The document summarizes a study that investigated the phytochemical composition and antimicrobial activity of dried seed extracts of Maranthes polyandra. Key findings include: 1) Preliminary phytochemical screening revealed the presence of steroidal triterpenes, cardiac glycosides, and trace amounts of alkaloids in the chloroform, ethyl acetate, and methanol extracts. 2) The ethyl acetate extract exhibited the highest inhibition zone (29mm) against Candida stellatoidea. Ethyl acetate and methanol extracts inhibited growth of many test organisms at a concentration of 3.75mg/mL. 3) The minimum bactericidal/fungicidal concentration of most extracts against

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PRELIMINARY PHYTOCHEMICAL SCREENING
AND ANTIMICROBIAL ACTIVITY OF DRIED SEED
EXTRACTS OF MARANTHES POLYANDRA
ARTICLE · APRIL 2015
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T. A. Tor-Anyiin
Federal University of Agriculture, Makurdi
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John Anyam
Ahmadu Bello University
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Jecinta Ndiombueze Anowu
Federal University of Agriculture, Makurdi
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Available from: John Anyam
Retrieved on: 11 October 2015

2. J. Chem.Soc.Nigeria, Vol. 40,No.1, pp 24-27[2015]
PRELIMINARY PHYTOCHEMICAL SCREENING AND
ANTIMICROBIAL ACTIVITY OF DRIED SEED EXTRACTS OF
MARANTHES POLYANDRA
* T. A. Tor-Anyiin, J. V. Anyam, G. Anger and J. N. Anyam
Phytochemistry Research Group, Department of Chemistry, University of Agriculture, Makurdi,
Nigeria.
Accepted: 07/04/2015
*
Corresponding author:toranyiint@yahoo.com
Abstract
The chloroform, ethyl acetate and methanol extracts of dried seeds of Maranthes polyandra were
subjected to preliminary phytochemical screening. The extracts revealed presence of steroidal
triterpenes, cardiac glycosides and trace of alkaloids. Antimicrobial activities of the extracts were carried
out using broth dilution method. The test organisms were laboratory isolates of Staphylococcus aureus,
Streptococcus pyogenes, Escherichia coli, Salmonella typhi, Shigella dysenteriae, Klebsiella pneumoniae,
Candida albicans, Candida krusei, Candida tropicalis, Candida stellatoidea, Microsporum gypseum,
Microsporum sp. and Trichophyton rubrum. Ethyl acetate extracts exhibited the highest zone of inhibition
(29 mm) against Candida stellatoidea. Ethyl acetate and methanol extracts inhibited growth of many
tested organisms at concentration of 3.75 mg/mL. The minimum bactericidal/fungicidal concentration of
other extracts against the test organisms was 15 mg/mL while ethyl acetate was 7.5 mg/mL against
Staphylococcus aureus, Salmonella typhi, Shigella dysenteriae, and Candida stellatoidea. This study has
justified the traditional use of the plant for treatment of various diseases that are caused by these
pathogens.
Key words: Phytochemical analysis, antimicrobial activity, seed extracts, Maranthes polyandra
Introduction
The development of resistance of pathogens
to synthetic as well as side-effects of vended
antibiotics has justified continuing search for anti-
pathogenic agents from natural sources. Since ancient
times, man has looked for cure from Nature. Despite
the great advances made in modern medicine, plants
still have an important contribution to health care [1].
Natural products have continued to serve as very
fruitful source of drug leads especially when coupled
with screening methods based on modern
pharmacological or biochemical products [2].
Bacterial and fungal infections are among the most
dangerous and opportunistic diseases to vulnerable
people [3]. Maranthes polyandra, (Benth) Prance,
Chrysobalanacae (Tiv: Ibua-kuna;Hausa: Gongea-
kussa;Yoruba:Abere; Ghana: (Akan-asante:Abrabes);
Senegal: tudu, Togo: bende-moso)is a common tree
in the Guinea Savanna. It is an evergreen flowering
tree producing dark red or black-purple ellipsoid
fruits to 2.5 cm long or less. The seeds contains a lot
of oil but are not considered edible [4].
