Fr e e R a d . A n t i ox .

 Basic Research Article

Comparative free radical scavenging potentials of different parts

of Solanum torvum

Thenmozhi A,1 Rao US Mahadeva2*

Department of Biochemistry, SRM Arts and Science College, Chennai– 603203, India
1

2
Faculty of Medicine and Health Science, Universititi Sultan ZainalAbidin, 20400 Kuala Terengganu, Terengganu, Malaysia

Submission Date: 31-10-2011; Revised Date: 2-12-2011; Accepted Date: 31-1-2012

ABSTRACT

Aim: An attempt has been made to evaluate phytochemicals and free radical scavenging efficacy of different parts of
Solanum torvum. Methods: The herbal powder obtained from various plant parts i.e.dry leaves, dry fruits and fresh
fruits were extracted with various solvents. The extracts were analysed for phytochemicals and antioxidants, carotenoids,
ascorbic acid, tocopherol, total phenol, proteins, reducing sugars and sterols. Free radical scavenging capacity was analysed
in terms of superoxide radial scavenging assay and reducing power assay. Result: Phytochemical characterization of the
different extracts revealed the presence of the phytochemicals-alkaloids, phenols, flavonoids, sterol, saponin glycosides,
reducing sugars, proteins, cardio active aglycones and cardinolides. Discussion: Excellent superoxide radical scavenging
ability found in almost all extracts of S. torvum. In the present study superoxide radical reduces nitroblue tetrazolium
(NBT) to a blue colored formazan that is measured at 560 nm. Antioxidant activity has been reported to be concomitant
with development of reducing power. This shows that extracts might contain reductones like ascorbic acid, reducing
sugar, thiol group containing protein which could react with free radicals to stabilize and terminate radical chain reaction.
Conclusion: These findings suggest that the promising phytonutrients of the plant could be exploited against oxidative
stress, cancer, ageing, Ischemic heart disease in dissolving thrombus, microbial infections and hormone replacement
therapy (HRT) justifying their use in traditional medicine as nutraceuticals.

Keywords: Solanum torvum, saponins, phytosterols, poly phenols, antimutagenic, cardinolides.

INTRODUCTION evaluation plays an im­portant role in the standardization

of crude herbal drugs.[3] Limited data from animal studies
The use of medicinal plants as source of the remedies for suggests that very high intake of phytosterols, particularly
the treatment of many diseases dates back to prehistory sitosterol, may inhibit the growth of breast[4] and prostate
and people of all continents have this old tradition.[1] In cancer.[5,6] Indole alkaloids exhibit numerous biological
developing countries where medicines are quite expen- activities such as anti-tumor, anti-microbial, anti-hyper-
sive, it is obvious that these medicinal plants will find their tensive and central nervous system stimulant.[7] A large
way in the arsenal of antimicrobial drugs.[2] Phytochemical body of scientific evidence associating dietary phyto-
chemicals with health and well-being of population has
stimulated tremendous activities to develop and commer-
*Corresponding address:
Rao US Mahadeva, Associate Professor, cialize products variously known as nutraceuticals, phy-
Faculty of Medicine and Health Science, Universititi Sultan toceuticals, dietary supplements, functional foods, etc.[8]
ZainalAbidin, 20400 Kuala Terengganu, Terengganu, Malaysia
Phone: 00 60 09 6275680 (Direct) During 1980s and 1990s, numerous laboratories began
Hand phone: 00 60 11 16547654 studying phytochemicals to “mine” plants for bioactive
Fax: 00 60 09 622 0021
E-mail: [email protected]
substances that might be used as medicines (nutraceuti-
cals) or for other chemical applications. Many compounds
DOI: 10.5530/ax.2012.2.5 are showing great promise as disease fighters in the body,

