Journal of Colloid and Interface Science 336 (2009) 117–124

Contents lists available at ScienceDirect

Journal of Colloid and Interface Science

www.elsevier.com/locate/jcis

Lasting antibacterial activities of Ag–TiO2/Ag/a-TiO2 nanocomposite thin film

photocatalysts under solar light irradiation
O. Akhavan *
Department of Physics, Sharif University of Technology, P.O. Box 11155-9161, Tehran, Iran

a r t i c l e i n f o a b s t r a c t

Article history: Photodegradation of Escherichia coli bacteria in presence of Ag–TiO2/Ag/a-TiO2 nanocomposite film with
Received 15 December 2008 an effective storage of silver nanoparticles was investigated in the visible and the solar light irradiations.
Accepted 20 March 2009 The nanocomposite film was synthesized by sol–gel deposition of 30 nm Ag–TiO2 layer on 200 nm ana-
Available online 31 March 2009
tase(a-)TiO2 film previously doped by silver nanoparticles. Both Ag/a-TiO2 and Ag–TiO2/Ag/a-TiO2 films
were transparent with a SPR absorption band at 412 nm. Depth profile X-ray photoelectron spectroscopy
Keywords: showed metallic silver nanoparticles with diameter of 30 nm and fcc crystalline structure were self-accu-
Silver nanoparticles
mulated on the film surface at depth of 5 nm of the TiO2 layer and also at the interface of the Ag–TiO2 and
Titanium oxide photocatalyst
Solar light
a-TiO2 films (at depth of 30 nm). Both OH bounds and H2O contents were concentrated on the film sur-
Photodegradation face and at the interface, as a profit in releasing more ionic (not metallic) silver nanoparticles. Antibac-
Escherichia coli bacteria terial activity of the nanocomposite film against E. coli bacteria was 5.1 times stronger than activity of
the a-TiO2, in dark. Photo-antibacterial activity of the nanocomposite film exposed by the solar light
was measured 1.35 and 6.90 times better than activity of the Ag/a-TiO2 and a-TiO2, respectively. The main
mechanism for silver ion releasing was inter-diffusion of water and silver nanoparticles through pores of
the TiO2 layer. Durability of the nanocomposite film was at least 11 times higher than the Ag/a-TiO2 film.
Therefore, the Ag–TiO2/Ag/a-TiO2 photocatalyst can be nominated as one of the effective and long-lasting
antibacterial nanocomposite materials.
Ó 2009 Elsevier Inc. All rights reserved.

1. Introduction
Nanocomposite heterogeneous photocatalysts working based on
the interaction of light with the dispersed metallic nanoparticles
have attracted great interest due to their high and broad photocata-
lytic activities. Among various metal-oxide materials being devel-
oped for photocatalytic applications, titanium dioxide (TiO2) has
received great attentions because of its chemical stability and high
reactivity under UV light irradiation (k < 390 nm) [1–5]. When ana-
þ
tase TiO2 is exposed to UV light, holes ðhvb Þ and excited electrons
ðe
cb Þ are generated. The holes are trapped by water (H2O) or hydroxyl
groups (OH) adsorbed on the surface to generate hydroxyl radicals
(OH) [6,7] which is a powerful and indiscriminate oxidizing agent
for degrading a wide range of organic pollutants [8–10].
After the first study on bactericidal properties of anatase TiO2
[11], many works on inactivation of bacteria, molds, viruses, and
even cancer cells have been reported [12–17]. Since the post-sep-
aration of the powder catalyst in a slurry system is a main disad-
vantage [4,18], the immobilized TiO2 with high surface area (e.g.,
porous TiO2 thin film) is more favorable in the antimicrobial appli-
cations [17,19–22].
* Fax: +98 21 66022711.
E-mail address: oakhavan@sharif.edu
On the other hand, due to the wide band gap of the TiO2
(3.2 eV), it can only be applied in the UV region of the solar spec-
trum (5% of the solar energy), while the visible light contains
about 45% of the solar energy. Metal doping has long been known
to be one of the most effective ways to change the intrinsic band
structure of TiO2, and consequently, to improve its visible light
sensitivity [23–29] as well as increase its photocatalytic activity
under UV irradiation [30–32]. Among various dopants, noble met-
als (especially Ag) have received much attention for this purpose.
However, before anything, silver itself is known as one of the
most interesting antibacterial materials [17,33–37]. It is generally
believed Ag+ can bind to bacterial cell wall membrane (slightly
negative), damage it and so alter its functionality [38–40]. Ag+
can interact with thiol groups in proteins, resulting in inactivation
of respiratory enzymes and leading to the production of reactive
oxygen species [41]. In addition, because of interaction between
the Ag+ and DNA structure of bacteria their multiplication may
be prevented [38,39]. Ag particles of less than 10 nm are more
toxic to bacteria such as Escherichia coli [42,43]. Therefore,
researchers strongly aim to revive the bactericidal applications of
silver (especially silver nanoparticles), due to evolution of new
resistant bacteria against the common antibiotics [44].
As mentioned, silver particles enable to activate visible light
excitation of TiO2 [45]. Recently, it was shown that Ag doped

