Saponins Types Sources and Research

BIOCHEMISTRY RESEARCH TRENDS
SAPONINS
TYPES, SOURCES AND RESEARCH
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SAPONINS
TYPES, SOURCES AND RESEARCH
CANDICE GREENE
EDITOR
New York
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Library of Congress Cataloging-in-Publication Data

Names: Greene, Candice, editor.
Title: Saponins : types, sources and research / editor, Candice Greene.
Description: Hauppauge, New York : Nova Science Publisher's, Inc., [2016] |
Series: Biochemistry research trends | Includes bibliographical references
and index.
Identifiers: LCCN 2016044255 (print) | LCCN 2016045869 (ebook) | ISBN
9781536102895 (hardcover) | ISBN 9781536103113 (ebook) | ISBN
9781536103113
Subjects: LCSH: Saponins.
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Published by Nova Science Publishers, Inc. † New York

CONTENTS
Preface vii
Chapter 1 Saponins: Occurrence in Nature and
Biological Activities 1
Andresa Heemann Betti, Juliane Deise Fleck
and Simone Gasparin Verza
Chapter 2 Mini-Review: Ginsenoside Rg3 Modulating
Lipid Metabolism 37
Yong-Seob Jeong and Jisun Oh
Chapter 3 Application of Saponins in Remediation
of Metal-Contaminated Soils 63
Zygmunt Mariusz Gusiatin
Chapter 4 Application of Saponin from Soapnuts
for Remediation of Oil Contaminated Sand 91
Meshari S. Almutairi
Chapter 5 Triterpenoids and Their Saponins from Ilex asprella:
Types and Biological Studies 107
Hui Liu, Fan-Cheng Meng, Ying Wang,
Rui-Bing Wang, Wen-Cai Ye and Qing-Wen Zhang
Index 131
PREFACE
Saponins are surfactive compounds that are widely distributed in nature,

occurring primarily in the plant kingdom. Plants that produce saponins
generally accumulate these metabolites as part of their development, which
can be influenced by several environmental factors, such as nutrient and water
availability, light irradiation and/or a combination of these factors. In this
book, Chapter One discusses the occurrence in nature and biological activities
of saponins. Chapter Two provides a review of ginsenoside Rg3 modulating
lipid metabolism. Chapter Three demonstrates research on saponin application
in remediation of metal-contaminated soils, and methods of saponins recovery
after soil washing and attempts of their reuse. Chapter Four evaluates the
feasibility of using saponin from soapnuts (Sapindus mukorrossi) for the
removal of oil from a large area of oil contaminated soil in Kuwait, due to the
destruction of oil wells during the Iraqi invasion of Kuwait. Chapter Five
focuses on the structural types and biological studies of saponins of I. asprella,
and briefly discusses the characteristics of these chemical structures.
Chapter 1 - Saponins are a group of secondary metabolites characterized
by their strong foam properties in aqueous solution. These compounds are
found in a great number of plant species in many geographical regions and
different climatic zones around the world. Saponins can be classified as
triterpenoid (C30) or steroidal (C27), based on their carbon nucleus
(aglycone). Sugar residues are linked to the aglycone, conferring an
amphiphilic nature on these molecules. The triterpenoid and steroidal saponins
have a different distribution in the plant kingdom. Triterpenoids generally
occur in Leguminosae, Hippocastanaceae, Ranunculaceae, Symplocaceae,
Euphorbiaceae, Verbenaceae and Araliaceae families detected in soybeans,
beans, peas, tea, spinach, sugar beet and quinoa. The major steroidal saponins
viii Candice Greene
are biosynthesized by monocotyledons, such as members of the Liliaceae,

Dioscoraceae, Asparagaceae and Agavaceae families and they are found in
oats, capsicum peppers, aubergine, tomato seed, leek, onion, garlic and
asparagus. The quantification of plant saponins is usually carried out by
spectrophotometric (total saponin content) and chromatographic methods
(specific saponin compound). The most common chromatographic methods
employed are high performance liquid chromatography (HPLC) and ultra
pressure liquid chromatography (UPLC). This technique, coupled to mass
spectrometry, has been used to provide chemical characterization and
composition. The mass analyzers used are a single quadrupole or triple
quadrupole, ion trap and mainly quadrupole time-of-flight (Q-TOF). These
compounds have important structural diversity due to their sugar chains and
substituents, which are relevant to their biological activities. Many in vitro and
in vivo activities have been described for saponins. Triterpenoid saponins have
been shown to display hypocholesterolemic, antispasmodic, antiulcer,
immunomodulatory, antitumoral, antiviral, antibacterial and antifungal
activities, among others. Some of these activities are shared with steroidal
saponins, such as antitumoral, antiviral and antibacterial. In vitro studies with
triterpenoid saponins have also demonstrated hepatoprotective activities.
Furthermore, in vivo studies revealed that saponins exert antinociceptive, anti-
inflammatory and antioxidant activities. Recent studies hold great promise by
saponins in attenuating neurodegenerative diseases and neural trauma,
pointing to their neuroprotective effect, due to diverse mechanisms, such as
anti-oxidation modulation of neurotransmitters, anti-apoptosis, anti-
inflammation, attenuation of calcium influx, modulation of neurotrophic
factors, inhibition of tau phosphorylation and regeneration of neural networks.
Saponins also represent molecules with significant adjuvant activity on the
humoral and cellular immune responses, pointing out their possible use in
vaccine formulations.
Chapter 2 - Obesity predisposes individuals to many metabolic disorders
such as diabetes, cardiovascular diseases, and various types of cancer; it
shortens life expectancy and causes economic losses. Intensive studies have
been conducted to investigate the beneficial effects of numerous food
materials which potentially control obesity. Such foods include ginseng, which
has shown promising results. Ginsenosides are the active biological
compounds in ginseng, and this review focuses on the anti-obesity effect of
ginsenoside Rg3, which, it is claimed, is responsible for the beneficial
properties of ginseng.
Preface ix
Chapter 3 - Saponins represent glycosides, secondary plant metabolites.

Their diverse physicochemical properties have been successfully used in a
number of commercial applications in the food, cosmetics, agricultural and
pharmaceutical sectors. Recently, saponins have become more and more
attractive in environmental applications. Due to their amphiphilic nature and
natural origin, they can behave as biosurfactants. Saponins are able to foam
and to effectively decrease surface/interfacial tension. These properties can be
used in remediation projects to remove organic and inorganic pollutants from
soil. The present chapter presents chemical structure of saponins, their main
sources and surface active properties, including foam production and its
stability, wetting ability, surface tension and critical micelle concentration
(CMC). Based on recent literature, examples of using saponins from different
sources to remove metals from soils, sludges and fly ashes are presented. For
metal removal, saponins can be used in form of solution, foam or
microbubbles. The advantage of using saponins in soil remediation is also their
ability to simultaneously remove combined pollutants, e.g., heavy metals and
polichlorinated biphenyls (PCBs) or polycyclic aromatic hydrocarbons
(PAHs). A promising alternative for remediation of these pollutants can be a
mixing of saponin with chelating agents. Despite the high efficiency of
saponins to remove pollutants from soil, they are still expensive agents. This
chapter demonstrates methods of saponins recovery after soil washing and
attempts of their reuse. A new trends of using saponins in soil remediation are
also presented.
Chapter 4 - The aim of this research was to evaluate the feasibility of
using saponin from soapnuts (Sapindus mukorrossi) for the removal of oil
from a large area of oil contaminated soil in Kuwait which is an aftermath of
the destruction of oil wells during the Iraqi invasion of Kuwait. Experiments
were carried out to assess the efficiency of soil washing using a non ionic
biosurfactant saponin under different conditions like concentration of saponin,
washing time, stirring speed, operating temperature and mass/volume ratio.
The optimum concentration of saponin was found for the most effective
removal of oil from the contaminated sand by using artificial sea water
keeping parameters such as a temperature, soil/solution ratio, stirrer speed and
washing time constant. The results from this study indicates that saponin can
be appropriately used for its application as a biosurfactant for washing oil
contaminated sand.
Chapter 5 - The plant Ilex asprella (Hook. et Arn.) Champ. ex Benth.
(Aquifoliaceae) is a deciduous shrub growing in southern China and Southeast
Asia. The roots of I. asprella, known as Gangmei in China, have been widely
x Candice Greene
used as a Traditional Chinese Medicine (TCM) for the treatment of headaches,

coughs, fever, diarrhea and sore throats. In addition, its root is key ingredient
of ‘Wang-Lao-Ji Herbal Tea’ which was formulated in 1828 and has been
widely accepted as a healthy beverage. Previous investigations on the
phytochemistry of this plant had led to the isolation of triterpenoids,
flavonoids, phenolic acid and alkaloid phenolic compounds. The most
characteristic constituents of I. asprella are triterpenoids and their saponins,
including some rare sulfur-containing derivatives. Pharmacological research
has identified the important associations of I. asprella with their anti-cancer,
anti-inflammatory, and anti-virus activities. This chapter will focus on the
structural types and biological studies of saponins of I. asprella, and the
characteristics of these chemical structures will also be briefly discussed.
In: Saponins: Types, Sources and Research ISBN: 978-1-53610-289-5
Editor: Candice Greene © 2016 Nova Science Publishers, Inc.
Chapter 1
SAPONINS: OCCURRENCE IN NATURE

AND BIOLOGICAL ACTIVITIES
Andresa Heemann Betti, PhD, Juliane Deise Fleck, PhD,

and Simone Gasparin Verza*, PhD
Institute of Health Sciences, Feevale University, Novo Hamburgo,
Rio Grande do Sul, Brazil
ABSTRACT
Saponins are a group of secondary metabolites characterized by their
strong foam properties in aqueous solution. These compounds are found
in a great number of plant species in many geographical regions and
different climatic zones around the world. Saponins can be classified as
triterpenoid (C30) or steroidal (C27), based on their carbon nucleus
(aglycone). Sugar residues are linked to the aglycone, conferring an
amphiphilic nature on these molecules. The triterpenoid and steroidal
saponins have a different distribution in the plant kingdom. Triterpenoids
generally occur in Leguminosae, Hippocastanaceae, Ranunculaceae,
Symplocaceae, Euphorbiaceae, Verbenaceae and Araliaceae families
detected in soybeans, beans, peas, tea, spinach, sugar beet and quinoa.
The major steroidal saponins are biosynthesized by monocotyledons,
such as members of the Liliaceae, Dioscoraceae, Asparagaceae and
Agavaceae families and they are found in oats, capsicum peppers,
*
Corresponding Author: Email: simofar@gmail.com.
2 A. Heemann Betti, J. Deise Fleck and S. Gasparin Verza
aubergine, tomato seed, leek, onion, garlic and asparagus. The

quantification of plant saponins is usually carried out by
spectrophotometric (total saponin content) and chromatographic methods
(specific saponin compound). The most common chromatographic
methods employed are high performance liquid chromatography (HPLC)
and ultra pressure liquid chromatography (UPLC). This technique,
coupled to mass spectrometry, has been used to provide chemical
characterization and composition. The mass analyzers used are a single
quadrupole or triple quadrupole, ion trap and mainly quadrupole time-of-
flight (Q-TOF). These compounds have important structural diversity due
to their sugar chains and substituents, which are relevant to their
biological activities. Many in vitro and in vivo activities have been
described for saponins. Triterpenoid saponins have been shown to display
hypocholesterolemic, antispasmodic, antiulcer, immunomodulatory,
antitumoral, antiviral, antibacterial and antifungal activities, among
others. Some of these activities are shared with steroidal saponins, such
as antitumoral, antiviral and antibacterial. In vitro studies with
triterpenoid saponins have also demonstrated hepatoprotective activities.
Furthermore, in vivo studies revealed that saponins exert antinociceptive,
anti-inflammatory and antioxidant activities. Recent studies hold great
promise by saponins in attenuating neurodegenerative diseases and neural
trauma, pointing to their neuroprotective effect, due to diverse
mechanisms, such as anti-oxidation modulation of neurotransmitters,
anti-apoptosis, anti-inflammation, attenuation of calcium influx,
modulation of neurotrophic factors, inhibition of tau phosphorylation and
regeneration of neural networks. Saponins also represent molecules with
significant adjuvant activity on the humoral and cellular immune
responses, pointing out their possible use in vaccine formulations.
Keywords: triterpenoid saponins, steroidal saponins, activity in vitro, activity

in vivo
1. INTRODUCTION
Saponins are surfactive compounds that are widely distributed in nature,
occurring primarily in plant kingdom (Sparg et al., 2004; Vinken et al., 2007;
Augustin et al., 2011). Plants that produce saponins generally accumulate
these metabolites as part of their development, which can be influenced by
several environmental factors, such as nutrient and water availability, light
irradiation and/or a combination of these factors. The distribution and levels of
Saponins: Occurrence in Nature and Biological Activities 3
saponins have been attributed to different needs of protection against

herbivores and pests (Augustin et al., 2011; Cheok et al., 2014).
The name ‘saponin’ is derived from the Latin word sapo, which means
‘soap’ because saponins form foams when shaken with water (Oleszek, 2002;
Vinken et al., 2007). The ability to form foams is connected with the
amphiphilic structure of saponins, represented by lipophilic sapogenin, named
aglycone, and hydrophilic saccharide side chains (Hostettmann and Marston,
1995; Oleszek, 2002; Augustin et al., 2011).
The saponins can be classified, in accordance with their aglycone
chemical structure, in steroidal saponins (Figure 1), present almost exclusively
in monocotyledonous angiosperms, and in triterpenic saponins (Figure 2),
most common in dicotyledonous angiosperms (Sparg et al., 2004; Vinken et
al., 2007; Cheok et al., 2014). The aglycone derived from a lipophilic steroid
consists of a C27 spirostane skeleton, generally constituted by a six-ring
structure. Triterpenoid aglycone comprises a C30 skeleton consisted of a
pentacyclic structure (Sparg et al., 2004; Augustin et al., 2011; Cheok et al.,
2014).
Saponins are also classified based on the number of attached saccharide
side chains. When only one position of aglycone is glycosylated the saponins
are known as monodesmosidic, whereas when two saccharides side chains are
attached on aglycone the saponins are named bidesmosidic (Hostettmann and
Marston, 1995; Vinken et al., 2007; Augustin et al., 2011). The saccharide side
chain in monodesmosidic saponins is attached at C3 or at C17 position. The
second sugar side chain, in bidesmosidic saponins, is located at C28 (Vinken
et al., 2007; Augustin et al., 2011).
Figure 1. Steroidal aglycones commonly found in plants: (A) spirostane, (B) furostane
and (C) cholestane-type.
Figure 2. Triterpenic aglycones frequently found in plants: (A) β-amyrin (oleanane-

type structure), (B) α-amyrin (ursane-type structure) and (C) lupeol-type.
The saponins generally occur in plants as a mixture of diverse compounds

with very similar physical and chemical characteristics, which make their
separation difficult (Oleszek and Bialy, 2006). Before separation and
identification, saponins can be extracted from vegetal material. To extract
bioactive saponins, conventional extraction techniques are used, such as
maceration, soxhlet, and reflux. Hot extraction may disintegrate some
compounds or generate artefacts. New technologies, like ultrasound-assisted,
microwave-assisted and accelerated solvent extraction have been used more
recently (Oleszek and Bialy, 2006; Cheok et al., 2014).
Several non-chromatographic and chromatographic methods have been
used to identify and quantify saponins in plants. With respect to non-
chromatographic techniques, spectrophotometric techniques like
immunoassays with monoclonal antibodies (MAbs), enzyme-linked
immunosorbent assay (ELISA) or other spectrophotometric methods can be
cited. Regarding chromatographic techniques, thin-layer chromatography
(TLC), low-pressure column chromatography (LPCC), high-performance
liquid chromatography (HPLC) and more recently liquid chromatography
coupled to mass spectrometry (LC-MS) have been used to determinate plant
saponins (Oleszek, 2002; Oleszek and Bialy, 2006; Cheok et al., 2014).
Recently, UPLC coupled with electrospray-ionization quadrupole time-of-

flight mass spectrometry (UPLC-ESI-QTOF-MS) (Görög, 2015) was used to
identify saponins. The difference in the quantitative expressions of saponins
considering spectrophotometric and chromatographic methods is that the last
one allows the quantification of a specific saponin compound while
spectrophotometric methods only express saponins’ total content (Oleszek and
Bialy, 2006).
Due to their chemical properties and abilities as foaming agents, saponins
or plant that contains saponins have been used by industries as additives to
foods and cosmetics. Saponins have a diversity of biological and
pharmaceutical properties, such as antimicrobial, antifungal, antiviral,
haemolytic, molluscicidal, anti-inflammatory, cytotoxic and antitumor
activities, hipocholesterolemic, expectorant and immunoadjuvant activities.
Considering the importance of this class of substances the aim of this chapter
is to review the most important biological properties of saponins and provide
an overview on the current knowledge about it.
2. IMMUNOLOGICAL ADJUVANT AND

IMMUNOMODULATORY PROPERTIES
Adjuvants are compounds that improve the immune response to an
antigen. The incorporation of adjuvants into inoculated antigen formulations is
used to different purposes (Aguilar and Rodríguez, 2007; Mount et al., 2013;
Brito and O'Hagan; 2014):
a) to enhance the immunogenicity of highly purified or recombinant

antigens;
b) to reduce the amount of antigens or the number of immunizations
needed for protective immunity;
c) to improve the efficacy of vaccines in newborns, elderly or immune-
compromised people; or
d) as antigen delivery systems for the uptake of antigens by the mucosa.
The adjuvant type can affect the nature of the immune responses, and
these molecules can lean the immune system in favor of Th1 or Th2 type
responses. Th1 immune response is a requisite for cytotoxic T lymphocyte
(CTL) production. This response is characterized by the production of
cytokines IL-2, TNF and interferon gamma (INF-), and an enhanced

production of IgG2a, IgG2b and IgG3 in mice. Th2 response is represented by
the production of cytokines IL-4, IL-5 and IL-10, as well as an enhanced
production of IgG1. Immunity to infectious agents requires distinct types of
immune responses. For example, Th1 response is required for protective
immunity against intracellular infectious agents, such as viruses, certain
bacteria and protozoa, and presumably against cancer cells, whereas Th2
immunity is effective for protection against most bacteria as well as certain
viral infections (McKee et al., 2007; Mount et al., 2013; Brito and O'Hagan;
2014).
Saponins is able to activate the mammalian immune system, which has led
to significant interest in their potential as vaccine adjuvants. They can
stimulate both Th1 (predominantly) and Th2 responses. For instance, the
capacity of Quillaja saponaria saponins to stimulate Th1 immune response
and the production of CTLs against exogenous antigens makes them ideal for
using in subunit vaccines and vaccines directed against intracellular pathogens,
as well as for therapeutic cancer vaccines (Skene and Sutton, 2006; Sun et al.,
2009; Petrovsky, 2015).
Q. saponaria Molina (Quillajaceae), a native species from South America,
is the major source of saponins with immunoadjuvant activity in vaccines,
which was demonstrated in numerous studies (Chapman et al., 2000; Evans et
al., 2001; Stoute et al.; 2007; Xiao et al., 2007; Vandepapelière et al., 2008;
Wang et al.; 2008; Spilki et al., 2010; Garçon and Van Mechelen, 2011; The
RTS, 2011; Dendouga et al., 2012; The RTS, 2014; Kaur et al., 2015; Lal et
al., 2015; Leroux-Roels et al., 2015; Martinez et al., 2015; Marty-Roix et al.,
2016; Thakur et al., 2015; Lage et al., 2016; Ng et al., 2016; Tafaghodi et al.,
2016). The first enriched mixture of saponins from Q. saponaria bark, named
Quil-A®, was obtained by Dalsgaard in the 1970s. Quil-A® has been used in
experimental vaccines as well as veterinary commercial vaccines. However, it
is unsuitable for human vaccines due to its toxicity. In 1991, Kensil et al.
isolated and characterized four major saponins from aqueous extract of Q.
saponaria bark (named QS-7, QS-17, QS-18 and QS-21), which showed
similar adjuvant properties, but differed considerably in their toxicity. QS-21
(Figure 3) proved to be a potent adjuvant with an acceptable toxicity level,
utilized in several subsequent studies, including clinical trials (Chapman et al.,
2000; Evans et al., 2001; Stoute et al.; 2007; Vandepapelière et al., 2008;
Wang et al.; 2008; Garçon and Van Mechelen, 2011; The RTS, 2011; The
RTS, 2014; Lal et al., 2015; Leroux-Roels et al., 2015).
Figure 3. QS-21 structure.
The saponins from Q. saponaria are glycosylated triterpenes, classified as

bidesmosides. The major aglycone in these saponins is quillaic acid (3, 16-
dihydroxy-23-oxolean-12-en-28-oic acid), which is characterized by an
aldehyde group attached to position 4 (van Setten, and van de Werken, 1996).
Besides quillaic acid, other aglycones have been identified, such as 22-
hydroxy-quillaic acid, phytolaccagenic acid and echinocystic acid (Guo and
Kenne, 2000). The basic structure reported for the major saponin of Q.
saponaria is a quillaic acid, substituted in position 3 with a di- or trisaccharide,
and in position 28 with an oligosaccharide linked through a fucose residue to
which one or two acyl groups are linked (Nord and Kenne, 2001).
The immunological adjuvant activity of saponins from Q. saponaria is also
observed when incorporated into lipid-based delivery systems, such as
immune stimulating complexes (ISCOMs), emulsion droplets and liposomes.
These formulations were able to enhance antigen presentation by antigen-
presenting cells (APCs) and induce predominantly cytotoxic T-lymphocyte
(CTL) production, eliciting both Th1 and Th2 cytokine secretion. ISCOMs are
particulate antigen delivery systems composed by saponins, cholesterol,
phospholipids, and immunogens. When the particulate system is formulated
without immunogens, they are called ISCOMATRIX. Different antigens have
been used to form ISCOMs, including the derived from herpes simplex virus
type 1 (HSV-1), human immunodeficiency virus (HIV-1), cytomegalovirus
(CMV), Ebola virus (EBOV), Epstein–Barr virus (EBV), hepatitis B virus
(HBV), rabies virus, influenza viruses and Leishmania spp. (Barr and
Sjölander, 1998; Coulter et al., 2003; Kim et al., 2006; Skene et al., 2008;
Pedersen et al., 2011; Magnusson et al., 2014; Martinez et al., 2015;
Mehravaran et al., 2016; Bengtsson et al., 2016). Bigaeva et al. (2016) revised
the safety and tolerability of QS-21 and ISCOMATRIX-adjuvanted vaccines.
The authors analyzed nine randomized controlled trials, and suggest that the
use of ISCOMATRIX enables a better systemic tolerability profile when
compared to the use of QS-21, however, no better local tolerance was
observed in immunized non-healthy subjects.
Liposome-based adjuvants are highly versatile, as they can be tailored
through (Schmidt et al., 2016):
a) the choice of lipid composition,

b) the inclusion of immunostimulating compounds,
c) the choice of formulation method, and
d) the mode of antigen and immunostimulator association.
The liposomal adjuvant system AS01 (containing MPL and QS21) has
showed, in clinical trials, significantly adjuvant proprieties in vaccines against
malaria, HIV, M. tuberculosis, HBV, and Herpes Zooster (The RTS, 2011;
Garçon and Van Mechelen, 2011; Didierlaurent et al., 2014; The RTS, 2014;
Davitt and Lavelle, 2015; Lal et al., 2015; Leroux-Roels et al., 2015; Schmidt
et al., 2016).
Other sources from immunoadjuvant saponins have been evaluated,
amongst them Quillaja brasiliensis, Panax spp., Platycodon grandiflorus,
Chenopodium quinoa, Inga laurina, and Glycine max. The saponins from Q.
brasiliensis (Quillajaceae), native species from southern Brazil and Uruguay,
are remarkably similar to those of Q. saponaria (Kauffmann et al., 2004).
Activity comparable to that of commercial saponin-adjuvant Quil-A® was
reported for the aqueous extract (AE) and a purified saponin fraction (QB-90)
obtained from Q. brasiliensis leaves. These studies described vaccines against
bovine herpes virus type 1 and 5, human poliovirus, bovine viral diarrhea virus
and rabies virus in mice (Fleck et al., 2006; Silveira et al., 2011; de Costa et
al., 2014; Cibulski et al., 2016; Yendo et al., 2016).
Ginsenosides, saponins from P. ginseng and P. notoginseng (Araliaceae),
significantly enhanced a specific antibody and cellular response when
employed as vaccine adjuvants against ovalbumin (OVA) and Candida
albicans in mice (Sun et al., 2004; Sun et al., 2008; Yang et al., 2007; Han and
Rhew, 2013). Ginsenosides also showed immunomodulatory activity in
healthy volunteers, evidenced by improvements in PMN cell chemotaxis,

phagocytosis, and total number of T-helper and T-suppressor cells. These
saponins showed opposing relationships between the adjuvant and haemolytic
activities (Sun et al., 2005; Sun et al., 2006; Kumar et al., 2012).
Platycodon grandiflorus (Campanulaceae) main bioactive compounds are
oleanane-type saponins. These saponins have shown adjuvant effect against
OVA in mice, with efficacy on Th1 and Th2 immune response (Xie et al.,
2008a; Xie et al., 2008b; Xie et al., 2008c). Platycodin D has been evaluated,
in mice, as adjuvant in vaccine formulations to recombinant hepatitis B surface
antigen, and Newcastle disease virus-based recombinant avian influenza
vaccine (Xie et al., 2009; Xie et al., 2010). Different from what was found in
the ginsenosides, Sun et al. (2011) observed a tendency in the platycodigenin-
type saponins with higher adjuvant potentials of showing higher haemolytic
activities.
In addition, saponins from Chenopodium quinoa (Amaranthaceae) have
been employed to enhance immune responses and mucosal responses after
intragastric and intranasal coadministration with either cholera toxin or OVA
(Estrada et al., 1998). Later, Verza et al. (2012) showed that quinoa saponins
fractions significantly enhanced the production of humoral and cellular
immune responses to OVA in mice.
Cruz et al. (2016) described the purification and isolation of ingasaponin,
a complex triterpenoid saponin from Inga laurina (Leguminosae). Ingasaponin
showed adjuvant activity on the cellular immune response against OVA
antigen, in mice. Immunologic adjuvant properties against OVA is also
described to soyasaponins, a group of complex and structurally diverse
oleanane triterpenoids occurring mainly in Glycine max (Oda et al., 2000; Oda
et al., 2003; Sun et al., 2014).
3. ACTIVITIES ON MEMBRANES
Due to their chemical composition, saponins are considered to be part of
the defense systems of plants. Several reports highlight antimicrobial (Sparg et
al., 2011; El Dib et al., 2015; Sidana et al., 2016), fungicidal (Sidana et al.,
2016), insecticidal, molluscicidal, haemolytic and citotoxic (Wu et al., 2015)
activities of saponins. The molecular mechanism involved in these functions
are not completely described but the ability to cause membrane perturbation is
a property of saponins that can be related to the others listed before (Augustin
et al., 2011).
3.1. Haemolytic Activity
Saponins have the ability to lyse erythrocytes (Sparg et al., 2004). This
property was first described by Dourmashkin et al. (1962), who observed
formation of pores in the viral membrane coat as a consequence of saponin
treatment. The haemolytic property is related with characteristics of chemical
structure of aglycone (Oda et al., 2000), number and length of saccharide side
chains and substituents in saccharide chains (Woldemichael and Wink, 2001).
Generally, saponins possess powerful haemolytic activity due to their higher
affinities for cholesterol on erythrocyte membrane (Oda et al., 2000; Silva and
Parente, 2013). A high number of studies attemped to find a relationship
between saponins structure and their membrane perturbation activity (Wang et
al., 2007; Gauthier et al., 2009; Sun et al., 2011; Liu et al., 2013), however
these studies are conflicting in their results (Augustin et al., 2011).
3.2. Antifungal Activity
The antifungal/antiyeast activity has been reported since 1993, when

