International Journal of Scientific Research in Chemical Engineering, 1(2), pp.

17-23, 2014
Available online at http://www.ijsrpub.com/ijsrce
ISSN: 2345-6787; 2014 IJSRPUB
http://dx.doi.org/10.12983/ijsrce-2014-p0017-0023
17
Full Length Research Paper
Qualitative Evaluation of Phytochemical Composition, Antioxidant and
Antimicrobial Activities of Extracts of Coral Tree - Erythrina Senegalensis DC
Oluremi Isola Adeniran*
1
, Sunday Felix Abimbade
2
1
Department of Chemistry, Faculty of Science, University of Abuja, Nigeria
2
Department of Industrial Chemistry, Federal University, Oye Ekiti, Nigeria
*Corresponding Author: benadole@yahoo.com; Tel: 08027263867
Received 26 March 2014; Accepted 19 May 2014
Abstract. The inability of a majority of world population to afford the cost of modern healthcare is a serious problem that calls
for careful research into health promoting components of plants and urgent need to tap from the potentials of their molecular
diversities. This study aimed at investigating some properties of Erythrina senegalensis, a plant with several uses in traditional
medicine. The n-hexane, ethyl acetate and ethanol extracts of each of the leaves, stem bark and root bark of the plant were
investigated for their phytochemical composition, antioxidant properties and antimicrobial potentials on some pathogens. The
phytochemical analysis revealed the presence of some pharmacologically important molecules such as, alkaloids, cardiac
glycosides, flavonoids and steroids in one or more parts of the plant that justify some of its ethno-medicinal applications. All
the extracts of each plant part show considerable antioxidant activity, but, the root bark extracts showed highest activity having
values ranging from 82.8 to 91.5% and 78.9 to 94.6% at the lowest (0.05 mg/ml) and optimum (0.5 mg/ml) concentrations
respectively. The antimicrobial susceptibility test revealed zones of inhibition 9 mm for extracts with activity while the stem
bark extracts in its entirety showed no activity. Altogether, the plant has very narrow spectrum of activity.
Keywords: Erythrina senegalensis, qualitative evaluation, antioxidant, antimicrobial, phytochemical
1. INTRODUCTION
The use of plant in Nigeria traditionally for curative
purposes should be a sustained practice due to the fact
that a large majority of her population cannot afford
the cost of modern healthcare. Coupled with this is the
fact that the likelihoods of making new findings
become obvious considering that a single plant may
contain up to thousands of constituents; investigation
of plants thus become necessary for better
understanding of their properties, efficacy and safety
(Banso and Adeyemo, 2007; Nascimento et al., 2000;
Hostettmann et al., 1995). On these notes Erythrina
senegalensis, a plant used in traditional medicine to
cure many diseases but with several of these uses yet
to be scientifically explored, has been chosen for
investigation.
Erythrina senegalensis DC belonging to the family
Fabaceae is a prickly shrub with bright red flowers
(Doughari, 2010; Trease and Evans, 2002). Some of
its common names among some ethnic groups in
Nigeria include coral flower and parrot tree (English),
murjiya (Hausa), echichi (Igbo), and ologun sheshe
(Yoruba) (Blench, 2007; Udem et al., 2009). Erythrina
is a genus of flowering plants containing about 130
species, which are widespread in tropical and
subtropical regions universally (Trease and Evans,
2002). This generic designation probably stems from
the Greek word erythros, meaning "red" and denoting
the flower colour many species have (Saidu et al.,
2000). E. senegalensis is a tree 7-15 m tall. The
flowers, bright red and 4-5 cm in length occur in large
groups at the end of the twigs. The leaves are
composed of three leaflets, each having a surface area
between 20 and 150 cm
2
and borne on a thorny stalk.
The fruit comprises a slightly hairy warped pod, 7-15
cm long (Doughari, 2010; Trease and Evans, 2002;
Sofowora, 1993). Coral tree is native to Senegal,
Nigeria, Mali, Liberia, Cameroon and Benin
(Akinpelu and Onakoya, 2006; Sofowora, 1993). It is
a plant that has a large number of traditional medicinal
uses in West Africa including treatment of
amenorrhea, liver disorders, kidney problems, dropsy,
epilepsy, venereal diseases and prevention of abortion
(Burkill, 1985).
Despite its numerous uses, there is, however,
dearth of information on the scientific evidence for
most of the usages of coral tree in traditional
Adeniran and Abimbade
Qualitative Evaluation of Phytochemical Composition, Antioxidant and Antimicrobial Activities of Extracts of Coral
Tree - Erythrina Senegalensis DC
18
medicine. This study therefore aims at investigating
the plants composition, antioxidant and anti-
microbial activities with a view to tapping from its
health-promoting abilities for enhancement of human
wellbeing.

