Int. J. Nano Dimens.

5(6): 575-581 (Special Issue) 2014

ISSN: 2008-8868

Contents list available at IJND

International Journal of Nano Dimension
Journal homepage: www.IJND.ir

Green synthesis of silver nanoparticles using seed aqueous

extract of Olea europaea

ABSTRACT

I D
S
B. Sadeghi* Biosynthesis of silver nanoparticles with small size and
biostability is very important and used in various biomedical
Department of Chemistry, applications. In the present work, we describe the synthesis of silver
Tonekabon Branch, Islamic Azad
University, Tonekabon, Iran.
Received 08 November 2013
Accepted 29 January 2014

o f
nanoparticles (Ag-NPs) using seed aqueous extract of Olea europaea
(Oe) and its antibacterial activity. UV–visible spectroscopy, X-ray
diffraction (XRD), transmission electron microscopy (TEM), scanning
electron microscopy (SEM), and X-ray energy dispersive
spectrophotometer (EDAX) were performed to ascertain the formation of

e
Ag-NPs. It was observed that the growths of Ag-NPs are stopped within
35 min of reaction time. The synthesized Ag-NPs were characterized by a

v
peak at 449 nm in the UV–visible spectrum. XRD confirmed the

h i
crystalline nature of the nanoparticles of 34 nm size. The XRD peaks at
38, 44, 64 and 77 can be indexed to the (1 1 1), (2 0 0), (2 2 0) and (3
1 1) Bragg’s reflections of cubic structure of metallic silver, respectively.
The results confirmed that the (Oe) is a very good eco friendly and

c
nontoxic source for the synthesis of Ag-NPs as compared to the
conventional chemical/physical methods.

A r Keywords: Silver nanoparticles; Green chemistry; Scanning electron

microscopy (SEM); Olea europaea; X-ray diffraction (XRD).

INTRODUCTION

Utilizing plant extracts for the biosynthesis of silver

nanoparticles has gained importance in recent years due to the
enhancement of chemical, physical, biological and optoelectronic
* Corresponding author:
properties of the particles formed by this green process. Metal and
Babak Sadeghi semiconductor nanoparticles are very important due to their unusual
Department of Chemistry, size and shape dependant properties. Nanotechnology is a broad
Tonekabon Branch, Islamic Azad interdisciplinary area of research, development and industrial activity
University, Tonekabon, Iran. which has been growing rapidly worldwide for the past decade.
Tel +98 192 4271105
Metallic nanoparticles of specific sizes and morphologies can be
Fax +98 192 4272484
Email [email protected] readily synthesized using chemical and physical methods [1–5].

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Int. J. Nano Dimens. 5(6): 575-581 (Special Issue) 2014 Sadeghi

Silver nanoparticles have attracted the nanospheres into nanorods [22], nanoplates [23],
attention of the researchers in the last two decades their antibacterial activity [24, 25], an improved an
due to their wide applications in various fields. The easy synthetic route for silver nanoparticles in poly
literature is replete with the investigations of the (diallyldimethylammonium chloride( )PDDA) [26],
use of plants extracts [6], fungi [7], algae [8], synthesis of Gold/HPC hybrid nanocomposite [27],
proteins and enzymes [9] as the reductant for Preparation of ZnO/Ag nanocomposite [28] and
carrying out the syntheses of nanoparticles with a comparison nanosilver particles and nanosilver
variety of shapes and morphologies in high yields, plates for the oxidation of ascorbic acid [29].
including multi-branched advanced silver and/or Regarding the role of green chemistry, we have
gold nanomaterials [10]; but the use of surfactant in successfully demonstrated that size, shape and the
the green synthesis of silver sol has been neglected. antibacterial activity silver nanoparticles by the
Most of the methods reported in literature are reduction of Ag+ ions with bio-reductants (Olea
extremely expensive and they also involve the use europaea ) largely depend on the nature of reducing
of toxic, hazardous chemicals as the stabilizers agents, concentration and time of mixing of the
which may pose potential environmental and
biological risks. Because of the increasing
environmental concerns by chemical synthesis
D
reactants [30]. The methodology employed here is

I
very simple, easy to perform, inexpensive, and eco-
friendly.

