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Role of capping agents in controlling silver

nanoparticles size, antibacterial activity and
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Cite this: RSC Adv., 2016, 6, 36171

potential application as optical hydrogen peroxide
sensor†
B. Ajitha,‡*a Y. Ashok Kumar Reddy,‡b P. Sreedhara Reddy,c Hwan-Jin Jeon*a
and Chi Won Ahn*a

Received 10th February 2016
Accepted 5th April 2016
DOI: 10.1039/c6ra03766f
www.rsc.org/advances
The influence of capping agents on silver nanoparticles (AgNPs) was investigated through a rapid and
single-pot chemical reduction method. Four capping agents were tested: polyethylene glycol (PEG),
ethylenediaminetetraacetic acid (EDTA), polyvinyl pyrrolidone (PVP) and polyvinyl alcohol (PVA). FTIR
studies demonstrated that the formed AgNPs were properly encapsulated by their respective capping
agents. Structural and morphological studies confirmed the following relative average particle sizes:
PEG-AgNPs > EDTA-AgNPs > PVP-AgNPs > PVA-AgNPs. Optical absorption and photoluminescence
studies showed, respectively, a greater absorption blue shift and greater emission intensity for the
smaller capped particles. Zeta potential analysis of the PVA-AgNPs showed a value of
46.6 mV,
indicating their high stability. The PVA-AgNPs were thus not only observed to be the smallest, most
blue-shifted and most stable of the tested AgNPs, but also they displayed the highest antibacterial
activity. The PVA-AgNPs were therefore applied as a localized surface plasmon resonance (LSPR)-based
H2O2 sensor, which is important because the detection of reactive oxygen species such as H2O2 is of
significance in the medical and environmental fields. The sensor based on the PVA-AgNPs successfully
detected H2O2 at concentrations as low as 10
7 M. New biosensors using these NPs should thus find
promising opportunities in a variety of fields.

because of their large operative scattering cross section and

1. Introduction
Recent advancements in nanotechnology have garnered
increasing attention due to the improvements in producing and
utilizing particles whose sizes are in the nanometer regime.
There is particular interest in noble metal nanoparticles due to
their distinctive properties, such as their optical, mechanical,
electronic, chemical and magnetic properties which are entirely
different from corresponding bulk material properties.1–3 Silver
nanoparticles (AgNPs) have been of particular interest owing to
their unique electrical, optical and physicochemical properties
as well as their biomedical applications.4 AgNPs show good
conductivity, chemical stability, catalytic, antibacterial activity
and surface enhanced Raman spectroscopy (SERS) effects, and
also constitute an ideal candidate for molecular labeling
a
Nano-Materials Laboratory, National NanoFab Center, Daejeon, 305-338, Republic of
Korea. E-mail: ajithabondu@gmail.com; hjjeon@nnfc.re.kr; cwahn@nnfc.re.kr
b
Department of Electrical Engineering, Korea Advanced Institute of Science and
Technology, 335 Gwahangno, Yuseong-gu, Daejeon 305-701, Republic of Korea
c
Department of Physics, Sri Venkateswara University, Tirupati-517502, India
† Electronic supplementary information (ESI) available. See
10.1039/c6ra03766f
‡ Authors contributed equally.
surface plasmon resonance (SPR).5,6 Metal nanoparticles in
general can be synthesized by various approaches such as
chemical,7 electrochemical,8 chemical vapor deposition,9
photochemical,10 molecular beam epitaxy,11 and chemical
reduction with and without stabilizing polymers,7,12 which is
a particularly widely used chemical approach. Note that exper-
imental factors, the interaction of metal precursor ions with
reducing agents, and capping agent interface with metal
nanoparticles greatly inuence the size, shape, stability and
physicochemical properties of the metal nanoparticles.13,14
In contrast to bulk material, the corresponding nano-
particles exhibit high surface energy values, due to their small
sizes, and hence oen congregate.15 Therefore, selecting an
appropriate capping agent is oen a prerequisite for stabilizing
nanoparticles.16 Capping agents appear to work by using
a variety of mechanisms, including electrostatic stabilization,
steric stabilization, stabilization by hydration forces, depletion
stabilization and stabilization using van der Waals forces,16 and
a combination of some of these mechanisms may be at work for
certain capping agents, such as branched polyethylenimine
DOI: (BPEI).17 The choice of capping agent for nanoparticles is
crucial, because the capping agent oen inuences various

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properties of the nanoparticle, including its size, shape and
interactions with surrounding solvent.18 Apart from their main
role in stabilizing nanoparticles, some capping agents (e.g.,
citrate) are capable of carrying out the additional role of
reducing metal ions (e.g., silver ions) into metal nanoparticles.19
Therefore, the selection of the capping agent plays a vital role in
the nanoparticle synthesis process, and capping agents also
affect nanoparticle properties.20
Well-stabilized nanoparticles are important for their func-
tions and applications, including their ability to serve as
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sensors of chemicals in the environment. One such chemical of
concern is hydrogen peroxide (H2O2). H2O2 is a strong reducing
agent and widely utilized in the food and pharmaceutical
industries as well as for the bleaching of wood and pulp.