24  Free Radicals and Antioxidants Vol.2 / Issue 2 / Apr–Jun, 2012

 Thenmozhi A, et al.: Comparative free radical scavenging potentials of different parts of Solanum torvum

boosting production or activities of enzymes, which then MATERIALS AND METHODS

act by blocking carcinogens, suppressing malignant cells,
or interfering with the processes that can cause heart dis- Plant materials
ease and stroke.[9]
The leaves and fruits of Solanum torvum were collected
In the last few decades, several epidemiological studies have in Chennai, Tamil Nadu, India. The identification and
shown that a dietary intake of foods rich in natural antioxi- nomenclature of the plant was based on The Flora of
dants correlates with reduced risk of coronary heart disease Presidency of Madras[17] and The Flora of Tamil Nadu
particularly; a negative association between consumption of Carnatic.[18] They were later verified at Botanical Survey
polyphenol-rich foods and cardiovascular diseases has been of India, Southern Circle, Coimbatore, India. All the pre-
demonstrated. This association has been partially explained served specimens were deposited at the Herbarium of
on the basis of the fact that that polyphenols interrupt lipid Entomology Research Institute, Loyola College, Chennai.
peroxidation induced by reactive oxygen species (ROS).[10, 11]
Chemicals
Different stimuli, leading to an increase of ROS genera-
tion inside the cell, activate the phosphorylation of IkB All the solvents and chemicals were purchased from
inhibitory protein and the subsequent proteolysis. Thio- Sigma Laboratories, Bangalore, India.
redoxin may reduce activated NF-kB proteins facilitat-
Preparation of extracts for phytochemical analysis
ing nuclear translocation. Once released from IkB, the
NF-kB complex translocates into the nucleus and the The plant material (leaves and fruits) were air dried in the
binding to DNA domain in the promoters and enhancers laboratory at room temperature. It was then powdered
of genes such as TNF-a, IL-1, proliferation and chemo- and was extracted with hot water by boiling for 30 minutes
tactic factors, adhesion molecule. Some of these genes, to get the aqueous extract. The extract obtained was con-
in turn, may further induce NF-kB activation, leading to centrated and dried under controlled temperature (60oC).
a vicious circle if the regulatory cellular system escapes The dried powder was successively extracted with other
from control. Polyphenolic compounds as natural phy- solvents and kept in an orbital shaker for overnight. Fresh
tochemical antioxidants that possess anti-inflammatory fruit was homogenized with solvent and then extracted.
properties by down regulating NF-kB.[12-14] The obtained extract was filtered with Whatman No. 42
filter paper (125 mm) and the filtrate was collected and
A dramatic shift has taken place in consumer attitudes
used for experimental analysis.
toward food. An increasingly active population, the pur-
suit of healthier lifestyles and the desire to live longer has Phytochemical analysis
given rise to a new category of food products referred
to as “nutraceuticals.” Incorporated in the nutraceuticals These studies were performed according to the standard
category are functional foods. Functional foods can be methods[19-21] for alkaloids, glycosides-cardio active agly-
described as conventional foods containing ingredients cones, saponin glycosides, flavonoids, phenols, steroids,
that provide additional health or nutritional benefits proteins and reducing sugars.
beyond basic levels, leading to possible risk reduction of
contracting chronic disease.[15] Tests for free radical scavenging assay

Solanum torvum belongs to Solanaceae family (nightshade The antioxidant activity was evaluated using ethanol,
family), a spiny herb or shrub 3-4 m tall found throughout chloroform and aqueous extracts for non enzymatic anti-
the tropical parts of India and in the Andaman. Leaves oxidants, vitamin-C, vitamin-A, vitamin-E, phenol, pro-
are ovate, sinate or bilobed, lobes shallow, rarely deep; tein with -SH group, reducing carbohydrate and sterol.
flowers white, in dense lateral racemes, berries globere, Reducing power assay and super oxide radical scavenging
smooth, yellow or orange-red, seeds smooth. Whole plant assay was also performed with the above extracts.
is traditionally used as a digestive, diuretic and sedative.
Fruits are used for liver and spleen enlargement. Decoc- Estimation of vitamin-A was done by the method of
tion is used as antitussive. Leaves are used as haemostatic. Carr & Price.[22] Estimation of vitamin-C was done by
Root-paste is used in rhagades.[16] With this background, the method of Omaye.[23] Estimation of vitamin-E was
the present study was designed to screen for the presence done by Rosenberg method.[24] The reducing power of
of phytochemicals and free radical scavengers. the extract was determined according to the method of

Free Radicals and Antioxidants Vol.2 / Issue 2 / Apr–Jun, 2012 25

 Thenmozhi A, et al.: Comparative free radical scavenging potentials of different parts of Solanum torvum

