全 文 ： 2013 年 5 月 第 11 卷 第 3 期 Chin J Nat Med May 2013 Vol. 11 No. 3 199

Chinese Journal of Natural Medicines 2013, 11(3): 0199−0206
doi: 10.3724/SP.J.1009.2013.00199
Chinese
Journal of
Natural
Medicines







Hybanthus enneaspermus (L.) F. Muell: a concise
report on its phytopharmacological aspects
D. K. Patel, R. Kumar, K. Sairam, S. Hemalatha*
Pharmacognosy Research Laboratory, Department of Pharmaceutics, Institute of Technology, Banaras Hindu University, Varanasi 221005,
India
Available online 20 May 2013
[ABSTRACT] Hybanthus enneaspermus (L.) F. Muell belonging to the family Violaceae, popularly known as Ratanpurus (Hindi) is a
herb or a shrub distributed in the tropical and subtropical regions of the world. In the Ayurvedic literature, the plant is reported to cure
conditions of “Kapha” and “Pitta”, urinary calculi, strangury, painful dysentery, vomiting, burning sensation, wandering of the mind,
urethral discharge, blood trouble, asthma, epilepsy, cough, and to give tone to the breasts. Phytochemically, the plant contains a consid-
erable amount of dipeptide alkaloids, aurantiamide acetate, isoarborinol, and β-sitosterol, sugars, flavonoids, steroids, triterpenes, phe-
nols, flavones, catachins, tannins, anthraquinones and amino acids. Pharmacologically, the plant is reported to possess antidiabetic,
antiplasmodial, antimicrobial, anticonvulsant, nephroprotective, aphrodisiac, hepatoprotective, antiinflammatory, aldose reductase
inhibitory and free radical scavenging activities. The information provided in this review will be worthwhile to know the applicability
of H. enneaspermus for the treatment of various acute or chronic diseases with a diverse nature of phytoconstituents. The overall data
in this review article were collected from various scientific sources on the research of H. enneaspermus.
[KEY WORDS] Antidiabetic; Antimicrobial; Antioxidant; Aphrodisiac; Hybanthus enneaspermus; Pharmacological activity; Phytochemis-
try; Tissue culture
[CLC Number] R284 [Document code] A [Article ID] 1672-3651(2013)03-0199-08

1 Introduction
Hybanthus enneaspermus (L.) F. Muell. (H. enneaspermus),
belonging to the family Violaceae, is a herb or a shrub dis-
tributed in the tropical and subtropical regions of world, and
occurs mostly in the warmer parts of Deccan peninsula in
India. The plant is popularly called “Ratanpurus” by the local
Yanadi and Santal tribes, villagers and herbalists. This eth-
nobotanical herb is known to have unique medicinal proper-
ties [1]. It is also known as “hump back flower”, and is a
member of a genus of perennial herbs, often creeping, whose
leaves are alternate or in clusters. The genus consists of 150
species found in different regions of the world, often seen in
mountainous regions [2]. In Ayurveda, Siddha, and other tra-
ditional systems of medicine H. enneaspermus is used to treat
various ailments. In Ayurveda, it is known as Sthalakamala

[Received on] 08-May-2012
[Research funding] This project was supported by the financial assis-
tance from University Grants Commission, New Delhi.
[*Corresponding author] S. Hemalatha: Prof., E-mail: she-
malatha.phe@itbhu.ac.in, Tel: 91-9415256481
These authors have no conflict of interest to declare.
and is reported to have diuretic and demulcent properties.
The roots are used as a diuretic and in the treatment of gon-
orrhea, urinary infections, and in the case of cholera, in the
form of infusion. It is very effective in the treatment of in-
fectious diseases, and is said to have antidiabetic, tonic, diu-
retic, aphrodisiac and demulcent properties. Pharmacologi-
cally, the plant has been reported to have antidiabetic, an-
tiplasmodial, antimicrobial, anticonvulsant, nephroprotective,
aphrodisiac, hepatoprotective, antiinflammatory, aldose re-
ductase inhibitory, and free radical scavenging activities. The
natural regeneration potential of H. enneaspermus is very
poor due to the low seed viability [1].
