Green Synthesis of Silver nanoparticles by Luffa acutangula peel extract and their anti-microbial activity - Semantic Scholar
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Research Article
THE INTERNATIONAL JOURNAL OF GLOBAL SCIENCES (TIJOGS)
ISSN Print: 2663-0141; ISSN Online: 2663-015X
Vol. 2(1) Jan-March, 76-81; 2019
http://www.rndjournals.com
Green Synthesis of Silver nanoparticles by Luffa acutangula peel extract and
their anti-microbial activity
Madhavi Vemula1 , Anna Tanuja Safala1
1.Department of Chemistry, BVRITH College of Engineering for Women, Hyderabad, India.
Corresponding author: Email:madhuchem9@gmail.com Contact mobile:91-9160072989
ABSTRACT
In recent years, environmental friendly synthesis approaches and the use of benign reaction precursors are becoming more and more
popular in chemistry, chemical technologies and the need for ecological methods of synthesis is increasing to minimize or even
eliminate the use of hazardous chemicals. In this concern, “green” processes have been adopted for the benign synthesis approaches
that use less reaction conditions and non-hazardous reaction precursors. Another important advantage of green synthesis methods lies
in its cost-effectiveness and in the abundance of raw materials. Application of green chemistry to the synthesis and characterization of
nanomaterials has vital importance in medicinal and technological aspects. Synthesis of silver nanoparticles is of much interest to the
scientific community because of its unique properties and wide range of applications. The present study deals with the synthesis of
silver nanoparticles using aqueous extract of Ridge Gourd (Luffa acutangula) peel as the reducing agent to study the antimicrobial
activity. The formation of Ag-NPs was confirmed by Scanning Electron Microscopy (SEM) equipped with Energy Dispersive X-Ray
Spectroscopy (EDS), Fourier Transform-Infra Red Spectroscopy (FT-IR), X-Ray Diffraction (XRD) pattern. The FT-IR analysis
reveals that the synthesized Ag nanoparticles were capped by the polyphenolic compounds present in the peel extract. The antimicrobial
activity of these nanoparticles was studied against Staphylococcus aureus, E.coli, Pseudomonas aeruginosa, Proteus vulgaris. The
present study indicated that Ag-NPs have considerable antimicrobial activity in comparison with standard antimicrobial drug.
Keywords: Green synthesis, Ridge Gourd (Luffa acutangula), Silver nanoparticles, Antimicrobial activity
INTRODUCTION:
Due to the rapid industrialization and urbanization, our environment is threatened by a large amount of vulnerable chemicals, gases
released, and so there is a need to learn about the secrets present in the nature and its products that leads to the growth of scientific
advancements in the synthesis of nanoparticles. Nanotechnology, a multidisciplinary scientific undertaking, involves the creation and
utilization of materials, systems in the nanoscale. The field of nanotechnology is currently expected to create innovations and play a
critical role in various fronts. In recent years, scientists in the field of nanotechnology found metal nanoparticles have extensive
applications in diverse fields. The physical and chemical properties of metal nanoparticles are mainly determined by its size, shape,
composition, crystallinity and structure [1,2].
Currently, sustainability initiatives use green chemistry to reduce chemical products and processes that reduce or eliminate the use of
generation of hazardous substances and protect our global environment. As metal nanoparticles are of use in various applications i.e.,
electronics, biology and biomedical applications, material science, physics, environmental remediation fields, the development of cost
efficient and ecologically benign methods of their synthesis still remains a scientific challenge. Hence, it is vital to select stabilizing
agents and pathways that are environmental friendly to synthesize metal nanoparticles which can be non-toxic and easy to implement.
Biosynthesis of metallic nanoparticles is useful not only because of its reduced environmental impact compared with some of the
physicochemical production methods, but also because it can be used to produce large quantities of nanoparticles that are free of
contamination and have a well-defined size and morphology [3]. Nowadays the researchers are much interested in ideal synthesis of
nanoparticles which are formed at ambient temperatures, neutral pH, low costs and environmental friendly fashion. Among the
biological alternatives, plant extracts seem to be best options as they are cost effective, require little or no maintenance and they are
considered as nature’s “chemical factories”.