0021-9797/$ - see front matter Ó 2009 Elsevier Inc. All rights reserved.
doi:10.1016/j.jcis.2009.03.018
118 O. Akhavan / Journal of Colloid and Interface Science 336 (2009) 117–124
TiO2 highly improved photocatalytic inactivation of bacteria
[46–49]. For instance, only 1 wt% Ag in TiO2 reduced the reaction
time required for complete removal of 107 cfu/ml E. coli from 65
to 16 min in UV-A light [47]. It is generally believed that Ag nano-
particles enhances photoactivity of TiO2 by lowering the recombi-
nation rate of its photo-excited charge carriers and/or providing
more surface area for adsorption [50,51]. Visible light absorption
by surface plasmon resonance of Ag nanoparticles is also thought
to induce electron transfer to TiO2 resulting in charge separation
and thus activation by visible light [45,52]. Therefore, Ag–TiO2
nanocomposites show great promise as efficient and visible light
response photocatalytic materials in future.
In practical applications, both high antibacterial activity and
low silver release are two important characteristics for silver-
based materials. High release level of silver, especially for silver-
based bulk materials, leads to shortening the effective life of anti-
bacterial activity [53]. If Ag nanoparticles and nanostructures with
high antibacterial activities [54–58] are immobilized on porous
matrixes, the release time of silver can be delayed for a long time
[59,60] so that these kinds of silver-supported materials will be
of great potentials for bactericidal application.
Recently, it was shown that the water contents of the sol–gel
silica films result in accumulation of majority of the metallic nano-
particles on surface of the aqueous and porous silica film during a
low-temperature (100 °C) heat treatment process [61]. Therefore,
the same process can be used to synthesize surface accumulated
Ag nanoparticles on aqueous TiO2 host layer to obtain a photocat-
alyst thin film with a good sensitivity to the solar light and with
strong and long life antibacterial activity.
In this work, antibacterial activity and durability of sol–gel syn-
thesized Ag–TiO2 nanocomposite layer deposited on Ag/a-TiO2
(anatase TiO2) thin film was investigated against E. coli bacteria,
in dark, visible light and the solar light. Self-accumulation of Ag
nanoparticles on the film surface was examined by depth profile
X-ray photoelectron spectroscopy (XPS) and scanning electron
microscopy (SEM) analyses. Moreover, the role of Ag nanoparticles
in increasing the photocatalytic efficiency of the Ag–TiO2/Ag/a-
TiO2 nanocomposite thin film was investigated by using XPS and
antibacterial analyses. The behavior and durability of silver ion re-
lease from the thin films were studied in an acidic solution. A
mechanism was also proposed for the ion release process.
2. Materials and methods
2.1. Preparation of anatase TiO2 thin films
Sol–gel technique was utilized to prepare anatase TiO2 (a-TiO2)
thin films on glass substrates with dimension of 10  25 mm2.
TiCl4 was added dropwise to ethanol (50 ml) with a volume ratio
of 1/10 while it was stirring. TiO2 films were obtained by dip-coat-
ing method. At first, the substrates were cleaned by abluent, deion-
ized (DI) water (18 MX) and acetone in turns. Then, they were
immersed in the sol for about 1 min and pulled up vertically at a
speed of 1 mm/s. After drying the samples at room temperature
for 24 h, they were subsequently heated at 100 °C for 1 h. The crys-
tallization of the films was occurred by heat treatment at 500 °C for
60 min. Thickness of the films was measured about 200 nm by a
profilometer. The prepared a-TiO2 thin film was finally immersed
in H2SO4 solution (0.05 M) for 5 min, for roughening its surface.
2.2. Preparation of Ag/a-TiO2 thin films
To deposit Ag nanoparticles in the prepared a-TiO2 thin film, the
film was immersed in a beaker containing AgNO3 aqueous solution
(5 mg/ml), after carefully cleaning with DI water, methanol and 2 h
UV irradiation. After drying the solutions containing the films for
24 h, the as-prepared films were washed several times with dis-
tilled water to remove the Ag ions adsorbed on the surface. Then,
they were irradiated with UV light at wavelength of 255 nm for
24 h to convert the Ag ions to Ag metal. After this stage, color of
the films was light yellow.
2.3. Preparation of Ag–TiO2/Ag/a-TiO2 thin films
The Ag–TiO2 sol was prepared by dropwise adding TiCl4 to eth-
anol (25 ml) with a volume ratio of 1/10 and then adding DI water
(25 ml) and silver nitrate (5 mg/ml AgNO3 in the solution) while
the mixture was stirring. The prepared Ag/a-TiO2 thin films were
soaked in the Ag–TiO2 aqueous solution for 5 min and pulled up
vertically at a speed of 0.2 mm/s, after cleaning with DI water,
methanol and 2 h UV irradiation. Then, they were subsequently
heated at 100 °C for 1 h to convert the Ag ions to Ag metal. Thick-
ness of the Ag–TiO2 films grown by this method on a bare glass
substrate was previously measured about 30 nm. The prepared
films were transparent with light yellow color and washed several
times with DI water to remove the Ag ions adsorbed on the surface.
It was also possible to convert the silver ions embedded in the TiO2
thin film to metallic Ag particles by using the UV irradiation (sim-
ilar to the last procedure). However, UV irradiation did not lead to a
self-accumulation of Ag nanoparticles on surface of the TiO2 thin
film, in contrast with the heat treatment-based procedure (applied
in this work) which resulted in a self-accumulation of metallic Ag
nanoparticles on the film surface.
2.4. Characterization of the thin films
Phase formation and crystalline properties of the films were
investigated by X-ray diffraction (XRD) obtained by using an X’Pert
PRO MRD (PANalytical) analyzer with a Cu Ka radiation source.
X-ray photoelectron spectroscopy (XPS) was employed to study
the surface atomic composition and chemical state of the Ag–TiO2/
Ag/a-TiO2 thin films. XPS data were obtained using a hemispherical
analyzer with an Al Ka X-ray source (hm = 1486.6 eV) operating at a
vacuum better than 107 Pa. All binding energy values were deter-
mined by calibration and fixing the C(1s) line to 285 eV. Deconvo-
lution of the XPS peaks was performed by using the SDP Ver. 4.0
software. Both survey scans and individual high-resolution scans
for Ag(3d), Ti(2p), O(1s) and C(1s) peaks were recorded. XPS depth
profile acquisitions were also acquired by Ar+ ion beam sputtering
with incident energy of 1 keV and the sputtering rate of about
0.5 nm/min.
Surface morphology of the thin films and surface distribution of
the Ag nanoparticles on the films were studied by a Philips XL30
scanning electron microscopy (SEM). Size of the silver nanoparti-
cles in the Ag–TiO2 thin film was determined by transmission elec-
tron microscopy (TEM)-Philips CM200. The TEM sample was
prepared by soaking a holey carbon coated Cu grid in the AgNO3
solution, dipping the dried grid in the Ag–TiO2 sol, and finally,
heating the dip-coated grid at 100 °C in air. The acceleration volt-
age of 200 kV was applied for the TEM observation.
A Jasco V530 UV–vis spectrophotometer was used to determine
the optical absorption of the films in the wavelength range of 300–
1100 nm with 1 nm resolution.
2.5. Antibacterial test
For antibacterial test of the a-TiO2, Ag/a-TiO2 and Ag–TiO2/Ag/a-
TiO2 thin films, Escherichia coli (E. coli, ATCC 25922) was selected as
a model for the Gram-negative bacteria. Before the microbiological
experimentation, all glass wares and samples were sterilized by
autoclaving at 120 °C for 15 min. In each run, one of the synthe-
 O. Akhavan / Journal of Colloid and Interface Science 336 (2009) 117–124 119

