Pharmaceutical Biology

ISSN: 1388-0209 (Print) 1744-5116 (Online) Journal homepage: http://www.tandfonline.com/loi/iphb20

Phytochemical and pharmacological potential of

Medicago sativa: A review

Kundan Singh Bora & Anupam Sharma

To cite this article: Kundan Singh Bora & Anupam Sharma (2011) Phytochemical and
pharmacological potential of Medicago sativa: A review, Pharmaceutical Biology, 49:2, 211-220,
DOI: 10.3109/13880209.2010.504732

To link to this article: http://dx.doi.org/10.3109/13880209.2010.504732

Published online: 25 Oct 2010.

Submit your article to this journal

Article views: 649

View related articles

Citing articles: 4 View citing articles

Full Terms & Conditions of access and use can be found at

http://www.tandfonline.com/action/journalInformation?journalCode=iphb20

Download by: [201.141.162.51] Date: 17 May 2017, At: 07:26

Pharmaceutical Biology, 2011; 49(2): 211220
2011 Informa Healthcare USA, Inc.
ISSN 1388-0209 print/ISSN 1744-5116 online
DOI: 10.3109/13880209.2010.504732

REVIEW ARTICLE

Phytochemical and pharmacological potential of

Medicago sativa: A review
Kundan Singh Bora1 and Anupam Sharma2
1
L.R. Institute of Pharmacy, Solan, Himachal Pradesh, India, and 2University Institute of Pharmaceutical Sciences,
Panjab University, Chandigarh, India

Abstract
Context: Many herbal remedies have so far been employed for the treatment and management of various ailments
since the beginning of human civilization. Medicago is an extensive genus of the family Leguminosae, comprising
about 83 different species. Medicago sativa (Linn.) has long been used as traditional herbal medicine in China, Iraq,
Turkey, India and America for the treatment of a variety of ailments.
Objectives: The aim of this review was to collect all available scientific literature published and combine it into this
review. The present review comprises the ethnopharmacological, phytochemical and therapeutic potential of
M.sativa.
Methods: The present review includes 117 references compiled from major databases as Chemical Abstracts, Science
Direct, SciFinder, PubMed, Dr. Dukes Phytochemical and Ethnobotany, CIMER, and InteliHealth.
Results: An exhaustive survey of literature revealed that saponins, flavonoids, phytoestrogens, coumarins, alkaloids,
amino acids, phytosterols, vitamins, digestive enzymes and terpenes constitute major classes of phytoconstituents
of this plant. Pharmacological reports revealed that it is used as neuroprotective, hypocholesterolemic, antioxidant,
antiulcer, antimicrobial, hypolipidemic, estrogenic, and in the treatment of atherosclerosis, heart disease, stroke,
cancer, diabetes and menopausal symptoms in women.
Conclusion: M. sativa seems to hold great potential for in-depth investigation for various biological activities,
especially their effects on central nervous and cardiovascular system. Through this review, the authors hope to attract
the attention of natural product researchers throughout the world to focus on the unexplored potential of M. sativa,
and it may be useful in developing new formulations with more therapeutic value.
Keywords: Antidiabetic, antiatherosclerotic, hypocholesterolemic, Medicago sativa, neuroprotective

Introduction used in India and Malaysia as an antidote for insect and

The use of plants for medicines or tonic properties goes
back to prehistoric times and has attracted the interest
of scientists for centuries. The Vedas give the earliest
written record about the science of healing. The refer-
ence to medicinal plant is also found in the Ebers papy-
rus (16th century B.C.) which lists in detail over 7000
herbal remedies, for example poppy, castor oil, caraway
etc. The Indians were the first to use chaulmoogra fruits
for its antileprotic activity. The Brazilians employed
Ipecacuanha for the treatment of dysentery and diar-
rhea. The root of Rauwolfia serpentine (Benth.), an indig-
enous plant, has since time immemorial been widely
snake bites and mental derangement. The utility of plants
as therapeutic agents in traditional medicine system is
still prevalent today. For example, the Middle Eastern
civilization developed the Greco-Arabic system of medi-
cine (Unani system of medicine), which is practiced in
the Indian sub-continent. The Ayurvedic and Sidha sys-
tem of medicines were contributed by Indians. All these
systems procure more than 80% of their medicaments
from plants (Bhavna etal., 2007).
Medicago sativa Linn. (Leguminosae), commonly
known as the father of all foods (al-fal-fa), is a peren-
nial herbaceous leguminous plant species that originated

Address for Correspondence: Kundan Singh Bora, L.R. Institute of Pharmacy, Solan, Himachal Pradesh 173212, India. Tel.: +91-1792 252854.
Fax: +91-1792 252851. E-mail: kundanresearch1381@gmail.com
(Received 11 June 2010; revised 24 June 2010; accepted 24 June 2010)

211
212 Kundan Singh Bora, and Anupam Sharma
in Asia (Ehsanpour & Razavizadeh, 2005; Duke, 1985;
BHMA, 1996). This is the most ancient plant, cultivated
throughout the world as a fodder plant. In America, M.
sativa has been extensively cultivated since the arrival of
Europeans. M. sativa has been grown for a variety of pur-
poses such as soil improvement, animal feed and medici-
nal uses (Steppler, 1987).
M. sativa has a long tradition of use as Ayurvedic and
homoeopathic medicine in central nervous and digestive
system disorders, and for the treatment of various other
ailments. However, only limited research has been con-
ducted on this plant species. The present review empha-
sizes the traditional uses and phytopharmacological
potential of M. sativa. Additionally, sporadic pharma-
cological work has so far been carried out to prove its
traditional claims. Through this review, the authors hope
to attract the attention of natural product researchers
throughout the world to focus on the unexplored poten-
tial of M. sativa. No review so far has been compiled on
this species, and therefore it was considered important
to take stock of what has been done in the past, in order
to provide a solid foundation and direction to any future
research that may be conducted on this plant.
Scientific classifications (Heywood & Ball, 1968)
Kingdom: Plantae; Division: Magnoliophyta; Class:
Magnoliopsida; Order: Fabales; Family: Fabaceae;
Subfamily: Faboideae; Tribe: Trifolieae; Genus: Medicago;
Species: sativa; Binomial name: M. sativa Linn.
Description: general morphology
M. sativa is a cool season perennial legume living from
three to twelve years, depending on variety and climate.
The general morphology of M. sativa plant was consid-
ered by Teuber and Brick (1988), and Barnes and Sheaffer
(1995). The mature M. sativa plant is characterized by a
strong taproot. This taproot may eventually surpass 6 m
or more in length with several to many lateral roots con-
nected at the crown when M. sativa is grown in deep, well
drained, moist soils. The crown, a complex structure near
the soil surface, has perennial meristem activity, produc-
ing buds that develop into stems. Tri- or multi-foliolate
leaves form alternately on the stem, and secondary and
tertiary stems can develop from leaf axils. A plant in a
typical forage production field has between 5 and 15
stems and can reach nearly 1 m in height. Flowers vary in
color yet purple, variegated, yellow, cream and white are
the most common. After pollination, these flowers most
commonly produce spiral-shaped seed pods.
Worldwide production
M. sativa is the most cultivated legume in the world.
Worldwide production was around 436 million tons
(436000000 metric tons) in 2006. The USA is the largest
M. sativa producer in the world, but considerable area is
 Pharmaceutical Biology
found in Argentina (primarily grazed), Australia, South
Africa, and the Middle East (FAO, 2006). Within the USA,
the leading M. sativa growing states are California, South
Dakota, and Wisconsin. The upper Midwestern states
account for about 50% of US production, the Northeastern
states 10%, the Western states 40%, and the Southeastern
states almost none. M. sativa has a wide range of adap-
tation and can be grown from very cold northern plains
to high mountain valleys, from rich temperate agricul-
tural regions to Mediterranean climates and searing hot
deserts.
Ethnopharmacological uses
Traditionally, M. sativa is used to improve the memory, to
cure kidney pain, cough, sore muscles, as a rejuvenator,
antidiabetic, antioxidant, anti-inflammatory, antifungal,
anti-asthmatic, antimicrobial, diuretic, galactagogue
and in central nervous system (CNS) disorders (Finkler,
1985; BHMA, 1996; Inamul, 2004; DerMarderosian etal.,
2005). M. sativa has a long tradition of use as Ayurvedic
and homoeopathic medicine in CNS disorders (Inamul,
2004). M. sativa has been used by the Chinese since the
sixth century to treat kidney stones, fever, gravel, dysuria
and to relieve fluid retention and swelling. Ancient Indian
Ayurvedic physicians used M. sativa to treat ulcers,
arthritis pain and fluid retention. In Mexico M.sativa is
believed to improve the memory, to cure sore muscles
and inflammation. Early Americans used M. sativa to
treat arthritis, boils, cancer, scurvy, and urinary and
bowel problems. In Iraq M. sativa is used in arthritis. In
Turkey it is used as cardiotonic and to treat scurvy and
arthritis (Michael etal., 2007; Finkler, 1985; Inamul, 2004;
DerMarderosian etal., 2005).
Moreover, it is considered beneficial in bladder dis-
orders, blood clotting disorders, boils, cough, diuresis,
gastrointestinal tract disorders, breast cancer, cervical
cancer, kidney disorders, prostate disorders, appetite
stimulation, inflammation, increasing breast milk,
asthma, indigestion, insect bites, jaundice, menopausal
symptoms, allergies, increasing excretion of neutral ste-
roids and bile acids in fecal matter, nutritional support,
stomach ulcers, skin damage from radiation, galact-
agogue, increasing peristaltic action of the stomach and
bowels, thrombocytopenic purpura, uterine stimulant,
rheumatoid arthritis, scurvy, vitamin supplementation
(vitamins A, C, E, K) and wound healing (Gray & Flatt,
1997a, 1997b; BHMA, 1996; DerMarderosian etal., 2005;
Swanston etal., 1990).
Phytochemical reports
M. sativa has been reported to contain a variety of
phytochemicals. It has following different classes of
phytoconstituents.
Alkaloids: asparagines, trigoneline, stachydrine,
l-homostachydrine (Duke, 1985; Mills, 1994; Tamsyn
etal., 2009). Amino acids: medicanine, lysine, arginine,

