Plant-mediated synthesis of nanoparticles has been an emerging research area worldwide for its inherent features such as rapidity, simplicity, environment-friendly, and cost-effectiveness. The rapid and simple approach was used for the synthesis of silver nanoparticles (AgNPs) (AgNO3) using stem of Berberis aristata (Daruharidra) by biologically reducing AgNO3 with the aqueous extract of plant. The formation of AgNPs was indicated by the color change from slightly yellowish to yellowishbrown. Various techniques were used to characterize biosynthesized NPs by ultraviolet-visible, Fourier transform infrared, scanning electron microscopy, and transmission electron microscopy analysis. The biosynthesized AgNPs were also tested for antimicrobial activity against pathogenic bacteria such as Escherichia coli and Pseudomonas aeruginosa.

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Green synthesis of silver nanoparticles using stem extract of Berberis aristata
and to study characterization and its antimicrobial activity.
Article in Journal of Pharmacy Research · May 2018
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2. Journal of Pharmacy Research | Vol 12 • Issue 6 • 2018840
INTRODUCTION
Much excitement has been created among researchers
at the birth of matter at the nanoscale which has
promised to metamorphose many aspects of material
sciences. The field of nanotechnology is related
with the evolution of exploratory processes for
the production of nanoparticles of various sizes,
controlled disparity, and shapes.[1]
Silver being
important metal, silver nanoparticles (AgNPs) have
a number of applications from electronics to medical
diagnosis.[2]
The various techniques available for the
preparation of AgNPs are electrochemical method,[3]
chemical reduction method,[4]
and microbial synthesis
method.[5]
Methods other than biological one involve
the use of chemicals and high energy requirements
which are rather difficult.[6]
Hence, it is necessary to
synthesize AgNPs by a route which is cost effective
and eco-friendly.
The AgNPs can be synthesized using microorganisms,
plants, and viruses or their by-product such as proteins
and lipids. The synthesis rate of AgNPs is much
faster in plant-mediated synthesis rather than using
microorganisms.[6]
With the huge plant diversity,
much more plants are still not explored for the
synthesis of AgNPs. The extracts of Tabernaemontana
divaricata,[7]
Chrysanthemum indicum,[8]
Azadirachta
indica,[6]
Calliandra haematocephala,[9]
and Pleurotus
citrinopileatus[10]
plants reported as an alternate source
for the synthesis of AgNPs.
In Ayurveda, Berberies aristata (B. aristata) is a
versatile medicinal plant used singly or in combination
with other medicinal plants for treating a variety of
ailments such as enlargement of spleen, leprosy,
rheumatism, fever, morning or evening sickness,
and snakebite.[11-14]
Stem extract of B. aristata (Daru
Haldi or Daruharidra) found in the Himalayas in
India till date has not been used for nanoparticle
synthesis. Daruharidra is used for the treatment of
skin disorders, internal medication as well as external
applications.[15]
It is used in inflammation, wound
healing, skin diseases, and jaundice.[16]
Therefore, the
plant is used as reducing agent for converting silver
ions into AgNPs. AgNPs have been extensively used
as antibacterial agents in the health industry, food
storage,[17]
nanoscale sensors,[18]
larvicidal activity,[19]
and anticancer activity.[8,20]
Green synthesis of silver nanoparticles using stem extract
of Berberis aristata and to study its characterization and
antimicrobial activity
Sahiba Kaur Saddal, Tejaswini Telang, Vivek P. Bhange, A. P. Kopulwar, S. R. Santra, Manju Soni*
Research Article
ABSTRACT
Plant-mediated synthesis of nanoparticles has been an emerging research area worldwide for its inherent features such as
rapidity, simplicity, environment-friendly, and cost-effectiveness. The rapid and simple approach was used for the synthesis
of silver nanoparticles (AgNPs) (AgNO3
) using stem of Berberis aristata (Daruharidra) by biologically reducing AgNO3
with
the aqueous extract of plant. The formation of AgNPs was indicated by the color change from slightly yellowish to yellowish-
brown. Various techniques were used to characterize biosynthesized NPs by ultraviolet-visible, Fourier transform infrared,
scanning electron microscopy, and transmission electron microscopy analysis. The biosynthesized AgNPs were also tested
for antimicrobial activity against pathogenic bacteria such as Escherichia coli and Pseudomonas aeruginosa.
