Pharmacogn J.

2021; 13(6): 1355 -1358

A Multifaceted Journal in the field of Natural Products and Pharmacognosy
Original Article
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Phytochemical Composition and Antibacterial Activities of

Syzygium polyanthum Methanolic Leaves Extract
Noor Zarina Abd Wahab1,*, Nur Saidatul Aqilah Ja’afar2

ABSTRACT
Background: Since a long time ago, Syzygium polyanthum has become traditional herb for health, and thus
many studies done to confirm the medical effectiveness. Objectives: This present study was conducted
to evaluate phytochemical compounds and antibacterial activities of methanolic extract of S. polyanthum
leaves. Materials and Methods: In the present study, standard methods of determination were used to
Noor Zarina Abd Wahab1,*, Nur determine the phytochemical compounds of S. polyanthum. The methanol extract of S. polyanthum leaves
Saidatul Aqilah Ja’afar2 were subjected for antimicrobial activity. Antibacterial activity was evaluated against five bacterial strains
by determining minimum inhibitory concentration (MIC) and zone of inhibition. Diameters of the zone of
1
Department of Biomedicine, Faculty of inhibition were compared with standard antibiotics. Results: Phytochemical analyses qualitatively reveal
Health Sciences, Universiti Sultan Zainal the presence of alkaloids, saponin, terpenoids and steroid. The extract was found to exert antibacterial
Abidin, Terengganu, MALAYSIA. activity against four tested bacteria which are S. aureus, S. pyogenes, MRSA and K. pneumoniae except
2
School of Health Sciences, Universiti Sains
for E. coli. The zones of inhibition shown by the disc diffusion method for S. aureus were between 8.6
Malaysia Health Campus, Kubang Kerian,
Kelantan, MALAYSIA. to14.0 mm, S. pyogenes were between 8.4 to 12.0 mm, MRSA was between 10.0 to 13.2 mm and K.
pneumoniae were between 8.0 to 10.6 mm. Meanwhile, there was no zone of inhibition was observed
Correspondence for E. coli. The MIC was determined ranging from 6.25 to 12.5 mg/ml against all the tested bacteria.
Noor Zarina Abd Wahab The highest value of MIC showed by S. aureus, S. pyogenes, MRSA and K. pneumoniae which is 6.25
Department of Biomedicine, Faculty of
mg/mL. Conclusion: The tested leaves extract showed promising antibacterial activity against both
Health Sciences, Universiti Sultan Zainal Gram positive and Gram-negative bacteria. Phytochemical screening revealed the presence of alkaloids,
Abidin, Terengganu, MALAYSIA. saponin, terpenoids and steroid in methanolic leaves extract qualitatively and these compounds could be
E-mail: [email protected] responsible for antibacterial properties of leaves extract of S. polyanthum.
Key words: Syzygium polyanthum; Phytochemical; Antibacterial.
History
• Submission Date: 14-07-2021;
• Review completed: 26-07-2021; INTRODUCTION which is responsible for the antibacterial activity of
• Accepted Date: 23-08-2021. the extract. Both extracts were observed to inhibit
Antibacterial resistance is a serious clinical Gram positive and Gram negative bacteria tested in
DOI : 10.5530/pj.2021.13.171
challenge worldwide because some of the bacteria vitro to suggest that S. polyanthum leaves extract and
Article Available online develop enzymes that are capable of digesting fractions has a broad-spectrum antibacterial activity.
http://www.phcogj.com/v13/i6 and destroying antibiotic molecules. Therefore,
Copyright this emerging issue has urged the scientist to Previous studies have discussed the phytochemical
© 2021 Phcogj.Com. This is an open- find a potential source of treatment to encounter constituents of S. polyanthum especially that focused
access article distributed under the terms that problem. There are numerous reports on on the plant leaves. Gas Chromatography-Mass
of the Creative Commons Attribution 4.0 the inhibitory effects of various plant extracts Spectrometry (GC-MS) analyses organic crude ex-
International license.
on the growth of many bacteria in culture.1-3 tracts from S. polyanthum leaves on (hexane, ethyl
Syzygium polyanthum (Wight) Walp ” which is acetate and methanol) showed that each crude ex-
also recognized as “salam”, “serai kayu” or “samak tract was composed of different phytochemical com-
kelat” by Malay people is a tropical plant.4 S. positions.7 Overall, the study conducted revealed
polyanthum leaves, fruits and barks have been used that the crude extracts composed of hydrocarbons,
traditionally as medicine or therapeutic agents aldehydes, terpenoids, phenolics, fatty acids, mono-
including efficiency against ulcer, hypertension, terpenes, diterpenes, triterpenes and sesquiterpenes.
diabetes mellitus, hyperuricemia, diarrhea, There are nine compounds present in all three ex-
gastritis, skin diseases and inflammation.5 tracts include nerolidol, caryophyllene oxide, farne-
Furthermore, various bacteria and fungi have sol, phytol, squalene, β- tocopherol, γ-tocopherol,
been tested by extracts from leaves, fruits, as well α-tocopherol and β-sitosterol. Cytotoxic compounds
as essential oil of S. polyanthum. Plant extracts such as linalool and octanal, α-humulene, and pyro-
are studied and found to have phytochemical gallol (1,2,3-benzenetriol) also have been discovered
constituents such as phenolic compounds, in that study. Thus, this study, was carried out to
protein-like compounds, flavonoids, and tannins evaluate phytochemical constituent and antibacte-
that possess antimicrobial activity. Extracts rial activity of S. polyanthum methanol leaves extract
and fractions of S. polyanthum leaves had a low in search for active antimicrobial activity.
inhibition and/or can inhibit the bacteria P.
aeruginosa and B. subtilis. The highest antibacterial MATERIALS AND METHODS
activity against bacteria S. aureus and E. coli is
shown using the n-hexane extracts.6 This can be
Plant materials and extraction
explained by the active compounds in the non- The fresh plant of S. polyanthum was collected
polar fractions in relatively higher concentration from the state of Terengganu, Malaysia. The plants