S
routes, an environmentally sustainable synthesis
process has led to biomimetic approaches, which
refers to applying biological principles in materials EXPERIMENTAL
formation. Bio-reduction is one of the fundamental
processes in the biomimetic synthesis. The stability,
shape, size, and morphologies of metal
nanoparticles strongly depend on the method of
Materials

o f
Silver nitrate (AgNO3) was obtained from
Loba Chemie, India and used as received. All other

e
preparation, type, nature of reductants, and reagents used in the reaction were of analytical
concentration of stabilizers (polymers, ligands, grade with maximum purity. Olea europaea

v
solid matrix and surfactants) [11]. The surface (Oe)leaves were collected from South of IRAN,

i
plasmon resonance and large effective scattering and were cleaned with double distilled water and
cross section of individual silver nanoparticles shade-dried for a week at room temperature and

h
make them ideal candidates for molecular labeling further (Oe)leaves were ground to powder and
where phenomena such as surface enhanced Raman stored for further study. For this experiment,

c
scattering (SERS) can be exploited [12]. In nanoparticles have concentrations ranging from
addition, silver nanoparticles play a significant role 0.0976 to100 µg/mL.

A r
in the field of biology and medicine due to its
attractive physiochemical properties. The strong
toxicity of silver against wide range of
microorganisms is well known and silver
nanoparticles have been recently shown to be a
promising antimicrobial material [13–17]. Silver
Synthesis and characterization of silver
nanoparticles (Ag-NPs)
In a typical reaction procedure, Olea
europaea (Oe)leaf extract was prepared by taking 2
g of dry leaf powder with 25 mL of distilled water
nanoparticles have found to posses anti- in a conical flask along with 2 mL of methanol
inflammatory, antiviral, anti-angiogenesis, and anti- (minimum methanol was added in order to initiate
platelet activity and cytotoxicity against cancer the isolation of compounds). The extract was
cells which makes them vital [18–20]. However, placed in orbital shaker for 1 h and the extract was
these methods employ toxic chemicals as reducing filtered. For the synthesis of silver nanoparticles
agents, or nonbiodegradable stabilizing agents and various concentrations of leaf extracts were tried
are therefore potentially dangerous to the and then the extract to be used was optimized to 1
environment and biological systems [21]. mL. Further, 1 mL of the extract was added to 10
Moreover, most of these methods entail intricate mL of 1 mM silver nitrate (AgNO3) solution and
controls or nonstandard. We have recently the solution was placed in orbital shaker at room
developed a reduction method of converting Ag temperature, for reduction of Ag+ to Ag0. The broth

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Int. J. Nano Dimens. 5(6): 575-581 (Special Issue) 2014 Sadeghi

containing Ag-NPs was centrifuged at 10,000 rpm

for 15 min, following which the pellet was re-
dispersed in the sterile distilled water to get rid of
any uncoordinated biological molecules. The color
change involved in the formation of silver
nanoparticles. The purified pellets were then kept
into petri plates and left in the 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 scrapped out for the further study.

Characterization of silver nanoparticles (Ag-NPs)

Fig. 1. UV–vis spectra of an aqueous solution of Olea europaea (Oe)
The biosynthesis of the Ag-NPs in a leaf extract in presence of Ag+ ions at 32 °C. Reaction conditions:
solution was monitored by measuring the UV–vis
spectra of the solutions (1:4 diluted) of the reaction
mixture. UV–vis spectra were recorded on double
beam spectrophotometer (Shimazdu, model UV- TEM
I D
[Ag+] = 10.0 × 10−5 mol dm−3.