However, H2O2 even in small amounts is highly toxic and is
a serious threat to biological systems.21 Therefore it has been an
upsurge of interest to develop H2O2 sensors in order to detect
H2O2 promptly and accurately. The development of
nanomaterial-based sensing devices and detection methods has
largely relied on the use of localized surface plasmon resonance
(LSPR) since the resonant wavelength is very sensitive to the
environment surrounding the nanoparticles being analyzed.
Noble metal nanostructures such as gold and silver are appli-
cable as LSPR-based optical and chemical sensors/biosensors
and are expected to be of use in high-sensitivity detection of
target molecules in medical and food control applications.22,23
Due to the simplicity and low cost of fabricating hydrogen
peroxide sensors, the analytical detection of hydrogen peroxide
is extensively used in pharmaceutical, environmental, indus-
trial and clinical research.24 Recently, Endo et al.25 developed an
LSPR-based optical biosensor using PVP-capped AgNPs for the
detection of hydrogen peroxide. Tagad et al.26 constructed
a H2O2 sensor based on silver nanoparticles stabilized by
polysaccharides.
In the present study, we synthesized spherical AgNPs in the
presence of different stabilizing capping agents through
a simple chemical reduction method, and we optimized various
features of the AgNPs produced. The capping agents tested
included polyethylene glycol (PEG), ethylenediaminetetraacetic
acid (EDTA), polyvinylpyrrolidone (PVP) and polyvinyl alcohol
(PVA), and we determined the effects of these capping agents on
the sizes of the resulting AgNPs. In addition, as part of the effort
to develop improved detectors of H2O2, the current work
focused on applying our synthesized AgNPs as an LSPR-based
H2O2 optical sensor, and the characteristics of this sensor was
evaluated by carrying out a UV-visible spectrophotometric
analysis. We also evaluated the synthesized AgNPs for their
antibacterial activity.
2. Experimental
2.1 Materials
Silver nitrate (AgNO3), sodium borohydride (NaBH4), ethanol
(C2H5OH), PEG (C2nH4n+2On+1), PVP ((C6H9NO)n), EDTA
(C10H16N2O8), PVA ((C2H4O)n) and sodium hydroxide (NaOH)
were purchased from Sigma Aldrich. All the chemicals were of
analytical reagent grade and used without further purication.
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2.2 Procedure used to synthesize silver nanoparticles with
different capping agents
In a typical procedure, say when using the capping agent PEG,
the total volume of the reaction solution is xed at 150 ml. 1 ml
of 1% PEG aqueous solution is slowly added drop wise for 5 min
into 98.5 ml of ultrapure water in a 250 ml three-necked ask,
while heating at 75  C in an oil bath under stirring. Then, 50 ml
of 0.01 M aqueous AgNO3 solution is injected in to the above
mixture using a programmable syringe pump at a rate of 50 ml
h
1 and 150 ml of sodium hydroxide (0.02 M) solution is added
rapidly at once. This is followed by the injection of ice cold
freshly prepared aqueous NaBH4 solution (350 ml, 0.05 M) using
a syringe pump with infusion rate set at 50 ml min
1 upon
vigorous stirring; the color of this suspension changes to deep
yellow. The reaction was allowed to proceed for two hours, aer
which the solution was cooled to room temperature. The
products were obtained by repeated centrifugation for thrice
and washed with water and ethanol twice at 4  C in order to
remove unreacted constituents and were nally subjected to
vacuum drying. The same procedure was used with the other
capping agents, which included EDTA, PVP and PVA. The nal
products (4.5 mg of each sample) were transferred into amber
bottles and stored for further analysis.
