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HPLC Analysis of Methanolic Extract of Some Medicinal Plant Leaves of

Myrtaceae Family

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 INTERNATIONALE PHARMACEUTICA SCIENCIA
| July September 2012 | Vol. 2 | Issue 3 |
Available online http://www.ipharmsciencia.com
ISSN 2231-5896
©2012 IPS
RESEARCH ARTICLE

HPLC Analysis of Methanolic Extract of Some Medicinal Plant Leaves of Myrtaceae

Family
ABSTRACT Tripathi, I.P.a, Mishra, Mahendra
KR.*b, Pardhi, Yogeshc, Dwivedi, Atulb,
In the present study, we have applied easiest and quicker method for Dwivedi, Noopab, Kamal, Artib, Gupta,
extraction of methanol soluble phytochemicals. Crude extract of plant Priyankab.
leaves (Psidium guajava, Syzygium cumini, Eucalypts) was analyzed the aFaculty of Science and Environment,
soluble fraction of phenols using HPLC coupled to UV-Vis detector at 254 Mahatma Gandhi Chitrakoot Gramoday
nm. Which gives different peaks at this region, it may be phenolic Vishwavidyalaya.
bResearch Scholar, Mahatma Gandhi
compounds.
Chitrakoot Gramoday Vishwavidyalaya.
c Research Associate, Tropical forest

Research Institute, Jabalpur.

*Mahendra Kr. Mishra (Research Scholar)
Dept. of Physical Science,
Keywords: Psidium guajava, Syzygium cumini, Eucalypts, Mahatma Gandhi Chitrakoot Gramoday
Vishwavidyalaya, Chitrakoot (M.P)
Phytochemicals, Phenols, Extraction, HPLC.
Date of Submission: 12-07-2012
Date of Acceptance: 02-08-2012
Conflict of interest: Nil
Source of support: None

