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267
Saponins
*Corresponding author: Jacopo TROISI laboratorio.troisi@na.camcom.it
:
J. TROISIa*, R. DI FIOREa, C. PULVENTOb, R. D’ANDRIAb, ANTONIO VEGA-GÁLVEZc, MARGARITA
MIRANDAc, ENRIQUE A. MARTÍNEZd , A. LAVINI b
a Laboratorio Chimico Merceologico, Az. Spec. CCIAA, Corso Meridionale 58, I-80134 Napoli, Italy.
b CNR – Instute for Agricultural and Forest Mediterranean System (ISAFoM)), Ercolano (NA), Italy
c Universidad de La Serena, Facultad de Ingeniería, Av. Raúl Bitrán s/n, Box 599, La Serena, Chile.
d Centro de Estudios Avanzados en Zonas Áridas, CEAZA, Avda. Raúl Bitrán s/n, La Serena, Chile.
The term saponin comes from the Lan word sapo,
meaning “soap”, reecng a readiness to form stable
soap-like foams in aqueous soluons. The biological
role of saponins is not completely understood, but
they are generally considered to be part of a plant’s
defence system against pathogens and herbivores,
parcularly because of their bier avour. Saponins
comprise aglycones and sugar, each represenng
about 50% of the total weight of the molecule. In
quinoa, saponins are a complex mixture of triter-
pene glycosides that derive from seven aglycones:
oleanolic acid, hederagenin, phytolaccagenic acid,
serjanic acid, 3β-hydroxy-23-oxo-olean-12-en-28-
oic acid, 3β-hydroxy-27-oxo-olean-12-en-28-oic
acid and 3β,23α,30β-trihydroxy-olean-12-en-28-oic
acid, while the most common sugars are arabinose,
glucose and galactose. Saponins are tradionally
considered very annutrional because of their
haemolyc acvity, and there is therefore a long-
standing controversy about their funcons in food.
It is believed that saponins can form complexes
with membrane sterols of the erythrocyte, causing
an increase in permeability and a subsequent loss
of haemoglobin. However, recent extensive studies
of the biological acvity of saponins in vitro and in
vivo have idened associaons with several health
benets, including an-inammatory, ancarcino-
genic, anbacterial, anfungal and anviral eects.
Saponins are also of interest as valuable adjuvants
and the rst saponin-based vaccines have been in-
troduced commercially. Tradionally, quinoa seeds
are either abraded mechanically to remove the
bran – which is where the saponins are predomi-
nantly located – or washed with water to remove
bierness prior to use. During washing, valuable
nutrients are lost and the chemical composion and
amino acid proles of quinoa seeds can be altered.
Following treatment, the level of saponin content
in to-be-consumed quinoa seeds remains a major
concern in terms of bierness and possible nega-
ve biological eects. A mathemacal model based
on Fick’s second law has been created to opmize
the leaching process of saponins from quinoa seeds
during washing with water.
Many studies have focused on the eects of agro-
nomic variables (e.g. irrigaon and salinity) on the
saponin proles of quinoa. It has been observed
that saponins decrease in samples that have been
exposed to drought and saline regimes – suggesng
that irrigaon and salinity may regulate the saponin
content in quinoa and aect its nutrional and in-
dustrial values.
Studies are underway to evaluate and compare
the saponin content in seven variees of quinoa
grown in Italy and six variees grown in Chile un-
der rainfed or low irrigaon condions. Seeds from
the more arid or stressing Chilean localies have a
higher saponin content.
268
1.1 Saponin chemistry
Saponins are compounds found in many plants
(Sparg et al., 2004) and they have the disnc-
ve feature of forming foam. The name probably
comes from the plant Saponaria whose roots were
historically used to make soap (Lan sapo = soap)
(Augusn et al., 2011). Chemically, they are glyco-
sides with a polycyclic aglycone (glycoside-free por-
on), which may occur in the form of a steroid or
a triterpenoid choline bound via the C3 carbon by
means of an ethereal bond to a side sugar chain.
