Stevia

© 2002 Taylor & Francis

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 Jenö Bernáth
Volume 4
Cannabis, edited by David T.Brown
Volume 5
Neem, edited by H.S.Puri
Volume 6
Ergot, edited by Vladimír Kren and Ladislav Cvak
Volume 7
Caraway, edited by Éva Németh
Volume 8
Saffron, edited by Moshe Negbi
Volume 9
Tea Tree, edited by Ian Southwell and Robert Lowe
Volume 10
Basil, edited by Raimo Hiltunen and Yvonne Holm
Volume 11
Fenugreek, edited by Georgios Petropoulos
Volume 12
Gingko biloba, edited by Teris A.Van Beek
Volume 13
Black Pepper, edited by P.N.Ravindran
Volume 14
Sage, edited by Spiridon E.Kintzios
Volume 15
Ginseng, edited by W.E.Court
Volume 16
Mistletoe, edited by Arndt Büssing
Volume 17
Tea, edited by Yong-su Zhen
Volume 18
Artemisia, edited by Colin W.Wright
Volume 19
Stevia, edited by A.Douglas Kinghorn

© 2002 Taylor & Francis

Stevia
The genus Stevia

Edited by
A.Douglas Kinghorn
Department of Medicinal Chemistry and Pharmacognosy
University of Illinois at Chicago
USA

London and New York

© 2002 Taylor & Francis

First published 2002
by Taylor & Francis
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Simultaneously published in the USA and Canada
by Taylor & Francis Inc,
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© 2002 Taylor & Francis
All rights reserved. No part of this book may be reprinted or reproduced or utilised in
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authors can accept any legal responsibility or liability for any errors or omissions that
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© 2002 Taylor & Francis

Contents

Preface to the series vii

Preface ix
Acknowledgements xi
List of contributors xii

1 Overview 1
A.DOUGLAS KINGHORN

2 Botany of Stevia and Stevia rebaudiana 18

DJAJA DJENDOEL SOEJARTO

3 Ethnobotany of Stevia and Stevia rebaudiana 40

DJAJA DJENDOEL SOEJARTO

4 Sweet and non-sweet constituents of Stevia rebaudiana 68

EDWARD J.KENNELLY

5 The phytochemistry of Stevia: a general survey 86

CARLOS M.CERDA-GARCÍA-ROJAS AND ROGELIO PEREDA-MIRANDA

6 Synthetic investigations on steviol, stevioside, and

rebaudioside A, and their applications as starting materials 119
DARRICK S.H.L.KIM

7 Methods to improve the taste of the sweet principles of Stevia rebaudiana 138
KAZUHIRO OHTANI AND KAZUO YAMASAKI

8 Pharmacology and toxicology of stevioside, rebaudioside A, and steviol 160

RYAN J.HUXTABLE

9 Use of Stevia rebaudiana sweeteners in Japan 178

KENJI MIZUTANI AND OSAMU TANAKA

10 Use of stevioside and cultivation of Stevia rebaudiana in Korea 196

JINWOONG KIM,YOUNG HAE CHOI AND YOUNG-HEE CHOI

© 2002 Taylor & Francis

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 worldwide. 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, aflatoxin, 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 fertilization, meristem cultures and somatic embryogenesis are used. The transfer of
sections of DNA is giving rise to controversy in the case of some end-uses 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 anti-malarials 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 anti-tumour, or anti-viral, 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 fully authenticated by a specialist.

© 2002 Taylor & Francis

viii Preface to the series

The medicinal traditions of ancient civilisations such as those of China and India have a
large armamentaria 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
phyto-medicine 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 staffs of Harwood Academic Publishers and Taylor & Francis
who have made this series possible and especially to the volume editors and their chapter
contributors for the authoritative information.
Roland Hardman

© 2002 Taylor & Francis

Preface

The South American species Stevia rebaudiana (Bertoni) Bertoni (Compositae) is an

economically important plant which has attracted considerable controversy. In accumulating
in its leaves very high concentration levels of the sweet-tasting secondary metabolites stevioside
and rebaudioside A, S. rebaudiana is something of a chemotaxonomic curiosity. What makes
this species controversial is that products made from the refined leaf extracts or the pure
diterpene glycoside stevioside are used to substitute for sucrose in certain countries such as
Japan and Korea, but have been subjected to past or present restrictive governmental legislation
in other countries such as the United Kingdom and the United States. In the last century,
well over 1,000 scientific articles and patents on S. rebaudiana and its sweet glycosidic constituents
have been published, with some of these in languages other than English.While many technical
aspects have been covered in this large body of literature, the three most frequently covered
topics have been purification techniques for the S. rebaudiana sweet constituents, procedures
for improving the quality of the sweet-taste response elicited by stevioside, and in vitro and in
vivo safety studies on these natural sweeteners.
In the initial overview chapter of this volume, a number of aspects not dealt with later are
presented, such as the history of the major scientific and regulatory developments which have
led to the use of stevioside and its diterpene glycoside analogues as sweetening and flavoring
agents. Stevioside is compared to other ‘high intensity’ natural sweetness in terms of its sweetness
potency and other parameters.The physical properties of the sweet S. rebaudiana ent-kaurene
glycosides are considered, as well as a summary of the various types of analytical methods
developed for these compounds. Also discussed are the biosynthesis of steviol and stevioside,
with information also provided on the cultivation, cell culture, and commercial production of
S. rebaudiana. Finally, the potential of stevioside and its congeners in relation to dental caries
is reviewed, and a summary of the worldwide regulatory status of stevioside as a sweetener is
provided.
Chapters 2 and 3 deal in turn with the botany and ethnobotany of the genus Stevia as a
whole and S. rebaudiana in particular.The first of these chapters describes the common botanical
traits of the genus Stevia, which consists of 220–230 species and is entirely confined to North
and South America. The ethnobotanical treatment traces back the relationship of the genus
Stevia to man from the sixteenth century onwards, with particular emphasis on the early use
of S. rebaudiana for sweetening by indigenous Paraguayan populations.
In the first of four chapters on phytochemical, chemical and biochemical aspects of the S.
rebaudiana sweeteners, Chapter 4 provides a full listing of the presently known constituents of
this species, inclusive of triterpenoids and steroids, flavonoids, and miscellaneous compounds,
in addition to the diterpenoids. Chapter 5 summarizes the extensive phytochemical literature
on Stevia species other than S. rebaudiana, with particular emphasis on the wide range of
functionalized sesquiterpenoids that are elaborated by the members of this genus. In Chapter
6, procedures for the chemical synthesis of steviol and for the glycosylation of steviol to form

