Biocatalysis and Agricultural Biotechnology 15 (2018) 311–316

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Biocatalysis and Agricultural Biotechnology

journal homepage: www.elsevier.com/locate/bab

Synthesis of silver nanoparticles (AgNPs) from cinnamon against bacterial T

pathogens
⁎
J. Premkumara, , T. Sudhakara, Abhishek Dhakala, Jeshan Babu Shresthaa, S. Krishnakumara,
P. Balashanmugamb
a
Department of Biomedical Engineering, School of Bio & Chemical Engineering, Sathyabama Institute of Science & Technology, Chennai 600119, India
b
CHORD Division, CSIR, CLRI, Chennai, India

A R T I C LE I N FO A B S T R A C T

Keywords: Nanotechnology can be useful in diagnostic techniques, drug delivery, sunscreens, antimicrobial sanitizer and a
Cinnamon friendly manufacturing process that reduce waste products. Development of Green Nanotechnology is generating
Silver nanoparticles interest of researchers towards eco-friendly biosynthesis of nanoparticles. In the research study silver nano-
Characterization particles were synthesized from aqueous silver nitrate (1 mM) through a simple and eco-friendly method using
Antimicrobial activity
cinnamon as reductant and stabilizer. The aqueous silver ions when exposed to leaf broth were reduced and
resulted in the green synthesis of silver nanoparticles ranges from 20 nm to 30 nm. The bio reduced silver
nanoparticles were characterized by UV–Vis spectrophotometer, X ray diffraction, FESEM with EDAX and
Fourier transform infra red (FTIR) spectroscopy. The FTIR study was carried out to recognize the possible
biomolecules responsible for proficient stabilization of silver nanoparticles and their antimicrobial activity was
screened against both gram positive, gram-negative microorganisms. The method can be used for rapid and eco-
friendly biosynthesis of stable silver nanoparticles of size range from 20 to 30 nm possessing antimicrobial
activity suggesting their possible application in the field of medicine.

1. Introduction nanoparticles – when added to water paints or cotton fabrics – show a

pronounced antibacterial/antifungal effect. It was shown that smaller
Nanotechnology is a field that is mushrooming, making an impact in silver nanoparticles have a greater antibacterial/antifungal efficacy.
all spheres of human life. Nano biotechnology represents an economic The paper also provides a review of scientific literature with regard to
alternative for chemical and physical methods of nanoparticles (NP) recent developments in the field of toxicity of silver nanoparticles and
formation. The current literature survey reveals that the NP synthesis its effect on environment and human health (Anuradha et al., 2010).
using marine plants, microorganisms and algae as source has been This is a first and novel report on extracellular silver nanoparticles
unexplored and under exploited. The development of green processes synthesis using young leaf aqueous extract of Sterculia foetida as a re-
for the synthesis of NP is evolving into an important branch of nano- ducing agent. The synthesized AgNPs characterized by using spectral
technology. It has many advantages such as, ease with which the pro- analysis like UV–vis spectroscopy, Transmission Electron Microscopy,
cess can be scaled up, economic viability, etc. Presently, the researchers X-ray diffraction studies, energy dispersive X-ray, Fourier transform
are looking into the development of cost-effective procedures for pro- infrared spectroscopy. Antimicrobial activity of AgNPs was performed
ducing reproducible, stable and biocompatible AgNPs. Antibiotic re- by agar well diffusion technique. The appearance, size, and shape of the
sistance is the world's major public healthcare problem. AgNPs particles silver nanoparticles were understood by UV–vis spectroscopy, trans-
play a vital role in nano biotechnology as biomedicine against Drug- mission electron microscopy. The X-ray diffraction studies, energy
resistant bacteria (Majdalawieh et al., 2014). dispersive X-ray analysis indicate that particles are crystalline in nature.
The bactericidal effect of silver nanoparticles obtained by a novel Fourier transform infrared spectroscopy analysis revealed that the na-
electrochemical method on Escherichia coli, Staphylococcus aureus, noparticles are covered with bio-moieties on their surface. In the re-
Aspergillus niger and Penicillium phoeniceum cultures has been studied. search work, the bio functionalized silver nanoparticles thus produced
The tests conducted have demonstrated that synthesized silver have shown admirable antimicrobial effect. The synthetic procedure