The aglycone is commonly referred to as sapogenin,
while the subset of steroidal saponins is commonly
referred to as sarapogenin. Saponins are amphip-
athic because of their fat-soluble aglycone funcon
and their water-soluble saccharide chain. This char-
acterisc is the basis of the ability to form foam.
Saponins are perceived as bier, and this reduces
the organolepc characteriscs and the palatability
of any products rich in them. Only a few (usually
those with a triterpenoic aglycone) have a nice a-
vour, reminiscent of liquorice root.
1.2 Saponin Biosynthesis
Evidence that the overexpression of squalene syn-
thase may induce an up-regulaon of saponins and
phytosterols (Lee et al., 2004) suggests that this
enzyme is involved in the branching of biosynthec
pathways leading to the synthesis of phytosterols
and saponins. This observaon led to the theory
(now consolidated) that saponins derive from the
same anabolic process that leads to the formaon
of phytosterols. All terpenoids derive from con-
densaon of 5-carbon building blocks designated
IPP (3-isopentenyl pyrophosphate) and DMAPP
(dimethylallyl pyrophosphate). In plants, IPP and
DMAPP dri from condensaon of acetyl-CoA in
the mevalonate pathway or from pyruvate and
phosphoglyceraldehyde. Terpenoid biosynthesis in
plants is extensively compartmentalized: steroids,
triterpenes and saponins are mainly synthesized in
the cytosol ulizing IPP from the mevalonate path-
way.
Flores-Sanchez et al. (2002) conducted experiments
in which the acvity of HMG-CoA reductase – a key
enzyme in mevalonate and squalene synthesis –
was inhibited, and this led to a reducon of phy-
tosterols and of ursolic/oleanolic acid biosynthesis,
conrming the hypothesis that the biosynthec
pathway of saponins is linked to that of plant sterols
by means of squalene synthesis.
IPP and DMAPP undergo condensaon to the
10-carbon intermediate GPP (geranyl pyrophos-
phate), and the addion of a second IPP unit leads
to FPP (farnesyl pyrophosphate, C15), the common
precursor of the vast array of sesquiterpenes pro-
duced by plants. Linkage of two FPP units leads to
formaon of squalene (C30). This is then epoxygen-
ated to 2,3-oxidosqualene (C30), considered the
last common precursor of triterpenoid saponins,
phytosterols and steroidal saponins. The steps at
which steroidal saponin and phytosterol biosynthe-
sis diverge have not been elucidated, although Ka-
linoswska et al. (2005) suggest that cholesterol is a
precursor of steroidal saponins.
Summarizes the seven aglycones idened so
far in the dierent parts of quinoa (owers, fruits, seed-
coats and seeds) (Kuljanabhagavad et al., 2008). These
structures have been obtained by means of extensive
characterizaons in NMR (nuclear magnec resonance)
and mass spectrometry. Most of the variability is gener-
ated by the saccharide side chains – indeed, the seven
aglycones give birth to more than 20 saponins (Table 1).
CHAPTER: 3.3 SAPONIS
269
Saponins derived from the 7 aglycones found in quinoa.
und Sugar side chain Aglycone
β-D-Glc(1→3)-α-L-Ara
I
II
III
IV
V
VI
VII
α-L-Ara
III
V
VI
β-D-GlcA
III
IV
VI
β-D-Glc(1→2)-β-D-Glc(1→3)-α-L-Ara
III
IV
V
β-D-Xyl(1→3)-β-D-GlcA IV
β-D-Glc(1→3)-β-D-Gal V
VI
β-D-Glc(1→4)-β-D-Glc(1→4)-β-D-Glc V
The rst commied step in the biosynthesis of triter-
penoid saponins and phytosterols is the cyclizaon
of 2,3-oxidosqualene. During this process, internal
bonds are introduced into the oxidosqualene back-
bone, resulng in the formaon of predominantly
polycyclic molecules containing varying numbers of
5- and 6-membered rings. The high number of pos-
sibilies for establishing dierent internal linkages
during cyclizaon gives rise to a vast array of diverse
structures, and over 100 dierent triterpene skel-
etons have been found in nature. However, from
this vast range, only a limited number of possible
cyclizaon products appear to be ulized in saponin
biosynthesis.