© 2002 Taylor & Francis

x Preface

stevioside are reviewed. In addition, synthetic applications using stevioside and steviol as
starting materials are presented. In Chapter 7, chemical and biochemical methods developed
in Japan to improve the taste of stevioside and the other sweet principles of S. rebaudiana are
described.
Chapter 8 provides an in-depth evaluation of the safety of extracts of S. rebaudiana as well
as the pure compounds stevioside and rebaudioside A, and their common ent-kaurene aglycone,
steviol. Finally, in Chapters 9 and 10, the use of refined S. rebaudiana extracts and stevioside in
Japan and Korea, respectively, is presented. The longest and most widespread use of products
from S. rebaudiana have been in Japan, for the sweetening and flavoring of a many different
types of food products. However, more recently, pure stevioside has been used increasingly as
a sucrose substitute in Korea, particularly for the sweetening of a traditional distilled liquor
called soju.
It is hoped that readers of this edited volume on S. rebaudiana and its sweet principles will
be able to gain a better understanding of a fascinating topic from a wide perspective as a result
of the various chapters provided, which have been written by acknowledged experts in each
area. It seems likely that the current widespread interest in stevioside, rebaudioside A, and
their various modified structural forms, will continue well into the twenty-first century.
A.Douglas Kinghorn

© 2002 Taylor & Francis

Acknowledgements

The Editor is grateful to all of the chapter authors, for lending their time and expertise to this
project. In addition, he thanks the Book Series Editor, Dr Roland Hardman for his unwavering
encouragement. Dr Aiko Ito is thanked for her helpful assistance in the editing of two of the
chapters and the compilation of the index. Finally, the following are thanked for providing
valuable information: Professors Jan M.C.Guens, Pier-Giorgio Pietta, Finn Sand-berg and
Vincente Oliverira Ferro, and Drs Gloria L.Silva and Luisella Verotta.

© 2002 Taylor & Francis

List of contributors

Carlos M.Cerda-García-Rojas Kwanak-ku

Departamento de Química Seoul 151–742, Korea
CINVESTAV
Institute Politécnico Nacional A.Douglas Kinghorn
Apartado Postal 14–740 Department of Medicinal Chemistry and
México, D.F. 07000, Mexico Pharmacognosy
College of Pharmacy (M/C 781)
Young Hae Choi University of Illinois at Chicago
College of Pharmacy 833 South Wood Street
Seoul National University Chicago, IL 60612, USA
San 56–1, Sinlim-dong
Kwanak-ku Rogelio Pereda-Miranda
Seoul 151–742, Korea Departamento de Farmacia
Facultad de Quimica
Young-Hee Choi Universidad Nacional Autónoma de México
College of Pharmacy Coyaocan México D.F., 04510 Mexico
Ewha Women’s University
11–1 Daehyun-dong, Seodaemun-gu Kenji Mizutani
Seoul 120–750, Korea Research and Development Division
Maruzen Pharmaceuticals Co., Ltd.
Ryan J.Huxtable 14703–10 Mukaihigashi-cho
Department of Pharmacology Onomichi, Hiroshima 722–0062, Japan
College of Medicine
University of Arizona Kazuhiro Ohtani
P.O. Box 245050 Institute of Pharmaceutical Sciences
Tucson, AZ 85724–5050 School of Medicine
Hiroshima University
Edward J.Kennelly 1–2–3 Kasumi, Minami-ku
Formerly Emeritus Professor Hiroshima 734–8551, Japan
U.S. Food and Drug Administration
Center for Food Safety and Applied Djaja Djendoel Soejarto
Nutrition Program for Collaborative Research in the
Office of Special Nutritionals Pharmaceutical Sciences
HFS-465, 200 C Street S.W. College of Pharmacy (M/C 877)
Washington, DC 20204, USA University of Illinois at Chicago
Present address: 833 South Wood Street
Department of Biological Sciences Chicago, IL 60612, USA
Lehman College and
City University of New York Botany Department, Field Museum
250 Bedford Park Boulevard West Roosevelt Road at Lake Shore Drive
Bronx, NY 10468, USA Chicago, IL 60605, USA

Darrick S.H.L.Kim Osamu Tanaka

Program for Collaborative Research in the Emeritus Professor
Pharmaceutical Sciences and Department Institute of Pharmaceutical Sciences
of Medicinal Chemistry and School of Medicine
Pharmacognosy Hiroshima University
College of Pharmacy (M/C 877) 1–2–3 Kasumi, Minami-ku
University of Illinois at Chicago Hiroshima 734–8551, Japan
833 South Wood Street
Chicago, IL 60612, USA Kazuo Yamasaki
Institute of Pharmaceutical Sciences
Jinwoong Kim School of Medicine
College of Pharmacy Hiroshima University
Seoul National University 1–2–3 Kasumi, Minami-ku
San 56–1, Sinlim-dong Hiroshima 734–8551, Japan

© 2002 Taylor & Francis