Int. J. Pharm. Sci. Rev. Res., 26(2), May – Jun 2014; Article No. 47, Pages: 273-281
ISSN 0976 – 044X
Research Article
Antimicrobial Activity and Phytochemical Analysis of Urginea Indica from
Bastar District of Chhattisgarh
Dhananjay Pandey*, Ashwini Kumar Gupta
Microbiology Research Laboratory, School of Studies in Life Sciences, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India.
*Corresponding author’s E-mail: pandey.dhananjay333@gmail.com
Accepted on: 12-04-2014; Finalized on: 31-05-2014.
ABSTRACT
Medicinal plants are the bio-resources for the discovery of novel bioactive compounds. The present investigation was carried out to
explore the antimicrobial activity and phytochemical analysis of Urginea indica (family: Liliaceae) commonly known as jungli piyaz.
The root, stem and leaf of Urginea indica were extracted successively with polar (aqueous, methanol), dipolar (acetone) and non
polar (chloroform) solvents, and their physical characteristics were evaluated which revealed presence of more Phytocompounds in
root followed by stem and leaf. The extracts were assessed for their antimicrobial activity against four gram positive and four gram
negative bacteria viz., Bacillus cerus, Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Proteus
vulgaris, Pseudomonas aeruginosa, Klebsiella pneumoneae and against two fungi viz., Aspergillus niger and Candida albicans. The
gram positive bacteria were more susceptible than gram negative bacteria. The inhibition of gram positive, gram negative bacteria
and fungi by the extracts indicates the presence of broad spectrum antimicrobial potentiality of the plant. The root extracts showed
inhibition against Bacillus cerus, Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Klebsiella
pneumoneae and against fungi viz., Aspergillus niger and Candida albicans. The highest activity index was recorded in root methanol
extracts in Bacillus cerus and acetone extracts in case of Pseudomonas aeruginosa respectively, whereas root acetone extract
exhibited pronounced activity against Aspergillus niger and Candida albicans. The phytochemical analysis revealed the presence of
alkaloids, flavonoids, tannins, phytosterols, saponins, glycosides, quinones and resins. The present work justifies the use of Urginea
indica by tribal community of Bastar in combating several ailments.
Keywords: Activity index, Antimicrobial activity, Extracts, Percentage yield, Phytochemical analysis, Solvents, Urginea indica.
INTRODUCTION
M
edicinal plants are the richest bio-resources for
the discovery of novel bioactive compounds.
They are gifts of nature to cure several diseases
among human beings.1 Their curative potentiality is due
to the presence of a wide array of complex chemical
compounds known as secondary metabolites viz.,
alkaloids, flavonoids, saponins and phenolic compounds
present in different parts namely root, stem and leaf
which are used for antimicrobial and therapeutic
purposes.2,3 There are several reports documenting the
efficacy of plant extracts on microorganism by a number
of researchers in different parts of the world.4,5 Although
many of the plant species have been tested for their
antimicrobial efficacy still the vast majority has not been
6,7
evaluated thus far. It is, therefore, essential for the
systematic evaluation and scientific validation of plants
used in traditional medicine for several diseases. Hence,
there is an urgent need to screen medicinal plants for
exploring their promising biological activity.
Herbal medicines are in great demand in the developed
as well as developing countries due to their wide
medicinal and biological applications. The healing
potentiality of the plant continues to play significant role
in the primary health care of about 80% of the world’s
population.8 The uses of plant extracts as antimicrobials
have become popular because the effective life span of
antibiotic is limited and over prescription as well as
misuse of antibiotics are causing antimicrobial
resistance.9 The increased prevalence of multidrug
resistance in pathogenic microorganisms and undesirable
side effects of certain antibiotics have triggered immense
interest in the search for new antimicrobial drugs of plant
origin.
