Phytochemical and Antimicrobial Efficacy of Successive Series of Solanum
xanthocarpum
Sudhanshu1, Mittal Sandhya1, Rao
Nidhi1, Menghani Ekta2*
1Suresh Gyan Vihar University, Jaipur. 22.
India.
2Mahatma
Gandhi Institute of Applied Sciences, JECRC University, Jaipur-22. India.
ABSTRACT:
Antimicrobial
activity of the extracts of Solanum xanthocarpum against bacteria and fungus was evaluated.
Plant extracts of S. xanthocarpum were
prepared in distilled water and in organic solvents, petroleum ether, chloroform, benzene, ethyl
acetate and ethanol. Disc diffusion method technique was used to assess the
antimicrobial activity of various extracts against Shigella
flexneri, Staphylococcus aureus,
Salmonella typhi, Pseudomonas aeruginosa,
Klebsiella pneumonia, Proteus vulgaris,
Enterobacter aerogenes, Aspergillus niger , Candida albicans, Trichophyton rubrum. The diameter of zone of inhibition was taken as an indicator of
antimicrobial effect. Except aqueous extracts of S. xanthocarpum,
extracts prepared in organic solvents showed antimicrobial activity against the
test organisms. Methanol extract showed the
best activity against all test microorganisms than other solvent extracts.
The maximum efficacy of methanol extract A4 (200mg concentration) was against Staphylococcus aureus
(I.Z.-23mm) was screened. Thus, S. xanthocarpum could be considered as a potential source
of natural antimicrobials.
KEYWORDS: Solanum xanthocarpum, antimicrobial activity, organic solvents.
INTRODUCTION:
More than the last few
decades, a enormous attention has residential in
pointed for antimicrobial drugs from natural plant products. This notice
chiefly arises from the faith that drugs copied from plants are protected and
reliable compared with synthetic drugs that may contain adverse effects on host
as well their high cost. Natural antimicrobials came from a broad collection of
sources counting plants, animals and microorganisms (Gordon and David, 2001).
Researchers have so far discovered approximately over 10,000 biologically
active compounds of microbial origin (Shahidi et al., 2004). Recently, a lot of
bacterial pathogens are becoming resistant to existing antibiotics owing to
their indiscriminate use in the action of infectious diseases (Davis, 1994;
Service, 1995; Shears, 2000). So, there is a demand to find out new and
well-organized antimicrobials from other sources such as plants (Cordell, 2000;
Karaman et al.,
2003; Raghavendra et al., 2005).
Solanum xanthocarpum has thoughtful use in Ayurveda
and folklore medicine. It is a normally rising perennial herbaceous weed with
bright green leaves and zig-zag stem, mostly found in
the arid region. It is hypothetical that the plant has solasonine
in its diverse parts, which is accountable for its medicinal value (Oudhia, 2007). In the present study an attempt was made to
screen different extracts prepared from S. xanthocarpum
for its antimicrobial action against bacteria and fungi.
MATERIALS
AND METHODS:
Collection:
Plant sample Solanum xanthocarpum was collected from various tribes living in
tribal pockets of Mt. Abu, arid zone of Rajasthan, in the month of July, 2009. These
plants were used by these tribes in their daily lives to cure various ailments.
Identification:
These samples were
authenticated and submitted in Ethnomedicinal
Herbarium, Centre of Excellence funded by DST, MGiaS,
Jaipur (Rajasthan).
Sources of test organisms:
Bacteria-Pure culture of all
test organisms, namely Pseudomonas aeruginosa, Staphylococcus aureus,
Klebsiella pneumoniae,
Salmonella typhi, Shigella flexneri, Proteus vulgaris, Enterobactor aerogenes and
fungi Candida albicans,
Aspergillus niger, Trichophyton rubrum were obtained through the courtesy of Mahatma
Gandhi Institute of applied Sciences (MGiaS), Jaipur, which were maintained on Nutrient broth media. Culture
of test microbes: For the cultivation of bacteria, Nutrient Agar Medium (NAM)
was prepared by using 20 g Agar, 5 g Peptone, 3 g beef extract and 3 g NaCl in 1 L distilled water and sterilized at 15 lbs
pressure and 121°C temperature for 25-30 min. Agar test plates were prepared
pouring approximately 15 ml of NAM into the Petri dishes (10 mm) under aseptic
conditions. A saline solution was prepared (by mixing 0.8% NaCl)
in distilled water, followed by autoclaving and the bacterial\ cultures were
maintained on this medium by regular sub-culturing and incubation at 37°C for
24-48 h. To prepare the test plates, in bacteria, 10-15 ml of the respective
medium was poured into the Petri plates and used for screening. For assessing
the bactericidal efficacy, a fresh suspension of the test bacteria was prepared
in saline solution from a freshly grown Agar slant.