⁎
Corresponding author.
E-mail address: [email protected] (J. Premkumar).

https://doi.org/10.1016/j.bcab.2018.06.005
Received 9 February 2018; Received in revised form 26 April 2018; Accepted 11 June 2018
Available online 12 June 2018
1878-8181/ © 2018 Elsevier Ltd. All rights reserved.
J. Premkumar et al. Biocatalysis and Agricultural Biotechnology 15 (2018) 311–316

involved is eco-friendly, simple and hence high range production of the

same can be considered for using them in many pharmaceutical ap-
plications (Brandt et al., 2012).
Silver is a metal known for its broad spectrum antimicrobial activity
against Gram positive and Gram negative bacteria, fungi, protozoa and
certain viruses. The persistence of antibiotic resistant bacteria has
exploited the antimicrobial properties of silver and silver-based com-
pounds, including AgNPs (Jena et al., 2012). The mechanism of cellular
toxicity exhibited by metal nanoparticles is through the release of Re-
active Oxygen Species (ROS) (Mallikarjuna et al., 2011). The anti- CINNAMON Powdered Cinnamon
bacterial properties of AgNPs are associated with its slow oxidation and
Fig. 1. Dried cinnamon bark used for the synthesis of silver nanoparticle.
liberation of Ag+ ions to the environment making it an ideal biocidal
agent. Moreover, the small size of these particles facilitates the pene-
tration of these particles through cell membranes to affect intracellular
processes from inside.
A significant feature of nanomaterials is the presence of a high
surface to volume ratio (S/V), as noted previously. Study of pure ma-
terials for significant times under the high S/V conditions becomes
problematical, even under relatively high vacuum conditions. The en-
trance of even small amounts of air into the vacuum chamber results in
almost instant oxidation of the material at the surface. To circumvent
this oxidation problem, researchers have used the noble metals ex-
tensively, starting with examination of the nucleation and growth Fig. 2. Synthesis of silver nanoparticles (control & test).
processes in the synthesis of colloidal gold (Geethalakshmi and Sarada,
2010).
The recent development and implementation of new technologies
have led to new era, the nano-revolution which unfolds role of plants in
bio and green synthesis of nanoparticles which seem to have drawn
quite an unequivocal attention with a view of synthesizing stable na-
noparticles. Although nanoparticles can be synthesized through array of
conventional methods biological route of synthesizing are good com-
petent over the physical and chemical techniques. Green principle route
of synthesizing have emerged as alternative to overcome the limitation
of conventional methods among which plant and microorganisms are
majorly exploited. Employing plants towards synthesis of nanoparticles (Before stability=412 nm) (After stability=423.2 nm)
are emerging as advantageous compared to microbes with the presence
of broad variability of bio-molecules in plants can act as capping and Fig. 3. UV spectroscopy analysis of silver nanoparticles.
reducing agents and thus increases the rate of reduction and stabiliza-
tion of nanoparticles. Biological synthesized nanoparticles have up-
surge applications in various sectors. Hence the present study envisions
on biosynthesis of nanoparticles from plants which are emerging as
nanofactories (Subashini and Kannabiran, 2013).
In recent years, plant-mediated biological synthesis of nanoparticles
is gaining importance due to its simplicity and eco-friendliness
(Geethalakshmi and Sarada, 2010). Although biosynthesis of gold na-
noparticles by plants such as alfalfa, Aloe Vera (Singhal et al., 2011),
have been reported, the potential of the plants as biological materials
for the synthesis of nanoparticles is yet to be fully explored.
Cinnamon is a powerful spice that has been used medicinally
around the world for thousands of years. According to researchers, out
of twenty-six of the most popular herbs and medicinal spices in the
world, cinnamon in terms of its protective antioxidant levels.
The metallic nanoparticles have great attention of Chemists; Physics
Development of green nanotechnology is generating interest of re-
searchers toward eco-friendly biosynthesis of nanoparticles. These
biosynthesized nanoparticles were characterized with the help of Fig. 4. FTIR analysis of synthesized silver nanoparticles.
UV–vis spectrophotometer, Atomic Absorption Spectroscopy (AAS),
Dynamic light scattering (DLS), X-ray diffraction (XRD), Fourier Table 1
transform infrared spectroscopy (FTIR), and Transmission electron Absorption peaks and corresponding vibrations of FTIR analysis.
microscopy (TEM). Stability of bio reduced silver nanoparticles was S.no. IR spectrum Types of vibration
analysed using UV–vis absorption spectra, and their antimicrobial ac-
tivity was screened against both gram-negative and gram-positive mi- 1 3259.29 O-H stretch carboxylic acid
croorganisms. Thus, this method can be used for rapid and eco-friendly 2 1634.60 C=N stretch oxime
3 564.12 C-Cl stretch Halo compound
biosynthesis of stable silver nanoparticles of size range 4–30 nm pos-
4 520.32 C-Br stretch Halo compound
sessing antimicrobial activity suggesting their possible application in 5 510.10 C-I stretch Halo Compound
medical industry (Singhal et al., 2011). In this present study silver