1800, Kyoto, Japan) from 300 to 800 nm at a

resolution of 1 nm. The distilled water was used as
a blank. The Ag-NPs synthesized with 8% leaf
extracts and 6 mM AgNO3 solution were
f S
The morphology and size of the
synthesized silver nanoparticles were determined
by TEM images and they are shown in Figure 2 (A
and B). The particles formed were spherical in
characterized with the help of scanning electron
microscopy (model LEO 440i) equipped with X-
ray energy dispersive spectrometer (EDAX)
(Bankar et al., 2010). Transmission electron
o
shape. The nanospherical formed where shown to
have high surface area. The nanoparticles formed
were in the range of 20–40 nm in size with 36 nm

e
average size. The particles were monodisperse,

v
microscopy (TEM) selected area electron with only a few particles of different size.

i
diffraction (SAED) images were taken on Zeiss -
EM10C - 80 KV operated at accelerating voltages

h
of 40 and 200 kV. The observed reflection planes
corresponding to fcc Ag-NPs (~27 nm) in XRD

c
diffraction pattern (Seisert Argon 3003 PTC using
nickel filtered XD-3Cu Ka radiations (k = 1.5418
A)).

RESULTS AND DISCUSSION

UV–vis spectral studies A r

UV–vis spectroscopy was ascertained to
check the formation and stability of Ag-NPs in
aqueous solution (Figure 1). The colorless AgNO3
solution turned yellow to brown or reddish yellow
to deep red, indicated the formation of Ag-NPs.
The appearance of the brown color was due to the
excitation of the surface plasmon resonance (SPR),
typical of Ag-NPs having λ max values which were
reported earlier in the visible range of 450–500 nm
[31-34] (Figure 1). The SPR absorbance was
extremely sensitive to the nature, size and shape of Fig. 2. TEM images indicating the presence of spherical silver
the particles formed, their inter particle. nanoparticles recorded at various magnifications (A and B).

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Int. J. Nano Dimens. 5(6): 575-581 (Special Issue) 2014 Sadeghi

SEM reported earlier [36-40] The particle size of the Ag-

SEM micro-graphs show aggregates of NPs formed were calculated using Debye–Scherrer
silver nanoparticles and the particles are in the equation which was around 38 nm, were good in
range of 15–30 nm and there are not in direct agreement with TEM results also.
contact even within the aggregates indicating the
stabilization of nanoparticles by capping agents
(Figure 3a). In EDAX strong signals were observed
from the silver atoms in the nanoparticles and
weaker signals for carbon, oxygen, potassium and
chloride were provenients from biomolecules of
(Oe) (Figure 3b) [35].

I D
f S
Fig. 4. XRD pattern of silver nanoparticles obtained using
Olea europaea (Oe).

e o
CONCLUSIONS

To conclude, the Ag-NPs were

produced by the use of the extract of Olea

i v europaea (Oe)as reducing and capping agent. In

this study, it was observed that the reaction is
rapid and is completed within 30 min at room

c h temperature. We have demonstrated an eco-

friendly, rapid green chemistry approach for the
synthesis of Ag-NPs by using (Qe), which pro-

r
vides a simple, cost effective and efficient way for
the synthesis of Ag-NPs. Therefore, this reaction
pathway satisfies all the conditions of a 100%

XRD
A
Fig. 3. (A) SEM and (B) EDAX images showing the presence of silver
nanoparticles and bioorganic components of Olea europaea (Oe).

The crystalline nature of Ag-NPs was

green chemical process. The amount of plant
material was found to play a critical role in
controlling the size and size dispersity of
nanoparticles in such a way that smaller silver
nanoparticles and narrow size distribution are
produced when more (Oe) extract is added in the
carried out using XRD where three diffraction reaction medium. The present study showed an
peaks were observed in the 2θ range of 30–80, innovative way for synthesizing antimicrobial Ag-
which can be indexed as (1 1 1), (2 0 0), (2 2 0), NPs using natural products which can be used in
(311) reflections of fcc structure metallic silver various biomedical applications. The results
respectively similar to Joint Committee on Powder confirmed that the (Oe) is a very good eco friendly
Diffraction Standards (JCPDS) file no: 04- 0784 and nontoxic source for the synthesis of Ag-NPs as
revealing that synthesized Ag-NPs are of pure compared to the conventional chemical/physical
crystalline silver. The XRD patterns in (Figure 4) methods.
of Ag-NPs obtained were similar to the results

