Content uploaded by Chandra Mohini Nemkul
Author content
All content in this area was uploaded by Chandra Mohini Nemkul on Feb 17, 2020
Content may be subject to copyright.
98
2019 Journal of Plant Resources Vol.17, No. 1
Phytochemical Evaluation and In Vitro Antimicrobial Activity of the
Roots of Flemingia strobilifera (L.) R. Br.
Chandra Mohini Nemkul1*, Gan B. Bajracharya2 and Ila Shrestha3
1Tri-chandra Multiple Campus, Tribhuvan University, Kathmandu, Nepal
2Nepal Academy of Science and Technology (NAST), Khumaltar, Lalitpur, Nepal
3Patan Multiple Campus, Tribhuvan University, Patan Dhoka, Lalitpur, Nepal
*Email: Chandra.mohini21@gmail.com
Abstract
Ethno-medicinal uses of Flemingia strobiifera in the Magar communities at Kawaswoti urban
municipality, Province no. 4, Nepal was surveyed. Phytochemicals present in the hexane and
aqueous methanolic extracts of the roots were evaluated by chemical tests and GC-MS analysis.
The antimicrobial activity of the extracts was carried out against 8 bacterial species by the agar
well diffusion method. Zone of inhibition was compared with standard antibiotics ampicillin and
gentamicin. The aqueous methanolic extract showed stronger antimicrobial activity against
Escherichia coli. The lowest MIC and MBC values were 1.56 and 6.25 mg/ml, respectively.
Phytochemical screening revealed the presence of polyphenols and terpenoids. The antimicrobial
activity of the plant material might be due to the presence of these phytochemicals.
Keywords: Ethno-medicine, Magar community, Phytoconstituents, Zone of inhibition
Introduction
Flemingia strobilifera (L.) R. Br. belongs to family
Leguminosae. It is known as bharkauli jhar and
bhatwasi in Nepali. It is used in folkloric medicine,
such as leaves and flowers for tuberculosis, and roots
for ulcers, body swellings, epilepsy, insomnia, fever,
indigestion, diarrhea and dysentery (Bhattarai, 1991;
Manandhar, 2002; Ghalot et al., 2011; Kumar et al.,
2011b). It is used as fodder by Chepang communities
in mid hills of Nepal (Rijal, 2011). Root powder is
applied on the body by Darai tribe of Chitwan
district, Nepal for scabies (Dangol & Gurung, 2000).
Madan et al. (2009) have isolated isoflavonoids from
F. strobilifera roots and showed antimicrobial
activity against Staphylococcus aureus, Pseudomonas
aeruginosa, Methicillin-resistant Staphylococcus
aureus and Escherichia coli. Kumar et al. (2011a)
reported a significant anthelmintic activity of the
alcoholic and chloroform extracts of the leaves of F.
strobilifera. Roots of F. strobilifera constituted
phenols, flavonoids, steroids, flavonoids glycosides
and tannins (Madan et al., 2010).
From the field studies, it was came to know that the
Magar communities in Kawaswoti urban
municipality, Province no. 4, Nepal use juice of F.
strobilifera roots for the treatment of diarrhea,
dysentery and gastritis. Therefore to validate ethno-
medicinal knowledge, antimicrobial susceptibility
test of F. strobilifera root extracts was evaluated in
the present work.
Materials and Methods
Field visit
The study was carried out in Kawaswoti urban
municipality of Nawalpur district, Province no. 4,
Nepal. Ethno-medicinal data of the medicinal plants
of the Magar communities were collected during
field visit in April, 2016. Herbaria were prepared
and confirmed through comparison with specimens
at National Herbarium and Plant Laboratories,
Godawari, Nepal.
Materials
Mueller Hinton Agar (MHA) and Mueller Hinton
Broth (MHB) were purchased from HiMedia
Laboratories Pvt. Ltd. Hexane and Methanols were
purchased from Fisher Scientific.
J. Pl. Res. Vol. 17, No. 1, pp 98-103, 2019
98
2019 Journal of Plant Resources Vol.17, No. 1
Phytochemical Evaluation and In Vitro Antimicrobial Activity of the
Roots of Flemingia strobilifera (L.) R. Br.
Chandra Mohini Nemkul1*, Gan B. Bajracharya2 and Ila Shrestha3
1Tri-chandra Multiple Campus, Tribhuvan University, Kathmandu, Nepal
2Nepal Academy of Science and Technology (NAST), Khumaltar, Lalitpur, Nepal
3Patan Multiple Campus, Tribhuvan University, Patan Dhoka, Lalitpur, Nepal
*Email: Chandra.mohini21@gmail.com
Abstract
Ethno-medicinal uses of Flemingia strobiifera in the Magar communities at Kawaswoti urban
municipality, Province no. 4, Nepal was surveyed. Phytochemicals present in the hexane and
aqueous methanolic extracts of the roots were evaluated by chemical tests and GC-MS analysis.
The antimicrobial activity of the extracts was carried out against 8 bacterial species by the agar
well diffusion method. Zone of inhibition was compared with standard antibiotics ampicillin and
gentamicin. The aqueous methanolic extract showed stronger antimicrobial activity against
Escherichia coli. The lowest MIC and MBC values were 1.56 and 6.25 mg/ml, respectively.
Phytochemical screening revealed the presence of polyphenols and terpenoids. The antimicrobial
activity of the plant material might be due to the presence of these phytochemicals.
Keywords: Ethno-medicine, Magar community, Phytoconstituents, Zone of inhibition
Introduction
Flemingia strobilifera (L.) R. Br. belongs to family
Leguminosae. It is known as bharkauli jhar and
bhatwasi in Nepali. It is used in folkloric medicine,
such as leaves and flowers for tuberculosis, and roots
for ulcers, body swellings, epilepsy, insomnia, fever,
indigestion, diarrhea and dysentery (Bhattarai, 1991;
Manandhar, 2002; Ghalot et al., 2011; Kumar et al.,
2011b). It is used as fodder by Chepang communities
in mid hills of Nepal (Rijal, 2011). Root powder is
applied on the body by Darai tribe of Chitwan
district, Nepal for scabies (Dangol & Gurung, 2000).
Madan et al. (2009) have isolated isoflavonoids from
F. strobilifera roots and showed antimicrobial
activity against Staphylococcus aureus, Pseudomonas
aeruginosa, Methicillin-resistant Staphylococcus
aureus and Escherichia coli. Kumar et al. (2011a)
reported a significant anthelmintic activity of the
alcoholic and chloroform extracts of the leaves of F.
strobilifera. Roots of F. strobilifera constituted
phenols, flavonoids, steroids, flavonoids glycosides
and tannins (Madan et al., 2010).