Othani et al. has identified antifungal saponins obtained from Rapanea
melanophloeos leaves. In 1998, Sindambiwe et al. reported antifungal activity
for saponins from Maesa lanceolata. The authors reported that a mixture of
saponins inhibited the growth of Epidermophytum floccosum, Microides
interdigitalis, Tricophyton rubrum, Candida albicans and Microsporum canis.
Another plant species that is worth mentioning by its antifungal activity is P.
notoginseng. Ma et al. (1999) tested fourteen saponins and seven of them
demonstrated inhibitory effect on Aphanomyces cochlioides zoospore motility.
Saponins from Columbrina retusa, named jujubogenin saponins, showed
activity against Candida albicans, Cryptococcus neoformans and Aspergillus
fumigatus (Li et al., 1999). Monodesmosidic saponins isolated from Hedera
colchica exhibited antifungal activity, however it was lower than observed for
reference antifungal agents. The authors have demonstrated that the aglycone
and the number, kind and sequence of sugar chains had a significant influence
on antifungal activity (Mshvildadze et al., 2000).
Escalante et al. (2002) isolated the saponins of Phytolacca tetramera and
tested their antifungal activity against several fungal strains. The most
sensitive fungus was Tricophyton mentagrophytes. Triterpenoid saponins from
Chenopodium quinoa have been reported by their activity against Candida
albicans (Woldemichael and Wink, 2001) and Botrytis cinerea (Stuardo and
San Martin, 2008), a fungus that is the causal agent of gray mold disease.
Chapagain et al. (2007) have demonstrated the antifungal activity of
saponin-rich extracts from Balanites aegyptiaca, Quillaja saponaria and
Yucca schidingera, tested against common phytopatogenic fungi (Pythium
ultimum, Fusarium oxysporum, Alternaria solani, Colletotrichum coccodes,
and Verticillium dahliae). The results showed a specific and dose-dependent
saponin antifungal activity.
Saponins from Allium species have also been reported by their antifungal
activity (Barile et al., 2007; Lanzotti et al., 2012). A. minutiflorum showed
significant activity when tested against soil-borne pathogens (Fusarium
oxysporum, F. oxysporum, F. solani, P. ultimum and Rhizoctonia solani) and
air-borne pathogens (Botrytis cinerea, Alternaria alternata and A. porri)
(Barile et al., 2007). Antifungal activity of seven isolated garlic saponins
(furostane and spirostane saponins) was tested against Trichoderma harzianum
and Botrytis cinerea. The antimicrobial effect of the isolated compounds was
concentration-dependent. Furthermore, while all seven compounds showed a
significant activity against T. harzianum, only five were effective against B.
cinerea (Lanzotti et al., 2012).
Antifungal activity against Candida albicans, C. glabrata, C. krusei and
Cryptococcus neoformans was also reported for furostane and spirostane
glycosides of Agave spp. (Sidana et al., 2016). In another study, Ribeiro et al.
(2013) evaluated saponins from sisal (Agave sisalana) and juá (Ziziphys
joazeiro). While saponins obtained from sisal possessed no antimicrobial
activity, juá saponins presented antifungal activity against Candida albicans.
3.3. Antimicrobial Activity
Antimicrobial activity has been reported for triterpenoid saponins since

1988 (Killen et al., 1988; Mahato et al., 1988). Saponins isolated from alfafa
and derived from oleanolic acid had activity against Cryptococcus neoformans
(Mahato et al., 1988). The same authors reported antimicrobial activity for
saponins derived from korean ginseng, Anagallis arvensis, Phytolacca
octandra and Camellia sp.
Triterpenoid saponins derived from Hedyotis nudicaulis were tested
against Bacillus subtilis and demonstrated weak antibacterial activity. Results
indicated that the number of sugar chains exert an influence in antimicrobial
activity (Konishi et al., 1998). Iorizzi et al. (2002) studied the furostane
saponins from Capsicum annuum seeds. They showed weak or no growth

inhibition against Gram-positive and Gram-negative bacterias.
Antimicrobial properties of several crude extracts and pure saponins from
the aerial parts of Astragalus verrucosus were investigated by the disc
diffusion method. All extracts tested were inactive against the randomly
selected Gram-negative bacterias. Only the n-hexane extract was active against
the Gram-positive bacteria, Staphylococcus aureus (Pistelli et al., 2002).
Antimicrobial properties against Bacillus megaterium, Salmonella
Typhimurium and Pseudomonas aeruginosa were reported for saponins
derived from Acacia auriculoformis. The saponins acasiaside A and B
demonstrated complete inhibition of the cited microrganisms at the
concentration of 700 µg/mL or higher (Mandal et al., 2005).
Drypete inaequalis is a forest shrub used for the treatment of sinusitis,
swellings, boils, gonorrhea and dysentery. The saponins present in this species
were identified as serjanic acid, oleanolic acid, hederagenin, keretaroic acid,
stigmasterol and sitosterol. Four triterpene derivatives were isolated and the
antimicrobial activity was tested using the agar diffusion method. The
triterpenes were tested against some Gram-positive and Gram-negative
bacterias, demonstrating weak activity (Awanchiri et al., 2009). On the other
hand, a triterpenoid saponin derived from D. laciniata exhibited moderate
activity against the microorganisms tested (Fannang et al., 2011).
Hassan et al. (2010) studied saponin-rich extracts from guar meal,
evaluating the haemolytic and antimicrobial activities. Antibacterial activity
was evaluated against S. aureus, Lactobacillus spp., Typhimurium and E. coli.
Only the 100% methanol fraction demonstrated both haemolytic and
antibacterial activities against the microorganisms tested (Hassan et al., 2010).
Fouedjou et al. (2014) reported the chemical structure of three new
steroidal saponins from Cordyline fructiosa. The authors evaluated cytotoxic
and antimicrobial activities of these saponins and only one saponin showed
moderate antibacterial activity against Gram-positive Enterococcus faecalis.
Two of the three saponins tested demonstrated moderate cytotoxic activity
against MDA-MB 231, an adenocarcinoma cell line.
More recently, Scalia et al. (2015) studied the saponins of Luffa
operculata against S. aureus, Streptococcus pneumoniae and Streptococcus
pyogenes. It was evaluated the antimicrobial activity of an alcoholic extract of
L. operculata diluted at the concentrations of 0.2% to 2.0% in TSB culture
medium. At the concentration of 2.0%, L. operculata extract was active
against all bacterias tested. Since a number of substances are present in L.
operculata extract, such as curcubitacines B and D, glycosides, free sterols,
organic acids, phenols and saponins, the antimicrobial activity could not be
attributed exclusively to the saponins in the extract.
3.4. Antiviral Activity
Saponins have also been reported by their antiviral activity since 1987 by
Amoros and Guirre for triterpene saponins from Anagallis arvensis. The
authors isolated two saponins and tested antiviral activity against Herpes
simplex virus type 1 and poliovirus. The antiviral compounds had
demonstrated highly haemolytic activity. Triterpenoid saponins isolated from
the leaves of Maesa lanceolata were tested against HSV-1 and HIV viruses.
The authors concluded that a free OH at 16 position and acylation of the 22-
OH appears to be essential for HSV-1 activity. However, the same saponins
have no activity against HIV virus (Apers et al., 2001).
Saponins from Ilex spp. have been studied by their antiviral activity. Wu
et al. (2007) isolated six triterpenoid saponins and tested their antiviral
activity. A saponin designated as oblonganoside A showed an inhibitory
activity against Tobacco mosaic virus when evaluated in vitro.
The Tibetan herb Potentilla anserine L. has been widely used in China to
treat hepatitis B. Zhao et al. (2008) isolated a triterpenoid from P. anserine, an
ursane –type saponin and evaluated the inhibitory activity of this compound on
HBV in vitro. The results showed that the isolated saponin could decrease the
expression levels of HBsAg, HbeAg and HBVDNA. In addition, antiviral
activity was not correlated with saponin cytotoxicity.
A methanolic extract of powered roots of Achyranthes aspera and an
isolated saponin (oleanolic acid) were evaluated against herpes simplex virus
type-1 (HSV-1) and type-2 (HSV-2). The phytochemical screening of
methanolic extract of A. aspera showed the presence of alkaloids,
carbohydrates, glycoproteins, sterols, triterpenes and flavonoids. Methanolic
extract evidenced weak anti-herpes virus activity, while oleanolic acid
exhibited potent antiherpesvirus activity against HSV-1 and HSV-2. These
results suggest that the antiviral activity attributed to A. aspera extract could
be due to saponins (Mukherjee et al., 2013).
Nyakudya et al. (2014) studied Platycodon grandiflorum, which contains
a mixture of chemical compounds whose main bioactive components are
saponins named platycosides. A pharmaceutical composition from this species
roots was developed for the prevention or treatment of hepatitis C.
Antiviral activity of ginsenosides, saponins from ginseng, was evaluated.

Seven ginsenosides (Re, Rf, Rg2, Rb1, Rb2, Rc and Rd) were tested against
coxsackievirus B3 (CVB3), enterovirus 71 (EV71) and human rhinovirus 3
(HRV3). The antiviral assay demonstrated that protopanaxatriol type
ginsenosides (Re, Rf and Rg2) have significant antiviral activities against
CVB3 and HRV3. Only Rg2 showed anti-EV71 activity. The protopanaxadiol-
type ginsenosides (Rb1, Rb2, Rc and Rd) did not showed any antiviral activity
against CVB3, EV71 and HRV3 (Song et al., 2014). Additionally, antiviral
properties of korean red ginseng (KRG), a heat-processed ginseng, developed
by the repeated steaming and air-drying of fresh ginseng were studied. Im et
al. (2015) reported that KRG possess protective effects against infections with
human pathogenic viruses such as respiratory syncytial virus, rhinovirus,
influenza virus, human immunodeficiency virus, human herpes virus, hepatitis
virus, norovirus, rotavirus, enterovirus and cosxackevirus.
Hedera helix leaf extract has been used to treat inflammatory bronchial
diseases. Several saponins were isolated from H. helix including α-hederin,
hederasaponin-C, hederacolchiside-E and hederacolchiside-F. Hong et al.
(2015) investigated antiviral properties against influenza A/PR/8 (PR8) virus
in mice. Compared to oseltamivir treatment alone, the coadministration of H.
helix extract containing the highest proportion of hederasaponin F decreased
pulmonary inflammation in PR8-infected mice (Hong et al., 2015).
Bupleurum spp. roots (radix blupleuri) are frequently used as herbal
treatments for liver diseases in Eastern Asia. Their pharmacology and
chemical composition have been extensively studied and the bioactive
compounds are saponins, named saikosaponins. Recently, Lin et al. (2015) has
demonstrated that a methanolic extract from Blupleurum kaoi roots and its
purified saikosaponins (SSa, SSb2, SSc and SSd) inhibited HCV infection.
This study identified SSb2 as the most effective anti-HCV saponin.
3.5. Hypocholesterolemic Activity
Saponins have been studied due to their lowering cholesterol effects, an

activity that has been attributed to the increase of fecal cholesterol excretion,
enhanced conversion of cholesterol to bile acids in liver and reducing the
plasma cholesterol (Jia et al., 2010; Resende et al., 2012; Elekofehinti et al.,
2013; Dooren et al., 2015; Liu et al., 2016). The liver plays an important role
in maintaining cholesterol homeostasis, eliminating cholesterol from the body
via bile by converting cholesterol into bile acids or directly secreting biliary
cholesterol. Other mechanisms involved in the modulation of the expression of

hepatic and intestine genes are related to cholesterol metabolism (Jia et al.,
2010; Resende et al., 2012; Elekofehinti et al., 2013; Dooren et al., 2015; Liu
et al., 2016). Studies also suggested that saponins partially suppressed
intestinal absorption of cholesterol by inhibiting micellar solubilization in the
small intestinal lumen and that saponins disrupt cholesterol micelle formation
(Ramirez-Jimenez et al., 2015; Liu et al., 2016).
Some important species reported with hypocholesterolemic activities are
P. notoginseng (Burk.) F.H. Chen (Jia et al., 2010), Ilex paraguariensis A. St.
Hill (Resende et al., 2012), Solanum anguivi Lam. (Elekofehinti et al., 2013)
Herniaria hirsuta L (Dooren et al., 2015) and Gynostemma pentaphyllum (Liu
et al., 2016).
4. ANTIINFLAMATORY AND ANTIOXIDANT ACTIVITIES

Inflammation is an immunological defense mechanism elicited in response
to several noxious stimulus. Inflammation processes are complex and involve
several events such as vasodilatation, plasma extravasations, cellular migration
and release of various mediators, which is usually associated with pain
(Florentino et al., 2016). Saponins have been studied with respect to their
activities involving inflammatory and oxidative processes. In addition, it has
been well documented that saponins suppress oxidative stress in neurons and
neuronal cell lines induced by pro-oxidants due to their antioxidant activity.
Therefore, the most commonly accepted explanation for saponins
neuroprotective effects is related to their anti-inflammatory and antioxidant
activities (Lopez et al., 2007; Qian et al., 2009). Some of these important
biological activities are described below.
4.1. Anti-Inflammatory Activity
Sepsis is a major clinical challenge in modern medicine, representing one

of the leading causes of death in developed countries. The syndrome is a
consequence of a dysregulated immune response, including early uncontrolled
systemic inflammation and prolonged immunosuppression in the late phase.
Liu et al. (2016) studied the effect of astragaloside IV in an animal model of
sepsis. Astragaloside IV significantly improved survival in septic mice. In
agreement with this protective effect, the pathologic damage that was typically
seen in lung and spleen was ameliorated, the level of bacterial burden was
lessened, inflammatory cytokines and chemokines in circulation were reduced,
and lymphocyte apoptosis was inhibited. It also suppressed LPS-induced
macrophage activation through inhibition of NF-κB and ERK1/2 signaling
pathways.
Yang et al. (2014) investigated the anti-inflammatory effect of 21-O-
angeloyltheasapogenol E3, a triterpenoid saponin isolated from the seeds of
the tea tree Camellia sinensis (L.) O. Kuntze. This saponin inhibited
macrophage mediated inflammatory responses such as phagocytic uptake,
ROS generation, and NO production, effects mediated by suppression of
inflammatory pathways composed of AKT, IKK, and NF-𝜅B.
In addition, the Gleditsia sinensis thorns have been used in China and
Korea as traditional medicine for the treatment of some inflammatory diseases
such as swelling, carbuncle, suppuration and skin diseases (Zhang et al.,
2016). Its fruit hull has been used as a traditional anti-inflammatory medicine
for the treatment of some respiratory system diseases caused by inflammation.
Choi et al. (2012) confirmed this anti-inflammatory action in their study. They
verified that the fruit hull inhibited neutrophilic lung inflammation by
activating the anti-inflammatory factor Nrf2. Kim et al. (2014) carried out a
similar assay and provided experimental evidence that the fruit hull suppressed
the LPS-induced lung inflammation in acute lung injury mouse model, at least
partly by the mediation of Nrf2 activation. These findings suggest that the fruit
hull possesses modest inhibitory effect on acute inflammation that can be
mediated by weakening the inflammatory effects of mediators such as
serotonin (Dai et al., 2002).
Xiong et al. (2015) tested the anti-inflammatory effect of different
triterpene saponins from the stems of Entada phaseoloides by evaluating the
activity against NO production in lipopolysaccharide-stimulated mouse
macrophage RAW264.7 cells. They significantly inhibited the levels of
proinflammatory cytokines, such as TNF-α, IL-1β, IL-6 and IL-8.
The compound K (20-O-d-glucopyranosyl-20(S)-protopanaxadiol), a
novel ginsenoside metabolite and member of the dammarane-type triterpene
saponins, was studied by Chen et al. (2016). This compound suppressed
memory B cell subsets and suppressed CD40L expression on T cells and
CD40 expression on B cells, demonstrating that it downregulated memory B
cells in adjuvant-arthritis rats, and this down-regulation may be T-cell
dependent.
Mao et al. (2016) studied the gastroprotective effects of astragaloside IV
from Astragalus membranaceus on acute gastric lesions in rats under stressful
conditions. The treatment with astragaloside IV significantly decreased the

size of gastric lesions and levels of MDA, TNF-α and MCP1, and, in addition,
normalized gastric pH, gastric mucus and SOD levels. Also, the pretreatment
with astragaloside IV resulted in significant reduction in Bax, over-expression
of PLCγ response level and elevation in 70-kDa heat-shock protein (HSP70).
HSP70 is a cytokine that binds with high affinity to the plasma membrane,
causes a rapid intracellular calcium flux, activates NF-κB and upregulates the
expression of pro-inflammatory cytokines tumor necrosis factor (TNF)-α and
interleukin (IL-1β and IL-6) in human monocytes (Asea et al., 2000). The
study demonstrated that the acute gastric lesions induced are attenuated by the
pretreatment with astragaloside IV, which is possibly due to the enhancing of
the expression of HSP70 with concomitant antioxidant, anti-inflammatory and
anti-apoptotic capacities (Mao et al., 2016).
4.2. Antinociceptive Activity
Ahn et al. (2016) evaluated the antinociceptive effect of ginsenoside Rg3,

an extract of total ginseng saponins, which accounts for 4.7% of all saponins,
in a model of rats underwent plantar incision. This study demonstrated an
analgesic effect with a curvilinear dose-response relationship. Alpha 2
adrenergic receptor appeared to be related to the analgesic effect of
ginsenoside Rg3. In addition, this study suggested that the anti-inflammatory
effect of ginsenoside Rg3 could be related to its analgesic effect.
The antinociceptive activity of astragaloside IV was studied by Shi et al.
(2015) in an animal model of chronic constriction injury, used in the treatment
of refractory neuropathic pain. Astragaloside inhibited allodynia and
hyperalgesia induced by mechanic and thermal stimuli, as well as
downregulated the expressions of a series of proteins involved in mediating
neuropathic pain in the dorsal root ganglia. In addition, it restored the nerve
conduction velocity and the histological structure of the damaged sciatic nerve
23 days after the surgery. Results from immunoelectron microscope showed
that glial cell-derived neurotrophic factor (GDNF) family receptor α1 induced
by astragaloside could form a circular band in the myelin debris between the
injured axons and Schwann cells, contributing toward restoration of the
damaged nerve.
Li et al. (2012) investigated the antinociceptive effect of intrathecal escin
from Aesculus hippocastanum by examining its effect on the formalin-induced
activation of c-Fos, and phosphorylated p38 mitogen-activated protein kinase
(p-p38 MAPK) in the rat spinal cord. Escin decreased pain-related behavior in
the rat formalin test and reduced Fos immunoreactivity and p-p38 MAPK
immunoreactivity in the spinal cord dorsal horn after formalin injection,
indicating that escin provided antinociceptive effect in the rat formalin test.
Finally, the effect of saponins associated with other drugs was studied.
Kim et al. (2014) investigated the antinociceptive activity of ginseng total
saponins on hyperalgesia induced by repeated intramuscular injections of
acidic saline in rats. Ginseng total saponins showed an antinociceptive activity
against chronic muscle-induced pain, and their antinociceptive effect on
mechanical hyperalgesia may be mediated by NMDA, suggesting that ginseng
can be useful in the treatment of chronic musculoskeletal pain syndrome.
Aditionally, Yoon et al. (2011) studied the synergistic interaction between
intrathecal ginsenosides and morphine on formalin-induced nociception in
rats. This study demonstrated that the coadministration of ginsenosides and
morphine resulted in a synergistic antinociception mediated by μ, δ, and κ
opioid receptors. Rauf et al. (2011) also investigated the effect of morphine
combined with saponins. Bacopasides, triterpene saponins isolated from
Bacopa monnieri, were studied for acquisition and expression of morphine
tolerance in mice. Their acute and chronic administration significantly reduced
the expression and the development of tolerance to morphine analgesia in
mice. In addition, it enhanced antinociceptive effect of morphine in intolerant
animals. This study suggests the effectiveness of B. monnieri for the
management of morphine tolerance (Rauf et al., 2011).
4.3. Antioxidant Activity
Saponins have a well-documented antioxidant effect in vitro and in vivo.

Ye et al. (2009) studied the antioxidant mechanism of ginsenoside Rd and
indicated that it reduces the intracellular production of reactive oxygen species
(ROS) and malondialdehyde (MDA), increasing glutathione content, and
enhancing the antioxidant enzymatic activities of catalase, superoxide
dismutase (SOD), and GSP-Px.
Notoginsenoside R1, in its turn, protects neurons from excitotoxicity,
increase of intracellular free Ca2+, overproduction of intracellular reactive
oxygen species and depolarization of mitochondrial membrane potential in
cultured neurons exposed to glutamate, in addition to blocking decreased Bcl-
2 and increased Bax expression levels (Gu et al., 2009).
Li et al. (2009) indicated that the neuroprotective effect of total saponins

of P. notoginseng on focal ischemia might be related to inhibition of apoptosis
and caspase activation, since they reduced the expression of caspase-1 and
caspase-3, and they had no effect on the expression of caspase-8.
Luo et al. (2011) studied the effect of garlic saponins in a model of
hypoxia, which occurs when the excessive ROS induced by hypoxia result in
cell injury and dysfunction. In this study, garlic saponins improved cell
viability in PC12 cells, decreased lactate dehydrogenase (LDH) leakage and
caused the cells to maintain neuronal-like characteristics in hypoxia. The
production of MDA and 8-hydroxy-deoxyguanosine (8-OH-dG) was
attenuated by the saponins, indicating its antioxidant properties.
Arjunolic acid, a pentacyclic triterpenoidal saponin of Terminalia arjuna
is also well recognized for its antioxidant properties. Yaidikar and Thakur
(2015) evaluated its antioxidant potential against focal cerebral ischemia
reperfusion injury in rats subjected to middle cerebral artery occlusion.
Arjunolic acid prevented neuronal damage induced by ischemia reperfusion by
regulating the levels of MDA, reduced glutathione (GSH), nitric oxide (NO),
protein carbonyl content and mitochondria generated reactive oxygen species.
It also controlled the enzyme activities of Na+-K+-ATPase, SOD, catalase,
glutathione peroxidase (GPx) and glutathione reductase (GR).
4.4. Neuroprotective Activity
The neurprotective effects of saponins on diseases and injury of the

nervous system have been reported increasingly. Studies have shown that
saponins are effective in attenuating neurodegenerative diseases and neural
trauma. Sun et al. (2015) revised that saponins exert neuroprotective activities
via diverse mechanisms, including antioxidation, modulation of
neurotransmitters, anti-apoptosis, anti-inflammation, attenuation of Ca2+
influx, modulation of neurotrophic factors, inhibition of tau phosphorylation,
and regeneration of neural networks. Aditionally, they regenerate the neural
network by directly promoting cell survival, improving neurite outgrowth, and
recovering the activities of axons and synapses, as well as preventing
malfunction of neurons by altering levels of neurotransmitters,
neurotransmitter receptors, and second messengers, promoting cell survival
(Sun et al., 2015).
Many saponins have been reported for neuroprotection purposes. P.
notoginseng saponins, which has been extensively used in the treatment of
coronary heart disease, ischemic cerebrovascular disease and hemorrhagic

disorders in China over hundreds of years, was studied by Shi et al. (2016).
They provided neuroprotective effects in a rat model of cerebral ischemia and
SH-SY5Y cells exposed to oxygen/glucose deprivation injury by inhibiting the
overexpression of NgR1, RhoA, and ROCK2. Liu et al. (2009) studied
ginsenoside Rd for the treatment of ischemic stroke in a multicenter,
randomized, double blind, and placebo-controlled phase II trial. Ginsenoside
Rd partly improved neurological deficits in patients with acute ischemic stroke
and indicated that there was a significant improvement in the scores of
National Institutes of Health Stroke Scale after 15 days of treatment. Tohda et
al. (2004) demonstrated that ginsenoside Rg1 and Rb1 increased neural
plasticity; also, they increased proliferation and differentiation rates of neural
progenitor cells in dentate gyrus of hippocampus of normal adult mice and
global ischemia model in gerbils. These findings are important for the
treatment of neurodegenerative disorders characterized by neuronal loss, such
as Alzheimer disease (Cheng et al., 2005). Timosaponins, one group of the two
major components of Anemarrhean asphodeloides, play a central role in the
development of Alzheimer disease, since these saponins can enhance the
learning and memory capacities in rats with β-amyloid protein induced
dementia (Ouyang et al., 2005). Ginseng and ginsenoside Rg1, Rg3, and Re
also resulted in significant reductions in the amount of β-amyloid protein
detected in the brains of animals after single oral doses of these agents,
indicating that ginseng and its purified ginsenosides may have similarly useful
effects in human disease (Chen et al., 2006).
Furthermore, panaxatriol saponins extracted from P. notoginseng provided
neuroprotection against the loss of dopaminergic neurons and behavioral
impairment caused by MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine),
used in an experimental model of Parkinson disease, since they increased
tyrosine hydroxylase, suppressing cycloxygenase-2 over expression and
inhibiting mitochondria-mediated apoptosis (Luo et al., 2011). Astragaloside
IV protected dopaminergic neurons against 6-OH-DA (6-hydroxydopamine)-
induced degeneration, promoted neurite outgrowth, and increased tyrosine
hydrolase and nitrite oxide synthase of dopaminergic neurons. This
neuroprotective effect was specific for dopaminergic neurons, suggesting its
potential for Parkinson disease (Chan et al., 2009).
Moreover, evidence indicates that neuronal calcium (Ca2+) signaling is
abnormal in many neurodegenerative disorders, such as Huntington’s disease,
a neurodegenerative genetic disorder with progressive cell death, leading to
movement, cognitive, and psychiatric problems (Jakel and Maragos, 2000; Sun
et al., 2015). Since Ca2+ fluxes across the plasma membrane are essential for
fundamental functions of neurons, synaptic dysfunction, impaired plasticity,
and neuronal degeneration can be resulted from compromised cellular Ca2+-
regulating systems. Wu et al. (2009) demonstrated that ginsenoside Rb1, Rc,
and Rg5 attenuated neuronal apoptosis induced by glutamate in the in vitro
Huntington’s disease model through inhibition of Ca2+ signaling. Ginseng
saponins exhibited synergistic protective effects against neuronal disorders via
modulation of various neuronal signal pathways by blocking Ca2+ influx
increase (Kim et al., 2008).
Ip et al. (2015) studied anemoside A3 isolated from Pulsatilla chinensis.
Behavioral studies indicated that it not only facilitates hippocampal long-term
potentiation but also enhances spatial reference memory formation in mice,
modulating synaptic connectivity in circuits central to memory enhancement.
In summary, compounds that have the ability to both strengthen synaptic
function and facilitate neuroprotection are valuable cognitive enhancers that
may improve health and quality of life, as well as retard age-related cognitive
deterioration (Ip et al., 2015).
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Chapter 2
MINI-REVIEW:
GINSENOSIDE RG3 MODULATING
LIPID METABOLISM
Yong-Seob Jeong1 and Jisun Oh2,

1
Department of Food Science and Technology,
Chonbuk National University, Jeonju, Republic of Korea
2
School of Food Science and Biotechnology (BK21 Plus),
Kyungpook National University, Daegu, Republic of Korea
ABSTRACT
Obesity predisposes individuals to many metabolic disorders such as
diabetes, cardiovascular diseases, and various types of cancer; it shortens
life expectancy and causes economic losses. Intensive studies have been
conducted to investigate the beneficial effects of numerous food materials
which potentially control obesity. Such foods include ginseng, which has
shown promising results. Ginsenosides are the active biological
compounds in ginseng, and this review focuses on the anti-obesity effect
of ginsenoside Rg3, which, it is claimed, is responsible for the beneficial
properties of ginseng.