2.3 Characterization of the materials
The synthesized AgNPs were characterized comprehensively by
using a variety of techniques. The crystalline structure of the
synthesized AgNPs was analyzed using a Seifert 3003 TT X-ray
diffractometer with Cu Ka radiation (k ¼ 0.1546 nm). The
surface morphology of the AgNPs was visualized using a ZEISS,
SUPRA 55 eld emission scanning electron microscope
(FESEM). A transmission electron microscopy (TEM) study was
conducted using a Technai TF30 ST eld emission transmission
electron microscope (FE-TEM) working at an accelerating
voltage of 300 kV. UV–visible absorption spectra were acquired
on a Perkin Elmer Lambda 950 UV-Vis-NIR spectrophotometer
with the wavelength set in the range 200–800 nm at a resolution
of 0.2 nm. Photoluminescence spectra were recorded using
a Horiba Jobin–Yvon Fluorolog-3 spectrouorometer (FL3-
22PTI). Fourier transform infrared spectroscopy (FTIR) spectra
of AgNPs were taken with an ATR-FTIR Bruker Vertex-80 spec-
trometer. The zeta potential of the prepared AgNPs was
measured using a Nanopartica instrument (HORIBA).
2.4 Antibacterial assay of the synthesized AgNPs
The antibacterial activities of the synthesized AgNPs with
different capping agents were tested against the bacterial
isolates such as Escherichia coli and Pseudomonas spp. The
antibacterial properties of these different AgNPs were investi-
gated by employing standard Kirby–Bauer's disc diffusion
method.27 The above-mentioned bacterial pathogens were
cultured overnight in nutrient broth at 37  C. The nutrient agar
plates were prepared by dissolving the nutrient agar in distilled
water and autoclaved and then allowed for solidication before
adding the cultures through lawn culture method. Then,
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Whatman no. 1 lter paper discs impregnated with a volume of
6 ml of the AgNPs synthesized with different capping agents and
dried under aseptic conditions, were placed in separate appro-
priately labeled plates. The plates were incubated at 37  C for 24
h to assess the activity of the AgNPs. The size (in mm) of the
clear zone of inhibition around each disc aer the incubation
period was recorded.
2.5 LSPR-based hydrogen peroxide detection method using
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PVA-capped AgNPs
The optical characteristics of PVA-capped AgNPs serving as an
LSPR-based sensor of hydrogen peroxide were evaluated using
various concentrations of H2O2 as specied in the previously
used protocol.25 At rst, a volume of 1 ml of a hydrogen peroxide
solution of a specied concentration was diluted with 20 mM
phosphate buffer (pH 7.4), and the resulting solution was added
to PVA-capped AgNPs in a quartz cuvette and thoroughly mixed
for evaluation of optical characteristics using a UV-vis spectro-
photometer. The changes in the optical spectra of the AgNPs at
a wavelength of about 425 nm were monitored at regular
intervals of time (0, 5, 10, 15, 20 and 30 min).
3. Results and discussion
3.1 Structural analysis
The structures of the AgNPs synthesized with different capping
agents were investigated using X-ray diffraction (XRD) analysis.
The XRD spectra with 2q values ranging from 20 to 80 depic-
ted four peaks corresponding to diffraction from the (111),
(200), (220) and (311) planes (Fig. 1). Diffraction from the (111)
plane was strongest. All the peaks were indexed based on the
face-centered cubic (FCC) lattice of silver, and were consistent
with the standard JCPDS (no. 04-0783) data. The average crys-
tallite size was estimated from the FWHM of the diffraction
peak using the Debye–Scherrer formula28
Fig. 1 Representative XRD profiles of synthesized silver nanoparticles
with different capping agents.
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D ¼ Kl/b cos q, (1)
where K denotes Scherrer's constant (K ¼ 0.94), D corresponds
to crystallite size, b is the full width at half maximum (FWHM)
of the XRD peak and is expressed in radians, and q is the
position of the diffraction peak. The FWHM was calculated
using (111) preferred orientation and the average crystallite size
was found to range from a high of 39 nm for the PEG-capped
AgNPs to a low of 26 nm for the PVA-capped AgNPs.
3.2 Morphological studies
FESEM measurements were taken to obtain the particle size
distribution of the as-synthesized AgNPs. The FESEM images
(Fig. 2) clearly showed the formation of spherical AgNPs
synthesized with the different capping agents.
The PEG-capped AgNPs were observed to agglomerate to
a modest extent into larger particles, while the others agglom-
erated to lesser extents. The least agglomeration was observed
for the PVA-capped AgNPs, which for the most part formed
isotropic spherical nanoparticles. The average particle sizes of
the PEG, EDTA, PVP and PVA capped AgNPs were found to be
44, 39, 35 and 31 nm, respectively. Therefore, PVA capped
AgNPs are optimized as a smallest particle size.
The shapes and sizes of the synthesized AgNPs with different
capping agents were further examined by using TEM. The
typical TEM micrographs of PEG and PVA-capped AgNPs are
shown in Fig. 3(a) and (b), and the average particle size was
found to be 41 nm for the PEG-capped AgNPs and 27 nm for the
PVA-capped AgNPs.