Maranthes polyandra is a plant widely used
in traditional medicine for a number of ailments. The
bark is used for fever, crushed leaves in treatment of
fractures and powdered roots for curing syphilis [4].
This study was therefore undertaken to screen
phytochemically for secondary metabolites in the
root back extracts of this plant as well as the
antimicrobial properties of the extracts. This has
become inevitable against the backdrop of increase
recourse to seeking medical treatment through
traditional medicine and in addition to seeking anti-
infective agents from natural sources.
Materials and Methods
Plant seeds were collected at Sev-Av, 7o
52N, 9o
37E,
Mbawar, Wukari Local Government Area, Taraba
State of Nigeria in September 2013. The plant was

3. T. A. Tor-Anyiin, J. V. Anyam, G.Anger and J.N. Anyam
2
identified by Mr. Ikyobo John N., Department of
Wildlife and Range Management, University of
Agriculture Makurdi. Seeds were air dried for two
weeks, size reduced in a wooden mortar and
packaged in a polythene bagwith appropriate label.
Dried pulverized seeds (600 g) were defatted with
500 mL of n-Hexane for 72 hours at ambient
temperature and filtered. The marc was sequentially
treated to microwave assisted extraction using 500
mL each of chloroform, ethyl acetate and methanol.
Extracts were filtered and excess volume reduced in
vacuo using rotary evaporator at 50 o
C, and left to
evaporate to dryness at room temperature.
Phytochemical screening
Crude extracts were subjected to phytochemical tests
for presence of saponins, tannins, flavonoids,
anthraquinones glycosides, steroids, cardiac
glycosides, and alkaloids, using standard procedures
[5, 6].
Antimicrobial Activities
Antimicrobial activities of the plant extracts were
determined using some pathogenic microbes. An
extract (0.3 g) was weighed and dissolved in DMSO
(10 mL) to give a concentration of 30 mg/mL. This
served as initial concentration of extract used to
determine antimicrobial activities of the extracts.
Mueller Hinton and Sabourand dextrose agar were
the growth media used for bacteria and fungi,
respectively. All media were prepared according to
the manufacturer instructions, sterilized at 120 o
C in
an autoclave for 15 minutes, poured into sterile petri-
dishes and allowed to cool and solidify. Diffusion
method was used for screening of extracts. Mueller
Hinton agar was seeded with standard inoculum (0.1
mL) of bacteria and Sabourand dextrose agar with the
fungi. The inocula were spread evenly over the
surface of the media with a sterile swab. By use of a
standard cork borer of 6 mm in diameter, a well was
cut at the centre of each inoculated medium. Solution
of the extract (0.1 mL) of concentration 30 mg/mL
was then introduced into each well on the medium.
Inoculated plates were incubated at 37 o
C for 24 hrs
for the bacteria and at 30 o
C for 7 days for the fungi,
after which each plate was observed for inhibition
zone of growth. Zones were measured with a
transparent ruler and the result recorded in
millimeters.
The minimum inhibition concentration (MIC) of an
extract was carried out using broth method. Mueller
Hinton and Sabourand dextrose broth were prepared,
10 mL of the broth was dispensed into test tube and
sterilized at 121 o
C for 15 minutes, then allowed to
cool.
McFarland’s turbidity standard scale number 0.5 to
give turbidity solution was prepared. Normal saline
was prepared, 10 mL was dispensed into sterile test
tube and the test microbe was inoculated and
incubated at 37 o
C for 6 hours. From there dilution of
the test microbe was done in the normal saline until
the turbidity matched that of the McFarland’s scale
by visual comparison. At this point, this
test microbe had a concentration of about 1.5 x 108
cfu/mL. A two-fold serial dilution of the extract in
the sterilized broth was made to obtain the
concentration of 30, 15, 7.5, 3.5 and 1.875 mg/mL,
respectively. Initial concentration was obtained by
dissolving 0.3 g of extract in 10 mL of sterile broth.
Having obtained the different concentrations of the
extract in the sterile broth 0.1 mL of the test microbe
in the normal saline was then inoculated into the
different concentrations. Incubation was made at
37o
C for 7 days for the fungi, after which the broth
was observed for turbidity growth. Lowest
concentration of extract in the broth which showed
no turbidity was recorded as the MIC.