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Fig. 5. FESEM analysis & silver nanoparticles.

Fig. 6. XRD analysis synthesized nanoparticles.

Staphylococcus aureus Bacillus cereus Escherichia coli Pseudomonas aeruginosa

Fig. 7. Different extract samples.

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Pseudomonas aeruginosa Bacillus cereus Escherichia coliBacillus Staphylococcus aureus

Fig. 8. Zone of inhibition in different concentrations of silver nanoparticles.

collected from local grocery shop in raw form. From the components
cinnamon was selected and air dried in hot air oven then grinded to fine
powder in 25 ml of distilled water and prepared 5 different solutions
kept for stirring for 25 min and heat the solution in water bath for
30 min maintaining temperature of 50–60 °C (Figs. 1 and 2). After
cooling the solutions, it is filtered using a Whatmann filter paper and
the filtrate is stored in storage tube. The extracts are kept in refrigerator
at room temperature.

2.2. Synthesis of silver nanoparticles

Graph 1. Reaction of bacterial strands against source extracts different con- The synthesis of silver nanoparticles from all the extracts of the 5
centrations of silver nanoparticles from cinnamon. sources is subjected to the test. Then the best silver nanoparticles is
taken into the account for further tests. The productions of silver na-
noparticles are divided into 3 phases based on the ratio of the extract to
the distilled water quantity. Phase 1(1:9 ratio)-1 ml of the extracts are
taken separately in 10 different test tubes maintain both control and
test tubes for each extract. Then in each test tube 9 ml of pure distilled
water was poured, thus maintaining the ratio of 1:9. Then in each test
sample 20 µl of Silver Nitrate (AgNO3) Solution is introduced and is left
to incubate at 37 °C for 48 h for nanoparticles production. Phase 2 (1:4
ratio) Then in each test tubes 4 ml of pure distilled water was poured,
thus maintaining the ratio of 1:4. Phase 3 (0.5:4.5 ratio) Then in each
test tubes 4 ml of pure distilled water was poured, thus maintaining the
ratio of 0.5:4.5. Analysis from each phase: After 48 h of incubation in
dark environment each samples are analysed. Then after visual analysis
the ratio of 0.5:4.5 are taken into consideration. Out of all extract
cinnamon has given best production of silver nanoparticles and sub-
Graph 2. Reaction of bacterial strands with different concentrations of silver jected to characterization (Sudhakar et al., 2014).
nanoparticles.