Urginea indica commonly called as wild onion is endemic
to India, Africa and Mediterranean regions. It is found in a
wide habitats ranging from desert, shrub, grassland,
dunes and forests. It is a perennial geophyte with fibrous
roots of six to ten inches of length, The rounded conical,
pear shaped bulbs with white transparent outer scales
are about the size of an big onion, consisting of fleshy
coats which are thin and papery red or orange brown in
colour enclosing each other completely. Indian squill is
considered to have medicinal value and is largely used as
an expectorant, cardiac stimulant, in treating
rheumatism, dropsy, edema, gout, asthma and as an
anticancer agent. Squill bulbs have long been used as a
source of natural product with pharmaceutical and
biocidal applications.10 Chhattisgarh state is identified as
an herbal state and has rich resource of medicinal plants
in many tribal districts including Bastar. Not much work
has been done in this ectopic zone on the antimicrobial
properties of plants. Thus, in this context the present
study was carried out to screen the antimicrobial efficacy
and phytochemical analysis of Urgenia indica against both
gram positive and gram negative bacteria viz., Bacillus
cerus, Bacillus subtilis, Staphylococcus aureus,
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Int. J. Pharm. Sci. Rev. Res., 26(2), May – Jun 2014; Article No. 47, Pages: 273-281
Staphylococcus epidermidis, Escherichia coli, Proteus
vulgaris, Pseudomonas aeruginosa and Klebsiella
pneumoneae and against two fungi viz., Aspergillus niger
and Candida albicans.
MATERIALS AND METHODS
Selection of medicinal plant
Plant of Urginea indica was selected based on its
traditional usage by the tribal community of Bastar
district of Chhattisgarh in curing several ailments and its
ethno-medicinal importance as herbal drug. Apparently
healthy and diseased free plants were selected for the
assessment of antimicrobial activity and phytochemical
analysis.
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Collection of the sample
The fresh and healthy root, stem and leaves of Urginea
indica were collected and identified at Department of
Horticulture, Shahid Gundadhoor College of Agriculture
and Research Station (SGCARS), Kumhrawand, Jagdalpur
from Bastar district, Chhattisgarh, India. The plant
samples were washed under running tap water to remove
debris and shade dried for about three weeks to attain a
constant weight. The dried samples were mechanically
grinded by using a mortar and pestle and finally
powdered by laboratory blender (Remi) and stored in
separate air tight bottles till use (Figure 1).
Root
Leaf
Stem
Figure 1: Sample collection of different parts of Urginea indica
Table 1: Physical characteristics of the extract of Urginea indica in different solvents
Plant Parts
Root Extract
Stem Extract
Leaf Extract
Characteristics
Different Solvents used for extraction
Chloroform
Acetone
Methanol
Aqueous
Percent Yield
1.53
2.73
5.13
6.40
Colour
Yellowish
Reddish brown
Brown
Pale yellow
Consistency
Sticky
Sticky
Viscous
Sticky
Percent Yield
2.46
1.00
5.03
4.26
Colour
Yellow
Redish
Red
Redish brown
Consistency
Waxy
Viscous
Sticky
Viscous
Percent Yield
4.86
0.53
2.86
3.20
Colour
Yellowish green
Greenish
Greenish
Dark brown
Consistency
Viscous
Waxy
Semisolid
Solid
Extraction procedure
15g powdered material was extracted with 150 ml of
different solvents according to their increasing polarity
successively for 8-10 hours in the soxhlet apparatus
(Tempo) at a temperature not exceeding the boiling point
of the respective solvents. After extraction excess solvent
was removed by distillation and the concentrated extracts
so obtained were further dried in incubator at 40oC. The
percentage yield and other physical properties were
recorded. The residual extracts after drying were
dissolved in 50% dimethyl sulphoxide and stored in
refrigerator at 4oC in small and sterile glass tubes.