Preparation of test extracts:
Crushed powders of species
were successively soxhlet extracted. Later, each of
the homogenates was filtered and the residue was re-extracted twice for
complete exhaustion, the extracts were cooled individually. Each filtrate was
concentrated to dryness in vitro and re dissolved in respective solvents, were
stored at 4°C in a refrigerator, until screened for antibacterial activity.
Bactericidal assay:
For both, bactericidal in
vitro Disc diffusion method was adopted 2, because of
reproducibility and precision. The different test organisms were proceeded
separately using a sterile swab over previously sterilized culture medium
plates and the zone of inhibition were measured around sterilized dried discs
of Whattman No. 1 paper (6 mm in diameter), which
were containing three different concentration, its control (of the respective
solvent) and tetracycline as reference drugs (standard disk) separately. Such
treated discs were air-dried at room temperature to remove any residual
solvent, which might interfere with the determination, sterilized and
inoculated. These plates were initially placed at low temperature for 1 h so as
to allow the maximum diffusion of the compounds from the test disc into the
agar plate and later, incubated at 37°C for 24 h in case of bacteria, after
which the zones of inhibition could be easily observed. Five replicates of each
test extract were examined and the mean values were then referred.
The Inhibition Zone (IZ) in
each case were recorded and the Activity Index (AI) was calculated as compared
with those of their respective standard reference drugs (AI = Inhibition Zone
of test sample/Inhibition zone of standard).
Phytochemical Screening:
Phytochemical screening was
performed using standard procedure:
Test for reducing sugars
(Fehling’s Test):
The aqueous ethanol extract
(0.5gm in 5 ml of water) was added to boiling Fehling’s solution (A and B) in a
test tube. The solution was observed for a colour
reaction.
Test for terpenoides (Salkowski Test)-:
To 0.5 gm each of the
extract was added to 2ml of chloroform. Concentrated sulphuric
acid (3ml) was carefully added to form a layer. Reddish brown coloration of the
interface indicates the presence of terpenoides.
Test for flavonoides:
4ml of extract solution was
treated with 1.5ml of 50% methanol solution. The solution was warmed and metal
magnesium was added. To this solution, 5-6 drops of concentrated Hydrochloride
acid was added and red colour was observed for flavonoids and orange color for flavons.
Test for tannins:
About 0.5 g of the extract
was boiled in 10ml of water in a test tube and then filtered. A few drops of
0.1% ferric chloride was added and observed for brownish green or a blue-black
coloration.
Test for saponins:
To 0.5 g of extract was
added 5 ml of distilled water in a test tube. The solution was shaken
vigorously, and observed for a stable persistent froth. The frothing was mixed
with 3 drops of olive oil and shaken vigorously after which it was observed for
the formation of an emulsion.
Test for alkaloids:
Alkaloids solutions produce
white yellowish precipitate when a few drops of Mayer’s reagents are added.
Most alkaloids are precipitated from neutral or slightly acidic solution by
Mayer’s regent. The alcoholic extract was heated on a boiling water bath with
2% hydrochloric acid. After cooling, the mixture was filtered and treated with
a few drops of Mayer’s reagent. The sample was then observed for the turbidity
or yellow precipitation.
Table
1: Preliminary phytochemical screening of the plant Solanum xanthocarpum. [(+:
present) (- : absent).
|
PLANTS |
Reducing sugar |
Terpenoides |
Flavonoids |
Tannin |
Saponin |
|
Extracts |
|
||||
|
Pet.
ether |
+ve |
+ve |
-ve |
-ve |
+ve |
|
Benzene |
+ve |
+ve |
-ve |
+ve |
+ve |
|
Chloroform |
+ve |
+ve |
-ve |
+ve |
-ve |
|
Ethyl
acetate |
+ve |
-ve |
-ve |
+ve |
+ve |
|
Methanol |
+ve |
-ve |
-ve |
+ve |
-ve |
|
Distilled
water |
+ve |
+ve |
+ve |
+ve |
-ve |
Table No. 2: Anti
microbial activity of the petroleum ether, chloroform, benzene, ethyl
acetate and ethanol and distilled water extracts of Solanum xanthacarpum.