Oyaizu.[25] The superoxide radical scavenging ability was glycosides, flavonoids, phenols, sterol, proteins, and car-
assessed by the method of Nishimik.[26] Total phenol bohydrates. Table 2 reveals that vitamin–(β.Carotene)
was determined using Lowry’s method.[27] Sterol was esti- content is highest in the leaf, high in dry fruit and less in
mated by Liebermann Burchard method.[28] fresh fruit. Chloroform extract has very high vitamin-A
than aqueous and ethanolic extract. Vitamin–A as a pow-
Statistical analysis erful, free radical scavenger (singlet oxygen) and chain
breaking antioxidant. The function of vitamin-A as a rad-
Experimental results are expressed as means ± SD. All
ical scavenging antioxidant can protect the cells from oxi-
measurements were replicated ten times. The data was
dative damage. Several clinical trials showed regression in
analyzed by an analysis of variance i.e. one way ANOVA
precancerous lesions of the cervix and the lung as well as
and student ‘t’ test using Graph Pad Quick Calcs.
the oral cavity with the administration of β-carotene.[29,30]
In this study it was found that vitamin-C (ascorbic acid)
RESULTS AND DISCUSSION content is in the following order.

The results of the phytochemical analysis were reported Leaf > Fresh Fruit > Dry Fruit; Aqueous Extract > Ethanolic
in Table 1. Extract > Chloroform Extract.

Table 1 shows the qualitative analysis results of the Vitamin-C is an excellent hydrophilic antioxidant; it
­phytochemicals of aqueous and organic extracts of Sola- readily scavenges ROS and peroxyl radical and also acts
num torvum which includes the presence of alkaloids, indole as a co-antioxidant by regenerating the vitamin- A, E
alkaloids, lepac alkaloids, cardio active aglycons, s­aponin and GSH from radicals.[31, 32] Table 2 indicates the high

Table 1  Qualitative analysis of the phytochemicals of aqueous and organic extracts of Solanum torvum
Phytochemicals analyzed Solanum torvum
Aqueous Ethanol Methanol Chloroform
Alkaloids +++ ++ ++ =/=
Indole alkaloids +++ +++ +++ =/=
Lepac alkaloids ++ + ++ =/=
Cardio active aglycons ++ ++ ++ =/=
saponin glycosides ++++ +++ +++ =/=
Flavonoids ++ + + =/=
Phenols ++++ +++ +++ =/=
Sterol +++ ++ ++ +++
Proteins +++ =/= =/= =/=
Carbohydrates +++ =/= =/= =/=
+ Presence of phytochemicals
++ Definite presence
+++ Definite heavy presence
++++ Definite heaviest presence
=/= Not done

Table 2  Level of different antioxidants in aqueous, ethanol and chloroform extracts of Solanum torvum per 100g
Parts of the Leaf Fresh Dry fruit Leaf Fresh Dry fruit Leaf Fresh Dry Fruit
plant fruit fruit fruit
Vitamin A 20.3 ± 6.12 ± 10.62 ± 25.51 ± 11.33 ± 12.76 ± 65.99 ± 14.62 ± 18.99 ±
(mg) 0.1491 0.1874 0.1874 0.2514 0.2983 0.2830 0.7578 0.1874 0.7578
Vitamin C 140.9 ± 130.8 ± 100.8 ± 180.9 ± 180.8 ± 130.8 ± 130.9 ± 95.8 ± 25.8 ±
(mg) 1.197 1.398 1.398 1.197 1.398 1.398 1.197 1.398 1.398
Vitamin E 20.84 ± 15.91 ± 10.77 ± 23.83 ± 13.47 ± 6.90 ± 26.91 ± 18.77 ± 11.33 ±
(mg) 0.1293 0.09816 0.1679 0.1366 0.3438 0.1035 0.09816 0.167 0.4342
Polyphenol 301.3 ± 151.3 ± 777.7 ± 651.3 ± 301.3 ± 101.7 ± 76.3 ± 52.8 ± 601.7 ±
(mg) 2.908 2.908 2.541 2.908 2.908 2.541 2.908 2.781 2.541
Protein 4.04 ± 3.54 ± 3.04 ± 0.64 ± 0.054 ± 0.44 ± 3.04 ± 2.54 ± 2.04 ±
(g) 0.1174 0.1174 0.1174 0.1174 0.1174 0.1174 0.1174 0.1174 0.1174
Carbohydrate 2.092 ± 2.204 ± 1.7 ± 2.004 ± 2.388 ± 1.8 ± 1.132 ± 1.18 ± 1.096 ±
(g) 0.0444 0.0672 0.1155 0.3967 0.0737 0.1155 0.0989 0.1155 0.07043
Values are mean ± SD of 10 replications.

