BASIL Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. Medicinal and Aromatic Plants—Industrial Profiles Individual volumes in this series provide both industry and academia with in-depth coverage of one major medicinal or aromatic plant of industrial importance. Edited by Dr Roland Hardman Volume 1 Valerian edited by Peter J.Houghton Volume 2 Perilla edited by He-Ci Yu, Kenichi Kosuna and Megumi Haga Volume 3 Poppy edited by Jeno Bernáth Volume 4 Cannabis edited by David T.Brown Volume 5 Neem H.S.Puri Volume 6 Ergot edited by Vladimír K řen and Ladislav Cvak Volume 7 Caraway edited by Éva Németh Volume 8 Saffron edited by Moshe Negbi Volume 9 Tea Tree edited by lan Southwell and Robert Lowe Volume 10 Basil edited by Raimo Hiltunen and Yvonne Holm Other volumes in preparation Allium, edited by K.Chan Artemisia, edited by C.Wright Cardamom, edited by P.N.Ravindran and K.J.Madusoodanan Chamomile, edited by R.Franke and H.Schilcher Please see the back of this book for other volumes in preparation in Medicinal and Aromatic Plants—Industrial Profiles Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. BASIL The Genus Ocimum Edited by Raimo Hiltunen and Yvonne Holm Department of Pharmacy, University of Helsinki, Finland harwood academic publishers Australia • Canada • China • France • Germany • India • Japan Luxembourg • Malaysia • The Netherlands • Russia • Singapore Switzerland Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. This edition published in the Taylor & Francis e-Library, 2006. “To purchase your own copy of this or any of Taylor & Francis or Routledge’s collection of thousands of eBooks please go to www.eBookstore.tandf.co.uk.” Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. All rights reserved. No part of this book may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and recording, or by any information storage or retrieval system, without permission in writing from the publisher. Printed in Singapore. Amsteldijk 166 1st Floor 1079 LH Amsterdam The Netherlands British Library Cataloguing in Publication Data A catalogue record for this book is available from the British Library. ISBN 0-203-30377-6 Master e-book ISBN ISBN 0-203-34321-2 (Adobe eReader Format) ISBN 90-5702-432-2 (Print Edition) ISSN 1027–4502 Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. CONTENTS Preface to the Series vii Preface ix Contributors xi 1 Ocimum: An Overview of Classification and Relationships 1 Alan Paton R.M.Harley M.M.Harley 2 Production Systems of Sweet Basil 39 Eli Putievsky Bertalan Galambosi 3 Chemical Composition of Ocimum Species 67 Raimo Hiltunen 4 Essential Oil of Ocimum 77 Raimo Hiltunen Yvonne Holm 5 Bioactivity of Basil 113 Yvonne Holm 6 Processing and Use of Basil in Foodstuffs, Beverages and in Food Preparation 137 Seija Marjatta Mäkinen Kirsti Kaarina Pääkkönen Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. PREFACE TO THE SERIES There is increasing interest in industry, academia and the health sciences in medicinal and aromatic plants. In passing from plant production to the eventual product used by the public, many sciences are involved. This series brings together information which is currently scattered through an ever increasing number of journals. Each volume gives an in-depth look at one plant genus, about which an area specialist has assembled information ranging from the production of the plant to market trends and quality control. Many industries are involved such as forestry, agriculture, chemical, food, flavour, beverage, pharmaceutical, cosmetic and fragrance. The plant raw materials are roots, rhizomes, bulbs, leaves, stems, barks, wood, flowers, fruits and seeds. These yield gums, resins, essential (volatile) oils, fixed oils, waxes, juices, extracts and spices for medicinal and aromatic purposes. All these commodities are traded world-wide. A dealer’s market report for an item may say “Drought in the country of origin has forced up prices”. Natural products do not mean safe products and account of this has to be taken by the above industries, which are subject to regulation. For example, a number of plants which are approved for use in medicine must not be used in cosmetic products. The assessment of safe to use starts with the harvested plant material which has to comply with an official monograph. This may require absence of, or prescribed limits of, radioactive material, heavy metals, aflatoxins, pesticide residue, as well as the required level of active principle. This analytical control is costly and tends to exclude small batches of plant material. Large scale contracted mechanised cultivation with designated seed or plantlets is now preferable. Today, plant selection is not only for the yield of active principle, but for the plant’s ability to overcome disease, climatic stress and the hazards caused by mankind. Such methods as in vitro fertilisation, meristem cultures and somatic embryogenesis are used. The transfer of sections of DNA is giving rise to controversy in the case of some enduses of the plant material. Some suppliers of plant raw material are now able to