Minimum bactericidal concentration/Minimum
fungicidal concentration (MBC/MFC) were carried
out to determine whether the test microbes were
killed or only their growth was inhibited. Mueller
Hinton and Sabourand dextrose agar were prepared
and sterilized at 121 o
C for 15 minutes then poured
into sterile petri-dishes and allowed to cool and
solidify. The content of the MIC in the serial dilution
were sub-cultured into the prepared media,
incubation were made at 37 o
C for 24 hours for
bacteria and at 30 o
C for 7 days for fungi, after
which plates were observed for colony growth.
MBC/MFC were the plates with lowest concentration
of extract without colony growth.
Results and Discussion
Phytochemistry
Phytochemical screening of crude chloroform, ethyl
acetate and methanol extracts of Maranthes
polyandra revealed presence of, steroidal triterpenes,
cardiac glycosides and alkaloids in trace amounts but
saponins glycosides, tannins, flavonoids,
anthraquinones, free anthraquinones and
carbohydrates were found to be absent (Table 1).
Table 1: Phytochemical screening for various
extracts of Maranthes polyandra

4. T. A. Tor-Anyiin, J. V. Anyam, G.Anger and J.N. Anyam
3
The antimicrobial screening (Table 2) showed the
susceptibility test against grams positive and negative
organisms and fungi species.The extracts showed
inhibition activity against some of the test organisms.
The antimicrobial activity shown by these extracts is
due to secondary metabolites present in the plant. The
observed properties of this plant correlate with the
oral information given by the indigenous tribe or
community that use this plant for basic health care.
Ethyl acetate extract exhibited highest considerable
level of inhibition against C.stellatoidea(29 mm) and
chloroform extract exhibited least against
C.tropicalis (24 mm).
Keys: +ve = positive and –ve = negative
These classes of compounds are known to show
curative activity against several pathogens and
therefore could explain its use traditionally for the
treatment of a variety of diseases.
Table 2: The Antimicrobial Activities of Maranthes polyandra
Pathogen Sensitivity/Zone of Inhibition (mm)
Crude Extract Control(5µg/ml)
Chloroform
Ethylacetate
Methanol
Erythromycin
Ciprofloxacin
Fulcin
Fluconazole
S. aureus S/24 S/27 S/25 S/27 S/35 R/0 R/0
S. pyogenes S/25 S/26 S/25 S/32 S/29 R/0 R/0
E. coli R/0 R/0 R/0 R/0 S/35 R/0 R/0
S. typhii S/24 S/28 S/26 R/0 S/37 R/0 R/0
S. dysenteriae S/26 S/27 S/26 S/40 S/41 R/0 R/0
P. aeruginosa R/0 R/0 R/0 S/29 S/35 R/0 R/0
K. pneumoniae R/0 R/0 R/0 S/35 R/0 R/0 R/0
C. albicans S/24 S/27 S/26 R/0 R/0 R/0 S/32
C. tropicalis S/24 S/25 S/24 R/0 R/0 R/0 S/30
C. thrusei R/0 R/0 R/0 R/0 R/0 R/0 S/35
C. Stellatoidea S/25 S/29 S/26 R/0 R/0 R/0 S/30
M. gypseum R/0 R/0 R/0 R/0 R/0 S/32 R/0
Microsporum spp R/0 R/0 R/0 R/0 R/0 S/30 R/0
T. rubrum S/24 S/26 S/25 R/0 R/0 S/37 R/0
Keys: S = Sensitive, R = Resistant
Phytochemicals Results for various
extracts
Cf EA Meth
Reducing sugars -ve -ve -ve
Free Anthraquinones -ve -ve -ve
Anthraquinone
Glycosides
-ve -ve -ve
Saponin Glycosides -ve -ve -ve
Steroids +ve +ve +ve
Terpenes -ve +ve +ve
Tannins -ve -ve -ve
Alkaloids -ve -ve +ve
Cardiac glycosides -ve +ve +ve
Flavonoids -ve -ve -ve

5. T. A. Tor-Anyiin, J. V. Anyam, G.Anger and J.N. Anyam
4
Table 3: Minimum Inhibition Concentration of the Extracts
Pathogen Chloroform Ethyl acetate Methanol
30mg/ml
15mg/ml
7.5mg/ml
3.5mg/ml
1.875mg/ml
30mg/ml
15mg/ml
7.5mg/ml
3.5mg/ml
1.875mg/ml
30mg/ml
15mg/ml
7.5mg/ml
3.5mg/ml
1.875mg/ml
S. aureus - - # + α - - - # + - - # + α
S.pyogenes - - # + α - - - # + - - # + α
S. typhi - - # + α - - - # + - - - # α
S.dysenterteriae - - - # + - - - # + - - - # α
C.albicans - - # + α - - - # + - - - # α
C.tropicalis - - # + α - - # + α - - + + α
C. stellatoidea - - # + α - - - # + - - - # α
T.rubrum - - # + α - - - # + - - # + α
Key: - = No turbidity (No growth), # = Minimum Inhibition Concentration (MIC), + = Turbidity (light growth), α = Moderate turbidity
From Table 3 it was observed that, the activities (i.e.