From the field studies, it was came to know that the
Magar communities in Kawaswoti urban
municipality, Province no. 4, Nepal use juice of F.
strobilifera roots for the treatment of diarrhea,
dysentery and gastritis. Therefore to validate ethno-
medicinal knowledge, antimicrobial susceptibility
test of F. strobilifera root extracts was evaluated in
the present work.
Materials and Methods
Field visit
The study was carried out in Kawaswoti urban
municipality of Nawalpur district, Province no. 4,
Nepal. Ethno-medicinal data of the medicinal plants
of the Magar communities were collected during
field visit in April, 2016. Herbaria were prepared
and confirmed through comparison with specimens
at National Herbarium and Plant Laboratories,
Godawari, Nepal.
Materials
Mueller Hinton Agar (MHA) and Mueller Hinton
Broth (MHB) were purchased from HiMedia
Laboratories Pvt. Ltd. Hexane and Methanols were
purchased from Fisher Scientific.
J. Pl. Res. Vol. 17, No. 1, pp 98-103, 2019
99
2019 Journal of Plant Resources Vol.17, No. 1
Preparation of the plant extracts
Roots of F. strobilifera were dried in shade at room
temperature. Air dried plant materials were ground.
The ground plant material (100 g) was successively
extracted with hexane (800 ml, 7 hours) and 70%
methanol (800 ml, 22 hours) using a Soxhlet
extractor. These plant extracts were concentrated by
using a rotary evaporator and vacuum dried. The
extracts were stored in a refrigerator at 4ºC until
further use.
Phytochemical screening
Phytochemical screening of the hexane and aq.
methanolic extracts was performed using different
specific reagents to find out different
phytoconstituents present in the plant extracts
(Ciulei, 1982). Among other tests, Braymer,
Dragendorff, Liebermann-Burchard and Salkowski
tests were carried out to detect polyphenols, alkaloid,
steroids and terpenoids, respectively.
Gas chromatography-mass spectrometry (GC-MS)
GC-MS analyses of the hexane and aq. methanolic
extracts of F. strobilifera was analyzed using an
Agilent 7890A GC system coupled with an Agilent
5975 C mass selective detector, equipped with a HP-
5MS GC column (5% phenyl methyl siloxane,
Agilent 19091S-433, 30 m × 250 µm internal
diameter, 0.25 µm film thickness). Helium was used
as a carrier gas at flow rate of 1.21 ml/min. The
instrument was operated in the electron impact (EI)
mode at 70 eV and ion source temperature 230°C in
the scan range of 50-500 m/z. The initial column
temperature was set at 40°C held for 2 min, ramped
at a rate of 4°C/min to 270°C and held for 5.5 min
(total run time 65 min). A dilute sample solutions of
the extracts were prepared in HPLC grade hexane
and methanol, and a volume of 2 µl was injected.
The constituents were identified by comparing the
mass spectra available in a MS database (NIST 08).
Antimicrobial susceptibility test
The hexane and aq. methanolic extracts were
screened against a total of 8 bacterial strains namely
Pseudomonas aeruginosa (ATCC 27263),
Staphylococcus aureus (ATCC 25923), Escherichia
coli (ATCC 25922), Klebsiella pneumoniae ( ATCC
700603), Enterococcus faecalis (ATCC 29212),
Bacillus subtilis (ATCC 6051), Shigella dysenteriae
(ATCC 13313) and Salmonella enteric subsp. enteric
serovar typhi.
Inoculums were prepared to McFarland standard 0.5
as described in Nemkul et al., (2018). The inoculums
were used within 30 minutes.
The antibacterial screening of these extracts was
evaluated by using the agar well diffusion technique
(Perez et al., 1990). The standardized bacterial
inoculums were uniformly spread on the respective
sterile MHA agar Petri dishes using sterile cotton
swabs. The wells were punched on the agar gel using
sterile borer of 6 mm diameter. The wells were filled
with 50 ìl of plant extracts of 0.1g/ml concentration
dissolved in dimethyl sulfoxide (DMSO). Ampicillin
and gentamicin (Mast dagnostics) of 10 µg per disc
were used as standard references. DMSO was used
as control. The plates were incubated at 37°C for
18-24 hours. Tests were performed in triplicate. Zone
of inhibition (ZOI) was measured in mm.
Determination of minimum inhibitory concentration
(MIC) and minimum bactericidal concentration
(MBC)
Broth dilution technique was used to determine MIC
values of the extracts which displayed antimicrobial
property following (Wiegand et al. 2008). The final
inoculum size for broth dilution was 5×105 colony-
forming units (cfu) ml-1.
Microplates were used for MIC determination. The
sterility control wells were filled with 100 µl of
MHB, and the growth control wells and wells labeled
for different concentration were filled with 50 µl of
MHB. 50 µl of stock solution of the extract (0.1g/
ml) was added and series of dilutions of the extracts
were adjusted by double dilution method. The
bacterial suspension adjusted to 1×108 cfu ml-l was
diluted to 1:100 and vortexed. Each well containing
the extract dilutions and the growth control was filled
with 50 µl of the bacterial suspension. This results
in the final desired inoculum of 5×105 cfu ml-1.
100
2019 Journal of Plant Resources Vol.17, No. 1
After incubation for 18-24 hours at 37°C, the MIC
was taken as the lowest concentration of the
antimicrobial agent that inhibited visible growth of
the tested bacteria as observed with the unaided eye.
MBC values were then determined by directly
streaking the content of the wells inhibiting bacterial
growth on MHA plates.
Results and Discussion
Magars in the study sites use root juice of F. strobilifa
in gastritis, dysentery and diarrhea, hence, the plant
material was chosen in this work. Upon successive
Soxhlet extractions of the root of F. strobilifera (100
g) using hexane and 70% methanol yielded hexane
extract (0.44g, 0.44%, light yellow) and aq.
methanolic extract (10.52 g, 10.52%, reddish black).
Phytochemical screening revealed that the hexane
extract constituted steroids, terpenoids, and the aq.
methanolic extract constituted polyphenols.
GC-MS analysis of the hexane extract led to identify
27 compounds accounting 99.37% of the total
constituents (Table 1). Out of 27 compounds, 19
hydrocarbons (60.74%), 4 fatty acids (13.18%), 1
acid ester (24.29%), 1 ester (0.55%), 1 alcohol
(0.36%) and 1 ketone (0.25%) were identified.
Octadecanoic acid was reported to be antimicrobial
(Mujeeb et al., 2014). n-Hexadecanoic acid was
reported to have antioxidant activity (Kumar et al.,
2010). (Z,Z)-9,12-Octadecadienoic acid and oleic
acid are cancer preventive and anti-inflammatory
agents (Alagammal, 2011). From the aq. methanolic
extract, 5 compounds were identified (Table 2).