nanoparticles were synthesized from aqueous silver nitrate (1 mM) 2.3. Characterization
through a simple and eco-friendly route using leaf broth of Cinnamon as
reductant and stabilizer. A characterization can be defined as the analysis of the inner
structure of any materials by enhancing or including the external
2. Materials and methods structure of same material. The 5 different types of characterization are
as follows – UV–VIS Spectroscopy analysis, Fourier Transform Infrared
2.1. Sample collection and extract preparation radiation (FTIR), Field emission Scanning Electron Microscope
(FESEM), X-Ray Diffraction (XRD).
Turmeric, Ginger, Cardamom, Black pepper, Cinnamon were

Table 2
Table showing zone of inhibition of synthesized AgNps from cinnamon.
Pathogens AgNps 10 µl AgNps 20 µl AgNps 30 µl AgNps 40 µl AgNps 50 µl AgNps 60 µl Positive control

Staphylococcus aureus 11 ± 0.3 12 ± 0.3 13 ± 0.3 16 ± 0.3 18 ± 0.3 19 ± 0.3 20 ± 0.3

Escherichia coli 6 ± 0.3 7 ± 0.3 9 ± 0.3 10 ± 0.3 12 ± 0.3 13 ± 0.3 22 ± 0.3
Pseudomonas aeruginosa 16 ± 0.3 17 ± 0.3 18 ± 0.3 19 ± 0.3 19 ± 0.3 20 ± 0.3 25 ± 0.3
Bacillus cereus 5 ± 0.3 6 ± 0.3 6.5 ± 0.3 8 ± 0.3 9 ± 0.3 11 ± 0.3 22 ± 0.3

