting factor in capturing commercial value from these discoveries is therefore the availability of plant feedstock for the extraction of active compounds. Developing large-scale agricultural production can solve this problem.

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Implications: Anti-cancer compounds from a common but tractable weed are being developed by a Brisbane-based, Australian-owned biotechnology company (Peplin Biotech) with the assistance of the Queensland Department of Primary Industries and the Queensland Institute of Medical Research. The present project may result in a unique opportunity for primary producers to contribute to their own wellbeing through the development of an effective treatment for non-melanoma skin cancer. A new, specialist crop might also emerge. We plan to evaluate methods for complete mechanisation of E.peplus cultivation and sap production. This will include methods of seed production, since the current supply of seed will need to be greatly expanded. The requirements for large-scale agriculture will be ascertained, using the expertise and equipment pooled from the wide variety of specialists in the DPI. The next critical step is to develop methods suitable for broad-acre, mechanised production.

16

Project Title:

Assessment of Chamomile Varieties for Essential Oil and Dried Flower Production

RIRDC Project No:

UT-28A

Researcher: Professor R. C. Menary Dr L. Falzari

Organisation: University of Tasmania Phone: 03 6226 2724 Fax: 03 6226 7609 Email: R. [email protected] Objectives

• The project objectives were the development of a production system for the

growing, harvesting and drying of high grade chamomile (Chamomilla recutita) flowers for the medicinal market and the distillation of the lower grade flowers for blue chamomile essential oil.

Background Chamomile is a new crop to be added to the armoury of the Essential Oils Industry in

Tasmania. Chamomile has been used in folk medicine throughout history. It is still used in the production of a medicinal tea that is renowned for its calming properties. The blue essential oil is used in the pharmaceutical and cosmetic industries. Production of chamomile in Tasmania will provide both import replacement and potentially a valuable export commodity.

Research The production of three varieties of chamomile was tested under Tasmanian growing conditions. The best sowing and harvest time was found for each variety. Yields of oil and flowers were measured and the best method for treating flowers before steam-distillation examined. The increase in oil yield from increasing the duration of steam-distillation was used to estimate the appropriate length of time of the distillation process.

Outcomes The optimum sowing time for chamomile is early spring. Multiple harvests optimise yield. The first harvest should coincide with the opening of the yellow disc-florets and the precise harvest date is determined by repeated sampling of the crop to measure increases in yield and quality. Yields of 1 tonne of dry flowers per hectare can be expected. Oil yield is strongly dependent on the duration of distillation and approximately 12 hours of distillation is recommended. Freezing is a suitable method of storing flowers before distillation or the flowers may be dried. Drying did not effect the quality of oil from the variety Bona but had a detrimental affect on the quality of steam-distilled oil from the variety Bodegold .

Implications The crop must be sown early and must be monitored closely to maximise yield and quality of both flowers and oil. No impediments were discovered which would preclude chamomile from being produced in Tasmania.

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Project Title:

Potential for IPM in Peppermint growing in South East Australia

RIRDC Project No:

DAV-178A

Researcher: Fred Bienvenu Organisation: NRE Ovens

Institute for Horticultural Development P.O. Box 235 Myrtleford 3737

Phone: 03-57 311 222 Fax: 03-57 311 223 Email: [email protected] Objectives

• To establish an interim protocol for minimising two-spotted mite (TSM) impact on south eastern Australia peppermint crops.

• To develop suitable sampling techniques for assessing TSM in peppermint crops.

• To evaluate the potential to develop an effective integrated pest management program for peppermint production in south eastern Australia based on effective control of TSM.

Background TSM is one of the most difficult horticultural pests to control and constitutes a very

significant and real risk to stable commercial peppermint oil production. The incidence of TSM is increasing annually in commercial peppermint crops. Significant yield losses that occurred in 1998/99 have been largely attributed to TSM. Under dry conditions high levels of TSM infestation result in excessive leaf loss, particularly lower leaf and can affect oil quality.

Research A field survey of commercial peppermint crops was carried out to establish which pests are present in peppermint fields and the range of controls currently used or available. Predators were released on to the two major peppermint properties and have proved adaptable to Victorian environmental conditions. Sampling and monitoring techniques were tested. Sampling for several predator generations was required to ensure that the life cycle would be complete.

Outcomes Evaluation of the potential to develop an effective integrated pest management program for peppermint production in south eastern Australia based on effective control of TSM has been successfully completed. The need for an IPM program for Australian peppermint is apparent and the many positive sightings indicate that the predator mite Phytoseiulus persimilis can survive and reduce TSM populations during the critical months of peppermint production.

Implications To implement a successful IPM program in peppermint several key developments are required:

• The identification and integration of chemical and sustainable bio-control methods practices that will maximise yield of high quality peppermint oil.