2 Geographical Distribution and Classification
H. enneaspermus is derived from a Greek word hypos
which means humpback flower, referring to the drooping
pedicels of plants that are a part of this genus. It is an impor-
tant plant in the Indian system of medicine, and is popularly
known as Lakshmisheshta, Padmavati, Padmacharini or Pu-
rusharathna in Sanskrit [3]. It is a small suffrutescent perennial
herb found mostly in the tropical and subtropical regions of
the world particularly in places like tropical Asia, Africa,
Australia, Arabia, Sri Lanka, Indochina, Philippines, Borneo,
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200 Chin J Nat Med May 2013 Vol. 11 No. 3 2013 年 5 月 第 11 卷 第 3 期

Java, Guinea, Taiwan, and in India [4]. The genus Hybanthus
Jacq. (Violaceae) comprises of over 100 species, which are
mostly herbs, shrubs, and treelets distributed widely
throughout the world. Almost 70 species are distributed
mostly in Latin America, 20 species in Africa and Madagas-
car, 5 in Asia, and 11 in Australia [5]. In India, the genus is
mainly found in warmer parts of the Deccan peninsula and
grows along with many other simulating weeds viz. Ammania
baccifera, Oldenlandia alata, Heliotropium bracteatum and
Lindernia oppositifolia. It is pubescent in nature and grows
up to a height of 15−30 cm, and forms many diffuse or as-
cending branches. It has been noticed that the flowering sea-
son of Hybanthus occurs from August to September, and all
taxa of the Violaceae occur at high altitude [6]. Taxonomically,
it is classified and placed into the Kingdom: Plantae, Division:
Magnoliophyta, Class: Magnoliopsida, Order: Malpighiales,
Family: Violaceae, Genus: Hybanthus, Species: Hybanthus
enneaspermus. It is known by various names in different
language, such as in English: Spade Flower, Pink ladies slip-
per, in Hindi: Ratan purush, in Bengali: Munbora, in Kan-
nada: Purusharathna, in Malayalam and Tamil: Orilathamartai,
in Telugu: Ratnapurusha, and in Marathi: Rathanparas.
3 Traditional Uses
H. enneaspermus was found to be well utilized in the
treatment of various simple and complicated disease and
disorders. Many indigenous groups in India found their way
of utilizing it in various forms to treat different ailments. In
the Ayurvedic literature, the plant is reported to cure condi-
tions of “Kapha” and “Pitta”, urinary calculi, strangury,
painful dysentery, vomiting, burning sensation, wandering of
the mind, urethral discharge, blood trouble, asthma, epilepsy,
cough, and to give tone to the breasts [7]. Preparations made
from the leaves and tender stalks of H. enneaspermus are
used in herbal medicine for their aphrodisiac, demulcent, and
tonic properties. The roots are used as a diuretic and adminis-
trated as an infusion in gonorrhea and urinary infections. The
fruits and leaves are used as antidotes for scorpion stings and
cobra bites by the Yanadi tribes [1]. The leaves and tender
stalks are demulcent and used as a decoction or electuary;
mixed with oil, which is employed in preparing a cooling
liniment for headache [8]. An infusion of the plant is used to
treat cholera [9-10]. H. enneaspermus is also used for the
treatment of epilepsy, asthma, cough, cold, fever, stomach
problems and skin diseases [11]. H. enneaspermus, curd from
buffalo and cumin are mixed and given to animals to treat
diarrhea in veterinary medicine [12]. A paste of whole plant is
applied topically to treat cough [11]. The paste of two to three
entire plants of H. enneaspermus, with seven black peppers is
given once daily for the treatment of syphilis and dysentery
[13]. The juice of the whole plant is mixed with cow’s milk
and taken orally for a period of four to five months to get
relief from diabetes, and to improve the sexual vigor in the
male [14]. H. enneaspermus was also used by traditional heal-
ers to treat various types of complication like diabetes, ma-
laria, male sterility, urinary tract infections, and water reten-
tion [15-18]. The tender leaf stalks are used as demulcent;
whereas the roots are used as antigonorrhoeic, diuretic, for
bowel complaints and urinary problems. H. enneaspermus is
also used with food for pregnant and parturient women in
order to strengthen the child [4]. Formulations prepared from
the H. enneaspermus have been used externally for the treat-
ment of wounds and syphilis [19].
