|Year : 2021 | Volume
| Issue : 1 | Page : 35-38
Green synthesis of silver nanoparticles using Phyllanthus amarus Seeds and their antibacterial activity assessment
Jerrine Joseph1, Keren S Deborah1, R Raghavi1, A Mary Shamya1, Wilson Aruni2
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 Submission||21-Jul-2020|
|Date of Acceptance||25-Sep-2020|
|Date of Web Publication||13-Mar-2021|
Dr. Jerrine Joseph
Scientist -D, Centre for Drug Discovery and Development, Sathyabama Institute of Science and Technology, Chennai -600 119
Source of Support: None, Conflict of Interest: None
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 27];5:35-8. Available from: https://www.bmbtrj.org/text.asp?2021/5/1/35/311086
| Introduction|| |
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., 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. 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.
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. 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.
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.,,,, 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.
| Methods|| |
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.
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.
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.
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.
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.
| Results|| |
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].
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 in [Figure 3].
|Figure 3: UV-VIS spectroscopy for silver nanoparticles synthesized using Phyllanthus amarus seed extract|
Click here to view
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|
Click here to view
The extracts of P. amarus seeds showed potent antimicrobial activity against Gram-positive and Gram-negative bacteria represented in [Table 2].
|Table 2: Antibacterial activity of silver nanoparticles using Phyllanthus amarus seed extract against pathogens|
Click here to view
| Discussion|| |
The green synthesis of AgNPs has received significant attention in the growing environment (Oza et al. 2012). 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). 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). 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. 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. 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|| |
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
Conflicts of interest
There are no conflicts of interest.
| References|| |
Dos Santos CA, Seckler MM, Ingle AP, Gupta I, Galdiero S, Galdiero M, et al
. Silver nanoparticles: therapeutical uses, toxicity, and safety issues. Journal of pharmaceutical sciences 2014;103:1931-44. https://doi.org/10.1002/jps.24001
Pugazhendhi A, Edison TNJI, Karuppusamy I, Kathirvel B. Inorganic nanoparticles: A potential cancer therapy for human welfare. International Journal of Pharmaceutics 2018;539:104–11. https://doi.org/10.1016/j.ijpharm.2018.01.034
Shabir Ahmad, Sidra Munir, Nadia Zeb, Asad Ullah, Behramand Khan, Javed Ali, Green nanotechnology: a review on green synthesis of silver nanoparticles an ecofriendly approach, Int J Nanomedicine 2019;14: 5087–5107. doi: 10.2147/IJN.S200254
Khan ST, Musarrat J, Al-Khedhairy AA. Countering drug resistance, infectious diseases, and sepsis using metal and metal oxides nanoparticles: Current status. Colloids and surfaces B, Biointerfaces 2016;146:70–83. https://doi.org/10.1016/j.colsurfb.2016.05.046
Patel J. R, Tripathi P, Sharma V, Chauhan N. S, Dixit V. K, Phyllanthus amarus
: ethnomedicinal uses, phytochemistry and pharmacology: a review. J. Ethnopharmacol 2011;138:286–313. https://doi.org/10.1016/j.jep.2011.09.040
He Y, Wei F, Ma Z, Zhang H, Yang Q, Yao B, et al
. Green synthesis of silver nanoparticles using seed extract of Alpinia katsumadai, and their antioxidant, cytotoxicity, and antibacterial activities. RSC advances 2017;7:39842-51. DOI: 10.1039/C7RA05286C.
Singh K, Panghal M, Kadyan S, Chaudhary U, Yadav JP. Green silver nanoparticles of Phyllanthus amarus: as an antibacterial agent against multi drug resistant clinical isolates of Pseudomonas aeruginosa. Journal of nanobiotechnology. 2014;12:1-9. https://doi.org/10.1186/s12951-014-0040-x
Joshi H, Parle M. Pharmacological evidences for antiamnesic potentials of Phyllanthus amarus in mice. African Journal of Biomedical Research 2007;10:1119–5096.