histidine, tyrosine, phenylalanine, methionine, aspartic
acid, glutamic acid, asparagine, serine, alanine, threo-
nine (Worthington & Breskin, 1983; Fushiya etal., 1984;
Lihu & Pedak, 1979; Mego & Erdelsky, 1977). Carotene
(Gupta et al., 1981). Coumarins: myrsellinol, scopole-
tin, esculetin, 4-coumaric acid (Duke, 1985; El-Khrisy
et al., 1994; Orr et al., 1993). Digestive enzymes (Duke,
1985). Enzymes: isoflavone reductase, vestitone
reductase, iminopeptidase, and two aminopeptidases
(Xiaoqiang etal., 2006; Shao etal., 2007; Biagioni etal.,
1990). Flavonoids: quercetin, myricetin, luteolin, api-
genin, chrysoeriol, tricin, 4'-O-[2'-O-E-feruloyl-O--
glucuronopyranosyl(12)-O--lucuronopyranoside]
apigenin,7-O--glucuronopyranosyl-4'-O-[2'-O-E-feruloyl-
O--glucuronopyranosyl(12)-O--glucuronopyra-
noside] apigenin, 7-O--glucuronopyranosyl-4'-O-[2'-
O-p-E-coumaroyl-O--glucuronopyranosyl(12)-O--
glucuronopyranoside] apigenin, 7-O-{2-O-E-feruloyl-
[-glucuronopyranosyl(13)]-O--lucuronopyranosyl
(12)-O--glucurono-pyranoside} apigenin, O-glycosyl
transferases, medicarpin, coumestrol, sativan, vestitol,
formononetin (Bickhoff et al., 1964; Stochmal et al.,
2001; Kowalska et al., 2007; Stochmal & Oleszek, 2007;
Parry & Edwards, 1994; Dutu etal., 2002; Dixon & Steele,
1999; Sumner etal., 1996; Piretti etal., 1982). Minerals:
Ca, K, P, Mg, Fe, Zn, Cu, Al, B, Cr, Co, Mn, Mo, Se, Si, Na,
Sn (Worthington & Breskin, 1983; Potgieter, 2005).
Non-protein amino acids: l-canaverin (Albourine et al.,
1991; Tamsyn et al., 2009). Organic acids: citrate, malate,
malonate, succinate, fumarate, lactate, benzoate (Francoise
et al., 1991; Tamsyn et al., 2009). Phenolic compounds:
p-hydroxybenzoic acid, vanillic acid, p-coumaric acid,
ferulic acids, salicylic acid, sinapic acids, caffeic acid,
hesperetin, naringenin, chlorogenic acid, tannic acid,
heterosides (Newby et al., 1980; Volynets et al., 1979;
Dutu etal., 2002). Phytoestrogens: coumestrol, genistein,
formometin, diadzein, biocanine A (Adler, 1962; Bickhoff
et al., 1960). Phytosterols: -sitosterol, stigmasterol
(Timbekova et al., 1996). Polyamines: norspermidine,
norspermine (Rodriguez-Garay et al., 1989). Protein:
ferritin, protein phosphatase 2A holoenzyme, -amylase
(Doehlert et al., 1982; Toth et al., 2000; Barcelo et al.,
1997). Saponins: soyasapogenols, hederagenin, medi-
cagenic acid (Oleszek & Jurzysta, 1986; Massiot et al.,
1988; Oleszek, 1988; Wieslaw et al., 1990), 3-Glc-Glc-
Rha-28Glc-medicagenic acid, 3-Glc-28-Glc-medicagenic
acid, 3-Glc-malonyl-28-Glc-medicagenic acid, Ara-
Glc-Ara-hederagenin, 3-Glc-Ara-28-Glc hederagenin,
hex-hex-bayogenin, 3-Glc-Glc-medicagenic acid, 3-Rha-
Gal-Glc-soyasapongenol B (soyasapogenol I), 3-Glc-
Ara-28-Glc-hederagenin, 3-Glc-medicagenic acid,3-Glc-
malonyl-medicageinc acid, Rha-Gal-GlcA soyasapogenol
E, Rha-hex-hex-soyasapogenol E, Ara-hex-hederagenin,
pen-hederagenin (Huhman & Sumner, 2002), zahnic
acids, soyasapogenol A, soyasapogenol B, soyasapogenol
E, bayogenin glycoside, 3-O-[-d-glucopyranosyl(13)-
-d-glucopyranosyl]-28-O--d-glucopyranoside
medicagenate, 3-O-[-l-rhamnopyranosyl(12)--d-
2011 Informa Healthcare USA, Inc.
Medicago sativa: A review 213
glucopyranosyl(1 2)- -d-glucopyranoside]
medicagenic acid, 3-O-[-d-glucopyranosyl(12)--d-
glucopyranosyl(12)--d-g lucopyranosyl]-28--d-
glucopyranoside medicagenate, 3-O-[-d-glucuronopy-
ranosylmethylester]-28-O-[-d-xylopyranosyl(14)--l-
rhamnopyranosyl(12)--l-arabinopyranoside]
medicagenate, 3-O-[-l-rhamnopyranosyl(12)--d-
galactopyranosyl(12)--d-glucuronopyranosyl]-21-
O--l-rhamnopyranoside soyasapogenol A, 3-O-[-d-
glucopyranosyl(12)--d-glucopyranosyl(12)
glucopyranosyl]-28-O-[-d-xylopyranosyl(14)--l-
rhamnopyranosyl(12)--l-arabinopyranoside] medica-
genate,3-O-[-d-glucopyranosyl(12)--d-glucopyranosyl
(1 2)glucopyranosyl]-28-O-{ -d-xylopyranosyl
(1 4)-[ -d-apiofuranosyl-(13)]- -l-rhamno
pyranosyl(12)--l-arabinopyranoside} medicagenate,
3-O-[-d-glucopyranosyl(12)--d-glucopyranosyl(12)--
d-glucopyranosyl]-28-O-[-d-xylopyranosyl(14)--l-
rhamnopyranosyl(12)--l-arabinopyranoside]zahnic
acid, 3-O-[-d-glucopyranosyl(12)--d-glucopyranosyl
(12)--d-glucopyranosyl]-28-O-{-d-xylopyra
nosyl(14)-[-d-apiofuranoside-(13)]--l-rhamno
pyranosyl(12)--l-arabinopyranoside}zahnic acid, 3-
O-[-d-galactopyranosyl(12)--d-glucuronopyranosyl]-
28-O--d-glucopyranoside bayogenin, 3-O-[-l-rham-
nopyranosyl(12)--d-galactopyranosyl(12)--d-
glucuronopyranoside] soyasapogenol E (Bialyetal., 1999),
3-O-[-d-glucopyranosyl (1-2)--d-glucopyranosyl(1-2)-
-d-glucopyranosyl]-2,3,16-trihydroxyolean-12-
en-23,28-dioic acid-23-O--l-arabinopyranosyl-28-O-
[-d-apiofuranosyl(1-3)--d-xylopyranosyl(14)-
-l-rhamnopyranosyl(1-2)- -l-arabinoside]
(Oleszek et al., 1992), 28-O-[-d-xylopyranosyl-(14)-
-l-rhamnopyranosyl(12)--l-arabinopyranosyl]
medicagenate,28-O-[-d-xylopyranosyl(14)--l-rham-
nopyranosyl(12)--l-arabinopyranosyl]-3-O--d-
glucopyranosylmedicagenate, 28-O-[-d-xylopyranosyl
(1 4)- -l-rhamnopyranosyl(1 2)- -l-
arabinopyranosyl]-3-O-[-d-glucopyranosyl(12)--d-
glucopyranosyl]medicagenate, 28-O-[-d-xylopyranosyl
(1 4)- -l-rhamnopyranosyl(1 2)- -l-
arabinopyranosyl]-3-O--d-glucuronopyranosyl-
medicagenate, 3-O-[-l-rhamnopyranosyl(12)--d-
glucopyranosyl(12)--d-glucuropyranosyl]soy-
asapogenol B (Massiot et al., 1991), dehydrosoyasa-
poninI, soyasaponin I, azukisaponin II, azukisaponin V
(Kitagawa etal., 1988). Sterols: -spinasterol, -sitosterol,
stigmasterol, myrsellinol, scopoletin, esculetin (El-Khrisy
et al., 1994; Abdel-Hafez, 1993), dihydrospinasterol,
24-methylcholest-7-enol, stigmasterol, campesterol
(Li-Shar & Claus, 1988). Thyrotropin-releasing hormone
analog (Worthington & Breskin, 1983). Vitamins: A, B1,
B6, B12, C, D, E, K, niacin, pantothenic acid, biotin, folic
acid (Worthington & Breskin, 1983; Horst et al., 1984).
Volatile components: terpenes, limonene, linalool, trans-
ocimene, furanoids, nonadienal, 2-methyl 4-pentenai,
benzaldehyde, ethyl benzaldehyde, alcohols, butanol,
hexanol, octanol, pentan-3-ol, 3-methylbutanol, trans-
214 Kundan Singh Bora, and Anupam Sharma
