Global Journal of Medicinal Plant Research

2020 June; 8(1): pages 1-4

DOI: 10.22587/gjmpr.2020.8.1.1
Research Article AENSI Publications

Phytochemical Screening and Antibacterial Properties of

Silverbush (Peperomia pellucida) against Selected Cultured Bacteria
Jessica O. Tablang1, Ron Patrick C. Campos1* and James Kennard S. Jacob1
1
Department of Biological Sciences, College of Arts and Sciences, Isabela State University-Echague, Isabela, Philippines, 3309

Correspondence Author: Ron Patrick C. Campos, Department of Biological Sciences, College of Arts and Sciences, Isabela State University, San Fabian,
Echague, Isabela, Philippines,
Phone number: (+639)21-884-8521, E-mail: rpcampos023@gmail.com

Received date: 31 August 2019, Accepted date: 20 June 2020

Copyright: © 2020 Tablang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

ABSTRACT
Background: Medicinal plants are great repository of natural products that can be exploited for pharmaceutical development.
Peperomia pellucida (L.) Kunth is a widely utilized plant that is traditionally used as treatment for various illnesses. Objective: This
study aimed to determine the phytochemistry and antimicrobial activity of the crude extracts of P. pellucida. Results: Phytochemical
tests revealed that P. pellucida contains valuable phytochemical constituents namely alkaloids, flavonoids, glycosides, terpenoids and
steroids. Moreover, ethanolic extracts of P. pellucida showed antibacterial activity against E. coli and S. aureus with a notable average
inhibition of 15.43±0.67 mm and 13.22±0.34 mm respectively. Conclusion: These results affirm that P. pellucida possesses bioactive
compounds that can be potentially developed as novel drugs.

Key words: medicinal plant, Peperomia pellucida, phytochemistry, secondary metabolites, shiny bush

INTRODUCTION
The study of medicinal plants has increasingly become the focus of extensive research throughout the world mainly
because of the diversity and potential that these plants offer as source of novel natural products (Lee et al., 2008; Street, 2012).
Peperomia pellucida, also called Silverbush, belongs to the family Piperaceae. It is an herbaceous plant found in many South
American and Asian countries. The plant is distinguished by its heart-shaped and fleshy leaves with lush and succulent stems,
shallow roots and small flowers, which eventually develop into numerous tiny seeds attached on cord-like spikes (Mosango,
2008). The plant has been widely utilized in various medicinal practices and is used traditionally for treatment of different
ailments such as convulsions, conjunctivitis, headache, fever, gout, skin diseases and rheumatic pains (Mosango, 2008; Ghani,
1998).
Natural products of plant origin remain the most important sources of new drugs (Odugbemi, 2006). Studies regarding
plant phytochemistry and antimicrobial efficacy are therefore important because of their historical significance and also due to the
fact that a portion of the world still relies on plants for the treatment of different diseases (Martinez et al., 1996; Jadeja et al.,
2005). With this, the present study was carried out to determine the phytochemistry and antimicrobial activity of the crude extracts
of P. pellucida against several selected cultured microorganisms.

MATERIALS AND METHODS

Collection of plant samples

Plant samples of P. pellucida were collected from various areas within the Isabela State University-Main
Campus, Isabela, Philippines. The shoot parts of the plant were obtained, and the roots were discarded. The samples were
rinsed with distilled water and dried in open air under a shade in order to prevent the ultra violet rays from inactivating the
chemical substances present in the plant. The dried leaves were then pulverized using mechanical grinder to a fine powder and
was stored.
Preparation of Crude Extract

Global Journal of Medicinal Plant Research

ISSN:2074-0883
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Citation: Tablang et al, 2020. Phytochemical Screening and Antibacterial Properties of Silverbush (Peperomia pellucida) against Selected
Cultured Bacteria. Global Journal of Medicinal Plant Research., 8(1): 1-4. DOI: 10.22587/gjmpr.2020.8.1.1

Fifty (50) grams of powdered sample of P. pellucida was soaked on 500 mL laboratory grade 95% ethanol for
48 hours. It was then filtered using a Whatman filter paper No. 1 in an Erlenmeyer flask. The filtrate was refluxed in a rotary
evaporator until a sticky residue is obtained. The extracted product was stored and maintained in a flask.
Qualitative Phytochemical Screening
The crude extracts of P. pellucida were qualitatively screened for the presence of phytochemicals based on standard
protocols described by Khandelwal (2002) and Sofowara (1993). Results were determined based on the color and intensity of
the reaction and was interpreted as (+) if chemical is present in traceable amount and (-) if chemical is absent.
Test for alkaloids
Wagner’s test: A few ml of the extract was added with a few drops of Wagner’s reagent (1.27g of iodine and 2g of
potassium iodide dissolved in 100 ml of distilled water). The formation of reddish-brown precipitate confirms the presence of
alkaloids.