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Int. J. Nano Dimens. 5(6): 575-581 (Special Issue) 2014 Sadeghi

ACKNOWLEDGEMENTS Mechanistic aspects of biosynthesis of

silver nanoparticles by several Fusarium
The financial and encouragement support oxysporum strains. J. Biomed. Nanotech. 3:
provided by the Research vice Presidency of 1-8.
Tonekabon Branch, Islamic Azad University, and
[8] Xie J., Lee J.Y., Wang D.I.C., Ting Y.P.,
Executive Director of Iran-Nanotechnology
Organization (Govt. of Iran). (2007), Identification of active
biomolecules in the high-yield synthesis of
single-crystalline gold nanoplates in algal
REFERENCES solutions. Small. 3: 672-682.
[9] Zhou Y., Chen W., Itoh H., Naka K., Ni
[1] Sun Y., Mayers B., Herricks T., Xia Y., Q., Yamane H., Chujo Y., (2001),
(2000), Polyol synthesis of uniform silver Preparation of a novel core-shell

D
nanowires: a plausible growth mechanism nanostructured gold colloid-silk fibroin

I
and the supporting evidence Nano Lett. 3: bioconjugate by the protein in situ redox
955–960. technique at room temperature. Chem.

S
[2] Srikanth H., Hajndl R., Chirinos C., Commun. 2518-2519.
Sanders J., (2001), Magnetic studies of [10] Zhai X., Efrima S., (1996), Silver Colloids

f
polymer-coated Fe nanoparticles and Macroemulsions of Metal Interfacial
synthesized by microwave plasma Colloidal Films:  Interaction with
polymerization. Appl. Phys. Lett. 79: 3503
– 3505.
[3] Valle-Orta M., Díaz D., Santiago P.,
Reguera E., Vázquez-Olmos A., (2008),
e o
[11]
Dithizone. J. Phys. Chem. 100: 10235-
10242.
El-Sayed M.A., (2001), Some Interesting
Properties of Metals Confined in Time and

iv
Instantaneous synthesis of stable zero- Nanometer Space of Different Shapes. Acc.
valent metal nanoparticles under standard Chem. Res. 34: 257-264.
[12] Sileikaite A., Prosycˇevas I., Puišo J.,

h
reaction conditions. J. Phys. Chem. B. 112:
14427-14434. Juraitis A., Guobiene A., (2006), Analysis

Iron

r
nanoparticles:
c
[4] Guo L., Huang Q., Li X., Yang S., (2001),
Synthesis
applications in surface enhanced Raman
and

[13]
of Silver Nanoparticles Produced by
Chemical Reduction of Silver Salt
Solution. Mater. Sci.12: 287–291.
Sondi I., Salopek-Sondi B., (2004), Silver

A
scattering and electrocatalysis. Phys. Chem.
Chem. Phys. 3: 1661-1665. nanoparticles as antimicrobial agent: a case
[5] Alqudami A., Annapoorni S., (2007), study of E. coli as a model for gram-
Fluorescence from metallic silver and iron negative bacteria. J. Colloid Interf. Sci.
nanoparticles prepared by exploding wire 275: 177–182.
technique. Plasmonics. 7: 5-13. [14] Sotiriou G.A., Pratsinis S.E., (2010),
[6] Shankar S.S., Rai A., Ahmad A., Sastry Antibacterial activity of nanosilver ions and
M., (2004), Rapid synthesis of Au, Ag, and particles. Sci. Technol. 44: 5649–5654.
bimetallic Au core Ag shell nanoparticles [15] Huang J., Zhan G., Zheng B., Sun D., Lu
using Neem (Azadirachta indica) leaf F., Lin Y., Chen H., Zheng Z., Zheng Y.,
broth. J. Colloid Interf. Sci. 275: 496– Li Q., (2011), Biogenic silver nanoparticles
502. by Cacumen Platycladi extract: synthesis,
[7] Duran N., Marcato P.D., De Souza G.I.H., formation mechanism, and antibacterial
Alves O.L., Esposito, E., (2009),