minimum inhibition) of the seeds extracts of
Maranthes polyandra against the test organisms was
noticed at concentration of 7.5 mg/mLfor almost all
the extracts, except ethylacetate extract which has
this value noticed only on C.tropicalis.
C.albicans in ethylacetate and methanol extracts with
MBC/MFC values of 7.5 µg/mL. There are several
reports on antibacterial properties of plants used in
trado-medicine by various cultures/communities [7].
The higher antibacterial and antifungal activities of
these
Table 4: Minimum Bacteriadal/Fungicidal Concentration of Extracts against Test Microorganisms.
Germ PE CF EA METH
30mg/ml
15mg/ml
7.5mg/ml
3.5mg/ml
1.875mg/ml
30mg/ml
15mg/ml
7.5mg/ml
3.5mg/ml
1.875mg/ml
30mg/ml
15mg/ml
7.5mg/ml
3.5mg/ml
1.875mg/ml
30mg/ml
15mg/ml
7.5mg/ml
3.5mg/ml
1.875mg/ml
S. a - # + α И - # + α И - - # + α - # + α И
S.p - # + α И - # + α И - # + α И - # + α И
S. t - # + α И - # + α И - - # + α - # + α И
S.d - # + α И - # + α И - - # + α - # + α И
C.a - # + α И - # + α И - - # + α - - # + α
C.t - # + α И - # + α И - # + α И - # + α И
C. s - # + α И - # + α И - - # + α - # + α И
T.r - # + α И - # + α И - # + α И - # + α И
Key:- = no colony growth, # = MBC/MFC, + = scanty colony growth, α = moderate colony growth and И = heavy colony growth; S.a=S. aureus,S.p=S.
pyogenes,E.c=E. coli,S.t=S. typhii,S.d=S. dysenteriae,P.a=P. aeruginosa,K.p=K. pneumoniae,C.a=C. albicans,C.t=C. tropicalis,C.th=C. thrusei,C.s=C. Stellatoidea,M.g=M.
gypseum,Ms=Microsporum spp,T.r=T. rubrum

6. T. A. Tor-Anyiin, J. V. Anyam, G.Anger and J.N. Anyam
5
Minimum bactericidal/fungicidal
concentration of the seeds extracts of
Maranthes polyandra presented in Table 4,
indicated that, the test organisms could be
inhibited at concentration of 15 µg/mL except
S.aureus, S. typhi, S. dysenteriae, and
C.stellatoidea in ethylacetate extract and
extracts could not be unrelated to the
presence of the plant secondary metabolites
detected.
Conclusion
The extracts especially ethylacetate exhibited
considerable inhibition activities against
almost all the test organisms and comparable
activities to the reference drugs under study;
thus providing somescientific basis for some
of the folkloric claims. The isolation and
possible characterization of the bioactive
constituents from the extracts of this plant
species as possible antimicrobial agent is
recommended.
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