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2.4. Antimicrobial activity around 5 µl ± 2 µl approximately. Not only these the action of the
cinnamon loaded silver nanoparticles can be seen best on gram negative
In case of disc diffusion method Muller Hinton Agar media is pre- species of the bacteria as we can see on Escherichia coli, Pseudomonas
pared by dissolving as per manufacturing company. Pathogens used in aeruginosa and it can be feebly seen in gram positive bacteria like
these studies are: Staphylococcus aureus, Escherichia coli, Bacillus cereus Bacillus cereus and Staphylococcus aureus (Figs. 7 and 8, Graphs 1 and 2
and Pseudomonas species which were from University of Madras, and Table 2).
PGIBMS. Inoculate the pathogens into the peptone water and incubate
it for 2–3 h at 37 °C. After sterilization, let the media to become cool and 4. Discussion
pour 15–17 ml the Muller Hinton Agar in petri plates under the sterile
conditions (laminar flow). Each plate was inoculated by fresh patho- Silver nanoparticles were synthesized from cinnamon extract bio-
gens. Zone of inhibition was measured to determine the efficiency of synthetically. Powdered cinnamon was taken 1 g and dissolved in 25 ml
the synthesized silver nanoparticles and their comparative analysis of distilled water. After continuous mixing and heating the solution at
(Premkumar et al., 2017, Sudhakar et al., 2017). 60–70 °C for 25–30 min. Then the solution is filtered and the filtrate
hence obtained is followed by the synthesis of silver nanoparticles.
3. Results Taking 0.5:4.5 dilution, mixing 10 µl of AgNO3, the resultant solution is
kept for incubation in dark room for 48 h for reduction (Nanda and
3.1. Synthesis of silver nanoparticles Majeed, 2014; Nanda and Raghavan, 2014). Finally the obtained silver
nanoparticles are synthesized for characterization such as UV–vis
3.1.1. UV–vis spectroscopy analysis spectroscopy, EDAX, FTIR and FESEM.
The silver nanoparticles obtained from the cinnamon sample are in UV visible spectroscopy is primary confirmation of the presence of
liquid medium i.e. aqueous solution. From visual analysis they exhibit silver nanoparticles (Nanda and Saravanan, 2013). The absorbance
the reddish brown color in the aqueous solution. The reason for such peaks came at 410–425 nm and for further confirmation FTIR is used to
color is the reduction of AgNO3 to Ag ions. It is generally recognized identify the possible biomolecules responsible for the reduction of the
that UV–Vis spectroscopy could be used to examine the size and shape Ag+ ions and escaping of the bio reduced silver nanoparticles synthe-
controlled nanoparticles in aqueous suspensions (Ranjit and Baquee, sized using Cinnamon extract.
2013). After the incubation of cinnamon for 48 h the UV–Vis spectro- FESEM revealed a uniform alignment of silver nanoparticles having
scopy shows the result of 412 nm. Then the produced silver nano par- size in the range of 50–70 nm and mono dispersed, spherical in shape,
ticles are subjected to the stability test for 3 days. Then after 3 days the whereas Kavitha et al. (2013) synthesized silver nanoparticles using
UV-Vis spectroscopy, the result shown as 423.2 nm (Fig. 3). Cinnamomum and obtained particle size in the range of 100 ± 25 nm.
But in our studies particle obtained in the size range 50–70 nm. XRD
3.1.2. FTIR (Fourier transform of Infrared radiations) pattern showed a peaks indicating the crystalline nature of the reduced
To identify the possible biomolecules responsible for the reduction silver nanoparticles 27.743°, 32.161°, 38.025°, 46.159°and 54.720°; this
of the Ag+ ions and escaping of the bio reduced silver nanoparticles was correlating with the work of Anuradha et al. (2010). Thus further
synthesized using cinnamon extract (Fig. 4 and Table 1). studies can be carried out to reduce the particle size thereby increasing
the anti-bacterial effect.
3.1.3. FESEM (Field emission scanning electron microscope) In the current research work silver nanoparticles were synthesized
The FESEM analysis of the cinnamon silver nanoparticles clearly by Cinnamon extract. The synthesized silver nanoparticles was sub-
shows that the particles size lies between the 50 and 70 nm in diameter jected to UV–vis spectrometer to determine the absorption range which
(Fig. 5). was found to be around 410–425 nm and confirmed the presence of
silver nanoparticles (Nanda and Saravanan, 2009). Further to confirm
3.1.4. X-ray diffraction analysis the presence of silver nanoparticles the synthesized particles was con-
Structural characterization has been found using XRD analysis and firmed by various analytical techniques such as FTIR, XRD and FESEM.
the typical XRD pattern for sample cinnamon silver nanoparticles was FTIR study revealed the presence of silver nanoparticles followed by
shown in the figure. X-ray diffract gram of the silver nanoparticles XRD. The size of synthesized silver nanoparticles was determined by
showed 5 distinct diffraction peaks at 27.743°, 32.161°, 38.025°, FESEM method the size was around 50–70 nm (Lavanya et al., 2013).
46.159°and 54.720°with inter planer spacing (d) of 3.21, 2.78, 2.36, All these studies revealed that the characteristic of synthesized nano-
1.96 and 1.67 respectively (Fig. 6). particles from extract of Cinnamon.
To determine the efficacy of the synthesized silver nanoparticles
3.2. Anti-bacterial activity antimicrobial activity of silver nanoparticles was determined by Kirby
Bauer method against some positive and negative bacteria (Lin et al.,
With obtained results and plotted bar diagram we can observe the 2011). In the present study synergistic activity showed efficient anti-
inhibition zones for various concentration of silver nanoparticles 10 µl, bacterial activity against the test pathogens when compared with the
20 µl, 30 µl, 40 µl, 50 µl and 60 µl respectively. So based on antibacterial nanoparticles alone. So, further studies can be done to investigate that
activities by different concentration of cinnamon silver nanoparticles the combined extract and silver nanoparticles with antibiotic can be
(Lee, 2007) we can conclude that as the concentration of the nano commercially used in pharmaceutical industry for much drug resistant
particles ascends the antibacterial activities also goes up with it. Since pathogens after determining their toxicity levels. So it cannot be only
we have carried out till 60 µl of concentration, The best zone of in- used as anti-bacterial but also antiviral, antifungal, and anticancer
hibition we have obtained for Staphylococcus aureus, Escherichia coli, agent (Manjumeena et al., 2014).
Pseudomonas aeruginosa and Bacillus cereus are 19 mm, 13 mm, 20 mm
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