ANTIBACTERIAL EFFICIENCY OF POMELO PEEL EXTRACT ON VARIOUS

CONCENTRATIONS AGAINST SELECTED MICROORGANISMS

Anna Cariza Ayad, Karen Nicole Buerano, Sony Andrew Siregar Sormin,
Carmela Malabat, Susy Jael, Lea Divina, Jo-Anne Lucero

Abstract

T
his study determined the phytochemical components and antibacterial efficiency of Citrus maxima (pomelo)
peel extract on two concentrations, 75% and 95%, against selected microorganisms. The phytochemical
analysis revealed the presence of alkaloids, flavonoid, glycosides, saponins, sterols, tannins, and triterpines.
Alkaloids were abundantly found while only traces of the other constituents were found. E. coli and P. aeruginosa
both produced 10mm complete inhibition in 75% and 95% extract concentrations. On the other hand, S. aureus
produced slight inhibitory activity with a mean zone of inhibition of 10mm against the 75% extract concentration.
It also produced partial inhibitory activity with a mean zone of inhibition of 10mm against 95% concentration. In
comparison, the antibiotic Levofloxacin which served as appositive control for E. coli and P.aeruginosa produced
17.92mm and 16.85mm complete inhibition for the 75% extract concentration. On the 95% extract concentration,
Levofloxacin produced 18.73mm and 18.70mm complete inhibition. For the positive control of S. aureus,
Clindamycin was utilized, which produced 16.30mm complete inhibition in the 75% concentration and 15.02mm
complete inhibition in the 95% extract concentration. The results showed that pomelo peel extract is effective in
inhibiting the growth of bacteria, and the difference in concentrations was significant for S. aureus.

Keywords: phytochemical, antibacterial efficiency, Citrus maxima (pomelo) peel extract

Ever since the birth of mankind, there has been a relationship against several bacteria such as Staphylococcus aureus, Bacillus
between life, disease, and plants. Citrus maxima (pomelo), also subtilis, Bacillus cereus, Salmonella enteritidis, and Escherich-
known as Chinese grapefruit, belongs to the rue family (Ruta- ia coli.
ceae) and is the largest citrus fruit. It is native to the Southeast The human skin normally serves as a primary line of
Asia and Indo-China regions (Cheong, Shao, Zhou, & Yu, 2012) defense against infections caused by the aforementioned mi-
and is one of the most important horticultural crops growing croorganisms. However, some bacteria are found in normal flo-
extensively in tropical and subtropical southern regions of Asia ra which can turn opportunistic once there is breakage on the
(Lan-Phi & Vy, 2015). skin. Normal flora found in the skin includes Staphylococcus
The pomelo tree is a perennial shrub that grows 16- aureus, which is mostly localized in the nose and other orofa-
50 feet (5-15 meters) tall. Its peel may be greenish-yellow or cial areas (Tognetti, Martinelli, Berti, Hercogova, Lotti, Leon-
pale-yellow while the pulp varies from greenish-yellow or cini, & Moretti, 2012). Pseudomonas aeruginosa could cause
pale-yellow to pink or red. The fruit’s taste varies from mildly life-threatening infections in people who have compromised
sweet and bland or rather acidic, with a faint touch of bitterness immune systems in different areas of the body such as the skin
(Cheong, Shao, Zhou, & Yu, 2012). Guo and Abeysinghe as cit- (Koehnke & Friedrich 2015). Escherichia coli strains are fre-
ed in Toh, Khoo, and Azrina, (2013) state that the peel of the cit- quently isolated from skin and soft tissue infections (Petkovsek,
rus fruit contains a higher amount of antioxidants as compared 2009). It is a major cause of diarrheal diseases, peritonitis, coli-
to its pulp, as the purpose of the peel is to protect the antioxi- tis, bacteremia, infant mortality, and urinary tract infections
dants in the fruit from oxidation. Therefore, it is recommended (Blount, 2015).
to consume the fruit together with its peel. Commonly used as a sweet delicacy, pomelo is over-
The pomelo fruit has been linked to several purposes. It looked as a source of prevention in microorganism proliferation.
is rich in vitamin C and has been used in indigenous medicine as A study by Muhammad (2015) showed that citrus fruits exhibit-
a sedative for nervous affections and convulsive cough, and as ed antibacterial activity against Escherichia coli, Pseudomonas
a treatment for hemorrhagic disease and epilepsy (Vijaylakshmi aeruginosa and Staphylococcus aureus. Further, the effect of
& Radha, 2015). According to Arias and Ramon-Laca as cited in crude extract from pomelo peel can be used to inhibit Staphylo-
Caengprasath, Sathaporn, Kittana, and Siricahi (2012), the pulp coccus aureus (Aichayawanich, 2012).
of the pomelo was used as an appetizer, antitoxic, cardiac stim- In view of previous findings regarding the high antimi-
ulant, and stomach tonic. Furthermore, Mokbel and Suganuma crobial and anti-oxidant activities of a number of phytochemi-
as cited in Naradisorn and Ruenkum (2009) state that pomelo cals inherent in citrus fruits, the pomelo fruit can therefore be
extracts have been revealed to have an antimicrobial activity a potential replacement for synthetic preservatives as all citrus
76 Journal of Health Sciences

