METHODS

IN

MOLECULAR BIOLOGY

Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hatfield, Hertfordshire, AL10 9AB, UK

For further volumes:

http://www.springer.com/series/7651

Natural Products Isolation

Third Edition

Edited by

Satyajit D. Sarker
Department of Pharmacy, School of Applied Sciences,
University of Wolverhampton, Wolverhampton, UK

Lutfun Nahar
Leicester School of Pharmacy, De Montfort University,
The Gateway, Leicester, UK

Editors
Satyajit D. Sarker, Ph.D.
Department of Pharmacy
School of Applied Sciences
University of Wolverhampton
Wolverhampton, UK

Lutfun Nahar, Ph.D.

Leicester School of Pharmacy
De Montfort University
Leicester, UK

ISSN 1064-3745
e-ISSN 1940-6029
ISBN 978-1-61779-623-4
e-ISBN 978-1-61779-624-1
DOI 10.1007/978-1-61779-624-1
Springer New York Dordrecht Heidelberg London
Library of Congress Control Number: 2012932311
Springer Science+Business Media, LLC 2012
All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the
publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA),
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Printed on acid-free paper
Humana Press is part of Springer Science+Business Media (www.springer.com)

Dedication
Dedicated to the memories of our dads.

Preface
The use of natural products for the treatment of various ailments can be traced back thousands
of years, and many block-buster drugs that we use today have come from natural origins.
Because of unique inherent chemical diversity, natural products have continued to be one
of the major sources of new drug molecules still today. In recent years, there has been a
remarkable resurgence of interests observed in the area of natural products research. With
significant developments in the areas of chromatography and spectroscopy, natural products research has gained new momentum. Natural products and their analogs are now suitable for high-throughput screening, and have become part of major and meaningful drug
discovery programs.
Isolation of natural products is one of the main components in natural products
research, especially in relation to drug discovery and development. Techniques for extraction and isolation of natural products from various matrices have evolved over the years,
and particularly, over the last couple of decades, the advances in extraction and isolation
technologies have made isolation of natural products less laborious and less time-consuming.
The last two editions of this book captured most of the techniques that are generally used
in natural products extraction and isolation. Since the publication of the second edition
in 2005, there has been significant progress in these areas, which certainly demands the
publication of this third edition to document new methods and technologies for natural
products isolation.
This thoroughly revised third edition has incorporated a total of 20 chapters; some are
new chapters, while some others are expanded or updated version of the chapters from the
second edition, using a slightly new lay-out and style. Chapters in this edition have included
a number of hands on approaches and step-by-step protocols to help natural products
researchers follow and implement these protocols in their own research. Like the previous
editions, it is still our intention to provide substantial background information for budding
natural product researchers as well as to offer reference guide to available methodologies
and techniques for the more experienced researchers.
Wolverhampton, UK
Leicester, UK

Satyajit D. Sarker
Lutfun Nahar

vii

Contents
Dedication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

v
vii
xi

1 An Introduction to Natural Products Isolation . . . . . . . . . . . . . . . . . . . . . . . .

Satyajit D. Sarker and Lutfun Nahar
2 Initial and Bulk Extraction of Natural Products Isolation. . . . . . . . . . . . . . . . .
Vronique Seidel
3 Supercritical Fluid Extraction in Natural Products Analyses . . . . . . . . . . . . . . .
Lutfun Nahar and Satyajit D. Sarker
4 Accelerated Solvent Extraction for Natural Products Isolation . . . . . . . . . . . . .
Mohammad A. Mottaleb and Satyajit D. Sarker
5 Microwave-Assisted Extraction in Natural Products Isolation. . . . . . . . . . . . . .
Abbas Delazar, Lutfun Nahar, Sanaz Hamedeyazdan,
and Satyajit D. Sarker
6 An Introduction to Planar Chromatography and Its Application
to Natural Products Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Simon Gibbons
7 Isolation of Natural Products by Low-Pressure
Column Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Raymond G. Reid and Satyajit D. Sarker
8 Isolation of Natural Products by Ion-Exchange Methods. . . . . . . . . . . . . . . . .
Klaus Dragull and John J. Beck
9 Separation of Natural Products by Countercurrent Chromatography . . . . . . . .
James B. McAlpine, J. Brent Friesen, and Guido F. Pauli
10 Isolation of Natural Products by Preparative High Performance
Liquid Chromatography (Prep-HPLC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Zahid Latif and Satyajit D. Sarker
11 Isolation of Natural Products by Preparative Gas Chromatography . . . . . . . . .
Temel zek and Fatih Demirci
12 Hyphenated Techniques and Their Applications in Natural
Products Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Satyajit D. Sarker and Lutfun Nahar
13 Extraction of Plant Secondary Metabolites . . . . . . . . . . . . . . . . . . . . . . . . . . .
William P. Jones and A. Douglas Kinghorn
14 Isolation of Marine Natural Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Wael E. Houssen and Marcel Jaspars