Cite this article: Abd Wahab NZ, Jaafar NSA. Phytochemical Composition and Antibacterial

Phcogj.com
Activities of Syzygium polyanthum Methanolic Leaves Extract. Pharmacogn J. 2021;13(6):
1355 -1358.

1355 Pharmacognosy Journal, Vol 13, Issue 6, Nov-Dec, 2021

 Abd Wahab NZ, et al.: Phytochemical Composition and Antibacterial Activities of Syzygium polyanthum Methanolic Leaves Extract

authentication was performed by competent botanist from Faculty sets of standardized microtiter plates with shallow wells that contain
of Bioresources and Food Industry, Universiti Sultan Zainal Abidin. increasing dilutions (decreasing concentration) of plant extract was
The plants were cleaned with tap water to remove dirt and oven-dried inoculated with bacterial suspensions. The plates were incubated
at 60°C before homogenized into a fine powder. Leaves powder was at 37 °C for 24 h. The lowest concentration of the extract in the well
weighed into 200 g then was immersed in methanol (1500 mL) for of the microtiter plate that showed no turbidity after incubation are
three days at room temperature. After that, the mixture was filtered MIC. Bacterial suspensions were used as negative control, while broth
to make sure that all the mixture was filtered completely. The filtrate containing standard drug was used as positive control.
was concentrated in a rotary evaporator (Eyela N-1110, Japan) under
a controlled vacuum. The extract was stored in the freezer till the use.8 RESULTS
Phytochemical screening Phytochemical analysis
Phytochemical qualitative screening was carried out for S. polyanthum Phytochemical analysis shown in Table 1 indicates the methanolic
extract according to standard procedures used to test for the presence of extract of S. polyanthum leaves contained a few secondary metabolites
alkaloids, saponins, terpenoid and steroids.9 such as alkaloids, saponin, terpenoids and steroid.

Determination of alkaloid Disk diffusion test

10 mg/mL of extract was diluted in 2 mL of 25% ammonia for several The inhibition zone of S. polyanthum methanolic leaves extract against
minutes. After that, 5 mL of chloroform was added and shaken gently to Gram positive and Gram negative bacteria is shown in Table 2. Results
extract the alkaloidal base. Mayer’s reagent was added. The formation obtained in the present study revealed that the methanolic extract of
of a cream with Mayer’s reagent was observed indicating the presence S. polyanthum leaves exhibits antibacterial activity against S. aureus,
of alkaloids. MRSA, S. pyogenes and K. pneumoniae but not E. coli. The inhibition
zones were between 8.0 and 14.0 mm. The larger inhibition zone gave
Determination of saponin the meaning of higher antibacterial activity of the extract on the tested
microbial species.
2 mL of distilled water was added to extract suspended in ethanol and
was shaken vigorously. The formation of copious foam layer indicates Determination of Minimum Inhibitory Concentration
the presence of saponins.
(MIC)
Determination of terpenoid From the result shown in Table 3, S. polyanthum methanolic leaves
2 mL of chloroform was added to 10 mg of the extract. After that, 3 mL extract demonstrated broad-spectrum activity against four selected
of concentrated sulphuric acid (H2SO4) was carefully added to form
a layer. A reddish brown colouration of the interface indicates the Table 1: Phytochemical analysis of S. polyanthum methanolic leaves
presence of terpenoids. extract.
Phytochemical substances +: Present; -: Absent
Determination of steroid Alkaloid +
2 mL of chloroform and 2 mL of concentrated H2SO4 was added to Saponin +
10 mg/mL of extract and shaken well. Chloroform layer appeared red. Terpenoids +
This confirm the presence of sterols steroid. Steroid +