4 Pharmacognostical Features
4.1 Morphological characters
Roots of H. enneaspermus are spindle-shaped cylindrical,
rough and light yellow in colour. Stem is sparingly branched
with woody base and spreading erect branches. Leaves are
simple, alternate, sub-sessile, linear to lanceolate, 2.5 cm ×
0.7 cm, base nectariferous glands, apex acute. Flowers are 8
to 10 mm across, pink in colour, axillary, solitary and zygo-
morphic. Sepals are five in number, lanceolate, sub-equal,
obovate, upper ones oblong, laterals falcate, lower ones or-
bicular, clawed, and saccate at base. Petals are five, pink
coloured, unequal, upper ones oblong, laterals falcate long,
lower one larger, orbicular, clawed and saccate at base. Sta-
mens are five, connate, anterior filaments appendaged, pu-
berulous. Anthers are villous, while fruits are 5 mm across,
capsule, sub-globose with ribbed seeds. Powder is grayish
green, slightly odorous and has bland taste [20].
4.2 Microscopic characters
The epidermal cells of the leaf of H. enneaspermus is
composed of polygonal or elongated with straight, sinuous, or
arched, thick anticlinal walls. It also shows unicellular and
uniseriate-bicellular trichomes with vorrucoso margin. Sto-
mata are anisocytic, paracytic, anomocytic and transitional
between anisocytic and paracytic. The ontogony of the ani-
socytic and paracytic stomata is syndetocheilic or meso-
gonous, anomocytic is haploeheilie or perigonous, while that
of the transitional type is mesoperigenous [21]. The stem is
circular with uneven outline. The epidermis is uni-stratose
and thin, the cells are squarish with thin cuticle striations.
Hypodermis is collenchymatus 1−2 layered, cortex 5−6 lay-
ers of thin walled. Vascular cylinder consists of sclerids, nar-
row zone of phloem and dense cylinder of xylem. Pith is
wide and parenchymatous. Root has narrow uniform zone of
phellem and cortex. Cortex is composed of parenchyma,
secondary phloem. Fibres and prismatic crystals of calcium
oxalate are also present [22].
4.3 Powder characteristics
The powder is a pale green characteristic color with mu-
cilaginous taste, covering warty trichomes, paracytic stomata,
bundles of palisade cells, prism shape calcium oxalate crys-
tals, elongated fibres, tracheids and sclereid were found to be
present in the powder with oil droplets, and are the main
characteristic features of H. enneaspermus. The ash value of
the leaf and stem powder of H. enneaspermus showed higher
content of sulfated ash followed by total ash. The water solu-
ble extractive was high in both the stem and leaf powder of H.
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2013 年 5 月 第 11 卷 第 3 期 Chin J Nat Med May 2013 Vol. 11 No. 3 201

enneaspermus [22].
4.4 Comparative taxonomy
The genus Hybanthus Jacq. (Violaceae) comprises of
over 100 species of which most are mainly herbs, shrubs and
treelets distributed widely throughout the world [5]. H. en-
neaspermus was divided into two subgenera on the basis of
androecial characteristics and flower size viz. Hybanthus
(subgenera: Hybanthus) for the species commonly found in
the Northern and Central part of America and Hybanthus
(subgenera: Ionidium Vent.) which are of South American
origin [23]. The species H. enneaspermus itself has a number
of varieties, and some of the important ones are shown below
with their taxonomical differentiation [24]:
H. enneaspermus (var. enneaspermus): Annual or
perennial herb with a height of 30 cm tall, stems
simple or moderately branched in the lower half.
Leaf-lamina, linear to linear-lanceolate or lin-
ear-elliptic, 10−76 mm long, 1−10 mm wide. Flow-
ers are pink, magenta, reddish-violet, white, or bi-
coloured.
H. enneaspermus (var. densifolius): Leaves densely
congested, especially along the lateral shoots, often
more than 3 cm long.
H. enneaspermus (var. diversifolius): Perennial herb
up to 40 cm tall, stems erect to decumbent and
branched. Leaf-lamina linear-elliptic, 5−65 mm ×
2−12 mm, Pedicels 8−11 mm long. Lower petal
10−15 mm long.