Phthalic andydride (36.62%), n-hexadecanoic acid
(20.67%), 3,5-dihydroxy-6-methyl-2,3-dihydro-4H-
pyran-4-one (11.36%), 1-heptadecene (8.76%) and
octadecanoic acid (5.28%) were the main
constituents accounting 82.69% of the total
constituents. 3,5-Dihydroxy-6-methyl-2,3-dihydro-
4H-pyran-4-one was reported to be antimicrobial
agent (Kumar et al., 2010).
Magars in the study sites use root juice of F. strobilifa in gastritis, dysentery and diarrhea, hence, the
plant material was chosen in this work. Upon successive Soxhlet extractions of the root of F.
strobilifera (100 g) using hexane and 70% methanol yielded hexane extract (0.44 g, 0.44%, light
yellow) and aq. methanolic extract (10.52 g, 10.52%, reddish black). Phytochemical screening revealed
that the hexane extract constituted steroids, terpenoids, and the aq. methanolic extract constituted
polyphenols.
GC-MS analysis of the hexane extract led to identify 27 compounds accounting 99.37% of the total
constituents (Table 1). Out of 27 compounds, 19 hydrocarbons (60.74%), 4 fatty acids (13.18%), 1 acid
ester (24.29%), 1 ester (0.55%), 1 alcohol (0.36%) and 1 ketone (0.25%) were identified. Octadecanoic
acid was reported to be antimicrobial (Mujeeb et al., 2014). n-Hexadecanoic acid was reported to have
antioxidant activity (Kumar et al., 2010). (Z,Z)-9,12-Octadecadienoic acid and oleic acid are cancer
preventive and anti-inflammatory agents (Alagammal, 2011). From the aq. methanolic extract, 5
compounds were identified (Table 2). Phthalic andydride (36.62%), n-hexadecanoic acid (20.67%), 3,5-
dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one (11.36%), 1-heptadecene (8.76%) and octadecanoic
acid (5.28%) were the main constituents accounting 82.69% of the total constituents. 3,5-Dihydroxy-6-
methyl-2,3-dihydro-4H-pyran-4-one was reported to be antimicrobial agent (Kumar et al., 2010).
Table 1: Phytoconstituents identified in the hexane extract of F. strobilifera
S. N. RT Compounds Area % Nature of compound
1 26.962 4-(4-Methoxyphenyl)-2-butanone 0.25 Ketone
2 28.054 5-Phenyldecane 1.02 Hydrocarbon
3 28.299 4-Phenyldecane 0.88 Hydrocarbon
4 28.807 3-Phenyldecane 0.95 Hydrocarbon
5 29.843 2-Phenyldecane 1.43 Hydrocarbon
6 30.580 1,4a-dimethyl-7-(propan-2-ylidene)decahydronaphthalen-1-ol
(Juniper camphor)
0.36 Alcohol
7 30.760 6-Phenylundecane 2.10 Hydrocarbon
8 30.858 5-Phenylundecane 4.42 Hydrocarbon
9 31.125 4-Phenylundecane 4.54 Hydrocarbon
10 31.671 3-Phenylundecane 3.27 Hydrocarbon
11 32.675 2-Phenylundecane 4.36 Hydrocarbon
12 33.422 6-Phenyldodecane 4.93 Hydrocarbon
13 33.547 5-Phenyldodecane 4.79 Hydrocarbon
14 33.853 4-Phenyldodecane 3.61 Hydrocarbon
15 34.404 3-Phenyldodecane 3.38 Hydrocarbon
16 35.391 2-Phenyldodecane 3.98 Hydrocarbon
17 35.986 6-Phenyltridecane 5.53 Hydrocarbon
18 36.150 5-Phenyltridecane 3.40 Hydrocarbon
19 36.455 4-Phenyltridecane 2.63 Hydrocarbon
20 37.012 3-Phenyltridecane 2.70 Hydrocarbon
21 37.977 2-Phenyltridecane 2.82 Hydrocarbon
22 39.286 Butyl octyl phthalate 0.55 Ester
23 39.423 n-Hexadecanoic acid 8.32 Fatty acid
24 43.247 (Z,Z)-9,12-Octadecadienoic acid 0.68 Fatty acid
25 43.394 Oleic acid 2.01 Fatty acid
26 43.929 Octadecanoic acid 2.17 Fatty acid
27 51.670 2-(((2-ethylhexyl)oxy)carbonyl)benzoic acid 24.29 Acid ester
Table 2: Phytoconstituents identified in the methanolic extract of F. strobilifera
S. N. RT Compounds Area % Nature of compound
115.234 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one 11.36 Flavonoid fraction
221.033 Phthalic andydride 36.62 Anhydride
339.325 n-Hexadecanoic acid 20.67 Fatty acid
443.356 1-Heptadecene 8.76 Hydrocarbon
5 43.896 Octadecanoic acid 5.28 Fatty acid
100
2019 Journal of Plant Resources Vol.17, No. 1
After incubation for 18-24 hours at 37°C, the MIC
was taken as the lowest concentration of the
antimicrobial agent that inhibited visible growth of
the tested bacteria as observed with the unaided eye.
MBC values were then determined by directly
streaking the content of the wells inhibiting bacterial
growth on MHA plates.
Results and Discussion
Magars in the study sites use root juice of F. strobilifa
in gastritis, dysentery and diarrhea, hence, the plant
material was chosen in this work. Upon successive
Soxhlet extractions of the root of F. strobilifera (100
g) using hexane and 70% methanol yielded hexane
extract (0.44g, 0.44%, light yellow) and aq.
methanolic extract (10.52 g, 10.52%, reddish black).
Phytochemical screening revealed that the hexane
extract constituted steroids, terpenoids, and the aq.
methanolic extract constituted polyphenols.
GC-MS analysis of the hexane extract led to identify
27 compounds accounting 99.37% of the total
constituents (Table 1). Out of 27 compounds, 19
hydrocarbons (60.74%), 4 fatty acids (13.18%), 1
acid ester (24.29%), 1 ester (0.55%), 1 alcohol
(0.36%) and 1 ketone (0.25%) were identified.
Octadecanoic acid was reported to be antimicrobial
(Mujeeb et al., 2014). n-Hexadecanoic acid was
reported to have antioxidant activity (Kumar et al.,
2010). (Z,Z)-9,12-Octadecadienoic acid and oleic
acid are cancer preventive and anti-inflammatory
agents (Alagammal, 2011). From the aq. methanolic
extract, 5 compounds were identified (Table 2).