sized thin films was immersed in 10 ml nutrient broth inoculated

TiO 2 (101)
a)
by about 105 cfu (colony forming units)/ml of E. coli. The solutions
containing the different samples were kept in the dark or under
visible or the solar light. The visible light was obtained by a white

Intensity (arb. unit)

regular fluorescent lamp (10 W, wavelength peak at 425 nm) with
an optical high-pass filter (k > 410 nm, L41, Kenko Co.). The light

TiO 2 (200)
power was 1 mW/cm2 at the position of the catalyst. The bacte-
rial photodegradation of the photocatalysts was also checked dur-

TiO 2 (004)

TiO 2 (105)
TiO 2 (211)
ing the summer in Tehran (IRAN) at around noon under the solar

TiO 2 (204)
light irradiation. The average solar intensity was measured
1 mW/cm2. For easily and correctly counting the grown bacterial
colonies, 0.1 ml of the treated solution was diluted with DI water
to a suitable volume. Then, the diluted solution was spread on a
nutrient agar plate and incubated at 37 °C for 48 h for counting
the bacterial colonies. The reported data were the average value
of three parallel runs or the values showed the best fitting for an
exponential reduction. The dispersion of each data point was about 10 20 30 40 50 60 70
50% of that point which is not so considerable in analysis of a log- 2θ (degree)
arithmic diagram.

TiO 2 (101)
2.6. Silver ions release from the Ag–TiO2/Ag/a-TiO2 thin film
b)

For silver ion release detection, at first, 10 samples of the Ag/a-

Intensity (arb. unit)

TiO2 or the Ag–TiO2/Ag/a-TiO2 nanocomposite thin films were im-
mersed in a flask containing 10 ml HNO3 solution (0.1 M) at room

TiO 2 (200)
temperature. Then, the concentrations of Ag+ released were mea-
sured by anodic stripping voltammetry (ASV) of 1 ml of the pre-

TiO 2 (004)
+Ag(111)
pared solution, as also previously described by others [17,62].

TiO 2 (105)
TiO 2 (211)
The ASV measurements for silver ions release were carried out

TiO 2 (204)
using an Autolab PGSTAT100 Potentiostat/Galvanostat, using a

Ag(200)
conventional three-electrode electrochemical cell containing a

Ag(220)
glass carbon electrode and a Pt wire as the working and counter
electrodes, respectively. In addition, Ag/AgCl electrode with satu-
rated KCl solution was employed as reference electrode. The dis-
charge potential was 0.55 V, and the stripping scan was carried
out in the potential range of 0–0.6 V with a scan rate of 100 mV/ 10 20 30 40 50 60 70
s. It should also to be noted that the current of released silver ions
was calibrated by measuring the currents of AgNO3 solutions with 2θ (degree)
various concentrations, in our experimental conditions.
Fig. 1. XRD patterns of (a) the Ag/a-TiO2 and (b) the Ag–TiO2/Ag/a-TiO2
nanocomposite thin films.
3. Results and discussion

Fig. 1 shows XRD patterns of the synthesized Ag/a-TiO2 and Ag–
TiO2/Ag/a-TiO2 thin films. Both the patterns show diffraction peaks
relating to (1 0 1), (0 0 4), (2 0 0), (1 0 5), (2 1 1), and (2 0 4) planes of
anatase phase of TiO2 (a-TiO2) which is the favorable structure
with better photocatalytic functional properties than its rutile
phase. The mean crystalline size of the TiO2 film was calculated
11 nm, based on the peak broadening analysis described by
Scherrer’s equation for the (1 0 1) peak. Fig. 1a shows no peak re-
lated to crystallization of silver particles in the Ag/a-TiO2 thin film.
On the other hand, XRD pattern of the Ag–TiO2/Ag/a-TiO2thin film
(Fig. 1b) clearly exhibits the (2 0 0) and (2 2 0) diffraction peaks of
the metallic silver. Since these peaks are attributed to face centered
cubic (fcc) structure of metallic silver, the Ag(1 1 1) peak can be
overwhelmed in the stronger TiO2(0 0 4) peak. The mean crystalline
size of the Ag nanoparticles, determined using the Scherrer’s equa-
tion on (2 0 0) diffraction peak, was calculated about 12 nm.
Since the Ag–TiO2/Ag/a-TiO2 thin films were prepared to be ap-
plied in the antibacterial purposes, their surface analysis and so
understanding their surface properties are necessary. Fig. 2 shows
the high-resolution XPS peaks of Ti(2p), Ag(3d) and O(1s) core lev-
els of the Ag–TiO2/Ag/a-TiO2 thin film. The Ti(2p) XPS peak has
been presented in Fig. 2a. Two peaks in the Ti(2p) binding energy
region were observed. The peak located at binding energy of
464.7 eV corresponds to the Ti(2p1/2) and another one located at
459.1 eV is assigned to the Ti(2p3/2). The slitting between Ti(2p1/
2) and Ti(2p3/2) core levels is 5.6 eV, indicating a normal state of
Ti4+ in the anatase TiO2 film. High-resolution XPS spectrum of
Ag(3d) core level has been shown in Fig. 2b. The Ag(3d5/2) and
Ag(3d3/2) peaks were found at binding energy of 368.5 eV and
374.5 eV, respectively. Moreover, the slitting of the 3d doublet of
Ag is 6.0 eV, indicating formation of metallic silver nanoparticles
[63,64] on surface of the Ag–TiO2/Ag/a-TiO2 thin film. To further
study the chemical state of the silver atoms accumulated on the
surface, a detailed deconvolution of the Ag(3d) peak was also per-
formed. The binding energy of Ag(3d5/2) core level for Ag, Ag2O and
AgO is 368.6, 368.2 and 367.8 eV, respectively. Hence, the Ag(3d5/2)
peak was deconvoluted into three Gaussian components with
identical full width at half maximum (FWHM) after a Shirley back-
ground subtraction. Based on the deconvolution analysis, we have
found that about 91% of the silver atoms accumulated on the sur-
face were in the Ag0 (metallic) state, while only about 1% and 8% of
the silver atoms were in Ag+ and Ag2+ chemical states, respectively.
The O(1s) photoelectron peak of the Ag–TiO2/Ag/a-TiO2 thin film
has been presented in Fig. 2c. This peak was also deconvoluted into
three Gaussian components with identical FWHM after a Shirley
background subtraction. The first component located at 530.7 eV
120 O. Akhavan / Journal of Colloid and Interface Science 336 (2009) 117–124