H. enneaspermus (var. latifolius): Subshrub is up to 3
m tall, stems moderately branched. Leaf-lamina el-
liptic-lanceolate, 20−48 mm × 6−14 mm, acuminate.
Flowers are mauve or purplish. Pedicels 7−14 mm
long.
H. enneaspermus (var. nyassensis): Perennial herb,
30 cm tall, stems erect, branched from the base.
Leaf-lamina oblanceolate, 16−44 mm × 5−8 mm,
with prominent reticulate venation above. Flowers
are pink, purplish-lilac or violet. Pedicels 11−18 mm
long. Lower petal 12−20 mm long.
H. enneaspermus (var. pseudocaffer): Perennial herb
growing up to 60 cm height, stems erect or decum-
bent. Leaf-lamina narrow lanceolate-elliptic, acute,
26−85 mm × 5−13 mm, Flowers are reddish-purple.
Pedicels 20−26 mm long. Lower petal 12−15 mm
long.
H. enneaspermus (var. tsavoensis): Perennial herb up
to 24 cm in height; stems branched from the base,
otherwise simple, ascending., Leaf-lamina broadly
elliptic to elliptic-obovate appearing to be in fasci-
cles, 20−52 × 9−16 mm, Flowers are orange-red.
Pedicels 11−21 mm long. Lower petal 15−17 mm
long.
5 Phytochemistry
Preliminary phytochemical screening of H. enneasper-
mus showed the presence of steroids, triterpenes, sugars,
alkaloids, phenols, flavones, catechins, tannins, an-
thraquinones, and amino acids. Various phytoconstituents viz.
dipeptide alkaloids, aurantiamide acetate, isoarborinol, and
β-sitosterol, have been isolated from different parts of H.
enneaspermus [1, 22, 25-27]. H. enneaspermus contains a sig-
nificant amount of Fe and Mn content [28]. Various types of
amino acids, sugars, and flavonoids were found to be present
in H. enneaspermus which have been confirmed by paper
chromatography [29]. Five alkaloids (heteroxanthine, theo-
phylline, theobromine, caffeine, and tetramethyl trihydroxy-
purine), six steroids (agnosterol, lanosterol, dihydrositosterol,
cholestanol, cholesterol and dihydroagnosterol), three fla-
vonols (catechin, galangin and kaempferol), and six terpenoids
(gallic acid, thujyl alcohol, menthol, terpinol, sesquiterpene
and daucul) were isolated and identified from H. enneaspermus
by gas chromatography [27]. Phytoconstituents present in H.
enneaspermus are presented in Fig. 1.
6 Pharmacological Activity
The popularity of H. enneaspermus was due to the ideo-
logical belief in the treatment of multiple diseases. This plant
has occupied a special place for its wide range of medicinal
value in different systems of medicine. The plant has been
reported to have antimicrobial, antidiabetic, antiplasmodial, an-
timicrobial, anticonvulsant, nephroprotective, hepatoprotective,
antiinflamatory, aphrodisiac, aldose recductase inhibitory,
and free radical scavenging activities [30].
6.1 Anticonvulsant activity
Anticonvulsant activity of aqueous and ethanolic extract
of whole plant of H. enneaspermus at 200 and 400 mg·kg−1,
p.o. dose level using maximal electric shock and strych-
nine-induced convulsion models were investigated. Pheno-
barbitone sodium (30 mg·kg−1, p.o.) was used as a standard in
the study. The aqueous extract was found to have significant
anticonvulsant activity, whereas the ethanolic extract did not
show significant effect [31].