2-pentenol, trans-2-hexenol, trans-3-hexenol, pent-1-
en-3-ol, ott-I-en-3-ol, octa-1,5-dien-3-ol, benzyl alcohol,
2-phenylethanol, ketones, pent-1-en-3-one, pentan-3-
one, octan-3-one, methyl phenyl ketone, esters, trans-3-
hexenylacetate, trans-3-hexenylbutanoate, aldehydes,
hexanal, trans-2-pentenal, trans-2-hexenal, trans-2-
nonenal, trans-2,4-hexadienal, furane-2-ethyl (Aldo &
Luciano, 1997).
Figure 1 shows the chemical structures of common
saponin aglycones present in M. sativa.
Pharmacological/clinical reports
Several studies indicate that the ingestion of M. sativa
reduces cholesterol absorption and atherosclerotic
plaque formation in animals (Wilcox & Galloway, 1961;
Malinow et al., 1977a, 1981a; Cohen et al., 1990). M.
sativa top (stem and leaves) saponins have been reported
to decrease plasma cholesterol concentrations without
changing high-density lipoprotein cholesterol concen-
trations, decrease intestinal absorption of cholesterol,
increase excretion of neutral steroids and bile acids, pre-
vent atherosclerosis and induce the regression of athero-
sclerosis (Malinow et al., 1982a). Hypocholesterolemic
activity has been reported for root saponins, when given
to monkeys receiving a high-cholesterol diet (Malinow
etal., 1977b)
In a study, the ability of M. sativa plant to reduce
liver cholesterol accumulation in cholesterol-fed rats
was enhanced by the removal of saponins. Therefore,
M. sativa saponins appear to play an important role
in neutral steroid excretion, but are not essential for
increasing bile acid excretion (Story et al., 1984). In an
experiment with prairie dogs, the lowest incidence of
cholesterol gallstones was served with the diet of the
higher fiber content (85% alfalfa) (Cohen etal., 1990).
In a short-term study involving three normolipidemic
individuals given M. sativa seeds (80-60g daily), serum
cholesterol concentrations were reported to be reduced.
In another small study in which heat-treated M. sativa
seeds (40g three times daily for eight weeks) were taken
by eight type-IIA hyperlipoproteinemic patients and
three type IIB patients, a significant decrease was noted
in total serum cholesterol concentrations, low-density
lipoprotein (LDL) cholesterol and apolipoprotein B. The
LDL cholesterol concentration fell by less than 5% in two
of the 11 patients (Molgaard et al., 1987). Cholestaid,
a product available in the USA containing 900mg of
M.sativa extract with 100mg citric acid, is said to neu-
tralize the cholesterol in the stomach before it reaches
the liver, thus facilitating the excretion of cholesterol
from the body with no side effects or toxicity (Levy, 1999;
Dewey, 2001).
M. sativa top saponins have been shown to decrease
cholesterolemia without changing the levels of high
density lipoprotein-cholesterol; hence, they reduced the
total cholesterol/high density lipoprotein-cholesterol
ratio in Macaca fascicularis. Furthermore, they decreased
 Pharmaceutical Biology
intestinal absorption of cholesterol, increased fecal excre-
tion of endogenous and exogenous neutral steroids and
bile acids, and decreased the percentage distribution of
fecal deoxycholic and lithocholic acids (Malinow et al.,
1981a).
Khaleel etal. (2005) performed a study and showed that
highest saponin content extract just before fruiting stage
(free from both coumestrol and canavanine) of M. sativa
exhibited significant hypocholesterolemic and antiath-
erosclerotic activity. This study proved that M. sativa was
found to safely reduce natural cholesterol and to possess
a strong anti-atherosclerotic activity. The extracts pro-
duced the most significant decrease in total cholesterol
and LDL-cholesterol by 85.1 and 88%, respectively, of the
corresponding levels in hypercholesterolemic rabbits.
This decrease was more significant than that produced
by gemfibrozil (73 and 74%) upon concomitant admin-
istration with a cholesterol enriched diet using the same
animal model at the tested dose level. Moreover, it was
also observed that all M. sativa preparations produced
significant antioxidant properties.
M. sativa when supplied in the diet (6.25% by weight)
and infusion (1g/400mL) has been reported to reduce
the level of hyperglycemia in streptozotocin-induced
diabetes (Swanston etal., 1990). Aqueous extract of the
plant (1mg/mL) stimulated 2-deoxy-glucose transport
(1.8-fold), glucose oxidation (1.7-fold) and incorporation
of glucose into glycogen (1.6-fold) in mouse abdominal
muscle. In acute, 20min tests, 0.25-1mg/mL aque-
ous extract of M. sativa evoked a stepwise 2.5-6.3-fold
stimulation of insulin secretion from the BRIN-BD11
pancreatic beta cell line. This effect was abolished by
0.5mM diazoxide, and prior exposure to the extract did
not affect subsequent stimulation of insulin secretion by
10mM l-alanine, thereby negating a detrimental effect
on cell viability. The effect of the extract was potenti-
ated by 16.7mM glucose and by 1mM 3-isobutyl-1-
methylxanthine. l-Alanine (10mM) and a depolarizing
concentration of KCl (25mM) did not increase the insu-
lin-releasing activity of M. sativa. Sequential extraction
with solvents revealed insulin-releasing activity in both
the methanol and water fractions, indicating a cumula-
tive effect of more than one constituent (Mohamed etal.,
2006).
The manganese content of M. sativa (45.5mg/kg)
is reported to be the active principle responsible for a
hypoglycemic effect documented for M. sativa. A diabetic
patient, treated with soluble insulin but poorly controlled,
found that an M. sativa extract adequately controlled his
diabetes. When administered separately, only small doses
of manganese chloride (5-10mg) were required to have a
hypoglycemic effect. However, no effect was seen on the
blood sugar concentrations of non-diabetic controls or
of other diabetic patients, who were also administered
manganese. It was concluded that manganese lowered
the blood sugar concentration in this particular diabetic
patient because he was unable to utilize manganese
stored in his body (Rubenstein etal., 1962).
 Medicago sativa: A review 215