|
|
|
|
|
Sf |
Sa |
St |
Pv |
Kp |
Pa |
Ea |
An |
Ca |
Tr |
|
1 |
Pet. Ether |
A1 |
I.Z. |
0 |
6 |
6 |
0 |
7 |
7 |
0 |
12 |
0 |
6 |
|
|
|
|
A.I. |
0 |
0.30 |
0.20 |
0 |
0.17 |
0.43 |
0 |
0.50 |
0 |
0.21 |
|
|
|
A2 |
I.Z. |
7 |
7 |
7 |
8 |
7 |
7 |
9 |
0 |
7 |
7 |
|
|
|
|
A.I. |
0.20 |
0.35 |
0.23 |
0.18 |
0.17 |
0.43 |
0.21 |
0 |
0.18 |
0.25 |
|
|
|
A3 |
I.Z. |
7 |
6 |
7 |
8 |
7 |
7 |
10 |
21.3 |
7 |
9 |
|
|
|
|
A.I |
0.20 |
0.30 |
0.23 |
0.18 |
0.17 |
0.43 |
0.24 |
0.88 |
0.18 |
0.32 |
|
2 |
Benzene |
A1 |
I.Z. |
7 |
0 |
8 |
8 |
9 |
8 |
0 |
7 |
8 |
7 |
|
|
|
|
A.I. |
0.20 |
0 |
0.26 |
0.18 |
0.22 |
0.50 |
0 |
0.29 |
0.21 |
0.25 |
|
|
|
A2 |
I.Z. |
8 |
8 |
9 |
9 |
8 |
8 |
0 |
7 |
8 |
7 |
|
|
|
|
A.I. |
0.22 |
0.40 |
0.30 |
0.20 |
0.18 |
0.50 |
0 |
0.29 |
0.21 |
0.25 |
|
3 |
Chloroform |
A1 |
I.Z. |
12 |
9 |
11 |
8 |
8 |
0 |
0 |
0 |
12 |
9 |
|
|
|
|
A.I. |
0.34 |
0.45 |
0.36 |
0.18 |
0.20 |
0 |
0 |
0 |
0.32 |
0.32 |
|
|
|
A2 |
I.Z. |
15 |
12 |
19 |
19 |
18 |
9 |
12 |
0 |
19 |
10 |
|
|
|
|
A.I. |
0.62 |
0.60 |
0.63 |
0.43 |
0.40 |
0.56 |
0.29 |
0 |
0.51 |
0.35 |
|
4 |
Ethyl acetate |
A1 |
I.Z. |
0 |
8 |
9 |
9 |
9 |
8 |
0 |
8 |
10 |
8 |
|
|
|
|
A.I. |
0 |
0.40 |
0.30 |
0.20 |
0.22 |
0.50 |
0 |
0.33 |
0.27 |
0.28 |
|
|
|
A2 |
I.Z. |
9 |
0 |
10 |
9 |
0 |
9 |
8 |
7 |
10 |
8 |
|
|
|
|
A.I. |
0.25 |
0 |
0.33 |
0.20 |
0 |
0.56 |
0.19 |
0.29 |
0.27 |
0.28 |
|
|
|
A3 |
I.Z. |
8 |
13 |
9 |
9 |
8 |
8 |
9 |
8 |
9 |
9 |
|
|
|
|
A.I. |
0.22 |
0.65 |
0.30 |
0.20 |
0.20 |
0.50 |
0.21 |
0.33 |
0.24 |
0.32 |
|
5 |
Methanol |
A1 |
I.Z. |
11 |
14 |
9 |
10 |
11 |
11 |
9 |
11 |
14 |
14 |
|
|
|
|
A.I. |
0.31 |
0.70 |
0.30 |
0.22 |
0.27 |
0.68 |
0.21 |
0.45 |
0.37 |
0.50 |
|
|
|
A2 |
I.Z. |
11 |
17.3 |
13.3 |
10 |
9 |
14 |
8 |
8 |
9 |
11 |
|
|
|
|
A.I. |
0.31 |
0.86 |
0.44 |
0.22 |
0.22 |
0.87 |
0.19 |
0.33 |
0.24 |
0.39 |
|
|
|
A3 |
I.Z. |
13 |
20 |
11 |
14 |
15 |
14 |
8 |
0 |
19 |
17 |
|
|
|
|
A.I |
0.37 |
1 |
0.36 |
0.31 |
0.37 |
0.87 |
0.19 |
0 |
0.51 |
0.60 |
|
|
|
A4 |
I.Z. |
0 |
23 |
11 |
10 |
15 |
13 |
8 |
8 |
13 |
11 |
|
|
|
|
A.I. |
0 |
1.15 |
0.36 |
0.22 |
0.37 |
0.81 |
0.19 |
0.33 |
0.35 |
0.39 |
|
|
|
A5 |
I.Z. |
13 |
19 |
19 |
11 |
13 |
15 |
8 |
7 |
19 |
19 |
|
|
|
|
A.I. |
0.37 |
0.95 |
0.63 |
0.25 |
0.32 |
0.93 |
0.19 |
0.29 |
0.51 |
0.67 |
|
6 |
D. Water |
A1 |
I.Z. |
13 |
9 |
10 |
0 |
8 |
10 |
10 |
10 |
10 |
13 |
|
|
|
|
A.I. |
0.37 |
0.37 |
0.33 |
0 |
0.20 |
0.62 |
0.24 |
0.50 |
0.27 |
0.46 |
|
|
|
A2 |
I.Z. |
13 |
8 |
10 |
0 |
0 |
8 |
13 |
10 |
9 |
10 |
|
|
|
|
A.I. |
0.37 |