368.5 eV Agº (91%)

O2- Ag+ (1 %)
OH¯ Ag2+ (8 %)
H2O Fitted peak
Fitted peak Exp. Peak
c) Exp. Peak b)
374.5 eV

6.0 eV
Intensity (arb. unit)

528 530 532 534 536 365 367 369 371 373 375 377 379

5.6 eV
a)

459.1 eV 464.7 eV

453 455 457 459 461 463 465 467 469

Binding Energy (eV)

Fig. 2. XPS spectra of (a) Ti(2p); (b) deconvolution of Ag(3d) and (c) deconvolution of O(1s) core levels of the Ag–TiO2/Ag/a-TiO2 nanocomposite thin film.

1.2
was assigned to the lattice oxygen of the TiO2 thin film. The second
component at 532.0 eV corresponds to the oxygen of surface OH
bound in –Ti(OH)–O–Ti–(OH)–, which in photo generation of elec- 1.0 Ag/TiO2
tron–hole changes into OH free radicals that is a promoting agent OH¯/TiO2
in the photocatalysis processes [2,64]. The third deconvoluted H2O/TiO2
O(1s) peak, at 533.0 eV, corresponds to oxygen of water molecules 0.8
Molar ratio

included in the film structure or adsorbed on the film surface.

The Ag/TiO2, OH/TiO2 and H2O/TiO2 molar ratios of the Ag–
0.6
TiO2/Ag/a-TiO2 thin film at the different depths of the film was cal-
culated from the area ratio of the deconvoluted XPS peaks, as
shown in Fig. 3. It is seen that the Ag/TiO2 molar ratio on the film 0.4
surface was calculated to be 0.69 which is considerably (one order
in magnitude) higher than the nominal value considered in the sol
(0.07). This indicates accumulation of the silver atoms on surface of 0.2
the TiO2 thin films, which can be an advantage in processes in
which interaction with surface Ag nanoparticles are important. A
0.0
similar observation was recently reported for self-accumulation
0 10 20 30 40 50
of silver nanoparticles on surface of aqueous silica thin film [61].
Here, we also measured the Ag/TiO2 ratio at the different depths Depth (nm)
of the thin film. The slight increase in the Ag/TiO2 ratio at the depth
Fig. 3. Variation of Ag/TiO2, OH/TiO2 and H2O/TiO2 molar ratios of the Ag–TiO2/Ag/
of 5 nm shows that more silver nanoparticles were embedded at a-TiO2 nanocomposite thin film calculated from the XPS analysis at the different
near the surface of the TiO2 layer. It was found that value of the sputtering depths.
Ag/TiO2 ratio substantially decreased, as the sputtering depth in-
creased up to 20 nm, indicating accumulation of the silver nano-
particles on the surface (from surface down to 10 nm). At depth was observed. The variation of OH/TiO2 molar ratio, calculated
of 30 nm (corresponding to the interface of Ag–TiO2 and Ag/a- from the area ratio of the second and first deconvoluted peaks of
TiO2 layers), a peak for the Ag/TiO2 ratio was observed which can the O(1s) core level, has also been presented in Fig. 3. A high value
be attributed to accumulation of the Ag particles at the interface. molar ratio of the OH/TiO2 (1.17) was observed on the film sur-
This accumulation at the interface (depth of 30 nm) can act as an face, due to more photocatalytic activity of the film surface. By
auxiliary and lasting storage of silver atoms, especially during a sil- increasing the sputtering depth, the OH/TiO2 molar ratio substan-
ver ion releasing process. By increasing the depth of the thin film tially decreased to 0.06 at 25 nm. But, at 30 nm (location of the
from 30 to 45 nm, a considerable decrease in the Ag/TiO2 ratio interface), the OH/TiO2 molar ratio showed a small peak. This
 O. Akhavan / Journal of Colloid and Interface Science 336 (2009) 117–124 121

peak indicated further presence of the aqueous pores at the bound-

ary surfaces of the Ag–TiO2 and Ag/a-TiO2 layers in the Ag–TiO2/ c)
Ag/a-TiO2 thin film. In addition, the possibility in production of
OH bound at both the interface and film surface (where the silver
atoms accumulated) can cause the released silver atoms and/or

