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Asoso et al. Antibacterial screening and GC-MS prole pp. 19-25
e - ISSN: 2602-8360 - Volumen. 2, Nº 2, Junio – Noviembre 2018
INTRODUCTION
The World Health Organizaon (WHO) denes
tradional medicine as the sum of total of the
knowledge, skills, and pracces based on the
theories, beliefs and experiences indigenous to
dierent cultures, whether explicable or not,
used in the maintenance of health as well as
in the prevenon, diagnosis, improvement or
treatment of physical and mental illness (1).
The use of herbal medicine as alternave therapy
has prevalent throughout the world due to the
growing resistance of pathogens to convenonal
anbiocs (2). The need for more potent, safe
and aordable drugs has led to intensied
research into herbal drugs, the result of which
is the introducon of new herbal preparaon
for therapeuc uses (3).Medicinal plants are
frequently used as remedies for many infecous
diseases (4). The treatment and control of
diseases by the use of the available medicinal
plants in a locality have been helpful and of a
priority to majority of urban and rural dwellers in
healing various diseases because of the reliability
and stability in plant products for healing (5).
Calotropis procera (Apple of Sodom) is a shrub or
small tree, which has become a serious weed in
pastures and overgrazed rangelands. It is a nave
to West Africa as far as south as Angola, North and
East African, Madagascar, the Arabian Peninsula,
Southern Asia, India and China to Malaysia.
Calotropis was formerly placed in the family of
Asclepiadaceous (the milkweed family), which is
now considered a subfamily of the Apocynaceae
(6).
MATERIALS AND METHOD
Collecon of plant samples
Apparently healthy plant namely Calatropis
procera were collected from Ado-Eki, Eki State
Nigeria.
The plants parts leave and stem were air-dried
for 5 weeks at room temperature (25 + 2 oC) and
then ground to powder with a mechanical grinder
(Thomas Wiley machine, model 5 USA). Powders
(200gs) of each plant were extracted with 1litre
of sterile aqueous water, ethanol, methanol and
acetone separately at room temperature (25 + 2
oC). They were labeled as crude extracts.
Anbacterial Screening of the Crude Fracons
The evaluaon of anmicrobial acvity was
performed for all fracons by the paper disc
agar diusion method following the standard
rules of anmicrobial sensivity tests by the
Clinical and Laboratory Standards Instute (7).
The anbacterial eect of the fracons was
carried out whereby the discs were previously
impregnated with the plant crude fracons and
placed on the sterile prepared medium. The
plates were incubated at 37 oC for 24 - 48 hours.
The sensivity of the test organisms to each of
the extracts was indicated by clearing around
each disc. The diameter as an index of the degree
of sensivity was measured with a transparent
plasc ruler.
Column chromatography and fracon extracts
Glass wool was placed at the outlet of a column
aer which, one gram of plant extract powder
was weighed into it and subjected to column
chromatography (30 x 8 cm column) using
60 g of silica gel 60 F254 (Merck, 0.020 mm
thickness). The column was successively eluted
rst with petroleum ether (150 ml) and then with
chloroform (80 ml): methanol (2 ml) (40:1) and
nally with 100% methanol (150 ml). Each 100
ml eluent was collected into a round boom ask
(250 ml) capacity and dislled to obtain fracons.
The fracons collected were numbered. GC-MS
(Gas Chromatography-Mass Spectrophotometry)
analysis was carried out in GCMS-QP2010 PLUS
Shamadzu.
Anbacterial screening of crude fracons
The anbacterial screening of the crude fracons
of Calotropis procera leaf was examined; some
chemicals were used in the fraconalizaon of the
crude sample, which are methanol, chloroform
and petroleum ether. Methanol, aqueous and
acetone leaf fracons showed the highest zone of
inhibion were: 30, 20 and 20 mm respecvely.
The use of petroleum ether do not show any
signicant anbacterial eects against the
isolates while aqueous crude fracons shows an
eecve anbacterial acvity against S. aureus at
zone of inhibion of 15 mm (Table 1).