Test for saponins

Foam test: A portion of the extract was diluted with distilled water until the volume was made up to 20 ml. The solution
was vigorously shaken and the formation of persistent froth indicates presence of saponins.

Test for tannins

Braymer’s test: A fraction of the extract was added with 10% alcoholic FeCl3 solution and the formation of blue or
brown to dark green coloration indicates the presence of tannins.

Test for glycosides

Keller-Kelliani test: Five (5) ml of the extract was mixed with two (2) ml of glacial acetic acid (CH3CO2H) containing
one (1) drop of FeCl3. The mixture was carefully added to a prepared one (1) ml of concentrated H 2SO4 to form a lower layer.
The presence of a brown ring at the interface indicates presence of glycosides.

Test for terpenoids

Salkowski’s test: Two (2) ml of the extract was added with one (1) ml of chloroform followed by a few drops of
concentrated sulfuric acid. The immediate presence of reddish-brown layer indicates the presence of terpenoids.

Test for flavonoids

Alkaline Reagent test: A few drops of sodium hydroxide solution (aqueous NaOH and HCl) were added to a small
amount of the extract and observed for the formation of yellow to orange color.

Test for sterols

Liebermann–Burchard test: One (1) ml of the extract was dropped with a few ml of acetic anhydride and chloroform,
then carefully added with drops of H2SO4. The formation of dark green to red color indicates presence of sterols.

Source of test bacteria

Bacterial isolates of Escherichia coli and Staphylococcus aureus were obtained from the culture collections of the
Microbiology and Bio-Industry Laboratory, College of Arts and Sciences, Isabela State University-Echague. The bacterial isolates
were maintained in Nutrient Broth medium for the antibacterial assay.

Antibacterial Assay
The antibacterial analysis was carried out using disc diffusion method. This involved the use of filter paper discs as
carrier for the antimicrobial agents. Circular sterilized discs with 6 mm diameter was cut from Whatman no. 1 filter paper and
impregnated with liquid treatments. Cultures of E. coli and S. aureus were aseptically inoculated onto Petri plates containing
Mueller-Hinton Agar using sterile cotton swab and then the impregnated discs were placed on the surface of the medium
afterwards. The plates were incubated at 37 C and turned upside-down to prevent contamination. The zone of inhibition of each
paper disc was observed and recorded every eight (8) hours within a 24-hour incubation period. Zone of inhibition was measured
using a calibrated digital Vernier caliper. The assay was conducted in triplicates with the following treatments: distilled water
(T1), streptomycin sulfate (T2), laboratory grade 95% ethanol (T3) and ethanol extract (T4).

Statistical Analysis
Statistical analysis was laid out in a Completely Randomized Design (CRD) with three replicates per treatment. The
recorded data were treated statistically using one-way analysis of variance (ANOVA). The means were compared by Least
Significant Difference test at p< 0.05 using IBM™ SPSS v23.
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Citation: Tablang et al, 2020. Phytochemical Screening and Antibacterial Properties of Silverbush (Peperomia pellucida) against Selected
Cultured Bacteria. Global Journal of Medicinal Plant Research., 8(1): 1-4. DOI: 10.22587/gjmpr.2020.8.1.1

RESULTS AND DISCUSSION

The ethanolic extract of P. pellucida was used to determine the phytochemical constituents of the plant. The ethanolic
extracts of the plant indicated presence of the different phytochemical constituents (Table 1). The results indicate that P. pellucida
contains valuable phytochemical constituents namely alkaloids, flavonoids, glycosides, terpenoids and steroids. However, the
results also suggest the absence of saponins and tannins.

Table 1. Phytochemical constituents of P. pellucida crude extract.