26 Free Radicals and Antioxidants Vol.2 / Issue 2 / Apr–Jun, 2012

 Thenmozhi A, et al.: Comparative free radical scavenging potentials of different parts of Solanum torvum

antioxidants. Regeneration of vitamin E requires ascorbic

acid, an aqueous phase antioxidant.

Table 2 reveals that a high content of phenolic ­compounds

was found in aqueous extract indicating that phenolic
compounds in S. torvum were mainly soluble in water.
Phenolic compounds are likely to contribute to the radi-
cal scavenging activity of extract. Polyphenols are a large
class of compounds, occur naturally in food plants. The
flavonoids are the largest and best studied group of these.
Polyphenols are currently sold as dietary supplements
and/or herbal remedies. They have antioxidant, antimuta-
genic, anti-estrogenic, anti-carcinogenic and anti-inflam-
matory effects that might potentially be beneficial in
Figure 1.  level of sterol in different extracts of S.torvum in mg. preventing disease. Epidemiologic findings revealed that
polyphenols have cardio protective effects which include
inhibition of platelet aggregation and vascular relaxation
through the production of nitric oxide which decrease
LDL oxidation and prevent atherosclerosis.[33, 34]

Table 2 shows excellent protein content in ethanolic extract

of S. torvum. It may have more hydrophobic amino acids
and lesser hydrophilic amino acids containing proteins. So
they are extracted more in ethanol than in aqueous extract.
Amino acids, peptides, such as carnosine and anserine, and
proteins are common food components. Amino acids were
found to be efficient antioxidants in model experiments.
Their application is advantageous in mixtures with other
inhibitors as they often act as synergists of phenolic antioxi-
Figure 2.  Superoxide Radical Scavenging of Solanum torvum in%. dants and as chelating agents. Amino acids convert hydro-
peroxides into imines, and sulphur containing amino acids
reduce hydroperoxides into the respective inactive hydrox-
ylic derivatives. Methionine and selenomethionine were
found to be more active than a-tocopherol in olive oil.[35]

Table 2 shows that S. torvum has high carbohydrates in

aqueous extract indicating that carbohydrates in S. to­r vum
were soluble in water. These are likely to contribute to
the radical scavenging activity and reducing power of the
extract. Simple carbohydrates have antioxidant proper-
ties. Hydroxyl radicals generated by a Fenton reaction
induce damage on simple carbohydrates with a conse-
quent free radical scavenging activity. Carbohydrate activi-
ties were measured by different methods as spin-trapping
of hydroxyl radical and electron paramagnetic resonance
Figure 3.  Reducing power of different extract of S.torvun at detection and 1, 1-diphenyl-2-picrylhydrazyl quenching.
700 nm. Carbohydrate damage was evaluated in a Fenton system
by measuring the reactive substances to thiobarbituric
vitamin-E content in chloroform extract. Vitamin-E is acid, by their decreased detection with an HPLC test, and
a fat soluble vitamin which can be extracted effectively by a gas chromatographic determination of formic acid
by chloroform. S. torvum has very high vitamin-C along from sugar oxidation. Different intensities of damage and
with vitamin-E content. Vitamin C and E are synergistic scavenging were found according to molecular structure,

Free Radicals and Antioxidants Vol.2 / Issue 2 / Apr–Jun, 2012 27

 Thenmozhi A, et al.: Comparative free radical scavenging potentials of different parts of Solanum torvum