Nanoparticels of noble metals like gold, silver and platinum are recognized to have significant applications in electronics, magnetic,
optoelectronics and information storage. Among them silver nanoparticles being most exploited and has become the focus of much
research interest due to their unexpected applications [4-7]. Silver nanoparticles has much significance in the field of nanotechnology
because of their unique properties such as chemical stability, good conductivity, catalytic activity and most importantly antimicrobial
activities which can be incorporated into superconducting materials, electronic components, cosmetic and food industries. It has been
shown that silver nanoparticles have effective antimicrobial activity and have been applied to a wide range of health care products,
To cite this paper: Vemula, M. and A. T. Safala, 2019. Green Synthesis of Silver nanoparticles by Luffa acutangula peel extract and their anti-microbial activity. The
Intl. J. Global Sci. 2(1): 76-81
75Vemula and Safala. / The Intl. J. Global Sci., Vol. 2, No. 1 Jan-March, 2019
such as burn dressings, medical devices and water purification systems [8,9]. A lot of literature has been reported to till date on
biological syntheses of silver nanoparticles using microorganisms including bacteria, responsible for the reduction of metal compounds
to their respective nanoparticles. The microbe mediated synthesis of silver nanoparticles is not of industrial feasibility due to the
requirements of highly aseptic conditions and their maintenance. Therefore, the use of plant extracts for this purpose is potentially
advantageous due to the ease of improvement, the less biohazard as well as providing natural capping agents for the stabilization of
silver nanoparticles [10].
In this article, we describe a simple and eco-friendly method for the synthesis of silver nanoparticles by the reduction of aqueous silver
ions using Luffa peel extract and their antimicrobial activity against Staphylococcus aureus, E.coli, Pseudomonas aeruginosa, Proteus
vulgaris was studied and is compared with commercially available antimicrobial agent.
The Ridge Gourd is an extremely popular vegetable in the Asian, African and the Arabic countries. It is also known by other names
such as luffa, loofah, turiya, turai, tori etc. Ridge gourd (Luffa acutangula L. Roxb), belongs to the genus Luffa, family cucurbitaceae
it is popularly called as angled gourd, angled loofah. And has chromosome number 2n = 26. Luffein is a gelatinous substance having
several medicinal properties found in the Ridge gourd.
Luffa is an abundantly available and extremely advantageous through the health perspective. Luffa is antirheumatic and used for
meridians. It also has analgesic and hemostatic properties. Luffa is quite lower in saturated fats as well as calories. It really is abundant
with dietary fiber, vitamin C, riboflavin, zinc, thiamin, iron, as well as magnesium. It exhibits various pharmacological activities like
CNS depressant, immunomodulatory, antitumor, anti-HIV, anti-inflammatory, hepatoprotective, larvicidal and in vitro antioxidant
[11-15]. Ridge gaurd peel powder and its extracts showed slightly higher antioxygenic activity than its pulp powder and its extracts
[16]. This may be attributed to the presence of higher amounts of phenolics, flavonoids and anthocyanins which have been reported as
potential antioxidants [17]. B -Carotene is a natural product that could abundantly be found in fruits and vegetables. It is composed of
polyunsaturated hydrocarbons with two retinyl groups and it has been proven as a powerful anti-oxidant with excellent singlet oxygen
scavenging property that prevents attack of free radicals in the body [18]. Its abundance in food and non-toxic nature could be the
interest in mass production of chemically reduced Ag, which could potentially reduce the production cost as well as the toxic waste.
Thus, its deoxygenation efficacy in the synthesis of chemically reduced Ag Nps is worth to be further explored. The large amount of
active compound B -carotene present in aqueous peel extract [19] is responsible to control the shape and size of nanoparticles produced
apart from being an antimicrobial agent. The compound extracted was further used for the reduction of Ag + into Ag0 as shown in Figure
1. B –Carotene, a macrostructured antioxidant functions both as reducing and capping agent for the Ag + and stabilizes the formed Ag
Nps. This novel synthetic method method is an extremely green approach that generates bulk quantities of relatively stable Ag Nps
using aqueous peel extract of Ridge Guard at room temperature.
Figure 1 Formation of Ag nanoparticles by β-carotene, an antioxidant present in Luffa Acutangula peel extract
EXPERIMENTAL METHODS
2.1. Preparation of Luffa Acutangula peel extract
Ridge guards (Luffa Acutangula) were purchased from local market and were peeled. The peels were washed thoroughly with double-
distilled water to remove all the dust particles and dried at room temperature. The dried peels were ground and 10g of powder was
mixed with 100ml double-distilled water. This mixture was boiled and filtered through Whatman No.1 filter paper to remove particulate
matter and to get clear solution which was refrigerated (4°C) in 250ml Erlenmeyer flask for further experiment. The extract was used
as reducing agent and stabilizer. Sterile conditions were maintained at each and every step for the effectiveness and accuracy in results
without contamination.