Corresponding Author: Jisun Oh, Ph.D. Research Professor. School of Food Science and
Biotechnology, Kyungpook National University. Daegu 41566, Republic of Korea. Phone:
+82-53-950-5752; Fax: +82-53-950-6750; Email: j.oh@knu.ac.kr.
38 Yong-Seob Jeong and Jisun Oh
Keywords: ginseng, ginsenoside Rg3, anti-obesity, adipogenesis, lipid

accumulation
1. INTRODUCTION
Ginseng, a deciduous perennial plant belonging to the family Araliaceae,
is a well-known medicinal herb. Ginseng is consumed globally as a
nutraceutical or a functional food because it has beneficial effects against
various diseases such as cancer (Helms, 2004), diabetes (Yuan, Kim, Kim and
Chung, 2012), immune disorders (Block & Mead, 2003), and
neurodegeneration (H. J. Kim, Kim and Shin, 2013). Ginseng has different
names according to its preparation: white ginseng is naturally dried; and red
ginseng is prepared by steaming followed by drying, through which process its
efficacy, preservation, and safety are enhanced (Baek, Bae and Park, 2012). In
terms of chemical composition, ginseng contains triterpene glycosides
(saponins), sesquiterpenes, acetylenic compounds (polyacetylenes), phenolic
compounds, polysaccharides, peptidoglycans, fatty acids, vitamins, etc.
(Angelova et al., 2008; Christensen, 2009). The pharmacological effects of
ginseng are primarily attributed to saponins known as ginsenosides.
Obesity is a pathophysiological condition involving excessive body fat
accumulated in adipose tissue, which increases body weight and causes
adverse health consequences with complications (Bastard et al., 2006; Cao,
2011; Spiegelman & Flier, 2001; Williams, 2013; Zamboni et al., 2005). It is a
critical risk factor that causes metabolic disorders such as arteriosclerosis,
hypertension, hyperlipidemia, and non-insulin-dependent diabetes mellitus (M.
Kim et al., 2009). Globally, an alarming increase in the prevalence of obesity
has been reported, even in children. Extensive research is being carried out to
find solutions to obesity because it causes such damage to health and the
economy. There are now several approaches to the treatment of obesity
including surgery, drugs, exercise, and diet (Laddu, Dow, Hingle, Thomson
and Going, 2011). Several studies have demonstrated that certain natural
compounds or substances found in plants, such as caffeine in oolong tea,
saponin in the roots of broad bellflower, and capsiate in sweet pepper, can be
used to treat obesity (J. H. Kim, Kang, Han and Shim, 2009).
Ginseng is known to ameliorate chronic metabolic disorders. It has been
reported that Korean red ginseng may have anti-obesity or weight-reducing
effects in obese rats (Abo-Raya, Alfky and Elgazar, 2013; J. H. Kim et al.,
2005; H. Lee, Park and Yoon, 2013; Shalaby & Hammouda, 2013; Y. B. Song
Mini-Review: Ginsenoside Rg3 Modulating Lipid Metabolism 39
et al., 2012) and humans (M. Y. Song, Kim and Kim, 2014). One study
showed that the blood lipid levels in humans taking Korean ginseng extract on
a daily basis were reduced after a couple of months (S. H. Kim & Park, 2003).
In this review, we discuss the influence on obesity of ginsenosides and their
derivatives, and focus particularly on the association between ginsenoside Rg3
and lipid metabolism.
2. PATHOPHYSIOLOGY OF OBESITY
A chronic imbalance between energy intake and energy expenditure leads
to storage of excess energy as fat in adipose tissue, which leads to obesity
(Chugh & Sharma, 2012; Spiegelman & Flier, 2001). Energy balance and
storage are homeostatically regulated by the coordination of genetic,
molecular, environmental, and psychosocial factors (Barness, Opitz and
Gilbert-Barness, 2007; Barsh, Farooqi and O'Rahilly, 2000; Spiegelman &
Flier, 2001). In a coherent physiological system, energy balance is controlled
primarily by the central nervous system, which regulates physical activity
(food intake and exercise), neuronal activity through autonomic nerves
(nutrition absorption and metabolism), and endocrine system activity
(hormones) (Barness et al., 2007; Skolnik & Ryan, 2014; Spiegelman & Flier,
2001).
Obesity is characterized by the increase of adipose tissue mass resulting
from an increased number of adipocytes (hyperplasia) and/or volume
(hypertrophy). Adipose tissue, a type of connective tissue, can be classified
into two kinds: white and brown. White adipose tissue comprises mature
adipocytes where lipid droplets (composed of triglycerides and cholesterol) are
stored, whereas brown adipose tissue is involved in thermogenesis at
specialized locations in certain animals (Bays et al., 2013).
3. REGULATION OF LIPID METABOLISM

Adipocytes are generated from preadipocytes through a differentiation
process called adipogenesis. The process is orchestrated by transcription
factors and intracellular signaling molecules in combination with extracellular
regulatory factors (Gregoire, Smas and Sul, 1998; Rosen, Walkey, Puigserver
and Spiegelman, 2000). Peroxisome proliferator-activated receptor γ (PPARγ;
known as the master adipogenic factor) and CCAAT/enhancer-binding protein

(C/EBP) family members harmoniously function in adipocyte differentiation
and intracellular lipid accumulation by modulating numerous genes related to
adipogenesis, lipid uptake, and lipid metabolism (Eeckhoute, Oger, Staels and
Lefebvre, 2012; Harms & Seale, 2013) (Figure 1).
Figure 1. Overview of regulatory pathways for anti-obesity effect of ginsenoside Rg3.
3.1. Intracellular Factors
During adipocyte differentiation, several key molecules, such as

transforming growth factor β (TGF-β) superfamily members and Wingless/int
proteins (Wnts), are involved in the intracellular signaling pathways (Huang et
al., 2011; McGregor & Choi, 2011). TGF-β stimulates the phosphorylation of
Sma- and Mad-related (Smad) proteins, which causes heterodimerization and
then translocalization of the Smad proteins into the nucleus where the Smad
complex binds to DNA in combination with other transcription factors and
influences gene expression (Son, Ka, Kim and Kim, 2014). During
adipogenesis, Smad3 phosphorylated by TGF-β binds to and inhibits C/EBPs,

which transactivate PPARγ (Choy & Derynck, 2003). Recent in vitro
(Tsurutani et al., 2011; Zamani & Brown, 2011) and in vivo (Clouthier,
Comerford and Hammer, 1997) studies have consistently shown that TGF-β
signaling leads to inhibition of the early stage of preadipocyte differentiation
and lipid accumulation.
In addition, the Wnt/β-catenin pathway, which generally controls cell fate,
is activated in preadipocytes and Wnts are expressed at relatively low levels
following induction of differentiation (Bennett et al., 2002; Bowers & Lane,
2008). It has been suggested that Wnt signaling may regulate the late stage of
adipogenesis through the lymphoid-enhancer-binding factor/T-cell-specific
transcription factor (LEF/TCF) family of transcription factors that modulate
Wnt target gene expression (Singh et al., 2006).
3.2. Extracellular Factors
In terms of hormonal regulation, leptin, ghrelin, insulin, sex hormones,

and growth hormone influence the appetite, energy balance, and body fat
distribution. Disturbances in the balance of these hormones contribute to
obesity. Insulin predominantly interacts with insulin growth factor-1 (IGF-1)
receptor and consecutively activates downstream signaling molecules
including insulin receptor substrate (IRS), phosphoinositide 3-kinase (PI3K),
and AKT/protein kinase B (PKB), which eventually activates C/EBPα (Son et
al., 2014). Another way for insulin to deliver its signal for adipogenesis is
through mitogen-activated protein kinase (MAPK) pathways that activate
extracellular signal-regulated kinases (ERKs) and p38 (Bost, Aouadi, Caron
and Binetruy, 2005), modulating the activity of PPARα or PPARγ (Son et al.,
2014). Alternatively, IRS signaling promotes cAMP response element-binding
(CREB) phosphorylation (Tseng et al., 2005), or AKT/PKB signaling inhibits
the anti-adipogenic transcription factor, forkhead box protein O (FOXO)
(Nakae et al., 2003). Furthermore, adipocytes themselves secrete various
hormones, growth factors, and cytokines that act as signals for adipocyte
metabolism (Fruhbeck, Gomez-Ambrosi, Muruzabal and Burrell, 2001).
3.3. AMP-Activated Protein Kinase in Lipid Metabolism
AMP-activated protein kinase (AMPK), a serine/threonine protein kinase,

is also known to be a key molecule in energy metabolism (Daval, Foufelle and
Ferre, 2006). Food intake is regulated according to AMPK activity in the
hypothalamus; low energy levels resulting from fasting or exercise (low leptin,
high ghrelin) activate AMPK leading to increased appetite, whereas high
energy levels resulting from eating (high leptin, low ghrelin) suppress AMPK
activity, thereby reducing appetite (Daval et al., 2006; Kola, Boscaro, Rutter,
Grossman and Korbonits, 2006).
In the metabolic organs, such as muscle and liver, AMPK plays a role as a
cellular energy gauge in controlling energy homeostasis by switching on ATP-
generating catabolic pathways (glucose uptake, fatty acid oxidation) and
switching off ATP-consuming anabolic pathways (gluconeogenesis,
lipogenesis) (Hue & Rider, 2007; Kahn, Alquier, Carling and Hardie, 2005;
Yin, Mu and Birnbaum, 2003). During lipid metabolism, AMPK is activated
by adipocyte-secreted leptin or adiponectin under lipolytic conditions. The
activated AMPK inhibits the synthesis and degradation of the lipids in
adipocytes, resulting in reduced availability of free fatty acids (Kahn et al.,
2005). In the cholesterol biosynthesis pathway in particular, AMPK
phosphorylates and inactivates 3-hydroxy-3-methylglutaryl-CoA reductase
(HMGCR), a key enzyme for cholesterol synthesis.
4. GINSENOSIDES
Ginsenosides, which are triterpene saponins, are the biologically active
components of ginseng; the beneficial effects of ginseng are attributable to the
function of ginsenosides (Attele, Wu and Yuan, 1999; Qi, Wang and Yuan,
2010; Tawab, Bahr, Karas, Wurglics and Schubert-Zsilavecz, 2003).
Approximately 150 different ginsenosides are classified into either of two
groups, dammarane or oleanane, depending on the aglycone to which the
water-soluble sugar moieties are attached (Qi, Wang and Yuan, 2011). Based
on their chemical structure, the dammarane ginsenosides fall into one of two
types: protopanaxadiol (PPD), including Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc,
Rd, Rg3 and Rh2; or protopanaxatriol (PPT), including Re, Rg1, Rg2, Rf, and
Rh1 (Jia, Zhao and Liang, 2009).
Pharmacopoeias from various countries include information on natural or
processed ginseng and indicate the required proportions of ginsenosides Rg1,
Rb1, and/or Rg3. For example, the USA requires > 0.2% of Rg1 and > 0.1%
Rb1; Europe requires > 0.4% of Rg1+ Rb1; China requires > 0.3% of Rg1+Re
and > 0.2% of Rb1; and Korea and Japan require > 0.1% of Rg1 and > 0.2% of
Rb1. The Korean health functional food code stipulates that the standard of
ginsenoside content in red ginseng should be 2.5–34 mg/g combined contents
of Rg1, Rb1, and Rg3.
4.1. Characteristics of Ginsenoside Rg3
Minor but highly absorbable and bioactive ginsenosides, such as Rg3,

Rh2, Rh1, and C-K, can be produced by hydrolysis of the major ginsenosides,
such as Rb1, Rb2, Rc, Rd, Re, and Rg1 (Ji et al., 2001). The hydrolytic
reaction is processed by heating, acid treatment, enzymatic digestion, or
microorganism-mediated conversion. In particular, several studies have
demonstrated that Rg3 can be formed by deglycosylation of Rb1, Rb2, and Rc
by heat-processing (E. H. Park et al., 2014; C. Z. Wang et al., 2008), mild acid
treatment (Han et al., 1982), or intestinal bacteria (Bae, Han, Choo, Park and
Kim, 2002), and can be further metabolized into Rh2 or PPD (Bae et al.,
2002).
Ginsenoside Rg3 exists as S or R isomers depending on the spatial
position of the hydroxyl group on the chiral carbon (C-20) of the aglycone;
20(S)-Rg3 and 20(R)-Rg3 are epimers (Cheng, Na, Bang, Kim and Yang,
2008). The epimers have stereospecific properties that result in different
biological activities. The structure of 20(S)-Rg3 is more compact than that of
20(R)-Rg3; the orientation of the alkene chain attached to the C-20 of 20(S)-
Rg3 is less flexible and therefore more tightly packed (Kang, Yokozawa,
Yamabe, Kim and Park, 2007). Solubility in water is thought to be influenced
by this structural difference; the hydroxyl group of the 20(S)-Rg3 isomer,
which is more stable and compact, is more accessible to water than that of the
20(R)-Rg3 isomer. The 20(S)-Rg3 isomer, but not the 20(R)-Rg3 isomer, can
inhibit voltage-gated Ca2+, K+, and Na+ channels, whereas both isomers are
able to antagonize the 5-HT3A serotonin and α3β4 nicotinic acetylcholine
receptors (Jeong et al., 2004). Compared with 20(S)-Rg3, 20(R)-Rg3 is known
to have higher antioxidative activity in mice subjected to cyclophosphamide
injection-induced oxidative stress (Wei, Su, Su, Hu and Hu, 2012). In addition,
20(R)-Rg3 demonstrates angio-suppressive activity in human umbilical vein
endothelial cells (Yue et al., 2006), and suppresses TGF-β1-induced cell
migration, invasion, and anoikis resistance when tested in A549 human lung
cancer cells by inhibiting the epithelial–mesenchymal transition process (Y. J.
Kim et al., 2014).
4.2. Anti-Obesity Effect of Ginsenoside Rg3
Accumulation of triglycerides and cholesterol correlates to various

metabolic disorders and cardiovascular diseases leading to a reduction of life
expectancy (Celermajer, Chow, Marijon, Anstey and Woo, 2012; Van Gaal,
Mertens and De Block, 2006). Ginsenoside Rg3 is known to inhibit lipid
accumulation by reducing adipogenesis via AMPK activation (Figure 1). Lee
et al. (2011) showed that Rg3 increased glucose uptake in 3T3-L1 cells,
possibly through GLUT4, in which expression was concurrently increased (O.
H. Lee, Lee, Kim and Lee, 2011). Lee et al. (2012) reported that Rg3 activated
AMPK and thus reduced triglyceride and cholesterol levels in HepG2 cells (S.
Lee, Lee, Kim, Kim and Kim, 2012). Hwang et al. (2009) showed that
adipocyte differentiation of 3T3-L1 cells was inhibited by Rg3 treatment in a
dose-dependent manner through activation of AMPK as well as inhibition of
PPARγ activity (Hwang et al., 2009). Hwang et al. (2007) reported that
ginsenoside Rh2, formed from Rg3 by deglycosylation at the C-3 position,
effectively reduced cell viability and adipocyte differentiation of preadipocytes
by inhibiting PPARγ and activating AMPK (Hwang et al., 2007). Thus, these
findings demonstrate that Rg3 and its derivative Rh2 reduce obesity by
influencing AMPK activity and/or PPARγ signaling pathways.
Interestingly, Rb1, from which Rg3 and Rh2 are derived, was also shown
to inhibit triglyceride accumulation in 3T3-L1 adipocytes by the stimulation of
protein kinase A (PKA)-dependent pathways (S. Park, Ahn, Kwon, Ko and
Jun, 2008). However, the authors of other studies have reported that Rb1
increases adipogenesis or inhibits lipolysis in adipocytes by antagonizing
PPARγ and C/EBPα gene expression (Shang et al., 2007; H. Wang, Reaves
and Edens, 2006). These discrepancies among the studies may be attributed to
the different concentrations of the ginsenosides tested, suggesting the necessity
for a systematic investigation on the function of each ginsenoside at the
molecular and cellular levels. Therefore, the conversion of Rb1 in certain food
materials to Rg3 or further to Rh2 is critical for obtaining functional,
bioavailable substances that are capable of antagonizing obesity-related
singling pathways, thereby inhibiting lipid accumulation in adipocytes.
CONCLUSION
Obesity is a high risk factor for many diseases including type 2 diabetes,
cardiovascular disorders, and even cancer. Numerous studies imply that
various types of ginseng extracts are beneficial for reducing obesity. Many
investigators have made an enormous effort to discover how ginseng actually
works in the human body. It is now known which regulatory mechanisms in
adipocytes function in response to obesity-related hormones, and which
ginsenosides are mostly related to the modulation of lipid metabolism.
Ginsenoside Rg3 is an abundant and bioavailable constituent of natural
and processed ginseng. Rg3 is known to be effective in several disorders,
including cancer and obesity. Thus, enhancing the Rg3 component of natural
resources containing ginsenosides would be an important research topic for the
development of functional food materials. Until now, most studies on Rg3
have focused on transcription-regulating factors. Further studies on
orchestrating the downstream regulatory molecules that are stimulated by Rg3
may provide insight into the systematic mechanism associated with obesity,
and may lead to the improvement of obesity treatment or prevention.
ACKNOWLEDGMENTS
This work was supported by a Basic Science Research Program through
the National Research Foundation of Korea funded by the Ministry of
Education (2013R1A1A2013362).
Author Disclosure Statement
The authors declare no conflict of interest.
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BIOGRAPHICAL SKETCH
Jisun Oh, PhD

Majors in Neuroscience and Cell Biology
E-mail: j.oh@knu.ac.kr; ohjisunoh@gmail.com
Educational Experience
2011.04 – Post-Doctoral Fellow at Molecular Neurobiology Lab,

2013.03 McLean Hospital / Harvard Medical School, Belmont, MA,
The United States
Principal Investigator: Kwang-Soo Kim, Ph.D.
2004.08 – Ph.D. in Neuroscience and Biomedical Sciences
2010.12 (Specialty in Cell Biology),
Iowa State University, Ames, IA, The United States
Supervisor: Donald S. Sakaguchi, Ph.D.
Thesis Title: ‘Astrocyte-derived soluble factors promoting
neuronal differentiation of adult neural progenitor cells’
2001.03 – M.S. in Animal Science and Biotechnology
2003.02 (Specialty in Molecular Genetics),
Kyungpook National University, Daegu, South Korea
Supervisor: Jinkyu Lim, Ph.D.
Thesis Title: ‘A Functional Study of Calretinin in Rat Brain’
1996.03 – B.A. in Animal Science and Biotechnology,
2001.02 Kyungpook National University, Daegu, South Korea
Professional Experience
Research Experience
2015.03 – Research Professor, School of Food Science and

Present Biotechnology, Kyungpook National University,
Daegu, South Korea
- Studying cellular properties of induced cancer stem
cells from various cancer cells
- Studying various functions of soybean-derived
flavonoids in alleviating cancer development
- Studying neuroprotective effect of ginsenosides
from hydrolyzed Korean red ginseng
2013.04 – Research Professor, Research Center for Industrial
2015.02 Development of Biofood Materials, Chonbuk National
University,
Jeonju, Jeollabuk-do, South Korea
- Studying cellular properties of induced cancer stem
cells from various cancer cells
- Studying anti-cancerous effect of ginsenosides from
fermented Korean red ginseng
- Studying hippocampal neural cell dysfunction in
association with cytosolic inclusion of TDP-43
proteins
2011.04 – Postdoc Fellow, Molecular Neurobiology Lab, McLean
2013.03 Hospital / Harvard Medical School, Belmont, MA, USA
- Characterizing cellular and phenotypic properties of
induced pluripotent stem cells (iPSCs) derived from
psychiatric disorder subjects as the iPSCs give rise to
specific neural lineages
2005.07 – Research Assistant, Dept. of Genetics, Development
2010.08 and Cell Biology, Iowa State University
- Studying functional properties of the cellular
membrane of neural progenitors differentiated into
neurons using an electrophysiological approach,
whole cell patch clamping
- Characterizing cellular properties of adult rat
hippocampal neural progenitors formed in
neurospheres in vitro and in vivo after transplantation
into vitreous body of neonatal rat eyes
2004.08 – Research Assistant, Dept. of Biomedical Sciences, Iowa

2005.06 State University
- Involved in studying neuronal circuitry in an
epithalamic area of the rat brain using whole cell
patch clamp analysis
2003.03 – Research Intern, Korea Science and Engineering Foundation
2004.02 - Mapping and standardizing the proteome from
human urine using proteomic approaches
2001.03 – Research Assistant, Lab. of Molecular Genetics, Kyungpook
2003.02 National University
- Characterizing the function of a calcium-binding
protein, calretinin, highly expressed in the superior
colliculus of the rat brain, using proteomic and
molecular genetic approaches
- Involved in searching for the proteins related to
circadian rhythm and sleep-wake cycle in the rat brain
2000.07 – Summer Undergraduate Research Fellow, Lab. of Molecular
2000.08 Neurophysiology (under supervision by Dr. Kyong-Tai
Kim), Pohang University of Science and Technology
- Involved in studying the role of serotonin N-
acetyltransferase in depression
1999.07 – Undergrad research volunteer, Lab. of Molecular Genetics,
2001.02 Kyungpook National University
- Mapping rat brain proteome using two-dimensional
sodium dodecyl sulfate(SDS)-polyacrylamide gel
electrophoresis(PAGE) and mass spectrometric
analysis
- Involved in characterizing a bovine pregnant-
associated protein from bovine urine and in
developing diagnostic agent to detect bovine
pregnancy
Teaching Experience
2016.03 – Adjunct Instructor (Theory and Practice of Food

2016.06 Biomaterials), School of Food Science and
Biotechnology, Kyungpook National University
2015.09 – Adjunct Instructor (Advanced Food Biotechnology; Food
2015.12 Biomaterials), School of Food Science and
Biotechnology, Kyungpook National University
2015.03 – Adjunct Instructor (Food Material Analysis Lab), School

2015.06 of Food Science and Biotechnology, Kyungpook National
University
2007.08 – Graduate Mentor for honors students, Iowa State
2010.12 University
2006.08 – Teaching Assistant (Human Anatomy and Physiology),
2010.12 Dept. of Genetics, Development and Cell Biology, Iowa
State University
Main Instructor: Barbara Krumhardt, Ph.D.
Awards and Honors
2004.08 – Research/Teaching Assistantship

2010.12 from Iowa State University
2010 Nominee for Graduate Student Travel Awards
from Ames, Iowa Chapter of the SfN
2007, 2009, Travel Awards
2010 from Dept. of Genetics, Development and Cell Biology,
Iowa State University
2001 – 2003 Research Assistantship
from Kyungpook National University
2002 Scholarship for highest honor graduate students
from Kyungpook National University
1996 – 1998, Scholarship for highest honor undergraduate students
2000 from Kyungpook National University
Professional Activities
2014 – Present Member, The Society for Neuroscience

2013 – Present Member, The Korean Society of Food Science and
Nutrition
2006 – 2010 Student member, The Society for Neuroscience
Publications
In preparation
1. Jo GR†, Oh J†, Kwon CS, and Kim JS, (2016) Activation of Nrf2
Signaling Pathway by Low Dose of Glyceollins is Mediated by p53 in
Human Colon Cancer Model. J Agric Food Chem (Under review)
2. Lee HY, Park YM, Kim J, Oh HK, Choi NY, Kim KS, Kang HJ, Kim
RR, Yang HJ, and Oh J*. (2016) Orostachys japonicus A. Berger
extracts induce immunity-enhancing effects on cyclophosphamide-
treated immunosuppressed rats. J Ethnopharmacol (Under review)
3. Lee HY, Oh HG, Kang YG, Park YM, Kim YP, Moon DI, Kim YB,
Hong S, Jeong YS, Oh J* and Kim O*. (2016) Protective effect of
epidermal growth factor on 5-fluorouracil-induced small intestinal
damage in vivo. Histol Histopathol (Under review)
4. Shin GY, Oh JG, Woo Y, Lee H, Jeong YS, Kim JS, and Oh J*.
(2016) Antioxidant potential of selected edible plants in Korea. Appl
Biol Chem (Under review)
5. Kim D, Hlakty L, Jeong YS and Oh J*. (2016) Mini-review: Impact
of anti-cancer phytochemicals on cancer stem cells. MDPI-Toxins
(Under revision)
Published in 2014 – Present
1. Jang JH, Seo JY, Oh J, Kim JS, Kim EJ, and Kim JS. (2016) In vitro
and in Vivo anti-inflammatory activity of mixed fruit and vegetable
juice. Food Sci Biotechnol, 25:1-5.
2. Seo JY, Ju SH, Oh J, Lee SK, Kim JS. (2016) Neuroprotective and
Cognition Enhancing Effects of Compound K Isolated from Red
Ginseng. J Agric Food Chem doi: 10.1021/acs.jafc.5b05789
3. Yum EJ, Lee NK, Oh J, Lee CH, Oh MJ, Kim HR, Oh CH, Park JH,
Moon HJ and Jeong YS. (2016) Anti-Obesity Effect of a Mixture of
Skim Milk, Red Ginseng Extract, and Black Raspberry Extract
Fermented with Lactobacillus acidophilus on High-Fat Diet-Fed
Obese Mice. J Food Chem Nanotechnol, 2: 65-71.
4. Lee NK, Jeong JH, Oh J, Kim Y, Ha YS and Jeong YS. (2015)
Conversion of flavonols kaempferol and quercetin in mulberry
(Morus alba L.) leaf using plant-fermenting Lactobacillus plantarum.
J Food Biochem, 39: 765-770.
5. Lee Y, Oh J*, Lee H, Lee NK, Jeong DY and Jeong YS*. (2015)
Lactic acid bacteria-mediated fermentation of Cudrania tricuspidata
leaf extract improves its antioxidative activity, osteogenic effects, and
anti-adipogenic effects. Biotechnol Bioproc Eng, 20:861-870.
6. Lee Y, Oh J* and Jeong YS. (2015) Lactobacillus plantarum-mediated
conversion of flavonoid glycosides into flavonols, quercetin and
kaempferol, in Cudrania tricuspidata leaves. Food Sci Biotechnol,
24:1817-1821.
7. Park YM, Lee BG, Park SH, Oh HG, Kang YG, Kim OJ, Kwon LS,
Kim YP, Choi MH, Jeong YS, Oh J* and Lee HY*. (2015) Prolonged
Oral Administration of Gastrodia Elata Extract Improves Spatial
Learning and Memory of Scopolamine-Treated Rats. Lab Anim Res,
31:69-77.
8. Jeong EJ, Lee NK, Yum EJ, Nam K, Oh J, Kim YS, Park JY, Kim SJ,
Jeong YS. (2015) Effect of calcium chloride on the texture of pickled
radish wrap. Kor J Food Preserv, 22:452-457.
9. Oh J, Jeon SB, Lee Y, Lee H, Kim J, Kwon BR, Yu KY, Cha JD,
Hwang SM, Choi KM and Jeong YS. (2015) Fermented red ginseng
extract inhibits cancer cell proliferation and viability. J Med Food,
18:421-428.
10. Oh J, Daniels G, Chiou LS, Ye EA, Jeong YS and Sakaguchi DS.
(2014) Multipotent adult hippocampal progenitor cells maintained as
neurospheres favor differentiation toward glial lineages. Biotechnol J,
9:921-933. (Cover image for the issue of 7)
11. Jeong EJ, Lee NK, Oh J, Jang SE, Lee JS, Bae IH, Park JH, Jung HK
and Jeong YS. (2014) Inhibitory effects of cinnamon essential oil on
selected cheese-contaminating fungi (Penicillium sp.) during cheese-
ripening process. Food Sci Biotechnol, 23:1193-1198.
Published in 2003 – 2013
1. Howk CL, Levine HA, Smiley MW, Mallapragada SK, Nilsen-

Hamilton M, Oh J and Sakaguchi DS. (2012) A mathematical model
for selective differentiation of neural progenitor cells on
micropatterned polymer substrates. Math Biosci, 238:65-79.
2. Petersen LK, Oh J, Sakaguchi DS, Mallapragada SK and Narasimhan
B. (2011) Amphiphilic Polyanhydride Films Promote Neural Stem
Cell Adhesion and Differentiation. Tissue Eng Part A, 17:2533-2541.
3. Ariza CA, Fleury AT, Tormos CJ, Petruk V, Chawla S, Oh J,

Sakaguchi DS and Mallapragada SK. (2010) The influence of electric
fields on hippocampal neural progenitor cells. Stem Cell Rev, 6:585-
600.
4. Oh J, McCloskey MA, Blong CC, Bendickson L, Nilsen-Hamilton M
and Sakaguchi DS. (2010) Astrocyte-derived interleukin-6 promotes
specific neuronal differentiation of neural progenitor cells from adult
hippocampus. J Neurosci Res, 88:2798-2809.
5. Perdian DC, Cha S, Oh J, Sakaguchi DS, Yeung ES and Lee YJ.
(2010) In situ probing of Cholesterol in Astrocytes at the single cell
level using Laser Desorption Ionization Mass Spectrometric imaging
with Colloidal Silver. Rapid Commun Mass Spectrom, 24:1147-1154.
6. Blong CC, Jeon CJ, Yeo JY, Ye EA, Oh J, Callahan JM, Law WD,
Mallapragada SK and Sakaguchi DS. (2010) Differentiation and
Behavior of Human Neural Progenitors on Micropatterned Substrates
and in the Developing Retina. J Neurosci Res, 88:1445-1456.
7. Oh J, Recknor JB, Recknor JC, Mallapragada SK and Sakaguchi DS.
(2009) Soluble factors from neocortical astrocytes enhance neuronal
differentiation of neural progenitor cells from adult rat hippocampus
on micropatterned polymer substrates. J Biomed Mater Res A,
91:575-585.
8. Oh J, Pyo JH, Jo EH, Hwang SI, Kang SC, Jung JH, Park EK, Kim
SY, Choi JY and Lim J. (2004) Establishment of a near-standard two-
dimensional human urine proteomic map. Proteomics, 4:3485-3497.
9. Choi BH, Chae HD, Park TJ, Oh J, Lim J, Kang SS, Ha H and Kim
KT. (2004) Protein kinase C regulates the activity and stability of
serotonin N-acetyltransferase. J Neurochem, 90:442-454.
10. Pyo JH, Hwang SI, Oh J, Lee SJ, Kang SC, Kim JS and Lim J. (2003)
Characterization of a bovine pregnancy-associated protein using two-
dimensional gel electrophoresis, N-terminal sequencing and mass
spectrometry. Proteomics, 3:2420–2427.
11. Oh J and Lim J. (2003) Improvement of in-gel digestion and recovery
of peptides for peptide mass fingerprinting. Agric Chem Biotechnol,
46:1-5.
Presentations (Selected)
1. Kim M, Kang S, Jo G, Jang J, Jeong G, Ju S, Seo H, Woo Y, Oh J,