Absorption (arb. unit)

particles are in their positive ionic state (not in the metallic state).
The H2O/TiO2 molar ratio of the film at the different depths was
also calculated from the area ratio of the third and first deconvo-
luted peaks of the O(1s) core level. From Fig. 3 it is seen that by
increasing the depth, the water ratio decreased. This means that
surface of the Ag–TiO2/Ag/a-TiO2 thin film was still aqueous and
so porous, which can be an advantage in surface interactions and
controlling the silver ion releasing processes. The weak peak lo-
cated at 30 nm, shows that a relative more water can be found at
the interface due to presence of further pores and/or grooves at
there, in consistency with the observed small peak for the OH/
TiO2 molar ratio. It should also be noted the water included in
the film is cause of the out-diffusion of the metallic nanoparticles,
initially dispersed in the film, into the surface, during a low tem-
perature (100 °C) annealing process, as previously indicated else-
where [61].
Surface morphology of the Ag–TiO2/Ag/a-TiO2 thin film was
studied by SEM, as shown in Fig. 4. The XPS analysis showed that
the silver particles were accumulated on the surface. Thus, the par-
ticles observed on the surface were attributed to the Ag nanoparti-
cles on the surface of aqueous TiO2 film. The diameter histogram of
the surface particles (shown as the bottom inset of Fig. 4) indicated
that average diameter of these surface nanoparticles are about
35 nm. TEM image of the Ag nanoparticles in the Ag–TiO2 film
has also been presented as the top inset of Fig. 4. It shows the silver
nanoparticles with diameters in the range of 13–31 nm.
The UV–vis measurement of the Ag/a-TiO2 and Ag–TiO2/Ag/a-
TiO2 thin films exhibited a single surface plasmon resonance
(SPR) absorption peak centered at 412 nm, as shown in Fig. 5. It
is well-known that, the UV–vis absorption spectra of the semi-
spherical Ag nanoparticles with sizes ranged from 3 to 20 nm exhi-
bit a single absorption peak at 410 nm [65–67]. Therefore,
consistent with the XPS results, SPR band showed that the silver-
based nanoparticles in the Ag/a-TiO2 and also in the air-annealed
Ag–TiO2/Ag/a-TiO2 thin films were metallic silver nanoparticles.
The narrower and lower intensity SPR absorption peak of the Ag/
a-TiO2 thin film than the Ag–TiO2/Ag/a-TiO2 can be due to smaller
b)
a)
300 500 700 900 1100
Wavelength (nm)
Fig. 5. UV–vis absorption spectra of (a) the a-TiO2, (b) the Ag/a-TiO2, and (c) the Ag–
TiO2/Ag/a-TiO2 photocatalyst thin films.
size and lower concentration of the silver nanoparticles in the for-
mer thin film structure, respectively. For a better comparison, the
optical absorption spectrum of the a-TiO2 thin film was also shown
in Fig. 5, indicating the transparency of the synthesized a-TiO2 thin
film.
The bactericidal activity of the a-TiO2, Ag/a-TiO2 and Ag–TiO2/
Ag/a-TiO2 thin films, as compared to a blank glass substrate (con-
trol sample), were studied against E. coli bacteria in dark and in
the visible and the solar light irradiations, as shown in Fig. 6. It is
seen that the viable bacteria reduced in an exponential manner.
The slope of the fitted line yields relative rate of reduction of the
number of viable bacteria. The a-TiO2 thin films showed a weak
antibacterial activity with the relative rate of reduction of about
0.7  102 min1 in dark. The photo-antibacterial activity of the
a-TiO2 thin films was also checked in the visible and solar light
irradiations. No change was observed for antibacterial activity of
the a-TiO2 thin films in the visible light, as compared to its activity
in dark. But, the a-TiO2 improved the rate of reduction of the viable

