Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 134 (2015) 326–332

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Spectrochimica Acta Part A: Molecular and

Biomolecular Spectroscopy
journal homepage: www.elsevier.com/locate/saa

Facile green synthesis of silver nanoparticles using seed aqueous extract

of Pistacia atlantica and its antibacterial activity
Babak Sadeghi a,⇑, Amir Rostami b, S.S. Momeni b
a
Department of Chemistry, Guilan Science and Research Branch, Islamic Azad University, Guilan, Iran
b
Department of Chemistry, Tonekabon Branch, Islamic Azad University, Tonekabon, Iran

h i g h l i g h t s g r a p h i c a l a b s t r a c t

 Biosynthesis of Ag-NPs has been done ‘‘We describe the synthesis of silver nanoparticles (Ag-NPs) using seed aqueous extract of Pistacia
with Pistacia atlantica (PA) extract. atlantica (PA) and its antibacterial activity. UV–visible spectroscopy, X-ray diffraction (XRD), Fourier
 It was observed that the growths of transform infra red spectroscopy (FTIR), transmission electron microscopy (TEM), scanning electron
Ag-NPs are stopped within 35 min. microscopy (SEM), and X-ray energy dispersive spectrophotometer (EDAX) were performed to ascertain
 SEM indicated that, S. aureus was the formation of Ag-NPs. It was observed that the growths of Ag-NPs are stopped within 35 min of
disappeared by addition of Ag-NPs. reaction time. The inhibition was observed in the Ag-NPs against S. aureus. The results suggest that the
synthesized Ag-NPs act as an effective antibacterial agent. It is confirmed that Ag-NPs are capable of ren-
dering high antibacterial efficacy and hence has a great potential in the preparation of drugs used against
bacterial diseases. The scanning electron microscopy (SEM(, indicated that, the most strains of S. aureus
was damaged and extensively disappeared by addition of Ag-NPs. The results confirmed that the (PA) is a
very good eco friendly and nontoxic source for the synthesis of Ag-NPs as compared to the conventional
chemical/physical methods.’’

a r t i c l e i n f o

Article history:
Keywords
Received 2 April 2014
Received in revised form 15 May 2014
Accepted 28 May 2014 Introduction: X-ray energy dispersive spectrophotometer (EDAX) methods [1–5]. Silver
were performed to nanoparticle
Available online 19 June 2014 ascertain the formation of Ag-NPs. It was observed twothat the growths
decades due of
to Ag-
their wide app
NPs are stopped within 35 min of reaction time.investigations
The synthesized ofAg-NPs
the use of plants
tle>In the present work, we describe were characterized by a peak at 446 nm in the the reductant for carrying out the s
Nanotechnology is a broad interdisciplinary
UV–visible areaconfirmed
of research, development and industrial
the synthesis of silver nanoparticles (Ag-NPs) using seed aqueous extract of Pistacia atlanticaspectrum. XRD
(PA) and its the crystalline
antibacterial activity. nature
UV–visible
phologies
of the
spectroscopy, X- including
in high yields,
activity which has been growing rapidly of
nanoparticles worldwide
27 nm for
size. the
The past
XRD decade.
peaks at Metallic
38°, 44°, 64° nanoparticles
and 77° canand
be
ray diffraction (XRD), Fourier transform infra red spectroscopy (FTIR), transmission electron microscopy (TEM), scanning electron microscopy (SEM),
of specific sizes and morphologies can be readily synthesized using [10]; but the
chemical anduse of surfactant in t
physical

⇑ Corresponding author. Tel.: +98 192 427 1105; fax: +98 192 427 4409.
E-mail addresses: [email protected], [email protected] (B. Sadeghi).

http://dx.doi.org/10.1016/j.saa.2014.05.078
1386-1425/Ó 2014 Elsevier B.V. All rights reserved.
 B. Sadeghi et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 134 (2015) 326–332 327