certify that they are supplying organically-farmed medicinal plants, herbs and spices. The Economic Union directive (CVO/EU No 2092/91) details the specifications for the obligatory quality controls to be carried out at all stages of production and processing of organic products. Fascinating plant folklore and ethnopharmacology leads to medicinal potential. Examples are the muscle relaxants based on the arrow poison, curare, from species of Chondrodendron, and the antimalarials derived from species of Cinchona and Artemisia. The methods of detection of pharmacological activity have become increasingly reliable and specific, frequently involving enzymes in bioassays and avoiding the use of laboratory animals. By using bioassay linked fractionation of crude plant juices or extracts, compounds can be specifically targeted which, for example, inhibit blood platelet aggregation, or have antitumour, or antiviral, or any other required activity. With the assistance of robotic devices, all the members of a genus may be readily screened. However, the plant material must be fully authenticated by a specialist. vii Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. viii PREFACE TO THE SERIES The medicinal traditions of ancient civilisations such as those of China and India have a large armamentarium of plants in their pharmacopoeias which are used throughout South East Asia. A similar situation exists in Africa and South America. Thus, a very high percentage of the world’s population relies on medicinal and aromatic plants for their medicine. Western medicine is also responding. Already in Germany all medical practitioners have to pass an examination in phytotherapy before being allowed to practise. It is noticeable that throughout Europe and the USA, medical, pharmacy and health related schools are increasingly offering training in phytotherapy. Multinational pharmaceutical companies have become less enamoured of the single compound magic bullet cure. The high costs of such ventures and the endless competition from me too compounds from rival companies often discourage the attempt. Independent phytomedicine companies have been very strong in Germany. However, by the end of 1995, eleven (almost all) had been acquired by the multinational pharmaceutical firms, acknowledging the lay public’s growing demand for phytomedicines in the Western World. The business of dietary supplements in the Western World has expanded from the Health Store to the pharmacy. Alternative medicine includes plant based products. Appropriate measures to ensure the quality, safety and efficacy of these either already exist or are being answered by greater legislative control by such bodies as the Food and Drug Administration of the USA and the recently created European Agency for the Evaluation of Medicinal Products, based in London. In the USA, the Dietary Supplement and Health Education Act of 1994 recognised the class of phytotherapeutic agents derived from medicinal and aromatic plants. Furthermore, under public pressure, the US Congress set up an Office of Alternative Medicine and this office in 1994 assisted the filing of several Investigational New Drug (IND) applications, required for clinical trials of some Chinese herbal preparations. The significance of these applications was that each Chinese preparation involved several plants and yet was handled as a single IND. A demonstration of the contribution to efficacy, of each ingredient of each plant, was not required. This was a major step forward towards more sensible regulations in regard to phytomedicines. My thanks are due to the staff of Harwood Academic Publishers who have made this series possible and especially to the volume editors and their chapter contributors for the authoritative information. Roland Hardman Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. PREFACE Our interest in the plant named basil began in the early 1980s when the Division of Pharmacognosy at the University of Helsinki was involved in a cultivation project with aromatic plants. Basil is a tender herb, not very suitable for the Finnish climate. It could be successfully cultivated in greenhouse conditions though. Some years later Mr Galambosi brought seeds of different cultivars of Ocimum basilicum, cultivated the plants and made us analyse their essential oils. After that we were stuck in the fascinating world of basil oil. When searching the literature for data on the chemical composition one could not help noticing the confusion in the nomenclature. Thus a need to clarify the taxonomy and nomenclature was born, and through Mr Galambosi we got into contact with Dr Paton, who accepted the task and now provides a delimitation of Ocimum from related genera and an infrageneric classification of Ocimum. Chemotaxonomy has been used as a tool to separate the different species. However, it has not been very successful because there are a large number of subspecies, varieties, forms, cultivars and even some hybrids. These are not readily separated on the basis of the essential oil composition. In addition, the essential oil