Fig. 4. SEM image of the Ag–TiO2/Ag/a-TiO2 nanocomposite thin film. The bottom inset shows diameter histogram of the particles accumulated on the surface. The top inset
shows TEM image of Ag nanoparticles embedded in the TiO2 thin film.
122 O. Akhavan / Journal of Colloid and Interface Science 336 (2009) 117–124
1000000 Blank samples
100000
Cell forming unit (cell/ml)
a-TiO2
10000
1000
100
Ag/a-TiO2
Ag-TiO2/Ag/a-TiO2
10
1 silver ions to 80% of its saturation amount in the solution at a long
0 20
0 40 60 80
0 100 120 140 160 180 200
Time (min)
Fig. 6. CFU of E. coli cultured for various periods in a medium containing the a-TiO2,
Ag/a-TiO2 and Ag–TiO2/Ag/a-TiO2 photocatalyst thin films as compared to blank
(control) sample, in dark (j) and under irradiation of the visible light (N) and the
solar light (d).
bacteria up to about 20% in the solar light irradiation. By deposition
of the Ag nanoparticles on the a-TiO2 thin film (the Ag/a-TiO2 thin
film), the rate of reduction of viable bacteria highly increased to
2.7  102 min1 corresponding to 190 min for killing all the bac-
teria in dark. The photocatalytic activity of the Ag/a-TiO2 thin films
was also checked in the visible and the solar light irradiations. In
the visible light, the relative rate of reduction of the viable bacteria
was found 3.4  102 min1 corresponding to 150 min to kill all
the bacteria, indicating an improvement than the dark condition.
This improvement showed that the Ag/a-TiO2 thin film can also
work better in the solar light irradiation, due to its UV spectrum.
In fact, it was observed that the antibacterial activity of the Ag/a-
TiO2 thin films was improved in the solar light irradiation with
the rate of reduction of 4.3  102 min1 (60% improvement than
the dark condition) and the required time of 120 min to kill all the
E. coli bacteria. By adding the Ag–TiO2 nanocomposite layer onto
the Ag/a-TiO2 thin film, the rate of reduction of viable bacteria in-
creased to 3.6  102 min1 corresponding to 140 min for killing
all the bacteria in dark. For the Ag–TiO2/Ag/a-TiO2 thin film, the
relative rate of reduction of the viable bacteria was improved to
4.7  102 and 5.8  102 min1 corresponding to 110 and
90 min for killing all the bacteria, in the visible and solar light irra-
diations, respectively. Therefore, the Ag–TiO2/Ag/a-TiO2 thin film
structure can efficiently improve the photocatalytic activity of
TiO2 and Ag doped TiO2 thin films in dark and in the solar light
irradiation.
It was recently reported that bactericidal efficiency of silver ions
in AgNO3 can significantly be enhanced by UV-A or visible light
irradiation [68,69]. Hence, we also checked the effect of light irra-
diation of a mercury lamp (peak wavelengths at 275, 350 and
660 nm) on the bactericidal efficiency of Ag nanoparticles in an
Ag–SiO2 thin film prepared similar to the Ag–TiO2 thin film used
at this work. Details of the Ag–SiO2 thin film preparation can be
found elsewhere [61,66]. Anyway, no considerable change in the
antibacterial activity of the Ag–SiO2 thin film irradiated by the light
was observed as compared to its activity in dark (of course, after
elimination of the irradiation effect for a blank sample). Therefore,
the silver nanoparticles in the Ag–SiO2 thin film and also in the Ag–
TiO2/Ag/a-TiO2 thin film structure did not any contribution in the
photodegradation (not in the usual degradation) of the bacteria.
This result might also be expected from the XRD pattern of the
Ag–TiO2/Ag/a-TiO2 thin film (Fig. 1b) which shows no peak related
to the crystalline AgNO3, and from its XPS analysis (Fig. 2b) and
optical absorption spectrum (Fig. 5c) which show formation of
metallic silver nanoparticles, in contrast to the results shown by
Liu et al. [69] and Kim et al. [68] who showed formation of silver
ions (not silver metals) and their role in the photodegradation of
bacteria, respectively.
The silver ion releasing of the Ag/a-TiO2 and Ag–TiO2/Ag/a-TiO2
thin films was also investigated. Fig. 7 shows the amounts of the
released silver ions from the Ag/a-TiO2 (Fig. 7a) and Ag–TiO2/Ag/
a-TiO2 (Fig. 7b) thin films as a function of the elapsing time in
the HNO3 solution, after three sequent measurements. For the
Ag/a-TiO2 thin film, it is seen that silver ions were initially released
with a high rate in the first five days. This time can be called satu-
ration time defined as the time required for reaching the amount of
time. After the saturation time, however, the process of releasing
silver ions was controlled in a relatively slower release rate. The
initial stage of silver ion release process is generally controlled
by the water diffusion on surface of the matrix. For both the Ag/
a-TiO2 and the Ag–TiO2/Ag/a-TiO2 thin films with silver nanoparti-
cles accumulated on the surface, easy water diffusion on the sur-
face could be resulted in a sharp release of the silver ions at the
initial stage. On the other hand, the considerable reduction in the
release rate of silver ions at the latter stage (the saturation stage)
can be attributed to the slow diffusion of water in the pores of
the TiO2 film, and so change in the silver ion releasing mechanism.
This change in the release behavior of the silver ions from both the
Ag/a-TiO2 and the Ag–TiO2/Ag/a-TiO2 thin films completely distin-
guishes them from the bulk silver-based materials exhibiting a
sharp ion release after a minimum releasing at initial stages [62].
Moreover, it was found that after the three sequent measurements
on the Ag/a-TiO2 thin film, the saturation amount of the silver ions
in the solution reduced from 9.4 to 4.0 nM/ml, corresponding to
57% reduction in the initial saturation amount. This indicates a
high consumption of the accumulated silvers on surface of the a-
10 1st Measurement
9 tsat = 5 days
Concentration (μM/ml)
15
8 10
Ag+ release (nM/ml)
5
7 0 a)
0 10 20 30 40 50 60
6 Current (μA)
5
4 3rd Measurement
3 1st Measurement
2
3rd Measurement b)
1 t sat = 8 days
0
0 5 10 15 20 25
Time (day)
Fig. 7. The silver ion release curves of (a) the Ag/a-TiO2 and (b) the Ag–TiO2/Ag/a-
TiO2 nanocomposite thin films, after three sequent measurements in HNO3
solution. The inset shows the obtained standard curve of silver ions for calibration
of the ion current.
 O. Akhavan / Journal of Colloid and Interface Science 336 (2009) 117–124 123