Phthalic andydride (36.62%), n-hexadecanoic acid
(20.67%), 3,5-dihydroxy-6-methyl-2,3-dihydro-4H-
pyran-4-one (11.36%), 1-heptadecene (8.76%) and
octadecanoic acid (5.28%) were the main
constituents accounting 82.69% of the total
constituents. 3,5-Dihydroxy-6-methyl-2,3-dihydro-
4H-pyran-4-one was reported to be antimicrobial
agent (Kumar et al., 2010).
Magars in the study sites use root juice of F. strobilifa in gastritis, dysentery and diarrhea, hence, the
plant material was chosen in this work. Upon successive Soxhlet extractions of the root of F.
strobilifera (100 g) using hexane and 70% methanol yielded hexane extract (0.44 g, 0.44%, light
yellow) and aq. methanolic extract (10.52 g, 10.52%, reddish black). Phytochemical screening revealed
that the hexane extract constituted steroids, terpenoids, and the aq. methanolic extract constituted
polyphenols.
GC-MS analysis of the hexane extract led to identify 27 compounds accounting 99.37% of the total
constituents (Table 1). Out of 27 compounds, 19 hydrocarbons (60.74%), 4 fatty acids (13.18%), 1 acid
ester (24.29%), 1 ester (0.55%), 1 alcohol (0.36%) and 1 ketone (0.25%) were identified. Octadecanoic
acid was reported to be antimicrobial (Mujeeb et al., 2014). n-Hexadecanoic acid was reported to have
antioxidant activity (Kumar et al., 2010). (Z,Z)-9,12-Octadecadienoic acid and oleic acid are cancer
preventive and anti-inflammatory agents (Alagammal, 2011). From the aq. methanolic extract, 5
compounds were identified (Table 2). Phthalic andydride (36.62%), n-hexadecanoic acid (20.67%), 3,5-
dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one (11.36%), 1-heptadecene (8.76%) and octadecanoic
acid (5.28%) were the main constituents accounting 82.69% of the total constituents. 3,5-Dihydroxy-6-
methyl-2,3-dihydro-4H-pyran-4-one was reported to be antimicrobial agent (Kumar et al., 2010).
Table 1: Phytoconstituents identified in the hexane extract of F. strobilifera
S. N. RT Compounds Area % Nature of compound
1 26.962 4-(4-Methoxyphenyl)-2-butanone 0.25 Ketone
2 28.054 5-Phenyldecane 1.02 Hydrocarbon
3 28.299 4-Phenyldecane 0.88 Hydrocarbon
4 28.807 3-Phenyldecane 0.95 Hydrocarbon
5 29.843 2-Phenyldecane 1.43 Hydrocarbon
6 30.580 1,4a-dimethyl-7-(propan-2-ylidene)decahydronaphthalen-1-ol
(Juniper camphor)
0.36 Alcohol
7 30.760 6-Phenylundecane 2.10 Hydrocarbon
8 30.858 5-Phenylundecane 4.42 Hydrocarbon
9 31.125 4-Phenylundecane 4.54 Hydrocarbon
10 31.671 3-Phenylundecane 3.27 Hydrocarbon
11 32.675 2-Phenylundecane 4.36 Hydrocarbon
12 33.422 6-Phenyldodecane 4.93 Hydrocarbon
13 33.547 5-Phenyldodecane 4.79 Hydrocarbon
14 33.853 4-Phenyldodecane 3.61 Hydrocarbon
15 34.404 3-Phenyldodecane 3.38 Hydrocarbon
16 35.391 2-Phenyldodecane 3.98 Hydrocarbon
17 35.986 6-Phenyltridecane 5.53 Hydrocarbon
18 36.150 5-Phenyltridecane 3.40 Hydrocarbon
19 36.455 4-Phenyltridecane 2.63 Hydrocarbon
20 37.012 3-Phenyltridecane 2.70 Hydrocarbon
21 37.977 2-Phenyltridecane 2.82 Hydrocarbon
22 39.286 Butyl octyl phthalate 0.55 Ester
23 39.423 n-Hexadecanoic acid 8.32 Fatty acid
24 43.247 (Z,Z)-9,12-Octadecadienoic acid 0.68 Fatty acid
25 43.394 Oleic acid 2.01 Fatty acid
26 43.929 Octadecanoic acid 2.17 Fatty acid
27 51.670 2-(((2-ethylhexyl)oxy)carbonyl)benzoic acid 24.29 Acid ester
Table 2: Phytoconstituents identified in the methanolic extract of F. strobilifera
S. N. RT Compounds Area % Nature of compound
115.234 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one 11.36 Flavonoid fraction
221.033 Phthalic andydride 36.62 Anhydride
339.325 n-Hexadecanoic acid 20.67 Fatty acid
443.356 1-Heptadecene 8.76 Hydrocarbon
5 43.896 Octadecanoic acid 5.28 Fatty acid
101
2019 Journal of Plant Resources Vol.17, No. 1
The results of antimicrobial susceptibility tests are
shown in Figures 1-3 and Table 3. The aq. methanolic
extract of the roots of F. strobilifera showed an equal
antimicrobial efficacy as gentamicin against S. typhi.
The extract exhibited potential antimicrobial activity
against E. coli (ZOI = 16.5±0.67 mm) and S. aureus
(ZOI = 15.66±0.33 mm). The extract also showed
antimicrobial activity against B. subtilis, K.
pneumoniae and P. aeruginosa which were resistant
to standard antibiotic ampicillin. S. dysenteriae,
causal bacteria of shigellosis, was also inhibited.
Madan et al. (2009) have reported antimicrobial
activity of some isoflavonoids isolated from the roots
of F. strobilifera against gram-positive (S. aureus)
and gram-negative bacteria (E. coli and P.
aeruginosa). The hexane extract showed antimicrobial
activity against S. aureus, B. subtilis, E. coli and P.
aeruginosa. The synergistic effect of various
constituents present in the extracts is responsible for
the antimicrobial activity.
Magars in the study sites use root juice of F. strobilifa in gastritis, dysentery and diarrhea, hence, the
plant material was chosen in this work. Upon successive Soxhlet extractions of the root of F.
strobilifera (100 g) using hexane and 70% methanol yielded hexane extract (0.44 g, 0.44%, light
yellow) and aq. methanolic extract (10.52 g, 10.52%, reddish black). Phytochemical screening revealed
that the hexane extract constituted steroids, terpenoids, and the aq. methanolic extract constituted
polyphenols.