In these micrographs, the more spherical isotropic particles
appeared smaller in size whereas the nonspherical ones
appeared larger. For the PEG-capped case, the smaller particles
apparently agglomerated to form larger nonspherical particles,
perhaps because of a decreased interaction between the PEG and
AgNPs relative to the interactions between the other capping
agents and the AgNPs. Less agglomeration was observed for the
Fig. 2FESEM images of AgNPs prepared with (a) PEG, (b) EDTA, (c)
PVP and (d) PVA.
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Fig. 3 TEM images of AgNPs synthesized with (a) PEG and (b) PVA. (c)
Typical HR-TEM and (d) SAED pattern of AgNPs prepared with PVA.
PVA-capped AgNPs than for the others, and the PVA-capped
AgNPs were observed to be generally spherical and homoge-
neously dispersed. Multiple and equispaced lattice fringes were
clearly observed in the high-resolution TEM of the PVA-capped
AgNPs (Fig. 3(c)). The lattice spacing (d) of the (111) plane was
found to be 0.23 nm and is consistent with JCPDS data (no. 04-
0783). Finally, the PVA-capped AgNPs were found to be smaller
than the AgNPs made with the other capping agents. The PVA-
capped AgNPs were also analyzed by electron diffraction
directly on the transmission electron microscope. The selected-
area electron diffraction (SAED) pattern (Fig. 3(d)) showed
obvious bright circular rings mixed with several diffraction spots
attributed to the polycrystalline structure of the formed AgNPs.
3.3 Optical properties
UV-vis absorption spectroscopy is a fundamental technique for
assessing the formation of stable AgNPs, since metal nano-
particles have been shown to exhibit an intense absorption peak
due to the surface plasmon resonance (SPR) phenomenon
(collective oscillation of surface electrons) at 395–425 nm.29
According to Mie's theory30 only a single SPR band is expected in
the absorption spectrum of spherical nanoparticles, whereas
two or more SPR bands are expected for anisotropic nano-
particles, depending on the specic shapes of the particles. The
number of SPR peaks increases as the symmetry of the nano-
particles decreases.31
Fig. 4(a) displays the absorption spectra of the AgNPs
prepared with different capping agents. A single and fairly
symmetrical SPR band was exhibited by each of the AgNP
samples in the range of 440–420 nm. AgNPs synthesized with
PEG, EDTA, PVP and PVA yielded maximum absorption peaks at
445, 438, 431 and 425 nm, respectively, indicating the formation
of relatively small nanoparticles with narrow size distributions.
The SPR data was well correlated with the shapes of the
synthesized nanoparticles in the micrographs described above.
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From this, we can state that the relatively short wavelength at
maximum absorption for the PVA-capped AgNPs was due to the
relatively small average size of the particles.
To further explore the size-dependent optical properties, we
carried out photoluminescence studies. Fig. 4(b) shows the PL
spectra of the AgNPs synthesized with the PEG, EDTA, PVP and
PVA capping agents, respectively. It has been suggested that the
photoemission wavelength is independent of the particle size,
while the intensity increases with decreasing particle size.32
Irrespective of the capping agent used, all AgNPs excited with
400 nm wavelength light yielded a maximum emission at
a similar wavelength, mainly located in the visible region at 450
nm. The PVA-capped AgNPs yielded the most intense emission,
consistent with their smallest average particle size. As the
AgNPs were excited in the SPR range, the radiation absorbed by
these nanoparticles is thought to be radiative energy transfer to
the emitting particles.
3.4 FTIR studies
In order to determine the possible functional groups involved in
reduction and capping process and their unique interactions
with AgNPs, FTIR spectroscopy studies were undertaken.
Fig. 5(a) shows the FTIR spectra of the AgNPs synthesized with
the PEG, EDTA, PVP and PVA capping agents, respectively. PEG-
capped AgNPs yielded a broad peak centered around 3412 cm
1
attributed to the stretching vibration of –OH group. The spectral
bands at 1070 cm
1, 1260 cm
1, 1380 cm
1, and 1587 cm
1
arose from the C–OH stretching, C–O–C asymmetric stretching,
alkyl C–H deforming vibrations, and C–H mode vibrations,
respectively. The small peaks at 530 cm
1 and 415 cm
1
correspond to the AgNPs bonding with oxygen of hydroxyl
groups of PEG units, and C–C skeletal vibrations were observed
at 989 and 840 cm
1. All of the above spectral peaks are in
accordance with the previous report and reveal that PEG stabi-
lized the NPs through steric and electrostatic interactions of
–OH groups.33 EDTA-capped AgNPs yielded absorptions at 1061
cm
1, 1401 cm
1, 1632 cm
1 and 3446 cm
1, which were
assigned to C–O single-bond vibrations, asymmetric carboxylate
group stretching, amide-I C]O stretching, and O–H stretching
vibrations, respectively.34 This IR spectrum strongly conrmed
the capping of the AgNPs through direct bonding of EDTA.