the methods
indexed to the (1 1 1), (2 0 0), (2 2 0) and (3 1 1) Bragg’s reflectionsreported in literature
of cubic structure are extremely
of metallic expensive
silver, respectively. Theand
FTIRthey also
result involve
clearly thethat
showed usetheof
extracts containing OH as a functional group act toxic, in capping the nanoparticles
hazardous chemicalssynthesis. Antibacterial
as the stabilizers whichactivities of Ag-NPs
may pose were environmental
potential tested against theandgrowth
bio-
of Gram-positive (S. aureus) using SEM. The inhibition was
logical observed
risks. in theofAg-NPs
Because against S. aureus.
the increasing The resultsconcerns
environmental suggest that
by the synthesized
chemical Ag-NPs
synthesis act as
routes,
an effective antibacterial agent. It is confirmed that Ag-NPs are capablesustainable
an environmentally of renderingsynthesis
high antibacterial
processefficacy
has led andtohence has a great
biomimetic potential in
approaches, the
which
preparation of used drugs against bacterial diseases. The scanning electron microscopy (SEM), indicated that, the most strains of S. aureus was damaged
refers to applying biological principles in materials formation. Bio-reduction is one of the fun-
and extensively disappeared by addition of Ag-NPs. The results confirmed that the (PA) is a very good eco friendly and nontoxic source for the synthesis of
damental processes in the biomimetic synthesis. The stability, shape, size, and morphologies of
Ag-NPs as compared to the conventional chemical/physical methods.
metal nanoparticles strongly depend on the method of preparation, type, nature of reductants,
and concentration of stabilizers (polymers, ligands, solid matrix and surfactants) [11]. The sur-
facework,
In the present plasmon resonance
we describe and large
the synthesis effective
of silver scattering
nanoparticles (Ag-NPs)cross
using section of individual
seed aqueous silveratlantica
extract of Pistacia nano-
particles
(PA) and its make
antibacterial them
activity. ideal spectroscopy,
UV–visible candidatesX-ray for diffraction
molecular labeling
(XRD), Fourier where
transformphenomena such as
infra red spectroscopy
surface enhanced
(FTIR), transmission Raman (TEM),
electron microscopy scattering
scanning(SERS) canmicroscopy
electron be exploited
(SEM),[12]. In addition,
and X-ray silver nanoparti-
energy dispersive
spectrophotometer (EDAX) were performed to ascertain the formation of Ag-NPs. It was observed that the growths of Ag-NPs are
cles play a significant role in the field of biology and medicine due to its attractive physiochem-
stopped within 35 min of reaction time. The synthesized Ag-NPs were characterized by a peak at 446 nm in the
ical properties. The strong toxicity of silver against wide range of microorganisms is well
UV–visible spectrum. XRD confirmed the crystalline nature of the nanoparticles of 27 nm size. The XRD peaks at 38°, 44°, 64° and
77° can beknown
indexed to and silver
the (1 1 1), (2nanoparticles
0 0), (2 2 0) and (3have beenreflections
1 1) Bragg’s recentlyofshown to beofametallic
cubic structure promising antimicrobial
silver, respectively. The
FTIR resultmaterial [13–17].
clearly showed that theSilver
extractsnanoparticles
containing OH ashave foundgroup
a functional to posses anti-inflammatory,
act in capping antiviral,
the nanoparticles synthesis.
Antibacterial activities of Ag-NPs and
anti-angiogenesis, were anti-platelet
tested against the growthand
activity of Gram-positive
cytotoxicity (S. against
aureus) using SEM.cells
cancer The inhibition
which makeswas
observed in the Ag-NPs against S. aureus. The results suggest that the synthesized Ag-NPs act as an effective antibacterial agent. It
them vital [18–20]. However, these methods employ toxic chemicals as reducing agents, or
is confirmed that Ag-NPs are capable of rendering high antibacterial efficacy and hence has a great potential in the preparation of
used drugsnonbiodegradable
against bacterial diseases.stabilizing
The scanning agents andmicroscopy
electron are therefore potentially
(SEM), indicated that, dangerous to of
the most strains the environ-
S. aureus was
damaged andment and biological
extensively disappeared systems
by addition [21]. Moreover,
of Ag-NPs. mostconfirmed
The results of thesethat methods
the (PA) isentail intricate
a very good controls
eco friendly and
or nonstandard.
nontoxic source Weofhave
for the synthesis Ag-NPs recently developed
as compared a reduction
to the conventional method of converting
chemical/physical methods. Ag nanospheres
into nanorods [22], nanoplates [23], their antibacterial activity [24,25], an improved an easy
synthetic route for silver nanoparticles in poly (diallyldimethylammonium chloride) (PDDA)
[26], synthesis of gold/HPC hybrid nanocomposite [27], preparation of ZnO/Ag nanocomposite
[28] and comparison nanosilver particles and nanosilver plates for the oxidation of ascorbic
acid [29]. Regarding the role of green chemistry, it was successfully demonstrated that size,
shape and the antibacterial activity silver nanoparticles by the reduction of Ag+ ions with
bio-reductants (Pistacia atlantica) largely depend on the nature of reducing agents, concentra-
tion and time of mixing of the reactants [30]. The methodology employed here is very simple,
easy to perform, inexpensive, and eco-friendly. Moreover, most of these methods entail intri-
cate controls or nonstandard. (Scheme 1).