Following the formaon of basal sapogenin back-
bone structures, these common precursors usually
undergo various modicaons prior to glycosyla-
on. The most common sapogenin modicaons
are small funconal groups, such as hydroxyl-, keto,
aldehyde - and carboxyl-moiees at various posi-
ons of the backbone.
Glycosylaon paerns of saponins are oen con-
sidered crucial for their biological acvies. Typical
triterpenoid saponin glycosylaon paerns consist
of oligomeric sugar chains of 2–5 monosaccharide
units, most oen linked at posions C3 and/or C28.
Less oen, 1–2 monosaccharide units have been
reported to occur at posions C4, C16, C20, C21,
C22 and/or C23. Glucose, galactose, glucoronic
acid, rhamnose, xylose and arabinose are the most
abundant hexoses and pentoses in the saccharide
chains. Saponin glycosylaon presumably involves
sequenal acvity of dierent enzymes belonging
to the mulgene family of uridin diphosphate gly-
cosyltransferases (UGTs).
CHAPTER: 3.3 SAPONIS
270 1.3 Biological role
Saponins have dierent biochemical acvies.
Francis et al. (2002) reported, among others, strong
haemolyc, anmicrobial, fungicidal, allelopathic,
inseccidal and molluscicidal acvity, while Vega-
Gálvez et al. (2010) reported their eects as a vac-
cine coadjuvant. Therefore, although the true bio-
logical signicance of saponins in quinoa sll needs
to be fully determined, the current line of thought
is that they are part of the plant’s apparatus to de-
fend o predators.
1.3.1 Haemolyc acvity
One of the systems used to probe the presence of
saponins in a plant extract or in a drug is based on
incubaon of the extract with blood red cells and
vericaon of the degree of haemolysis of the sam-
ple. The ability of saponins to break the membrane
of the erythrocytes is linked to their ability to bind
membrane sterols (Khalil et al., 1994). When the
membrane bursts, there is an increase in perme-
ability and a loss of haemoglobin. Baumann et al.
(2000) have invesgated the eect of saponins on
the membrane structure through haemolysis of hu-
man erythrocytes. The ndings show that saponin-
lysed erythrocytes do not reseal, indicang that
saponin-induced damage to the lipid bilayer is irre-
versible. The level of haemolyc acvity has been
aributed to the type of aglycone and to the pres-
ence of the sugar side chains (Wang et al., 2007).
1.3.2 An-inammatory acvity
In the carrageenan-induced oedema assay, many
saponins isolated from plant sources produce an
inhibion of inammaon. Kim et al. (1999) sug-
gested that the an-inammatory acvity of these
saponins is related to ancomplementary ac-
on through the classical inammaon pathway.
Oleanolic acid and ginsenoside Ro show the highest
ancomplementary acvity.
1.3.3 Anfungal/anyeast acvity
Triterpenoid saponins from the seeds of Chenopo-
dium quinoa Willd. (Chenopodiaceae) have been
reported to have anfungal acvity (Woldemichael
and Wink, 2001). A study by Bader et al. (2000)
revealed that the anfungal acvity of saponins
against dierent Candida albicans strains can be
inuenced by variaon of the etherglycosidically
bonded carbohydrate units and the acylglycosidi-
cally bonded oligosaccharide at C-28 of the agly-
cone. However, only crude saponin mixture inhibits
the growth of Candida albicans. Pure compounds
show lile or no acvity, which suggests a possible
synergisc eect between these saponins.
1.3.4 Anbacterial/anmicrobial acvity
Saponins have also been reported to have anmi-
crobial acvity (Killeen et al., 1998). Alcohol soluble
saponins have anmicrobial acvity towards both
prokaryoc and eukaryoc organisms, but only at
low cell densies, and they do not inhibit microbial
growth of dense populaons.