OH O CO2H
HO

HO HO HO
CH2OH
CH2OH CO2H

Soyasapogenol B Soyasapogenol E Medicagenic acid

OH
H

OH
HO COOH COOH

HO HO HO
CH2OH CH2OH CH2OH

Bayogenin Hederagenin Soyasapogenol A

OH O

OR2 CO2H
HO

O OH
R 1O R1 = GlcAP Galp Rhap OCH3
CH2OH R2 = H HO
CO2H
Soyasaponin I Formononetin Zahnic acid

Figure 1. Chemical structures of some common saponin aglycones present in M. sativa.

Methanol extract of M. sativa has been shown sig-
nificant estrogenic activity using an estrogen-dependent
MCF-7 breast cancer cell proliferation assay. The extract
showed significant competitive binding to estrogen
receptor (ER). The pure estrogen antagonist, ICI 182,780,
suppressed cell proliferation induced by the extract, sug-
gesting an ER-related signaling pathway was involved. The
ER subtype-selective activities of extract were examined
using transiently transfected human embryonic kidney
(HEK 293) cells. Methanol extract exhibited preferential
agonist activity toward ER. Phytoestrogens of the extract
were determined to be responsible for estrogenic activity
(Boue etal., 2003).
Extract of the leaves from M. sativa has been shown
to be used in treatment of neurovegetative menopausal
symptoms in women. Hot flushes and night sweating
completely disappeared with the treatment of M.sativa
extract. The plant product induced a significant increase
in prolactin and thyroid stimulating hormone response
to thyroid releasing hormone. Basal levels of estradiol,
luteinizing hormone, follicle stimulating hormone,
2011 Informa Healthcare USA, Inc.
prolactin, and thyroid stimulating hormone were
unchanged. Thus, M. sativa suggested having a cen-
tral slight antidopaminergic action without side effects
(Minerva, 1998).
Seed extract of M. sativa has been reported to exhibit
significant reduction of total cholesterol, phospholipid,
triglyceride, LDL-cholesterol and VLDL-cholesterol in
chicks, where clofibrate was used as a standard drug
(Dixit & Joshi, 1985).
In an in vitro experiment, polysaccharides isolated
from M. sativa exhibited immunopotentiating activity by
increasing mouse lymphocyte uptake of [3H] thymidine
(Zhao etal., 1993).
The antimicrobial activity of saponins isolated from
M. sativa against selected medically important yeasts,
Gram-positive and -negative bacteria has been investi-
gated. Increasing antibiotic activity was observed going
from the saponin extracts to the sapogenin samples,
suggesting that the sugar moiety is not important for
the antimicrobial efficacy. Activity was found especially
high against Gram-positive bacteria (Bacillus cereus,
216 Kundan Singh Bora, and Anupam Sharma
B. subtilis, Staphylococcus aureus, and Enterococcus
faecalis). Discrete antifungal activity was also observed,
mainly against Saccharomyces cerevisiae. The observed
antimicrobial properties of M. sativa were related to the
content of medicagenic acid (Avato etal., 2006).
An investigation into the effect of various herbs on
hepatic drug metabolizing enzymes in the rat, showed
that M. sativa potentiated the activity of aminopy-
rine N-demethylase but had no effect on glutathione
S-transferase or epoxide hydrolase activities (Garrett
etal., 1982).
M. sativa can be used as an alternative natural carote-
noid instead of the synthetic apo-ester in goldfish diets
(Yanar etal., 2008).
Medicagenic acid isolated from M. sativa exhibited
significant nematicidal activity against the plant-parasitic
nematode Xiphinema index (Argentieri etal., 2008).
Refined components of M. sativa polysaccharides
have been shown to inhibit the activities of reverse
transcriptase of HIV and protease of HIV (Zhang et al.,
2006).
Health beverage manufactured from M. sativa buds
was found beneficial in maintaining normal digestive
function and nutrition balance in human body, reducing
cholesterol, and preventing osteoporosis, arteriosclerosis
and aging (Song & Wei, 2007).
The saponin extract of M. sativa containing hederagen-
ins and saponin compounds I, II and III was reported to
be used in manufacturing of hypolipidemic agents and
antiatherosclerotics (Yu etal., 2007).
The dried powder of M. sativa has been shown to
inhibit the germination and growth of Echinochloa
crus-galli Wight (Gwon & Kim, 2006).
Xiong (2003) demonstrated that extracts prepared
from M. sativa roots may be used to prepare medical
preparations like powder, pill, or decoction for lowering
the levels of cholesterol and lipid in blood, improving the
liver function and the control and transmission of nerve
tissue, and treating calculus.
Among M. sativa secondary metabolites, saponins and
phytoestrogens offer interesting medicinal and nutraceu-
tical prospects. Phytoestrogens, mainly coumestrol, api-
genin and quercetin exhibit strong estrogenic activity and
have potential for use in treatment of hormone-related
cancers (Huyghe etal., 2007).
The extracts prepared from M. sativa flowery aerial
parts and seeds exhibited significant in vitro and in vivo
antimitotic activities (Dutu etal., 2002).
Saponins isolated from the aerial parts of M. sativa
have been shown to stimulate lipolytic activity without
influencing amylolytic or proteolytic activity of Neo-
Pancreatinum (Sroka etal., 1997).
Triterpenoid glycosides of the roots and leaves of
M. sativa have shown antibacterial activity against
Corynebacterium michiganense, Corynebacterium insi-
diosum, and Agrobacterium tumefaciens. The inhibitory
effects of these glycosides on cotton and cabbage seed
germinations have also been tested (Timbekova, 1995).
 Pharmaceutical Biology
Recently, the authors have shown that M. sativa exhib-
ited significant antioxidant and cerebroprotective effects
against ischemia and reperfusion insult in mice (Kundan
& Anupam, 2010). Pretreatment with M. sativa extract
(100 or 200mg kg1, p.o.) markedly reduced cerebral
infarct size, xanthine oxidase, superoxide anion and thio-
barbituric acid-reactive substance levels, significantly
restored reduced glutathione, superoxide dismutase and
total tissue sulfhydryl levels and attenuated impairment
in short-term memory and motor coordination. In addi-
tion, M. sativa directly scavenged free radicals generated
against a stable radical 1,1-diphenyl-2-picrylhydrazyl
and, superoxide anion radicals generated in phenazine
methosulfate-nicotinamide adenine dinucleotide sys-
tems, and also inhibited xanthine dehydrogenase/xan-
thine oxidase conversion and resultant superoxide anion
production.
In addition to the above mentioned activities, M. sativa
is also considered beneficial in arthritis (Hall, 1981), car-
diovascular complaints (Reilly, 1989), convalescence,
debility (Mills, 1994) and dysuria (Duke, 2002). The plant
is said to promote weight gain (Meyer, 1954), and can be
useful for people with protein allergies (Hall, 1981), GI
cramping and colic (Reilly, 1989), gravel (Duke, 2002),
hyperacidic stomach (Hall, 1981), hypoestrogenism/
hyperestrogenism (Reilly, 1989). It also improves appe-
tite (Lust, 1974), and intellect (Duke, 2002). l-Canaverine
isolated from the plant has been shown to have antitu-
mor activity against certain types of leukemia cells in
mice and selective toxicity in dog cancer cells grown
invitro (Hendler, 1995). M. sativa has been reported to be
beneficial in the treatment of peptic ulcers (Lust, 1974),
polycystic ovaries (Reilly, 1989), prolactin excess (Reilly,
1989), hemorrhage, as a tonic after blood loss and during
anemia (Mills, 1994), urinary and bowel problems (Lust,
1974), scurvy (Duke, 2002), secondary hypothyroidism
(Reilly, 1989) and Herpes simplex (Hendler, 1995).
Toxicity/side-effects
Both M. sativa seed and herb have been reported to
induce a systemic lupus erythematosus (SLE)-like syn-
drome in female monkeys (Barnes et al. 2007; Boon &
Smith, 2004; Brinker, 2001; Malinow etal., 1982a, 1982b).
This activity has been attributed to canavanine, a non-
protein amino acid constituent, which has been found
to have effects on human immunoregulatory cells in
vitro (Jorge etal., 1985). Re-activation of quiescent SLE
in humans has been associated with the ingestion of M.
sativa tablets which, following analysis, were found to
contain canavanine (Roberts & Hayashi, 1983). It was
not stated whether the tablets contained seed or herb
material. Canavanine is known to be toxic to all animal
species because it is a structural analogue of arginine
and may interfere with the binding of this amino acid to
enzymes and its incorporation into proteins. M. sativa
seeds are reported to contain substantial quantities of
canavanine (8.33-13.6mg/kg), whereas the herb is stated

