Biological
and Clinical Sciences Research Journal
ISSN: 2708-2261
www.bcsrj.com
DOI: https://doi.org/10.47264/bcsrj0201015
Biol. Clin.
Sci. Res. J., Volume, 2021: e015
Original Research Article
ANTIOXIDANT
ACTIVITY OF SYZYGIUM AROMATIUM AND CINNAMOMUM VERUM SEED EXTRACTS
MAHMOOD H, ALI Q, *HAFEEZ MM, MALIK A
Institute of
Molecular Biology and Biotechnology, The University of
Lahore, Lahore, Pakistan
Corresponding author: mansoorhafeez140@gmail.com
Abstract
The
present study was carried out for the evaluation of antioxidant activities of
clover (Syzygium aromatium L.) and
cinnamon (Cinnamomum verum
L.). The research work was carried out at the Institute of Molecular Biology,
University of Lahore. The samples of clovers and
cinnamon were collected from Lahore and were crushed to prepared sample in n-hexan, and ethanol extracts for evaluation of antioxidant
activities. The highest antioxidant activities of ethanol extract from cinnamon
indicated that the cinnamon may be used as an active ingredient to control
stress conditions of cells for stress avoidance. The alkaloids, coumarins, saponins, flavonoids, tannins, sterols, quinons,
triterpenoids, anthocyans, leucoanthocyans and terpenoids
were tested for both ethanol and n-hexan extracts
which indicated that most of the components were found present in both of the
plant species, which revealed that these herbal plant seeds may be used as
potential medicinal plants.
Keywords:
clovers,
cinnamon, antioxidant, ethanol, n-hexan
Introduction
As shown from the chemical diversity, the saponins have a very wide range for biological activities
collectively. Many saponins have antioxidant,
antibacterial and antiherbivore activity so these
play a vital role in plant defense mechanism (Lo et al., 2004; Roby et al.,
2013). The use of cinnamon (Cinnamomum verum L.) also showed antioxidant and antimicrobial
activities against different bacterial strains including Bacteroides succinogenes, E. coli, Streptococcus bovis, Staphylococcus aureus, Ruminococcus flavefaciens and
fungal strains including Penicillium roqueforti, Mucor plumbeus, Zygosaccharomyces rouxii, Candida
lipolytica, Debaryomyces hansenii, Aspergillus flavus, Pediococcus halophilus and Pichia membranaefaciens. The use of cinnamon oil was carried
out for the inhibition of microbial growth under control conditions. The
cinnamon (Cinnamomum verum L.)
largely produced by Indonesia up to 43% among all of the cinnamon producing
countries while China contributes up to 33% in total world production of
cinnamon (Diao et
al., 2014; Zhang et al., 2016).
The clovers (Syzygium aromatium L.)
also an important plant which shows antioxidant and antimicrobial activities
against bacterial strains like Clostridium
E. coli, sticklandii, salmonella along with at some extant
against fungal strains. The antioxidant and antimicrobial activities of clovers
are due to the presence of soluble phenolics. The
leaves of clovers are usually rich in with isoflavonoids,
biochanin A and formioninetin
(Anwar et al., 2009; Singh and Majumdar, 1999). The presence of pehnolic
compounds showed the inhibitory activities against the growth and multiplication
of microbes. The phenolic compounds in clovers also
caused the fermentation of amino acids. The clover leaves and shoots may be
used as antioxidants and antimicrobial medicines to prevent human, food and
animals from attacks of microbes (Chesson et al., 1982; Dickinson et al., 1988; Lee et al., 2004). The chemical contents have been used for the
formation of fungicidal lotions, antioxidant, pesticides and insecticides due
to its insect repellent activities (Dua et al., 2013; Kaithwas
et al., 2011; Singh et al., 2005). The sterols and aglycone triterpenes themselves
encompass a wide range of structure with the nature of cyclization,
oxidation in structure and variation in the degree. Secondly, variation with
respect to sugar molecules and their numbers and nature of glycosylation,
the presence of sugar chains and types of inter sugar chain linkages. Single
sugar is attached at C-3 position in mono desmosidic saponins. While at C-28 position in bidesmosidic
saponins (like triterpenoid
saponins), and in steroid saponins
sugar are attached at C-26 position (Chesson and
Forsberg 1997; Sullivan, 2009). The rumen bacterial strains caused disease in
the intestine of animals which may lead towards the death of animal. These
bacterial strains can be controlled or killed through the use of plant phytochemical or phenolic
compounds as antimicrobial agents which are also used as potential antioxidants
(Chesson et al.,
1982; Stewart et al., 1997).