1.3.5 Cytotoxicity and antumour acvity
Numerous reports highlight the highly cytotoxic
properes of many saponins (Musende et al., 2009;
Man et al., 2010). In parcular, oleananes show
an antumour eect in various pathways, includ-
ing ancancer, anmetastasis, immunosmulaon
and chemoprevenon. The detailed mechanisms
are complex but involve dephosphorylate Stat3 in a
variety of human tumour cell lines and lead to a de-
crease in the transcriponal acvity of Stat3, which
regulates proteins such as c-myc, cyclin D1, Bcl2,
survivin and VEGF. Moreover, several immunosm-
ulang acvies, such as induced growth of human
T lymphocytes, promong apoptosis and triggering
autophagic cell death have been reported. They
decrease respiratory acvity and induced ATP ef-
ux aer inhibion of the voltage-dependent anion
channel in the outer mitochondrial membrane.
Saponins are generally bier, so before consump-
on they must to be eliminated from quinoa. Tra-
dionally, quinoa seeds are either mechanically
abraded to remove the bran, where the saponins
are predominantly located, or washed with water to
remove bierness prior to use. Wright et al. (2002)
report that during this washing process, valuable
nutrients are also lost and the chemical composi-
on and amino acid proles in quinoa seeds may be
altered. The nal level of saponin content in to-be-
consumed quinoa seeds remains a major concern
in terms of its bierness and possible negave bio-
logical eects.
CHAPTER: 3.3 SAPONIS
271
2.1 Kinec
The removal of saponins from quinoa seeds during
washing can be described according to the rules
governing solid–liquid extracon and by applying
mathemacal models generally used to evaluate
process kinecs.
The total saponin concentraon inside quinoa
seeds rapidly tends towards an asymptoc value
following an inial leaching. Fuentes et al. (2013)
show that this asymptomac value decreases as the
washing temperature increases.
Saponin rao (SR) – dened according to equaon
1 – is the most commonly used parameter for mod-
elling the saponin leaching kinecs of quinoa seeds.
SR represents a dimensionless concentraon used
to study the leaching kinecs, supposing a mecha-
nism of diusion inside the solid and negligible ex-
ternal mass transfer under condions of intensive
srring.
SR= Xst-Xse Eq.1
Xs0-Xse
where Xst is the saponin content in real me
(g/100gdm), and Xs0 and Xse are the inial and residu-
al saponin contents.
Table 2 represents the most important model
adopted for modelling SR in saponin removal.
2.2 Uses of Saponins
Saponins are used in industry as addives in foods
and cosmecs. They can also be used in other in-
dustrial applicaons (Yang et al., 2010; Chen et al.,
2010; Price et al., 1987; Hostemann and Marston,
1995) as, for example, preservaves, avour modi-
ers, detergents (due to their chemical properes
and abilies as foaming agents) and agents for cho-
lesterol removal from dairy products.
Notably, saponins can also acvate the mamma-
lian immune system, arousing signicant interest
in their potenal as vaccine adjuvants (Sun et al.,
2009). Their unique capacity to smulate both the
Th1 immune response and the producon of cyto-
toxic T-lymphocytes (CTLs) against exogenous an-
gens makes them ideal for use in subunit vaccines
and vaccines directed against intracellular patho-
gens, as well as in therapeuc cancer vaccines.
3.1 Analycal methods
Several analycal methods have been developed for
the determinaon of saponins from various matri-
ces, including quinoa seeds. The simplest methods
are used to detect typical saponin features, such as
their ability to form foam or their haemolyc abil-
ity. The most commonly used methods, however,
are chromatographic. Both liquid chromatography
(with detecon by mass spectrometry, DAD and
: Mathemacal models selected to describe saponin leaching kinecs
Model Equaon Reference
Vega-Gàlvez et al.
(2011)
Corzo et al. (2008)
Logarithmic Akpinar (2006)
Sacilik & Elicin (2006)
Two terms Lahsasni et al. (2004)
Tog˘ rul& Pehlivan
(2003)
CHAPTER: 3.3 SAPONIS
272 ELSD), and gas chromatography (with detecon by
mass spectrometry and FID) have been employed.