Each repeating unit of PVP has two functional groups, a C–N
group and a C]O group. These groups, due to their N and O
atoms, have great affinity for silver ions and can coordinate
metallic silver.35 PVP-coated AgNPs yielded main absorption of
C–N peak of pure PVP at 1019 cm
1 which is separated into two
peaks at 1056, and 1007 cm
1.36 Other peaks at 1378 cm
1, 1631
cm
1, 3491 cm
1 are characteristic of bond vibrations of the
NO3
1 group, C]O bonds, and O–H stretching vibrations,
respectively.36 Thus, the FTIR spectra showed the presence of
molecular interactions between AgNPs and PVP chain.
PVA-capped AgNPs yielded strong absorption peaks at 3446
cm
1, 1641 cm
1, 1420 cm
1, and 1062 cm
1, which were
ascribed to typical hydroxyl bands, symmetric stretching of
carboxylate anion, O–H and C–H bending, and C–O stretching,
respectively.7 In addition, small bands were also observed at
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Fig. 4 (a) UV-vis absorption spectra and (b) PL spectra of the synthesized AgNPs with the various capping agents.
2948 and 2897 cm
1, corresponding to the asymmetric and the
symmetric stretching of –CH2–, respectively. As a result, we may
conclude that all of the synthesized AgNPs were properly
encapsulated by their capping agents.
3.5 Zeta potential analysis
Zeta potential is an important parameter for the assessing the
stability of AgNPs in aqueous suspensions. Particles that have
a large negative or positive zeta potential are likely to repel each
other, and thereby have enhanced suspension stability.
However, if the particles have low zeta potential values, then
attractive forces between the particles become dominant and
particles undergo aggregation and occulation. In general,
a minimum value of 30 mV for the zeta potential is necessary
to validate a stable nanosuspension.37 Fig. 5(b) shows the zeta
potential values of the synthesized AgNPs with different
capping agents and indicates the high stability of the prepared
AgNPs. Moreover, the zeta potential results revealed the
synthesized PVA-AgNPs to be more stable than the other AgNP
Fig. 5 (a) FTIR spectra and (b) zeta potential values for AgNPs synthesized with different capping agents.
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samples. Additionally, the negative zeta potential value (
46.6
mV) of PVA-AgNPs also suggests an active role of the capping
agent in augmenting the stability of the AgNPs for a longer
period of time than other capped AgNPs.
3.6 Formation of AgNPs with capping agents
Different capping agents (polyethylene glycol (PEG), ethyl-
enediaminetetraacetic acid (EDTA), polyvinylpyrrolidone (PVP)
and polyvinyl alcohol (PVA)) were used for the synthesis of
AgNPs and formed AgNPs were encapsulated by their own
capping agent. In case of PEG, the hydroxyl group acts as
a capping agent and can make a cover on the surface of AgNPs.
This is probably due to the presence of van der Waals forces
between negatively charged oxygen groups present in PEG and
positively charged that surround the surface of the inert
AgNPs.38 The possible interaction between the positively
charged Ag and PEG is shown in Fig. S1(a).† The EDTA molec-
ular structure allows it to completely surround a metal atom
(chelating effect) and avoids the metal from reacting with other
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molecules by forming cage-like structure as EDTA is a bifunc-
tional chelating agent.39 Since, chelating agent may bond to
a metal ion in more than one place simultaneously and increase
its stability. Herein, the EDTA bond up with the silver ion and
the schematic representation of silver ion wrapped by EDTA is
shown in Fig. S1(b).†
PVP has a polyvinyl skeleton with polar groups, act as
a capping agent for dissolved metallic salts through steric and
electrostatic stabilization of amide groups of the pyrrolidone
rings. PVP donates loan pair electrons of oxygen and nitrogen
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atoms to silver ions and thus forms the Ag ions and PVP
complex structure.40 The schematic reaction between silver
cation and PVP is depicted in Fig. S1(c).† PVA is a non-toxic
green polymer and it reacts with silver nitrate resulting in the
formation of Ag+ ions bonded to PVA polymer structure. As PVA
is heated while stirring, the reactive PVA molecules with acti-
vated OH-groups free from H-bonding are formed.41 Thus, these
active OH-groups act as head-groups to adsorb Ag+ cations and
drive a directional growth, Ag+ / Ag0, which eventually produce
a Ag-metal attached to the PVA surface (Fig. S1(d)†).