to contain amounts that are considerably less (Natelson,
1985).
Pancytopenia has been associated with human inges-
tion of ground M. sativa seeds (80-160g/day), which
were taken to lower plasma cholesterol concentrations
(Malinow etal., 1981a). Dietary studies using alfalfa top
saponins (ATS) in the diet of rats and monkeys showed
no evidence of toxicity and serum lipid concentrations
were lowered (Malinow etal., 1982a, 1981a). In addition,
when ATS were given to cholesterol-fed animals, a reduc-
tion in serum lipid concentrations was observed. ATS are
reported to be free of the SLE-inducing substance that
is present in the seeds (Malinow etal., 1982c). Negative
results were documented for M. sativa when tested for
mutagenicity using Salmonella strains TA98 and TA100
(White etal., 1983).
Consultation is a necessity for a health care practi-
tioner prior to prescribe use of M. sativa if undergoing
hormone replacement therapy or taking birth control
medication or using blood thinners (Barnes etal. 2007;
Boon & Smith, 2004; Brinker, 2001).
Doses
Preparations equivalent to 930g dried aerial parts,
per day (Mills, 1985; BHMA, 1983)
Dried aerial parts: 510g, 3 times per day (BHMA,
1983). 310g, 3 times per day (Mills, 1985)
Infusion: 510g dried aerial parts, 3 times per day
(BHMA, 1983)
Fluid extract: 510g dried equivalent, 3 times per day
(BHMA, 1983) (1:1, 25% alcohol, 510mL)
Tablets: A dose of two tablets (1g each) of Cholestaid
(esterin processed M. sativa) taken by mouth 3 times
daily for up to two months, then one tablet 3 times
daily, has been recommended (Farnsworth, 1995).
Seeds: For treating high cholesterol, a dose of 40g of
heated seeds taken by mouth 3 times daily has been
recommended (Farnsworth, 1995).
Contraindications
People with a history of lupus or a family history of
systemic lupus erythematosus should avoid M. sativa
supplements (Barnes etal. 2007; Boon & Smith, 2004;
Brinker, 2001).
M. sativa seeds should not be ingested during preg-
nancy or lactation (Brinker, 2001).
Discussion
Plants have played a significant role in maintaining
human health and improving the quality of human life
for thousands of years, and have served humans well as
valuable components of medicines, seasonings, bev-
erages, cosmetics and dyes. Herbal medicine is based
on the premise that plants contain natural substances
that can promote health and alleviate illness. In recent
times, focus on plant research has increased all over the
world and a large body of evidence has collected to show
2011 Informa Healthcare USA, Inc.
Medicago sativa: A review 217
immense potential of medicinal plants used in various
traditional systems. The present review emphasizes the
phytochemical, traditional, pharmacological, clinical
and toxicological reports on M. sativa. Saponins, fla-
vonoids, phytoestrogens, coumarins, alkaloids, amino
acids, phytosterols, vitamins, digestive enzymes and
terpenes constitute major classes of phytoconstituents
of the plant. M. sativa has been used since centuries as
homoeopathic and Ayurvedic medicine for a variety of
ailments. The plant is widely used in foods and is listed
by the Council of Europe as a source of natural food fla-
vor (category N2 and N3). These categories indicate that
M.sativa can be added to foodstuffs in small quantities
with a limitation on the concentrations of an active
ingredient in the final product. In the USA, M. sativa is
listed as GRAS (generally recognized as safe). Recent
research carried out indicates its uses such as neuro-
protective, hypocholesterolemic, antioxidant, antiulcer,
antimicrobial, hypolipidemic, immunopotentiating,
estrogenic, digestive, nutritive for human body, and in
the treatment of atherosclerosis, heart disease, stroke,
cancer, diabetes and neurovegetative menopausal
symptoms in women.
The presence of a wide range of chemical com-
pounds indicates that the plant could lead the way for
the development of novel agents having good biologi-
cal activity. Exploration of the chemical compounds
of the plant will provide the basis for developing such
a lead. Many chemical compounds are present in the
plant but isolation of active constituents by using vari-
ous appropriate chromatographic techniques should
be carried out. Further studies can be carried out
using different extraction methods such as microwave
extraction, isolation and identification of active com-
pounds, pharmacological screening of extracts as well
as isolated compound(s) so that accuracy of results can
be obtained and use of extracts can be made in differ-
ent herbal formulation. Despite a long tradition of use
of M. sativa for treatment of various ailments, only spo-
radic pharmacological work has so far been carried out
to prove its traditional claims. Additionally, the plant
has been included in a number of herbal and homoeo-
pathic formulations, which are in clinical use for the
treatment of various ailments. M. sativa seems to hold
great potential for in depth investigation for various
biological activities. Therefore, it is necessary to exploit
its maximum potential in the field of medicinal and
pharmaceutical sciences for novel and fruitful applica-
tion Currently, the authors are involved in evaluating
the CNS effects of traditionally used medicinal plants,
including M. sativa, with a view to isolating bioactive
phytoconstituent(s).
Declaration of interest
Financial support from the L.L.R. Educational Trust,
Solan, Himachal Pradesh, India, which runs the
L.R. Institute of Pharmacy, is gratefully acknowledged.
218 Kundan Singh Bora, and Anupam Sharma
The authors alone are responsible for the content and
writing of the paper.
References
Abdel-Hafez OM. (1993). Constituents of Medicago sativa fruits.
Fitoterapia, 64, 381382.
Adler J. (1962). Anti-oestrogenic activity in alfalfa. Vet Record, 74,
11481154.
Albourine A, Petit-Ramel M, Thomas-David G, Vallon JJ. (1991).
Extraction of l-canavanine from soybean (Glycine max) and
alfalfa (Medicago sativa). Analysis by high-performance liquid
chromatography. J Pharm Belg, 46, 229235.
Aldo T, Luciano P. (1997). Volatiles from Medicago sativa complex
flowers. Phytochemistry, 45, 11451148.
Argentieri MP, DAddabbo T, Tava A, Agostinelli A, Jurzysta M, AvatoP.
2008. Evaluation of nematicidal properties of saponins from
Medicago spp. Eur J Plant Pathol, 120, 189197.
Avato P, Bucci R, Tava A, Vitali C, Rosato A, Bialy Z, Jurzysta M. (2006).
Antimicrobial activity of saponins from Medicago spp. structure-