fruits have similar complex structures regardless of cultivars. Therefore, the pomelo holds potential for providing multiple benefits
to consumers by way of its possible usage in the fields of medicine, therapeutics, and food technology (Barrion, Hurtada, Papa,
Zulayvar, & Yee, 2014). Due to the components found in the pomelo peel, the antibacterial properties may aid in eliminating the
bacteria found in a person’s skin. This study was conceptualized to explore this potential and provide evidence of the fruit’s antibac-
terial efficiency.

Purpose of the Study

This study determined the phytochemical components and antibacterial efficiency of Citrus maxima (pomelo) peel extract
on two concentrations, 75% and 95%, against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus.

METHODOLOGY
This study utilized an experimental research design. All the microorganisms were preserved and obtained from the Depart-
ment of Science and Technology. The extract underwent phytochemical analysis and antimicrobial activity test using the disc agar
diffusion method or Kirby-Bauer Test on the three selected bacteria.

Collection and Preparation of Pomelo Peels

Pomelo fruit peels were collected and placed in a clean air-tight container. The peels were thinly sliced for increased dry-
ing of moisture content. After slicing, the peels were placed in a container for weighing. Around 500 grams of pomelo peel were
weighed. For drying, the peels were placed inside the Multi Commodity Heat Pump Dryer overnight. The peels were weighed again
after an hour of drying to ensure the decrease in moisture content. The peels were placed in a blender and grounded until it turned
into powder. They were afterwards placed in a clean plastic container.

Preparation of Pomelo Peel Extract

One hundred grams of the ground plant material were weighed in an Erlenmeyer flask. Three hundred milliliters of 80%
ethyl alcohol were then added to completely submerge the materials. The solution was stoppered and soaked for 24 hours. It was
filtered through a Buchner funnel using gentle suction. The flask and the plant material with fresh portions of alcohol were rinsed.
The washings with the first filtrate were combined and the plant residue was discarded.
The filtrate under vacuum was concentrated to a syrupy consistency or about 20 milliliters. The exact volume of the con-
centrated extract was measured. This is the strength of the extract expressed in grams of plant material per milliliter of the extract.
The extract was stored in a tightly stoppered container in a cold environment (0-5°C). The extract was then considered ready for
phytochemical and microbiological screening. The extract was weighed for quantitative determinations in biological tests.
For fresh plant material, 200 grams of the finely cut fresh material were used and soaked in 300 milliliters of 95% ethyl
alcohol to completely submerge the material.

Phytochemical Analysis of the Pomelo Peel Extract

Following the extraction, the pomelo peels underwent phytochemical analysis to test its chemical constituents. All the
chemicals used for testing were purchased from the Department of Science and Technology and the process of analysis was based
on the procedures of the Standards of Testing Division where the test was conducted.

Disc Agar Diffusion Method or Kirby–Bauer Test

Kirby-Bauer test was performed under the standard conditions of the facility. The main purpose of this test was to calculate
the inhibitory concentration for a given antibiotic by comparing the observed zone of inhibition’s size to known values.