3.7 Antibacterial studies
The antibacterial properties of the AgNPs with different capping
agents (PEG, EDTA, PVP and PVA) were analyzed by means of
Kirby–Bauer's disc diffusion method on the basis of the zone of
inhibition. The antibacterial activity of the AgNPs is solely due
to physical effects, such as adsorption of AgNPs to the cell wall,
interaction of Ag ions with the cell wall protein, denaturation of
the cell wall proteins, and the formation of porous structures
along with severe changes in the membrane structure of the
bacteria, which all increase the permeability and accumulation
of external uids. In particular, AgNPs can damage the structure
of the cell membrane by anchoring onto and penetrating the
membrane through electrostatic attractions, and hence deacti-
vate certain membranous enzymes, dissipate the proton motive
force, and nally cause cell lysis.42,43 Fig. 6 shows the
Fig. 6 Bactericidal activity of AgNPs with different capping agents against E. coli (a–d) and Pseudomonas spp. (e–h).
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Table 1 Antibacterial activity of AgNPs, measured against different
pathogenic organisms
Zones of inhibition (mm)
AgNPs capping agent Escherichia coli Pseudomonas spp.
PEG 3 3
EDTA 4 5
PVP 6 6
PVA 7 8
antibacterial activity of the AgNPs synthesized with the various
capping agents. Martı́nez-Castañón et al.44 reported that rela-
tively small particles display relatively high antibacterial
activity. The PVA-capped AgNPs, shown above to be small,
indeed showed better antibacterial efficacy than the other
AgNPs (Fig. 6). Also, the high aspect ratio and the crystallo-
graphic structure of the nanoparticles govern their antimicro-
bial potency, which are favored by {111} facets.45
The AgNPs exhibited better antibacterial efficacy by inhibit-
ing the growth and division of pathogenic bacteria like Escher-
ichia coli and Pseudomonas spp. In general, AgNPs destabilize
the outer membrane and split the plasma membrane, thereby
causing reduction of intracellular ATP.46 The sizes in millime-
ters of the zones of inhibition of the respective discs loaded with
different concentrations of AgNPs against the tested pathogenic
bacteria are listed in Table 1. Of the different capping agents
tested, the PVA-capped AgNPs yielded the largest such zones of
inhibition, which revealed these AgNPs to be novel antibacterial
agents due to their small average size.
3.8 Evaluation of PVA-capped AgNPs as LSPR-based optical
hydrogen peroxide sensors
The catalytic reaction between PVA-capped AgNPs and
hydrogen peroxide would be expected to inevitably result in the
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Fig. 7 Possible mechanism and schematic representation of (a) the reaction between PVA-coated AgNPs and H2O2, and (b) the decrease of
absorbance intensity with increasing H2O2 concentration. (c) Optical absorption characteristics of different concentrations of PVA-capped
AgNPs. (d) LSPR optical absorbance spectra of AgNPs as a function of time after the addition of 10
3 M H2O2. (Inset: AgNPs solution (i) before and
(ii) after addition of H2O2). (e) The correlation between the optical absorbance strength changes after 30 min with respect to different
concentrations of H2O2 added to the PVA-AgNPs. (f) Relationship between the absorbance intensity and reaction time for different concen-
trations of H2O2.

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decomposition of hydrogen peroxide, and induce the degrada-
tion of clustered AgNPs through PVA layer with their oxidation
(Fig. 7(a) and (b)). The dependence of the absorbance of the
PVA-capped AgNPs on its concentration is shown in Fig. 7(c).
The absorption intensity of the PVA-capped AgNP increased as
its concentration was increased from 0.0005% to 0.001% and to
0.002%. As a result, the 0.002% concentration of the PVA-
capped AgNP solution was chosen as a hydrogen peroxide
sensor.
Fig. 7(d) shows the change in the LSPR-based optical H2O2
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sensor absorbance characteristics of the PVA-capped AgNPs
with time due to the introduction of 1 mM (10
3 M) H2O2. As
shown in the inset of Fig. 7(d), the signature yellow color (vial
(i)) disappeared and nally became colorless (vial (ii)) aer 30
min. The absorbance was observed to decrease with time of
reaction, and aer 30 min a drastic change in its optical
absorbance characteristics was observed. The observed
decrease in the absorbance intensity was attributed to the
decrease in the AgNP concentration. The change in color of the
AgNP solution and decrease of the LSPR peak absorbance were
attributed to the ability of silver to catalyze the decomposition
of hydrogen peroxide.47 Hence, a notable change in the intensity
of the LSPR absorbance was possibly observed.
