Gangai Abirami and Nirmala., (2013) Int. J. Res. Phytochem. Pharmacol.

, 3(2), 103-105
JK Welfare &Pharmascope Foundation | International Journal of Research in Phytochemistry& Pharmacology 103





A comparative phytochemical and anti-bacterial study of Menthapiperita and
Citrus sinensis
Gangai Abirami S. K
1
, Nirmala P
2

1
Department of Biotechnology, Kongu Arts and Science College, Erode, India
2
Department of Biotechnology, Nehru arts and Science College, Coimbatore, India
ABSTRACT
An in vitro study was carried out to screen the antibacterial activity of four different extracts of Menthapiperita
and Citrus sinensis against six pathogenic bacteria Bacillus sublilis, Staphylococcus aureus, Escherichia coli, Salmo-
nella typhimurium, Proteus vulgaris, and Klebsiellapneumoniae. Highest anti bacterial activity was observed in ace-
tone extract of Menthapiperita leaves followed by aqueous extracts of Citrus sinensis fruit peel. Preliminary phy-
tochemical screening of the plants shows the presence of alkaloids, flavanoids and tannins in Menthapiperita and
alkaloids, flavanoids, terpenoids and tannins in Citrus sinensis.
Keywords: Antibacterial activity; phytochemicals; acetone; aqueous extract
INTRODUCTION
Plants are the local heritage with global importance.
World is endowed with a rich wealth of medicinal
plants. The knowledge of the chemical constituents of
plants wouldfurther be valuable in discovering the ac-
tual value of folk remedies.In recent year, essential oil
and herbal extracts have attracted a great scientific
interest due to their potential use as a source of natu-
ral antioxidants and biologically active compounds
(Wannissom et al., 2005). There is a growing tendency
all over the world, to shift from synthetic to natural
based products including medicinal and aromatic
plants.
Man has exploited mint species for more than 2000
years. The genus Mentha (Lamiaceae)iscomposedof19
geographically widespread species with13hybrids (Ra-
verchon, 2006). Menthapiperita (Pepper mint) has a
long tradition of medicinal use, with archaeological
evidence placing its use at least as far back as ten thou-
sand years. Peppermint has high menthol content, and
is often used in tea and for flavouring ice cream, con-
fectionery, chewing gum and toothpaste. The oil also
contains menthone and menthyl esters, particularly
menthyl acetate. (PDR for Herbal Medicines)
Citrus is one of the most important commercial food
crops grown in all continents of the world (Tao et al.,
2007). Citrus fruits are mainly used by juice processing
industries while the peels are generally wasted. There
is always an increased attention in bringinguseful
products from waste materials and citrus wastes are no
exceptions. Epidermal content of citrus fruit is rich in
flavanoids and many poly methoxylated flavones
(Ahmed et al., 2006). The anti microbial abilities of es-
sential oil, among which citrus oils are also shown to be
a particularly interesting field for applications in the
food and cosmetic industries (Caccioni et al., 1998).In
view of the vast potentiality of aromatic plants, the
present work was undertaken to compare the two
aromatic plants and evaluate the presence of phyto-
chemical constituents and their antibacterial activities.
MATERIALSAND METHODS
The samples were collected from Erode district. They
are anatomically identified as Menthapiperita and Ci-
trus sinensis and authenticated by Dr. M. Palanisamy,
Botanical Survey of India, TNAU - Coimbatore. Epider-
mal content of fruit, leaves and stems of Menthapiperi-
tawere collected, washed and dried in shade for a pe-
riod of 5 days at room temperature (32-35C). The
dried plant parts were finely powdered and reduced.
The powder was transferred in to closed containers.
Preparation of Extracts
Powdered plant materials were separately extracted
with different solvents (Methanol, Ethyl acetate, ace-
tone, and Water) for 15hrs.The extracted materials
were dried at 60C in vaccum evaporator for
10hrs.Dried extracts were dissolved in sterile DMSO at
a concentration of 20mg, 40mg, 60mg, 80mg and
100mg and the extracts were refrigerated.
Phytochemical Analysis
Phytochemical analysis was carried out as described
(Venkata et al., 2010).