Submit your manuscript to www.ijnd.ir 579

www.SID.ir
Int. J. Nano Dimens. 5(6): 575-581 (Special Issue) 2014 Sadeghi

activity. Ind. Eng. Chem. Res. 50: 9095– Low-dimensional Systems and
9106. Nanostructures. 40: 3183-3186.
[16] He L., Gao S., Wu H., Liao X., He Q., Shi [23] Sadeghi B., Sadjadi M.A.S., Vahdati
B., (2012), Antibacterial activity of silver R.A.R., (2009), Nanoplates controlled
nanoparticles stabilized on tannin-grafted synthesis and catalytic activities of silver
collagen fiber. Mater. Sci. Eng. C. 32: nanocrystals. Superlatt. Microstruc. 46:
1050–1056. 858-863.
[17] Kaviya S., Santhanalakshmi J., [24] Sadeghi B., Jamali M., Kia Sh., Amini Nia
Viswanathan B., Muthumary J., Srinivasan A., Ghafari S., (2010), Synthesis and
K., (2011), Biosynthesis of silver characterization of silver nanoparticles for
nanoparticles using citrus sinensis peel antibacterial activity. Int. J. Nano Dimens.
extract and its antibacterial activity. 1: 119-124.

[18]
Spectrochim. Acta Part A. 79: 594–598.
Elumalai
Hemachandran
E.K.,
J.,
Prasad
Therasa
K.V.,
S.V.,
[25]

I D
Sadeghi, B., Garmaroudi F., Hashemi M.,
Nezhad H.R., Nasrollahi A., Ardalan
Sima., Ardalan Sahar., (2012), Comparison

S
Thirumalai T., David E., (2010), of the antibacterial activity on the
Extracellular synthesis of silver nanosilver shapes: nanoparticles, nanorods

f
nanoparticles using leaves of Euphorbia and Nanoplates. Adv. Powder Technol. 23:
hirta and their antibacterial activities. J. 22-26.

[19]
Pharm. Sci. Res. 2: 549–554.
Safaepour M., Shahverdi A.R., Shahverdi
H.R., Khorramizadeh M.R., Gohari A.R.,

e o
[26] Sadeghi B., Pourahmad A., (2012),
Synthesis of
(diallyldimethylammonium
hybrid nanocomposite. Adv. Powder
silver/poly
chloride)

v
(2009), Green Synthesis of Small Silver
Technol. 22: 669-673.

i
Nanoparticles Using Geraniol and Its
[27] Sadeghi B., Ghammamy Sh., Gholipour,
Cytotoxicity against Fibrosarcoma-Wehi
Z., Ghorchibeigy M., Amini Nia A.,

h
164. Avicenna J. Med. Biotechnol. 1: 111–
(2011), Gold/HPC hybrid nanocomposite
115.

c
constructed with more complete coverage
[20] Kalishwaralal K., Deepak V., Pandian

r
of gold nano-shell. Mic & Nano Lett. 6:
S.R.K., Kottaisamy M., Manikanth S.B.,
209-213.
Kartikeyan B., Gurunathan S., (2010),
[28] Sadeghi B., (2014), Preparation of ZnO/Ag