6.2 Antimicrobial activity
Petroleum ether, benzene, chloroform, and methanol ex-
tract of H. enneaspermus were tested for their antimicrobial
activity against two Gram positive bacteria, viz. Staphylo-
coccus aureus and Bacillus subtilis, and five Gram negative
bacteria, viz. Escherichia coli, Enterobacter aerogens, Pseu-
domonas aeruginosa, Salmonella typhi, and Citrobacter
freundii. Further, two active fractions were isolated from the
benzene extract of H. enneaspermus. From the result it was
found that the plant had significant antimicrobial activity,
which justifies its traditional use in urinary infections, wound
and burn infections, pneumonia, food poisoning, and derma-
titis. The difference in the antibacterial effect was mainly due
to differences in permeability barriers [32]. Furthermore, the
aqueous, ethanolic, petroleum ether, and chloroform extracts
of whole plant of H. enneaspermus were tested against Can-
dida albicans, Candida tropicalis and Candida krusei
through disc diffusion and micro broth dilution assays. From
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202 Chin J Nat Med May 2013 Vol. 11 No. 3 2013 年 5 月 第 11 卷 第 3 期

the obtained data, it was found that the ethanolic extract of H.
enneaspermus exhibited a significant and broader spectrum
of inhibition compare to the petroleum ether extract, which
showed moderate effect. However, the chloroform and aque-
ous extracts showed less significant activity compared to the
other fractions [33]. The aqueous, ethanolic, petroleum ether,
and chloroform extracts were tested against Escherichia coli,
Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus
mirabilis, Enterococcus faecalis, and Staphylococcus aureus
by the disc diffusion assay method. Among all of the frac-
tions, the ethanolic extract of H. enneaspermus was found to
have significant and broader spectrum of inhibition compared
to the aqueous extract. However, the chloroform and petro-
leum ether extracts were found to have less significant activ-
ity compared to the other fractions [34].


Fig. 1 Phytochemical constituents present in Hybanthus enneaspermus

6.3 Aphrodisiac activity
Aphrodisiac activity of the aqueous and alcoholic ex-
tracts of plant of H. enneaspermus at 300 mg·kg−1, p.o. dose
level in sexually inactive male rats, both in a single dose
regimen and in a chronic regimen for 28 days, were investi-
gated. Following a single dose administration, the aqueous
extract produced a decrease in the mounting and intromission
latency, with an increase in the ejaculatory and intromission
frequency. In the chronic model, both the alcohol and aque-
ous extracts increased the number of mounts, ejaculations
and intromissions with a decrease in the mounting and intro-
mission latency. Treatment with the aqueous extract also
elevated the testosterone levels in sexually inactive male rats
[35].
6.4 Anti-arthritic activity
The effects of the alcoholic and aqueous extracts of the
whole plant of H. enneaspermus on Freund’s adjuvant in-
duced arthritis was evaluated after 30 days of treatment at
500 mg·kg−1, p.o. in rats. Both of the extracts significantly
decreased the paw thickness. Though in the acute phase of
inflammation both of the extracts showed the same effect, in
the chronic phase the alcoholic extract was found to bemore
potent than the aqueous extract. From all of the obtained data
it was concluded that the aqueous and alcoholic extract of H.
enneaspermus possessed significant antiarthritic potential [10].
6.5 Nephroprotective activity
Effects of the alcoholic and aqueous extracts of whole
plant of H. enneaspermus were investigated for nephropro-
tective activity in a cisplatin-induced renal injury model in
rats at 250 and 500 mg·kg−1, p.o. dose level. The extracts
showed dose-dependent reduction in the elevated blood urea
and serum creatinine level. They increased glutathione (GSH),
glutathione S-transferase (GST), and super oxide dismutase
(SOD) levels and inhibited lipid peroxidation in the kidney
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homogenate [36].
6.6 Hypoglycemic activity
The hypoglycemic effects of the aqueous and methanolic
extracts of the leaf of H. enneaspermus at 80 and 180
mg·kg−1, p.o. in Wister rats for 12 h was evaluated. The
aqueous extract of H. enneaspermus gave a similar hypogly-
cemic result as glibenclamide. The potential hypoglycemic
activity of the aqueous leaf extracts of H. enneaspermus
could be beneficial in the treatment of diabetes [37]. Oral glu-
cose tolerance test and the normoglycemic effect of an alco-
holic extract of whole plant of H. enneaspermus were evalu-
ated at a dose of 125, 250, and 500 mg·kg−1, p.o. while the
hypoglycemic activity and effect on body weight were tested
at 250 and 500 mg·kg−1, p.o. for 21 days in streptozoto-
cin-induced diabetic rats. Further, the glucose uptake by
hemidiaphram was also evaluated. H. enneaspermus treat-
ment showed significant effect which resulted in increased
body weight and decreased blood glucose level. From the in
vitro study, it was found that alcoholic extract of whole plant
of H. enneaspermus also increased glucose uptake on isolated
rat hemi-diaphragm compared to control group [7].