Materials and
methods
The research was conducted in Microbiology
Lab at the Institute of Molecular Biology and Biotechnology, The University of
Lahore. The research work was done in the time period from November 2019 to
March 2020. The
seed samples were included Syzygium aromatium and Cinnamomum verum (Trifolium pretense L.) which was collected from market of Lahore, Pakistan.
Preparation
of plant extracts
The dried samples were brought to the laboratory and
crushed into small pieces by using laboratory
grinder. After grinding for clovers (Syzygium aromatium and Cinnamomum verum (Trifolium pretense L.) were dipped in two different solvents ethanol and n-hexan ether depending on the amount required and kept for 2 weeks with vigorous shaking but the amount of
samples was kept the different. All
samples were taken in grams (g) and solvents per milliliters (ml). After keeping the samples in the solvents for 2 weeks and
samples were filtered using a whatman filter paper
into a new bottle and then placed in the rotary evaporator. After the rotary process, samples were collected in falcon
tubes and labeled accordingly and then placed
in Petri plate for at least 1-2 days to evaporate completely. Dried extracts
were collected and stored at refrigerator 4°C for further used
detail shown in table below.
Antioxidant
activity
Antioxidants are capable to inhibit or
either delayed oxidation processes, which occurred
under the effects of the atmospheric O2 or ROS (reactive oxygen
species). They were used to stabilize polymeric products of
foodstuffs, pharmaceuticals,
petrochemicals and cosmetics. Antioxidants are involved in
defense mechanism or immune system of
organism against varieties of pathologies which are usually associated with the
attacked of free oxygen radicals or ROS. To check the antioxidant
properties of clovers
(Syzygium aromatium L.) and cinnamon
(Cinnamomum verum L.) plant DPPH assay was applied:
2, 2-Diphenyl-1-picrylhydrazyl (DPPH) assay
DPPH, known formally as 2, 2-dipheny1-1-picrylhydrazl, was a
cell-permeable, stable
free radical that was commonly used to evaluate the ability of compounds to
acted as free radical scavengers or hydrogen
donors and to measure the antioxidant activity of flower, leaves, stem and roots extracts. The reaction of DPPH with an
antioxidant or reducing compound
produced the corresponding hydrazine DPPH, which could be followed by
color changed from purple (absorbance at 515-528nm) to yellow (Adedayo et al., 2004). The
antioxidant activity of the extracts was determined using the DPPH free radical
scavenging assay described with some modifications. Briefly, the
universal bottle was contained 501.1L of clovers
(Trifolium pretense L.), Alsi (Linum usitatissimum L.), fennel or sonf (Foeniculum vulgare L.), cinnamon (Cinnamomum verum L.) extracts in concentrations from 1 to 5
mg/n11, 0.004% (w/v) solution of DPPH was
added. The obtained mixture was vortexed, incubated
for 30 min in room temperature in a relatively dark place and then was read
using spectrophotometer at 517 nm. The blank was 80% (v/v) methanol. Ascorbic
acid (vitamin C) was used for comparison. Measurements were taken in
triplicate. DPPH scavenging effect was calculated using the following equation:
DPPH scavenging effect (%) = [(A0 – A1)/A0)
× 100]
Where AO was the absorbance of negative control (0.004%
DPPH solution) and A was the absorbance in presence of extract. The results were
reported as IC50 values and ascorbic acid equivalents (AAE, mg/g) of clovers (Syzygium aromatium L.) and cinnamon
(Cinnamomum verum L.) extracts.
Results and discussions
The
ethanol and n-hexan extracts from clover and cinnamon were used to find out the antioxidant
activities present in the spices plants. The highest antioxidant activity
(2.678) through ethanol plant extract at 50 µg/ml concentration was reported by
cinnamon sample while lowest (0.267) was recorded for clover. The highest antioxidant activity (0.568)
through ethanol plant extract at 100 µg/ml concentration was reported by cinnamon
sample while lowest (0.557) was recorded for clover. The highest antioxidant activity (2.80,
1.173, 300) through ethanol plant extract at 200 µg/ml, 150 µg/ml and 250 µg/ml
concentrations was reported by Cinnamon sample while lowest (0.228, 0.153,
0.238) was recorded for clover respectively
(Table 1). The lowest antioxidant activity (0.231) through n-hexan plant extract at 50 µg/ml concentration was reported
by clover sample while
higher (0.275) was recorded for Cinnamon. The highest antioxidant activity
(0.714) through n-hexan plant extract at 100 µg/ml
concentration was reported by cinnamon sample while lowest (0.411) was recorded
for clover. The highest
antioxidant activity (1.241) through n-hexan plant
extract at 150 µg/ml concentration was reported by cinnamon sample while lowest
(0.171) was recorded for clover.