Fig. 1. UV–vis spectra of an aqueous solution of Pistacia atlantica (PA) leaf extract in
presence of Ag+ ions at 30 °C. Reaction conditions: [Ag+] = 10.0  10 4 mol dm 3.

Scheme 1. Synthesis and characterization of stabilizes Ag nanoparticles with P. atlantica (PA) leaf extract reduces.
328 B. Sadeghi et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 134 (2015) 326–332

Fig. 2. FTIR spectra of P. atlantica (PA) powder before (A) and after (B) reaction with AgNO3.

Materials and methods For the antimicrobial activity meas

medium (tryptone 1.5%, yeast extr
Materials
Synthesis and characterization of sil
Silver nitrate (AgNO3) was obtained from Loba Chemie, India and used as received. All other
reagents used in the reaction were of analytical grade with maximum purity.
In a P. atlantica
typical (PA) procedure
reaction
leaves were collected from South of IRAN, and were cleaned with doubledry distilled water
leaf powder and25 mL of dis
with
shade-dried for a week at room temperature and further (PA) leaves were groundmethanol
(minimum to powder was added in
and stored for further study. For this experiment, nanoparticles have concentrations rangingshaker for 1
was placed in orbital
from 0.0976 to100 lg/mL. S. aureus) ATCC 51153) was used as a Gram-positive
nanoparticles bacterium.
various concentratio
was optimized to 1 mL. Further, 1
 B. Sadeghi et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 134 (2015) 326–332 329

Fig. 3. TEM images indicating the presence of spherical silver nanoparticles recorded at various magnifications (A and B).