activity relationship. Phytother Res, 20, 454457.
Barcelo F, Mirales F, Arean CO. (1997). Purification and characterization
of ferritin from alfalfa seeds. J Inorg Biochem, 66, 2327.
Barnes DK, Sheaffer CC. (1995). Alfalfa. In: Barnes RF, Miller DA,
Nelson CJ, eds. Forages: An Introduction to Grassland. Ames: Iowa
State University, pp.205216.
Barnes J, Anderson LA, Philipson JD. (2007). Herbal Medicines, third
edition. London: Pharmaceutical Press.
Bhavna G, Rashmi SS, Radha G. (2007). Therapeutic uses of Euphorbia
thymifolia: A Review. Phcog Rev, 1, 299304.
BHMA (1983). British Herbal Pharmacopoeia (BHP). Cowling
(Yorkshire): British Herbal Medical Association.
BHMA (1996). British Herbal Pharmacopoeia (BHP). Bournemouth:
British Herbal Medical Association.
Biagioni M, Alpi A, Picciarelli P. (1990). Aminopeptidases (EC.3.4.11)
in alfalfa (Medicago sativa L.) leaves. J Plant Physiol, 135, 559564.
Bialy Z, Jurzysta M, Oleszek W, Piacente S, Pizza C. (1999). Saponins
in alfalfa (Medicago sativa L.) root and their structural elucidation.
JAgr Food Chem, 47, 31853192.
Bickhoff EM, Livingston AL, Booth AN. (1960). Estrogenic activity of
coumestrol and related compounds. Arch Biochem Biophys, 88,
262267.
Bickhoff EM, Livingston AL, Booth AN. (1964). Tricin from alfalfa-
isolation and physiologic activity. J Pharm Sci, 53, 1411.
Boon H, Smith MJ. (2004). The Complete Natural Medicine Guide to the
50 Most Common Medicinal Herbs, second edition. Toronto (ON):
Robert Rose.
Boue SM, Wiese TE, Nehls S, Burow ME, Elliott S, Carter-WientjesCH,
Shih BY, McLachlan JA, Cleveland TE. 2003. Evaluation of the
estrogenic effects of legume extracts containing phytoestrogens.
JAgr Food Chem, 51, 21932199.
Brinker F. (2001). Herb Contraindications and Drug Interactions, third
edition. Sandy, OR: Eclectic Medical.
Cohen BI, Mosbach EH, Matoba N, Suh SO, McSherry CK. (1990).
The effect of alfalfa-corn diets on cholesterol metabolism and
gallstones in prairie dogs. Lipids, 25, 143148.
DerMarderosian A, Liberti L, Beutler JA, Grauds C, Tatro DS,
Cirigliano M, DeSilva D Jr. (2005). Alfalfa. Review of Natural
Products: Facts and comparisons. Alphen aan den Rijn, the
Netherlands: Wolters Kluwer Health.
Dewey D. (2001). Cholestaid. NuPharma, January 1. http://www.
drugs.com/npp/alfalfa.html (accessed 15 January 2010).
Dixit VP, Joshi SC. (1985). Antiatherosclerotic effects of alfalfa meal
ingestion in chicks: A biochemical evaluation. Indian J Physiol
Pharmacol, 29, 4750.
Dixon RA, Steele CL. (1999). Flavonoids and isoflavonoids A gold
mine for metabolic engineering. Trends Plant Sci, 4, 394400.
Doehlert DC, Duke SH, Anderson L. (1982). Amylases from alfalfa
(Medicago sativa L.) roots. Plant Physiol, 69, 10961102.
 Pharmaceutical Biology
Duke J. (1985). Handbook of Medicinal Herbs. Boca Raton, FL: CRC
Press.
Duke J. (2002). American Agricultural Research Service-Phytochemical
& Ethnobotanical Database. Available at: http://www.ars-grin.
gov/duke/index.html (accessed 10 August 2009).
Dutu LE, Istudor V, Loloiu T, Radulescu V. (2002). Research on
polyphenolic compounds from Medicago sativa L. Farmacia
(Bucharest Romania), 50, 4456.
Ehsanpour AA, Razavizadeh R. (2005). Effect of UV-C on drought
tolerance of alfalfa (Medicago sativa) callus. Am J Biochem
Biotechnol, 1, 107110.
El-Khrisy EAM, Abdel Hafez OM, Khattab AA, Ahmed KM. (1994).
Chemical constituents of Medicago sativa L. Bull Nat Res Cent
(Egypt), 19, 117122.
FAO. (2006). FAOSTATFood and Agriculture Organization of the
United Nations. Available at http://faostat.fao.org (accessed 05
March 2010).
Farnsworth NR. (1995). Alfalfa pills and autoimmune diseases. Am
JClin Nutr, 62, 10261028.
Finkler K. (1985). Spiritualist Healers in Mexico. South Hadley, MA:
Bergin & Garvey.
Francoise F, Daniel Le R, John GS. (1991). Effects of salt stress on
amino acid organic acid and carbohydrate composition of roots
bacteroids and cytosol of Alfalfa (Medicago sativa L.). Plant Physiol,
96, 12281236.
Fushiya S, Tamura T, Tashiro T, Nozoe S. (1984). Structure and
synthesis of medicanine, a new amino acid from Medicago sativa.
Heterocycles, 22, 10391040.
Garrett BJ, Cheeke PR, Miranda CL, Goeger DE, Buhler DR. (1982).
Consumption of poisonous plants (Senecio jacobaea, Symphytum
officinale, Pteridium aquilinum, Hypericum perforatum) by
rats chronic toxicity mineral metabolism and hepatic drug-
metabolizing enzymes. Toxicol Lett, 10, 183188.
Gray AM, Flatt PR. (1997a). Pancreatic and extra-pancreatic effects of
the traditional anti-diabetic plant Medicago sativa (Lucerne). Brit
J Nutr, 78, 325334.
Gray AM, Flatt PR. (1997b). Natures own pharmacy the diabetes
perspective. P Nutr Soc, 56, 507-5 I7.
Gupta BK, Nandra KS, Chopra AK. (1981). Chemical and mineral
composition on different cuttings of lucerne (Medicago sativa).
JRes (Punjab Agricultural University), 18, 345348.
Gwon CH, Kim JD. (2006). Plant ground powder or extract of M. sativa
L. Avena sativa L. or Cirsium japonicum having growth inhibition
effect of Echinochloa crusgalli Wight. Korean Republic Kongkae:
Taeho Kongbo.
Hall D. (1981). The Herb Tea Book. Connecticut: Keats Publishing,
234239.
Hendler SS. (1995). The Doctors Vitamin & Mineral Encyclopedia.
London: Leopard.
Heywood VH, Ball PW. (1968). Leguminosae. In: Tutin TG, Heywood
VH, Burges NA, Moore DM, Valentine DH, Walters SM, Webb DA,
Eds. Flora Europaea, Vol II. Cambridge: Cambridge University
Press, pp. 245260.
Horst RL, Reinhardt TA, Russell JR, Napoli JL. (1984). The isolation
and identification of vitamin D2 and vitamin D3 from Medicago
sativa (alfalfa plant). Arch Biochem Biophys, 231, 6771.
Huhman DV, Sumner LW. (2002). Metabolic profiling of saponins in
Medicago sativa and Medicago truncatula using HPLC coupled to
an electrospray ion-trap mass spectrometer. Phytochemistry, 59,
347360.
Huyghe C, Bertin E, Landry N. (2007). Medicinal and Nutraceutical
Uses of Alfalfa (Medicago sativa L.). A review. In: Acharya SN,
Thomas JE, eds. Advances in Medicinal Plant Research. Trivandrum,
Kerala, India: Research Signpost, pp. 147172.
Inamul H. (2004). Safety of medicinal plants. Pakistan J Med Res, 43, 18.
Jorge AV, Antonio I, Luis L, Donato AS. (1985). Effects of l-canavanine on
T cells may explain the induction of systemic lupus erythematosus
by alfalfa. Arthritis Rheum, 28, 5257.
Khaleel AEG, Mohamed Z, El-Maraghy SA, Hifnawy MS, Abdel-SattarE.
(2005). Study of hypocholesterolemic and antiatherosclerotic