Microorganisms used for the test
The present study was carried out with the microbial
strains procured from IMTECH, Chandigarh, India. The
microbial strains used for antimicrobial assessment were
Bacillus cereus (MTCC-430), Bacillus subtilis (MTCC-441),
Staphylococcus aureus (MTCC-96), Staphylococcus
epidermidis (MTCC-435), Escherichia coli (MTCC-1687),
Proteus vulgaris (MTCC-744), Pseudomonas aeruginosa
(MTCC-741), Klebsiella Pneumoniae (MTCC-3384)
Aspergillus niger (MTCC-872) and Candida albicans
(MTCC-183). The bacterial and fungal strains were
maintained on nutrient agar and potato dextrose agar
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Int. J. Pharm. Sci. Rev. Res., 26(2), May – Jun 2014; Article No. 47, Pages: 273-281
ISSN 0976 – 044X
slants respectively, sub cultured regularly and stored at
o
4 C for further use.
the standardized cell suspension were spread on Muller
Hinton Agar (Hi-media) plate using a sterile swab and air
dried to remove the surface moisture. Wells were bored
Inoculum preparation
into the agar plate using a sterile 6 mm diameter cork
One loop full of overnight grown bacterial culture was
borer. The crude extract was introduced into the well at a
o
inoculated in 25 ml nutrient broth at 37 C on a rotary
concentration of 2mg/20µl, allowed to stand at room
shaker incubator for 16-18 h. The inoculum size of each
temperature for about 1 h as a period of pre-incubation
bacterial strains were standardized by adjusting the
diffusion to minimize the effect of variation in time
optical density of the culture broth to a turbidity
between the application of different solutions and later
corresponding to 0.08 at 620 nm using a
the plates were incubated at 37oC for 24 h. Controls were
8
11
spectrophotometer which is equivalent to 10 cfu/ml.
also set up in parallel and the effects were compared with
penicillin and streptomycin at a concentration of
Antibacterial activity
10µg/20µl. The plates were observed for the zone of
The antibacterial activity of the crude extracts was
inhibition after 24 h. The experiment was conducted in
determined by the agar-well diffusion method.12 200 µl of
triplicates and the values are expressed as mean ± SE.
Table 2: Antibacterial activity (Zone of inhibition in mm, Mean ± SE) of Urginea indica
Bacteria
Chloroform
Different Solvents for extraction
Acetone
Methanol
Aqueous
Standard antibiotics
P*
S**
Root
B. cerus
B. subtilis
11.80±0.20
-
14.13±0.13
-
15.06±0.06
-
-
11.33±0.17
09.33±0.24
25.13±0.06
20.80±0.11
S. aureus
S. epidermidis
E. coli
P. vulgaris
P. aeruginosa
10.06±0.06
11.40±0.11
14.20±0.20
11.20±0.11
12.40±0.23
16.26±0.26
12.33±0.17
14.33±0.24
13.13±0.13
-
25.40±0.23
28.26±0.13
08.40±0.20
-
26.60±0.20
24.73±0.26
18.93±0.06
15.53±0.24
13.93±0.06
K. pneumoneae
Stem
B. cerus
B. subtilis
S. aureus
12.46±0.24
13.40±0.23
10.40±0.11
-
-
17.86±0.13
11.13±0.13
09.06±0.06
12.20±0.20
10.20±0.11
14.06±0.06
11.46±0.24
-
11.33±0.17
09.33±0.24
25.40±0.23
25.13±0.06
20.80±0.11
26.60±0.20
S. epidermidis
E. coli
P. vulgaris
P. aeruginosa
K. pneumoneae
Leaf
09.60±0.11
12.33±0.17
11.20±0.20
10.46±0.13
14.06±0.06
12.40±0.23
13.53±0.17
12.26±0.26
09.40±0.23
-
28.26±0.13
08.40±0.20
-
24.73±0.26
18.93±0.06
15.53±0.24
13.93±0.06
17.86±0.13
B. cerus
B. subtilis
S. aureus
S. epidermidis
E. coli
08.13±0.13
08.46±0.24
-
09.26±0.26
09.53±0.13
-
10.06±0.06
11.20±0.20
-
-
11.33±0.17
09.33±0.24
25.40±0.23
28.26±0.13
08.40±0.20
25.13±0.06
20.80±0.11
26.60±0.20
24.73±0.26
18.93±0.06
P. vulgaris
P. aeruginosa
K. pneumoneae
10.33±0.24
-
12.33±0.17
-
09.20±0.11
-
-
-
15.53±0.24
13.93±0.06
17.86±0.13
Standard antibiotics: P*- (Penicillin) and S**- (Streptomycin); (-) No zone detected
Antifungal activity
The antifungal activity of the plant extract was evaluated
13
by agar-well diffusion method. potato dextrose agar was
used to culture the fungal species. The spore suspension
was spread on potato dextrose agar (Hi-media) plate
using a sterile swab and air dried to remove the surface
moisture. Wells were bored into the agar using a sterile 6
mm diameter cork borer. The crude extract was
aseptically introduced into the well at a concentration of
2mg/20µl, allowed to stand at room temperature for
about 1 h as a period of pre-incubation diffusion to
minimize the effect of variation in time between the
application of different solutions and later the plates
were incubated at 28-30oC for 48 h. Controls were also
set up in parallel and the effects were compared with
clotrimazole as standard antifungal at a concentration of
10µg/20µl. The plates were observed for the zone of
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Int. J. Pharm. Sci. Rev. Res., 26(2), May – Jun 2014; Article No. 47, Pages: 273-281
inhibition after 48 h. The experiment was conducted in
triplicates and the values are expressed as mean ± SE.