Alagan A, Jantan I, Kumolosasi E, Ogawa S, Abdullah MA, Azmi N. Protective effects of Phyllanthus amarus against lipopolysaccharide-induced neuroinflammation and cognitive impairment in rats. Frontiers in pharmacology 2019;10:632. doi.org/10.3389/fphar.2019.00632.
N. Thirumagal, A. Pricilla Jeyakumari, Green Synthesis and Antibacterial Activity of Silver Nanoparticles (AgNPs) using Psoralea corylifolia Seed Extract. International Journal of Recent Technology and Engineering (IJRTE) 2020 8 (5).ISSN: 2277-3878. DOI:10.35940/ijrte.D4251.018520.
Ansari MA, Alzohairy MA. One-pot facile green synthesis of silver nanoparticles using seed extract of Phoenix dactylifera and their bactericidal potential against MRSA. Evidence-Based Complementary and Alternative Medicine 2018, vol. 2018, 9 Article ID 1860280. https://doi.org/10.1155/2018/1860280
Ali ZA, Yahya R, Sekaran SD, Puteh R. Green synthesis of silver nanoparticles using apple extract and its antibacterial properties. Advances in Materials Science and Engineering. 2016. https://doi.org/10.1155/2016/4102196
Rautela A, Rani J, Das MD. Green synthesis of silver nanoparticles from Tectona grandis seeds extract: characterization and mechanism of antimicrobial action on different microorganisms. Journal of Analytical Science and Technology 2019;10:1-0. https://doi.org/10.1186/s40543-018-0163-z
Krithiga N, Rajalakshmi A, Jayachitra A. Green synthesis of silver nanoparticles using leaf extracts of Clitoria ternatea and Solanum nigrum and study of its antibacterial effect against common nosocomial pathogens. Journal of Nanoscience.2015. https://doi.org/10.1155/2015/928204
Okafor F, Janen A, Kukhtareva T, Edwards V, Curley M. Green synthesis of silver nanoparticles, their characterization, application and antibacterial activity. International journal of environmental research and public health 2013;10:5221-38. https://doi.org/10.3390/ijerph10105221
Khan MZ, Tareq FK, Hossen MA, Roki MN. Green synthesis and characterization of silver nanoparticles using Coriandrum sativum leaf extract. Journal of Engineering Science and Technology 2018;13:158-66.
Saba Pirtarighat, Maryam Ghannadnia & Saeid Baghshahi, Green synthesis of silver nanoparticles using the plant extract of Salvia spinosa
grown in vitro
and their antibacterial activity assessment, Journal of Nanostructure in Chemistry 2019;9:1–9. https://doi.org/10.1007/s40097-018-0291-4
Oza G, Pandey S, Shah R, Sharon M. Extracellular fabrication of silver nanoparticles using Pseudomonas aeruginosa and its antimicrobial assay. Adv Appl Sci Res 2012;3:1776-83. ISSN: 0976-8610
Ahmed S, Saifullah, Ahmad M, Swami BL, Ikram S. Green synthesis of silver nanoparticles using Azadirachta indica aqueous leaf extract. Journal of radiation research and applied sciences. 2016;9:1-7. https://doi.org/10.1016/j.jrras.2015.06.006
Abolaji AO, Adebayo HA, Odesanmi OS. Nutritional qualities of three medicinal plant parts (Xylopia aethiopica, Blighia sapida and Parinari polyandra) commonly used by pregnant women in the western part of Nigeria. Pakistan Journal of Nutrition 2007;6:665-8. ISSN 1680-5194.
Ilangkovan M, Jantan I, Mohamad HF, Husain K, Razak A, Faiz A. Inhibitory effects of standardized extracts of Phyllanthus amarus and Phyllanthus urinaria and their marker compounds on phagocytic activity of human neutrophils. Evidence-Based Complementary and Alternative Medicine. 2013. https://doi.org/10.1155/2013/603634
Lara HH, Ayala-Núñez NV, Turrent LD, Padilla CR. Bactericidal effect of silver nanoparticles against multidrug-resistant bacteria. World Journal of Microbiology and Biotechnology 2010;26:615-21. https://doi.org/10.1007/s11274-009-0211-3
[Figure 1], [Figure 2], [Figure 3], [Figure 4]
[Table 1], [Table 2]