Kim JS. Methylene Chloride Fraction of Salicornia herbacea L.
Suppresses Glutamate-induced Neuronal Cytotoxicity and Improves

Galactose-induced Learning and Memory Impairment. Experimental
Biology, Apr 2016.
2. Jo G, Oh J, Kang S, Kim M, Jang J, Jeong G, Seo H, Ju S, Woo Y,
Lim S, Kim JS. Soybean-derived glyceollins promote Nrf2mediated
detoxifying enzymes in colorectal cells. Experimental Biology, Apr
2016.
3. Lee H, Jeong JH, Lee NK, Jeong YS, Oh J. Camellia japonica extract
suppresses cancer stem cell survival and proliferation. Int. Conf. on
Food Factors (ICoFF), Nov 2015.
4. Oh J, Kim JS, McCloskey MA, and Sakaguchi DS. Effects of calcium
influx on interleukin-6-mediated neuronal differentiation of neural
progenitor cells. SfN Ann Mtg, Oct 2015. (session # 475.05)
5. Seo HL, Seo JY, Kang SJ, Jo GR, Ju SH, Jeong GI, Oh J and Kim JS.
Soybean-derived Glyceollins Inhibit TNFα-mediated Colon Cancer
Cell Proliferation. Kor Soc of Food Sci Technol, Jun 2015.
6. Jo GR, Seo JY, Kang SJ, Jang JH, Jeong GI, Seo HL, Ju SH, Oh J and
Kim JS. Anti-cancer Effect of Soybean-derived Glyceollins by
Induction of Phase II Detoxifying Enzymes in Colorectal Cancer
Cells. Kor Soc of Food Sci Technol, Jun 2015.
7. Oh J, Lee Y, Lee H, Kim J, Kwon BR, Yu KY, Cha JD, Hwang SM,
Choi KM and Jeong YS. Anti-cancer effect of red ginseng extract
fermented by Lactobacillus rhamnosus. Kor Soc for Food Sci Nutr,
Oct 2014.
8. Oh J, Daniels G, Chiou LS, Ye EA, Jeong YS and Sakaguchi DS.
Characterization of Multipotent Adult Rat Hippocampal Progenitor
Cells Maintained as Neurospheres: Differentiation in vitro and in vivo
Following Transplantation. Kor Soc for Biotechnol Bioeng, Apr 2014.
9. Lee Y, Oh J, Jeon SB, Jeong EJ, Lee H, Kim M and Jeong YS.
Antioxidant activity and osteogenic effects of Cudrania tricuspidata
hot water extract fermented by Lactobacillus planetarium SLD 1413.
Kor Soc for Biotechnol Bioeng, Apr 2014.
10. Oh J, Jeon SB, Lee Y, Lee H, Kim J, Kwon BR, Yu KY, Cha JD,
Hwang SM, Choi KM and Jeong YS. Inhibitory effect of fermented
red ginseng extract on cancer cell proliferation and viability. Kor Soc
for Biotechnol Bioeng, Apr 2014.
11. Moon JI, Oh J, Chung S, Naujock M, Mattis VB, Svendsen CN,
Arjomand J, Kim KS. Efficient and fast in vitro differentiation of
human pluripotent stem cells (hPSCs) into different neural and glial
linages using EZ sphere-based methods. Soc for Neurosci (SfN) Ann

Mtg, Oct 2012. (session # 633.03)
12. Oh J, Daniels GJ, Chiou LS, Ye EA, and Sakaguchi DS.
Characterization of multipotent adult rat hippocampal progenitor cells
maintained as neurospheres: Differentiation in vitro and in vivo
following transplantation. SfN Ann Mtg, Nov 2010. (session # 839)
13. Oh J, McCloskey MA, and Sakaguchi DS. Astrocyte-derived
interleukin-6 promotes specific neuronal differentiation of neural
progenitor cells from adult hippocampus. SfN Ann Mtg, Oct 2009.
(session # 507.19)
14. Oh J, McCloskey MA, Blong CC, and Sakaguchi DS. Astrocyte-
derived interleukin-6 promotes specific neuronal differentiation of
neural progenitor cells from adult hippocampus. Midwest
Neurobiology Mtg (SfN-Milwaukee Chapter), Apr 2009.
15. Oh J, McCloskey MA, and Sakaguchi DS. Astrocyte-derived soluble
factors induce specific neuronal differentiation of neural progenitor
cells from adult hippocampus. SfN Ann Mtg, Nov 2007. (session #
673.6)
16. Blong CC, Oh J, Kim HH, Jeon CJ, Ceraci C, Nilsen-Hamilton M,
and Sakaguchi DS. Stimulation of specific neuronal differentiation of
adult hippocampal progenitor cells by astrocyte-derived IL-6. SfN
Ann Mtg, Nov 2007. (session # 673.7)
17. Ye EA, Blong CC, Oh J, Jeon CJ, Mallapragada SK, and Sakaguchi
DS. Growth and differentiation of human neural progenitor cells on
micropatterned polymer substrates. SfN Ann Mtg, Nov 2006.
Research Support
Ongoing
Basic Science Research Program-Research Fellow Project, Ministry of

Education
(Project # 2013R1A1A2013362)
2013.06 – 2016.05
Role: PI
Title: Screening of plant extract-derived effective components as potent
anticancer agents using induced cancer stem cells
Completed
Technological Innovative Project, Ministry of Trade, Industry, and Energy

(Project # 10045726)
2013.05 – 2014.04
Role: PI for unit research project
Title: Development of convergence products using by Jeonbuk regional
resources
Chapter 3
APPLICATION OF SAPONINS
IN REMEDIATION OF METAL-
CONTAMINATED SOILS
Zygmunt Mariusz Gusiatin*

Faculty of Environmental Sciences, Department of Environmental
Biotechnology, University of Warmia and Mazury in Olsztyn,
Olsztyn, Poland
ABSTRACT
Saponins represent glycosides, secondary plant metabolites. Their
diverse physicochemical properties have been successfully used in a
number of commercial applications in the food, cosmetics, agricultural
and pharmaceutical sectors. Recently, saponins have become more and
more attractive in environmental applications. Due to their amphiphilic
nature and natural origin, they can behave as biosurfactants. Saponins are
able to foam and to effectively decrease surface/interfacial tension. These
properties can be used in remediation projects to remove organic and
inorganic pollutants from soil.
The present chapter presents chemical structure of saponins, their
main sources and surface active properties, including foam production
*
Corresponding Author: Zygmunt Mariusz Gusiatin. Faculty of Environmental Sciences,
Department of Environmental Biotechnology, University of Warmia and Mazury in
Olsztyn, Sloneczna 45G, 10-719 Olsztyn, Poland. Email: mariusz.gusiatin@uwm.edu.pl.
64 Zygmunt Mariusz Gusiatin
and its stability, wetting ability, surface tension and critical micelle
concentration (CMC). Based on recent literature, examples of using
saponins from different sources to remove metals from soils, sludges and
fly ashes are presented. For metal removal, saponins can be used in form
of solution, foam or microbubbles. The advantage of using saponins in
soil remediation is also their ability to simultaneously remove combined
pollutants, e.g., heavy metals and polichlorinated biphenyls (PCBs) or
polycyclic aromatic hydrocarbons (PAHs). A promising alternative for
remediation of these pollutants can be a mixing of saponin with chelating
agents. Despite the high efficiency of saponins to remove pollutants from
soil, they are still expensive agents. This chapter demonstrates methods of
saponins recovery after soil washing and attempts of their reuse. A new
trends of using saponins in soil remediation are also presented.
Keywords: heavy metals, soil remediation, saponins
1. CHEMICAL STRUCTURE OF SAPONINS

AND THEIR SOURCES
Saponins are secondary plant metabolites with a complex and

heterogenous structure. They are amphiphilic, comprising one or more
hydrophobic aglycones (also called sapogenins) and one or more hydrophilic
glycones (Zhou et al. 2013). Usually, saponins occur as a mixture of
glycosides with either one or several aglycones, so it is difficult to determine
their exact chemical structure. Aglycons contain –OH, –COOH and –CH3
functional groups, and glycones contain one or more sugar chains, in which
glucose, galactose, glucuronic acid, rhamnose, arabinose, xylose, apiose or
fucose can be found (Oleszek and Hamed 2010, Güçlü-Üstündag and Mazza
2007).
Saponins are classified into two basic groups, triterpene and steroidal,
based on the chemical structure of the aglycone (Figure 1). Depending on the
number of sugar chains in the glycone, saponins are also classified as mono-,
di- and tridesmosidic. Monodesmosidic saponins have a single sugar chain,
normally attached at C-3. Bidesmosidic saponins have two sugar chains, often
with one attached by an ether linkage at C-3, and one by an ester linkage at C-
28 (triterpene saponins) or by an ether linkage at C-26.
Application of Saponins in Remediation of Metal-Contaminated Soils 65
30
21
22
11 O
28
16
O
24 23 HO
a b
Figure 1. Structure of saponin aglycones: a) triterpene, b) steroidal (Güçlü-Üstündag

and Mazza 2007).
Saponins are widely distributed in plants belonging to nearly 100 families,

in various geographical and climatic zones around the world, from polar
tundra to tropical forests and wastelands (Szakiel et al. 2011). They can be
found in plant roots, shoots, flowers and seeds. Saponins also occur in some
algae and lower marine organisms (Oleszek and Hamed 2010). Although
saponins are present in both edible and inedible plants, their amount differs in
various sources. Among the edible plants, saponins are found in soya bean,
chickpea, mungbean, peanuts, lentils, beans, oats, leek, garlic, asparagus, tea,
spinach, sugar beet, sesame and yam, among others (Oleszek and Hamed
2010). As an example, alfalfa (Medicago sativa L.) contains triterpene
saponins (Pecetti et al. 2006), but their content is below 2% (Fenwick et al.
1991). Soybeans (Glycine max L.) have a saponin content from 0.2 to 0.5%
(Güçlü-Üstündag and Mazza 2007), or even as high as 6.2% (Shiraiwa et al.
1991). The seed coats and seed hulls of quinoa (Chenopodium quinoa Willd.)
contain between 0.15 and 2.3% saponin (Güçlü-Üstündag and Mazza 2007).
Sugar beet (Beta vulgaris L.) also contains up to 0.6% triterpene saponins.
As can be seen above, the content of saponins in edible plants is not high.
Therefore, saponins from nonfood sources are used for commercial purposes.
Certain plant species are rich in saponins. Madhuca longifolia is a large
evergreen tree extensively cultivated for its oil-containing seeds in warm
climates from India to New Guinea. The seeds contain triterpene saponins,
with a concentration of around 10% (Scientific Opinion… 2009). After oil
extraction from the seeds, the press cake can contain over two times more
saponins than the seeds (Scientific Opinion… 2009). Another source of
saponins is Yucca schidigera. This plant is cultivated in the southwestern
United States and Mexico and is a material for soap production (Oleszek and
Hamed 2010). Yucca schidigera has the highest saponin content of any of the
Yucca species. The saponin concentration in this plant is up to 10% (Oleszek
2000). An important source of saponins is Quillaia saponaria Molina, found
in the arid areas of Chile. The saponins are usually found in the bark of the
tree and their content ranges from 10% dry weight (San Martin and Briones
1999) to over 25% (Oleszek and Hamad 2010). Two different extracts of
quillaia bark, both containing triterpene saponins, were assessed by the Joint
FAO/WHO Expert Committee on Food Additives (JECFA) (Scientific
Opinion… 2009):
 “unpurified extract” of Quillaia (type 1), which is obtained by

aqueous extraction from the milled inner bark or from the wood of
pruned stems and branches of Q. saponaria Molina (family Rosaceae),
followed by clarification and purification; the product has a saponin
content of 2.0-2.6% and contains triterpenoid saponins consisting
predominantly of four different glycosides of quillaic acid;
 “semi-purified extract” of Quillaia (type 2), which is obtained by
subjecting the aqueous extract to several clarification and purification
steps (ultra-filtration or affinity chromatography), yielding a saponin
content of 7.5-8.0%).
Quillaia saponaria and Yucca schidigera extracts, containing triterpene

and steroidal saponins, respectively, are regarded as safe products and are
currently permitted even as food additives.
Tea saponin (TS) is distributed in various parts of the Camellia species.
Extract from Camellia oleifera seeds has been used as a natural detergent and
is commercially utilized as a foam-stabilizing and emulsifying agent (Chen et
al. 2010). Camellia oleifera is an important source of edible oil in tropical and
sub-tropical regions of Asia, especially in China. In China, about ten million
tons of oil-tea cake is produced per year, and at least three million tons of tea
saponin may be produced each year, which needs to be managed (Xia et al.
2009). This waste can be degraded automatically without a toxic effect on
soils. Hydrophilic groups of TS contain hydroxyl groups and ester groups
(Wang e t al. 2015).
Saponin from Sapindus mukorossi, which grows in China and Japan, is
also used as a natural detergent. This saponin is extracted from the fruit’s
pericarp. This type of saponin has been listed in the Japanese Cosmetic
Ingredient Codex and is authorized by the Ministry of Health and Welfare of
Japan for use as an ingredient in cosmetics. Saponin content in S. mukorossi

can be 20% (Oleszek and Hamed 2010). The most common structure in the
extract is an oleanane-type triterpenoid saponin. This is composed of
aglycones in the form of hederagenin, which is substituted with a di- or
trisacharide at C-3.
Another source of saponins is Saponaria officinalis, commonly known as
soapwort or bouncing bet. This an invasive perennial herb that was introduced
to the US from Europe, and is now distributed across the continental US and
northern Canada (Jabbari et al. 2013). It is found along roadsides, railroad
tracks and in waste places. Its foliage contains a glycoside for wetting,
foaming and grease dispersion (Oleszek and Hamed 2010). S. officinalis
contains saponins in both the roots and leaves. In the past, this plant was used
to manufacture soap.
Aescin is one of the most frequently encountered saponin preparations,
commercially produced from seeds of the horse chestnut tree (Aesculus
hippocastanum L.) (Hostettmann and Marston 1995, Pekdemir et al. 1999).
Horse chestnut seeds are rich in saponins (3-5%), over thirty of which have
been isolated and identified. The main compound is aescin – a mixture of
acylated triterpene glycosides. Three fractions of aescin, denoted as crypto-, α-
and β-aescin have been described in the literature. Cryptoaescin contains C-
28-O-acetyl saponins, and β-aescin contains C-22-O-acetyl saponins, whereas
α-aescin is a mixture of crypto- and β-aescin (Dudek-Makuch and Studzińska-
Sroka 2015). Horse chestnut seed extract is widely used for the treatment of
peripheral vascular disorders, and in cosmetics (Ćalić-Dragosavac et al. 2011).
Saponin content in plants depends on many factors and varies not only
between plant species, but also between the parts of individual plants (Table
1). Cheok et al. (2014) reviewed saponin content in different plant parts
including seeds, roots, leaves, bulbs, fruits, stems, pericarps, tubers and
flowers. Phrompittayarat et al. (2011) analyzed and compared the total saponin
content in shoots, lower parts, roots, leaves, upper stems, and lower stems of
Brahmi plant collected in month 4 of the summer season, and found that the
percent content of saponin was highest in the leaves, followed by the shoots.
Szakiel et al. (2011) also reported that the amount and composition of
saponins depend on the light, temperature, humidity, local geoclimate and soil
fertility, as well as the season of the year and the cultivation techniques used.
For soil remediation, saponins should be used at a relatively high
concentration; therefore, nonfood saponin sources are more important than
edible sources. Saponin from Quillaia saponaria, Sapindus mukorossi, tea
saponin and aescin from Aesculus hippocastanum L. have predominantly been

tested for this purpose.
2. SURFACE ACTIVE PROPERTIES OF SAPONINS

Saponins, due to their amphiphilic nature, are surface active compounds
with detergent, wetting, emulsifying and foaming properties. Because of their
natural origin, saponins are termed plant-derived biosurfactants. Saponin
extracts can vary greatly in composition, mostly due to marked variability in
the raw material and differences in extraction protocols (Wojciechowski
2013). The purity of saponin extracts affects their properties, and is expressed
in terms of sapogenins content (Zhou et al. 2013). For soil remediation, the
most important properties of saponins are their ability to wet soil and to
generate stable foam, and also, their ability to decrease surface and interfacial
tension in aqueous solution, which is related to another important property,
their critical micelle concentration.
Table 1. Saponin content in different parts of selected plant species
Plant species Plant part Saponin content (% References

d.w.)
Allium nigrum L. roots 1.94 Mostafa et al. (2013)
bulbs 1.65
leaves 1.05
Quillaja saponaria branches (wood) 2.3-6.7 Copaja et al. (2003)
branches (bark) 6.6-13.3
trunk (bark) 6.5-15.8
trunk (hardwood) 2.7-9.3
twigs 5.8-14.4
Maesa lanceolata leaves (wildtype) 1.2 Theunis et al. (2007)
leaves (greenhouse) 4.9
roots (greenhouse) 1.5
Yucca schidigera stalk, roots 10 Oleszek and Hamed
Sapindus mukurossi fruits, roots 20 (2010)
Sapindus mukurossi fruit pericarp 14.2 Yang et al. (2010)
Sapindus officinalis aerial parts 22.4 Budan et al. (2014)
roots 8.2
Agave durangensis foliar tissue 0.58 Gonzalez-Valdez et al.
(2013)
2.1. Foam Generation and Stability
The factors that affect the ability of saponins to produce stable foam are
important to take into account because surfactants that show good and stable
foaming have good detergency, which is one of the properties that indicates
their effectiveness in removing pollutants from soil (Urum and Pekdemir
2004). Saponins are able to produce foam because of a combination of water
soluble sugar chains and non-polar aglycones in their structure. Usually foam
formation increases with increased saponin concentration. Chen et al. (2010)
found that the height of foam increased when the concentration of saponins in
extracts of Camellia oleifera increased from 0 to 0.1%. However, the foam
height of these saponins was moderate (4.6 cm) compared to synthetic
surfactants, i.e., sodium lauryl sulfate (SLS) (18.8 cm) and Tween 80 (13.6
cm). Urum and Pekdemir (2004) found that foam height was 4 cm at a saponin
concentration of 0.004%, whereas it increased to 22 cm when saponin
concentration was increased to 0.5%.
Foam produced by mechanical agitation is typically an unsteady
thermodynamic system, and the rate at which the foam weakens is used to
describe its stability (Mousli and Tazerouti 2007). The choice of the plant
from which saponin is extracted has a substantial effect on the properties of
the saponin foam (Böttcher and Drusch 2016): foams made from Quillaja
saponaria Molina, Gypsophila, Camellia oleifera Abel, and Aesculus
hippocastanum were more stable and dense than those from Glycyrrhiza
glabra. The more stable saponin foams were at 85% of their initial height after
1h (Böttcher and Drusch 2016). Saponin foam from Glycyrrhiza glabra may
have lower stability than the others because it does not reduce interfacial
tension as effectively, it incorporates only a small amount of liquid inside the
foam, and this saponin has poor solubility because the molecule has only a
small hydrophilic part.
A number of other factors also affect the stability of foams. First, foam
stability can depend on the ionic type of the surfactant. Foams from anionic
surfactants are usually more stable than from nonionic surfactants like
saponins (Lake 1989). Second, the ability of a saponin to produce foam also
depends on the number of sugar chains it has: saponins with a single sugar
chain have the best foaming properties, whereas those with two or three sugar
chains have inferior foaming properties (Oleszek and Hamed 2010). Third, the
formation and stability of saponin foams can be affected by environmental
conditions like pH and salinity. Usually, saponin foams are more stable at
more acidic pH. In acidic conditions the majority of the carboxylic groups in a
saponin moiety are non-dissociated and electrostatic repulsion is substantially

reduced. Although the effect of salinity on foam stability is not fully
understood, solutions with higher ionic strengths can increase saponin-foam
stability like solutions with lower pH (Böttcher and Drusch 2016).
2.2. Wetting
The wetting ability of saponins can be an important factor that helps

determine their effectiveness in treating contaminated soil. Better wetting
ability reduces the time it takes for a surfactant to penetrate the soil and make
contact with pollutants in the soil. Wetting is a complex phenomenon that
depends mainly on surface tension, surfactant concentration and the nature of
the surface being wet (Chen et al. 2010); of these three, wetting effectiveness
correlates most strongly with the ability of the surfactant to reduce surface
tension. Because saponins effectively reduce surface tension, they have good
wetting ability.
2.3. Surface and Interfacial Tension
As the surface and interfacial tension is lowered, the capillary forces that
hold a pollutant in the soil are reduced, which favors removal of the pollutant.
The ability of a surfactant solution to lower interfacial tension is especially
important for removing oil from soil (Abdul et al. 1990). The surface and
interfacial tension of good biosurfactants are less than 30 mN/m and 1mN/m,
respectively (Mulligan and Gibbs 1993). Saponins quite effectively reduce
surface and interfacial tension (Table 2); depending on the origin of the
saponins (commercial or extracted from plant tissues) and the method used for
measuring surface tension, saponins can lower the surface tension of water
from 72 mN/m to as little as 32 mN/m, in the case of saponin from Sapindus
mukorossi. Tea saponin also has a similarly high effectiveness in reducing
surface tension (Xia et al. 2009). In comparison to other biosurfactants,
saponins are less effective at surface tension reduction than rhamnolipids, but
more effective than tannic acid.
2.4. Critical Micelle Concentration (CMC)
As surface active agents, saponin monomers can aggregate into micelles at

specific concentration termed as CMC. The differences in CMC values for
saponins obtained from various sources can be caused by the degree of
saponin purity and variation in the composition of saponin from different
plants (Böttcher and Drusch 2016). Saponins can show better properties than
synthetic surfactants, e.g., saponins from E. arvense, and B. perennis, which
have low minimum critical micelle concentration values of 0.033 g/L and
0.076 g/L, respectively (Tmáková et al. 2015). Compared to other
biosurfactants, saponins have higher CMCs (Table 2).
Table 2. Selected surface active properties of saponins

and other biosurfactants
Biosurfactant type Biosurfactant Measurement Surface Interfacial CMC

production method tension tension (mg/L)
(mN/m) (mN/m)
Saponin1 commercial Du Nouy ring 39 6.0 1000
Saponin (S. mukorossi)2 extracted Du Nouy ring 32 n.d. 120
Saponin3 commercial Wilhelmy plate 39.3 n.d. 930
Saponin (S. mukorossi)4 extracted Du Nouy ring 41 n.d. 1000
Saponin (Q. saponaria)5 commercial Wilhelmy plate 45 n.d. 100-
200
Saponin (C. oleifera)6 extracted Du Nouy ring 43.5 n.d. 1814
Tea saponin7 commercial n.d. 32.9 n.d. 5000
Aescin from seeds of commercial Wilhelmy plate 44 n.d. 882
A. hippocastanum L.8
Tannic acid1 commercial Du Nouy ring 50 4.5 80
Rhamnolipids1 commercial Du Nouy ring 28 4.5 200
1 Urum et al. 2003, 2 Zhou et al. 2013, 3 Gusiatin 2014, 4 Mukhopadhyay et al. 2013,
5 Chen et al. 2008, 6 Jian et al. 2011, 7 Xia et al. 2009, 8 Pekdemir et al. 1999, n.d
– no data.
The ability of saponin to form micelles is strongly affected by

temperature, salt concentration and pH. An increase in temperature and salt
concentration led to a decrease in the CMC of Sapindus saponin. The CMC
was found to increase with an increase in hardness of water and an increase in
pH (Balakrishnan et al. 2006). Ribeiro et al. (2013) investigated the properties
of saponins from Agave sisalana and Ziziphus joazeiro. The highest reduction
in CMC values from Ziziphus joazeiro saponins was observed at neutral pH,
near room temperature (25-30 °C) and at saline concentrations between 2 and
4%, whereas the greatest reduction of the CMC of Agave sisalana saponins
was at pH 3-4 near room temperature, or at pH 10-11, temperatures about 55-
60 °C and saline concentration of 2 to 6%. Regarding Quillaja saponins from
different commercial sources, Mitra and Dungan (1997) reported CMC values
between 0.51 to 0.77 g/L at 25 °C; this value decreased to 0.35 g/L at 34 °C.
Although saponins form aggregates at specific concentrations, these
aggregates can differ in size and shape, depending on saponin origin. It has
been estimated that micelles formed by saponins from Saponaria officinalis
and soya bean are smaller than micelles formed by saponins from Quillaia
saponaria. The former can consist of only two molecules, the latter of 50
molecules or more (Oakenfull 1986, Oleszek and Hamed 2010). In addition,
the micelles of saponins from these plants have various shapes, from elongated
and filamentous (for saponin from Quillaia saponaria) to spherical (for
saponin from soya bean). The different shapes of the micelles can be caused
by the different structures of the aglycones in saponins of different origins.
With saponins extracted from quillaja bark and used at a concentration above
CMC (12.3 g/L), Mitra and Dungan (2000) showed that the saponin
aggregates have a monodisperse size distribution with an average diameter of
10 nm, and the size of the saponin micelles does not change with increased
concentrations (up to 40 g/L).
3. SAPONINS IN SOIL REMEDIATION

3.1. Heavy Metals
Heavy metals pose a serious environmental problem due to their

occurrence, persistence, tendency to bioaccumulation and toxicity in soil
(Alloway and Ayers 1999). As, Pb, Hg, and Cd are ranked 1st, 2nd, 3rd, 7th,
respectively, out of 275 substances on the Priority List of Hazardous
Substances (ATSDR 2015). The most commonly identified metals in
contaminated soils are Pb, Cr, Cd, Zn and Ni (USEPA 2002). Excessive heavy
metal concentrations need to be removed from soil to reduce their negative
environmental impact (Babula et al. 2008).
Soil washing is highly effective for removal of metals from contaminated
soils and is frequently used in soil treatment because this method can rapidly
clean a contaminated site, reduces or eliminates long-term metal mobility, and
is cost-effective (GOC 2003). Selection of proper washing agents influences

the efficiency of metal removal. Dermont et al. (2008) classified washing
agents for removing metals from soils into five groups: (1) acids; (2) salts and
high-concentration chloride solutions; (3) chelating agents; (4) surfactants; and
(5) reducing or oxidizing (redox) agents. Chemically enhanced soil washing
can be attractive if the chemical reagents have some or all of the following
characteristics: they can be recycled or detoxified, or they are not hazardous.
However, not all synthetic washing agents are safe. Thus, there is increasing
interest in biosurfactants because they offer an environmentally-friendly
option. In comparison with synthetic surfactants, biosurfactants often have a
larger molecular structure and more ligand groups, which endows them with
extraordinary surface activity for removal of mixed pollutants, i.e., heavy
metals and hydrophobic organics (Sachdev and Cameotra 2013).
Most studies with saponins have been carried out in laboratory batch tests.
The efficiency of soil washing with saponins varies, depending on many
factors like soil type, the concentration and chemical form of the metal, and
operational conditions (saponin origin and its concentration, pH, extraction
time, soil to liquid ratio, number of washings, etc.). The literature contains
reports on the performance of saponins in metal removal from different soils,
sludges and fly ashes.
3.1.1. Saponins for Soil Treatment