In order to prove that the catalytic reaction involving hydrogen
peroxide was the major reason for the decrease in the intensity of
the LSPR absorbance with time, a control in which only phos-
phate buffer and distilled water were added to the PVA-capped
AgNPs was also analyzed. In this case, no change in the inten-
sity of the LSPR absorbance was observed. Based on the afore-
mentioned mechanism of the LSPR-based optical sensors and
the experimental conditions, an assessment of the calibration
characteristics was performed. For this purpose, different
concentrations of hydrogen peroxide were added to the PVA-
capped AgNP solution and the change of the absorbance with
time was monitored. The change in the intensity of the absor-
bance aer 30 minutes of reaction time with respect to different
concentrations of H2O2 (Fig. 7(e)), and change in absorbance
strength with reaction time for different H2O2 concentrations
were measured (Fig. 7(f)). Furthermore, the detection limit of this
optical sensor was determined to be 10
7 M.
4. Conclusions
In the present investigation, AgNPs were synthesized using
different capping agents by using a simple chemical reduction
route. The X-ray diffraction analysis revealed a face-centered
cubic lattice for all four tested types of capped AgNPs, but
indicated that the average crystallite size was greatest for the
PEG-capped nanoparticles and smallest for the PVA-capped
ones. Morphological studies revealed the formation of spheri-
cally shaped particles, and visually conrmed the PVA-capped
particles to have the smallest diameters. Optical studies
revealed the blue shi trend due to the decrease of particle size
with different capping agents. The PVA-capped AgNPs were not
only observed to be the smallest and most blue-shied of the
tested AgNPs, but were also observed to be the most stable and
to display the highest antibacterial activity. For these reasons,
36178 | RSC Adv., 2016, 6, 36171–36179
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Paper
PVA-capped AgNPs were chosen for use in an LSPR-based
hydrogen peroxide sensor. The LSPR absorbance strength was
observed to change markedly with changes in the hydrogen
peroxide concentration and time of reaction due to the catalytic
degradation of the AgNPs. The ability of AgNPs to catalyze the
reduction of H2O2 was thus successfully applied to the fabri-
cation of an optical sensor of H2O2. Furthermore, this detection
method is simple, relatively inexpensive, and reliable, and can
be suitably employed in medical and environmental
monitoring.
Acknowledgements
This research was supported by the National Research Foundation
of Korea (NRF-2015K1A4A3047100, NRF-2015M3A7B6027973, and
NRF-2015M3A6B3068660).
References
1 J. Prikulis, F. Svedberg and M. Krall, J. Nanosci. Lett., 2004, 4,
115–118.
2 S. He, J. Yao, P. Jiang, D. Shi, H. Zhang, S. Pang and H. Gao,
Langmuir, 2001, 171, 571–575.
3 B. Wiley, Y. Sun, B. Mayers and Y. Xia, Chem.–Eur. J., 2005,
11, 454–463.
4 A. Ravindran, P. Chandran and S. S. Khan, Colloids Surf., B,
2013, 105, 342–352.
5 C. N. Lok, C. M. Ho, R. Chen, Q. Y. He, W. Y. Yu, H. Sun,
P. K. H. Tam, J. F. Chiu and C. M. Che, J. Proteome Res.,
2006, 5, 916–924.
6 C. F. Mao and M. A. Vannice, J. Catal., 1995, 154, 230–244.
7 B. Ajitha, Y. A. K. Reddy and P. S. Reddy, Powder Technol.,
2015, 269, 110–117.
8 S. A. Vorobyova, A. I. Lesnikovich and N. S. Sobal, Colloids
Surf., A, 1999, 152, 375–379.
9 N. Satoh and K. Kimura, Bull. Chem. Soc. Jpn., 1989, 62, 1758–
1763.
10 Z. Li, Y. Li, X. F. Qian, J. Yin and Z. K. Zhu, Appl. Surf. Sci.,
2005, 250, 109–116.
11 D. W. Bahnemann, Isr. J. Chem., 1993, 33, 115–136.
12 L. M. L. Marzan and A. P. Philipse, J. Phys. Chem., 1995, 99,
15120–15128.
13 W. Chen, W. Cai, L. Zhang and G. Wang, J. Colloid Interface
Sci., 2001, 238, 291–295.
14 S. Sengupta, D. Eavarone, I. Capila, G. L. Zhao, N. Watson
and T. Kiziltepe, Nature, 2005, 436, 568–572.
15 C. N. R. Rao, G. U. Kulkarni, P. J. Thomas and P. P. Edwards,
Chem. Soc. Rev., 2000, 29, 27–35.
16 K. D. Kim, D. N. Han and H. T. Kim, Chem. Eng. J., 2004, 104,
55–61.
17 A. M. E. Badawy, K. G. Scheckel, M. Suidan and T. Tolaymat,
Sci. Total Environ., 2012, 429, 325–331.