Qualitative phytochemical analysis
Phytochemical analysis of the extracts of root, stem and
14,15
leaf of Urginea indica were performed.
Test for Alkaloids
(a) Mayer’s Test: To 1ml of extract add 1ml of Mayer’s
reagent (Potassium mercuric iodide solution) whitish
yellow or cream colored precipitate indicated the
presence of alkaloids.
(b) Hager’s Test: To 1ml of the extract, add 1ml of Hager’s
reagent (Saturated aqueous solution of picric acid). A
yellow colored precipitate indicates the presence of
alkaloids.
(c) Wagner’s Test: To 1 ml of the extract, add 1 ml of
Wagner’s reagent (Iodine in potassium iodide solution).
Formation of reddish brown precipitate indicates the
presence of alkaloids.
Test for Flavonoids
(a) Alkaline reagent Test: Test solution when treated with
NaOH solution shows increase in intensity of yellow
colour, which becomes colorless on addition of few drops
of dilute acid.
(b) Lead acetate solution Test: Test solution with few
drops of lead acetate solution (10%) gives yellow
precipitate.
Test for Saponins
(a) Foam Test: The extract was shaken vigorously with
water. Persistent foam was observed.
Test for Tannins
(a) Gelatin Test: To the extract added 1% gelatin solution
in water and appearance of white precipitate was
observed.
(b) Feric chloride Test: To the extract added 5% ferric
chloride solution and deep blue-black colour was
observed.
Test for phytosterols
Small quantity of extract is dissolved in 5 ml of chloroform
separately. The above obtained chloroform solution are
subjected to the following tests.
(a) Salkowski Test: To the 1ml above prepared chloroform
solution few drops of conc. H2SO4 is added. Formation of
brown ring indicates the presence of phytosterols.
(b) Libermann-Burchard Test: The above prepared
chloroform solution are treated with few drops of conc
H2SO4 followed by 1 ml of acetic anhydride solution. A
bluish green colour solution shows the presence of
phytosterols.
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Test for Glycosides
10 ml of 50% H2SO4 was added to 1ml of each extract in
separate test tubes. The mixture was heated in a boiling
water bath for 15 minutes and 10ml (5ml each of Fehling
A and Fehling B) was added to the mixture and boiled.
The presence of brick red precipitate indicates the
presence of glycosides.
Test for Quinones
A few drops of 1% NaOH was mixed with the plant extract
and shaken vigorously. A blue green or red colour
indicates the presence of quinones.
Test for Resins
To the extract in a test tube, 5ml of the 1% copper
acetate solution was added and the resulting solution was
shaken vigorously and allowed to separate. The
separation of a green colored solution was considered as
positive for resins.