Gas chromatography has been widely used, al-
though providing for a longer extracon protocol
and a delicate silanizaon reacon. The rst studies
to include determinaon by gas chromatography
were those by Ridout et al. (1991) and Price et al.
(1986). In gas chromatography, saponins are gener-
ally extracted aer acid hydrolysis of the degrased
sample with a polar solvent; the extract aer silani-
zaon is analysed with non-polar or slightly polar
columns and eluted at high temperatures. The anal-
ysis in HPLC, on the other hand, entails a simpler
preparaon consisng of extracon with alcohols
and puricaon with a C18 SPE. Separaon is usual-
ly achieved with C18 staonary phases and eluons
in water-acetonitrile gradient, both for photometric
detecon (DAD, ELSD) and in mass spectrometry.
3.2 Saponin evaluaon in Chilean quinoa ecotypes
3.2.1. Ecotypes present in Chilean quinoa agro-
ecological regions
Five quinoa ecotypes are described for the Andean
region. They come from the Inter-Andean valleys
of Colombia, Ecuador and Peru, the Alplano of
Peru and Bolivia, Yunga in the Bolivian subtropical
forest, Salare (salt ats) in Bolivia, Chile and Argen-
na, and the Coastal (lowlands) or sea level areas of
Chile. Their origins and possible expansion routes
have been reviewed by Fuentes et al. (2012). In
Chile, just two of the ve ecotypes have been found
(Salare and Coastal). However, within these two
ecotypes many landraces or local farmers’ variees
exist in the country. In the Alplano (highlands) at
4 000 m asl (19°S), farmers hold at least 12 of these
landraces (Alfonso, 2008; Alfonso and Bazile, 2009),
known by the local Aymara people as, for example,
‘Pandela’ (red seeds), ‘Jankú’ (white seeds), ‘Churi’
(yellow seeds), ‘Chullpe’ (brown seeds), ‘Khánchi’
(dark pink seeds) and ‘Chále’ (mixed colours). In
central (34°S) and southern (39°S) Chile, the lan-
draces appear less abundant because there is less
diversity of seed colour, as most are whish, yellow-
ish, beige and grey, the laer being more abundant
at southern latudes (39°S), as is also observed in
seed bank collecons used for tesng comparave
yields (Marnez et al., 2007).
Of these three regions, the climac condions are
more stressful in the high Andes of northern Chile
where annual rainfall is 100–200 mm (Lanino,
2006), while in central and southern Chile, it is over
400 mm (Miranda et al. 2013).
3.2.2 Saponin content
The total saponin content evaluated in whole seeds
of Chilean landraces and in one hybrid variety (‘Re-
galona’) is over 1%. They are, therefore, all bier
(i.e. saponins > 0.11%) but with signicant varia-
on among them. Unexpectedly, high Andes Sal-
are landraces do not always contain higher values
of saponins (2%). Those from central Chile have the
highest values, reaching as much as 4% (Miranda et
al., 2012). When seeds are sown in a dierent lo-
cality, parcularly culvated under the drier condi-
ons of arid Chile (at 30°S with no rainfall between
October and May), harvested seeds increased their
saponin content, at least for the ‘Regalona’ hybrid,
from 2.2% to 3.2%. This phenomenon, however, is
not observed for another landrace from Villarrica in
southern Chile. The laer maintains a saponin con-
tent of 2.11–2.38% when culvated in arid north-
ern Chile (Miranda et al., 2013). The higher saponin
content in landraces from central Chile might be due
to the parcular stressing condions of high salin-
ity in some coastal soils. These soils are somemes
naturally irrigated in the winter with brackish waters
from the neighbouring rivers inuenced by the high
des of the Pacic Ocean (Orsini et al., 2011).
3.2.3 Conclusions
1. Saponin content has to date been studied in
seeds from Chilean landraces of quinoa belong-
ing to the Salare and Coastal Andean ecotypes.
Their saponin content is high (> 2%), compared
with some sweet quinoas of the Alplano (<
0.11%).