2.2. Synthesis of AgNps
To cite this paper: Vemula, M. and A. T. Safala, 2019. Green Synthesis of Silver nanoparticles by Luffa acutangula peel extract and their anti-microbial activity. The
Intl. J. Global Sci. 2(1): 76-81
76Vemula and Safala. / The Intl. J. Global Sci., Vol. 2, No. 1 Jan-March, 2019
AgNO3 was procured from SD Fine chemicals Ltd and is used as received without further purification. AgNO 3 solution of 100ml
having 3mM strength was prepared. To this solution, 20ml of peel extract was added and stirred continuously at ambient conditions.
This resulted in colour change of the mixture from faint yellow to dark brown solution within minutes which indicates the formation
of AgNPs. Similar changes in colour have also been observed in previous studies [20-24]. The suspension of AgNPs is allowed to settle
down and the excess liquid was decanted. The resultant AgNPs were characterized using the techniques Scanning Electron Microscopy
(SEM) equipped with Energy Dispersive X-Ray Spectroscopy (EDS), Fourier Transform-Infra Red Spectroscopy (FT-IR), X-Ray
Diffraction (XRD) measurements.
1.3 Characterization Techniques
Crystalline metallic silver nanoparticles were examined using an X-ray diffractometer (Shimadzu, XRD-6000) equipped with Cu Kα
radiation source using Ni as filter at a setting of 30 kV/30 mA. All X-ray diffraction (XRD) data were collected under the experimental
conditions in the angular range 3° ≤ 2θ ≤ 50°. Fourier transform infrared (FT-IR) spectra for Ridge guard peel extract powder and silver
nanoparticles was obtained in the range 4,000 to 400 cm−1 with an IR-Prestige-21 Shimadzu FT-IR spectrophotometer, by KBr pellet
method. Scanning Electron microscopy (SEM) analysis of synthesized silver nanoparticles was done using a Hitachi S-4500 SEM
machine (Chiyoda-ku, Japan).
RESULTS AND DISCUSSION
3.1. Chemical constituents of extract
Phytochemical analysis of Luffa acutangula peel extract reveals that the extract contains polyphenols, proteins, flavonoids and
antioxidants [25]. The Ag+ reduction was based on these molecules. The large amount of active compound total carotenes present in
aqueous peel extract (14) is responsible to control the shape and size of nanoparticles produced apart from being an antimicrobial agent.
The compound extracted was further used for the reduction of Ag + into Ag0 as shown in Figure 1.
3.2. Spectral Analysis
3.2.1. FT-IR Analysis
The FT-IR spectrum obtained for the resulting Ag nanoparticles obtained by Luffa acutangula peel extract is shown in fig 2. Absorbance
bands seen at 1,563 and 1,461 cm−1 identified as amide I and amide II which arise due to carbonyl (C=O) and amine (−NH) stretching
vibrations in the amide linkages of proteins mainly involved in reduction of Ag + ions to Ag0 nanoparticles. The FT-IR spectroscopic
Figure 2 Representative FT-IR spectra of the formed Ag nanoparticles by Luffa Acutangula peel extract
study also confirmed that the protein present in Luffa acutangula peel extracts acts as a reducing agent and stabilizer for the silver
nanoparticles and prevents agglomeration. The carbonyl group of amino acid residues has a strong binding ability with metal,
suggesting the formation of a layer covering silver nanoparticles and acting as a stabilizing agent to prevent agglomeration in the
aqueous medium. The result of this FTIR spectroscopic study confirmed that Luffa acutangula peel extract has the ability to perform
dual functions of reduction and stabilization of silver nanoparticles in aqueous medium.
To cite this paper: Vemula, M. and A. T. Safala, 2019. Green Synthesis of Silver nanoparticles by Luffa acutangula peel extract and their anti-microbial activity. The
Intl. J. Global Sci. 2(1): 76-81
77Vemula and Safala. / The Intl. J. Global Sci., Vol. 2, No. 1 Jan-March, 2019
3.2.2. XRD Analysis
The exact analysis of the biosynthesized silver nanoparticles formed can be deduced from the characteristic peaks observed in the XRD
spectrum of the sample. The XRD analysis i.e., fingerprints characterization of Ag Nps crystals was conducted with Philips PW1760
and is shown in figure 3. XRD pattern of the plant derived AgNPs shows four intense peaks in the whole spectrum of 2 θ values ranging
from 20° to 60°. The XRD pattern observed may be indexed based on the face-centered crystal structure of silver. The peak at 2 θ of
28.09° marked with (220) indicates that the Ag Nps are predominantly present in the sample. The broadening of XRD peaks around
their bases indicates that silver particles are in nanosize.