RESULTS AND DISCUSSION
Natural therapeutic products play a vital role in the
development of novel drugs for the treatment of several
diseases.16 In the present investigation the plant samples
were successively extracted with the soxhlet apparatus
using four different solvents based on their polarity index
as chloroform, acetone, methanol and aqueous to ensure
that a wide polarity range of compounds could be
extracted.17 The different parts of Urginea indica viz.,
root, stem and leaf were assessed for their antimicrobial
efficacy as the antimicrobial phytochemical compounds of
the plant origin may occur mostly in root, stem and leaf of
the plants.18,19
The physical properties of the selected different parts of
Urginea indica viz., root, stem and leaf extracts were
assessed and the result revealed that the root extracts
were almost sticky to viscous in nature with yellowish to
reddish brown in colour and the percentage yield was
found to be highest in root extracts in case of polar
aqueous solvent (6.40%) followed by methanol (5.13%),
acetone (2.73%) and chloroform (1.53%) respectively. The
highest percentage yield in aqueous solvent might be due
to the fact that water is a universal solvent and extracts
20,21
most of the compounds.
The stem extracts were
yellow to red in colour and leaf extracts were almost
viscous to semisolid in nature with yellowish green to
dark brown in colour and the percentage yield of the
stem and leaf extracts were found to be maximum in case
of polar organic solvents (5.03%) and non polar solvents
(4.86%) respectively (Table 1). The findings of the above
study clearly illustrate the presence of varied
phytochemical in different parts of the plants and their
affinity towards different solvents according to their
polarity index.
The antimicrobial activity in crude extract of root, stem
and leaf of Urginea indica were assessed against both
gram positive and gram negative bacteria viz., Bacillus
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Int. J. Pharm. Sci. Rev. Res., 26(2), May – Jun 2014; Article No. 47, Pages: 273-281
cerus, Bacillus subtilis, Staphylococcus aureus,
Staphylococcus epidermidis, Escherichia coli, Proteus
vulgaris Pseudomonas aeruginosa and Klebsiella
pneumoneae. The study revealed that the root extracts
were found to be inhibitory to these bacteria followed by
stem and leaf. The antibacterial activity in case of gram
positive bacteria was found to be high in methanolic root
extracts followed by acetone and chloroform. The
methanolic root extract exhibited significant zone of
inhibition against three gram positive bacteria with
maximum against Bacillus cerus (15.06±0.06) followed by
Staphylococcus epidermidis (14.33±0.24), Staphylococcus
aureus (12.33±0.17). The stem and leaf extract showed
activity against Bacillus cerus followed by Staphylococcus
epidermidis and Staphylococcus aureus, whereas no
significant activity was observed in case of their aqueous
extract against all the bacterial species tested. These
results clearly indicate that organic solvents were more
suitable for the extraction of the active principles
responsible for antibacterial activity.22-24 The decline in
activity of aqueous extract might be due to the excessive
heating of the aqueous soluble active constituents during
the extraction process which often affects the activity of
the phytoconstituents present in the extract.25-27 The
antibacterial activity in case of gram negative bacteria
was found to be comparatively less than that of gram
positive bacteria against all the extracts tested. The
acetone extract of root exhibited maximum zone of
inhibition against Pseudomonas aeruginosa (16.26±0.26)
followed by Klebsiella pneumoneae (13.40±0.23) except
for Proteus vulgaris and Escherichia coli. Amongst gram
negative bacteria, Pseudomonas aeruginosa showed the
highest inhibition whereas Proteus vulgaris and
Escherichia coli were found to be resistant against all the
extracts tested. In the present investigation the gram
positive bacteria were found to be more susceptible as
compared to gram negative bacteria against the plant
extracts tested. The higher resistance of gram-negative
bacteria to plant extracts is due the presence of outer
phospholipids membrane carrying the structural
lipopolysaccharides components, which makes the cell
wall impermeable to antimicrobial substances whereas,
gram positive bacteria have single layered cell wall with
peptidoglycan constituting the outer layer, which is not
an effective permeability barrier.28,29 The potential
sensitivity of the extracts against gram positive and gram
negative bacteria and the zone of inhibition were
recorded and are presented in (Table 2, Figure 2).