0.33 |
0.33 |
0 |
0 |
0.50 |
0.31 |
0.50 |
0.24 |
0.35 |
|
|
|
A3 |
I.Z. |
14 |
9 |
10 |
0 |
9 |
8 |
11 |
11 |
10 |
12 |
|
|
|
|
A.I |
0.40 |
0.37 |
0.33 |
0 |
0.22 |
0.50 |
0.26 |
0.55 |
0.27 |
0.42 |
|
|
|
A4 |
I.Z. |
15 |
10 |
11 |
8 |
9 |
8 |
12 |
10 |
11 |
12 |
|
|
|
|
A.I. |
0.42 |
0.41 |
0.36 |
0.18 |
0.22 |
0.50 |
0.29 |
0.50 |
0.29 |
0.42 |
|
|
|
A5 |
I.Z. |
15 |
10 |
13 |
0 |
7 |
8 |
11 |
9 |
11 |
12 |
|
|
|
|
A.I. |
0.42 |
0.41 |
0.43 |
0 |
0.17 |
0.50 |
0.26 |
0.45 |
0.29 |
0.42 |
RESULTS
AND DISCUSSION:
Phytochemical
screening:
Phytochemical analysis for
Solanum xanthocarpum
extract was performed and the phyto constituents
reported in the Table no.1.
The result of the phytochemical screening of Solanum xanthacarpum (Pasarkateli) is presented in Table
2. This reveals moderate concentration of reducing sugar, flavonoids,
saponins, terpenoids and
tannin in different extraction solvents, some of which chemical compounds have
been associated to antimicrobial activities and thus have curative properties
against selected bacteria and fungi. Standard
method were used for preliminary phytochemical
screening of the extract was performed to know the phyto-constituents
in the extract and it was found that petroleum extract contains reducing sugar,
terpenoids and saponins,
benzene extract contains reducing sugar, terpenoids,
tannin and saponins, chloroform extract contain
reducing sugar, terpenoids and tannin, ethyl acetate
extract contains reducing sugar, tannin and saponins,
methanol extract contains reducing
sugar and tannin and distilled water extract contains redusing sugar, terpenoids,
tannin and flavonoids.
Antimicrobial
Activity:
Antimicrobial activity of
extracts from Solanum xanthacarpum
was tested against Shigella flexneri, Staphylococcus aureus,
Salmonella typhi, Pseudomonas aeruginosa,
Klebsiella pneumonia, Proteus vulgaris,
Enterobacter aero genes, Aspergillums Niger, Candida albicans, Trichophyton rubrum by using the disc diffusion method. The plates
containing the micro organisms were then perforated and the disc were placed
with 50mg/20disc,100mg/20disc,150mg/20disc,200mg/20disc,250mg/20disc
concentrations. Microbial growth was determined by measuring the diameter of
zone of inhibition.
Sf-Shigella flexneri,
Sa- Staphylococcus aureus, St-Salmonella typhi,
Pv- Proteus vulgaris, Kp- Klebsiella pneumoniae,
Pa- Pseudomonas aeruginosa, Ee-Enterobacter aerogenes,
An- Aspergillus niger, Ca-Candida albicans,
Tr- Trichophyton rubrum ; 0 - no inhibition zone.