and some hyphotheses on the carbohydrate action against Isoflavones have a role similar to hormones, and act as
free radicals were attempted. The assayed disaccharides phytoestrogens. They benefit humans in four ways as
were shown to be more active toward and less damaged cancer enzyme inhibitor, as antioxidants and as immune
by hydroxyl radical than monosaccharides.[36] system enhancers or stimulants. Phytoestrogens compete
with estrogen for binding to estrogen receptors. Their
Table 2 and Figure 3 indicate that S. torvum leaf, fresh fruit use could have beneficial effects on preventing osteo-
and dry fruit have excellent sterol content both in organic porosis and sex hormone-mediated malignancy such as
and aqueous extracts. This may be due the presence of breast and prostate cancer. Clinical trials identified the
sterol which is extractable with organic and aqueous sol- potential efficacy of isoflavones in the prevention of
vents. S. torvum was found to have steroidal saponins and coronary heart disease, osteoporosis, breast and prostate
steroidal alkaloid which contributes total sterol content. cancer.[40, 41]
S. torvum has β-sitosterol- an important group of anti-
oxidant. Plant sterols inhibit the intestinal absorption of Figure 2 shows the excellent superoxide radical scaveng-
cholesterol. Functional food ingredient derived from phy- ing ability found in almost all extracts of S. torvum. Super
tosterols has been clinically proven to have significantly oxide anions are one of the potent reactive oxygen species
lower Low Density Lipoprotein (LDL) or “bad” choles- (ROS) which are produced from molecular oxygen due
terol when consumed in different foods. So they have to oxidative enzymes[42] of body as well as via non enzy-
a hypocholesterolemic action. They also inhibit endog- matic reaction such as autoxidation by catecholamine.[43]
enous synthesis of cholesterol by inhibiting and repress- In the present study superoxide radical reduces NBT to
ing the rate limiting enzyme Hydroxy methyl glutaryl coA a blue colored formazan that is measured at 560 nm[44]
(HMG-CoA) reductase in cholesterol synthesis. Steroidal Figure 3 shows the excellent reducing power of Solanum
glycosides in plant foods have estrogenic antiestrogenic torvum. In the reducing power (Fe3+- Fe2+ transformation
actions and are known as phytoestrogen. These have anti- ability) assay, the presence of antioxidants in the extracts
bacterial and antifungal actions. They also produce typi- would result in the reducing of Fe 3+ to Fe2+ by donat-
cal estrogen responses with a biological activity 1/500 to ing an electron. Amount of Fe2+ complex can be then be
1/1000 of estradial. Studies proved that phytoestrogens monitored by measuring the formation of Perl's Prussian
lower the incidence of osteoporosis, breast and uterine blue at 700 nm [45]. Antioxidant activity has been reported
cancer.[37] to be concomitant with development of reducing power.
This shows that extracts might contain reductones, like
Saponins are natural surfactants or detergents. It can ascorbic acid, reducing sugar, thiol group containing pro-
be used as foaming agents for beverages. Saponins tein which could react with free radicals to stabilize and
have astypic activity which can be utilized in dissolv- terminate radical chain reaction.
ing thrombus. Recent studies have suggested that the
low serum cholesterol levels of Masai tribes in East CONCLUSION
Africa who consume a diet very high in animal prod-
ucts, cholesterol and saturated fat with saponin rich- These findings suggest that the promising antioxidant,
herbs. Saponins act by binding with bile acids and ­cardio active (aglycones), astypic, antimitotic, anti micro-
cholesterol. So it cleans or purges fatty compounds bial and antitumour properties of the plant could be
from the body, lowering the blood cholesterol level. exploited in herbal preparations against oxidative stress,
Digitalis is a saponin used as heart tonic to strengthen cancer, ageing, Ischemic heart disease in dissolving throm-
contractions of the heart muscle. Saponins are active bus, microbial infections and hormone replacement ther-
antifungal agents.[38] apy (HRT) justifying their use in traditional medicine. The
plant extracts have assumed an increased importance in
The alkaloid present in Solanum torvum is an indole alka- medicine and in health care industry and further work on
loid. The aromatic nitrogen heterocyclic is a potent the above suggested aspects may be given prominence.
cancer fighter/anticancer agent, blocking carcinogenic Phytochemicals occur naturally in plant foods which cre-
substances before they reach their cellular targets and ate an entirely new philosophy of “functional foods,”
eliminating DNA damage in cell nuclei. It may also turn ­eating not just to sustain minimal basic health but also
out to be an important chemical tool in fighting breast eating to prevent disease. In the future, we may tailor our
cancer because it inhibits estrogen induced growth of diets to include the foods that will best address our per-
cancer cells and converts the more dangerous forms of sonal health problems and risks as well as maintain opti-
estrogen to safer forms.[39] mal health.

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 Thenmozhi A, et al.: Comparative free radical scavenging potentials of different parts of Solanum torvum

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