The antifungal activity was assessed against two
pathogenic fungi viz., Aspergillus niger and Candida
albicans. The results revealed that the acetone extract of
root was effective against both the fungal species with
maximum zone of inhibition against Candida albicans
(14.06±0.06) and Aspergillus niger (13.26± 0.26) followed
by stem and leaf extracts whereas aqueous and
chloroform extracts were not found to be inhibitory
against these two fungal species (Table3, Fig.3). The
antifungal activity of the studied plant was compared
ISSN 0976 – 044X
with standard antifungal a similar study for the
assessment of plant extracts against different fungal
pathogens is also well documented.30,31
(a) Pseudomonas aeruginosa
(b) Bacillus cerus
Figure 2: Antibacterial activity of U. indica against
Pseudomonas aeruginosa and Bacillus cerus
Table 3: Antifungal activity of Urginea indica (Zone of
inhibition in mm, Mean ± SE)
Fungal Species
Root
Chloroform
Acetone
Methanol
A.niger
C.albicans
13.26±0.26
09.80±0.20
14.06±0.06
11.13±0.13
Aqueous
Clotrimazole
Stem
Chloroform
Acetone
Methanol
14.33±0.33
7.66±0.06
11.06±0.06
09.20±0.20
11.53±0.26
10.46±0.17
Aqueous
Clotrimazole
Leaf
Chloroform
Acetone
14.33±0.33
7.66±0.06
08.20±0.20
09.20±0.11
Methanol
Aqueous
Clotrimazole
07.06±0.06
14.33±0.33
07.46±0.24
7.66±0.06
Standard antifungal: Clotimazole; (-) No zone detected
(a) Candida albicans
(b) Aspergillus niger
Figure 3: Antifungal activity of U. indica against Candida
albicans and Aspergillus niger
The activity index was calculated to express the
relationship between zones of inhibition of the extracts
with the standard antibiotics.32 Among the root, stem and
leaf extracts of U. indica, the highest activity index of 0.59
was recorded in gram positive bacteria for B. cerus
whereas, activity index of 1.16 was observed for gram
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Int. J. Pharm. Sci. Rev. Res., 26(2), May – Jun 2014; Article No. 47, Pages: 273-281
negative bacteria as P. aeruginosa against streptomycin
(Table 4). The antifungal activity showed highest activity
index in case of acetone extracts of root in case of
Candida albicans (1.83) followed by Aspergillus niger
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(0.92) (Table 5). Higher activity index (>0.5) in the crude
extract indicates potential antimicrobial activity in the
plant.
Table 4: Activity index of the extract of Urginea indica in different solvents with respect to Streptomycin
Different Solvents used for extraction
Bacterial Species
B. cerus
B. subtilis
S. aureus
Root
0.46
0.37
Chloroform
Stem
0.44
0.34
Leaf
0.30
Root
0.56
0.42
Acetone
Stem
0.48
0.38
Leaf
0.34
Root
0.59
0.46
Methanol
Stem
0.55
0.43
Leaf
0.37
S. epidermidis
E. coli
P. vulgaris
P. aeruginosa
K. pneumoneae
0.46
1.01
0.69
0.38
0.88
0.62
0.34
0.74
-
0.50
1.16
0.75
0.42
1.00
0.69
0.38
0.88
-
0.57
0.94
0.58
0.54
0.88
0.52
0.45
0.66
-
Table 5: Activity index of the extract of Urginea indica in
different solvents with respect to Clotrimazole
Phytochemical constituents such as alkaloids, flavonoids,
tannins, phenols, saponins, and several other aromatic
compounds are secondary metabolites of plants that
serve as defense against prediction by many
microorganisms, insects and other herbivores.33 The
present study revealed the presence of medicinally active
phytoconstituents in different parts of the plants viz.,
root, stem and leaf. The phytochemical analysis of
Urginea indica extracts are summarized in (Tables 6-8).