RESULTS AND DISCUSSION

Antimicrobial Activity Test Result
Table 1 presents the antimicrobial activity test results of the pomelo (Citrus maxima) peel extract of 75% concentration
against three microorganisms.

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 Antibacterial Efficiency of Pomelo Peel Extract on Various Concentrations Against Selected Microorganisms 77

Table 1
Antimicrobial ActivityTest: 75% Concentration of Ethanol Extract
Escherichia coli Pseudomonas aeruginosa Staphylococcus aureus
Sample/Control Total Reactivity Inhibitory Total Reactivity Inhibitory Total Reactivity Inhibitory
Mean Activity Mean Activity Mean Activity
Zone of Zone of Zone of
Inhibition Inhibition Inhibition
Pomelo Peel Ethanolic Extract 10 2 +++ 10 2 +++ 10 2 +
75% (10mm)
Levofloxacin 5 ug (6-mm 17.92 4 +++ 16.85 4 +++ N/A N/A N/A
positive control)
Clindamycin 2 ug (6-mm N/A N/A N/A N/A N/A N/A 16.3 4 +++
positive control)

The 75% ethanol extract showed that Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus are reactive
organisms to the pomelo peel extract. The bacteria Escherichia coli produced complete inhibitory activity (+++) with a total mean
zone of inhibition of 10 millimeters and had a mild reactivity rating (2) from the extract. This is in contrast with one study by Bar-
rion, Hurtada, Papa, Zulayvar, and Yee (2014) where E. coli was observed to be resistant to all the sample extracts at all concentra-
tions. The findings also show that Pseudomonas aeruginosa produced complete inhibitory activity (+++) with a total mean zone of
inhibition of 10 millimeters and had mild reactivity (2) against the pomelo peel.
In comparison, the antibiotic Levofloxacin (5 ug) which served as a positive control for Escherichia coli and Pseudomonas
aeruginosa produced complete inhibitory activity (+++) with a total mean zone of inhibition of 17.92 and 16.85 for for Escherichia
coli and Pseudomonas aeruginosa, respectively. Both resulted to a severe reactivity rating (4).
On the other hand, Staphylococcus aureus produced slight inhibitory activity (+) with a mean zone of inhibition of 10
millimeters and had mild reactivity (2) against the extract concentration. For the positive control of the Staphylococcus aureus,
a different antibiotic, Clindamycin (2 ug), was utilized. It produced complete inhibitory activity (+++) with a total mean zone of
inhibition of 16.30 and a severe reactivity rating (4) for Staphylococcus aureus.

Table 2
Antimicrobial Activity Test: 95% Concentration of Ethanol Extract
Escherichia coli Pseudomonas aeruginosa Staphylococcus aureus
Sample/Control Total Reactivity Inhibitory Total Reactivity Inhibitory Total Reactivity Inhibitory
Mean Activity Mean Activity Mean Activity
Zone of Zone of Zone of
Inhibition Inhibition Inhibition
Pomelo Peel Ethanolic Ex- 10 2 +++ 10 2 +++ 10 2 ++
tract 95% (10mm)
Levofloxacin 5 ug (6-mm 18.73 4 +++ 18.7 4 +++ N/A N/A N/A
positive control)
Clindamycin 2 ug (6-mm N/A N/A N/A N/A N/A N/A 15.02 3 +++
positive control)

