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 Table of Contents  
ORIGINAL ARTICLE
Year : 2021  |  Volume : 5  |  Issue : 1  |  Page : 35-38

Green synthesis of silver nanoparticles using Phyllanthus amarus Seeds and their antibacterial activity assessment


1 Centre for Drug Discovery and Development, Sathyabama Institute for Science and Technology (Deemed to be University), CA, USA
2 California University of Science and Medicine, School of Medicine; Musculoskeletal Disease Research Laboratory US Department of Veteran Affairs; Division of Microbiology and Molecular Genetics, School of Medicine, Loma Linda University, Loma Linda, California 92350, USA

Date of Submission21-Jul-2020
Date of Acceptance25-Sep-2020
Date of Web Publication13-Mar-2021

Correspondence Address:
Dr. Jerrine Joseph
Scientist -D, Centre for Drug Discovery and Development, Sathyabama Institute of Science and Technology, Chennai -600 119
USA
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/bbrj.bbrj_139_20

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  Abstract 


Background: Green synthesis of nanoparticles has been gaining popularity due to its advantages over chemical synthesis. In the present study, silver nanoparticles (AgNPs) were synthesized by using an aqueous solution of Phyllanthus amarus leaves extract as a reducing agent. The synthesized nanoparticles were characterized using the spectroscopic techniques. The Fourier-transform infrared (FTIR) study confirmed that the seed extract also stabilized the surface of the AgNPs by acting as a capping agent. Moreover, the antibacterial activity of the plant NPs was also assessed. The synthesized nanoparticles as well as P. amarus plant extract were separately tested to examine their antibacterial activities. The activities were tested against various microorganisms, including Escherichia coli, Bacillus subtilis, Klebsiella pneumoniae, and Staphylococcus aureus. The main aim of the present study is to evaluate the green synthesis of nanoparticles using P. amarus seeds and their antibacterial activity assessment. Method: Collection and preparation of seed extract, synthesis of AgNPs, characterization of AgNPs using ultraviolet-visible (UV-Vis) absorbance spectroscopy and Fourier transforms infrared spectroscopy, determination of antibacterial activity using pathogens. All in vitro assay data signify the mean ± standard deviation of triplicates was calculated by using the MS word document. Results: The reduction of silver nitrate using the plant leaf extract was viewed by the color change in the reaction solutions. The maximum absorbance peak was seen at 400 nm for P. amarus seed extract using UV-Vis spectroscopy and FTIR measurements were carried out for the AgNPs synthesized by the plant extracts. The extracts of P. amarus seeds showed potent antimicrobial activity against Gram-positive and negative bacteria. Conclusions: The biosynthesized AgNPs using P. amarus seed extract proved to be excellent agent against pathogens. The present study showed a simple, rapid, and economical route to synthesize AgNPs. The use of P. amarus has the added advantage that this seed can be used by nanotechnology processing industries.

Keywords: Antibacterial activity, green synthesis, Phyllanthus amarus, silver nanoparticle


How to cite this article:
Joseph J, Deborah K, Raghavi R, Mary Shamya A, Aruni W. Green synthesis of silver nanoparticles using Phyllanthus amarus Seeds and their antibacterial activity assessment. Biomed Biotechnol Res J 2021;5:35-8

How to cite this URL:
Joseph J, Deborah K, Raghavi R, Mary Shamya A, Aruni W. Green synthesis of silver nanoparticles using Phyllanthus amarus Seeds and their antibacterial activity assessment. Biomed Biotechnol Res J [serial online] 2021 [cited 2021 Nov 30];5:35-8. Available from: https://www.bmbtrj.org/text.asp?2021/5/1/35/311086




  Introduction Top


Nanotechnology is the synthesis of particles with at least one dimension in the range of 1–100 nm, resulting in high surface to volume ratios. As the particle size decreases, not only does the ratio of surface area to volume increase but also the physical, chemical, and biological properties of the particles differ compared to their bulk counterparts.[1],[2] In recent years, the interest in the synthesis and properties of noble metal nanoparticles such as gold, silver, and platinum has been attracting attention in nanomedicine.[3] Silver nanoparticles (AgNPs) are widely used because of their unique properties and promising applications including pharmaceutics, agriculture, water detoxification, air filtration, textile industries, and as a catalyst in oxidization reactions.[4]

Different synthesis methods developed for nanoparticle synthesis are physical, chemical, and green synthesis. Physical methods require costly equipment, high temperature, and high pressure. In the synthesis of nanoparticles with chemical methods, toxic chemicals are used which can cause serious damage to the environment and to the livings. Due to these disadvantages, the use of physical and chemical methods is limited.[5] These methods are replaced by green synthesis which is a more environmentally friendly and cheaper method. Plants, bacteria, fungi, algae, etc., are widely used for the green synthesis of nanoparticles.[6]

Phyllanthus amarus belongs to the Euphorbiaceae family and is traditionally used for kidney ailments, diabetes, pain, jaundice, gonorrhea, chronic dysentery, skin ulcer, and hepatitis B. Recently, the plant has received increasing attention and has been studied for various pharmacological properties such as immunomodulatory, antinociceptive, anti-inflammatory, antioxidant, antibacterial, anticancer, antiulcer, gastroprotective, antifungal, antiplasmodic, antiviral, aphrodisiac, contraceptive, hepatoprotective, antihyperglycemic, antilipidemic, nephroprotective, and anti-amnesic activities.[7],[8],[9],[10],[11] The classification of Phyllanthus amarus is given in [Table 1]. The main purposes of this work are evaluating the potential of the seed extract of P. amarus for the biosynthesis of AgNPs and investigation of their antibacterial activities against both Gram-positive and Gram-negative species of bacteria.
Table 1: Classification of Phyllanthus amarus