6.7 Antioxidant activity
The alcoholic and aqueous extracts of whole plant of H.
enneaspermus were studied for their free radical scavenging
potential. The alcoholic and aqueous extracts of whole plant
of H. enneaspermus showed significant free radical scaveng-
ing effects on DPPH, ABTS, super oxide, nitric oxide, and
TBARS and ferric ion free radicals [36]. In vitro antioxidant
activity of a hydroalcoholic extract of the leaves of H. en-
neaspermus was investigated by using DPPH radical scav-
enging activity, hydrogen peroxide radical scavenging activ-
ity, reducing power activity, and nitric oxide radical scaveng-
ing activity. On the basis of the data obtained, it was con-
cluded that the leaf of H. enneaspermus has significant anti-
oxidant activity [38]. An alcoholic extract of the leaf of H.
enneaspermus was tested in terms of total antioxidant assay,
DPPH assay, reducing power, nitric oxide and hydrogen per-
oxide scavenging assays. From the obtained data it was found
that H. enneaspermus had significant antioxidant potential [7].
Petroleum ether, chloroform, ethyl acetate, and aqueous frac-
tions of H. enneaspermus were tested for antioxidant activity
in vitro in terms of total antioxidant assay, DPPH assay, re-
ducing power, nitric oxide, and hydrogen peroxide scaveng-
ing assays. Among the tested fractions, the ethyl acetate frac-
tion exhibited the higher total antioxidant capacity, a higher
percentage of DPPH radical scavenging activity, nitric oxide,
hydrogen peroxide, deoxyribose, and a higher reducing
power. This result revealed that the ethyl acetate fraction of H.
enneaspermus has strong antioxidant potential compared to
the other fractions [39]. Free radical scavenging activity of
aqueous and ethanolic extract of whole plant of H. en-
neaspermus was investigated in terms of the DPPH redical
scavenging assay. From the result, it was found that the
aqueous extract of H. enneaspermus had significant scav-
enging potential compared to the alcoholic extract [31].
6.8 Hepatoprotective activity
The hepatoprotective activity of the ethanolic extract of
leaves of H. enneaspermus at 200 mg·kg−1, p.o. in male al-
bino mice was investigated for 7 days. Enzymes such as CAT
(catalase), SOD (superoxide dismutase), GST (glutathione
S-transferase) and LPO (lipid peroxidase) were estimated in
the study. Histopathology of liver was studied to prove its
hepatoprotective effect. The H. enneaspermus extract restored
all of the altered parameters, which showed its hepatoprotec-
tive activity [40].
6.9 Anti-inflammatory activity
Anti-inflammatory activity of the alcoholic and aqueous
extracts of plant of H. enneaspermus against acute inflamma-
tion induced by carageenan, histamine, and 5-HT was inves-
tigated at the dose levels of 250 and 500 mg·kg−1, p.o. in rats.
Both extracts showed a dose-dependent decrease of paw
edema in the tested animals. The extracts also suppress the
inflammation induced by histamine and 5-HT. The alcoholic
extract showed a more significant effect compared to the
aqueous extract [41].
6.10 Aldose reductase inhibitory activity
Aldose reductase inhibitory activity of the petroleum
ether, chloroform, ethyl acetate, and aqueous fractions of
whole plant of H. enneaspermus were investigated in vitro
using rat lens aldose reductase enzyme. There was a signifi-
cant difference in the aldose reductase inhibitory activity of
each fraction. The ethyl acetate fraction showed maximum
inhibitory potential followed by the aqueous extract. The
least activity was in the petroleum ether fraction, whereas the
chloroform fraction showed moderate activity [42].
6.11 Antiplasmodial and larvicidal activity
The effect of different extracts of plant of H. en-
neaspermus was tested in vitro for antiplasmodial activity
towards Plasmodium falciparum K1 chloroquine resistant
and 3D7 chloroquine sensitive strains. From the obtained
data, it was found that the methylene chloride extract of H.
enneaspermus showed highly significant inhibition, with an
IC50 value 2.57 µmol·L−1 against the tested strains [16]. In an-
other study, the larvicidal activity of plant of H. enneaspermus
plant extract was tested against Culex quinquefasciatus and
was found to show significant larvicidal activity [43].