Saponins have been tested for treatment of different soils contaminated
with metals (Cd, Cr, Cu, Mn, Ni, Pb, Zn) and metalloids (As).
Hong et al. (1998) used aescin, commercially produced from seeds of the
horse chestnut tree (Aesculus hippocastanum L.), to remediate loam soil
artificially contaminated with Cd and Pb. A 3.4% aescin solution was most
effective for removal of Cd at pH 7.8 (41%) and removal of Pb at pH 2.8
(25%). This indicates that the carboxylic and saccharide moieties of aescin
may have higher binding capacities and selectivity for Cd than Pb. Therefore,
aescin is advantageous in remediation of Cd-contaminated soils at neutral pH,
which can minimize the risk of soil damage.
Hong et al. (2002) found that commercial saponin was a feasible choice
for remediation of soil in Japan that had been contaminated by heavy metals
for more than two years. Three soils (clay loam, loam, sandy clay loam) were
subjected to batch washing. Soil treatment with saponin was considered to be
suitable at a concentration of 3%, and an extraction time of 6 h. Using saponin
at pH 3 enabled nearly complete metal removal from soil. However, to
minimize the level of saponin sorption, the soil washing should be performed
above pH 3. Under alkaline conditions, metal removal efficiency was low due
to competition of sodium and heavy metals for saponin, leading to Na-saponin
complexes. Most of the heavy metals that were removed came from the
exchangeable and carbonate fractions, and only some from the Fe–Mn oxide
and organic matter fractions.
Chen et al. (2008), used kaolin as a model soil component and
contaminated it with Cu (450 mg/kg) and Ni (140 mg/kg). They found that
high efficiency of metal removal (83-85%) can be achieved at a relatively low
saponin concentration (0.2%), a kaolin dosage of 20 g/L, a pH 5-8 and a single
washing at room temperature. In addition to saponin concentration and
solution pH, soil dosage is also an important factor affecting metal removal.
Too high a soil dosage can cause more flocculation and sedimentation during
soil washing. As a result, the surface diffusion of micellar saponin aggregates
through the interparticle pores in the soil is limited, which diminishes metal
removal. According to the authors, metal removal by saponin at a
concentration above the CMC proceeds in a three-step mechanism: (i)
dissociated saponin micelles are sorbed onto soil particles, (ii) these adsorbed
saponin molecules compete with presorbed heavy metals for binding sites on
soil, which breaks down metal-ion pairs due to a high gradient of tension and
saponin film pressure, and (iii) hemimicelles, in the form of metal-surfactant
complexes, desorb into solution and aggregate into micelles.
Gusiatin and Klimiuk (2012) found that the efficiency of commercial
saponin (Sigma-Aldrich) for soil remediation strongly depended on soil type.
After a single washing with 3% saponin at pH 3, the highest efficiency of
metal (Cu, Cu, Zn) removal was obtained in loamy sand (82-90%) and loam
(67-88%), whereas the lowest was in silty clay (39-62%). Using different soils,
the authors demonstrated the following advantages to soil washing with
saponin: (i) although multiple soil washings were needed, saponin effectively
removed these metals from soils with high content of silt and organic carbon,
(ii) metal mobility was considerably decreased, which eliminates the problem
of uncontrolled metal migration in the environment.
Apart from commercial saponins, soil washing was also performed with
saponins extracted from soapberry. Maity et al. (2013) proposed using such a
natural plant biosurfactant for remediation of soil in Taiwan that had been
contaminated with Ni, Cr and Mn for over 10 years. The efficiency of metal
removal was the highest at pH 5 and it increased as saponin concentration was
increased from 0.015 to 0.15 g/L. Although Ni and Cr concentration in soil
were not high (151 and 70 mg/kg, respectively), the efficiency of Ni removal
(99%) was higher than that of Cr removal (73%) at a saponin concentration of
0.15 g/L and pH 5. Removal of Mn, which was present at the highest
concentration in the soil (476 mg/kg), was the least efficient (25%). These
results indicate that, although the extraction time is relatively long (24 h),
saponin from soapberry is able to remove heavy metals better than commercial
saponin.
Some researchers have also tested saponins for removal of arsenic (As)
from contaminated soil. As is a ubiquitous metalloid element that has attracted
considerable attention in recent years because its toxicity is of concern to
humans. The speciation forms and negative charge of As that is present in soil
make it different from cationic metals. As is retained in the soil mostly by
hydrous oxides of Fe(III) and Al(III), thus it is interesting to know if saponins
are able to remove negatively charged As ions.
Mukhopadhyay et al. (2013) used saponin extracted with water from the
pericarp of soapnut fruit for removal of As(V) from lightly (22.6 mg/kg) and
heavily (52.5 mg/kg) contaminated soil. With both column washing
procedures (down-flow mode and top-flow mode), the cumulative efficiency
of As removal at a 1% saponin concentration was similar, but removal from
lightly contaminated soil (~45%) was more efficient than removal from
heavily contaminated soil (~35%). Analysis of the FT-IR data suggested that
the saponin did not interact chemically with As, indicating that it was possible
to reuse the biosurfactant. Thus soapnut saponin can be considered for use as a
soil washing agent for removing As from soils, including those with a high Fe
content.
Gusiatin (2014) tested commercial saponin for removal of As from soils
from a former mining and smelting area in Zloty Stok, lower Silesia (Poland).
As content varied from 2041 to 4294 mg/kg. The acidic nature of the saponin
favored decomposition of Fe oxides and As release. After triple washing with
3% saponin at pH 3.44, the content of As in soils decreased by 52-63%.
Saponin reduced As toxicity in the soils by efficiently removing As(V) and
As(III). However, efficient As removal needs a relatively long extraction time
(24 h) (Gusiatin 2015). Nevertheless, saponin can be attractive candidate for
As removal from soil at a field-scale.
3.1.2. Saponin for Sludges and Fly Ashes Treatment

With the rapid development of industry, large quantities of different waste
are produced every year. Because these waste usually contain different
pollutants, especially heavy metals, they should be pretreated before their
disposal or further management. The literature indicates that there have been
attempts to use saponins for treatment of contaminated sludges or fly ashes.
Gao et al. (2012) used commercial saponin at concentrations 1-50 g/L and
pH 2.5-6.5 to remove metals from artificially contaminated sludge collected
from an industrial water treatment plant. Although the best results at batch
conditions were obtained at saponin concentration of 50 g/L, the authors
postulated that using a high saponin concentration (50 g/L) could produce
colloidal precipitation due to adsorption of saponin onto sludge. In addition, at
saponin pH higher than its pKa (4.6), the removal efficiency of metals was
low, whereas at pH lower than the pKa, it abruptly increased. The efficiency of
Ni and Cr removal with saponin at a pH lower than 3 was reduced due to
saponin sorption onto sludge. In column experiment, saponin at concentration
of 30 g/L and pH 3 facilitated mobilization of metals selectively and the
leaching behavior depended on the characteristics of metals. After 12 washing
volumes (total volume of saponin solution was 74 ml), the removal efficiency
of metals (originally present at total concentrations of 1210 mg/kg (Pb), 716
mg/kg (Ni) and 984 mg/kg (Cr)) decreased in this order: 73.2%, 64.2% and
56.1%, respectively.
In contrast, Kiliç et al. (2011) used commercial saponin from Quillaja
bark in remediation of tannery sludge highly contaminated with Cr (8041
mg/kg). The effects of various factors like time, concentration of saponin, pH,
and temperature on Cr removal were studied. The maximum Cr removal
(24.2%) was obtained by performing double washing process for 6h with 5%
saponin at pH 2 and at 33 °C. A relatively low removal efficiency resulted
from the fact that tannery sludge acted like aged contaminated soil and Cr was
tightly bound to colloidal particles and organic matter in sludge, which
affected its mobility.
An extraction process with saponins for removing heavy metals from
municipal solid waste (MSW) incinerator fly ashes was evaluated by Hong et
al. (2000). The fly ashes varied significantly in chemical composition and
concentration of Cu, Pb, Zn and Cr. Three types of saponins were tested at
concentration of 3.75% and pH range 4-9: saponin M (mixture of quillaja bark
and saponin grass), saponin Q (from quillaja bark) and saponin T (from tea
seed). The efficiency of metal extraction depended on metal and saponin type.
The best results were obtained at pH 4-5. Saponin T extracted 100% of Pb,
which was initially present in the fly ash at a concentration of 6900 mg/kg. All
of these saponins extracted 20-45% of Cr and 50-60% of Cu from the fly
ashes.
3.2. Heavy Metals and Organic Pollutants
The contamination of soils with mixed pollutants (heavy metals and

hydrophobic organic compounds) is a widespread environmental problem. For
example, according to reports of the US Environmental Protection Agency
(USEPA), more than 67% of contaminated sites are found to contain heavy
metals and organic pollutants simultaneously (USEPA 2004). An important
source of pollutants in soil is hazardous waste. The USEPA National Priority
List indicates that 40% of the hazardous waste sites are co-contaminated with
organic and inorganic pollutants (USGAO 2010). The co-occurrence of
polycyclic aromatic hydrocarbons (PAHs) and heavy metals exacerbates their
toxicity compared to individual pollutants (Shen et al. 2006; Soltani et al.
2015). Thus, much attention has recently been paid to the simultaneous
removal of heavy metals and organic pollutants from soils, although co-
occurrence of these pollutants can complicate remediation process.
In soil washing technology, three approaches have been tested for
simultaneous removal of mixed pollutants: single surfactant washing,
sequential washing with different agents and a combination of surfactants and
chelating agents (Cao et al. 2013). The literature indicated that saponin could
effectively remove mixed pollutants from soil.
Song et al. (2008) found that commercial saponin had potential for
simultaneous removal of phenathrene and Cd from soil. At concentration of
0.375%, saponin removed 87.7% Cd and 76.2% phenanthrene. Saponin is able
to simultaneously remove different pollutants without competition for binding
sites because the metals bind at different locations on the saponin molecules
compared with organic pollutants. In Song et al. (2008), phenanthrene was
partitioned into saponin micelles, while Cd was complexed with the external
carboxyl groups of saponin micelles.
Wang et al. (2015) investigated the effect of different washing agents (tea
saponin, rhamnolipid, alkyl polyglucosides and sodium dodecyl benzene
sulfonate) on solubilization and distribution of pyrene and Cd in co-
contaminated soil. Tea saponin at concentration of 40 mg/L was more
effective in solubilzation of these pollutants than other washing agents. With
adding of tea saponin, changes in pollutant distribution in soil were observed.
Cd was transformed from Fe-Mn oxides and carbonate fraction into
exchangeable fraction, whereas pyrene was transformed from associated
fraction to bioaccessible fraction. These investigations revealed the presence
of Cd in co-contaminated soil promoted solubilization of pyrene.
Some investigators showed a positive effect of using saponin solution

mixed with other additives to enhance removal of combined pollutants from
soil. Ye et al. (2015) demonstrated that two successive washings with a
mixture of 5 ml/L peanut oil and 0.5% tea saponin were effective to remove
organic pollutants and heavy metals from e-waste contaminated soil. Tea
saponin promoted desorption of organic and inorganic pollutants from soil into
the aqueous solution, which next moved from aqueous phase into the more
hydrophobic peanut oil, which served as a sink for solubilized pollutants. The
mixture of tea saponin and peanut oil removed 94.6% PBDEs, 97.1% PCBs,
95.1% PAHs, 83.5% Pb and 87.1% Ni.
Similarly, Cao et al. (2013) investigated simultaneous removal of Pb, Cu
and PCB from soil with a mixture of saponin and chelant S,S-
ethylenediaminedisuccinic acid (EDDS). Combination of saponin and EDDS
had a synergy effect on pollutant removal and would be a promising
alternative for remediation of co-contaminated soil. With adding a mixture of
10 mM EDDS and 0.3% saponin, the efficiency of Pb, Cu and PCB removal
from soil was 99.8%, 85.7% and 45.7%.
4. SAPONIN RECOVERY
In order to improve the cost-effectiveness of soil remediation, the recovery
and reuse of surfactants after soil washing is necessary. For example, the price
of commercially available saponin from quillaja bark with sapogenins content
≥10% is 174 euro per 100 g, while with sapogenins content of 20-30% it is
nearly four times higher (Sigma-Aldrich). After washing of soils or sediments
contaminated with heavy metals, the leachate contains mainly these pollutants.
Thus, saponin recovery is based on metal removal from the solution. For that
purpose physico-chemical methods can be used, including precipitation and
adsorption. However, precipitation is more commonly used than adsorption.
Gao et al. (2012) used precipitation to remove heavy metals from saponin
solution after washing of industrial sludges. Metal recovery was pH-dependent
and the optimal pH was 10.9, at which metal removal efficiency was 89.7%
for Pb, 91.1% for Ni and 99.1% for Cr. At pH above 11.5, hydroxide
compound of Pb and Cr started to redissolve due to amphoteric nature of these
metals.
In Gao et al. (2013) it was reported that saponin recovery becomes lower
with increased the recycling times. The recovery efficiency of saponin after
the first sludge washing was 87-94%, whereas it decreased to 62-75% after the
second sludge washing. The loss of saponin in subsequent washings and

recoveries may be attributed to saponin adsorption on sludge surface and/or
hydroxide compounds surface. However, the recovered saponin has shown
satisfactory ability to remove heave metals from sludge.
Precipitation method was considered by Hong et al. (2002) to be useful in
recovery of saponin after soil washing. At pH 10.7, the precipitation of heavy
metals was 86% of Cd, 80% of Cu, 90% of Pb and 91% of Zn. The recovered
saponin was able to remove heavy metals from soil with slightly lower
efficiency than the original saponin. This indicates the possibility of
subsequent utilization of recovered saponin.
Similarly, Mukhopadhyay et al. (2015) used ferric chloride to precipitate
As from soapnut solution after soil washing. Soapnut solution can be
recovered from the wash effluent with low dosage of ferric chloride (8-10
mg/L) and at pH 8. Under these conditions up to 87% of As was removed
through coagulation, flocculation and precipitation. Ferric chloride is found to
be effective precipitating agent for As. Thus, Gusiatin (2015) used FeCl3 to
modify properties of mineral sorbent in form of clinoptilolite to recover
saponin after washing of soils contaminated by arsenic ore processing.
Depending on the As content in soil, concentration of As removed with
saponin varied between 9.5 an 54.6 mg/L. Modified clinoptilolite at a dosage
of 300 g/L removed ≥80% of As from saponin effluents. Although the reused
biosurfactant did not extract As as effectively as the original solution, pH
adjustment improved its effectiveness. In the case of As removal, the pH of
recovered and reused washing agent can be more important than its
concentration (Kim and Kim 2011).
5. NEW TRENDS OF USING SAPONIN

IN SOIL REMEDIATION
Researches on saponin application in soil remediation are still being

developed. The new trends in soil remediation with saponins include: (i)
saponin foam technology, (ii), saponin-DES washing and (iii) saponin
enhanced phytoremediation.
5.1. Saponin Foam Technology
Surfactant foam (SF) technology has been suggested to achieve better

mobility control in porous media and to improve removal efficiencies of
pollutants (Wang and Mulligan 2004). Foam is a two-phase system where the
mass of gas or air cells is dispersed in a liquid and separated by thin liquid
films called lamellae. Sometimes gas injection directly into the porous
medium is also possible (Atteia et al. 2013). Surfactant foams to be used in
soil remediation should have proper quality (as foam volume) and stability
(ability of the foam to resist bubble collapse) (Wang and Mulligan 2004).
Different surfactants display different foamabilities. However, saponins
display sufficient foaming properties to be used in SF technology.
The positive effects of SF technology on metal removal from soil have
been shown with rhamolipids (microbial biosurfactants) (Mulligan 2005).
Although studies on saponin foam technology are not numerous, the
preliminary results are very promising. The feasibility of using saponin from
soapberry for removal of heavy metals from contaminated industrial soil (6511
mg Cu/kg, 4955 mg Pb/kg, and 15090 mg Zn/kg) was investigated by Maity et
al. (2013). For soil washing, foam fractionation and soil flushing were used.
According to the authors, a suitable foam stability for soil treatment needs
higher flow rate and higher saponin concentrations. At higher temperatures
(>40 °C), the foam is unstable and easily collapses, resulting in a lower
removal rate. The research indicated that foam technology was more efficient
for the removal of heavy metal than soil washing with aqueous saponin
solution. The efficiency removal of Cu, Pb and Zn by foam was 95%, 98% and
56%, respectively, at the following operational conditions: 0.015% saponin,
pH 4.0, 1.0 ml/min. flow rate and 30 °C. Foams have higher viscosity, and
creating a sharper injection front increase efficiency of pollutant removal
compared to aqueous solution (Atteia et al. 2013).
Apart from solutions and foams, surfactants can be used for soil washing
in a form of colloidal gas aphrons (CGAs). CGAs is a system of spherical or
spheroidal microbubbles with diameters above 25 μm suspended
homogenously in a liquid matrix. Thus, CGAs and regular foams differ in
bubble morphology and are more stable than regular foams. Mukhopadhyay et
al. (2015) evaluated performance of the CGAs in comparison to solution to
remove As from soil in a column reactor using saponin from Sapindus
mukorossi and sodium dodecyl sulfate (SDS). The efficiency of As removal
from the soil columns using soapnut-CGAs was 70% and it was higher by
15% than SDS-CGAs. Although the efficiency of CGAs and saponin solution
was comparable, the advantage of CGAs is economical, as this system

contains 35% of air by volume, thereby requires less biosurfactant.
5.2. Saponin-DES Washing
There are new possibilities of application of natural compounds based on

deep eutectic solvents (DESs) and mixing them with surfactants to enhance
soil washing efficiency. A DES is a mixture of two or more compounds that
has a melting point lower than that of either of its components (Hayyan et al.
2010). In practice, the DESs are obtained by mixing hydrogen bond donor
with a salt and water.
Mukhopadhyay et al. (2016) demonstrated a synergistic effect of using the
DESs and saponin extracted from soapnut fruit pericarp by water to remove Pb
from spiked soil. To prepare the DESs, quaternary ammonium salt in form of
choline chloride was used, and four types of hydrogen bond donors (HBD),
i.e., fructose, sucrose, glycerol and ethylene glycol. Using 0.5% saponin, Pb
removal efficiency was about 30%. Fructose and sucrose used at concentration
of 10% were more effective HBD than glycerol and ethylene glycol removing
Pb up to 31% and 25%, respectively. Using DES-0.5% saponin mixtures with
fructose and sucrose donors, Pb removal efficiency increased up to 49-55%.
The efficiency increased up to 70% after increasing saponin concentration to
2% in the mixture with DES. These findings indicate that soil washing by
DESs and saponin solutions involves a Lewis acid–base interaction. Whereas
the soil has a deficiency of electrons, the organic carbon produces electron
donating points, thereby acting as an Lewis base, which donates an electron
pair, attracting and binding Pb, which acts as Lewis acid. Saponin, DES-
fructose and DEC-sucrose are source of H+ due to their acidic character. In a
DES-saponin mixture more H+ is introduced to soil. These H+ ions compete
with Pb for electrons, especially on the electron rich sites on organic carbon in
the soil. As a result, Pb ions can be attached with DES anions and dissociated
groups of saponin and removed from soil. The biological origin of natural
deep eutectic solvents and natural saponin make the process of soil washing
promising and sustainable.
5.3. Saponin Enhanced Phytoremediation
The success of phytoremediation depends mainly on the pollutant

availability to plant roots. Because some pollutants are quite tightly bond to
soil constituents, their accumulation by plants roots can be limited. In order to
increase phytoremediation efficiency, the solubility of pollutants can be
promoted by a number of chemical approaches, including the application of
chelators and surfactants. Phytoremediation combined with adding
biodegradable biosurfactants like saponins would be an effective in situ
method for removal of organic pollutants and heavy metals.
Tea saponin has potential to promote the uptake of different pollutants by
plants from soil. Xia et al. (2009) tested tea saponin to enhance uptake of PCB
and Cd from soil by Saccharum officinarium L. Application of 0.5% saponin
significantly increased PCB accumulation in roots and shoots, whereas
addition of 0.3% tea saponin increased Cd concentration by 96.9% in roots,
156.8% in stems and 30.1% in leaves. Cay (2016) used tea saponin to enhance
uptake of Cd by Amaranthus caudatus. With the increase of saponin dosage
from 1 to 3 g/kg, Cd accumulation increased in roots, stems and leaves.
According to the authors, phytoremediation enhanced with saponin could be
applied in catchment areas of urban cities. Wang et al. (2016) found that tea
saponin promoted Cd accumulation by Lolium multiflorum and stimulated
microorganism activity responsible for degradation of pyrene in soil.
Plant-derived saponins are important agents in soil remediation due to
their biodegradability, low toxicity and high selectivity for metal removal
compared to other washing agents. Use of plant biomass as source of saponins
can be a new manner of biomass management that should be extended in the
future. Application of plant extracts containing saponins can significantly
lower costs of soil remediation compared to commercial saponins. The
possibility of using saponins as liquids or foams increases the potential of their
application. After highlighting the usability of saponins for pollutants removal
from soil at laboratory scale, their application at field scale is necessary, not
only in soil washing, but also in other remediation technologies.
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Chapter 4
APPLICATION OF SAPONIN FROM SOAPNUTS

FOR REMEDIATION OF OIL
CONTAMINATED SAND
Meshari S. Almutairi, PhD

School of Civil Engineering and Surveying, University of Portsmouth,
Portsmouth, UK
ABSTRACT
The aim of this research was to evaluate the feasibility of using
saponin from soapnuts (Sapindus mukorrossi) for the removal of oil from
a large area of oil contaminated soil in Kuwait which is an aftermath of
the destruction of oil wells during the Iraqi invasion of Kuwait.
Experiments were carried out to assess the efficiency of soil washing
using a non ionic biosurfactant saponin under different conditions like
concentration of saponin, washing time, stirring speed, operating
temperature and mass/volume ratio. The optimum concentration of
saponin was found for the most effective removal of oil from the
contaminated sand by using artificial sea water keeping parameters such
as a temperature, soil/solution ratio, stirrer speed and washing time
constant. The results from this study indicates that saponin can be
appropriately used for its application as a biosurfactant for washing oil
contaminated sand.

Corresponding Author: Email: meshari.almutairi@ myport.ac.uk.
92 Meshari S. Almutairi
Keywords: soil remediation, soil washing, biosurfactant, saponin, soapnuts
1. INTRODUCTION
The term ‘Saponin’ is originated from the word ‘sapo’ in Latin language
which means soap, owing to the foam generating characteristic in aqueous
solution by stirring action. Saponins are glucosides with non-ionic
biosurfactant properties and foaming characteristics. They are extensively
distributed in the territory of plants (Roy et al., 1997; Tewari et al., 2011), like
vegetables, fruit pericarp, soybeans, peas, beans and herbs, whereas saponin
which are commercially produced are obtained largely from Sapindus
mukorrossi (soapnuts), Yucca schidigera and Quillaja saponaria. Saponin
solutions are amphiphilic glycosides in which the polar components include
sugars (pentoses, hexoses or uronic acids) which are connected covalently to a
non-polar group. Saponin’s structure (Figure 1) is in the form of lipid soluble
polycyclic aglycones connected to a single or multiple sugar chains and water-
soluble sugar residues (Hong et al., 2002). The aglycone element also known
as sapogenin can be either steroid (C27) or triterpene (C30). According to
Shiau et al. (2009) and Sharma et al. (2013) saponins consist of several
functional groups such as, OH, COOH, carboxylate, acetate group and esteric
band.
Source: Fisher Scientific, UK.
Figure 1. General chemical structure of saponin.

Application of Saponin from Soapnuts … 93
1.1. Classification
The classification of surfactants are made according to the type of head

group, i.e., cationic, anionic or non-ionic. Non-ionic surfactants are neutral
which result in lower affinity to electrolytes in the solution. With regard to soil
washing, if there are metal ions, salt, etc. present in the solution, an electrolyte
is introduced to the aqueous surfactant solutions for improving the efficiency
of oil removal from soil (Zhong et al., 2003). As such, the non-ionic
surfactants are able to prolong the contact time in the water bath. According to
Mulligan et al. (2001), anionic and non-ionic surfactants are more frequently
used in soil remediation as they are less expected to sorb onto surfaces of the
soil. Some of the benefits of non-ionic surfactants are that they have a high
solubility in cold water, low microbial toxicity and better wetting action (Kim
and Wang, 2003; Zhao et al., 2005). Cuypers et al. (2002) and Conte et al.
(2005) revealed that non-ionic surfactants are able to desorb hydrocarbon
compounds from soil.
1.2. Ecofriendliness and Applications of Saponins from Soapnuts
Saponin from soapnuts (Figure 2) are well known for its effectiveness as a
natural multipurpose detergent for cleaning laundry, dishes, floor and pets. It is
completely biodegradable hypoallergenic and be used for cleansing sensitive
skin and finds application in cosmetics. Now the demand for saponin is
booming since the awareness for green technology is escalating worldwide.
This research primarily focuses an alternate application of saponins in a green
technology for remediation of oil contaminated sand particularly in the case of
remediation of oil lakes in Kuwait.
Figure 2. Sapindus mukorossi (soapnuts).

1.3. Case Study of Oil Lakes in Kuwait
During the occupation of Kuwait by Iraq in 1990, Iraq’s armed forces

destroyed more than 700 oil wells. The resulting oil flowed out and formed a
large number of oil lakes. This also led to the contamination of soil, which has
been left untreated in the deserts of Kuwait for more than two decades now.
The untreated contaminated soil if left as such may lead to pollution of
underground watercourses and would be a threat to the environment and
neighboring population. So the necessity of developing a green technology for
remediation of soil was contemplated by the author. Oil contaminants being
hydrophobic in nature, the main purpose of using surfactants is to minimize
the hydrophobicity of the oil phase to such a level that it is wetted by the water
phase and separate itself from the surfaces of the soil. As such, surfactants are
employed in order to improve the surface actions of the surfactant/oil/soil
systems. Also, chemical surfactants are effective in reducing both the viscosity
of oil and the interfacial tensions of water and oil (Al-Sabagh, 2000; Liu et al.,
2004). However, there may not be true solubility of petroleum compounds in
water and it has been observed that surfactants can also increase pseudo
solubility of hydrophobic contaminants (Chu, 2003; Pekdemir et al., 2005).
For example, in the presence of saponins, PAH solubility increases thus
making them more available to degrading bacteria (Soeder et al., 1996). The
removal of organic contamination from solid matrices by treatment with
surfactant is considered as an affordable and comparatively rapid procedure
and can treat contaminants in large scale. The environmental impact of
surfactant has increased the awareness of using biosurfactant as
environmentally friendly cleaning product. Nevertheless, traditional synthetic
surfactants were used by Hirata et al. (2009), which has caused difficulties in
downstream processes due to their toxic characteristics and non-
biodegradability even after 8 days. Consequently, owing to lesser toxicity,
better biodegradability and stability under the environmental conditions (e.g.,
temperature and pH) and generally less cost, biosurfactants are more suitable
for environmental applications than their synthetic counterparts (Muthusamy
et al., 2008).
1.4. Soil Washing as a Remediation Strategy
Soil washing has been extensively researched for the remediation of soils
and sands contaminated with crude oil constituents (Deshpande et al., 1999;
Vreysen and Maes, 2005; Santa Anna et al., 2007). Soil washing technology
uses water, occasionally mixed with solvents or surfactants in specifically
designed mechanical processes to clean soils. Stirring has been proved to
enhance the efficiency of traditional methods using hot water for elution (Feng
and Aldrich, 2000). It has been proven in the literature that biosurfactants play
a significant role in enhancing the soil washing process. This hypothesis led to
the selection of saponin as the biosurfactant, using a decision-making table.
1.5. Method Development for Extraction of Saponin
Any process design demands to select the most cost-effective raw

materials for its unit operations during the pilot and commercial scale.
Biosurfactant being one of the crucial raw material used in this research, the
selection, availability and extraction from natural source was a challenging
task. Purchasing the reagent grade saponin in its purest form is very expensive
which is not economically feasible for a commercial scale soil washing.
Therefore, this research targeted to reduce the surfactant cost and ultimately
the total cost of soil washing by developing a method for extraction of saponin
(biosurfactant) from the soapnuts. Reagent grade saponin obtained from
(Fisher Scientific, UK) costs around £113 per 100 g. While, extracted saponin
(less purity) cost as low as £1 per kg. Hence extracted crude saponin powder
was characterized by a single step of analysis by fourier transform infra-red
spectroscopy (FTIR) thereby eliminating the need of further expensive and
exhaustive purification steps. The outcome of this research is expected to
provide a better understanding of the removal of the oil residue under specific
conditions as well as to provide useful information about the application of
saponins in soil washing techniques in remediating hydrocarbon contaminated
soils.
2. MATERIALS
2.1. Saponin
Dry fruits of Sapindus mukorossi (Lot# HAUS57/30185-1) commonly

known as soapnuts were purchased from ISKA GmbH, Germany. Laboratory
reagent grade standard saponin (from Quillaja saponaria) was purchased from
Fisher Scientific, UK. Absolute ethanol was purchased from Aldrich, Tryptone
soya agar (CM0131 from OXOID, UK). Defibrinated blood from sheep was
used for the preparation of blood agar plate obtained from Saudi prepared
media laboratory (LTD). Distilled water was used for preparation of artificial
seawater for soil washing.
2.2. Contaminated Soil Sampling
The greater Al-Burgan oil field, in the desert southeast of Kuwait was
selected for sample collection. Extreme care was taken in selecting appropriate
section of the site as areas in proximity to the oil fields have been reported to
contain unexploded ordnance. The soil samples were collected from Burgan
oil field using a hand shovel to gather about 3 cm of oily sludge, and oil
contaminated soil was collected upto a depth of approximately 30 cm, below
the surface level of oily sludge. Subsequently, the samples were placed into
plastic containers and transported to the laboratory for further analyses and
soil washing experiments.
3. METHODS
3.1. Preparation of Oil Contaminated Sand
The sample of oil contaminated sand was obtained from Kuwaiti oil lake.
One kg of soil sample was screened manually through 13.5 mm sieve for 2
min, to remove twigs, stones or any other oversized material.
3.2. Characterization of Oil Contaminated Sand
3.2.1. Total Petroleum Hydrocarbons (TPH)