composition is known to vary depending on the cultivation methods, drying and isolation techniques used. The establishment of chemotypes within the species is not easily performed either. There is always one oil which will not fit into an already established group. Ocimum sanctum or Holy Basil is a sacred medicinal plant in India, where it is used for many different ailments in ethnomedicine. Some activities have been confirmed by pharmacological studies. O.gratissimum, O.viride and O.suave are native plants in different parts of Africa and are used in traditional medicine, mostly as expectorants. The essential oils of these species also exhibit large antimicrobial spectra. An important activity is the insect repelling effect of many Ocimum oils, which can be utilized in warm countries. The composition of the fixed oil of Ocimum seeds has been studied lately and it was found to have anti-inflammatory activity. In spite of all the pharmacological activities of Ocimum essential oils the main use of the plants is as aromatic plants and spices. This book is intended to cover the present knowledge of all aspects of the cultivation, composition and use of Ocimum plants. There certainly are areas which need further exploitation, such as the enantiomeric composition of the essential oils. We hope this book will fulfil your expectations regarding its contents. Raimo Hiltunen Yvonne Holm ix Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. CONTRIBUTORS Bertalan Galambosi Seija Marjatta Mäkinen Agricultural Research Centre of Finland Department of Applied Chemistry Karila Research Station for Ecological and Microbiology Agriculture Division of Nutrition Karilantie 2 A P.O. Box 27 FIN 50600 Mikkeli University of Helsinki Finland FIN 00014 Helsinki Finland M.M.Harley Herbarium Royal Botanic Gardens Kew Kirsti Kaarina Pääkkönen Richmond Department of Food Technology Surrey TW9 3AB P.O. Box 27 UK University of Helsinki FIN 00014 Helsinki R.M.Harley Finland Herbarium Royal Botanic Gardens Kew Richmond Alan Paton Surrey TW9 3AB Herbarium UK Royal Botanic Gardens Kew Raimo Hiltunen Richmond Department of Pharmacy Surrey TW9 3AB P.O.Box 56 UK University of Helsinki FIN 00014 Helsinki Finland Yvonne Holm Eli Putievsky Department of Pharmacy Agricultural Research Organization P.O. Box 56 Newe Ya’ar Research Centre University of Helsinki P.O. Box 90000 FIN 00014 Helsinki Haifa 31900 Finland Israel xi Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group. 1. OCIMUM: AN OVERVIEW OF CLASSIFICATION AND RELATIONSHIPS ALAN PATON, M.R.HARLEY and M.M.HARLEY Herbarium, Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3AB, UK SUMMARY The taxonomy and nomenclature of Ocimum are in a state of confusion. The aim of this paper is to clearly delimit Ocimum from related genera, provide an up- to-date infrageneric classification which can be used as a framework for understanding relationships within the genus, provide a list of recognised species with their correct names and common synonyms and a key to their identification. In all 64 species are recognised. A parsimony analysis was carried out which suggests Ocimum is a monophyletic group if segregate genera such as Becium and Erythrochlamys are included within it. An account of the morphological features of Ocimum is provided and the economic uses of the genus are briefly discussed in the context of the infrageneric classification. INTRODUCTION Ocimum L. is a member of the Labiatae family. The typical characteristics of this family are a square stem, opposite and decussate leaves with many gland dots. The flowers are strongly zygomorphic with two distinct lips. Many of the family, particularly subfamily Nepetoideae, to which Ocimum belongs, are strongly aromatic due to essential oils which consist of monoterpenes, sesquiterpenes and phenylpropanoids. Ocimum, unlike other economically important herbs in the Labiatae such as Rosmarinus, Thymus and Salvia, belongs in tribe Ocimeae which has declinate stamens. That is the stamens lie over the lower (anterior) lip of the corolla rather than ascending under the upper (posterior) lip. The Ocimeae are essentially a tropical tribe and Ocimum occurs naturally in tropical America, Africa and Asia. Unlike several other economic Labiatae, Ocimum requires warmth for growth and should be protected from frost. Ocimum is an important economic and medicinal herb, and yet its taxonomy and nomenclature are in a bit of a muddle. Taxonomy underpins all plant science: we must know the correct name for a plant if we are to communicate information about its uses and relationships. It is important that each species has only one correct name to avoid confusion and aid clarity. The International Code of Botanical Nomenclature (Greuter et al., 1994) ensures that names are attached by a standard set of rules. In the literature concerning Ocimum, particularly the industrial and economic papers, these rules have 1 Copyright © 1999 OPA (Overseas Publishers Association) N.V.Published by license under the Harwood Academic Publishers imprint, part of the Gordon and Breach Publishing Group.