6.12 Toxicity studies
An acute toxicity study was carried out for the ethanolic
extract of whole plant of H. enneaspermus following OECD
425 guidelines. The H. enneaspermus extract was suspended
in 0.5% W/V CMC and was given at a dose of up to 5 000
mg·kg−1, p.o. body weight to overnight-fasted, healthy rats.
The animals were observed continuously for behavioral
changes, like abnormal locomotion, respiratory distress, and
uncoordinated muscle movements for 24 h. Alcoholic ex-
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204 Chin J Nat Med May 2013 Vol. 11 No. 3 2013 年 5 月 第 11 卷 第 3 期

tracts of H. enneaspermus produced no observable side ef-
fects up to 5 000 mg·kg−1 body weight in rats, even after 24 h
[7]. An oral acute toxicity test was performed for the alcoholic
and aqueous extract of whole plant of H. enneaspermus at
doses of 1 000, 2 000, 3 000, 4 000, and 5 000 mg·kg−1, p.o.
in albino mice weighing 20–25 g. After drug administration,
the animals were observed continuously for 2 h for gross
behavioral changes, and then intermittently once every 2 h.
and finally, at the end of 24 and 72 h, to note for any signs of
toxicity, including death. From the acute toxicity study, it was
found that the alcoholic and aqueous extracts of H. en-
neaspermus are safe up to 5 000 mg·kg−1[10]. Further, in an-
other acute oral toxicity study, the alcohol and aqueous ex-
tracts of plant of H. enneaspermus in increasing doses i.e. 30,
100, 300, 600, 1 000, and 3 000 mg·kg−1, p.o. were given to
rats. After administration of the drug, the rats were observed
continuously for 2 h for behavioral, neurological and auto-
nomic profiles. Further, at the end of 24 and 72 h, the rats
were observed for any lethality. In the acute toxicity study,
both the alcoholic and aqueous extracts of H. enneaspermus
produced no toxic effects or mortality in rats even at a dose
of 3 000 mg·kg−1[35]. From the obtained data it was concluded
that H. enneaspermus had very wide margin of safety.
7 Tissue Culture Techniques
A plant regeneration method for H. enneaspermus
through seed-derived callus was developed. Seeds were
demonstrated to have high induction frequency and a high
yield of light-yellow friable callus on MS (Murashige and
Skoog) medium containing 2.6 µmol·L−1 naphthaleneacetic
acids (NAA) and 2.2 µmol·L−1 benzyladenine (BA) within 4
weeks of incubation. After 1 year of subculture, yellow fri-
able and light-green compact callus was established. Shoot
differentiation was achieved from the light-green compact
callus. The addition of casein hydrolysate (500 mg·L−1) and
more potassium phosphate (1.86 mmol·L−1) to the culture
medium enhanced shoot differentiation. Rooting was
achieved on half-strength MS medium containing 4.8
µmol·L−1 indole-3-butyric acid (IBA). The regenerated plants
were morphologically uniform and exhibited a normal seed
set [44]. The highest number of shoot/explant was obtained
with initial pH values between 5.5 and 6.0. Similarly, shoot
differentiation occurred when light-green compact callus of H.
enneaspermus was transferred to MS medium supplemented
with 8.8 μmol·L−1 BA and 2.6 μmol·L−1 NAA; the highest
percentage of calli-forming shoots and the highest number of
shoots were achieved in this medium. Differentiated shoot
buds of H. enneaspermus elongated to 4−5 cm within 4
weeks. This response could be improved when glucose was
used as a carbon source for a larger flowered purslane, and
using higher concentrations of casein hydrolysate (500
mg·L−1) and potassium phosphate (1.86 mmol·L−1) for H.
enneaspermus [45]. It was also observed that an increase of the
NAA concentration augmented shoot length up to 8 cm of H.
enneaspermus in tissue culture [46].