Different Solvents used for extraction
Fungal
Species
Acetone
Methanol
Root
Stem
Leaf
Root
Stem
Leaf
A. niger
0.92
0.77
0.57
0.68
0.62
0.49
C.
albicans
1.83
1.50
1.20
1.45
1.36
0.97
Table 6: Phytochemical analysis from the root extract of Urginea indica in different solvents
Phytochemical Test
Chloroform
Acetone
Methanol
Aqueous
Mayer’s test
-
+
-
-
Wagner’s test
-
+
-
-
Hager’s test
-
+
-
-
Alkaline reagent test
+
+++
++
+
Lead acetate test
+
+++
++
+
Ferric chloride test
-
++
-
-
Gelatin test
-
++
-
-
Salkowski test
++
+++
++
+
Libermann-Burchard test
++
+++
++
+
-
+
-
-
Glycosides
-
+++
++
+
Quinones
-
+
-
-
+++
++
-
-
Alkaloids
Flavonoids
Tannins
Phytosterols
Saponins
Foam test
Resins
“+++” Extremely Positive; “++” Strongly Positive; “+” Positive; “-” Negative
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Int. J. Pharm. Sci. Rev. Res., 26(2), May – Jun 2014; Article No. 47, Pages: 273-281
ISSN 0976 – 044X
Table 7: Phytochemical analysis from the stem extract of Urginea indica in different solvents
Phytochemical Test
Chloroform
Acetone
Methanol
Aqueous
Alkaloids
Mayer’s test
-
+
-
-
Wagner’s test
-
+
-
-
Hager’s test
-
+
-
-
Alkaline reagent test
-
+++
++
+
Lead acetate test
-
+++
++
+
Ferric chloride test
-
++
-
-
Gelatin test
-
++
-
-
Salkowski test
+
+++
++
++
Libermann-Burchard test
+
+++
++
++
Foam test
-
+
-
-
Glycosides
-
+++
++
+
Quinones
-
-
-
-
+++
++
-
-
Flavonoids
Tannins
Phytosterols
Saponins
Resins
“+++” Extremely Positive; “++” Strongly Positive; “+” Positive; “-” Negative
Table 8: Phytochemical analysis from the leaf extract of Urginea indica in different solvents
Phytochemical Test
Chloroform
Acetone
Methanol
Aqueous
Mayer’s test
-
-
-
-
Wagner’s test
-
-
-
-
Hager’s test
-
-
-
-
Alkaline reagent test
-
+
+
+
Lead acetate test
-
+
+
+
Ferric chloride test
-
-
-
-
Gelatin test
-
-
-
-
Salkowski test
+
++
+
+
Libermann-Burchard test
+
++
+
+
Foam test
-
-
-
++
Glycosides
-
-
-
+
Quinones
-
-
-
-
-
-
Alkaloids
Flavonoids
Tannins
Phytosterols
Saponins
Resins
+++
-
“+++” Extremely Positive; “++” Strongly Positive; “+” Positive; “-” Negative
Analysis of plant extracts revealed the presence of
alkaloids, flavonoids, tannins, saponins, phytosterols,
qinones, glycosides and resins in different parts of the
plant. Similar findings were also reported and well
documented.34,35 The phytochemical studies have shown
that plants with antimicrobial activity mainly contain
bioactive constituents such as alkaloids, flavonoids,
tannins and saponins.36 Alkaloids are the largest group of
phytochemicals causing toxicity against cells of foreign
organisms.37 Saponin possess antimicrobial property due
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279
Int. J. Pharm. Sci. Rev. Res., 26(2), May – Jun 2014; Article No. 47, Pages: 273-281
to its ability to cause leakage of certain enzymes and
proteins from the cell.38 Flavonoids are hydroxylated
phenolic substances synthesized by the plants in response
to microbial infection and has the ability to complex with
extracellular and soluble proteins and to complex with
bacterial cell walls.39 Tannins are known to bind with
proline rich proteins and interfere in the protein
synthesis.40 Steroids have been documented to have
antibacterial properties, the steroids specifically associate
with membrane lipid and exerts its action by causing
leakages from liposomes.41 The findings of the present
study offers a scientific validation for the usage of
Urginea indica in curing several ailments by the
traditional healers.
4.
Bonjar GHS, Nik AK, Aghighi S, Antibacterial and antifungal
survey in plants used in indigenous herbal-medicine of
south east regions of Iran, Journal of Biological Sciences, 4,
2004, 405-412.
5.