TiO2 thin film. For the Ag/a-TiO2 thin film covered by the Ag–TiO2 Ag–TiO2/Ag/a-TiO2 photocatalyst is an excellent and long lasting
nanocomposite layer (i.e., Ag–TiO2/Ag/a-TiO2 nanocomposite thin antibacterial nanocomposite material appropriate for future bio-
film), however, the situation is completely different. Fig. 7b shows medical and bactericidal applications.
that, this time, the saturation time is eight days which means a
slower silver ion release rate at the first stage and consequently Acknowledgments
a lower saturation amount of the silver ions in the solution, as
compared to the obtained characteristics of the Ag/a-TiO2. The The authors would like to thank Research Council of Sharif Uni-
reduction observed in the release rate of the Ag–TiO2/Ag/a-TiO2 versity of Technology and also Iranian Nanotechnology Initiative
thin film also confirms that the silver particles accumulated on for financial support of the project.
the film surface were effectively protected by a very thin ( nm)
titania layer (as also resulted based on the depth profile XPS anal- References
ysis) which controls further the releasing process. In addition, it
was observed that even after the three sequent ion release mea- [1]
[2]
surements, the saturation amount only decreased from 2.8 to [3]
2.5 nM/ml, corresponding to only 10% reduction in the initial satu- [4]
ration amount. This means that by covering the Ag/a-TiO2 thin film [5]
[6]
with the Ag–TiO2 nanocomposite (barrier) layer, further control- [7]
ling the silver ion release is provided so that durability of the [8]
Ag–TiO2/Ag/a-TiO2 nanocomposite thin film (of course with higher [9]
(about 2 times) silver storage) increases to at least 11 times of the [10]
Ag/a-TiO2 thin film. Therefore, the Ag–TiO2/Ag/a-TiO2 thin film can [11]
be considered as efficient and also lasting antibacterial nanocom-
[12]
posite materials for future antibacterial and bio-medical
applications. [13]
[14]
[15]
4. Summary [16]
[17]
The Ag–TiO2/Ag/a-TiO2 nanocomposite thin film photocatalyst [18]
sensitive to the solar light with an efficient storage of Ag nanopar- [19]
ticles both on the film surface and at interface of the Ag–TiO2 and [20]
a-TiO2 thin films was simply synthesized by sol–gel. XRD analysis [21]
[22]
confirmed formation of a-TiO2 and crystalline metallic silver grains [23]
with fcc structure. The SPR absorption band at 412 nm in wave- [24]
length was attributed to formation of spherical Ag nanoparticles
[25]
in the Ag doped thin films. The depth profile XPS analysis showed
that the Ag nanoparticles (average diameter of 35 nm) were [26]
effectively embedded at depths of 5 nm of the aqueous TiO2 layer [27]
and at depth of 30 nm (at the interface) of the nanocomposite film.
[28]
In addition, the OH bounds and also water contents of the film [29]
were concentrated both on the film surface and at the interface, [30]
as an advantage for release of more ionic (not metallic) silver nano- [31]
[32]
particles. The photodegradation of the thin films was studied by [33]
testing its antibacterial activity against E. coli bacteria, in dark
and in the visible and the solar light irradiations. The relative rate [34]
of reduction of the viable bacteria for the nanocomposite film [35]
(3.6  102 min1) was 5.1 times greater than the corresponding [36]
value for the a-TiO2 thin film, in dark. In the solar light irradiation,
[37]
the rate of reduction of the viable bacteria for the Ag–TiO2/Ag/a- [38]
TiO2 film (5.8  102 min1) was measured 1.35 and 6.90 times
greater than the corresponding rate for the Ag/a-TiO2 and a-TiO2 [39]
thin films, respectively. The ion release measurement indicated
[40]
that the Ag/a-TiO2 and Ag–TiO2/Ag/a-TiO2 thin films show a satu- [41]
ration behavior at long times (5 and 8 days in HNO3 solution,
respectively), which is a completely different behavior from the [42]
general behavior of ion release in silver-based bulk materials. [43]
Hence, the ion releasing process was mainly controlled by inter-
diffusion of water and silver nanoparticles through the pores of [44]
[45]
the films, not by easy surface diffusion of water on the Ag nanopar-
ticles. Consistent with the XPS result, the ion release measure- [46]
ments also showed the surface Ag nanoparticles in the Ag–TiO2/
[47]
Ag/a-TiO2 film were protected by a nanometric TiO2 barrier layer [48]
against an easy ion release. Three sequent ion release measure-
ments of the Ag/a-TiO2 and Ag–TiO2/Ag/a-TiO2 films showed that [49]
[50]
durability of the latter film (of course with 2 times silver storage)
is at least 11 times greater than the former one. Therefore, the [51]
A. Fujishima, K. Honda, Nature 238 (1972) 37.
M.S. Hoffmann, T. Martin, W. Choi, D.W. Bahnemann, Chem. Rev. 95 (1995) 69.
G.A. Somorjai, Chem. Rev. 96 (1996) 1223.
A. Mills, S.L. Hunte, J. Photochem. Photobiol. A 108 (1997) 1.
C. Burda, X. Chen, R. Narayanan, M.A. El-Sayed, Chem. Rev. 105 (2005) 1025.
R. Hoffmann, T. Martin, Y. Choi, W. Bahnemann, Chem. Rev. 95 (1995) 69.
J. Robertson, P. Robertson, L. Lawton, J. Photochem. Photobiol. A 175 (2005) 51.
J. Carey, J. Lawrence, H. Tosine, Bull. Environ. Contam. Toxicol. 16 (1976) 697.
V. Nadtochenko, A. Rincon, S. Stanca, J. Kiwi, J. Photochem. Photobiol. A 169