GC-MS analysis of the hexane extract led to identify 27 compounds accounting 99.37% of the total
constituents (Table 1). Out of 27 compounds, 19 hydrocarbons (60.74%), 4 fatty acids (13.18%), 1 acid
ester (24.29%), 1 ester (0.55%), 1 alcohol (0.36%) and 1 ketone (0.25%) were identified. Octadecanoic
acid was reported to be antimicrobial (Mujeeb et al., 2014). n-Hexadecanoic acid was reported to have
antioxidant activity (Kumar et al., 2010). (Z,Z)-9,12-Octadecadienoic acid and oleic acid are cancer
preventive and anti-inflammatory agents (Alagammal, 2011). From the aq. methanolic extract, 5
compounds were identified (Table 2). Phthalic andydride (36.62%), n-hexadecanoic acid (20.67%), 3,5-
dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one (11.36%), 1-heptadecene (8.76%) and octadecanoic
acid (5.28%) were the main constituents accounting 82.69% of the total constituents. 3,5-Dihydroxy-6-
methyl-2,3-dihydro-4H-pyran-4-one was reported to be antimicrobial agent (Kumar et al., 2010).
Table 1: Phytoconstituents identified in the hexane extract of F. strobilifera
S. N. RT Compounds Area % Nature of compound
1 26.962 4-(4-Methoxyphenyl)-2-butanone 0.25 Ketone
2 28.054 5-Phenyldecane 1.02 Hydrocarbon
3 28.299 4-Phenyldecane 0.88 Hydrocarbon
4 28.807 3-Phenyldecane 0.95 Hydrocarbon
5 29.843 2-Phenyldecane 1.43 Hydrocarbon
6 30.580 1,4a-dimethyl-7-(propan-2-ylidene)decahydronaphthalen-1-ol
(Juniper camphor)
0.36 Alcohol
7 30.760 6-Phenylundecane 2.10 Hydrocarbon
8 30.858 5-Phenylundecane 4.42 Hydrocarbon
9 31.125 4-Phenylundecane 4.54 Hydrocarbon
10 31.671 3-Phenylundecane 3.27 Hydrocarbon
11 32.675 2-Phenylundecane 4.36 Hydrocarbon
12 33.422 6-Phenyldodecane 4.93 Hydrocarbon
13 33.547 5-Phenyldodecane 4.79 Hydrocarbon
14 33.853 4-Phenyldodecane 3.61 Hydrocarbon
15 34.404 3-Phenyldodecane 3.38 Hydrocarbon
16 35.391 2-Phenyldodecane 3.98 Hydrocarbon
17 35.986 6-Phenyltridecane 5.53 Hydrocarbon
18 36.150 5-Phenyltridecane 3.40 Hydrocarbon
19 36.455 4-Phenyltridecane 2.63 Hydrocarbon
20 37.012 3-Phenyltridecane 2.70 Hydrocarbon
21 37.977 2-Phenyltridecane 2.82 Hydrocarbon
22 39.286 Butyl octyl phthalate 0.55 Ester
23 39.423 n-Hexadecanoic acid 8.32 Fatty acid
24 43.247 (Z,Z)-9,12-Octadecadienoic acid 0.68 Fatty acid
25 43.394 Oleic acid 2.01 Fatty acid
26 43.929 Octadecanoic acid 2.17 Fatty acid
27 51.670 2-(((2-ethylhexyl)oxy)carbonyl)benzoic acid 24.29 Acid ester
Table 2: Phytoconstituents identified in the methanolic extract of F. strobilifera
S. N. RT Compounds Area % Nature of compound
1 15.234 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one 11.36 Flavonoid fraction
221.033 Phthalic andydride 36.62 Anhydride
339.325 n-Hexadecanoic acid 20.67 Fatty acid
443.356 1-Heptadecene 8.76 Hydrocarbon
5 43.896 Octadecanoic acid 5.28 Fatty acid
The results of antimicrobial susceptibility tests are shown in Figures 1-3 and Table 3. The aq.
methanolic extract of the roots of F. strobilifera showed an equal antimicrobial efficacy as gentamicin
against S. typhi. The extract exhibited potential antimicrobial activity against E. coli (ZOI =
16.5±0.67 mm) and S. aureus (ZOI = 15.66±0.33 mm). The extract also showed antimicrobial activity
against B. subtilis,K. pneumoniae and P. aeruginosa which were resistant to standard antibiotic
ampicillin. S. dysenteriae, causal bacteria of shigellosis, was also inhibited. Madan et al. (2009) have
reported antimicrobial activity of some isoflavonoids isolated from the roots of F. strobilifera against
gram-positive (S. aureus) and gram-negative bacteria (E. coli and P. aeruginosa). The hexane extract
showed antimicrobial activity against S. aureus, B. subtilis, E. coli and P. aeruginosa. The synergistic
effect of various constituents present in the extracts is responsible for the antimicrobial activity.
Figure 1: The aq. methanolic extract
showing antibacterial activity
against B. subtilis. Ampicillin as +ve
control.
Figure 2: The aq. methanolic
extract showing antibacterial activity
against S. typhi. Ampicillin as +ve
control.
Figure 3: The aq. methanolic
extract showing antibacterial
activity against S. aureus.
Ampicillin as +ve control.
Table 3: Antimicrobial activity of F. strobilifera
S.
N
Sample Diameter of inhibition zone (mm)±standard error mean (SEM)
Gram positive bacteria Gram negative bacteria
SA BS EF EC ST KP PA SD
1 HE 11.25±0.47 13.3±0.33 - 11.66±0.33 - - 12.33±0.66 -
2 ME 15.66±0.33 13.33±0.88 11.66±0.33 16.5±0.67 12.66±0.33 11.66±0.33 11.8±0.33 10.33±0.33
3 Ampicillin 32.5±0.5 8.5±0.5 17.75±0.25 25±1 15.5±0.5 8.5±0.5 - 23.75±0.25
4 Gentamicin 16.75±0.25 15.5±0.5 18.5±0.5 17.5±0.5 12.66±0.33 11.33±0.88 14.66±0.33 18.66±0.66
5 DMSO - - - - - - - -
HE = Hexane extract, ME = Aq. methanolic extract, SA = S. aureus,BS = B. subtilis,EF = E. faecalis,EC = E. coli,ST =
S. typhi,KP = K. pneumoniae,PA = P. aeruginosa,SD = S. dysenteriae
MIC and MBC values are shown in Table 4. Lowest MIC was found to be 1.56 mg/ml for the aq.
methnolic extract against E. coli. The extract showed bactericidal effect on B. subtilis, E. faecalis, K.
pneumoniae and S. dysenteriae. It showed bactericidal effect on higher concentration against S. aureus,
E. coli, S. typhi and P. aeruginosa. The bacterial viability was gradually decreased at high concentration
of the extract in a dose-dependent manner.