PHYTOCHEMICAL SCREENING

Constituents Result

Alkaloids +

Saponins -

Flavonoids +

Glycosides +

Terpenoids +

Tannins -

Steroids +

Legend: (+) means presence of the chemical; (-) means absence of the chemical

A handful of studies have elucidated the several phytochemical constituents of the plant. One study has isolated four
chemicals from P. pellucida namely alkaloids, cardenolides, saponins and tannins by using dried powdered samples of the plant
(Egwuche et al., 2011). On the other hand, saponins were believed to be selectively present only in the leaf extracts of the plant
(Majumder, 2012). Discrepancy of results obtained in the phytochemical screening can be attributed to the difference in the
extraction method. The results show that P. pellucida contains pharmacologically active compounds which can be utilized in a
wide array of pharmaceutical applications.
The presence of bioactive compounds in P. pellucida provides the plant with various effects on biological activities. The
zones of inhibition of the different treatments against E. coli and S. aureus is presented in Table 2.

Table 2. Antibacterial activities of P. pellucida crude extract against E. coli and S. aureus.
Zone of Inhibition (mm)
Treatment
E. coli S. aureus
a
Distilled water 6.00±0.00 6.00±0.00a

Streptomycin 33.07±2.98b 32.20±1.72b

Ethanol 6.97±0.42a 8.13±0.47a

P. pellucida extract 15.43±0.67c 13.22±0.34c

Note: Means in the same column not sharing the same superscript are significantly different at 5% significance level.

The results indicate the presence of bioactive compounds in the P. pellucida extracts that can inhibit the growth of both
E. coli and S. aureus. The antibacterial drug streptomycin, treated as positive control, inhibited the E. coli with a diameter of
33.07±2.99 mm and S. aureus with 32.20±1.72 mm. The ethanolic extracts of P. pellucida recorded an average diameter of
15.43±0.67 for the E. coli and 13.22±0.34 for the S. aureus. Ethanol exuded small zones of inhibition with 6.97±0.42 and
8.13±0.47 for the E. coli and S. aureus respectively. The negative control distilled water produced no zone of inhibition. Statistical
analysis revealed that the ethanolic extracts of P. pellucida were significantly different with the zones of inhibition produced by
distilled water, ethanol and streptomycin.
The antibacterial properties of P. pellucida has been investigated by several authors and results indicated the presence of
inhibiting properties in the plant. The plant has been determined to possess broad spectrum of antimicrobial activity against
different strains of pathogenic bacteria and fungi namely E. coli, S. aureus, Bacillus subtilis, Pseudomonas aeruginosa,
4
Citation: Tablang et al, 2020. Phytochemical Screening and Antibacterial Properties of Silverbush (Peperomia pellucida) against Selected
Cultured Bacteria. Global Journal of Medicinal Plant Research., 8(1): 1-4. DOI: 10.22587/gjmpr.2020.8.1.1

Klebsiellae pneumonae, Salmonellae typhi, Candida albicans, Penicillum notatum, Rhizopus stolonifer and Aspergillus niger
(Oleyede et al., 2011). These results are further corroborated by multiple studies involving the antibacterial potential of the plant
(Wei et al., 2011). It has also been claimed that the chemical constituent dillapiol may be one of the agents that are responsible for
its antimicrobial activity (Rafael et al., 2008; Brazao et al., 2014). Phytol was believed to be a major compound present in the
tissues of P. pellucida and is one of the most important diterpenes that possesses antimicrobial properties (Kumar et al., 2010).
The results indicate the presence of antibacterial agents in the plant despite the variance in the method of extraction of the
chemical constituents. It is clear that the ethanolic extract of P. pellucida possessed antibacterial properties which can be tapped
for exploitation in different pharmacological applications.

CONCLUSIONS

Based on the foregoing results, the ethanolic extract of P. pellucida possessed phytochemical constituents that can be
utilized for the isolation of important bioactive compounds. The isolation of bioactive compounds can provide a novel and
economic way of obtaining important chemicals that possess various pharmaceutical applications. The presence of antibacterial
properties within the plant can also be of importance for the development of new therapeutic agents.
ACKNOWLEDGEMENT

The authors are grateful for the provision of materials and facilities by the Department of Biological Sciences of Isabela
State University.

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