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  Methods Top


Collection and preparation of seed extract

The seeds of Phyllanthus amarus (Amla) were collected from Koyambedu market, Chennai. The surface of date seeds was washed twice with distilled water. Ten g of seeds was shade dried and powdered using mixer and was boiled with 100 ml distilled water at 80°C for 20 min. The extract was filtered by Whatmann Filter paper. The filtrate extract was stored at 4°C and used as reducing and stabilizer agent for the synthesis of AgNPs.[12]

Green synthesis of silver nanoparticles

In a typical reaction procedure, 10 ml of seed extract was added to 90 ml of 10−3 (M) aqueous silver nitrate solution. The flask (aqueous) was then incubated at the room temperature for overnight. Any color changes of the solution were observed.[13]

Characterization of silver nanoparticles

Ultraviolet-visible absorbance spectroscopy

The formation and stability AgNPs were carried out by measuring the ultraviolet-visible (UV-vis) spectra of the solutions after diluting the sample. Distilled water was used as a blank solution. The absorbance spectra of AgNPs solution were recorded at the wavelength ranging from 200 to 800 nm by UV-Vis spectrophotometer.[14]

Fourier transforms infrared spectroscopy

The functional groups on AgNPs were validated with Fourier-transform infrared (FTIR) spectroscopy using in the range of 400–4000 cm−1.[15]

Determination of antibacterial activity

The antibacterial assays were assessed on Gram-positive and Gram-negative pathogens such as Bacillus subtilis, Escherichia coli, Klebsiella aerogenes, and Staphylococcus aureus by using the standard well-diffusion method. The antibacterial activity was measured based on the inhibition zone around the well impregnated with plant extract and synthesized silver nanoparticle.[9]


  Results Top


The reduction of silver nitrate using the plant leaf extract was viewed by the color change in the reaction solutions represented in [Figure 1] and [Figure 2].
Figure 1: Amla Seed Powder

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Figure 2: Seed Extract + Ag NO3 solution

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Visual observation and ultraviolet-visible spectroscopy

The maximum absorbance peak was seen at 400 nm for Phyllanthus amarus seed extract. It is generally recognized that UV-Vis spectroscopy could be used to examine the size and shape-controlled nanoparticles in aqueous suspensions[16] in [Figure 3].
Figure 3: UV-VIS spectroscopy for silver nanoparticles synthesized using Phyllanthus amarus seed extract

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Fourier-transform infrared analysis

FTIR measurements were carried out to identify the possible biomolecules responsible for the capping and efficient stabilization of the AgNPs synthesized by the plant extracts.[17 & 18] Absorbance bands of P. amarus were observed at 3344.42 cm−1 assigned to O–H (s) stretch, 1636.92 cm−1 assigned to C = C aromatic stretch, 1089.68 cm−1 assigned to C–N amines stretch, and 695.95 cm−1 assigned to C–H alkenes stretch, as shown in [Figure 4].
Figure 4: FTIR spectrum of silver nanoparticles synthesized by using the seed extract of Phyllanthus amarus

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Antibacterial activity

The extracts of P. amarus seeds showed potent antimicrobial activity against Gram-positive and Gram-negative bacteria represented in [Table 2].[19]
Table 2: Antibacterial activity of silver nanoparticles using Phyllanthus amarus seed extract against pathogens

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  Discussion Top


The green synthesis of AgNPs has received significant attention in the growing environment (Oza et al. 2012).[20] The phytochemicals are responsible for the antibacterial properties. The seed as well as the plant contains high amount of carbohydrate and low amount of fat and protein (Hasan et al. 2011, Abolaji et al. 2007). [21, 22 & 23] The plant-derived products do not have any toxic content and would become an effective antibacterial agent for controlling microorganisms (Ahmad et al. 2001).[24] The main mechanism behind this process is plant-assisted reduction due to the presence of the phytochemicals. The extract of P. amarus contains mainly phyllanthin, hypophyllanthin, phyltertralin, and other phytochemicals (Yuandani et al. 2013).[25] In our study, the seed extract of P. amarus showed a strong peak at 420 nm the same report was reported by Singh et al.[9] 2014.FTIR confirms the presence of different functional groups absorb characteristic frequencies of IR radiations. These biosynthesized nanoparticle has the ability to act against microorganism is mainly due to the bacterial cells contact with silver absorb silver ions, which inhibit several functions in the cell and damage the cells. Many studies state that AgNPs of P. amarus were found to be good antibacterial agent. In the current study, these nanoparticles showed good activity toward Gram-positive and Gram-negative bacteria. Lara et al.[26] reported the antibacterial activity of AgNPs against multidrug-resistant P. aeruginosa, E. coli, Streptococcus sp., and S. pyogens. Finally, the current study clearly indicates that the P. amarus extract-mediated AgNPs exhibited the excellent antimicrobial activity against bacterial pathogens.


  Conclusions Top


A critical need in the field of nanotechnology is the development of a reliable and eco-friendly process for the synthesis of metallic nanoparticles. Seeds of P. amarus are easily available. AgNPs play a profound role in the field of biology and medicine due to their attractive physiochemical properties. In the present study, we have demonstrated that the use of a natural, low-cost biological reducing agent and P. amarus seed extracts can produce metal nanostructures, through efficient green nanochemistry methodology, avoiding the presence of toxic solvents and waste. The biosynthesized AgNPs using P. amarus seed extract proved to be excellent against pathogens. The antimicrobial activity is well demonstrated by the well-diffusion method. The present study showed a simple, rapid, and economical route to synthesize AgNPs. The use of P. amarus has the added advantage that this seed can be used by the nanotechnology processing industries.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1], [Table 2]



 

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