properties of Medicago sativa L. cultivated in Egypt. Yaowu Shipin
Fenxi, 13, 212218.
Kitagawa I, Taniyama T, Murakami T, Yoshihara M, Yoshikawa M.
(1988). Saponin and sapogenol. XLVI. On the constituents in
aerial part of American alfalfa Medicago sativa L. The structure of
dehydrosoyasaponin I. Yakugaku Zasshi, 108, 547554.
Kowalska I, Stochmal A, Kapusta I, Janda B, Pizza C, Piacente S,
Oleszek W. (2007). Flavonoids from Barrel Medic (Medicago
truncatula) aerial parts. J Agr Food Chem, 55, 26452652.
Kundan SB, Anupam S. (2010). Evaluation of antioxidant and
cerebroprotective effect of Medicago sativa Linn. against ischemia
and reperfusion insult. eCAM, 19. Available at: http://ecam.
oxfordjournals.org (accessed 16 April 2010).
Levy S. (1999). New product newswire. Drug Top, 19, 2226.
Lihu M, Pedak E. (1979). Chemical composition of alfalfa (Medicago
sativa L.) in different stages of growth. Teaduslike Toode Kogumik,
49, 104109.
Li-Shar H, Claus G. (1988). Sterol and phospholipid changes during
alfalfa seed germination. Phytochemistry, 27, 20492053.
Lust J. (1974). The Herb Book. New York: Benedict Lust Publications.
Malinow MR, Bardana EJJ, Goodnight SHJ. (1981a). Pancytopenia
during ingestion of alfalfa seeds. Lancet, 1, 615.
Malinow MR, Bardana EJJ, Pirofsky B, Craig S, McLaughlin P. (1982b).
Systemic lupus erythematosus-like syndrome in monkeys fed alfalfa
sprouts role of a nonprotein amino acid. Science, 216, 415417.
Malinow MR, Connor WE, McLaughlin P, Stafford C, Lin DS,
LivingstonAL, Kohler GO, McNulty WP. (1981b). Cholesterol and
bile acid balance in Macaca fascicularis. Effects of alfalfa saponins.
J Clin Invest, 67, 156162.
Malinow MR, McLaughlin P, Papworth L, Stafford C, Kohler GO, Livingston
AL, Cheeke PR. (1977a). Effect of alfalfa saponins on intestinal
cholesterol absorption in rats. Am J Clin Nutr, 30, 20612067.
Malinow MR, McLaughlin KGO, Livingston AL. (1977b). Prevention of
elevated cholesterolemia in monkeys by alfalfa saponins. Steroids,
29, 105110.
Malinow MR, McNulty WP, Houghton DC, Kessler S, Stenzel P,
Goodnight SHJ, Bardana EJJ, Palotay JL, McLaughlin P,
Livingston AL. (1982a). Lack of toxicity of alfalfa saponins in
Cynomolgus macaques. J Medical Primatol, 11, 106118.
Malinow MR, McNulty WP, McLaughlin P, Stafford C, Burns AK,
Livingston AL, Kohler GO. (1981c). The toxicity of alfalfa saponins
in rats. Food Cosmet Toxicol, 19, 443445.
Massiot G, Catherine L, Dominique G, Louisette Le MO. (1988).
Reinvestigation of the sapogenins and prosapogenins from alfalfa
(Medicago sativa). J Agr Food Chem, 36, 902.
Massiot G, Lavaud C, Besson V, Le Men-Olivier L, Van Binst G. (1991).
Saponins from aerial parts of alfalfa (Medicago sativa). J Agr Food
Chem, 39, 7882.
Mego V, Erdelsky K. (1977). Free amino acids in alfalfa (Medicago
sativa) L. Physiol Plantarum, 13, 4955.
Meyer JH. (1954). The utilization of alfalfa and alfalfa fiber fractions by
growing rats. J Nutr, 237247.
Michael A, Francine G, Matthias H. (2007). Plants traditionally used in
age related brain disordersA survey of ethnobotanical literature.
JEthnopharmacol, 113, 363381.
Mills S. (1985). The Dictionary of Modern Herbalsim. Wellingborough:
Thorsons.
Mills S. (1994). The Complete Guide to Modern Herbalism. Wellingborough:
Thorsons.
Minerva G. (1998). Treatment of neurovegetative menopausal
symptoms with a phytotherapeutic agent. Universita degli Studi-
Siena, 50, 207211.
Mohamed B, Abderrahim Z, Hassane M, Abdelhafid T, Abdelkhaleq L.
(2006). Medicinal plants with potential antidiabetic activity A
review of ten years of herbal medicine research (19902000). Int J
Diab Metab, 14, 125.
Molgaard J, von Schenck H, Olsson AG. (1987). Alfalfa seeds lower
low density lipoprotein cholesterol and apolipoprotein B
concentrations in patients with type II hyperlipoproteinemia.
Atherosclerosis, 65, 173179.
2011 Informa Healthcare USA, Inc.
Medicago sativa: A review 219
Natelson S. (1985). Canavanine in alfalfa (Medicago sativa). Experientia,
41, 257259.
Natelson S. (1985). Canavanine to arginine ratio in alfalfa (Medicago
sativa) clover (Trifolium) and the jack bean (Canavalia ensiformis).
J Agr Food Chem, 33, 413419.
Newby VK, Sablon RM, Synge RLM, Vande CK, Van S, Christiaan F.
(1980). Free and bound phenolic acids of lucerne (Medicago sativa
cv Europe). Phytochemistry, 19, 651657.
Oleszek W. (1988). Solid-phase extraction-fractionation of alfalfa
saponins. J Sci Food Agr, 44, 4349.
Oleszek W, Jurzysta M. (1986). Isolation chemical characterization and
biological activity of alfalfa (Medicago media Pers.) root saponins.
Acta Soc Bot Pol, 55, 2333.
Oleszek W, Jurzysta M, Ploszynski M, Colquhoun IJ, Price KR,
FenwickGR. (1992). Zahnic acid tridesmoside and other dominant
saponins from alfalfa (Medicago sativa L.) aerial parts. J Agr Food
Chem, 40, 191196.
Orr JD, Sumner LW, Edwards R, Dixon RA. (1993). Determination of
cinnamic acid and 4-coumaric acid in alfalfa (Medicago sativa L.)
cell suspension cultures by gas chromatography. Phytochem Anal,
4, 124130.
Parry AD, Edwards R. (1994). Characterization of flavonoid
O-glycosyltransferases from alfalfa (Medicago sativa). Acta
Horticulturae, 381, 154157.
Piretti MV, Zeli F, Pistore R. (1982). Flavonoid polyphenols in alfalfa