The highest antioxidant activity (1.752) through n-hexan
plant extract at 250 µg/ml concentration was reported by cinnamon sample while
lowest (0.464) was recorded for clover.
The highest antioxidant activity (0.565) through n-hexan
plant extract at 200 µg/ml concentration was reported by cinnamon sample while
lowest (0.232) was recorded for clover
(Table 2). The alkaloids, coumarins, saponins, flavonoids, tannins,
sterols, quinons, triterpenoids,
anthocyans, leucoanthocyans
and terpenoids were tested (Tables 3) for both
ethanol and n-hexan extracts of clover and cinnamon
which indicated that most of the components were found present in both of the
species, which revealed that these herbal plants may be used as potential
medicinal plants. The ethanol and n-hexan extracts
from clovers and
cinnamon were used to find out the antioxidant activities present in the spices
plants. The highest antioxidant activities of ethanol extract from Cinnamon and
clover indicated that the cinnamon and clover may be used as an active
ingredient to control stress conditions of cells for stress avoidance. The
highest antioxidant activities of ethanol extract from Cinnamon indicated that
the cinnamon may be used as an active ingredient to control stress conditions
of cells for stress avoidance. The highest antioxidant activities of ethanol
extract from Cinnamon and clover indicated that the cinnamon and clover may be
used as an active ingredient to control stress conditions of cells for stress
avoidance (Anwar et al., 2009; Heng
et al., 2006; Lee et al., 2004; Matan
et al., 2006).
Table
1. Antioxidant
activities of spices by ethanol extract
|
Doses |
clover (Syzygium aromatium L.) |
Cinnamon (Cinnamomum verum L.) |
|
50 µg/ml |
0.267 |
2.678 |
|
100 µg/ml |
0.557 |
0.568 |
|
150 µg/ml |
0.228 |
2.80 |
|
200 µg/ml |
0.153 |
1.173 |
|
250 µg/ml |
0.238 |
3.00 |
Table 2. Antioxidant activities of spices by n-hexan extract
|
Doses |
clover (Syzygium aromatium L.) |
Cinnamon (Cinnamomum verum L.) |
|
50 µg/ml |
0.231 |
0.262 |
|
100 µg/ml |
0.411 |
0.714 |
|
150 µg/ml |
0.171 |
1.241 |
|
200 µg/ml |
0.232 |
0.565 |
|
250 µg/ml |
0.464 |
1.752 |
Table 3. Phytochemical composition of clover (Syzygium aromatium L.) and Cinnamon (Cinnamomum verum L.) seed extract
|
|
clover (Syzygium |
aromatium L.) |
Cinnamon (Cinnamomum |
verum L.) |
|
Phytochemical |
Ethanol extract |
n-hexan
extract |
Ethanol extract |
n-hexan extract |
|
Alkaloids |
+ |
+ |
+ |
+ |
|
Coumarins |
+ |
+ |
+ |
+ |
|
Saponins |
- |
+ |
+ |
+ |
|
Flavonoids |
+ |
+ |
- |
+ |
|
Tannins
|
+ |
- |
- |
- |
|
Sterols |
+ |
+ |
+ |
+ |
|
Quinons |
+ |
+ |
+ |
+ |
|
Triterpenoids |
- |
- |
+ |
- |
|
Anthocyans |
- |
- |
+ |
- |
|
Leucoanthocyans |
+ |
+ |
- |
+ |
|
Terpenoids |
+ |
+ |
+ |
+ |
Conclusions
The alkaloids, coumarins, saponins, flavonoids, tannins, sterols, quinons,
triterpenoids, anthocyans, leucoanthocyans and terpenoids
were tested for both ethanol and n-hexan extracts of clover (Syzygium aromatium L.) and Cinnamon
(Cinnamomum verum L.)
which indicated that most of the components were found present in both of the
species, which revealed that these herbal plants may be used as potential
medicinal plants. It was concluded that the Cinnamon (Cinnamomum verum L.) and clover (Syzygium aromatium L.) may be used as potential antibiotics
and antioxidants.
Conflict of interest
The
authors declared absence of conflict of interest.
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