2. MATERIALS AND METHODS

2.1. Collection and preparation of samples

Fresh samples of Erythrina senegalensis (Leaves,
stem bark, and root bark) were collected from Sheda,
Kwali Area Council, Federal Capital Territory (FCT),
Abuja, Nigeria. Sample identification and
authentication were carried out at Botany Unit,
Department of Biological Science, University of
Abuja. The samples were washed, air-dried and
ground into powder.

2.2. Extraction of samples

About 500 g of each of the powdered samples was
subjected to gradient extraction with hexane, ethyl
acetate and ethanol successively, in a Soxhlet
extractor. Each extract so obtained was concentrated
using rotatory evaporator followed by evaporation to
dryness on water bath. The dry extracts were stored
for further analysis.

2.3. Phytochemical screening

The extracts were subjected to a blend of
phytochemical tests described by Debela (2002) and
Trease and Evans, (2002). Presence of alkaloids was
confirmed by a reddish-brown precipitate when 0.5 g
of extract is stirred with 5 ml of 1% aqueous HCl on
water bath and then filtered followed by addition of 1
ml of Wagners reagent to 1 ml of the filtrate.
Appearance of a pink, red or violet colour in the
ammoniacal phase confirms the presence of free
anthraquinones on addition of 0.5 ml ammonia
solution to the filtrate obtained after shaking 0.2 g of
extract with 10 ml of benzene and filtering. On
dissolution of 0.5 g of extract in 2 ml glacial acetic
acid containing 1 drop of ferric chloride solution
under-laid with 2 ml of concentrated sulphuric acid, a
brown ring formation at the interphase indicates the
presence deoxy-sugar characteristic of cardiac
glycosides. Presence of flavonoids is detected by
decolourization of a yellow solution on addition of
concentrated HCl to a solution of the extract in dilute
NaOH. When 0.5g of the extract was added to 1%
ferric chloride in 50% aqueous methanol, a dirty green
precipitate indicates the presence of phenols. A
reddish precipitate formed when a few drops of 1%
HCl were added to 1ml of extract and boiled indicates
the presence of phlobatannins. Subjection of the
extract to frothing test as a preliminary test and to
haemolysis test for confirmation was carried out for
detection of saponin. When a well shaken mixture of
extract, chloroform and few drops of conc. H
2
SO
4
is
allowed to stand for some time, appearance of red
colour at the lower layer indicates the presence of
steroids. To test for tannins, about 0.5 g of extract was
stirred with 10 ml of distilled water, filtered, and few
drops of 1% ferric chloride solution subsequently
added to 2 ml of the filtrate. The presence of tannins is
indicated by a blue-black, green or blue-green
precipitate. The formation of a turbid brown colour
shows the presence of terpenoids when 2 ml of
benzene was added to 1 ml of extract.

2.4. Determination of antioxidant activity

The free radical scavenging activities of the plant
extracts against 2, 2-diphenyl-1-picryl-hydrazyl
radical (DPPH) were determined according to
methods of Brand-Williams et al. (1995) and Mensor
et al. (2001) with a little modification. Different
concentrations (0.05, 0.1, 0.5, 1.0, 2.0 and 5.0 mg/ml)
of each of the nine extracts were prepared in
methanol. To various concentrations of each of the
test extracts was added 0.5 ml of 1 mM of DPPH. The
resultant mixture was allowed to stand for about 30
minutes in the dark at room temperature after which
the reading of absorbance was taken using UV-Visible
spectrophotometer at 517 nm. Vitamin C was used as
the antioxidant standard (positive control) at
concentrations of 0.05, 0.1, 0.5, 1.0, 2.0 and 5.0
mg/ml while a blank solution was prepared containing
the same amount of methanol and DPPH (negative
control).
Percentage antioxidant activity or percentage
inhibition of the extracts was calculated using the
formula:

Where: A
b
= Absorbance of blank (without extract); A
a
= Absorbance of test extract

2.5. Test for antimicrobial activity

The isolates (test bacteria) were seeded onto the
nutrient agar plates by swabbing using a sterile cotton
swab for each organism. Disc diffusion method was
used to determine the antibacterial activity of the plant
extracts. The papers were cut into small sizes and
introduced into each sample bottle containing the
International Journal of Scientific Research in Chemical Engineering, 1(2), pp. 17-23, 2014
19
diluted extracts. They were dried at 50
o
C and used in
determining zone of inhibition. The sterilized discs
incorporated with the extracts were placed on the
nutrient agar plates that were seeded with the test
bacteria and incubated. The set-up was then incubated
for 24 hours at 37
o
C. The zones of inhibition were
measured.

3. RESULTS

Phytochemical constituents of extracts of E.
senegalensis Percentage Antioxidant Activity of
extracts of coral tree and standard have been shown in
tables 1 and 2. The effect of the hexane, ethyl acetate
and ethanol extracts of E. senegalensis (Leaves, Stem
bark and root bark) on some test bacteria revealed the
following results as shown in Table 3.







Table 1: Phytochemical constituents of extracts of E. senegalensis

Key: + = present; - = absent; Hex = hexane; EtAc = ethyl acetate; EtOH = ethanol


Table 2: Percentage Antioxidant Activity of extracts of coral tree and standard

Key: Hex = Hexane; EtAc = Ethyl acetate; EtOH = Ethanol; Vit. C = Vitamin C Standard.


Table 3: Antimicrobial activity of the Leaves, stem bark and root bark extracts E. senegalensis (Zones of inhibition are shown
in mm)

Key: NI = No inhibition, + = 9 mm, Hex = Hexane, EtAc = Ethyl acetate, EtOH = Ethanol

Adeniran and Abimbade
Qualitative Evaluation of Phytochemical Composition, Antioxidant and Antimicrobial Activities of Extracts of Coral
Tree - Erythrina Senegalensis DC
20
4. DISCUSSION

Qualitative phytochemical investigation of the plant
extracts revealed that alkaloids and saponins were
present in all the plant parts leaves, stem bark and
root bark- (Table 1). The coexistence of both alkaloid
and saponin in a plant has been reported to be
responsible for the antihypertensive property of such a
plant (Fahey, 2005; Krishnaiah et al., 2009). This
activity is attributed to ability of saponin to prevent
excessive intestinal absorption of cholesterol (Olaleye,
2007; Akinpelu and Onakoya, 2006; Raffauf, 1996).
The presence of anthraquinones and cardiac
glycosides in stem bark and root bark respectively
may be the reason for the use of the plant as a laxative
(Burkill, 1985) and treatment of some heart related
problems (Togola et al., 2005 ). Occurrence of
flavonoids especially in the leaves and root bark of the
plant may be a justification for its use in treatment of
diarrhoea. Schuier et al. (2005), credited the ability of
a plant to treat diarrhoea to the capability of
flavonoids to inhibit the development of fluids that
cause diarrhoea. On the other hand, it has been shown
that cardiac glycosides prevent cardiovascular
diseases by inhibiting the sodium ions-potassium ions
pump thereby increasing the level of sodium ions in
the myocytes, which then leads to a rise in the level of
calcium ions available for contraction of the heart
muscle, consequently improving cardiac output and
reducing heart distension. The antioxidant effect of
the extracts was evaluated on DPPH free radicals with
vitamin C as reference. All the extracts except root
bark hexane (RHex) extract showed dose-dependent
antioxidant activity at lower concentrations (from 0.05
mg/ml to 0.5 mg/ml) and a steady decline in activity at
higher concentrations (from 0.5 mg/ml to 5 mg/ml).
The exact opposite trend was observed for the root
bark hexane extract (Table 2 and Fig. 1).