Table 4: MIC and MBC of F. strobilifera
S. N. Bacteria Hexane extract Aq. methanolic extract
MIC MBC MIC MBC
1S. aureus 12.5 25 3.12 6.25
2B. subtilis 50 50 12.5 12.5
3E. faecalis - - 3.12 3.12
4E. coli 6.25 6.25 1.56 6.25
5S. typhi - - 12.5 25
6K. pneumoniae - - 6.25 6.25
7P. aeruginosa 12.5 25 3.12 6.25
8S. dysenteriae - - 50 50
The results of antimicrobial susceptibility tests are shown in Figures 1-3 and Table 3. The aq.
methanolic extract of the roots of F. strobilifera showed an equal antimicrobial efficacy as gentamicin
against S. typhi. The extract exhibited potential antimicrobial activity against E. coli (ZOI =
16.5±0.67 mm) and S. aureus (ZOI = 15.66±0.33 mm). The extract also showed antimicrobial activity
against B. subtilis,K. pneumoniae and P. aeruginosa which were resistant to standard antibiotic
ampicillin. S. dysenteriae, causal bacteria of shigellosis, was also inhibited. Madan et al. (2009) have
reported antimicrobial activity of some isoflavonoids isolated from the roots of F. strobilifera against
gram-positive (S. aureus) and gram-negative bacteria (E. coli and P. aeruginosa). The hexane extract
showed antimicrobial activity against S. aureus, B. subtilis, E. coli and P. aeruginosa. The synergistic
effect of various constituents present in the extracts is responsible for the antimicrobial activity.
Figure 1: The aq. methanolic extract
showing antibacterial activity
against B. subtilis. Ampicillin as +ve
control.
Figure 2: The aq. methanolic
extract showing antibacterial activity
against S. typhi. Ampicillin as +ve
control.
Figure 3: The aq. methanolic
extract showing antibacterial
activity against S. aureus.
Ampicillin as +ve control.
Table 3: Antimicrobial activity of F. strobilifera
S. Sample Diameter of inhibition zone (mm)±standard error mean (SEM)
Gram positive bacteria Gram negative bacteria
SA BS EF EC ST KP PA SD
1 HE 11.25±0.47 13.3±0.33 - 11.66±0.33 - - 12.33±0.66 -
2 ME 15.66±0.33 13.33±0.88 11.66±0.33 16.5±0.67 12.66±0.33 11.66±0.33 11.8±0.33 10.33±0.33
3 Ampicillin 32.5±0.5 8.5±0.5 17.75±0.25 25±1 15.5±0.5 8.5±0.5 - 23.75±0.25
4 Gentamicin 16.75±0.25 15.5±0.5 18.5±0.5 17.5±0.5 12.66±0.33 11.33±0.88 14.66±0.33 18.66±0.66
5 DMSO - - - - - - - -
HE = Hexane extract, ME = Aq. methanolic extract, SA = S. aureus,BS = B. subtilis,EF = E. faecalis,EC = E. coli,ST =
S. typhi,KP = K. pneumoniae,PA = P. aeruginosa,SD = S. dysenteriae
MIC and MBC values are shown in Table 4. Lowest MIC was found to be 1.56 mg/ml for the aq.
methnolic extract against E. coli. The extract showed bactericidal effect on B. subtilis, E. faecalis, K.
pneumoniae and S. dysenteriae. It showed bactericidal effect on higher concentration against S. aureus,
E. coli, S. typhi and P. aeruginosa. The bacterial viability was gradually decreased at high concentration
of the extract in a dose-dependent manner.
Table 4: MIC and MBC of F. strobilifera
S. N. Bacteria Hexane extract Aq. methanolic extract
MIC MBC MIC MBC
1S. aureus 12.5 25 3.12 6.25
2B. subtilis 50 50 12.5 12.5
3E. faecalis - - 3.12 3.12
4E. coli 6.25 6.25 1.56 6.25
5S. typhi - - 12.5 25
6K. pneumoniae - - 6.25 6.25
7P. aeruginosa 12.5 25 3.12 6.25
8S. dysenteriae - - 50 50
Figure 1: The aq. methanolic extract
showing antibacterial activity against B.
subtilis. Ampicillin as +ve control.
Figure 2: The aq. methanolic extract
showing antibacterial activity against S.
typhi. Ampicillin as +ve control.
Figure 3: The aq. methanolic extract
showing antibacterial activity against S.
aureus. Ampicillin as +ve control.
MIC and MBC values are shown in Table 4. Lowest
MIC was found to be 1.56 mg/ml for the aq.
methnolic extract against E. coli. The extract showed
bactericidal effect on B. subtilis, E. faecalis, K.
pneumoniae and S. dysenteriae. It showed
bactericidal effect on higher concentration against
S. aureus, E. coli, S. typhi and P. aeruginosa. The
bacterial viability was gradually decreased at high
concentration of the extract in a dose-dependent
manner.
102
2019 Journal of Plant Resources Vol.17, No. 1
Conclusion
The people of Magar communities of Kawaswoti
rural municipality, Nawalpur district, Province no.
4, Nepal use juice from the roots of F. strobilifera
for the treatment of gastritis, diarrhea and dysentery.
This work showed that aq. methanolic extract of F.
strobilifera roots exhihbit significant antimicrobial
activity against E. coli (ZOI = 16.5±0.67) and
moderately against S. dysenteriae (ZOI =
10.33±0.33) in the support of traditional knowledge.
The extract also displayed antimicrobial activity
against ampicillin-resistant B. subtilis, K.
pneumoniae and P. aeruginosa.
Acknowledgements
We are grateful to Tri-Chandra Multiple Campus for
forwarding necessary administrative efforts for this
research study. We would like to express our thanks
to the University Grant Commission (UGC) for
providing grant for the research. We are grateful to
Nepal Academy of Science and Technology (NAST)
for providing necessary laboratory facilities for the
research. Our special thanks go to local villagers,
healers and informants for their cooperation. We are
deeply indebted to the National Herbarium and Plant
Laboratories, Godavari, Lalitpur for plant
identification.
References
Alagammal, M., Tresina, S.P., & Mohan, V.R.
(2011). Chemical investigations of Polygala
chinensis L. by GC-MS. Science Research
Reporter, 1(2), 4952.
Bhattarai, N.K. (1991). Folk herbal medicines of
Makawanpur district, Nepal. International
Journal of Pharmacognosy, 29(4), 284295.
Ciulei, I. (1982). Methods for studying vegetables
drugs. Bucharest, Romania: Chemical Industries
Branch, Division of Industrial Operations,
UNIDO.