The total petroleum hydrocarbons of the petroleum contaminated sand
before and after each soil washing experiment was determined by the standard
EPA method 3546.
3.2.2. Electrical Conductivity

The oil-contaminated soil was mixed with distilled water to measure the
electric conductivity (in units of μS/cm) with a ratio of 1:5 weight to volume.
The conductivity of the oilcontaminated soil was measured by using a Jenway

4010 conductivity (BS EN 13038: 2011).
3.2.3. pH
The pH of oil contaminated sand was measured based on BS ISO (10390:
2005). 30 g of oil contaminated sand sample was dried for 24 h in a drying
cabinet at 40°C (Lec, UK), reweighed and placed into beaker containing 150
ml of distilled water and mixed for 10 min by mechanical stirrer (Cole-
parmer).
3.3. Preparation of Soapnuts and Extraction of Saponin
The preparation and extraction of crude saponin was carried out as

described by Almutairi and Ali (2015). The dark brown coloured soapnuts
were sorted and outer pericarps were separated from the seeds manually. Dry
soapnuts (Sapindus mukorossi) were ground to powder and was extracted with
95% ethanol at a ratio of 1:10 (w/v). The suspension was agitated on a hotplate
with magnetic stirrer 3 hours at 45°C and centrifuged at 12,000 rpm for 30 min
at 25°C to obtain the supernatant which was then freeze dried to get fine
powder. The phytochemical tests on the extracts were carried out according to
standard methods as described by Tewari et al. (2011) and Sotheeswaran
(1988). The extracts of soapnuts were screened for saponins along with
controls by phytochemical tests like foam test and hemolysis test.
3.4. Characterisation of Saponin Extract by Fourier Transform

Infra-Red Spectroscopy (FTIR)
Dry powder of the extract obtained was mixed with KBr salt, using a
mortar and pestle, and compressed into a thin pellet. Infrared spectra were
recorded as KBr pellets on a Jasco Vacuum FT/IR 6300 between 500-4000
cm-1. Infrared absorptions were recorded for the direct detection of
triterpenoid saponins as described in Kareru et al. (2008).
Source: Almutairi (2016).
Figure 3. Experimental set up for soil washing.
3.5. Saponin Solution
Saponin solutions were prepared by weighing 0.01, 1, 5, 10 and 20 g of

saponin powder extracted from Sapindus mukorossi and adding it carefully to
1litre each of artificial seawater in separate glass beakers and stirred for 10
min at 200rpm to ensure complete dissolution.
3.6. Optimization of Saponin Concentration for Soil Washing
Several experiments were conducted for the optimization of conditions

like type of water used, temperature, time for washing, stirrer speed and mass:
volume ratio. The optimum parameters chosen for soil washing were washing
temperature of 50°C for 20 min at a stirring speed of 1000 rpm at a soil to
solution ratio of 1:3 (w/v) in artificial seawater. Keeping these parameters
constant, soil washing experiments were conducted at different concentrations
of saponin ranging from 0.001 to 2 wt%. All experiments were repeated 5
times unless mentioned otherwise. The soil washing experiment was carried
out mixing 50 g of the oil contaminated soil and 150 ml of saponin at each
predefined concentration in a 250 ml glass beaker. The beaker was placed in a
water bath maintained at 50°C and mixed with a mechanical stirrer (CP Cole-
Parmer) under the above mentioned conditions as shown in Figure 3.
3.7. Surface Tension
Surface tension of the oily wastewater at various concentration of saponin

were determined using a Du Nouy tensiometer (Type: K20. KRÜSS, Korea)
using platinum ring method. Each experiment was conducted by using 50 ml
of the sample in a 150 ml beaker. During the process, the distance between the
liquid surface and immersed ring was kept approximately 5 mm to insure the
position of the ring just below the liquid surface. The preprogrammed software
in the tensiometer measured the surface tension of the surfactant solutions.
4. RESULTS AND DISCUSSION

4.1. Soil Characteristics
Chemical properties were investigated by determining TPH, EC, pH and

metal contents of Kuwaiti oil contaminated sand. The outcome demonstrated
that the average value of TPH was 367,000 mg/kg (SD = 4,700, n = 3). The
present preliminary study shows a high average value of electrical
conductivity (EC) (2,455 μS/cm) (SD = 440, n = 3), the existence of ions
could be caused by using seawater to extinguish fires from oil well. Moreover,
the average of pH (7.9) for the contaminated sample was found slight higher
than the permissible level of pH 7.5 (KEPA, 2014). As illustrated in the Table
1, the results indicate the presence of some elements such as Ba, Cr, Fe, Ni,
Pb, Cd and Ag.
Soil washing was performed to determine the optimum concentration of
surfactant for maximum washing efficiency. This range of saponin
concentration has been proved to effectively remove oil contamination from
practical point of view. The result proves that the percentage of oil removal
from Kuwait oil residue specimens improved steadily by increasing the
concentration from 0.001, 0.1, 0.5, 1 and 2 wt% were about 35, 44, 64, 67 and
69%, respectively as shown in Table 2.
Table 1. The concentration of contamination in Kuwait oil contaminated

sand (n = 3)
Metals Average concentration of metals KEPA limit

(mg/kg) (mg/kg)
Barium (Ba) 0.78 10
Chromium (Cr) 0.52 5
Iron (Fe) 8.10 5
Nickel (Ni) 0.43 10
Lead (Pb) 0.70 5
Cadmium (Cd) 1.10 1
Table 2. Removal of oil contamination on increasing concentration

of saponin
Saponin concentration (wt %) 0.001 0.01 0.5 1 2

Removal of TPH% 35 44 64 67 69
However the 0.5% is found to be most economically feasible without

much compromise in washing efficiency and hence can be chosen for small
scale as well as large scale soil washing purposes. These experiments provide
a clear notion that there is no significant difference between removal of oil
from the sand where 0.5% and 1% saponin has been used. Moreover, it can be
concluded that concentration of saponin at 0.5 wt% will make a significant
difference in terms of cost of saponin and therefore it is not reasonable to use
more than 0.5% saponin.
It can be seen from Figure 4 that the decaying curves of TPH
concentration with various concentration of saponin showed logarithmic curve
trend.
4.2. Surface Tension
The surface tension was measured during this study to select and evaluate
the optimum concentration of saponin required to achieve high efficiency
removal of the oil residue. The surface tension of the oily wastewater
supernatants was measured at 20°C by a Du Nouy tensiometer. The CMC
value was determined by plotting surfactant tension versus log of surfactant
concentration. CMC was found at the intersection point on the slope where
surface tension shows linear decline meets the baseline of minimal surface
tension. Based on the literature, the most effective removal of crude oil was
registered at concentration below their CMC values, and does not exhibit any
improvement in removal of the oil residue at higher concentration. In order to
measure the surface tension, washing process was conducted using artificial
sea water at 50°C for 20min at a stirring speed of 1000 rpm at a soil to solution
ratio of 1:3 (m/v) at saponin concentrations like 0.001, 0.1, 0.5, 1, 2 and 3
wt%, as shown in Figure 5. This experiment was repeated three times for each
concentration to ensure reproducibility.

Note: The data are expressed as mean ± SD (R2 = 0.90; n = 5).
Figure 4. Mean of oil residue with different concentration of saponin.
The outcome of Figure 5 illustrates that the surface tension test of 0.001,
0.1, 0.5, 1, 2 and 3 wt% for various concentrations of saponin solution were
53.2, 41.1, 37.2, 36.0, 35.0 and 35.5 N/m, respectively. Furthermore, surface
tension between air and oily wastewater was measured as 77.2 N/m. Based
upon findings, as the surface tension between oil residue and saponin solution
is reduced, the attraction force holding the oil residue and sand is reduced as
well. It seems that the reduction of surface tension demonstrates the ability of
saponin to remove oil residue from sand. The proportion of micelles present at
the surface of a liquid depends on their concentration. As such, surfactants
occupy the surface of the liquid at low concentrations, this study found that
only slight change was detected in surface tension at low concentrations of
saponin, however Figure 5 shows that additional saponin decreases surface
tension. This suggests that the removal of crude oil was caused by
mobilization that took place due to the drop in the surface tension. As such,
when a solution of saponin is introduced, the surface tension decreases until
the CMC value is achieved at concentration of 0.5 wt% and remains constant
with little change beyond this point.
Figure 5. A conventional plot of the surface tension versus the logarithm value of the
surfactant concentration at ratio 1:3 (m/v).
CONCLUSION
The use of saponin in soil washing technology for remediation of
contaminated sites is presented as an attractive option because of its
biodegradability, ecological safety and environmental acceptance. Naturally
occurring saponin derived from soapnuts finds its application for remediation
of soil contaminated with hydrocarbons. The optimum concentration of
saponin for the most effective removal of oil from the contaminated sand is
found to be 0.5% (w/v) on using artificial sea water at a temperature of 50°C,
soil/solution ratio of 1:3 (m/v), a stirrer speed of 1000 rpm and washing time
of 20 min. The oil contamination removal efficiency increases with increase in
saponin concentration. It can also be concluded that saponin solution may have
no impact on the oil removal properties at concentration higher than their
critical micelle concentration level. Therefore it is indicated that saponin from
soapnuts can be used for its application as a biosurfactant for cases such as of
oil lakes in Kuwait.
ACKNOWLEDGMENTS
The author extends his gratitude to Ministry of Higher Education, Kuwait,
for funding this research. Thanks are also due to KOC and KNPC for their
support in soil sampling.
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75.
BIOGRAPHICAL SKETCH
Dr. Meshari Almutairi
If life may be compared with a race without a definite destination, every

goal one has attained will only begin the pursuit of another. By doing so, we
enrich our own lives, and those of others.
However, as an engineer at thirty years of age, I need to do a little
persuasion to myself to start an academic life again. It might be a process full
of difficulty and challenges. But my character and my past experience have
convinced me that my life would be better with these challenges than without.
Want to serve my country by trying to improve the environment. Want to
bring the best out of me by using my experience and want to take challenging
tasks. God made earth green; it is my desire to keep it Green.
Affiliation: School of Civil Engineering and Surveying, University

Portsmouth, Portland Building, Portland Street, Portsmouth, United Kingdom
PO1 3AH.
Education:
B.E. Kuwait University – Civil Engineer, Main Environmental (2004)
M.S.C Portsmouth University – Civil Engineer and Environmental
Engineer (2009).
Phd Civil Engineer and Environmental Engineer (2015).
Business Address: Lothan Company, Subah Alnasser, Block 7, Home 50,

street 52. Kuwait
Research and Professional Experience:
 Presented the following Topics;

1. Evaluation of a Multi-Stage Strategy for Remediation of Kuwaiti
Oil lakes”
2. Evaluate the standard of Total Petroleum Hydrocarbon (TPH)
For:
 Minister of Oil, Kuwait (Dr. Ali Omair)

 Kuwait Focal Point (KFP)
 Kuwait oil Company (KOC) (R&T with Soil remediation team)
 Kuwait National Petroleum Company (KNPC) (R&T with Soil
remediation team)
 Kuwait Institute of Scientific Research (KISR) (Dr. Samira Asem)
o Assessment of soil remediation strategies for Kuwaiti oil lakes

o Effects of Kuwaiti oil contaminated sand on the engineering
properties of hardened concrete
o Correlation of chemical oxygen demand of highly saline water using
Hg and Hg-Free COD vials
Professional Appointments:
Feb – June 2016: Lecturer, Public Authority of Education and training

Civil Engineering College, Kuwait
 2009-2015: Assistant Researcher, University of Portsmouth, UK

 13-17 April 2014: Lecturer, Public Authority for industry, Kuwait
(Introduction of industrial wastewater)
 15-20 April 2012: Lecturer, Public Authority for industry, Kuwait
(Remediation of contaminated soil and types of hazardous waste
course)
 2004-2009: Civil Engineer in MEW Project, Kuwait
Honors:
 Achievements: Won many awards from different Environmental

organizations, including two Environment Improvement awards from
Government organizations.
 Memorandum of understanding (MOU) is signed with Kuwait
National Patrolmen company (KNPC) for the pilot project for
“Remediation of hydrocarbon contaminated soil.”
 Acknowledgment and appreciation letterhas been givenfrom Kuwait
National Patrolmen company (KNPC) for the pilot project for
“Remediation of hydrocarbon contaminated soil.”
Publications Last 3 Years:
[1] Almutairi, M.S. and Ali, M. Direct detection of saponins in crude

extracts of soapnuts by FTIR, Natural Product Research, 2015, 29 (13),
1271-1275.
[2] Almutairi, M.S. Development and evaluation of a remediation strategy
for the oil lakes of Kuwait. 2016, PhD thesis. University of Portsmouth.
[3] Evaluation and remediation strategy for oil contaminated sand in kuwait
oil lake (2016).
[4] Oral presentation at international conference on contaminated sites,
Bratislava 2016. September 12-13, 2016.
Patents:
[1] Removal of Oil from Sand using Soil Washing Method (US14/945358)
May 13, 2015.
[2] System and Method for Remediation of Oil-Contaminated Sand (Docket
33100.61) 1 August 2016.
Chapter 5
TRITERPENOIDS AND THEIR SAPONINS

FROM ILEX ASPRELLA:
TYPES AND BIOLOGICAL STUDIES
Hui Liu1,2, Fan-Cheng Meng1, Ying Wang2,

Rui-Bing Wang1, Wen-Cai Ye2,* and Qing-Wen Zhang1,†
1
State Key Laboratory of Quality Research in Chinese Medicine,
Institute of Chinese Medical Sciences, University of Macau,
Macao, PR China
2
Institute of Traditional Chinese Medicine and Natural Products,
College of Pharmacy, Jinan University, Guangzhou, PR China
ABSTRACT
The plant Ilex asprella (Hook. et Arn.) Champ. ex Benth.
(Aquifoliaceae) is a deciduous shrub growing in southern China and
Southeast Asia. The roots of I. asprella, known as Gangmei in China,
have been widely used as a Traditional Chinese Medicine (TCM) for the
treatment of headaches, coughs, fever, diarrhea and sore throats. In
addition, its root is key ingredient of ‘Wang-Lao-Ji Herbal Tea’ which
*
Corresponding Author: Tel.: +86 20 8522 0936; fax: +86 20 8522 1559; Email:
chyewc@gmail.com.
†
Corresponding Author: Tel.: +853 8822 4879; fax: +853 20 2884 1358; Email:
qwzhang@umac.mo.
108 Hui Liu, Fan-Cheng Meng, Ying Wang et al.
was formulated in 1828 and has been widely accepted as a healthy

beverage. Previous investigations on the phytochemistry of this plant had
led to the isolation of triterpenoids, flavonoids, phenolic acid and alkaloid
phenolic compounds. The most characteristic constituents of I. asprella
are triterpenoids and their saponins, including some rare sulfur-containing
derivatives. Pharmacological research has identified the important
associations of I. asprella with their anti-cancer, anti-inflammatory, and
anti-virus activities. This chapter will focus on the structural types and
biological studies of saponins of I. asprella, and the characteristics of
these chemical structures will also be briefly discussed.
Keywords: saponin, triterpenoid, Ilex asprella, phytochemistry,

pharmacological properties
INTRODUCTION
Ilex asprella (Hook. et Arn.) Champ. ex Benth. (Aquifoliaceae), a
deciduous shrub about 1-2 meters high, is mainly distributed in Southern
China, such as in Guangdong province and the Guangxi Zhuang autonomous
region. Its roots and leaves are widely used as a Traditional Chinese Medicine
(TCM) for the treatment of headaches, tussis, febris, dysentery, sore throats,
etc. (Cai et al. 2010). The leaves of I. asprella can be used for the treatment of
the inflammation of the upper respiratory tract caused by high fever, enteritis,
and infectious hepatitis (Wang et al. 2009). The roots of I. asprella, called
“Gangmei” in China, have also been an important ingredients for the
preparation of herbal tea including ‘Wang-Lao-Ji Herbal Tea’ formulated in
1828 and beverages that combat viral and bacterial infections for a long time.
Herbal tea became very popular in China and the volume of herbal tea
surpassed Cola in the Chinese market since 2006, which stimulated increasing
interest in phytochemical and biological research on those herbal ingredients.
Phytochemical investigation led to the isolation of saponins, steroids, lignans,
and flavonoids, as well as their glycosides from the roots or leaves of I.
asprella (Wang et al. 2009, Cai et al. 2010). The most characteristic
constituents of I. asprella are triterpenoid saponins, including some rare
sulfur-containing derivatives. Although its roots and leaves were widely used,
only a few phytochemical investigations were conducted before 2010 and
2011 (Cai et al. 2010) respectively. Due to the similarity of saponins
structure，as well as its diversity and complexity, the separation of monomers
of saponins has always been one of the major difficulties encountered by
Triterpenoids and Their Saponins from Ilex asprella 109
researchers investigating the saponins of I. asprella. This chapter will focus on

the types and biological studies on the saponins or their aglycone of I.
asprella, the characteristic of chemical structures will also be briefly
discussed.
CHEMICAL PROPERTIES OF THE TRITERPENOIDS

AND THEIR SAPONINS OF I. ASPRELLA
Triterpenoid saponins are glycosylated triterpenes that are found in

numerous plants and some animals (the most common is the sea cucumber)
(Wina et al. 2005, Lacaille et al. 2010, Riguera et al. 1998). In plants, they
occur in various parts such as the root, tuber, bark, leaves, seeds, and fruits.
Usually, triterpenoid saponins are found principally in dicotyledonous plant
species while steroidal saponins occur in monocotyledons. As a deciduous
dicotyledonous shrub, some triterpenoid saponins were isolated from the roots
and leaves of I. asprella. Saponins are key constituents of I. asprella, and also
regarded as the main active compounds (Yu et al. 2014). Up to now, over 60
triterpenoids and their saponins have been isolated from the leaves and roots of
I. asprella. These triterpenoids from I. asprella can be grouped into two main
types: oleanane type and ursane type. One or more sugar moieties containing
glucose, xylose, arabinose, or glucuronic acid is glycosidically linked to the
sapogenin (aglycone).
Figure 1. Oleanane type triterpene skeleton of I. asprella.

Figure 2. Ursane type triterpene skeleton of I. asprella.
Up to now, triterpenoid saponins bearing aglycones that were connected

with trans-coumaroyl or cis-coumaroyl groups and some rare sulfated
oleanane type saponins have been isolated from the I. asprella leaves and roots
(Peng et al. 2016, Kashiwada et al. 1993). Typically sulfonated saponins are
primarily isolated from marine organisms such as holothurians, seastars and
sponges (Wina et al. 2005). Based on the oxidation degree of the aglycone
moiety, the absolute configuration of C19 and the differences in the substituent
groups, the oleanane-type triterpenoid saponins can be categorized into the
following four types of structures (see Figure 1).
Table 1. Oleanane type triterpenoids and their saponins isolated from

the roots and leaves of I. asprella
No. type R1 R2 R3 R4 Reference

Roots 1 A H H - - He et al. 2012
2 A Glc H - - Lei et al. 2014
3 C H H COOH Glc Lei et al. 2014
4 C H Xyl-2-O- CH3 H Lei et al. 2014
Ac
5 C H Glucuronic CH3 H Lei et al. 2014
acid
6 C H H CH3 H He et al. 2012
7 D Ara-3-O- SO3H Glc - Peng et al. 2016
SO3H
Leaves (1) A H H - - Bai et al. 2011
8 B trans-Coum trans- H - Kashiwada et al.
Coum 1993
9 B trans-Coum cis-Coum H - Kashiwada et al.
1993
10 B cis-Coum trans- H - Kashiwada et al.
Coum 1993
11 B H H H - Kashiwada et al.
1993
12 B H H CH3 - Kashiwada et al.
1993
13 C OH H COOH H Wang et al. 2009
Figure 3. Nortriterpenoid skeleton of I. asprella.

Ursane-type triterpenoid saponins have been mostly isolated from the

roots of I. asprella, and only two of them were isolated from the leaves (see
Figure 2 and Table 2). In comparison with oleanane, structural changes of
ursane-type triterpenoid saponins were more diverse. According to the
oxidation degree of aglycone, the difference in the double bond position and
number, the different absolute configuration of chiral carbon atoms and the
differences in the substituent groups, the ursane-type triterpenoid saponins can
be divided into ten types of structures (see Figure 2).
In addition to triterpenoids, nortriterpenoids have also been isolated from
the roots of I. asprella (see Figure 3 and Table 3).
Table 2. Ursane type triterpenoids and their saponins isolated from

the roots and leaves of I. asprella

Roots 14 E H H CH3 H Cai et al.
2011
15 E H H H CH3 Cai et al.
2011
16 E Xyl Glc CH3 H Cai et al.
2011
17 E Xyl Glc H CH3 Cai et al.
2011
18 E Glucuronic acid Glc H CH3 Wang et al.
methyl ester 2012
19 E Glucuronic acid Glc CH3 H Wang et al.
methyl ester 2012
20 E Xyl H CH3 H Lei et al.
2014
21 E Ara H CH3 H Lei et al.
2014
22 F H H - - Cai et al.
2011
23 F Xyl Glc - - Cai et al.
2011
24 F Glucuronic acid Glc - - Wang et al.
methyl ester 2012
25 F Xyl-2-O- SO3H Glc - - Zhang et al.
2013
26 F Xyl-2-O- SO3H H - - Zhang et al.
2013
27 G Glucuronic acid Glc - - Wang et al.
methyl ester 2012
No. type R1 R2 R3 R4
28 G Xyl -2-O- SO3H Glc - - Zhang et al.
2013
29 H H - - - Huang et al.
2012
30 H Xyl - - - Huang et al.
2012
31 I H H CH3 CH3 Huang et al.
2012
32 I H Glc CH3 CH3 Cai et al.
2011
34 I Xyl H CH3 CH3 Cai et al.
2011
35 I Xyl Glc CH3 CH3 Cai et al.
2011
36 I Glucuronic acid H CH3 CH3 Wang et al.
methyl ester 2012
37 I Glucuronic acid Glc CH3 CH3 Wang et al.
methyl ester -3-O- 2012
SO3Na
38 I Glucuronic acid H CH3 CH3 Wang et al.
methyl ester -3-O- 2012
SO3Na
39 I Glucuronic acid Glc CH3 CH3 Wang et al.
methyl ester 2012
40 I Xyl-3-O- SO3Na Glc CH3 CH3 Zhou et al.
2012
41 I Xyl-3-O- SO3Na H CH3 CH3 Zhou et al.
2012
42 I Xyl(2-1)Glc(2-1)Rha H CH3 CH3 Zhou et al.
2012
43 I Ara Glc CH3 CH3 Zhou et al.
2012
44 I Ara(2-1)Glc H CH3 CH3 Zhou et al.
2012
46 I Glucuronic acid n- H CH3 CH3 Zhao et al.
butyl ester 2013a
47 I Xyl -2-O-Ac H CH3 CH3 Lei et al.
2014
48 I Glucuronic acid H CH3 CH3 Lei et al.
2014
49 I H H CH3 COOH Zhou et al.
2012
50 I H Glc CH3 COOH Zhou et al.
2012
Table 2. (Continued)

51 I Xyl Glc CH2OH CH3 Zhou et al.
2012
52 J H H - - Lei et al.
2014
53 K Ara CH3 - - Wen et al.
2011
54 L H CH3 - - He et al.
2012
55 L Glc H - - Lei et al.
2014
56 M H COOH - - Huang et al.
2012
57 M Ac COOH - - Huang et al.
2012
58 M Ac CH2OH - - Lei et al.
2014
59 N Xyl Glc - - Zhao et al.
2013a
Leaves 60 K H COOH - - Wang et al.
2009
(54) L H CH3 - - Wang et al.
2009
Table 3. Nortriterpenoids isolated from the roots of I. asprella
No. type R1 R2 R3 Reference

Roots 61 O CH3 CH3 CH3 Jiang et al.
2014
62 O H CH3 CH3 Jiang et al.
2014
63 O CH3 CH3 H Jiang et al.
2014
64 O CH3 CH2= - Jiang et al.
2014
BIOLOGICAL EFFECTS OF THE TRITERPENOIDS

AND SAPONINS OF ILEX ASPRELLA
The roots and leaves of I. asprella have been widely used as a Traditional
Chinese Medicine (TCM) to treat headaches, tussis, febris, dysentery, sore
throats for hundreds of years. Furthermore, its roots, also called “Gangmei” in
China, have been key ingredients of herbal tea and beverages that fight viral
and bacterial infections. Modern pharmacological studies have shown that I.
asprella have anti-inflammatory, anti-viral, cytotoxic, antioxidant activities
and other health benefits (Su et al. 2008, Zhao et al. 2013, Eam et al. 2016).
Triterpenoids are the most representative group of phytochemicals, as they
comprise more than 20,000 recognized molecules. Due to their low
hydrophilicity, triterpenes were considered to be inactive for a long period of
time. As the characteristic constituents of Ilex asprella, triterpenes are highly
associated with their broad range of pharmacological effects. The isolated
oleanolic acid [1] (olenane-type), ursolic acid [56] (ursane-type), and other
pentacyclic triterpenoid carboxylic acid are also find in many herbs and fruits,
which have been reported to possess various activities including antioxidant
and anti-tumor activites. The pharmacological activities ascribed to the
compounds found in I. asprella and various parts of I. asprella are detailed
below.
ANTIVIRAL AND ANTIBACTERIAL ACTIVITIES