A
Biosynthesis of silver and gold
nanocomposite and coating on polymers for
nanoparticles using Brevibacterium casei.
anti-infection biomaterial application.
Colloid. Surface. B. 77: 257–262.
Spectrochimica Acta Part A: Molec.
[21] Nadagouda M.N., Hoag G., Collins J., Biomol. Spec. 118: 787-792.
Varma R.S., (2009), Green synthesis of Au [29] Sadeghi B., Meskinfam M., (2012), A
nanostructures at room temperature using direct Comparison of nanosilver particles
biodegradable plant surfactants. Cryst. and nanosilver plates for the oxidation of
Growth Des. 9: 4979–4983. ascorbic acid. Spectrochimica Acta Part A:
[22] Sadjadi M.A.S, Sadeghi B., Meskinfam M., Molec. Biomol. 97: 326-328.
Zare K., Azizian J., (2008), Synthesis and [30] Umesh B., Jagtap V., Bapat A., (2013),
characterization of Ag/PVA nanorods by Green synthesis of silver nanoparticles
chemical reduction method. Physica E: using Artocarpus heterophyllus Lam. seed

580 Submit your manuscript to www.ijnd.ir

www.SID.ir
Int. J. Nano Dimens. 5(6): 575-581 (Special Issue) 2014 Sadeghi

extract and its antibacterial activity. Ind. [37] He S., Zhang Y., Guo Z., Gu N., (2008),
Crops and Prod.46: 132– 137. Biological synthesis of gold nanowires
[31] Feng Q.L., Wu J., Chen G.Q., Cui F.Z., using extract of Rhodopseudomonas
Kim T.N., Kim J.O., (2000), A mechanistic capsulate. Biotechnol. Prog. 24: 476–480.
study of the antibacterial effect of silver [38] Sheny D.S., Philip D., Mathew J., (2012),
ions on Escherichia coli and Rapid green synthesis of palladium
Staphylococcus aureus, J. Biomed. Mater. nanoparticles using the dried leaf of
Res. 52: 662–668. Anacardium occidentale. Spectrochim.
[32] Melaiye A., Sun Z., Hindi K., Milsted A., Acta A. 91: 35-38.
Ely D., Reneker D.H., Tessier C.A., [39] Basavaraja S., Balaji S.D., Lagashetty A.,
Youngs W.J., (2005), Silver(I)_imidazole Rajasab A.H., Venkataraman A., (2008),
cyclophane gem-diol complexes Extracellular biosynthesis of silver

D
encapsulated by electrospun tecophilic nanoparticles using the fungus Fusarium

I
nanofibers: formation of nanosilver semitectum. Mater. Res. Bull. 43: 1164-
particles and antimicrobial activity. J. Am. 1170.

S
Chem. Soc. 127: 2285–2291. [40] Ganesh K.,Dinesh V., Gokavarapu S.,
[33] Son W.K., Youk J.H., Lee T.S., Park W.H., Rajeswari A., Stalin D.T., Karthick Z.,

f
(2004), Preparation of antimicrobial Kapadia V., Shrestha T., Barathy I.A.;
ultrafine cellulose acetate fibers with silver Roy A., Sinha S., (2011), Facile green

[34]
nanoparticles. Macromol. Rapid Commun.
25: 1632–1637.
Sastry M., Mayyaa K.S., Bandyopadhyay
K., (1997), pH dependent changes in the

e o synthesis of gold nanoparticles using

leaf
Cassia
extract of antidiabetic potent
auriculata. Coll. Surf. B.
Biointerf. 87: 159– 163.
optical properties of carboxylic acid

i v
derivatized silver colloid particles. Colloids
Surf. A. 127: 221–228.
[35]

c h
Wang X., Bunkers G.J., (2000), Potent
heterologous antifungal proteins from
cheeseweed (Malva parviflora). Biochem.

r
Biophys. Res. Commun. 279: 669–673.

A
[36] Badri Narayanan K., Sakthivel N., (2008),
Coriander leaf mediated biosynthesis of
gold nanoparticles. Mater. Lett. 62: 4588–
4590.

Cite this article as: B. Sadeghi: Green synthesis of silver nanoparticles using seed aqueous extract of Olea europaea.
Int. J. Nano Dimens. 5(6): 575-581 (Special Issue) 2014.

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