Dangol, D.R., & Gurung, S.B. (2000). Ethno-
botanical study of Darai tribe of Chitwan district,
Nepal. Proceeding of the Third National
Conference on Science and Technology, vol 2 (pp.
11941213). Royal Nepal Academy of Science
and Technology, Kathmandu, Nepal.
Ghalot, K., Lal, V.K., & Jha, S. (2011).
Phytochemical and pharmacological potential of
Flemingia Roxb. ex W.T. Aiton (Fabaceae).
International Journal of Phytomedicine, 3, 294
307.
Kumar, A., Dora, J., Gahlot, K., & Tripathi, R.
(2011a). Anthelmintic activity of Flemingia
strobilifera ( R. Br ). International Journal of
Research in Pharmaceutical and Biomedical
Sciences, 2(3), 10771078.
Kumar, A., Gahlot, K., Dora, J., & Singh, P. (2011b).
Analgesic activity of methanolic extract of
Flemingia strobilifera (R. Br). International
Journal of Research in Pharmacy and Chemistry,
1(4), 825827.
Kumar, P.P., Kumaravel S., & Lalitha, C. (2010).
Screening of antioxidant activity, total phenolics
and GC-MS study of Vitex negundo. African
Journal of Biochemistry Research, 4(7), 191195.
The results of antimicrobial susceptibility tests are shown in Figures 1-3 and Table 3. The aq.
methanolic extract of the roots of F. strobilifera showed an equal antimicrobial efficacy as gentamicin
against S. typhi. The extract exhibited potential antimicrobial activity against E. coli (ZOI =
16.5±0.67 mm) and S. aureus (ZOI = 15.66±0.33 mm). The extract also showed antimicrobial activity
against B. subtilis,K. pneumoniae and P. aeruginosa which were resistant to standard antibiotic
ampicillin. S. dysenteriae, causal bacteria of shigellosis, was also inhibited. Madan et al. (2009) have
reported antimicrobial activity of some isoflavonoids isolated from the roots of F. strobilifera against
gram-positive (S. aureus) and gram-negative bacteria (E. coli and P. aeruginosa). The hexane extract
showed antimicrobial activity against S. aureus, B. subtilis, E. coli and P. aeruginosa. The synergistic
effect of various constituents present in the extracts is responsible for the antimicrobial activity.
Figure 1: The aq. methanolic extract
showing antibacterial activity
against B. subtilis. Ampicillin as +ve
control.
Figure 2: The aq. methanolic
extract showing antibacterial activity
against S. typhi. Ampicillin as +ve
control.
Figure 3: The aq. methanolic
extract showing antibacterial
activity against S. aureus.
Ampicillin as +ve control.
Table 3: Antimicrobial activity of F. strobilifera
S. Sample Diameter of inhibition zone (mm)±standard error mean (SEM)
Gram positive bacteria Gram negative bacteria
SA BS EF EC ST KP PA SD
1 HE 11.25±0.47 13.3±0.33 - 11.66±0.33 - - 12.33±0.66 -
2 ME 15.66±0.33 13.33±0.88 11.66±0.33 16.5±0.67 12.66±0.33 11.66±0.33 11.8±0.33 10.33±0.33
3 Ampicillin 32.5±0.5 8.5±0.5 17.75±0.25 25±1 15.5±0.5 8.5±0.5 - 23.75±0.25
4 Gentamicin 16.75±0.25 15.5±0.5 18.5±0.5 17.5±0.5 12.66±0.33 11.33±0.88 14.66±0.33 18.66±0.66
5 DMSO - - - - - - - -
HE = Hexane extract, ME = Aq. methanolic extract, SA = S. aureus,BS = B. subtilis,EF = E. faecalis,EC = E. coli,ST =
S. typhi,KP = K. pneumoniae,PA = P. aeruginosa,SD = S. dysenteriae
MIC and MBC values are shown in Table 4. Lowest MIC was found to be 1.56 mg/ml for the aq.
methnolic extract against E. coli. The extract showed bactericidal effect on B. subtilis, E. faecalis, K.
pneumoniae and S. dysenteriae. It showed bactericidal effect on higher concentration against S. aureus,
E. coli, S. typhi and P. aeruginosa. The bacterial viability was gradually decreased at high concentration
of the extract in a dose-dependent manner.
Table 4: MIC and MBC of F. strobilifera
S. N. Bacteria Hexane extract Aq. methanolic extract
MIC MBC MIC MBC
1S. aureus 12.5 25 3.12 6.25
2B. subtilis 50 50 12.5 12.5
3E. faecalis - - 3.12 3.12
4E. coli 6.25 6.25 1.56 6.25
5S. typhi - - 12.5 25
6K. pneumoniae - - 6.25 6.25
7P. aeruginosa 12.5 25 3.12 6.25
8S. dysenteriae - - 50 50
102
2019 Journal of Plant Resources Vol.17, No. 1
Conclusion
The people of Magar communities of Kawaswoti
rural municipality, Nawalpur district, Province no.
4, Nepal use juice from the roots of F. strobilifera
for the treatment of gastritis, diarrhea and dysentery.
This work showed that aq. methanolic extract of F.
strobilifera roots exhihbit significant antimicrobial
activity against E. coli (ZOI = 16.5±0.67) and
moderately against S. dysenteriae (ZOI =
10.33±0.33) in the support of traditional knowledge.
The extract also displayed antimicrobial activity
against ampicillin-resistant B. subtilis, K.
pneumoniae and P. aeruginosa.
Acknowledgements
We are grateful to Tri-Chandra Multiple Campus for
forwarding necessary administrative efforts for this
research study. We would like to express our thanks
to the University Grant Commission (UGC) for
providing grant for the research. We are grateful to
Nepal Academy of Science and Technology (NAST)
for providing necessary laboratory facilities for the
research. Our special thanks go to local villagers,
healers and informants for their cooperation. We are
deeply indebted to the National Herbarium and Plant
Laboratories, Godavari, Lalitpur for plant
identification.
References
Alagammal, M., Tresina, S.P., & Mohan, V.R.
(2011). Chemical investigations of Polygala
chinensis L. by GC-MS. Science Research
Reporter, 1(2), 4952.
Bhattarai, N.K. (1991). Folk herbal medicines of
Makawanpur district, Nepal. International
Journal of Pharmacognosy, 29(4), 284295.
Ciulei, I. (1982). Methods for studying vegetables
drugs. Bucharest, Romania: Chemical Industries
Branch, Division of Industrial Operations,
UNIDO.