Many studies have shown the roots and leaves of I. asprella to be a
potential source for antiviral and antibacterial treatments (Su et al. 2008,
Seukep et al. 2016). The water extract of I. asprella has a remarkable ability to
inhibit the cytopathic effect induced by influenza virus, with an EC50 of 88.2
µg raw material/mL (Zhu et al. 2007). In addition, the water extract has a
potent ability to ease lung inflammation of mice infected by aiv influenza virus
H9N2, reduce the lung index, thus decreasing the mortality rate and extending
the survival time of mice infected by H9N2 subtype avian influenza virus (Li
et al. 2012). Another study revealed that the extract of I. asprella at dosages of
500 mg/kg could effectively diminish mortality rates, and ameliorate lung
edema and inflammation, significantly inhibiting the expression of IL-6, TNF-
α and MCP-1, while promoting the expression of IL-10 and IFN-γin serum
(Dai et al. 2014).
Oleanolic acid [1] and ursolic acid [56] are naturally present in many
medicinal herbs. Many studies have shown that oleanolic acid [1] and ursolic
acid can inhibit various bacteria, and the obtained results suggested the
importance of these compounds as antibiotic drugs. Oleanolic acid [1] and
ursolic acid [56] exhibit remarkable antimicrobial activity, and they act against
important human pathogens such as mycobacteria, HIV, and various protozoal
species. Oleanolic acid [1], which was isolated from the roots of I. asprella for
the first time in 2012, can inhibit HIV-1 replication in acutely infected H9
cells with an EC50 value of 1.7 µg/mL, and inhibit H9 cell growth with an
IC50 value of 21.8 µg/mL (Kashiwada et al. 1988). In addition, a MIC of 50
µg/mL was reported when oleanolic acid [1] was used against M. tuberculosis
streptomycin-, isoniazid-, rifampin-, and ethambutol-resistant strains (Jiménez
et al. 2005). Ursolic acid [56], was found to exhibit similar anti-HIV activity,
with an EC50 value of 2.0 µg/mL and IC50 values of 6.5 µg/mL, respectively
(Wolska et al. 2010). Ursolic acid [56] also was reinforced with a minimum
inhibitory concentration (MIC) 90 of 2.0 µg/mL against Streptococcus. mutans
and Streptococcus. sobrinus (Kim et al. 2011). The diversity of the
antibacterial properties exhibited by oleanolic acid [1] and ursolic acid [56]
has also been illustrated against other human bacterial pathogens, such as S.
pneumonia (MIC of 16 µg/mL), methicillin-sensitive and methicillin-resistant
Staphylococcus aureus (MIC value of 8 µg/mL and 64 𝜇g/mL, respectively),
Bacillus subtilis (MIC of 8 µg/mL), B. cereus, Enterococcus faecalis (MIC of
6.25 and 8.00 µg/mL, respectively), E. faecium (MIC of 8 µg/mL), and
Pseudomonas aeruginosa (MIC of 256 µg/mL) (Woldemichael et al. 2003,
Horiuchi et al. 2007, Fontanay et al. 2008, Cunha et al. 2010). Kong et al.
(2013) reported that oleanolic acid [1] and ursolic acid [56] could be used as
potential HCV antiviral agents that can be applied to clinical trials either as
monotherapy drugs or in combination with other HCV antivirals. Further
investigation has shown that both oleanolic acid [1] and ursolic acid [56]
significantly suppressed the replication of HCV genotype 1b replicon and
HCV genotype 2a JFH1 virus by suppressing HCV NS5B RdRp activity as
noncompetitive inhibitors (Kong et al. 2013). Oleanolic acid [1] and ursolic
acid [56] not only influence bacterial growth but also exhibit anti-mutagenic
activity. Oleanolic acid [1] and ursolic acid [56] are also reported to inhibit the
mutagenic activity of benzopyrene towards Salmonella enterica sv.
Typhimurium TA98 (Niikawa et al. 1993). Miyazawa et al. reported the
suppression of the mutagenic activity of the heterocyclic amine (Trp-P-1), a
DNA damaging agent, by ursolic acid [56] (Miyazawa et al. 2005). Pomolic
acid [31] isolated from the roots of I. asprella was identified as an anti-HIV
agent with an EC50 1.4 µg/mL (Kashiwada et al. 1998). In addition to

antiviral activity, pomolic acid was also reported to inhibit the growth of 100%
of the Escherichia coli, E. aerugenes, Klebsiella pneumoniae, Pseudomonas
aeruginosa, Providencia stuartii and Enterobacter cloacae bacterial with
MICs ranged from 32 to 256 µg/mL. Based on the above results, pomolic acid
can be considered as a moderate antibacterial agent, as MIC values ranging
between 10 and 100 µg/mL were obtained on 6/10 (60.0%) tested bacteria
samples. (Seukep et al. 2016). 3-O-β-D-xylo-pyranosyl-3β-hydroxyurs-12, 18
(19)-dien-28-oic acid 28-β-D-glucopyranosyl ester [16], oblonganoside B [23]
and Ziyuglucoside I [44] have shown activity against tobacco mosaic virus
replication with inhibition rates of 33%3, 56.45% and 81.9%, respectively, at a
concentration of 0.2 mg/mL (Wu et al. 2007, Zhang et al. 2007).
Asprellanosides A [40] together with oblonganoside H [35] exhibited anti-
HSV‑1 activity with TIC values of 0.14 and 0.18 mM, respectively (Zhou et
al. 2012). A further study showed that oblonganoside I [51] can moderate anti-
HCV activity with an SI value of 4.41(Zhong et al. 2016). Based on the results
of the structure-activity relationship studies of triterpenes and triterpenoid
glycosides against TMV (Tobacco Mosaic Virus), it was found that the
activities are determined not only by the core structure of triterpenes and
triterpenoid glycosides but also by the substituents on them, which sometimes
can dramatically influence the activity. For the triterpenes of the ursane type,
the anti-TMV activity was increased when a lactone was located at the
positions C-20 and C-28, an arabinosyl glycosylated at C-3 and a glucosyl
linking to C-28. Meanwhile, in the case of the oleanane type, the presence of
an α-hydroxy at C-19, xylosyl at C-3, and glucosyl at C-28 led to a decrease of
the inhibitory activity (Zhang et al. 2007).
CYTOTOXIC AND ANTI-PROLIFERATIVE ACTIVITIES

Numerous studies have demonstrated that compounds isolated from the I.
asprella possessed cytotoxic activity. It has been found that oleanolic acid [1]
was the most effective anti-proliferative agent, as it inhibited HL-60 cell
growth with an IG50 value of 8.9 µM (Wang et al. 2006). Oleanolic acid [1]
was also evaluated for its cytotoxic activities in vitro against lung cancer
(H522), leukemia K562, breast cancer (MCF-7/ADR) and prostate cancer
(DU145) cell lines with IC50 values of 17.85 ± 3.50, 16.15 ± 3.35, 13.82 ±
2.56, and 8.17 ± 1.15 mM, respectively. The in vitro cytotoxic activity assays
were conducted using the classical MTT method (Vimal et al. 2014).
Meanwhile, oleanan-12-ene-3β, 27-diol-28-oic acid [11] showed moderate

growth inhibitory activity selectively toward Leishmania major cells (Zhao et
al. 2013b). In addition, siaresinolic acid [6] inhibited HL-60 cell growth with
IG50 values 29.0 µM (Wang et al. 2006). Kim et al. (2016) reported that
pomolic acid [31] significantly inhibits the invasion of breast cancer cells
possibly through the suppression of CXC chemokine receptor type 4
expression. Ziyu-glucoside I [43] have been tested for their potential
cytotoxicity using an in vitro sulforhodamine B bioassay which showed
significant inhibitory effects against SGC-7901, SMMC-7721, A549, and
DU145 cell lines with IC50 values of 22.08 ± 5.15, 9.27 ± 1.26, 27.80 ± 8.97
and 8.49 ± 0.60 mM, respectively (Zhang et al. 2012). Ziyuglycoside II [53]
has been extensively studied for its pharmacological properties and it has been
found that it can inhibit the proliferation of BGC-823 cells through a possible
mechanism inducing apoptosis but not cell cycle arrest, which was associated
with the regulation of Bax/Bcl-2 expression and activation of the caspase-3
pathway (Zhu et al. 2013a). On the other hand, it could significantly inhibit the
growth of MDA-MB-435 cells by blocking cell cycle progression at G0/G1
and S phase as well as by inducing cell apoptosis (Zhu et al. 2013a, Zhu et al.
2013b). Ursolic acid [56] and its esterified derivatives have also been reported
to show significant cytotoxicities against some tumor cell lines. For instance,
ursolic acid [56] inhibited Akt in human osteosarcoma MG-63 cells via
activation of the ERK1/2 and caspases whereas the pro-survival Bcl-2 and
down-regulated the anti-apoptotic XIAP and survivin (Wu et al. 2016). Young
et al. (Young et al. 1994) reported anti-mutagenic and anti-tumoric activities
of ursolic acid [56] isolated from Eriobotrya japonica Lindl. (Rosaceae).
Ursolic acid [56] also exerted anti-proliferation effects in rat primary VSMCs,
which is associated with the inhibition of miRNA-21 expression and
modulation of the PTEN/PI3K signaling pathway (Jiang et al. 2016). Ursolic
acid [56] inhibited the cell proliferation by inducing apoptosis in HPV-
associated cervical carcinoma cell lines. Some researchers reported that the
apoptotic mechanism of ursolic acid [56] in HeLa cells might be through the
signal pathway “Fas→caspase-8→PARP” and its antiviral effect via
suppression of HPV E6/E7 gene expression which would contribute to growth
inhibition in HeLa cells. Thus, ursolic acid [56] has the potential to serve as an
effective anti-cancer drug for the treatment of HPV-associated cervical
neoplasia (Lee et al. 2003). Meanwhile, 28-O-β-D-glucopyranosyl pomolic
acid [32] inhibited D-GalN-induced cytotoxicity in primary cultured mouse
hepatocytes with an IC50 value of 9.5 µM (Morikawa et al. 2014).
ANTI-INFLAMMATORY AND ANTI-OXIDANT ACTIVITIES

The alcohol extract of roots of I. asprella has a significant inhibitory
effect on rat carrageenan-sex "arthritis,” leukocyte migration and cotton
carrageenan-induced granulomatous inflammation, as well as significant
antagonism to the increased microvascular permeability caused by
inflammatory mediator histamine, but exhibits no obvious antagonism to
increased microvascular permeability caused by inflammatory mediator 5-HT
(Liu et al. 2004). In addition to alcohol extract, the water extract also has been
reported to possess anti-inflammatory activity. The water extract could inhibit
xylene-induced ear edema and cause an increase in capillary permeability in
mice hind paw edema and PGE2 increase induced with carrageen and
granuloma induced with cotton balls in rats (Zhu et al. 2007b). Research has
also shown that the root, stem and root mixture of I. asprella have potent anti-
inflammatory activity and can relieve coughs, while the leaf only exhibits anti-
inflammatory activity (Huang et al. 2012b).
Among the natural products, pentacyclic terpenoids including ursolic acid
[56] and oleanolic acid [1] are considered as promising anti-inflammatory
agents. The major anti-inflammatory effects of these compounds have been
found to be mediated via inactivation of NFкB, STAT3/6, Akt/mTOR
pathways (Dharambir et al. 2016). The protective effect in colitis of ursolic
acid [56] has been reported recently. The mechanism of anti-inflammatory and
antioxidant activities might be through increased the serum levels of IL1β and
TNFα, increased MDA content and decreased SOD activity in the colon
homogenate. Meanwhile, ursolic acid [56] reduced the DSS stimulated high
nuclear level of NFκB p65 in the colon tissues (Liu et al. 2016). Baricevic et
al. (Baricevic et al. 2001) reported that the anti-inflammatory effect of ursolic
acid [56] (ID50 = 0.14 mMoles/cm2) was twice as potent as that of
indomethacin (ID50 = 0.26 mM/cm2), which was used as a reference non-
steroidal anti-inflammatory drug (NSAID). Its anti-inflammatory effects were
evaluated in activated T cells, B cells and macrophages. In order to elucidate
its mechanism of anti-inflammatory activity, the effects of ursolic acid [56] on
ERK, JNK, NF-κB, AP-1 and NF-AT were studied. The in vivo efficacy of
ursolic acid [56] was studied using mouse model of graft-versus-host disease.
Ursolic acid [56] not only inhibited activation, proliferation and cytokine
secretion in T cells, B cells and macrophages, but also inhibited mitogen-
induced up-regulation of activation markers and co-stimulatory molecules in T
and B cells. In addition, it could also inhibit mitogen-induced phosphorylation
of ERK and JNK and suppress the activation of immunoregulatory
transcription factors NF-κB, NF-AT and AP-1 in lymphocytes. The treatment

of cells with ursolic acid [56] prior to allogenic transplantation could
significantly delay the induction of acute graft-versus-host disease in mice and
also significantly reduce the serum levels of pro-inflammatory cytokines IL-6
and IFN-γ. Ursolic acid [56] treatment inhibited T cell activation even when it
was added post-mitogenic stimulation, thus demonstrating its therapeutic
utility as an anti-inflammatory agent (Checker et al. 2012). The anti-
inflammatory activity of siaresinolic acid [6], isolated from the leaves of
Sabicea grisea, was reported by Oliveira et al. (2015). The results indicated
that siaresinolic acid had significant anti-inflammatory effects on carrageenan-
induced pleurisy in mice through mechanisms involving receptors for ATP-
dependent potassium channels (Oliveira et al. 2015). The anti-inflammatory
effects of ziyu-glycoside I [43] were evaluated by nitric oxide (NO) assay. The
inhibitory rate was 24% when the concentration was 40 mg/mL and the IC50
value was 25.77 µg/mL. On the basis of these results, Ziyu-glycoside I showed
potent anti-inflammatory activities (Che et al. 2012). Siaresinolic acid [6]
exhibited protective effects against H2O2‑induced myocardial cell injury. The
SAR studies showed that the methyl ester group at C-28 in triterpene skeleton
decreases its antioxidant activity and the presence of a methoxy group on the
aromatic ring seems to have a negative effect on the antioxidant activity. This
SAR analysis provided a useful information for future synthetic and
pharmacological studies (Li et al. 2014). Bioactivity-guided fractionation of
the ethanol extract of the aerial part of Rubus rigidus var. camerunensis
(Rosaceae) based on free radical-scavenging assay (DPPH assay) afforded
suavissimoside R1 [55], which exhibited a very weak antioxidant effect (about
40%) in DPPH assay (Nguelefack et al. 2011).
ANTI-DIABETIC AND HYPOLIPIDEMIC ACTIVITIES

2β, 3α-Dihydroxyurs-12-en-28-oic acid [52] acts as a natural inhibitor of
glycogen phosphorylase, and exhibits significant inhibition against rabbit
muscle GPa with IC50 values of 1.1µM (Cheng et al. 2008). Zigu-glucoside I
[43] can lower blood glucose levels of db/db mice with reduction rates of
2.6%. The glycated hemoglobin (HbA1c) concentration in db/db mice was
also significantly reduced by treatment of zigu-glucoside I with reduction rates
of 19.4% (Ju et al. 2015). Ursolic acid [56] is a secondary metabolite found in
manyf plants. Activity-directed fractionation of the ethanol extract of aerial
partsof Artemisia roxburghiana has led to the isolation of ursolic acid [56],
which is known to inhibit protein tyrosine phosphatase (PTP) 1B with IC50

values of 3.21 ± 0.02 µM (Shah et al. 2016). As competitive inhibitors of
PTP1B, ursolic acid [56] also inhibits T-cell PTP and src homology
phosphatase-2 but not leucocyte antigen-related phosphatase or PTPs α and ε,
which are all possibly involved in the control of insulin level. Ursolic acid [56]
is about twice as potent as corosolic acid which has a similar structure and
often co-exists with ursolic acid [56] in the plant (Zhang et al. 2006, Na et al.
2005). After streptozotocin -induced diabetic mice being administered of a low
dose of ursolic acid [56] (0.2% of the diet) for 11 weeks, it was found that
ursolic acid [56] was able to reduce by 53% the inherent lesions that occurred
in the endothelium with diabetes. In all of the above cases thus compound
displayed a protective but not necessarily rehabilitative effect. Ursolic acid
[56] displayed a dose-dependent inhibition of H2O2-accelerated chemotaxis in
response to MCP-1 with an IC50 of 0.4 µM which was 2.7-fold more potent
than resveratrol in the same assay (Ullevig et al. 2011). Ursolic acid [56] at
dosages of 4-20 µM appears to facilitate rosiglitazone-induced glucose uptake
into insulin resistance fat cells in vitro, although combination treatment was
associated with less adipocyte differentiation than rosiglitazone alone (Li et al.
2012). It promoted lipolysis in vitro by activating Protein Kinase A, and
downstream of Hormone Sensitive Lipase and Perilipin A activity. It was also
found that upregulation of Adipose Tissue Lipase in adipocytes is independent
to PKA (Li et al. 2010).
Song et al. found that oleanolic acid [1] exhibited a glucosidase inhibitory
activity with an inhibition ratio of 2.92% (Song et al. 2010).
HEMOSTATIC AND ANTITHROMBOTIC ACTIVITIES

Bioassay-guided separation of the extract disclosed zigu-glucoside I [43]
as a new hemostatic molecule, which showed the strongest hemostatic activity
with a Goat anti-human α2-plasmin inhibitor ELISA kit (Sun et al. 2012). The
methanol extract of I. pubescens roots prolonged bleeding time by three fold,
and inhibited the generation of malondialdehyde released during platelet
aggregation induced by thrombin, which might be attributed to the presence of
Ilexoside A [59] and Ilexoside B [34] (Han et al. 1987).
OTHER DIVERSE ACTIVITIES

In addition to the above mentioned activities, many other activities were
also reported. The gangmei compound granules had significant antitussive
effect induced by inhalation of ammonia water aerosol in mice which
suggested that gangmei compound granules had an antitussive effect that was
induced by pharyngitis and laryngitis in humansman (Zhoug et al. 2005).
Triterpenoids and the saponins isolated from I. asprella have also been
reported to exhibit other diverse activities. Ursolic acid [56] reduced the
muscle atrophy and stimulated muscle hypertrophy by enhancing skeletal
muscle insulin/IGF-1 signaling and inhibiting atrophy associated skeletal
muscle mRNA expression (Kunkel et al. 2011). Another report disclosed that
ursolic acid [56] directly promoted protein accretion in cultured myotubes but
did not modulate myoblast proliferation and concluded that the anabolic effect
is direct on the muscle cell. ((Figueiredo et al. 2012). A testosterone-induced
benign prostatic hyperplasia rat model using ursolic acid [56] oral
administration (5 mg/kg) revealed a significant suppression in prostate weight
and testosterone and DHT levels in both the serum and prostate, which suggest
that ursolic acid [56] effectively inhibit the development of BPH and it may be
a potential drug in BPH treatment (Shin et al. 2012). Acetylursolic acid [57]
exhibited moderately anti-plasmodial activity with IC50 values of 12.09 ± 0.08
µM (Su et al. 2016). Ilexsaponin B [17] exhibited moderate xanthine oxidase
inhibitory activity in the test with an IC50 value of 26.01µM (Zhou et al.
2013). Suavissimoside R1 [55] exhibited a protective effect on dopaminergic
neurons against MPP+ toxicity (Yu et al. 2008).
CONCLUSION
For many years, I. asprella has been widely used as a Traditional Chinese
Medicine (TCM) for the treatment of many diseases. Its roots and leaves were
also important ingredients for the preparation of herbal tea and beverages in
Chinese folk medicine. Before 2010, there were few phytochemical
investigations and biological studies of saponins of I. asprella. There are only
64 saponin monomers that are known at present. Due to the similarity of
saponins structure，as well as its diversity and complexity, the separation of
monomers of saponins has always been one of the major difficulties that has
hindered research on the saponins from I. asprella. With the rapid
development of separation technology and growing interest in natural, the

saponin monomers from I. asprella will be investigated more extensively.
In addition, the water or alcohol extract displayed stronger antiviral and
antibacterial activities. Many monomer saponins also exhibit a diverse range
of antiviral, antibacterial, cytotoxic, anti-inflammatory, antioxidant, anti-
diabetic, hypolipidemic, hemostatic and antithrombotic activities. Meanwhile,
the pharmacological activities of most the saponins, especially the sulfonated
saponins, from I. asprella haven’t been screened. Therefore, the
phytochemical and pharmacological properties of I. asprella need to be further
investigated. We hope that more structurally unique saponins with diverse
pharmacological activities will continue to be discovered from I. asprella.
ACKNOWLEDGMENTS
This work was supported by grants from Macao Science and Technology
Development Fund (FDCT/042/2014/A1) and Ministry of Science and
Technology of China (No. 2013DFM30080).
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INDEX
aflatoxin, 129
A agar, 12, 96
Agave sisalana, 11, 31, 72, 87
A. minutiflorum, 11
Agave spp., 11
Acacia auriculoformis, 12
age, 21, 104
accessibility, 88
aglycone, vii, 1, 3, 7, 10, 42, 43, 64, 92,
acetylcholine, 43
109, 110, 112
Achyranthes aspera, 13, 29
agonist, 24
acid, x, 7, 11, 12, 13, 19, 29, 35, 42, 43, 57,
agriculture, 87
64, 66, 71, 78, 81, 85, 108, 109, 111,
AIDS, 24, 125
112, 113, 115, 116, 117, 119, 120, 121,
alfalfa, 65
122, 123, 124, 125, 126, 127, 128, 129
algae, 65
acidic, 18, 26, 47, 70, 75, 81
alkaloids, 13
active compound, 68, 109
Allium, 11, 22, 27, 68, 86
acute lung injury, 16
amine, 116, 126
acute respiratory distress syndrome, 124
ammonia, 122
acylation, 13
amphiphilic, vii, ix, 1, 3, 57, 63, 64, 68, 92
additives, 5, 78
amyloid beta, 23, 30
adenocarcinoma, 12
Anagallis arvensis, 11, 13, 21
adipocyte, 40, 41, 42, 44, 46, 47, 48, 49, 51,
analgesic, 17
121
androgen, 50
adipogenesis, 38, 39, 41, 44, 46, 47, 49, 50
Anemarrhean asphodeloides, 20
adiponectin, 42
angiogenesis, 49
adipose tissue, 38, 39, 47
antagonism, 119
adiposity, 46, 52
antibacterial, viii, 2, 11, 12, 115, 116, 123,
adjustment, 79
124, 127, 128
adjuvants, 5, 6, 8, 21, 22, 29, 30, 31, 32, 33
antibiotic, 116
ADR, 117
antibody, 8, 22, 24, 25, 32, 34
adsorption, 76, 78, 79
anti-cancer, x, 53, 56, 108, 118
adults, 32, 49, 50
anti-diabetic, 120, 123
Aesculus hippocastanum, 17, 67, 68, 69, 73
132 Index
antifungal, viii, 2, 5, 10, 11, 22, 24, 27, 29,

30, 32, 86
B
antigen, 5, 7, 8, 9, 24, 27, 31, 33, 34, 121
Bacillus subtilis, 11, 116
antigen-presenting cells (APCs), 7
Bacopa monnieri, 18, 87
anti-inflammatory, viii, x, 2, 5, 15, 16, 17,
bacteria, 6, 12, 43, 46, 57, 94, 116, 127
23, 25, 35, 56, 108, 115, 119, 123, 124,
bacterial infection, 108, 115
125, 126, 127, 130
bacterial pathogens, 116
anti-inflammatory agents, 119
Balanites aegyptiaca, 11
antimicrobial, 5, 9, 11, 12, 22, 24, 25, 26,
beneficial effect, viii, 37, 38, 42
28, 30, 31, 86, 87, 116, 124, 125
benefits, 29, 93, 115
anti-obesity, viii, 37, 38, 40, 45, 48
benign prostatic hyperplasia, 122, 127
antioxidant, viii, 2, 15, 17, 18, 19, 28, 31,
benzene, 77
50, 51, 87, 88, 115, 119, 123, 124, 126
beverages, 108, 115, 122
antioxidative activity, 43, 57
bidesmosidic, 3, 64
anti-proliferative, 117
bile acids, 14, 86
antithrombotic, 121, 123, 124
bioaccumulation, 72
antitumor, 5
bioassay, 118
antiviral, viii, 2, 5, 13, 14, 21, 26, 32, 115,
biochemistry, 124, 125
116, 118, 123, 125, 129
biodegradability, 82, 94, 102
antiviral agents, 116
biodiesel, 85
apoptosis, viii, 2, 16, 19, 20, 27, 118, 128,
biodiversity, 126
130
biological activities, vii, viii, 2, 15, 24, 31,
appetite, 41, 42
34, 43, 46
aquifers, 83
biological activity, 32
armed forces, 94
biomass, 82
arsenic, 75, 79, 84, 85, 86, 88
biosurfactant, ix, 71, 75, 79, 81, 83, 85, 87,
arteriosclerosis, 38
88, 89, 91, 92, 94, 95, 103
artery, 19
biosynthesis, 42, 47
arthritis, 16, 23, 119
bleeding time, 121
Asia, 14, 66
blood, 39, 96, 120
asparagus, viii, 2, 65
Blupleurum kaoi, 14
aspartate, 25
body fat, 38, 41
Astragalus membranaceus, 16
body weight, 38
Astragalus verrucosus, 12, 30
bone, 46
astrocytes, 28, 58
brain, 54
atherosclerosis, 128
Brazil, 1, 8
ATP, 26, 42, 120
breast cancer, 51, 117
atrophy, 33, 122, 125
breast carcinoma, 130
avian, 9, 34, 115
Bupleurum spp, 14
avian influenza, 9, 34, 115
awareness, 93, 94
axons, 17, 19 C
Ca2+, 18, 19, 20, 43

cadmium, 83, 85, 87, 88, 89
Index 133
caffeine, 38 children, 38
calcium, viii, 2, 20, 54, 57, 59 Chile, 66
Camellia sinensis, 16 China, ix, 13, 16, 20, 43, 66, 107, 108, 115,
Camellia sp., 11 123, 124
cancer, viii, 6, 37, 38, 45, 50, 51, 53, 56, 57, chinese medicine, 48
59, 60, 118 cholera, 9
cancer cells, 6, 53, 118 cholesterol, 7, 10, 14, 26, 28, 39, 42, 44, 86
cancer stem cells, 53 choline, 81
capillary, 70, 119 chromatographic methods, viii, 2, 4
Capsicum annuum, 12 chromatographic technique, 4
carbohydrates, 13 chromatography, viii, 2, 4, 66
carbon, vii, 1, 43, 74, 81, 112 chronic venous insufficiency, 84
carbon atoms, 112 circadian rhythm, 54
carboxyl, 77 circulation, 16
carboxylic acid, 115 cities, 82
carboxylic groups, 70 classification, 33, 93
carbuncle, 16 climates, 65
carcinoma, 118, 130 clinical application, 48
cardiovascular disease, viii, 37, 44, 51 clinical trials, 6, 8, 116
cardiovascular disorders, 45 CMC, ix, 64, 71, 72, 74, 100, 102
caspases, 27, 118 colitis, 119
cell culture, 22 colon, 119
cell cycle, 118, 130 Columbrina retusa, 10
cell death, 20, 26 commercial, ix, 6, 8, 34, 63, 65, 70, 71, 72,
cell fate, 41 74, 75, 76, 77, 82, 95
cell line, 12, 15, 117 competition, 74, 77
cellular energy, 42 complexity, 108, 122
central nervous system, 39, 48 complications, 38
cerebrovascular disease, 20 composition, viii, 2, 8, 9, 13, 14, 28, 29, 38,
CGAs, 81 67, 68, 71, 76
challenges, 104 compounds, vii, viii, 1, 2, 4, 5, 8, 9, 11, 13,
cheese, 57 14, 21, 30, 37, 38, 79, 81, 93, 94, 108,
chemical, vii, viii, ix, x, 2, 3, 4, 5, 9, 10, 12, 115, 116, 117, 119, 124, 127
13, 14, 38, 42, 48, 50, 63, 64, 73, 76, 78, conductivity, 97
82, 84, 92, 94, 105, 108, 109, 126, 127, conference, 106
128 configuration, 110, 112
chemical characteristics, 4 connective tissue, 39
chemical properties, 5 connectivity, 21
chemical structures, vii, x, 108, 109 consensus, 46
chemokine receptor, 118 constituents, x, 46, 82, 94, 108, 109, 115,
chemokines, 16 123, 124, 126, 128
chemoprevention, 50 contact time, 93
chemotaxis, 9, 121 containers, 96
Chenopodium quinoa, 8, 9, 10, 24, 32, 33, contaminated sites, 77, 102, 106
34, 65
134 Index
contaminated soil(s), vii, ix, 70, 73, 75, 76, DESs, 81