18 P. Raveendran, J. Fu and S. L. Wallen, J. Am. Chem. Soc., 2003,
125, 13940–13941.
19 A. Pyatenko, M. Yamaguchi and M. Suzuki, J. Phys. Chem. B,
2005, 109, 21608–21611.
This journal is © The Royal Society of Chemistry 2016

Paper
20 J. N. Kuhn, C. K. Tsung, W. Huang and G. A. Somorjai, J.
Catal., 2009, 265, 209–215.
21 K. R. Hoyt, A. J. Gallagher, T. G. Hastings and I. J. Reynolds,
Neurochem. Res., 1997, 22, 333–340.
22 T. Endo, R. Ikeda, Y. Yanagida and T. Hatsuzawa, Anal. Chim.
Acta, 2008, 611, 205–211.
23 N. Nath and A. Chilkoti, J. Fluoresc., 2004, 14, 377–389.
24 F. Wang, R. Yuan, Y. Chai and D. Tang, Anal. Bioanal. Chem.,
2007, 387, 709–717.
25 T. Endo, Y. Yanagida and T. Hatsuzawa, Measurement, 2008,
Published on 06 April 2016. Downloaded by Freie Universitaet Berlin on 26/03/2018 20:47:32.
41, 1045–1053.
26 C. K. Tagad, H. U. Kim, R. C. Aiyer, P. More, T. Kim,
S. H. Moh, A. Kulkarni and S. G. Sabharwal, RSC Adv.,
2013, 3, 22940–22943.
27 A. L. Barry, F. Garcia and L. D. Thrupp, Am. J. Clin. Pathol.,
1970, 53, 149–158.
28 B. D. Cullity, Elements of X-ray Diffraction, Addison-Wesley
Pub Inc, MA, 2nd edn, 1978.
29 P. Mulvaney, Langmuir, 1996, 12, 788–800.
30 G. Mie, Ann. Phys., 1908, 25, 377–445.
31 I. O. Sosa, C. Noguez and R. G. Barrera, J. Phys. Chem. B,
2003, 107, 6269–6275.
32 S. Ashokkumar, S. Ravi, V. Kathiravan and S. Velmurugan,
Spectrochim. Acta, Part A, 2014, 121, 88–93.
33 S. Tunc and O. Duman, Colloids Surf., A, 2008, 37, 93–99.
34 S. Mondal and S. Verma, Z. Anorg. Allg. Chem., 2014, 640,
1095–1101.
35 P. Silvert, R. Herrera-urbina, N. Duvauchelle,
V. Vijayakrishnan and K. T. Elhsissen, J. Mater. Chem.,
1996, 6, 573–577.
This journal is © The Royal Society of Chemistry 2016
View Article Online
RSC Advances
36 Z. Zhang, B. Zhao and L. Hu, J. Solid State Chem., 1996, 121,
105–110.
37 C. Jacobs and R. H. Muller, Pharm. Res., 2002, 19, 189–194.
38 P. Dallas, V. K. Sharma and R. Zboril, Adv. Colloid Interface
Sci., 2011, 166, 119–135.
39 K. P. McGrath, B. T. Do and J. G. MacDonald, US Pat.,
US2005/0084464 A1, 2005.
40 Z. Jovanovic, A. N. Radosavljevic, N. Bibic, M. Siljegovic,
Z. M. Kacarevic-Popovic and V. B. Miskovic-Stankovic,
Radiat. Phys. Chem., 2012, 81, 1720–1728.
41 A. Gautam, P. Tripathy and S. Ram, J. Mater. Sci., 2006, 41,
3007–3016.
42 J. R. Morones, J. L. Elechiguerra, A. Camacho, K. Holt,
J. B. Kouri, J. T. Ramirez and M. J. Yacaman,
Nanotechnology, 2005, 16, 2346–2353.
43 J. Kim, E. Kuk and K. Yu, Nanomedicine, 2007, 3, 95–101.
44 G. A. Martı́nez-Castañón, N. Niño-Martı́nez, F. Martı́nez-
Gutierrez, J. R. Martı́nez-Mendoza and F. Ruiz, J. Nanopart.
Res., 2008, 10, 1343–1348.
45 S. Shrivastava, T. Bera, A. Roy, G. Singh, P. Ramchandrarao
and D. Dash, Nanotechnology, 2007, 18, 225103–225112.
46 M. J. Hajipour, K. M. Fromm, A. A. Ashkarran,
D. J. Aberasturi, I. R. Larramendi, T. Rojo, V. Serpooshan,
W. J. Parak and M. Mahmoudi, Trends Biotechnol., 2012,
30, 499–511.
47 T. Pal, D. S. Maity and A. Ganguly, Talanta, 1988, 35, 658–
660.
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