• The development of data required to support registration (minor use or full) of predator “friendly” pesticides

• Establishment of an industry reference group to facilitate and implement the use of a practical IPM program.

See progress report on the follow-up project DAV-190A (page 9).

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Project Title Methodology and viability of re-establishing commercial boronia plantations

RIRDC Project No:

SAG-1A

Researcher: Dr Doris Blaesing Organisation: Serve-Ag Pty Ltd Phone: (03) 6427 0800 Fax: (03) 6427 0801 Email: [email protected] Objectives

• This study will enable the boronia industry to determine the viability of replanting

present boronia plantations and indicate the management procedures necessary to successfully re-establish boronia plantations under their existing infrastructure.

Background The production of an essential oil from brown boronia (Boronia megastigma) has

been developed in Tasmania over the last 17 years, with commercial production of large clonal plantations for the last 12 years. At present, some 80% of the total Boronia has been in commercial production in Tasmania for more than 7 years, and plant numbers have been decreasing due to mechanical damage by harvesters and wind, and general decline. Production and sales forecasts from oil processors indicate that unless replanting is initiated in the next 1-3 years, there will be a shortfall in supplying the demand for product. Replanting boronia in small areas of plantations has not been successful to date, and growers are concerned about the viability of replanting entire plantations. The cost of developing new land rather than replanting current sites would be prohibitive. As boronia has very specific soil requirements, there are also no alternative cropping options for the land developed for current boronia production.

Research 1. Confirmation of specific replant disease (soil sickness) in a pot trial. 2. Isolation of soil-borne fungi from current boronia plantations. 3. Replant field trial, testing soil amendments, fire, nutrients simulating an ‘ash bed

effect’, and fungicides active against Phytophthora spp. 4. Field survey of Tasmanian boronia plantations to investigate site factors and plant

establishment, management and longevity. Outcomes • Classical ‘soil sickness’ problems have not been found.

• Several soil amendments and nutritional inputs can improve plant re-establishment.

• Sick plants may respond to targeted fungicide treatment. • A revision of the nutrition program may improve plant longevity. • Decline problems in current plantations are due to a combination of site specific,

establishment and management issues (water-logging, pot bound roots, irrigation, nutrient imbalance, scale and rust epidemics), and crown disease and deformation.

Implications • Treatments from the replant trial should be evaluated longer term at several sites.

• The importance of Phytophthora and Pythium as pathogens of cultivated boronia needs further investigation.

• Cation (Ca, K, Mg, NH4) and trace element nutrition as well as nutrient balances, need to be better understood to develop desirable plant nutrient levels that enable nutrient monitoring and adjustment of fertiliser programs for optimum production.

• Nematodes may play a role in plant decline that needs further investigation. • Factors affecting crown health, i.e. crown gall, have to be investigated and

managed. • Pesticides must not be used without thorough evaluation and use permits. • The industry may benefit from producing a comprehensive boronia management

guide and/or introducing a quality management system.

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Project Title:

Production of high quality Australian ginseng

RIRDC Project No:

UNC-8A

Researcher: Prof Ron Wills Organisation: University of Newcastle, Centre for Advancement of Food Technology & Nutrition

PO Box 127, OURIMBAH NSW 2258 Phone: (02) 4348 4140 Fax: (02) 4348 4145 Email: [email protected] Objectives

• To generate information on the active constituents in American ginseng that confer a health benefit to enable the Australian industry to position itself as an international supplier of high quality ginseng products.

Background American ginseng (Panax quinquefolium) is a medicinal herb native to North America but

is now widely used in traditional markets in Asia. The substantial international trade in ginseng led to the establishment of an Australian industry based on growing American ginseng. If Australia is to become successful in the exporting and import substitution of ginseng, it needs to develop a reputation for the consistent supply of high quality ginseng products. The ultimate determinant of quality is the concentration of active constituents that impart a health benefit to the human body. The group of saponins known as ginsenosides are widely accepted as major active constituents in ginseng.

Research The research focused on determining: reliable methods for the analysis of ginsenosides, changes in ginsenosides in plant parts during plant growth, effect of postharvest handling and processing operations on ginsenosides, quality of ginseng products in retail outlets, and a survey of practices of Australian ginseng farms.