Bacteria cultures Table 2: Zone of inhibition for disc diffusion method of S. polyanthum
Two pathogenic Gram positive bacteria, Staphylococcus aureus (ATCC methanolic leaves extract against selected bacteria.
11632), clinical isolate methicillin-resistant Staphylococcus aureus Zone of inhibition (mm)
Concentration of
(MRSA) and three pathogenic Gram negative bacteria Streptococcus
extract (mg/mL) S. aureus S. pyogenes MRSA E. coli K. pneumoniae
pyogenes (ATCC 12344), Klebsiella pneumoniae (ATCC 10031) and
Escherichia coli (ATCC 10536) were used. Bacterial strains were grown 125 8.6 8.4 10.0 - 8.0
and maintained on Mueller-Hinton agar (MHA). 250 10.6 10.0 12.0 - 8.0
Disk diffusion test 500 14.0 12.0 13.2 - 10.6

S. polyanthum extract was tested for antimicrobial activity using the 10% methanol
- - - - -
(negative control)
disk diffusion method. Overnight cultures (107-109 CFU mL-1) of
bacteria strains were spread on Mueller Hinton agar (MHA) with Chloramphenicol
28.0 30.0 22.0 24.0 26.0
(positive control)
a sterile cotton swab. Sterile filter paper discs with 6 mm diameter
were placed on top of the culture and 20 µL of 125 mg/mL, 250 mg/ -: No inhibition activity
mL and 500 mg/mL (w/v) of S. polyanthum extract were loaded on the Zone of inhibition (mm) are average of triplicate experiments. Disc diameter = 6 mm.
paper discs. The 10% methanol-loaded disc was used as the negative
Table 3: MIC value of S. polyanthum methanolic leaves extract.
control. All impregnated discs were fully dried at room temperature
in a laminar flow hood before the application on the bacterial lawn. Bacterial Strains Tested MIC (mg/mL)
Antibacterial activity was evaluated by measuring the diameter of the S. aureus 6.25
inhibition zone (in mm) produce by the leaf extract around the discs.10 S. pyogenes 6.25
MRSA 6.25
Determination of minimum inhibitory concentrations (MIC) K. pneumoniae 6.25
MIC was performed to determine the lowest concentration of the E. coli 12.5
plant extract that inhibits the growth of a microorganism.11 Two The MIC values (mg/mL) are average of triplicate experiments.

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 Abd Wahab NZ, et al.: Phytochemical Composition and Antibacterial Activities of Syzygium polyanthum Methanolic Leaves Extract

bacteria with the MIC values ranging from 6.25 to 12.5 mg/mL. Results ACKNOWLEDGEMENTS
showed that all of the tested bacteria were susceptible pathogens with the
MIC value of 6.25 mg/mL except E. coli with MIC value of 12.5 mg/mL The authors would like to thank Universiti Sultan Zainal Abidin
(UniSZA) for the facilities and laboratory instruments.
DISCUSSION
CONFLICTS OF INTEREST
Medicinal plants have long been deployed as the basis of treatment
of various diseases in traditional medicine such as in Asian and also None.
as worldwide ethnomedicine.12,13 The ethnomedicinal plant is one of
the important sources for drug discovery and Syzygium polyanthum
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GRAPHICAL ABSTRACT

ABOUT AUTHORS

Dr. Noor Zarina Abd Wahab is senior lecturer, researcher at the Department of Biomedicine,
Faculty of Health Sciences, Universiti Sultan Zainal Abidin, Malaysia. Her current research
interest is antimicrobial mechanism of action and biological activity of plant natural products.

Nur Saidatul Aqilah Ja'afar is medical laboratory scientist at the School of Health Sciences,
Universiti Sains Malaysia. Her research interests is in medical microbiology.

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