2. Unexpectedly, saponin content is higher in
coastal landraces from central Chile
3. The saponin content of some quinoa seeds
changes when grown under dierent condi-
ons, normally increasing in a more stressing
climate (drought).
3.3 Italian research acvity
From 2006, dierent eld trials have been per-
formed at ISAFoM-CNR to test quinoa. The strategic
objecves of these studies have been: to evaluate
CHAPTER: 3.3 SAPONIS
273
the quantave and qualitave responses of qui-
noa accessions under combined abioc stresses
(salt and drought stress) and their adaptability in
the Mediterranean environment of southern Italy
(see Chapter 6.3); to improve food producon by
introducing quinoa as a possible alternave crop for
this area (potenally high value food cash crops);
and to verify the opportunies for use of quinoa
seeds, ours and derivaves in product lines for
children and for people with coeliac disease, with
potenally interesng growth prospects in special-
ized sectors.
At the experimental staon of the Naonal Re-
search Council (CNR), Instute for Agricultural and
Forest Mediterranean Systems (ISAFoM) in Vitulazio
(CE) (14°50’E, 40°07’N, 25 m asl), a 2-year (2006–
07) eld trial was carried out to compare two qui-
noa genotypes: ‘Ticaca’ (‘KVLQ52’) and ‘Regalona
Baer’ (‘RB’) under rainfed condions (Pulvento et
al., 2010). Comparison was also made between two
sowing dates (April and May) for ‘KVLQ52’ (‘KV’april
and ‘KV’may). In this period, quinoa was studied with-
in the project “CO.Al.Ta. II” (Alternave Crops to To-
bacco), set up by the European Community (CE), to
explore the possibilies of diversicaon of Italy’s
tradional tobacco-growing areas and to evaluate
seed quality, and in parcular saponin content, in
collaboraon with the Department of Food Tech-
nology (DISTAAM) of the University of Molise.
Results show that April is the best sowing me for
quinoa in the Mediterranean region (Table 2). Of
the two genotypes, ‘RB’ records beer growth and
producvity, apparently being more tolerant to abi-
oc stress (high temperatures associated with wa-
ter stress).
The study includes quantave/qualitave assess-
ment of saponins. Gas chromatography analysis
shows that the two variees of quinoa are in an
intermediate posion between “sweet” and “bit-
ter” genotypes. In parcular, the total saponin con-
tent of 238.9 and 213.8 mg/100 gdm for genotype
‘KV’april (sown in April) and ‘KV’may (sown in May),
respecvely, was obtained. For genotype ‘RB’, the
saponin content is 328 mg/100 gdm. From a quali-
tave point of view, conrmed by bibliographic
data (Ridout et al., 1991), oleanolic acid is the main
saponin component (76–85%), followed by heder-
agenin (10–18%) and phytolaccagenin (4–5%).
Since saponins are mainly located in the outer lay-
ers of the seed, these components were removed
through the process of pearling. The process was
performed using a laboratory perlator model (TM-
05-Takayama, tesng Mill) with an abrasive roller
(40P). A 50% reducon in total saponins % com-
pared with the inial value for the product with a
pearling degree of 20% was observed by gas chro-
matographic analysis. However, the nal product
sll had a saponin content which could be detected
at sensory level. Applicaon of pearling at 30% re-
duced the saponin content by about 80%. In fact,
saponin values dropped from 238.9 mg/100 gdm to
33.47 mg/100 gdm in the pearled product (Table 3).
Ash, protein and lipid content in ‘Ticaca’ is higher
aer abrasion of the pericarp. In parcular, the lin-
oleic omega fay acid is very high in ‘Ticaca’ seed
and our.
Seed abrasion tends also to increase oleic, linoleic
and palmic fay acid in ‘Ticaca’.
From 2008 to 2013, ISAFoM-CNR parcipated as a
partner in the UE project “Sustainable water use se-
curing food producon in dry areas of the Mediter-
ranean region” (SWUP-MED).
Quinoa genotype ‘Q52’ (‘Ticaca’) was grown in
an open eld trial in 2009 and 2010 to invesgate
: Saponin content (mg/100 gdm) in the two accessions
Accession Total saponin
Oleanolic ac.