(2005) 131.
A. Fujishima, N.R. Tata, A. Tryk, J. Photochem. Photobiol. C 1 (2000) 1.
T. Matsunaga, R. Tomoda, T. Nakajima, H. Wake, FEMS Microbiol. Lett. 29
(1985) 211.
V. Nadtochenko, N. Denisov, O. Sarjusiv, D. Gumy, C. Pulgarin, J. Kiwi, J.
Photochem. Photobiol. A 181 (2006) 401.
G. Rincon, C. Pulgarin, Appl. Catal. B 51 (2004) 283.
J.C. Yu, W. Ho, J. Lin, H. Yip, P.K. Wong, Environ. Sci. Technol. 37 (2003) 2296.
J.S. Hur, Y. Koh, Biotechnol. Lett. 24 (2002) 23.
K. Sunada, T. Watanabe, K. Hashimoto, Environ. Sci. Technol. 37 (2003) 4785.
Y. Liu, X. Wang, F. Yang, X. Yang, Micropor. Mesopor. Mater. 114 (2008) 431.
M. Bellantone, H.D. Williams, L.L. Hench, Antimicrob. Agents Chemother. 16
(2002) 1940.
A. Henglein, J. Phys. Chem. 97 (1993) 5457.
A.P. Alivisatos, J. Phys. Chem. 100 (1996) 13226.
Y. Murakami, J. Matsumoto, Y. Takasu, J. Phys. Chem. B 103 (1999) 1836.
G. Fu, P.S. Vary, C.T. Lin, J. Phys. Chem. B 109 (2005) 8889.
W.O. Williamson, Nature 143 (1939) 279.
M. Anpo, S. Kishiguchi, Y. Ichihashi, M. Takeuchi, H. Yamashita, K. Ikeue, B.
Morin, A. Davidson, M. Che, Res. Chem. Intermediates 27 (2001) 459.
J.C. Yu, W. Ho, J. Yu, H. Yip, P.K. Wong, J. Zhao, Environ. Sci. Technol. 39 (2005)
1175.
R. Asahi, T. Morikawa, T. Ohwaki, K. Aoki, Y. Taga, Science 293 (2001) 269.
C. Burda, Y. Lou, X. Chen, A.C.S. Samia, J. Stout, J.L. Gole, Nano Lett. 3 (2003)
1049.
S.U.M. Khan, M. Al-Shahry, W.B. Ingler Jr., Science 297 (2002) 2243.
C.H. Park, S.B. Zhang, S.H. Wei, Phys. Rev. B 66 (2002) 073202.
W. Choi, A. Termin, M. Hoffmann, J. Phys. Chem. 98 (1994) 13669.
W. Mu, J.M. Herrmann, P. Pichat, Catal. Lett. 3 (1989) 73.
D. Duonghong, E. Borgarello, M. Gratzel, J. Am. Chem. Soc. 103 (1981) 4685.
J. Husheng, H. Wensheng, W. Liqiao, X. Bingshe, L. Xuguang, Dent. Mater. 24
(2008) 244.
X. Bingshe, N. Mei, W. Liqiao, H. Wensheng, L. Xuguang, J. Photochem.
Photobiol. A 188 (2007) 98.
Q. Cheng, C. Li, V. Pavlinek, P. Saha, H. Wang, Appl. Surf. Sci. 252 (2006) 4154.
S.-D. Oh, S. Lee, S.-H. Choi, I.-S. Lee, Y.-M. Lee, J.-H. Chun, H.-J. Park, Colloids
Surf. A 275 (2006) 228.
O. Akhavan, E. Ghaderi, Sci. Technol. Adv. Mater. 10 (2009) 015003.
T. Yuranova, A.G. Rincon, A. Bozzi, S. Parra, J. Photochem. Photobiol. A 161
(2003) 27.
Q.L. Feng, J. Wu, G.Q. Chen, F.Z. Cui, T.N. Kim, J.O. Kim, J. Biomed. Mater. Res. 52
(2000) 662.
I. Sondi, B. Salopek-Sondi, J. Colloid Interface Sci. 275 (2004) 177.
Y. Matsumura, K. Yoshikata, S. Kunisaki, T. Tsuchido, Appl. Environ. Microbiol.
69 (2003).
X.H. Xu, W.J. Brownlow, S.V. Kyriacou, Q. Wan, J.J. Viola, Biochemistry 43
(2004) 10400.
S.K. Gogoi, P. Gopinath, A. Paul, A. Ramesh, S.S. Ghosh, A. Chattopadhyay,
Langmuir 22 (2006) 9322.
S.V. Kyriacou, W.J. Brownlow, X.H. Xu, Biochemistry 43 (2004) 140.
M.K. Seery, R. George, P. Floris, S.C. Pillai, J. Photochem. Photobiol. A 189 (2007)
258.
K. Page, R.G. Palgrave, I.P. Parkin, M. Wilson, S.L.P. Savin, A.V. Chadwick, J.
Mater. Chem. 17 (2007) 95.
M.P. Reddy, A. Venugopal, M. Subrahmanyam, Water Res. 41 (2007) 379.
E.V. Skorb, L.I. Antonouskaya, N.A. Belyasova, D.G. Shchukin, H. Möhwald, D.V.
Sviridov, Appl. Catal. B 84 (2008) 94.
C. Hu, Y. Lan, J. Qu, X. Hu, A. Wang, J. Phys. Chem. B 110 (2006) 4066.
H.M. Sung-Suh, J.R. Choi, H.J. Hah, S.M. Koo, Y.C. Bae, J. Photochem. Photobiol. A
163 (2004) 37.
A. Sclafani, M.N. Mozzanega, J.M. Herrmann, J. Catal. 168 (1997) 117.
124 O. Akhavan / Journal of Colloid and Interface Science 336 (2009) 117–124
[52] Y. Tian, T. Tatsuma, Chem. Commun. 16 (2004) 1810.
[53] Y.L. Wang, Y.Z. Wan, X.H. Dong, G.X. Cheng, H.M. Tao, T.Y. Wen, Carbon 36
(1998) 1567.
[54] H.J. Lee, S.Y. Yeo, S.H. Jeong, J. Mater. Sci. 38 (2003) 2199.
[55] C.N. Lok, C.M. Ho, R. Chen, Q.Y. He, W.Y. Yu, H. Sun, P.K.H. Tam, J.F. Chiu, C.M.
Che, J. Proteome. Res. 5 (2006) 916.
[56] J.R. Morones, J.L. Elechiguerra, A. Camacho, K. Holt, J.B. Kouri, J.T. Ramirez, M.J.
Yacaman, Nanotechnology 16 (2005) 2346.
[57] T. Maneerung, S. Tokura, R. Rujiravanit, Carbohydr. Polym. 72 (2008) 43.
[58] S.S. Mahapatra, N. Karak, Mater. Chem. Phys. 112 (2008) 1114.
[59] S. Zhang, R. Fu, D. Wu, W. Xu, Q. Ye, Z. Chen, Carbon 42 (2004) 3209.
[60] Y.Z. Wan, Y.L. Wang, G.X. Cheng, H.L. Luo, X.H. Dong, Carbon 39 (2001) 1607.
[61] O. Akhavan, R. Azimirad, A.Z. Moshfegh, J. Phys. D 41 (2008) 195305.
[62] R. Kumar, H. Munstedt, Biomaterials 26 (2005) 2081.
[63] J.F. Moulder, W.F. Stickle, P.E. Sobol, K.D. Bomben (Eds.), Hand Book of X-ray
Photoelectron Spectroscopy, Perkin–Elmer Corporation Physical Electronics
Division, 1992.
[64] C.D. Wagner, A.V. Naumkin, A. Kraut-Vass, J.W. Allison, C.J. Powell, J.R. Rumble
Jr., NIST Standard Reference Database 20, Ver. 3.4, 2006. Available from:
<http://srdata.nist.gov/xps/index.htm>.
[65] W. Li, S. Seal, E. Megan, J. Ramsdell, K. Scammon, G. Lelong, L. Lachal, K.A.
Richardson, J. Appl. Phys. 93 (2003) 9553.
[66] A. Babapour, O. Akhavan, R. Azimirad, A.Z. Moshfegh, Nanotechnology 17
(2006) 763.
[67] H. Jia, W. Xu, J. An, D. Li, B. Zhao, Spectrochim. Acta A 64 (2006)
956.
[68] J.Y. Kim, C. Lee, M. Cho, J. Yoon, Water Res. 42 (2008) 356.
[69] T.-X. Liu, X.-Z. Li, F.-B. Li, Environ. Sci. Technol. 42 (2008) 4540.