(Medicago sativa). Gazz Chim Ital, 112, 4750.
Potgieter SJ. (2005). Nutritional supplement extracted from alfalfa
(Medicago sativa). PCT Int. Appl. WIPO Patent Application
WO/2005/002357 (South Africa) Available at http://www.wipo.int/
pctdb/en/wo.jsp?wo=2005002357 (accessed 02 March 2010).
Reilly P. (1989). Clinical application-Medicago sativa extracts.
JNaturopathic Med, 1, 455460.
Roberts JL, Hayashi JA. (1983). Exacerbation of SLE associated with
alfalfa ingestion. New Engl J Med, 308, 1361.
Rodriguez-Garay B, Phillips GC, Kuehn GD. (1989). Detection of
norspermidine and norspermine in Medicago sativa L. (alfalfa).
Plant Physiol, 89, 525529.
Rubenstein AH, Levin NW, Elliott GA. (1962). Manganese-induced
hypoglycemia. Lancet, 2, 13481351.
Shao H, Dixon RA, Wang X. (2007). Crystal structure of vestitone
reductase from alfalfa (Medicago sativa L). J Mol Biol, 369, 265276.
Song Y, Wei J. (2007). Health beverage containing Medicago sativa.
Faming Zhuanli Shenqing Gongkai Shuomingshu, 7.
Sroka Z, Jurzysta M, Tylcz J, Rzadkowska-Bodalska H. (1997).
Stimulation of pancreatic lipase activity by saponins isolated from
Medicago sativa. Z Naturforsch C, 52, 235239.
Steppler HA. (1987). ICRAF and a decade of agroforestry development.
In: Steppler HA, Nair PKR, eds. Agroforestry: A Decade of
Development. Nairobi, Kenya: International Council for Research
in Agroforestry, pp. 1324.
Stochmal A, Simonet AM, Macias FA, Oliveira MA, Abreu JM, Nash
R, Oleszek W. 2001. Acylated apigenin glycosides from alfalfa
(Medicago sativa L. var. Artal). Phytochemistry, 57, 12231226.
Stochmal A, Oleszek W. (2007). Seasonal and structural changes of
flavones in alfalfa (Medicago sativa) aerial parts. J Food Agr Env,
5, 170174.
Story JA, LePage SL, Petro MS, West LG, Cassidy MM, Lightfoot FG,
Vahouny GV. (1984). Interactions of alfalfa plant and sprout
saponins with cholesterol in vitro and in cholesterol-fed rats. Am
JClin Nutr, 39, 917929.
Sumner LW, Paiva NL, Dixon RA, Geno PW. (1996). High performance
liquid chromatography/continuous-flow liquid secondary ion
mass spectrometry of flavonoid glycosides in leguminous plant
extracts. J Mass Spectrom, 31, 472485.
Swanston SK, Day C, Bailey CJ, Flatt PR. (1990). Traditional plant
treatments for diabetes. Studies in normal and streptozotocin
diabetic mice. Diabetologia, 33, 462464.
Tamsyn SA, Thring PH, Declan PN. (2009). Anti-collagenase anti-
elastase and anti-oxidant activities of extracts from 21 plants. BMC
Complement Altern Med, 9, 2737.
220 Kundan Singh Bora, and Anupam Sharma
Teuber LR, Brick MA. (1988). Morphology and anatomy. In: HansonAA,
Barnes DK, Hill Jr RR, eds. Alfalfa and Alfalfa Improvement.
Madison, WI, 125162.
Timbekova AE. (1995). Chemistry and Biological Activity of Glycosides
from Medicago sativa. Book of Abstracts of the 210th ACS National
Meeting, Chicago, IL, August 20-24. Washington, DC: American
Chemical Society.
Timbekova AE, Isaev MI, Abubakirov NK. (1996). Chemistry and
biological activity of triterpenoid glycosides from Medicago sativa.
Adv Exp Med Biol, 405, 171182.
Toth EC, Vissi E, Kovacs I, Szoke A, Arino J, Gergely P, Dudits D,
Dombradi V. (2000). Protein phosphatase 2A holoenzyme
and its subunits from Medicago sativa. Plant Mol Biol, 43,
527536.
Volynets AP, Dubikovskii GP, Bardinov FG, Palchenko LA, Tikhon ZK.
(1979). Study of the phenol compounds of alfalfa Medicago sativa.
Seryya Biyalagichnykh Navuk, 4, 4044.
White RD, Krumperman PH, Cheeke PR, Buhler DR. (1983). An
evaluation of acetone extracts from six plants in the Ames
mutagenicity test. Toxicol Lett, 15, 2531.
Wieslaw O, Keith RP, Ian JC, Marian J, Michal P, Roger, GF. (1990).
Isolation and identification of alfalfa (Medicago sativa L.) root
saponins their activity in relation to a fungal bioassay. J Agr Food
Chem, 38, 18101817.
 Pharmaceutical Biology
Wilcox EB, Galloway LS. (1961). Serum and liver cholesterol total lipids
and lipid phosphorus levels of rats under various dietary regimes.
Am J Clin Nutr, 9, 236243.
Worthington RB, Breskin MF. (1983). A pharmacists guide to
controversial nutrition products Am Pharm, 23, 3042.
Xiaoqiang W, Xianzhi H, Jianqiao L, Hui S, Zhenzhan C, Richard
AD. (2006). Crystal structure of isoflavone reductase from alfalfa
(Medicago sativa L.). J Mol Biol, 358 13411352.
Xiong S. (2003). Medicinal-use extract of Medicago sativa root. Faming
Zhuanli Shenqing Gongkai Shuomingshu, 6.
Yanar M, Ercen Z, Hunt AO, Buyukcapar HM. (2008). The use of alfalfa
(Medicago sativa) as a natural carotenoid source in diets of goldfish
Carassius auratus. Aquaculture, 284, 196200.
Yu C, Guo J, Shen H, Liang D, Zhao N, Wen J. (2007). Method for
preparing saponin extract of Medicago sativa and its application
in manufacturing hypolipidemic agents and antiatherosclerotics.
Faming Zhuanli Shenqing Gongkai Shuomingshu, 10.
Zhang L, Zhang D, Feng K. (2006). Inhibition of refined components
of Medicago sativa polysaccharides to the activities of reverse
transcriptase of HIV and protease of HIV. Zhongguo Shipin Xuebao,
6, 5962.
Zhao WS, Zhang YQ, Ren LJ, Zhang L, Yang J. (1993).
Immunopotentiating effects of polysaccharides isolated from
Medicago sativa L. Zhongguo Yao Li Xue Bao, 14, 273276.