Outcomes Quantitative analytical methods for the analysis of neutral and malonyl ginsenosides were developed using high performance liquid chromatography (HPLC). Ginseng roots were found to contain a similar composition and concentration of ginsenosides as plants grown in North America and Asia. The ginsenosides concentration in the root changed little over a growing season but was maximal in leaf and hair root when green fruit was present. Fully developed leaf contained 25% of total plant ginsenosides. Roots of different age or the same age but from different sites showed a linear relationship for root weight and ginsenosides concentration. Field cultivated plants under artificial shade produced faster growing roots and greater ginsenosides concentration than forest-grown roots. The drying of fresh roots in a hot air drier resulted in an increasing loss of total ginsenosides as the temperature was increased. Stored dried ginseng root was also temperature dependent with an increasing loss of product with a 2 hour blanch time and only a 15% loss in total ginsenosides. A wide range of ethanol/water mixtures extracted 85-90% of ginsenosides with 50% ethanol the optimal solvent. Spray drying the alcoholic extract gave a powder of acceptable colour and texture with <15% loss of ginsenosides. The quality of ginseng products available to consumers showed considerable variation in ginsenosides. The highest concentration was found in root powders and tea bags followed by the dry root and tablets/capsules.

Implications • The similar profile of ginsenosides in Australian-grown root crops to those grown in North America and Asia should support international marketing of the crop.

• Utilisation of the leaf could generate a valuable by-product with beneficial medicinal properties. Annual harvesting would allow a cash return before root maturity.

• Identification of growing practices or improved nutrition that increases the rate of root growth would appear to also give an increase in ginsenosides concentration.

• The temperature for drying and storing ginseng roots should be minimised to retain ginsenosides. Drying and storage temperatures of 55oC and 5oC are recommended.

• Production of a red ginseng product and a dried ginseng extract should be explored.

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Publications Presentations were made to the Annual Conference of the Australian Ginseng Growers Association held in 1999 at Traralgon, 2000 at Wangaratta and 2001 at Ballarat

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Project Title:

The Australian Lavender Industry - A Review of Oil Production and Related Products

RIRDC Project No:

SAG-2A

Researcher: Dr Lee Peterson Organisation: Serve-Ag Pty Ltd

PO Box 149 Glenorchy. TAS 7010


• To conduct a review of the Lavender Industry in Australia determining the

present status of the industry, current research projects and potential research and development issues.

Background Lavender oil is produced for a number of different markets. The traditional market

has been for fragrance applications, predominantly in body care products. The application of lavender oil in aromatherapy products has been steadily increasing despite the considerable historical use of lavender oil in aromatherapy. This increase appears to be driven by a renewed awareness in the potential therapeutic benefits of lavender oil

Research Much of the recent research in the rest of the world centres around the therapeutic activity of lavender based products. Most of these trials are targeted at determining the effect of lavender fragrance on the human emotional state. Other studies have examined the potential for lavender as a local anaesthetic. Another novel use for lavender oil that has been recently studied is the potential for the oil as a pesticide. Studies on the wound healing activities of L.angustifolia and L.allardii oils are being conducted by the Charles Sturt University (CSU). Within this research, lavender honey is also being examined as a wound dressing substrate. Other areas of Australian research focus on oil analysis and more in-depth examination of oils and their individual components. CSU has also been conducting headspace analysis of a range of varieties of lavender to look at the potential for chemotaxonomic differentiation of varieties whilst the University of Tasmania has examined the oil composition of the oil from Bridestowe Estate cultivars over a number of years. The University of Tasmania now holds a collection of very low camphor selections.

Outcomes The most significant area of research required for the Lavender industry is in regard to the breeding and selection aspects to further improve the genetic basis for lavender production. The high quality oil produced by Bridestowe Estate is a direct result of many decades of selection for this particular region of Tasmania. To enable successful oil production in other regions of Australia considerable emphasis will have to be placed on the selection and potential breeding of lavenders suitable for these regions to achieve quality standards and also economic yields. Further research and development of the therapeutic properties of lavender oil is a high priority for the industry. The majority of L.angustifolia oil market growth in the last 5-10 years has been in this segment rather than in the fragrance market

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Implications There is a growing demand for all of the lavender range of oil types – L. allardii and lavandin oils are sought after by the aromatherapy market, as is L.angustifolia oil, for their therapeutic applications. Currently oil produced for the therapeutic market must conform to the BP standard as products are mainly marketed into the aromatherapy segment. This presents difficulties for producers and distributors as not all oils have a BP standard eg: L. Allardii. The high incidence of incorrect identification and naming of species and varieties within the genus Lavandula further complicates the issue of standards. Many nurseries are incorrectly identifying various lavender species and labelling plants with only generic terms such as “English” or “French”. For prospective oil producers to source the correct cultivars, especially of L.angustifolia, this situation is totally unacceptable. In respect to the development of the industry associations, there is a definite requirement for an industry appointed representative or executive officer to be employed by the Lavender Growers Association (TALGA). Whilst the present volunteer committee is highly active and enthusiastic to further the development of the lavender industry in Australia, there exists a need for TALGA to be professionally managed and represented to industry and government bodies.

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