Introduction a dicot family in the class Rosidae, which also includes

Phytochemicals are naturally occurring,
biologically active chemical compounds in plants. The
prefix “Phyto” is from a Greek word meaning plant. In
plants, phytochemicals act as a natural defense system
for host plants and provide colour, aroma and flavour.
More than 4000 of these compounds have been
discovered to date and it is expected that scientists will
discover many more. Historically, plants have provided
a source of inspiration for novel drug compounds, as
plant derived medicines have made large contributions
to human health and well being. Their role is twofold
in the development of new drugs[1]. Natural products
and secondary metabolites formed by living systems,
notably from plant origin, have shown great potential
in treating human diseases such as cancer, coronary
heart diseases, diabetes and infectious diseases [2].
In a broader taxonomic sense, the Myrtaceae is
Address for correspondence
Mahendra K R
Email: mahigene8611@gmail.com
49 Internationale Pharmaceutica Sciencia
the rose and mallow families. The Myrtaceae falls into
the order Mytrales, along with the families Lythraceae,
Punicaceae (pomegranates), and Onagraceae (evening
primroses). There are 121 genera, comprising about
3850 species, occurring in tropical and warm regions
of Myrtaceae Family, and abundant in Australia [3].
Natural products have inspised many
developments in oranic chemistry leading to advances
in synthetic methodologies in developing several
analogues of lead compounds with therapeutical
potential. Acceptance of Cinchona for the treatment of
malaria in the 17th century, followed by digitalis,
morphine etc, and the introduction of aspirin, brought
the general public to believe in the wonders of diverse
floral wealth, and that natural products including
plants offer large structural diversity for
pharmacological treatment of various classes of
disorders [4].
In the present study we are analyzing the
phytochemicals present in some plant leaves of
Myrtaceae Family (Guava, Jambul, and Eucalyptus)
Jul-Sep 2012 Vol 2 Issue 3
 Mahendra K R et al: HPLC Analysis of Methanolic Extract of Some Medicinal Plant Leaves of Myrtaceae Family
through the chromatographic technique. Plants are
always used for curing the disease from the ancient
time hence it is important to analysis of chemical
present in plant parts.
Guava (Psidium guajava L.), native to tropical
America, is a tree which is now widely distributed
throughout the tropic and subtropic areas, and its
fruits are consumed fresh or processed. The major
producers of guava are South Africa, India, Hawaii,
Colombia, Puerto Rico, Jamaica, Brazil, and Israel.
Leaves and fruits of guava showes antioxidant activity,
free radical-scavenging effects and antidiabetic activity
[5, 6, 7, 8].
Jambul (Syzygium cumini) is
to Bangladesh, India, Nepal, Pakistan, Sri Lanka, the
Philippines, and Indonesia. It possesses
biological activities such as antioxidant, antidiabetic
and antibacterial [9, 10, 11, 12, 13, 14].
Eucalypts are highly valued in
communities for a wide range of uses. These include
fuel, poles, small lumber and furniture, essential oils
and tannins. The main species grown include
Eucalyptus camaldulensis, E. tereticornis,
urophylla and E. grandis in subtropical and tropical
regions, and E. globules in more temperate climes.
Leaves extract of Eucalyptus have possess different
biological activity such as antidiabetic, antioxidant and
antibacterial [ 15, 16,17 ].
Material and Methods
Sample Collection
The plant leaves of Psidium guajava,
Syzygium cumini, E. globules were collected and
identified in march 2012 from the campus of Mahatma
Gandhi Chitrakoot Gramoday Vishwavidyalaya, Satna
(M.P.). All plant leaves were washed with fresh water
and dried under the shade at room temperature. The
leaves were powdered and stored in sterile container
for further use.
Preparation of Plant Extracts
1 gm of fine powdered sample of plant leaves
were extracted with 10 ml HPLC grade methanol
through open reflux process at 40 °C for 30 min. Then
Internationale Pharmaceutica Sciencia Jul-Sep 2012
the extracts were filtered through filter paper
(Whatman No. 1) to remove free unextractable
substances. The filtrates of plant extracts were
preserved 4-5°C for further process.
Experimental Methods
HPLC Analysis
The HPLC analysis of methanolic extract was
carried out with Chromatographic system (YL 9100,
Korea) consist of autosampler (YL 9150) with 100 µl
fixed loop and an YL9120 UV-Visible detector. The
separation was performed on a SGE Protecol
PC18GP120 (250mm×4.6 mm, 5µm) column at
native ambient temperature. The mobile phase consists of
methanol to water (70:30 v/v) and the separations
many were performed by using isocratic mode, elution
performed at a flow rate of 1 ml/min. The samples
were run for 15min. and detection was done at 254 nm
rural by UV detector. All chromatographic data were
recorded and processed using autochro-3000 software.
Results and Discussion
E. Standardization and characterization of herbal
drugs is a topic of continuous scientific interest in the
herbal drug industry. With the advent of modern
chromatographic systems there is an ever increasing
intent to produce and develop easy, rapid, convenient
and cost effective methods for standardization [18].
For standardization of methanolic extract of
plant leaves, HPLC is a sensitive and accurate tool that
widely used for the quality assessment of plant extract
and its derived product/formulation [19]. HPLC
fingerprints of Psidium guajava, Syzgium cumini and
Eucalyptus are given in Figure 1-3.
Results of HPLC analysis (Figure-1) of
Psidium guajava methanolic extract, at 254 nm,
shows presence of various constituents as evidenced by
the chromatogram obtained at various retention times
( 1.6833, 2.5000, 3.2167, 3.6333, 3.9833) are the
constituents found in Psidium guajava leaves mainly.
The methenolic extract of Syzygium cumini
leaves extract chromatogram (Figure-2) shows
different constituents at various retention times
Vol 2 Issue 3 50
 Mahendra K R et al: HPLC Analysis of Methanolic Extract of Some Medicinal Plant Leaves of Myrtaceae Family

(1.5833, 2.4333). These two peaks are represents the
main constituents present in the plant leaves.
Similarly in the methanolic extract
Eucalyptus leaves (Figure-3) show the various
constituents with different retention times (2.5500,
3.1393, 3.5000, 4.9000, 5.6333, and 7.0000). In which
the compound having RT 2.5500 is the main
constituents in methanolic leaf extract.
In the present study, we have described a
method that was developed to profile methanol soluble
Figure-1: HPLC chromatogram of methanolic extract of Psidium guajava L. leaves at 254 nm
phenols, which are mainly flavonoid glycosides which
show the intense absorption in UV region [20]. This
of technique provides us chromatogram of different
compounds present in methanolic extract of plants. In
the absence of authentic standards these UV spectra
only allow determination of compound class.
Therefore, this method must be coupled to HPLC-MS
and/or NMR analyses in order to identify completely
the compounds detected and get some insight into
there structure.

Figure-2: HPLC chromatogram of methanolic extract of Syzygium cumini leaves at 254 nm

Figure 3: HPLC chromatogram of methanolic extract of Eucalypts leaves at 254 nm

51 Internationale Pharmaceutica Sciencia Jul-Sep 2012 Vol 2 Issue 3

 Mahendra K R et al: HPLC Analysis of Methanolic Extract of Some Medicinal Plant Leaves of Myrtaceae Family

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