which were reported

(AgNO3) solution and the solution was placed in orbital shaker at room temperature, for reduc-earlier in
tion of Ag+ to Ag0. The broth containing Ag-NPs was centrifuged at 10,000 bancerpmwasfor extremely
15 min, fol-sensitive t
lowing which the pellet was re-dispersed in the sterile distilled water interto particle.
get rid of any
uncoordinated biological molecules. The color change involved in the formation of silver nano-
particles. The purified pellets were then kept into petri plates and left inFTIR
the studies
oven for drying at
60 °C for 24 h. The colorless AgNO3 solution turned yellow to brown or reddish yellow to deep
red, indicated the formation of Ag-NPs. The dried Ag-NPs were scrappedFTIR out for the further were ca
measurements
study. reduction, capping and efficient
in proteins and have high availa
Antimicrobial activity studies of the control extract and (PA) an
seen that, in contrast to the con
S. aureus (ATCC 51153) was used as a Gram-positive bacterium. For the antimicrobial
changes activ-
in their respective vibr
ity measurement, bacteria cultures were incubated at 38 °C in Luria medium (tryptone 1.5%,
The interaction of Ag nanop
yeast extract 0.75%, sodium chloride 1.2%, agar 1%, Difco). 2935, 1629, 1515, 1384, 1156, 1
Antimicrobial activities of silver nanoparticles (Ag-NPs) have been tribution
investigated were against
confirmedS. with F
aureus as the model Gram-positive bacteria. The in vitro anti-bacterial activities
strong bandsof silver
were nano-
observed at 1
particles were examined according Melaiye and Feng [31–33]. The following microorganism
be identified that the changes
was used: Gram-positive Staphylococcus aureus. appeared at 3421 cm 1 in raw
narrower and shifted to 3391 c
Characterization of silver nanoparticles (Ag-NPs) reduced after encapsulation of
to C–N stretching of amine grou
The biosynthesis of the Ag-NPs in a solution was monitored by measuring but afterthe encapsulation
UV–vis spec- of nano
tra of the solutions (1:4 diluted) of the reaction mixture. UV–vis spectra –COOH group in
were recorded on the
dou-compoun
ble beam spectrophotometer (Shimadzu, model UV-1800, Kyoto, Japan)change from 300 in the spectra.
to 800 nm at1702 cm
a resolution of 1 nm. The distilled water was used as a blank. The Ag-NPs is masked
synthesized or disappeared.
with 8%
leaf extracts and 6 mM AgNO3 solution were characterized with the help of The absorption
scanning peak at 3421
electron
microscopy (model LEO 440i) equipped with X- ray energy dispersiveing vibration, get
spectrometer wider and sh
(EDAX).
Transmission electron microscopy (TEM) selected area electron diffraction from protein
(SAED) present
images werein the con
taken on Zeiss – EM10C – 80 kV operated at accelerating voltages of broader
40 and in 200
the kV.
spectrum
The of stab
observed reflection planes corresponding to fcc Ag-NPs (27 nm) in XRD are characteristic
diffraction patternof amide I whe
tively [8,37–39].
(Seisert Argon 3003 PTC using nickel filtered XD-3 Cu Ka radiations (k = 1.5418 In Fig. 2B, the
Å)). The purified
powders of Ag-NPs were subjected to FTIR spectroscopy (FTIR; Bomem 1629 cm 1.These
MB100). Thesemea-structural ch
surements were carried out on a Perkin–Elmer Spectrum-One instrument nanoparticles
in the diffuse proceed
reflec-via the c
tance mode at a resolution of 4 cm 1 in KBr pellets. For comparison, a drop FTIR ofstudies
8% leaf have confirmed the
extracts
was mixed with KBr powder and pelletized after drying properly. The ity to bind
pellets weremetal
later indicating
sub- tha
jected to FTIR spectroscopy measurement. nanoparticles (i.e., capping of si
bilize the medium. Comparison
Results and discussion Ag-NPs reveal only minor chang
tion bands; wave numbers vary
UV–vis spectral studies
TEM
UV–vis spectroscopy was ascertained to check the formation and stability of Ag-NPs in
The morphology
aqueous solution (Fig. 1.). The colorless AgNO3 solution turned yellow to brown and size of
or reddish yel-
images
low to deep red, indicated the formation of Ag-NPs. The appearance of the brownand they
color aredue
was shown in Fi
nanospherical
to the excitation of the surface plasmon resonance (SPR), typical of Ag-NPs having kmaxformed
valueswhere sh
330 B. Sadeghi et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 134 (2015) 326–332

Fig. 4. (A) SEM and (B) EDAX images showing the presence of silver nanoparticles and bioorganic components of Pistacia atlantica (PA).

in the range of 10–50 nm in size with 36 nm average size. The particles were monodisperse,
04-0784 revealing that synthesize
with only a few particles of different size. (Fig. 5) of Ag-NPs obtained were si
the Ag-NPs formed were calculate
SEM were good in agreement with TEM

SEM micro-graphs show aggregates of silver nanoparticles and the particles are inactivity
Antibacterial the range
of silver nanop
of 25–40 nm and there are not in direct contact even within the aggregates indicating the sta-
bilization of nanoparticles by capping agents (Fig. 4A). In EDAX strong signals wereinvestigated
We have observed the use o
from the silver atoms in the nanoparticles and weaker signals for carbon, oxygen,
bacterial potassium
agents. Such (PA) mediat
and chloride were provenients from biomolecules of (PA) (Fig. 4B). crobial activity towards test bacte
were observed with the S. aureus.
XRD colonization. This is consistent w
nanoparticles biosynthesized [40],
The crystalline nature of Ag-NPs was carried out using XRD where ent threestudy, the nanoparticles
diffraction peaks thus
were observed in the 2h range of 30–80°, which can be indexed asagents. (1 1 1),The
(2 0inhibition
0), (2 2 0),was observ
theJoint
(3 1 1) reflections of fcc structure metallic silver respectively similar to synthesized
Committee Ag-NPs
on act as an e
Powder Diffraction Standards (JCPDS) File No:
 B. Sadeghi et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 134 (2015) 326–332 331

Fig. 7. Zeta potential at different pH.

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Fig. 6. Representative SEM images showing reduced S. aureus colonization on Ag-Nps/extract (B) compared to extract (A). Arrows show bacteria.
332 B. Sadeghi et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 134 (2015) 326–332