78, 83, 85, 86, 87, 88, 91, 94, 95, 96, 99, destruction, vii, ix, 91
106 detection, 47, 97, 106
contamination, 77, 94, 99, 100, 102 detergents, 87
controlled trials, 8 developed countries, 15
convergence, 61 diabetes, viii, 37, 38, 51, 121
COOH, 64, 92, 111, 113, 114 diarrhea, x, 8, 23, 107
coordination, 39 diet, 38, 48, 49, 51, 121
copper, 83 diffusion, 12, 74
Cordyline fructiosa, 12 digestion, 43, 58
coronary heart disease, 20 disease model, 21
correlation, 34 diseases, 14, 16, 19, 38, 45, 122
cortex, 26 disorder, 20
cortical neurons, 30 dispersion, 67
cosmetics, ix, 5, 63, 67, 93 distilled water, 96, 97
cost, 73, 78, 94, 95, 100 distribution, vii, 1, 2, 32, 41, 72, 77, 84, 85,
cotton, 119 87
coxsackievirus, 14, 32 diversity, viii, 2, 5, 50, 108, 116, 122
crude oil, 88, 94, 101 DNA, 34, 40, 116
cultivation, 67, 87 donors, 81
culture, 12, 28, 83 dopamine, 26
culture medium, 12 dopaminergic, 20, 122, 129
CXC, 118, 125 dorsal horn, 18
cyclophosphamide, 43, 51, 56 dosage, 74, 79, 82
cytokines, 6, 16, 17, 41, 120 dose-response relationship, 17
cytomegalovirus, 7 down-regulation, 16
cytoskeleton, 48 drugs, 18, 38, 116, 126
cytotoxic, 5, 7, 12, 25, 115, 117, 123, 125, drying, 14, 38, 97
128 Drypete inaequalis, 12
cytotoxicity, 13, 28, 32, 34, 118, 129 dyslipidemia, 46
D E
D. laciniata, 12 E. coli, 12
decomposition, 75 economic losses, viii, 37
deficiency, 81 economics, 86
degradation, 42, 82 EDDS, 78, 83
dementia, 20, 30 edema, 115, 119
depolarization, 18 effluent, 79
depression, 54 electric conductivity, 96
deprivation, 20, 35 electric field, 58
depth, 96 electrical conductivity, 99
derivatives, x, 12, 39, 86, 108, 118, 124, electrolyte, 93
127, 128 electron, 81
desorption, 78, 83 electrophoresis, 54, 58
Index 135
endocrine, 39, 47
endocrine system, 39
F
endocrinology, 49
falciparum malaria, 32
endothelial cells, 43
families, vii, 1, 65
endothelium, 121
family members, 40
energy, 39, 41, 42, 47, 48, 51, 52
fasting, 42
energy expenditure, 39, 52
fat, 31, 39, 47, 48, 51, 121
engineering, 105
fatty acids, 38, 42
Enhanced Oil Recovery, 85
fermentation, 57, 83
Entada phaseoloides, 16, 35
fertility, 67
enteritis, 108
fever, x, 107, 108
enterovirus, 14, 32
fibrosis, 47
environment, 74, 87, 94, 104
films, 80
environmental aspects, 46
filtration, 66
environmental conditions, 70, 94
fires, 99
environmental factors, vii, 2
flavonoids, x, 13, 53, 108
environmental impact, 73, 94
flocculation, 74, 79
Environmental Protection Agency (EPA),
flowers, 65, 67
77, 88, 96
foams, 3, 69, 80, 81, 82
enzyme-linked immunosorbent assay
food, viii, ix, 30, 37, 39, 44, 63, 66, 126,
(ELISA), 4, 121
128
enzyme(s), 4, 19, 42, 59, 127
food additive, 66
epidemiology, 50
food intake, 39
erythrocytes, 10
formation, 10, 15, 69, 70
ESI, 5
France, 52
ESR, 48
fructose, 81
ester, 26, 64, 66, 112, 113, 117, 120, 125
fruits, 24, 67, 68, 95, 109, 115
ethanol, 95, 97, 120, 126
FTIR, 95, 97, 106
ethylene, 81
functional food, 38, 43, 45
ethylene glycol, 81
funding, 88, 103
Europe, 43, 67
fungi, 11, 22, 57
European Commission, 87
fungus, 10
evidence, 16, 20, 49
evolution, 22
excitotoxicity, 18, 26 G
excretion, 14
exercise, 38, 39, 42 GAO, 88
expectorant, 5 gel, 54, 58
extraction, 4, 22, 65, 66, 68, 73, 74, 75, 76, gene expression, 40, 41, 44, 50, 51, 118
84, 85, 89, 95, 97 genes, 15, 40
extracts, 11, 12, 22, 25, 30, 45, 56, 66, 68, genotype, 116
69, 82, 83, 97, 106, 127 Germany, 95
ginseng, viii, 8, 11, 14, 17, 18, 20, 21, 26,
32, 37, 38, 42, 45, 46, 47, 48, 49, 50, 51,
53, 56, 57, 59
136 Index
Gleditsia sinensis, 16, 23 Hedera colchica, 10, 29

gluconeogenesis, 42 Hedera helix, 14, 25
glucose, 20, 31, 35, 42, 44, 49, 64, 109, 120, Hedyotis nudicaulis, 11, 27
129 height, 69
glucoside, 118, 120, 121 Helicobacter pylori, 31
GLUT4, 44 hemoglobin, 120
glutamate, 18, 21, 25 hemostatic, 121, 123, 127
glutathione, 18, 19 hepatitis, 7, 9, 13, 14, 27, 28, 33, 34, 35
glycerol, 81 hepatocytes, 118
Glycine max, 8, 9, 65 Herniaria hirsuta, 15, 24
glycogen, 120, 123 herpes, 7, 8, 13, 14, 29
glycol, 81 herpes simplex, 7, 13
glycoproteins, 13 herpes simplex virus type 1, 7
glycoside, 67, 120, 127, 129 herpes virus, 8, 13, 14, 29
God, 104 hexane, 12
gonorrhea, 12 high fat, 49
graduate students, 55 hipocholesterolemic, 5
grants, 123 hippocampus, 20, 26, 58, 60
granules, 122, 129 histamine, 119
grass, 77, 89 human immunodeficiency virus (HIV), 7, 8,
greenhouse, 68 13, 14, 24, 29, 34, 116, 125
growth, 10, 12, 41, 47, 56, 116, 117, 128, HIV-1, 7, 24, 29, 34, 116
130 homeostasis, 14, 42
growth factor, 41, 47, 56 homocysteine, 26
growth hormone, 41 hormones, 39, 41, 45
Guangdong, 108, 128, 130 host, 119
Guangzhou, 107 HPV, 118
Guinea, 65 human, 6, 7, 8, 14, 17, 20, 32, 34, 43, 45,
Gynostemma pentaphyllum, 15 46, 50, 51, 54, 58, 59, 60, 87, 116, 118,
121, 125, 128, 130
human body, 45
H human health, 87
humidity, 67
haemolytic, 5, 9, 10, 12, 13, 21, 25, 28, 30,
hydrocarbons, 96, 102
31, 32, 33, 34
hydrogen, 81
hardness, 72
hydrolysis, 43
hazardous waste, 77, 106
hydrophilicity, 115
hazards, 87
hydrophobicity, 94
HBD, 81
hydroxide, 79
HBV, 8, 13
hydroxyl, 43, 48, 66
health, 21, 27, 29, 38, 43, 47, 115
hydroxyl groups, 66
health care, 27
hygiene, 130
health care system, 27
hyperlipidemia, 38
health effects, 47
hyperplasia, 39
heavy metals, ix, 64, 73, 74, 75, 76, 77, 78,
hypertension, 38
79, 80, 82, 83, 84, 85, 87, 89
Index 137
hypertrophy, 39, 122 inflammatory responses, 16, 35

hypolipidemic, 120, 123 influenza, 8, 14, 23, 25, 115, 124, 126
hypothalamus, 42 influenza vaccine, 23
hypothesis, 95 influenza virus, 8, 14, 115, 124, 126
hypoxia, 19, 28 Inga laurina, 8, 9, 23
ingredients, 108, 115, 122
inhibition, viii, 2, 12, 16, 19, 21, 41, 44,
I 117, 118, 120, 121, 125, 127
inhibitor, 120, 121, 123
identification, 4, 129
injury, 17, 19, 20, 31, 120, 126
IFN, 115, 120
insulin, 38, 41, 46, 49, 50, 51, 121, 122,
IL-8, 16
126, 129
Ilex asprella, v, ix, 107, 108, 115, 123, 124,
insulin resistance, 46, 121, 126
125, 126, 127, 128, 129
insulin signaling, 49
Ilex paraguariensis, 15, 31
integration, 47
Ilex spp., 13
interferon gamma, 6
image, 57
intestine, 15
immune disorders, 38
intracellular calcium, 17
immune response, viii, 2, 5, 6, 9, 15, 22, 23,
intramuscular injection, 18
25, 27, 29, 34, 35
ion channels, 48
immune system, 5, 6
ionization, 5
immunity, 5, 6, 29, 56
ions, 75, 81, 99
immunization, 24, 31, 32
Iowa, 52, 53, 54, 55
immunoadjuvant, 5, 6, 8, 27, 33
Iran, 29, 87
immunogenicity, 5, 32, 33, 34
Iraq, 94
immunomodulatory, viii, 2, 8
irradiation, vii, 2
immunoreactivity, 18
IRS, 41
immunosuppression, 15
ischemia, 19, 20, 27, 35
improvements, 9
ischemia reperfusion injury, 19
in vitro, viii, 2, 13, 18, 21, 26, 31, 32, 41,
ISCOMs, 7, 22
46, 51, 53, 59, 60, 83, 117, 121
isolation, x, 9, 108, 120
in vivo, viii, 2, 18, 26, 28, 31, 32, 41, 53,
isomers, 43
56, 59, 60, 119, 126
isoniazid, 116
India, 65
Israel, 22
individuals, viii, 37
induction, 24, 41, 120, 130
industries, 5 J
industry, 76, 84, 106
INF, 6 Japan, 43, 66, 74
infection, 14 Joazeiro, 11, 31, 72, 87
infectious agents, 6
infectious hepatitis, 108
inflammasome, 29 K
inflammation, viii, 2, 14, 15, 16, 19, 46,
K+, 19, 43
108, 115, 119
kaempferol, 56, 57
inflammatory disease, 16
138 Index
KBr, 97
Kenya, 32
M
Korea, 16, 37, 43, 45, 54, 56, 99
macrophages, 119
Kuwait, vii, ix, 91, 93, 94, 96, 99, 100, 103,
Maesa lanceolata, 10, 13, 21, 31, 68, 88
105, 106
majority, 70
malaria, 8
L management, 18, 76, 82
mass, viii, ix, 2, 4, 39, 48, 54, 58, 80, 91, 98
lactate dehydrogenase, 19 mass spectrometry, viii, 2, 4, 48, 58
Lactobacillus, 12, 56, 57, 59 materials, viii, 37, 44, 45, 129
lakes, 93, 94, 103, 105, 106 MCP, 115, 121
laryngitis, 122 MCP-1, 115, 121
LC-MS, 4 media, 96
leaching, 76 mediation, 16
leakage, 19 medicine, 15, 16, 33, 122, 126, 130
learning, 20, 30 melanoma, 22
leishmaniasis, 26, 27, 29, 33 mellitus, 38
leptin, 41, 42 melting, 81
lesions, 16, 121 memory, 16, 20, 21, 23, 30, 33
leucocyte, 121 memory B cells, 16
leukemia, 117 memory formation, 21
life expectancy, viii, 37, 44 messengers, 19
ligand, 73 metabolic disorder, viii, 37, 38, 44
light, vii, 2, 67 metabolism, 15, 39, 41, 42, 46, 47, 48, 125
lignans, 108 metabolites, vii, ix, 1, 2, 63, 64, 125
lipid accumulation, 38, 40, 41, 44 metabolized, 43
lipid metabolism, vii, 39, 40, 42, 45, 49 metal extraction, 77
lipids, 42 metal ion, 93
lipodystrophy, 47 metals, ix, 64, 72, 73, 74, 75, 76, 77, 79, 84,
lipolysis, 44, 51, 121, 126 100
liposomes, 7 methanol, 12, 121, 127
liquid chromatography, viii, 2, 4, 48 methodology, 46
liquids, 82, 85 methylene chloride, 124
liver, 14, 42 Mexico, 66
liver disease, 14 mice, 6, 8, 9, 14, 15, 18, 20, 21, 22, 23, 24,
LTD, 96 25, 28, 29, 32, 34, 35, 43, 47, 49, 51,
Luffa operculata, 12, 31 115, 119, 120, 122, 124, 125, 126, 128,
lumen, 15 129
lung cancer, 44, 49, 117 microbiota, 50
Luo, 19, 20, 27, 28, 125 microorganism(s), 12, 43, 82
lymphocytes, 28, 120 microRNA, 125
lymphoid, 41 microscope, 17
migration, 15, 44, 49, 74, 119
Ministry of Education, 45, 60
mitochondria, 19, 20, 130
Index 139
mitogen, 17, 41, 119 neurotoxicity, 25, 28, 30

mixing, ix, 64, 81, 99 neurotransmitter(s), viii, 2, 19
moderate activity, 12 neurotrophic factors, viii, 2, 19
mold, 11 neutral, 72, 73, 93
molecular structure, 73 next generation, 22
molecules, vii, 1, 5, 39, 40, 41, 45, 72, 74, nickel, 83
77, 115, 119 Nile, 28
monodesmosidic, 3, 10, 64 nitric oxide, 19, 120
monomers, 71, 108, 122 nitrite, 20
Moon, 56, 59 NMR, 29
morphine, 18, 30, 35 nonionic surfactants, 70
morphology, 81 non-polar, 69, 92
mortality rate, 115 nortriterpenoids, 112, 114, 125
mosaic, 13, 117, 129 Nrf2, 16, 23, 56
mRNA, 122, 125 nucleus, vii, 1, 40
mucosa, 5 nutraceutical, 38
mucus, 17 nutrient, vii, 2
multipotent, 60 nutrition, 39, 126
municipal solid waste (MSW), 76, 85
muscle atrophy, 122
muscle mass, 125 O
musculoskeletal, 18
obesity, viii, 37, 38, 39, 40, 41, 44, 45, 46,
mycobacteria, 116
47, 48, 49, 50, 51, 52
myelin, 17
occlusion, 19
oil, vii, ix, 57, 64, 65, 66, 70, 78, 88, 89, 91,
N 93, 94, 95, 96, 97, 99, 100, 101, 102,
103, 105, 106
Na+, 19, 43 oleanane type triterpenoids, 111
National Academy of Sciences, 51 oligosaccharide, 7
National Institutes of Health, 20 operations, 95
native species, 6, 8 optimization, 98
natural compound, 38, 81, 125 organic compounds, 77
natural resources, 45 organic matter, 74, 76
nerve, 17 organs, 42
nerve conduction velocity, 17 overproduction, 18
nervous system, 19 oxidation, viii, 2, 31, 42, 110, 112
neural network, viii, 2, 19 oxidative stress, 15, 35, 43, 51
neurodegeneration, 38 oxygen, 20, 35, 105
neurodegenerative diseases, viii, 2, 19
neurodegenerative disorders, 20
neuronal apoptosis, 21 P
neurons, 15, 18, 19, 20, 21, 34, 35, 53, 122,
P. ginseng, 8
129
P. notoginseng, 8, 10, 15, 19, 20
neuropathic pain, 17
p53, 56
neuroprotection, 19, 20, 21, 31
140 Index
pain, 15, 17, 18 polar, 65, 92

palm oil, 85 pollutant, 70, 78, 80, 82
Panax spp, 8 pollutants, ix, 63, 64, 69, 70, 73, 76, 77, 78,
pathogens, 6, 11, 116 80, 82
pathophysiological, 38 pollution, 94
pathophysiology, 47 polyacrylamide, 54
pathway(s), 16, 21, 31, 40, 41, 42, 44, 48, polychlorinated biphenyl, 89
50, 118, 119, 128, 130 polycyclic aromatic hydrocarbon (PAHs),
PCBs, ix, 64, 78, 89 ix, 64, 77, 78, 87, 89
peptide, 23, 30, 58 polymer, 57, 58, 60
permeability, 119 polysaccharides, 38
pests, 3 population, 94
petroleum, 83, 94, 96, 105 porous media, 80
pH, 17, 70, 72, 73, 74, 75, 76, 78, 79, 80, potassium, 120
86, 94, 97, 99 Potentilla anserine, 13, 35
phagocytosis, 9 precipitation, 76, 78, 79
pharmaceutical, ix, 5, 13, 63, 128 pregnancy, 54, 58
pharmacological treatment, 47 preparation, 31, 38, 56, 86, 96, 97, 108, 122
pharmacology, 14, 46, 48 preservation, 38
pharyngitis, 122 preservative, 89
phenolic compounds, x, 38, 108 prevention, 13, 45, 47
phenotypes, 127 progenitor cell, 20, 52, 57, 58, 59, 60
phospholipids, 7 pro-inflammatory, 17, 120
phosphorylation, viii, 2, 19, 40, 41, 119, 129 project, 61, 106
physical activity, 39 proliferation, 20, 57, 59, 118, 119, 122, 124,
physicochemical properties, ix, 63 125
Phytolacca octandra, 11 prostate cancer, 117
Phytolacca tetramera, 10, 24 protection, 3, 6, 25, 29, 30
phytomedicine, 48 proteins, 17, 40, 48, 53, 54
phytoremediation, 80, 82, 89 proteome, 54
PI3K, 41, 118, 125 Pseudomonas aeruginosa, 12, 116
placebo, 20, 28 psychiatric disorder, 53
plant growth, 87 psychosocial factors, 39
plants, 3, 4, 9, 30, 35, 38, 56, 65, 67, 71, 72, PTEN, 118, 125
82, 88, 92, 109, 120, 125 Pulsatilla chinensis, 21
plasma membrane, 17, 21 purification, 9, 66, 95
plasticity, 20, 21 purity, 68, 71, 95
platelet aggregation, 121
platinum, 99
Platycodon grandiflorum, 13, 29, 32, 34 Q
Platycodon grandiflorus, 8, 9
QS-21, 6, 7, 8, 22, 24, 25, 27, 29
pleurisy, 120
quality control, 25
PM, 23, 24, 25, 31, 33, 35
quality of life, 21
pneumonia, 116
quantification, viii, 2, 5, 22, 84
Poland, 63, 75
quaternary ammonium, 81
Index 141
quercetin, 56, 57
Quil-A®, 6, 8
S
Quillaic acid, 7, 66
saccharide side chains, 3, 10
Quillaja brasiliensis, 8, 23, 24, 26, 31, 35
safety, 8, 28, 32, 38, 84, 102
Quillaja saponaria, 6, 11, 25, 26, 27, 29,
saline water, 105
68, 69, 87, 92, 95
salinity, 70
Salmonella, 12, 116, 126, 129
R salt concentration, 72
salts, 73
rabies virus, 8 saponin(s), v, vii, ix, x, 1, 2, 3, 4, 5, 6, 7, 8,
Rapanea melanophloeos, 10, 30 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,
rare earth elements, 84 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,
raw materials, 95 31, 32, 33, 34, 35, 38, 42, 47, 49, 63, 64,
reactive oxygen, 18, 19 65, 66, 67, 68, 69, 70, 71, 72, 73, 75, 76,
receptor, 17, 18, 19, 25, 39, 41, 43, 50, 51, 80, 82, 83, 84, 85, 86, 87, 88, 89, 92, 93,
120, 129 94, 95, 97, 106, 107, 108, 109, 110, 111,
recovery, vii, ix, 58, 64, 78, 79, 84, 85 112, 115, 122, 123, 124, 125, 126, 128,
recycling, 79 129
regenerate, 19 SAR, 120
regeneration, viii, 2, 19 SDS, 54, 81, 86
remediation, vii, ix, 63, 64, 67, 68, 73, 74, secrete, 41
75, 76, 77, 78, 80, 82, 85, 87, 88, 92, 93, secretion, 7, 50, 119
94, 102, 103, 105, 106 seed, viii, 2, 65, 67, 77, 87
replication, 34, 116, 128 selectivity, 73, 82, 124
repulsion, 70 sensor, 48
researchers, 75, 109, 118 sepsis, 15, 28
residue(s), vii, 1, 7, 92, 95, 99, 100, 101 serine, 42
resistance, 44, 49 serotonin, 16, 43, 54, 58
resources, 61 serum, 115, 119, 122
respiratory syncytial virus, 14, 32 sex, 41, 119
response, 5, 8, 15, 17, 22, 28, 29, 33, 41, 45, sex hormones, 41
121 SF, 80
restoration, 17 side chain, 3, 10
resveratrol, 121 signaling pathway, 16, 40, 44, 48, 118
rhamnolipid, 77 signals, 41
risk, 24, 38, 45, 73 sinusitis, 12
room temperature, 72, 74 skeletal muscle, 122, 125
root(s), ix, 13, 14, 17, 28, 29, 32, 34, 38, 51, skeleton, 3, 110, 111, 120
65, 67, 68, 69, 82, 107, 108, 109, 110, skin, 16, 29, 93
111, 112, 114, 115, 116, 119, 121, 122, skin diseases, 16
123, 124, 125, 126, 127, 128, 129 sludge, 76, 79, 84, 85, 96
rosiglitazone, 121 smooth muscle cells, 125
rotavirus, 14 soapnuts, v, vii, ix, 91, 92, 93, 95, 97, 102,
Royal Society, 84 106
RTS, 6, 8, 33 sodium, 54, 69, 74, 77, 81
142 Index
sodium dodecyl sulfate (SDS), 54, 81 Sun, 6, 8, 9, 10, 19, 20, 28, 30, 31, 32, 33,
soil particles, 74 34, 35, 48, 51, 83, 89, 121, 123, 126, 127
soil remediation, ix, 64, 67, 68, 74, 78, 80, supervision, 54
82, 92, 93, 105 suppression, 16, 116, 118, 122, 124
soil type, 73, 74 surface properties, 86
soil washing, vii, ix, 64, 73, 74, 75, 77, 78, surface tension, ix, 64, 70, 99, 100, 101, 102
79, 80, 81, 82, 84, 85, 86, 88, 89, 91, 92, surfactant(s), 69, 70, 71, 73, 74, 77, 78, 80,
93, 95, 96, 98, 99, 100, 102 81, 82, 83, 84, 85, 86, 88, 93, 94, 95, 99,
Solanum anguivi, 15, 24 100, 101, 102
solubility, 69, 82, 93, 94 survival, 15, 19, 59, 115, 118
solution, vii, ix, 1, 64, 68, 70, 73, 74, 76, 78, suspensions, 86
79, 80, 81, 83, 86, 91, 92, 93, 98, 101, swelling, 16
102 syndrome, 15, 18, 47
solvents, 81, 82, 86, 95 synergistic effect, 81
sorption, 74, 76 synthesis, 27, 42, 123
South Africa, 127
South America, 6
South Korea, 52, 53 T
Southeast Asia, ix, 107
T cell, 16, 33, 119
soybeans, vii, 1, 92
Taiwan, 75, 85
species, vii, 1, 10, 11, 12, 13, 15, 18, 19, 24,
tau, viii, 2, 19
27, 35, 65, 66, 67, 68, 69, 109, 116
techniques, 4, 67, 95
spectrophotometric method, 4
technologies, 4, 82, 84
spectroscopy, 95
technology, 77, 80, 88, 93, 94, 95, 102, 123
spinal cord, 18
temperature, ix, 67, 72, 76, 86, 91, 94, 98,
spleen, 16
102
stability, ix, 58, 64, 69, 80, 94
tension(s), ix, 63, 68, 69, 70, 71, 74, 94, 99,
stabilization, 84
100, 101
stem cells, 48, 53, 56, 59, 60
Terminalia arjuna, 19, 35
steroidal saponins, vii, 1, 2, 3, 12, 25, 28,
territory, 92
66, 109
testosterone, 122
steroids, 108
Th1 immune response, 5, 6
sterols, 12, 13, 128
Th2 response, 6
stimulation, 29, 44, 120
therapeutics, 47
stimulus, 15
thorium, 84
streptococci, 125
threonine, 42
stroke, 20, 28
thrombin, 121
structural changes, 112
tissue, 39, 69
structure, ix, 3, 4, 7, 10, 12, 17, 21, 30, 32,
TLR, 24
34, 42, 43, 48, 50, 63, 64, 67, 69, 92,
TLR9, 29
108, 117, 121, 122
TNF-α, 16, 17, 115
sucrose, 81
tobacco, 117, 129
sugar beet, vii, 1, 65
toxic effect, 66
sulfate, 69
toxicity, 6, 33, 72, 75, 77, 82, 83, 93, 94,
sulfur, x, 108
122, 129
Index 143
toxicology, 127
toxin, 9
V
transcription, 39, 40, 41, 45, 49, 50, 120
vaccine, viii, 2, 6, 8, 9, 21, 22, 23, 24, 25,
transcription factors, 39, 40, 41, 50, 120
27, 28, 29, 30, 31, 32, 33, 34, 35
transformation, 88
vaccine design, 29
transforming growth factor (TGF), 40, 43,
Valencia, 84
47, 49, 51, 52
vancomycin, 124
translocation, 50
vegetables, 92
transplantation, 53, 60, 120
vein, 43
transport, 49
viral infection, 6, 26
trauma, viii, 2, 19
virus infection, 28
treatment, x, 10, 12, 13, 14, 16, 17, 18, 19,
virus replication, 117
25, 38, 43, 44, 45, 67, 73, 74, 76, 80, 84,
viruses, 6, 13, 14
94, 107, 108, 118, 120, 122
viscosity, 80, 94
triglycerides, 39, 44
vitamins, 38
triterpenic saponins, 3
triterpenoid(s), v, vii, x, 1, 2, 3, 9, 10, 11,
12, 13, 16, 21, 22, 23, 24, 25, 27, 28, 30, W
31, 34, 35, 66, 67, 97, 107, 108, 109,
110, 111, 112, 115, 117, 122, 123, 124, washing procedures, 75
125, 126, 127, 128, 129 Washington, 88
tropical forests, 65 waste, 66, 67, 76, 77, 78, 88, 89
TS, 53, 66, 122 wastewater, 99, 100, 101, 106
tuberculosis, 8, 116, 125, 128 water, vii, ix, 2, 3, 42, 43, 59, 69, 70, 72, 75,
tumor, 17, 115, 118 76, 81, 84, 88, 89, 91, 92, 93, 94, 95, 96,
tumor necrosis factor (TNF), 6, 16, 17, 115 98, 101, 102, 115, 119, 122, 123, 126,
tundra, 65 130
type 2 diabetes, 45 wells, vii, ix, 91, 94
tyrosine, 20, 121, 126, 127, 129 wetting, ix, 64, 67, 68, 70, 93
tyrosine hydroxylase, 20 WHO, 66
Wnt signaling, 41, 46, 50
worldwide, 93
U
ulcerative colitis, 126 Y

ultrasound, 4
United Kingdom, 104 Yucca schidingera, 11
United States (USA), 43, 51, 52, 53, 66, 88
upper respiratory tract, 108
uranium, 84 Z
urban, 82
urine, 54, 58 Ziziphys, 11
ursane type triterpenoid, 112 zoospore, 10
Uruguay, 8

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BIOCHEMISTRY RESEARCH TRENDS

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Library of Congress Cataloging-in-Publication Data

Published by Nova Science Publishers, Inc. † New York

Saponins are surfactive compounds that are widely distributed in nature,

are biosynthesized by monocotyledons, such as members of the Liliaceae,

Chapter 3 - Saponins represent glycosides, secondary plant metabolites.

used as a Traditional Chinese Medicine (TCM) for the treatment of headaches,

SAPONINS: OCCURRENCE IN NATURE

Andresa Heemann Betti, PhD, Juliane Deise Fleck, PhD,

aubergine, tomato seed, leek, onion, garlic and asparagus. The

Keywords: triterpenoid saponins, steroidal saponins, activity in vitro, activity

saponins have been attributed to different needs of protection against

Figure 2. Triterpenic aglycones frequently found in plants: (A) β-amyrin (oleanane-

The saponins generally occur in plants as a mixture of diverse compounds

Recently, UPLC coupled with electrospray-ionization quadrupole time-of-

2. IMMUNOLOGICAL ADJUVANT AND

a) to enhance the immunogenicity of highly purified or recombinant

cytokines IL-2, TNF and interferon gamma (INF-), and an enhanced

Figure 3. QS-21 structure.

The saponins from Q. saponaria are glycosylated triterpenes, classified as

a) the choice of lipid composition,

healthy volunteers, evidenced by improvements in PMN cell chemotaxis,

3.1. Haemolytic Activity

3.2. Antifungal Activity

The antifungal/antiyeast activity has been reported since 1993, when

3.3. Antimicrobial Activity

Antimicrobial activity has been reported for triterpenoid saponins since

saponins from Capsicum annuum seeds. They showed weak or no growth

3.4. Antiviral Activity

Antiviral activity of ginsenosides, saponins from ginseng, was evaluated.

3.5. Hypocholesterolemic Activity

Saponins have been studied due to their lowering cholesterol effects, an

cholesterol. Other mechanisms involved in the modulation of the expression of

4. ANTIINFLAMATORY AND ANTIOXIDANT ACTIVITIES

4.1. Anti-Inflammatory Activity

Sepsis is a major clinical challenge in modern medicine, representing one

conditions. The treatment with astragaloside IV significantly decreased the

4.2. Antinociceptive Activity

Ahn et al. (2016) evaluated the antinociceptive effect of ginsenoside Rg3,

4.3. Antioxidant Activity

Saponins have a well-documented antioxidant effect in vitro and in vivo.

Li et al. (2009) indicated that the neuroprotective effect of total saponins

4.4. Neuroprotective Activity

The neurprotective effects of saponins on diseases and injury of the

coronary heart disease, ischemic cerebrovascular disease and hemorrhagic

Augustin, JM; Kuzina, V; Andersen, SB; Bak, S. Molecular activities,

Chen, F; Eckman, EA; Eckman, CB. Reductions in levels of the Alzheimer’s

responses to bovine herpesvirus type 1 in mice. Vaccine., 2006, 24 (49-

Hostettmann, K; Marston, A. Saponins: chemistry and Pharmacology of

Kim, YJ; Wang, P; Navarro-Villalobos, M; Rohde, BD; Derryberry, J; Gin,

Saikosaponin b2 is a naturally occurring terpenoid that efficiently inhibits

Mao, S; Yang, G; Li, W; Zhang, J; Liang, H; Li, J; Zhang, M. Gastroprotective

Oda, K; Matsuda, H; Murakami, T; Katayama, S; Ohgitani, T; Yoshikawa, M.

Skene, CD; Sutton, P. Saponin-adjuvanted particulate vaccines for clinical use.

response to ovalbumin and its haemolytic activities. Vaccine, 2008,

Wang, S; Kennedy, JS; West, K; Montefiori, DC; Coley, S; Lawrence, J;

Xiong, H; Zheng, Y; Yang, G; Wang, H; Mei, Z. Triterpene saponins with

Yong-Seob Jeong1 and Jisun Oh2,

Keywords: ginseng, ginsenoside Rg3, anti-obesity, adipogenesis, lipid

3. REGULATION OF LIPID METABOLISM