Fig. 1: Antioxidant Activity of the Extracts of E. senegalensis
Key: LHex = Leaves hexane extract; LEtAc = Leaves ethyl acetate extract; LEtOH = Leaves ethanol extract;
SHex = Stem bark hexane extract; SEtAc = Stem bark ethyl acetate extract; SEtOH = Stem bark ethanol extract;
RHex = Root bark hexane extract; REtAc = Root bark ethyl acetate extract; REtOH = Root bark ethanol extract.

Previous reports on the antioxidant activity of this
plant (Atsamo et al., 2013; Njayou et al., 2010;
Donfack et al., 2008) are mostly on the stem bark
extracts; the present study is about the first to report
the antioxidant activity of the root bark of the plant.
Although each of the extracts showed appreciable
antioxidant activity, the root bark extracts in general
compare favourably with the reference and the root
bark ethyl acetate (REtAc) extract in particular
exhibited activity (ranging from 91.0 to 94.6%) in
high proximity with the standard used (in the range
96.9 to 98.4%). It is obvious from the foregoing
evidence that these extracts have potentials of serving
as substitute for known standard antioxidants such as
ascorbic acid (Vitamin C).
The entire stem bark extracts as well as the leaf
hexane extract and root bark ethyl acetate extracts
showed no antimicrobial activity against the test
organisms and where there is activity the zone of
inhibition is less than 9 mm (Table 3). The ethyl
acetate extract of the leaf showed a weak activity
against only Proteus species while the ethanol extract
of the same was active against Proteus spp and S.
aureus. Similarly ethyl acetate extract of the root bark
showed activity against S. aureus and B. subtilis while
its ethanol extract inhibits S. aureus, B. subtilis and
Proteus spp. It could be inferred from the assessment
International Journal of Scientific Research in Chemical Engineering, 1(2), pp. 17-23, 2014
21
of the antibacterial activity using the parameters
proposed by Alves et al. (2000) where zones of
inhibition are rated as: < 9 mm (inactive); 9 - 12 mm
(less active); 13 - 18 mm (active) and > 18 mm (very
active) that the extracts that even showed a weak
activity are inactive. In comparison with report by
Doughari (2010) this result showed that the plant does
not have a broad spectrum of activity. This narrow
spectrum of activity may be suggestive of the fact that
there is a synergistc effect of the phytoconstituents of
the extract obtained by extracting the plant material
with a polar solvent instead of successive extraction
with solvents of varying polarity.

5. CONCLUSION

The result of this study clearly revealed that the
antioxidant activity of the root bark of coral tree
compares favourably with that of the reference
(Vitamin C) used and it holds promise for future
application as a natural substitute to the hitherto
available standard but expensive antioxidants. This
finding is one too significant to be ignored and
evidence previously unobtainable being the first report
on the antioxidant activity of the root bark of the
plant. However, toxicological investigations for
ascertaining the clinical safety and therapeutic dose of
the plant extracts should be carried out.

ACKNOWLEDGEMENTS

The authors acknowledge Messrs Elkanah Daniel and
Salihu E. Yunusa for their assistance in the study.

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International Journal of Scientific Research in Chemical Engineering, 1(2), pp. 17-23, 2014
23




Adeniran Oluremi Isola Lectures at the Department of Chemistry, Faculty of Science, University of
Abuja, Nigeria. Adeniran holds an M. Sc. Degree in Organic Chemistry from the University of
Ibadan. He has a burgeoning research prospect in Natural Products/Medicinal Plants and has
published some scientific articles in the field. He has also served in the capacity of Course
Writer/Developer in the course team for Natural Products Chemistry of a certain institution.










ABIMBADE, Sunday Felix obtained his B.Tech degree in Pure and Applied Chemistry from Ladoke
Akintola University of Technology (LAUTECH), Nigeria in 2006. He also holds an M.Sc. degree in
Organic Chemistry from the University of Ibadan. Currently, he is running his PhD programme in the
University of Ibadan and he is a Lecturer in Federal University, Oye Ekiti. He has to his credit some
published scientific articles in reputable local and international journals. His research focus has been
on Natural Products Chemistry but recently, he added synthesis of medicinal natural products to his
research career.