8 Discussion
The present review discusses the different aspects related
to the traditional uses, morphology, microscopy, phytochem-
istry, pharmacology, and tissue culture technique of the me-
dicinal plant H. enneaspermus. Hence, this review will be
useful to researcher, having an interest in exploring and sci-
entifically validating the different traditional claims regarding
H. enneaspermus. Presently, there is an increasing interest
worldwide in herbal medicines due to the significant phar-
macological properties of the bioactive ingredients and their
ability to treat various diseases. Various types of drugs are
available in the market for the treatment of disease, which
came from the plant source after extensive research based on
the data available in the traditional system of the medicine [47].
Indigenous peoples in the world have unique and very rich
knowledge about the uses of plant materials for the treatment
of various types of disease. Due to their affordability and
effectiveness, traditional medicines are gaining in popularity
among all classes of people [48].
H. enneaspermus is one of the traditional ethnomedici-
nes used in different types of medicine in the world, includ-
ing in India for the treatment of various types of disorder.
Pharmacologically, H. enneaspermus was tested for various
types of pharmacological activities (Table 1). However, there
is still a lack of clinical data for its efficacy and safety profile.
H. enneaspermus contains various types of phytoconstituents
belonging to severalchemical classes. Few studies have been
carried out on the biological activity and medicinal applica-
tions of H. enneaspermus and its phytoconstituents. Scientific
investigation for its claimed pharmacological activities is
needed to make the plant more useful for the treatment of
different types of diseases. Even though different types of
preparation and extracts from various parts of H. enneasper-
mus have already been used and proven effective in the
treatment of various types of disease in different systems of
medicine studies such as the investigation of its bioactivity,
mechanism of action, pharmacotherapeutics, toxicity, stan-
dardization, and clinical trials should be undertaken from a
safety point of view before consumption. The world scenario
is changing towards the use of plant products for the treat-
ment of various types of disease in traditional, as well as in
modern, medicine. Scientific research should be undertaken
for H. enneaspermus for the treatment of various diseases.
There has been an increasing research on H. enneaspermus,
such as exploring the chemistry and pharmacological activity
of different parts of H. enneaspermus for the last few years,
still there is lot of unexplored areas of research associated
D. K. Patel, et al. /Chinese Journal of Natural Medicines 2013, 11(3): 199−206
2013 年 5 月 第 11 卷 第 3 期 Chin J Nat Med May 2013 Vol. 11 No. 3 205

with H. enneaspermus and its products for their better eco- nomic and therapeutic utilization.

Table 1 Pharmacological activities of Hybanthus enneaspermus
S. No. Type of extract Tested species Dose/(mg·kg−1), p.o. Activity reported References
1 Ethanolic extract of whole plant Rats 200 and 400 Anticonvulsant activity 31
2 Aqueous and alcoholic extract of plant Rats 300 Aphrodisiac activity 34
3 Alcoholic and aqueous extracts of plant Rats 250 and 500 Nephroprotective activity 35
4
Aqueous, methanolic and
alcoholic extract of whole
plant
Rats 80, 125, 180, 250 and 500 Hypoglycemic activity 7, 36
5 Alcoholic and aqueous extracts of whole plant Rats 500 Anti arthritic activity 10
6 Ethanolic extract of leaves Mice 200 Hepatoprotective activity 40
7 Alcoholic and aqueous extracts of plant Rats 250, 500 Antiinflamatory activity 41
8 Alcoholic extracts of whole plant Rats –
Aldose reductase inhibitory
activity 42
9
Petroleum ether, benzene,
chloroform, methanol, aque-
ous and ethanolic extract of
whole plant
Gram positive
and Gram nega-
tive bacteria
– Antimicrobial activity 31, 32, 33
10
Alcoholic, hydro alcoholic,
aqueous and ethanolic ex-
tracts, petroleum ether, chlo-
roform, ethyl acetate and
aqueous fraction of plant
– – Free radical scavenging activity 7, 30, 35, 37, 38
11 Methylene chloride extract of plant – –
Antiplasmodial and lar-
vicidal activity 16, 43

In conclusion, the present review provides an extensive
summary of the knowledge regarding different aspects such
as pharmacognosy, phytochemistry, pharmacology and tissue
culture techniques of H. enneaspermus. The data available
will help in the exploration of H. enneaspermus for its utili-
zation in human health care systems.
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