Table 2 shows the antimicrobial activity test results of the pomelo (Citrus maxima) peel extract of 95% concentration
against three microorganisms. The 95% ethanol extract showed that Escherichia coli, Pseudomonas aeruginosa, Staphylococcus
aureus are reactive organisms to the pomelo peel.
Escherichia coli produced complete inhibitory activity (+++) with a total mean zone of inhibition of 10 millimeters and
had a mild reactivity rating (2) from the extract. Pseudomonas aeruginosa produced complete inhibitory activity (+++) with a total
mean zone of inhibition of 10 millimeters and had mild reactivity (2) against the pomelo peel. On the other hand, Staphylococcus
aureus produced partial inhibitory activity (++) with mean zone of inhibition of 10 millimeters and had a mild reactivity (2) against
the extract concentration.
In comparison, the antibiotic, Levofloxacin (5 ug), which served as a positive control for Escherichia coli and Pseudomo-
nas aeruginosa, produced complete inhibitory activity (+++) with a total mean zone of inhibition of 18.73 and 18.70 for Escherichia
coli and Pseudomonas aeruginosa, respectively. Both resulted to a severe reactivity rating (4).
For the positive control of the Staphylococcus aureus, a different antibiotic, Clindamycin (2 ug) was utilized. It produced
complete inhibitory activity (+++) with a total mean zone of inhibition of 15.02 and a moderate reactivity rating (3) for Staphylo-
coccus aureus.
The results of the antimicrobial activity test mean that the 75% and 95% concentration of the pomelo peel extract have the
capacity to completely inhibit the proliferation of the bacteria Escherichia coli and Pseudomonas aeruginosa. This shows that the
Journal of Health Sciences Volume 1 Issue 1 | April 2018
78 Journal of Health Sciences

pomelo peel has positive antibacterial properties. These findings grandis L. Osbeck) against fructose-mediated protein
coincide with the study of Borah (2013) which determined that oxidation and glycation. EXCLI Journal; 12, 491-502.
the plant extract of Citrus maxima showed significant antibacte-
rial activity against Escherichia coli and Pseudomonas aerugi- Cheong, M. W., Shao, Q. L., Zhou, W., and Yu, B., (2012).
nosa. Both bacteria displayed mild reactivity to the extract. Chemical composition and sensory profile of pomelo
It is to be noted, however, that the 75% concentration (Citrus grandis) juice. Food Chemistry, 135(4), 2505-
of the pomelo peel extract did not completely inhibit the pro- 13.
liferation of the Staphylococcus aureus. Hence, it may be con-
cluded that the extract is not as effective when exposed to this Keohnke, A. & Friedrich, R. (2015). Review: Antibiotic discov-
microorganism which showed mild reactivity to the pomelo peel ery in the age of structural biology - a comprehensive
extract. overview with special reference to development of
The antibiotics Levofloxacin and Clindamycin com- drugs for the treatment of Pseudomonas aeruginosa
pletely inhibited the growth of bacteria and all microorganisms infection. In Vivo, 29(2), 161-7.
tested either reacted moderately or severely to the aforemen-
tioned antibiotics. Comparing the extract with the positive con- Lan-Phi, N. & Vy, T. (2015). Chemical composition, antioxi-
trol or antibiotics, it showed that the antibiotics were still more dant, and antibacterial activities of peels’ essential oils
effective in killing and inhibiting the growth of bacteria. Upon of different pomelo varieties in the south of Vietnam.
seeing the results of the antimicrobial test, the higher concen- International Food Research Journal, 22(6), 2426-31.
tration of the extract displayed a more effective inhibition of
bacteria. Muhammad Sikander Dar, R. L., (2015). Evaluation of the
chemical constituents and the antibacterial activity
CONCLUSION AND RECOMMENDATION of essential oil of citrus karna fruit peel. Internation-
Based on the findings of the study, it was concluded that al Journal of Pharmaceutical Sciences and Research,
the pomelo peel extract showed positive results in inhibiting the 7(3), 1245-50.
growth of bacteria from the three selected microorganisms. The
two different concentrations of pomelo peel extract showcased Naradisorn, M. & Ruenkum, A. (2009). Preliminary study on an-
its capability to completely inhibit the proliferation or spread of timicrobial activity of crude extracts of pomelo albedo
Escherichia coli and Pseudomonas aeruginosa. The extract ex- against Colletotrichum gloeosporioides. Asian Journal
hibited a partial or slight inhibitory activity against Staphylococ- of Food and Agro-Industry, Special Issue, 138-142.
cus aureus. The higher concentration showed a better antibacte-
rial effect on S. aureus which implies more components of the Petkovsek, Z., Elersic, K., Gubina, M., Zgur-Bertok, D., &
pomelo peel are essential in order to increase the effectiveness Starcic-Erjavec, M. Virulence potential of Escherichia
of the antibacterial properties from the pomelo peel extract. coli isolates from skin and soft tissue infections. Jour-
nal of Clinical Microbiology, 47(6), 1811-7.
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