Hederagenin Phytolaccagenin
mg 100 g
-1
of DW % of total saponin
KV
ap ril
238.9 ± 10.87 78.2 16.7 5.1
KV
may
213.8 ± 7.52 76.3 18.9 4.8
RB 329.0 ± 6.78 85.3 10 4.7
CHAPTER: 3.3 SAPONIS
274 the eects of salt and water stress on quantave
and qualitave aspects of the yield. Treatments ir-
rigated with well water (‘Q100’, ‘Q50’ and ‘Q25’)
and corresponding treatments irrigated with saline
water (‘Q100S’, ‘Q50S’ and ‘Q25S’) with an electri-
cal conducvity (ECw) of 22 dS/m were compared.
Saline and water stress in both years do not cause
signicant yield reducon, and quinoa may be de-
ned as tolerant to salinity and drought (Pulvento
et al., 2012).
Chemical composion of quinoa seeds conrms a
higher protein and bre content compared with
common cereals, while the highest level of saline
water determines higher mean seed weight and, as a
consequence, higher bre and total saponin content
in quinoa seeds. It has been observed that irrigaon
with 25% full water restuon, with and without the
addion of salt, is associated with an increase in free
phenolic compounds of 23.16% and 26.27%, respec-
vely. In contrast, bound phenolic compounds are
not aected by environmental stresses.
The eects of the dierent agronomic variables,
such as irrigaon and salinity, on the saponin pro-
les of quinoa were analysed.
Saponins were evaluated in terms of sapogenins
(Gomez-Caravaca et al., 2012; Lavini et al., 2011)
(Figure 2).
A gas chromatographic procedure was applied for
the evaluaon of saponin aglycones (sapogenins)
derived from the acid hydrolysis of samples (Ridout
et al., 1991; Woldemichael and Wink, 2001). Three
major quinoa saponin aglycones were idened:
oleanolic acid (36–50% total), hederagenin (27–
28%) and phytolaccagenic acid (21–36%) (Figure 3).
Schemac diagram for the extracon of
saponins
Soxhlet Extracon
GC analisis
Sample
Defaed sample
Sapogenin
Quantave and qualitave
evaluaon of sapogenis
0
50
51
52
53
54
55
56
57
58
59
60
61
62
50
50
51
52
53
54
55
56
57
58
59
60
61
62
2
4
6
8
10
12
14
16
18
20
22
24
26
28
30
1
2
3
1
2
3
GC chromatogram of Ticaca saponin (1 Oleanolic acid, 2 Hederagenin, 3 Phytolaccagenic acid)
CHAPTER: 3.3 SAPONIS
275
When considering the total amount of saponins
(Table 5) it was observed that ‘Ticaca’ is a bier
variety. In fact, quinoa seeds with a saponin con-
centraon > 0.11% are usually considered to be bit-
ter genotypes (Vega-Gálvez et al., 2010).
The highest saponin values were observed in sam-
ples obtained without decit irrigaon treatments
(1 633.3 mg/100 g for ‘Q100S’ and 1 140.1 mg/100
gdm for ‘Q100’, respecvely). The samples treated
with a water decit (‘Q25’ and ‘Q50’) showed a de-
crease in saponin content compared with ‘Q100’.
The ‘Q50’ samples, compared with ‘Q100’, showed
a decrease in saponins of 32%; while the sam-
ples grown with a higher irrigaon decit (‘Q25’)
showed a 45% decrease in saponins. These results
are in agreement with the study of Soliz-Guerrero et
al. (2002), who reported that saponin content is af-
fected by a soil-water decit, to the extent that high
water decits promote low saponin contents. Sam-
ples treated with saline water also show signicant
dierences at dierent irrigaon levels (‘Q100S’,
‘Q50S’ and ‘Q25S’); the decrease in saponin content
in the ‘Q50S’ and ‘Q25S’ samples is very high com-
pared with ‘Q100S’ (40% and 42% for ‘Q25S’ and
‘Q50S’, respecvely).