Boer HJ, Kool A, Broberg A, Mziray WR, Hedberg I,
Levenfors JJ, Antifungal and antibacterial activity of some
herbal
remedies
from
Tanzania,
Journal
of
Ethnopharmacology, 96, 2005, 461-469.
6.
Tshikalange TE, Meyer JJM, Hussein AA, Antimicrobial
activity, toxicity and the isolation of a bioactive compound
from plants used to treat sexually transmitted diseases,
Journal of Ethnopharmacology, 96, 2005, 515-519.
7.
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TS, Isolation and identification of bioactive compounds
responsible for antibacterial efficacy of Lotus corniculatus,
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different organic extracts of Achyranthes aspera and Cassia
alata, Journal of Scientific Research, 1, 2009, 393-398.
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Shiva Kameshwari MN, Thara Sarawathi KJ, Muniyamma M,
Morphological Variations in population of Urginea indica,
Kunth. Liliaceae, Journal of Natural and Applied Science, 2,
2010, 280 – 289.
11.
Basri DF, Fan SH, The potential of aqueous and acetone
extracts of galls of Quercus infectoria as antibacterial
agents, Indian Journal of Pharmacology, 37, 2005, 26-29.
12.
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extracts of Racarpu villosus, Journal of Ethnopharmacology,
42, 1994, 39-43.
13.
Gobdi TA, Irobi ON, Antibacterial activities of crude extracts
of Aspergillus quadrilineatus isolated from Nigerian cereal,
African Journal of Pharmacy and Pharmaceutical Sciences,
22, 1992, 101-106.
14.
Harborne JB, A Guide to Modern Techniques of Plant
rd
Analysis, 3 edition, Chapman and Hall International, New
York, 1998.
15.
Kokate CK, Pharmacognosy, 16 edition, Nirali Prakashan,
Mumbai, India, 2001.
16.
Newman DJ, Cragg CM, Snadder KM, Natural products as
sources of new drugs over the period, Journal of Natural
Product, 66, 2003, 1022-1037.
17.
Pandey D, Gupta AK, Screening of the antibacterial activity
of Amorphophallus campanulatus from Bastar region of
Chhattisgarh, India, International Journal of Applied Biology
and Pharmaceutical Technology, 4, 2013, 1-6.
18.
Siddiqui S, Verma A, Rather AA, Jabeen F, Meghvansi MK,
Preliminary phytochemicals analysis of some important
medicinal and aromatic plants, Advances in Biological
Research, 3, 2009, 188-195.
19.
Manivannan R, Mannangatti VK, Ayyasamy B, Natesan SK,
Pharmacognostical and preliminary phytochemical studies
of Bauhinia tomentosa L., Journal of Pharmacy Research, 3,
2010, 502-505.
CONCLUSION
The present investigation was focused on the
antimicrobial activity and phytochemical analysis in the
root, stem and leaf extracts of Urginea indica in four
different solvents against four gram positive, four gram
negative bacteria and two fungi. The results were
promising and revealed that The highest activity index
was recorded in root methanol extracts in Bacillus cerus
and acetone extracts in case of Pseudomonas aeruginosa
respectively, whereas root acetone extract exhibited
pronounced activity against Aspergillus niger and Candida
albicans. Since, medicinal plants are considered as
clinically effective and safer alternatives to the synthetic
antibiotics. Thus there is an urgent need of extensive
research in the area of medicinal plants with the aim of
exploring their ethno-medicinal use and subsequently the
isolation and characterization of bioactive compounds will
contribute for the better, safer and cost effective noval
drug development.
Acknowledgements: The authors are thankful to the
authorities of Pt. Ravishankar Shukla University, Raipur
(C.G.) for providing research facilities. The authors
acknowledge DST, New Delhi for providing FIST grant to
School of Life Sciences. MTCC cultures procured from
IMTECH, Chandigarh and plant samples collection and
identification at (SGCARS) Kumhrawand, Jagdalpur, (C.G.)
is duly acknowledged. One of the author (DP) is grateful
to Department of Science and Technology, New Delhi for
the financial assistance for research in the form of
INSPIRE Fellow-SRF.
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Source of Support: Nil, Conflict of Interest: None.
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