KEY WORDS: Berberies aristata, Eco-friendly, Fourier transform infrared, Green synthesis, Nanoparticles, Scanning
electron microscopy, Transmission electron microscopy, Ultraviolet-visible
Department of Biotechnology, Priyadarshini Institute of Engineering and Technology, Nagpur, Maharashtra, India
*Corresponding author: Manju Soni, Department of Biotechnology, Priyadarshini Institute of Engineering and
Technology, Nagpur - 444 019, Maharashtra, India. E-mail: manjusonipiet@gmail.com
Received on: 05-02-2017; Revised on: 19-03-2018; Accepted on: 04-05-2018
Access this article online
Website: jprsolutions.info ISSN: 0974-6943

3. Sahiba Kaur Saddal, et al.
Journal of Pharmacy Research | Vol 12 • Issue 5 • 2018 841
MATERIALS AND METHODS
Preparation of Plant Extract
The stem of Daruharidra was brought from Wagh
Brother’s Agency, Itwari region, Nagpur. These parts
of plants were washed with distilled water to remove
debris and other contamination and dried in hot air
oven at 60°C for 24 h. Dried parts of plant were then
crushed into fine powder using domestic blender.
Then, 10 g of plant powder was weighed individually,
kept in a beaker containing 100 ml of distilled water,
and boiled at 60°C for 20 min. The extracts were
filtered using Whatman filter paper no.1 and stored in
a refrigerator for further use.
Synthesis of AgNPs
Aqueous solution of 1 mM of AgNO3
was prepared.
5 ml of prepared extract Daruharidra was added in
flask containing 30 ml of 1 mM AgNO3
solution.
The solution was then kept in a light as well as dark
condition for 24 h incubation. The color change
was observed from light yellow to dark brown after
incubation, which indicates the bioreduction of
Ag+
ions.
Characterization of Synthesized AgNPs
Ultraviolet-visible (UV-VIS) spectrometry
Optical property of AgNPs was determined by UV-
VIS spectrophotometer. The reduction of Ag+
ions was
observed by measuring spectrum in the range of 200–
800 nm by comparing with control (distilled water +
plant extract) as blank.
Fourier transform infrared (FTIR) analysis
The chemical composition of the synthesized AgNPs
was studied using FTIR spectrometer. It is used to
analyze possible biomolecules and also bonding
interactions. The absorbance was measured in the
range of 400–4000 nm. Sample was placed into one
of the KBr plates, and the plates were kept into the
sample holder.
Scanning electron microscopy (SEM)
SEM is the type of electron microscope that produces
an image of a sample by scanning it with a focused
beam of electrons. The electrons interact with atoms
in a sample, producing various signals that contain
information about the sample’s surface topography and
composition. SEM can achieve resolution better than
1 nm. Specimens can be observed in a high vacuum, wet
condition, and a wide range of cryogenic temperatures.
Samples for SEM can be solid, bulk specimens of any
size that will fit the specimen chamber.
Transmission electron microscopy (TEM)
TEM is a microscopy technique in which a beam of
electrons is transmitted through an ultrathin specimen,
interacting with the specimen as it passes through it.
A thin specimen is exposed to high-energy electron
beam. It gives microstructure and nanostructure size
and morphology.
Antimicrobial Assay
Antimicrobial analysis was done on the Gram-
negative bacteria (Escherichia coli and Pseudomonas
aeruginosa) by well-diffusion method using pour
plate technique. Nutrient broth cultures were prepared
using E. coli and P. aeruginosa and incubated at 37°C
for 24 h (Orbital Shaking Incubator, BIO-TECHNICS
INDIA). 1 ml of bacterial culture was inoculated in the
nutrient agar medium, and the plates were made using
pour plate method and allowed for solidification.After
solidification, wells were punched using sterile tips.
200 µl of AgNPs were loaded and incubated at 37°C
for 48 h (Cooling Incubator, REMI). Antimicrobial
activity was measured based on the zone of inhibition
obtained around the wells.
RESULTS AND DISCUSSION
UV-VIS Spectrometry
Reduction of silver ions into AgNPs during exposure
to plant extracts was observed as a result of the change
in color, which is the primary method to confirm the
synthesis ofAgNPs [Figure 1].[21]
The sharp absorption
of AgNPs was observed at 481 nm (1.9187 ABS)
for B. aristata (Daruharidra) stem [Figure 2]. The
peak was raised due to the effect of surface palsmon
resonance surface plasmon resonance of electrons in
the reaction mixture,[22]
which is also responsible for
the brown color.[23]
FTIR Analysis
The FTIR spectrum of synthesized AgNPs from
Daruharidra is shown in Figure 3 with four main
bands. The broadband appearing at 3450/cm is signed
for O-H stretching vibration, indicating the presence
of hydroxyl groups. The peaks at 1600/cm correspond
to C-C and C-N stretch. The peaks at 1400/cm
correspond to C=C and phenolic hydroxyl vibrations.