In the present investigation, the petroleum ether, chloroform,
benzene, ethyl acetate and ethanol and distilled water extracts exhibited
nearly similar considerable antimicrobial activity indicating the suitability
of these solvents for dissolving most of the bioactive compounds of the plants.
All the extracts highly affected the activity of bacteria and fungi. Methanol
extract showed the best activity against all test microorganisms than other
solvent extracts. The maximum efficacy of pet.ether extract A3 (150mg concentration) was against Aspergillus niger
(I.Z.-21.33mm), Enterobacter aerogenes
(I.Z.-10mm), Trichophyton rubrum (I.Z.-9mm)
The maximum efficacy of benzene extract A1 (50mg concentration) was against Klebsiella pneumoniae
(I.Z.-9mm), Salmonella typhi (I.Z.-8mm), whereas no activity against Enterobacter aerogenes has
been screened.
The
maximum efficacy of chloroform extract A2 (100mg concentration) was against Salmonella typhi
(I.Z.-19mm), Proteus vulgaris
(I.Z.-9mm), Candida albicans
(I.Z.-19mm), Klebsiella pneumoniae
(I.Z.-18mm), Shigella Flexneri
(I.Z.-15mm) whereas no activity against Aspergillus niger has been screened. The
maximum efficacy of ethyl acetate extract A3 (150mg concentration) was against Staphylococcus aureus
(I.Z.-13mm), Salmonella typhi (I.Z.-9mm), Proteus
vulgaris (I.Z.-9mm), Enterobacter aerogenes (I.Z.-9mm) and the maximum
efficacy of methanol extract A4 (200mg concentration) was against Staphylococcus aureus
(I.Z.-23mm), Klebsiella pneumoniae
(I.Z.-15mm), Pseudomonas aeruginosa (I.Z.-13mm)
and A3 (150mg concentration) was active against Staphylococcus aureus (I.Z.-20mm), Candida albicans
(I.Z.-19mm), Trichophyton rubrum
(I.Z.-17mm) and A5 (250mg concentration) was also active against Staphylococcus aureus
(I.Z.-19mm), Candida albicans
(I.Z.-19mm), Trichophyton rubrum
(I.Z.-19mm) and A2 (100mg concentration) was active against Staphylococcus aureus
(I.Z.-17.33mm), and the maximum
efficacy of distilled water extract A4 (200mg concentration) was against Shigella flexneri
(I.Z.-15mm), Enterobacter aerogenes
(I.Z.-12mm), Trichophyton rubrum
(I.Z.-12mm) and A5 (250mg concentration)
was active against Shigella flexneri
(I.Z.-15mm), Salmonella typhi (I.Z.-13mm)
have been screened ,even there is good activity against all the
microorganisms (bacteria and fungi).
Fig-1. Antibacterial activity of petroleum ether, benzene,
chloroform, ethyl acetate, ethanol and distilled water extract of Solanum xanthacarpum.
A1-50 mg/20 disc, A2-100 mg/20
disc, A3-150mg/20 disc, A4-200mg/20 disc, A5-250mg/20 disc. I.Z. - Inhibition zone, I.A. – Activity
index.
There are many
information on the antimicrobial action of S. xanthocarpum
extracts in the literature. likewise, Javanmardi et al. (2003) reported that the sum phenolic content of different accessions of Ocimum are in the range of 6.07 to 65.5 mg
GAE per dry weight. The total phenolics of alcoholic
extracts may purpose as physically powerful antimicrobial compound(s) against
the tested microorganisms.
Additional
work on separation of various plant metabolites is necessary to determine the
antimicrobial potential of S. xanthocarpum. It
is obvious from the in attendance study that the extracts of examined parts of S.
xanthocarpum were active against the tested
microorganisms. Based these result and the medicinal potential of this plant,
we put forward that additional phytochemical studies
be performed to decide the chief active principles accountable for the
antimicrobial result of this plant. The results hold up the use of this plant
in folklore medicine for action of infectious diseases.
ACKNOWLEDGEMENT:
Author
acknowledge with thanks the financial support from Department of Science and
Technology, Government of Rajasthan, in the form of Centre with Potentials for
Excellence in Biotechnology, sanction no F 7(17) (9) Wipro/Gaprio/2006/7358-46(31/10/2008).
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Received on 24.05.2012
Modified on 28.05.2012
Accepted
on 30.05.2012
©
A&V Publication all right reserved
Research Journal of Pharmacognosy
and Phytochemistry. 4(4): July- August 2012, 215-219