ix

27
43
75
89

117

155
189
221

255
275

301
341
367

Contents

15 Isolation of Microbial Natural Products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Olov Sterner
16 Extraction and Isolation of Saponins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Runner R.T. Majinda
17 Extraction and Isolation of Phenolic Compounds . . . . . . . . . . . . . . . . . . . . . .
Celestino Santos-Buelga, Susana Gonzalez-Manzano,
Montserrat Dueas, and Ana M. Gonzalez-Paramas
18 Scaling-Up of Natural Products Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Yash Kumarasamy
19 Follow-Up of Natural Products Isolation. . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Richard J.P. Cannell, Satyajit D. Sarker, and Lutfun Nahar
20 Natural Products Isolation in Modern Drug Discovery Programs . . . . . . . . . .
Alexander I. Gray, John O. Igoli, and RuAngelie Edrada-Ebel
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

393
415
427

465
473
515
535

Contributors
JOHN J. BECK Plant Mycotoxin Research, USDA-ARS, WRRC, Albany, CA, USA
RICHARD J.P. CANNELL (LATE)
ABBAS DELAZAR School of Pharmacy and Drug Applied Research Center,
Tabriz University of Medical Sciences, Tabriz, Iran
FATIH DEMIRCI Department of Pharmacognosy, Anadolu University, Eskisehir, Turkey
KLAUS DRAGULL Plant Mycotoxin Research, USDA-ARS, WRRC, Albany, CA, USA
MONTSERRAT DUEAS Grupo de Investigacin de Polifenoles (GIP-USAL),
Facultad de Farmacia, Universidad de Salamanca, Salamanca, Spain
RUANGELIE EDRADA-EBEL Natural Products Research Laboratories,
Strathclyde Institute of Pharmacy and Biomedical Sciences,
University of Strathclyde, Glasgow, UK
J. BRENT FRIESEN Department of Medicinal Chemistry and Pharmacognosy,
School of Pharmacy, University of Illinois at Chicago, Chicago, IL, USA
SIMON GIBBONS Department of Pharmaceutical and Biological Chemistry,
The School of Pharmacy, UCL School of Pharmacy, London, UK
SUSANA GONZALEZ-MANZANO Grupo de Investigacin de Polifenoles (GIP-USAL), Facultad de Farmacia, Universidad de Salamanca, Salamanca, Spain
ANA M. GONZALEZ-PARAMAS Grupo de Investigacin de Polifenoles (GIP-USAL), Facultad de Farmacia, Universidad de Salamanca, Salamanca, Spain
ALEXANDER I. GRAY Natural Products Research Laboratories, Strathclyde Institute
of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK
SANAZ HAMEDEYAZDAN School of Pharmacy and Drug Applied Research Center, Tabriz
University of Medical Sciences, Tabriz, Iran
WAEL E. HOUSSEN Department of Chemistry, Marine Biodiscovery Centre,
University of Aberdeen, Old Aberdeen, UK
JOHN O. IGOLI Natural Products Research Group, BCDD, Strathclyde Institute
of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK
MARCEL JASPARS Department of Chemistry, Marine Biodiscovery Centre,
University of Aberdeen, Old Aberdeen, UK
WILLIAM P. JONES Ingredient and Process Research, Kraft Foods, Glenview, IL, USA
A. DOUGLAS KINGHORN Division of Medicinal Chemistry and Pharmacognosy,
College of Pharmacy, The Ohio State University, Columbus, OH, USA
YASH KUMARASAMY School of Pharmacy, The Robert Gordon University, Aberdeen, UK
ZAHID LATIF Technology Strategy Board, Swindon, UK
RUNNER R.T. MAJINDA Department of Chemistry, University of Botswana,
Gaborone, Botswana
JAMES B. MCALPINE Department of Medicinal Chemistry and Pharmacognosy,
School of Pharmacy, University of Illinois at Chicago, Chicago, IL, USA

xi

xii

Contributors

MOHAMMAD A. MOTTALEB Department of Chemistry/Physics, Center for Innovation and

Entrepreneurship, Northwest Missouri State University, Maryville, MO, USA
LUTFUN NAHAR Leicester School of Pharmacy, De Montfort University,
The Gateway, Leicester, UK
TEMEL ZEK Department of Pharmacognosy, Anadolu University, Eskisehir, Turkey
GUIDO F. PAULI Department of Medicinal Chemistry and Pharmacognosy,
School of Pharmacy, University of Illinois at Chicago, Chicago, IL, USA
RAYMOND G. REID School of Pharmacy, The Robert Gordon University, Aberdeen, UK
CELESTINO SANTOS-BUELGA Grupo de Investigacin de Polifenoles (GIP-USAL),
Facultad de Farmacia, Universidad de Salamanca, Salamanca, Spain
SATYAJIT D. SARKER Department of Pharmacy, School of Applied Sciences,
University of Wolverhampton, Wolverhampton WV11LY, UK
VRONIQUE SEIDEL Natural Products Research Laboratories, Strathclyde Institute
of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK
OLOV STERNER Department of Chemistry, Lund University, Lund, Sweden