Dangol, D.R., & Gurung, S.B. (2000). Ethno-
botanical study of Darai tribe of Chitwan district,
Nepal. Proceeding of the Third National
Conference on Science and Technology, vol 2 (pp.
11941213). Royal Nepal Academy of Science
and Technology, Kathmandu, Nepal.
Ghalot, K., Lal, V.K., & Jha, S. (2011).
Phytochemical and pharmacological potential of
Flemingia Roxb. ex W.T. Aiton (Fabaceae).
International Journal of Phytomedicine, 3, 294
307.
Kumar, A., Dora, J., Gahlot, K., & Tripathi, R.
(2011a). Anthelmintic activity of Flemingia
strobilifera ( R. Br ). International Journal of
Research in Pharmaceutical and Biomedical
Sciences, 2(3), 10771078.
Kumar, A., Gahlot, K., Dora, J., & Singh, P. (2011b).
Analgesic activity of methanolic extract of
Flemingia strobilifera (R. Br). International
Journal of Research in Pharmacy and Chemistry,
1(4), 825827.
Kumar, P.P., Kumaravel S., & Lalitha, C. (2010).
Screening of antioxidant activity, total phenolics
and GC-MS study of Vitex negundo. African
Journal of Biochemistry Research, 4(7), 191195.
The results of antimicrobial susceptibility tests are shown in Figures 1-3 and Table 3. The aq.
methanolic extract of the roots of F. strobilifera showed an equal antimicrobial efficacy as gentamicin
against S. typhi. The extract exhibited potential antimicrobial activity against E. coli (ZOI =
16.5±0.67 mm) and S. aureus (ZOI = 15.66±0.33 mm). The extract also showed antimicrobial activity
against B. subtilis,K. pneumoniae and P. aeruginosa which were resistant to standard antibiotic
ampicillin. S. dysenteriae, causal bacteria of shigellosis, was also inhibited. Madan et al. (2009) have
reported antimicrobial activity of some isoflavonoids isolated from the roots of F. strobilifera against
gram-positive (S. aureus) and gram-negative bacteria (E. coli and P. aeruginosa). The hexane extract
showed antimicrobial activity against S. aureus, B. subtilis, E. coli and P. aeruginosa. The synergistic
effect of various constituents present in the extracts is responsible for the antimicrobial activity.
Figure 1: The aq. methanolic extract
showing antibacterial activity
against B. subtilis. Ampicillin as +ve
control.
Figure 2: The aq. methanolic
extract showing antibacterial activity
against S. typhi. Ampicillin as +ve
control.
Figure 3: The aq. methanolic
extract showing antibacterial
activity against S. aureus.
Ampicillin as +ve control.
Table 3: Antimicrobial activity of F. strobilifera
S. Sample Diameter of inhibition zone (mm)±standard error mean (SEM)
Gram positive bacteria Gram negative bacteria
SA BS EF EC ST KP PA SD
1 HE 11.25±0.47 13.3±0.33 - 11.66±0.33 - - 12.33±0.66 -
2 ME 15.66±0.33 13.33±0.88 11.66±0.33 16.5±0.67 12.66±0.33 11.66±0.33 11.8±0.33 10.33±0.33
3 Ampicillin 32.5±0.5 8.5±0.5 17.75±0.25 25±1 15.5±0.5 8.5±0.5 - 23.75±0.25
4 Gentamicin 16.75±0.25 15.5±0.5 18.5±0.5 17.5±0.5 12.66±0.33 11.33±0.88 14.66±0.33 18.66±0.66
5 DMSO - - - - - - - -
HE = Hexane extract, ME = Aq. methanolic extract, SA = S. aureus,BS = B. subtilis,EF = E. faecalis,EC = E. coli,ST =
S. typhi,KP = K. pneumoniae,PA = P. aeruginosa,SD = S. dysenteriae
MIC and MBC values are shown in Table 4. Lowest MIC was found to be 1.56 mg/ml for the aq.
methnolic extract against E. coli. The extract showed bactericidal effect on B. subtilis, E. faecalis, K.
pneumoniae and S. dysenteriae. It showed bactericidal effect on higher concentration against S. aureus,
E. coli, S. typhi and P. aeruginosa. The bacterial viability was gradually decreased at high concentration
of the extract in a dose-dependent manner.
Table 4: MIC and MBC of F. strobilifera
S. N. Bacteria Hexane extract Aq. methanolic extract
MIC MBC MIC MBC
1S. aureus 12.5 25 3.12 6.25
2B. subtilis 50 50 12.5 12.5
3E. faecalis - - 3.12 3.12
4E. coli 6.25 6.25 1.56 6.25
5S. typhi - - 12.5 25
6K. pneumoniae - - 6.25 6.25
7P. aeruginosa 12.5 25 3.12 6.25
8S. dysenteriae - - 50 50
103
2019 Journal of Plant Resources Vol.17, No. 1
Madan, S., Singh, G.N., Kumar, Y., & Kohli, K.
(2010). Phytochemical analysis and free-radical
scavenging activity of Flemingia strobilifera
(Linn) R. Br. Research Journal of
Pharmaceutical, Biological and Chemical
Sciences, 1(4), 183190.
Madan, S., Singh, G.N., Kohli, K., Ali, M., Kumar,
Y., Singh, R.M., & Prakash, O. (2009).
Isoflavonoids from Flemingia strobilifera (L) R.
Br. roots. Acta Poloniae Pharmaceutica-Drug
Research, 66(3), 297303.
Manandhar, N. P. (2002). Plants and people of
Nepal. Portland Oregon, USA: Timber Press.
Mujeeb, F., Bajpai, P., & Pathak, N. (2014).
Phytochemical evaluation, antimicrobial activity,
and determination of bioactive components from
leaves of Aegle marmelos. BioMed Research
International. http://dx.doi.org/10.1155/2014/
497606.
Nemkul, C.M., Bajracharya, G.B., & Shrestha, I.
(2018). Phytochemical, antibacterial and DPPH
free radical scavenging evaluations of the barks
of Aegle marmelos (L.) Correa. Journal of
Pharmacognosy and Phytochemistry, 7(4), 1637
1641.
Perez, C., Pauli, M. & Bazerque, P. (1990). An
antibiotic assay by the agar-well diffusion
method. Acta Biologiae et Medecine
Experimentalis, 15, 113115.
Rijal, A. (2011). Surviving on knowledge:
ethnobotany of Chepang community from mid-
hills of Nepal. A Journal of Plants, People and
Applied Research, 9, 181215.
Wiegand, I., Hilpert, K., & Hancock, R.E.W. (2008).
Agar and broth dilution methods to determine the
minimal inhibitory concentration (MIC) of
antimicrobial substances. Nature Protocols, 3(2),
163175.