Figure 3 Representive XRD pattern of stabilized Ag nanoparticles
3.2.3.SEM and EDS Analysis
SEM (model CARL-ZEISS EVO MA 15) studies were conducted to observe the shape and size of the nanoparticles synthesized using
Luffa acutangula peel extract. This instrument is used to determine the texture of crystal growth and could also be indirectly used to
measure the particle size of the materials. The representative SEM micrograph (fig 4) of synthesized Ag Nps reveals that the Ag Nps
are polydispersed with varied sizes with spherical in shape ranging from 20 to 70 nm. The analysis of Ag Nps by Energy Dispersive
Spectroscopy (EDS) confirmed the presence of the signal characteristic of elemental silver (Fig 5). All the peaks observed in the spectra
are assigned. The absorption band peak at approximately 3KeV shows the absorption of metallic Ag Nps.
Figure 4 Representative SEM image of Ag nanoparticles synthesized using Luffa Acutangula peel extract
To cite this paper: Vemula, M. and A. T. Safala, 2019. Green Synthesis of Silver nanoparticles by Luffa acutangula peel extract and their anti-microbial activity. The
Intl. J. Global Sci. 2(1): 76-81
78Vemula and Safala. / The Intl. J. Global Sci., Vol. 2, No. 1 Jan-March, 2019
Figure 5 Representative EDS image of Ag nanoparticles synthesized using Luffa Acutangula peel extract
3.3. Estimation of antibacterial activity
The antibacterial activity of the Ag Nps synthesized from the Luffa acutangula peel extract was effectively accessed against
Staphylococcus aureus, E.coli, Pseudomonas aeruginosa, Proteus vulgaris by Disc diffusion method (Table 1). Initially nutrient agar
medium was prepared, sterilized and cooled. Medium is inoculated with given organism (tubes were stirred gently to obtain even
distribution of microorganisms) and then transferred asceptically into sterile petriplates. The medium was allowed to solidify and cups
were made with the help of borer. AgNps were made into solution form and paced in caps with the help of micro pipette. Then
petriplates were kept for incubation at 37° C for 24 hrs. The inhibition zones were observed and measured for analysis against each
type of microorganism which is shown in fig 6. The results showed higher antibacterial activity against Proteus vulgaris (21.5) and
Staphylococcus aureus (18.5), whereas moderate activity was revealed against Pseudomonas aeruginosa (15) and E.coli (15).
Figure 6 Representative antimicrobial activity of Ag nanoparticles synthesized using Luffa Acutangula peel extract
Table 1: Antibacterial activity of Silver nanoparticles for selected bacteria
Name of the organism average inhibition zone
diameter(mm)
Escherichia coli 15
Proteus vulgaris 21.5
Pseudomonas aeruginosa 15
Staphylococcus aureus 18.5
CONCLUSIONS
The biological synthesis of Ag Nps using Ridge Guard (Luffa acutangula) peel extract has been reported for the first time. It was
concluded as an effective, eco-friendly and convenient green method for the synthesis of Ag Nps. Following the addition of peel extract
to the silver nitrate solution, Ag Nps began to form within 10 min at ambient conditions. Spherical, polydisperse AgNps of particle
To cite this paper: Vemula, M. and A. T. Safala, 2019. Green Synthesis of Silver nanoparticles by Luffa acutangula peel extract and their anti-microbial activity. The
Intl. J. Global Sci. 2(1): 76-81
79Vemula and Safala. / The Intl. J. Global Sci., Vol. 2, No. 1 Jan-March, 2019
sizes ranging from 20 to 70nm with the average size of 35nm are obtained. The formation of AgNps was confirmed by SEM equipped
with EDS, XRD, FT-IR spectroscopy. The antimicrobial activity of the biosynthesized Ag Nps against bacteria species Staphylococcus
aureus, E.coli, Pseudomonas aeruginosa, Proteus vulgaris was evaluated. The plant material responsible for the reduction and
stabilization of Nps needs further study including extraction and identification of the compounds presented in the extract.
ACKNOWLEDGEMENTS
The authors are thankful to the authorities of Vishnu Institute of Pharmaceutical Education and Research, Narsapur for helping us with
antimicrobial activity.
CONFLICT OF INTEREST: The authors declare that they have no conflict of interest.
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To cite this paper: Vemula, M. and A. T. Safala, 2019. Green Synthesis of Silver nanoparticles by Luffa acutangula peel extract and their anti-microbial activity. The
Intl. J. Global Sci. 2(1): 76-81
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