From 2011 to 2013, eld trials were performed in
Vitulazio on quinoa, and others are ongoing at ISA-
FoM within the “CISIA” project, funded by the Na-
onal Research Council, and the “Quinoa Felix” pro-
ject – Introducon of quinoa (Chenopodium quinoa
Willd.) – in the Campania region for high nutrional
and funconal value food producon, in collabora-
on with the University of Molise and CNR-Instute
of Food Science (ISA) of Avellino. The aim of these
acvies is to evaluate yield and seed quality of
Chenopodium quinoa variees grown under rainfed
condions in southern Italy, and to assess milling
performance and protein, ash, lipid and saponin
content of the seed.
All analyses are performed on whole seeds and on
“pearling” grain, aer removal of the pericarp, to
dene the potenal nutrional characteriscs of
each quinoa variety. Since there is no genec re-
source of quinoa as a domescated variety in Italy,
the studies are conducted using seeds received from
foreign instuons and of dierent origins. Tesng
is being done on the Danish quinoa culvars ‘Puno’
and ‘Ticaca’ selected from material originang in
southern Chile and provided by the University of
Copenhagen; four Bolivian culvars ‘Kurmi’, ‘Janca
grano’ ‘Blanquita’ and ‘Real’; the Peruvian ‘Amarilla
de Marangani’; and ‘Jujuy rosada’ originang in Ar-
genna. The Danish culvars ‘Ticaca’ and ‘Puno’
give the higher yield, while ‘Janca grano’, ‘Real’ and
‘Kurmi’ give the lowest yields; ‘Blanquita’ does not
produce under Mediterranean condions.
All seven aglycones have been assayed. The variety
‘Jujuy Rosada’ is richest in saponins (4.99%), while
‘Real’ is the poorest (0.1%). Although the concen-
traon proles of the seven aglycones vary greatly
among the variees – in parcular, in ‘Jujuy rosada’,
72.5% of saponins contain 3β-hydroxy-23-oxo-olean-
12-en-28-oic acid as aglycone, while in ‘Real’, oleanol-
ic acid is the most represented aglycone (despite only
24.80%) – there is a more homogeneous distribuon
of all seven aglycones. However, 3β,23,30-trihydroxy
olean-12-en-28-oic acid is the least represented agly-
cone in all the variees studied.
Saponins present both an obstacle and an opportu-
nity. The deployment as food of many pseudocere-
als, especially quinoa, is hindered by the presence
of these annutrional elements, both because of
reduced palatability due to their bier taste, and
because of the serious eects they can have on hu-
man health. On the other hand, these molecules
are proving to be extremely interesng in several
elds: from pharmaceucal (as the basis for the de-
velopment of new cancer drugs, new anfungals or
adjuvants in vaccines), to chemical, but especially in
the eld of agronomy, where they are proving to be
excellent and versale inseccides. Saponin insec-
cidal acvity is based on three dierent mecha-
nisms (Chaieb, 2010): interference with feeding, en-
tomotoxicity (various forms of chronic toxicity, such
as female ferlity reducon and decreased rate of
blossoming eggs, are observed in many insect spe-
cies) and growth regulaon (research shows that
saponins are able to regulate the growth of many
insect species). The eects of saponins are generally
associated with disturbance of the developmental
stages and moulng failure. Nevertheless, there is
sll massive scope for understanding and improving
this use of saponins, regarding in parcular: stabil-
ity (because the bulk of inseccide acvity is due to
the sugar side chains and these are very suscepble
to pH values and enzymac acvity), applicaon,
CHAPTER: 3.3 SAPONIS
276 acon of residual saponins and their annutrional
properes, and, nally, their dicult synthesis. The
laer could be solved by means of extracon pro-
tocols from variees that produce large amounts of
saponins or are grown under condions that gener-
ate larger quanes (good water supply and high
salinity of the soil), while knowledge of the pedo-
climac eects on saponin content may allow the
development of variees requiring sustainable ag-
ronomic treatments to eliminate these dangerous
annutrional agents.
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