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ISSN: 2231-010X
Research Article

* Corresponding Author
Email: mithrajabi@gmail.com
Received on: 12-03-2013
Revised on: 22-04-2013
Accepted on: 23-04-2103

Gangai Abirami and Nirmala., (2013) Int. J. Res. Phytochem. Pharmacol., 3(2), 103-105
104 JK Welfare &Pharmascope Foundation | International Journal of Research in Phytochemistry& Pharmacology

In-vitro testing of extracts for antibacterial activity
Micro Organisms
Clinically isolated pathogenic bacteria were confirmed
with biochemical assays. Staphylococcus aureus, Bacil-
lus subtilis, Escherichia coli, Proteus vulgaris, Salmonel-
la typhimurium and Klebsiellapuemoniaewere grown in
Nutrient broth and stored at 4C. Active log phase
growing microorganisms were used for antibacterial
activity.
Antibacterial activity by agar diffusion method
Agar well diffusion method (Sabahatsaeed et al., 2005)
was used to test the antibacterial activity against the
pathogenic bacteria. Muller Hinton agar plates were
spreaded with 50 l of log phase. Bacterial culture wells
were made in the agar plate with sterile cork borer
(8mm diameter). 50% of each extract, at different con-
centration were introduced in the well and allowed to
diffuse. The plates were incubated at 37C for 24 hours
and zone of inhibition was measured in millimeter
(mm).
Statistical Analysis
Results were analysed with student t test and Analysis
of Variance (ANOVA). Mean value were expressed by
standard deviation SD+ 0.01
RESULT
The preliminary Phytochemical screening for leaves,
(leaf and stem) of Menthapiperita shown the presence
of tannins and alkaloids in ethyl acetate and methanol
extract.
Aqueous, Methanol, Ethyl acetate and Acetone ex-
tracts of Citrus sinensis contain flavonoids. Tannins
were found in acetone and aqueous extracts. Terpe-
noidswere found in acetone, ethyl acetate and metha-
nol extracts. (Table-1).S.aureus was found to be highly
sensitive (24mm) against acetone extract of Menthapi-
perita. B.subtilis was moderately sensitive (22mm)
against acetone and aqueous extracts of Menthapiperi-
ta.(Table-2).
Among the gram-negative bacteria tested, S.typhi
(20mm) was found to be the most susceptible to ace-
tone extract of Menthapiperita as compared with
P.vulgaris (18mm) and E.coli (18mm) in acetone and
ethyl acetate extracts.
S.typhi was found to be highly sensitive (17mm) against
aqueous and acetone extract of C. Sinensis, P.vulgaris
(16mm) and E.coli (15mm)was to be sensitive against
aqueous and acetone extract respectively.
DISCUSSION
The study showed that acetone extract of Menthapipe-
rita has potent antibacterial activity against S. aureus.
The leaf and leaf+ stem are rich source of secondary
phytoconstituents, which impart their therapeutic ef-
fects and use of this herb in folk medicines.
This work has also identified the aqueous extract of
Citrus sinensis had its vast potency against S. aureus
and also the secondary metabolites in peels obtained
from different solvent extracts. This shows that, the
Table 1: Phytochemical screening for Menthaarvensisand Citrus sinensis
Phytochemical
Methanol Ethyl acetate Acetone Aqueous
M. pi-
perita
C.
sinensis
M. pi-
perita
C.sinensis
M. pi-
perita
C.sinensis
M. pi-
perita
C.sinensis
L L+s E L L+s E L L+s E L L+s E
Alkaloids - - + + + + + + - - - -
Flavonoids + + + - - - - - + - - +
Phenols - - - - - - - - - - - -
Terpenoids - - + - - - - - + - - +
Tannins + + + - - - - - + - - +
L Leaf., S Stem., E Epidermal layer
Table 2: Antibacterial activity for Menthaarvensis and Citrus sinensis
Microorganism
Methanol Ethyl acetate Acetone Aqueous
M. pi-
perita
C.
sinensis
M. pi-
perita
C.
sinensis
M. pi-
perita
C.
sinensis
M. pi-
perita
C.
sinensis
B. subtilis 21 20 12 20 22 16 17 14
S. aureus 7 - 12 12 24 21 22 21
S. typhi 11 19 17 12 20 17 16 17
P. vulgaris - 15 14 14 18 10 13 16
E. coli 6 11 18 15 16 11 13 11
K. pneumoniae - - - - - - - -

Gangai Abirami and Nirmala., (2013) Int. J. Res. Phytochem. Pharmacol., 3(2), 103-105
JK Welfare &Pharmascope Foundation | International Journal of Research in Phytochemistry& Pharmacology 105

peel of sweet orange is not only an astringent but also
is a good antibacterial agent.
Klebsiella pneumonia does not show any activity. The
negative results do not mean the absence of bioactive
constituents nor is that the plant inactive. Active com-
pounds may be present in insufficient quantities in the
crude extracts to show activity with the dose levels
employed (Taylor et al., 2011). Using large doses can
thus prove lack of activity.

Figure 1: Antibacterial activity of Mentaarvensis

Figure 2: Antibacterial activity of Citrus sinensis
CONCLUSION
Further research is necessary to find the bioactive
compounds contributing to the antibacterial activity
and accelerating the transformation by putting science
into the art of traditional medicine.
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