Figure 1: Synthesized silver nanoparticles from Daruharidra

4. Sahiba Kaur Saddal, et al.
Journal of Pharmacy Research | Vol 12 • Issue 6 • 2018842
The peaks at 1000/cm correspond to C-O vibrations.
These functional groups have been reported to play
a crucial role in stability, capping of AgNPs.[24-26]
.
From FTIR results, it can be concluded that some of
the bioorganic compounds from B. aristata extract
formed the strong coating or capping on the AgNPs.
SEM Analysis
The SEM images obtained at 10 µm, 5 µm, and 1 µm
are shown in Figure 4. The SEM image showed the
presence of high-density and compact agglomerates
of silver particles. The nature of AgNPs was found
to be highly granulated. The images also showed
the presence of various shapes such as spherical,
ellipsoidal, and few irregular fused agglomerates.
Similar kind of result was also obtained with Coffea
arabica seed.[25]
TEM Analysis
TEM has been used to identify the size, shape, and
morphology of nanoparticles. TEM micrograph of
the synthesized AgNPs at 100, 50, and 20 nm scale
Figure 2: Ultraviolet-visible spectra of silver nanoparticles
synthesized from Daruharidra
Figure 3: Fourier transform infrared spectra of silver
nanoparticles synthesized from Daruharidra
is shown in Figure 5. It was found that AgNPs were
spherical in shape and SAD pattern showed definite
crystalline ring structure. Similar kind of result was
reported with Pleurotus.[10]
Antimicrobial Activity
The results of antimicrobial activity of synthesized
AgNPs by well diffusion method are shown in
Table 1. Silver is a naturally occurring element which
is non-toxic and does not accumulate in the body to
cause harm effects and is considered to be safe for
the environment. Most manufactured goods such as
washing machines, air conditioners, and refrigerators
are using linings of AgNPs for their antimicrobial
qualities.[24]
We observed that the synthesized AgNPs
have antimicrobial activity against the Gram-negative
bacteria E. coli and P. aeruginosa [Figure 6 and 7].
Figure 4: Scanning electron microscopy images of silver
nanoparticles by Daruharidra (a) 10 µm, (b) 5 µm, and
(c) 1 µm
a b
c
Figure 5:Transmission electron microscopy images of silver
nanoparticles from Daruharidra at (a) 100 nm, (b) 50 nm, (c)
20 nm, and (d) 10 nm
a b
c d

5. Sahiba Kaur Saddal, et al.
Journal of Pharmacy Research | Vol 12 • Issue 5 • 2018 843
AgNPs were reported to be more efficient than silver
ions in terms of its antimicrobial activity.[26]
CONCLUSION
The present study is aimed to develop eco-friendly,
fast, and cost-effective method for the biosynthesis of
AgNPs from B. aristata. The silver ions are exposed
to an aqueous solution of Daruharidra stem, and
a rapid color change of plant extract signifies the
biosynthesis of AgNPs, which was confirmed by UV
spectroscopic method. This eco-friendly method could
be competitive alternative to the conventional physical
and chemical method used for the synthesis of silver
nanoparticle and has a potential to use in biomedical
applications. These green-synthesized AgNPs are
polydispersed, spherical with agglomeration and
Table 1: Zone of inhibition obtained by well diffusion
method
Samples Microorganism Zone of
inhibition (mm)
Daruharidra–
AgNPs
E. coli 7
P. aeruginosa 7
AgNPs: Silver nanoparticles, E. coli: Escherichia coli, P. aeruginosa:
Pseudomonas aeruginosa
Figure 7: Antimicrobial activity of silver nanoparticles by
Daruharidra against E. coli.
have sizes ranging from 20 to 100 um which shows
antimicrobial activity against Gram-negative bacteria
E. coli and P. aeruginosa and acts as a potential
antimicrobial agent. Moreover, high amount of small-
sized nanoparticles can be produced with a little
amount of plant extract and is beneficial for the society
and environment. However, further investigation is
needed to identify the scaling up of this extract of the
biosynthesis of AgNPs for its medicinal use.
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