Carbohydrate Polymers 169 (2017) 101–107

Contents lists available at ScienceDirect

Carbohydrate Polymers
journal homepage: www.elsevier.com/locate/carbpol

Preparation of chitosan-TiO2 composite film with efficient

antimicrobial activities under visible light for food packaging
applications
Xiaodong Zhang, Gang Xiao, Yaoqiang Wang, Yan Zhao, Haijia Su ∗ , Tianwei Tan
State Key Laboratory of Chemical Resource Engineering, Beijing Advanced Innovation Center for Soft Matter Science and Engineering, Beijing University of
Chemical Technology, Beijing 100029, PR China

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

Article history: By incorporation of TiO2 nano-powder in chitosan, a chitosan-TiO2 composite film was prepared with effi-
Received 22 December 2016 cient antimicrobial activity against food-borne pathogenic microbes and expected to be a promising food
Received in revised form 2 March 2017 packaging material. Scanning electron microscopy analysis showed that the TiO2 nano-powder was suc-
Accepted 23 March 2017
cessfully and uniformly dispersed into the chitosan matrix. TiO2 addition led to enhanced hydrophilicity,
Available online 25 March 2017
to better mechanical properties, and to decreased light transmittance in visible light region of the com-
posite film. The chitosan-TiO2 film possessed efficient antimicrobial activity against four tested strains,
Keywords:
i.e. Escherichia coli, Staphylococcus aureus, Candida albicans, and Aspergillus niger with 100% sterilization in
Chitosan
Nano-TiO2 12 h. It moreover provoked the leakage of cellular substances through damaged membrane. The prepared
Antimicrobial activity chitosan-TiO2 film was tested for packaging red grapes to prevent microbial infection and extend their
Visible light shelf life. Results were positive, stressing the potential of the novel bio-nano composite film for food
Food packaging packaging applications.
© 2017 Elsevier Ltd. All rights reserved.

1. Introduction
Recently, antibacterial and antifungal packaging, as a represen-
tative form of active packaging, has attracted increasing attention
in protecting food from food-borne microbial outbreaks and in
extending the shelf-life (Alkan et al., 2011; Tan, Lim, Tay, Lee, &
Thian, 2015). Natural materials with antimicrobial activities such
as polysaccharides, proteins, and lipids, have been proven to be
promising for food packaging applications because of their environ-
mentally friendly and biodegradable properties (Atarés & Chiralt,
2016; Chen et al., 2016; Leceta, Etxabide, Cabezudo, de La Caba,
& Guerrero, 2014; Song, Lee, Al Mijan, & Song, 2014; Tajik et al.,
2013). Chitosan is a liner cationic polysaccharide derived from
chitin (the second most abundant polysaccharide after cellulose)
(Xiao et al., 2015) by partly de-acetylation. It is one of the most
widely used active packaging materials because of its biocompati-
bility, biodegradability, excellent film forming ability, and inherent
antibacterial and antifungal properties (Bie et al., 2013; Hu et al.,
2016; Tan et al., 2015). And it is classified as a safe food preservative
∗ Corresponding author at: College of Life Science and Technology, PO Box 53,
Beijing University of Chemical Technology, Beijing 100029, PR China.
E-mail address: suhj@mail.buct.edu.cn (H. Su).
in the US, EU, and China (Al-Naamania, Dobretsova, & Dutta, 2016;
Zemljič, Tkavca, Vesel, & Šauperl, 2013). However, the mechanical
properties and antimicrobial activities of pure chitosan film were
not satisfactory for food packaging applications (Aljawish et al.,
2016; Sadeghi & Shahedi, 2016).
The incorporation into biopolymers of inorganic nano-sized
materials, such as nano-SiO2 , nano-TiO2 , and nano- montmoril-
lonite, has been proven to be an effective strategy to improve their
mechanical properties (Saha et al., 2016; Yang et al., 2016; Zhang
et al., 2016). Titanium dioxide (TiO2 ) is an attractive inorganic
nanomaterial which has raised great interest in environmental
and energy fields (Lee & Park, 2013) because of its low cost, high
photocatalytic performance, high chemical stability, and biocom-
patibility (Ma et al., 2016). The addition of TiO2 nano-powder
has been reported to enhance the mechanical properties of the
chitosan-based nanocomposite films (Mallakpour & Madani, 2015;
Yun, Yun, Yoon, & Byun, 2016).
TiO2 furthermore possesses antimicrobial capability because it
will mediate photocatalysis under light with enough energy to pro-
duce active oxygen species (ROS), such as • OH, H2 O2 , and O2 −• ,
which play a key role in destroying microbial cells (Wang, Wu,
Yang, Xue, & Liu, 2016). TiO2 photocatalysis has been widely used
for the removal of various pathogenic microorganisms containing
bacteria, fungi, and even viruses (Zhang, Liu, Li, Zhang, & Shang,

http://dx.doi.org/10.1016/j.carbpol.2017.03.073
0144-8617/© 2017 Elsevier Ltd. All rights reserved.
102 X. Zhang et al. / Carbohydrate Polymers 169 (2017) 101–107
2013). Matsunaga, Tomoda, Nakajima, and Wake (1985) reported
for the first time in 1985 that microbial cells of Lactobacillus aci-
dophilus, Saccharomyces cerevisiae, and Escherichia coli could be
killed by platinized TiO2 under near-UV light irradiation. However,
the photocatalytic antimicrobial reactions mediated by TiO2 (band
gap energy of 3.2 eV) (Verbruggen et al., 2014) can only be activated
by UV light with wavelength number shorter than 388 nm whose
energy accounts for less than 5% of the total solar irradiance on the
earth’s surface (Kuo, Tseng, Huang, & Li, 2007). Incorporating TiO2
nano powders into chitosan could extend the absorption band over
the visible light spectrum and the prepared chitosan-TiO2 com-
posite supported by cotton fibers showed synergistically enhanced
antimicrobial activities under visible light irradiation (Qian, Su, &
Tan, 2011). Recently, much attention has been focused on chitosan-
TiO2 nanocomposites which were proven to possess favorable
mechanical, thermal, and photocatalytic properties (Mallakpour
& Madani, 2015; Yun et al., 2016) and to be promising materials
in several fields, such as the removal of environmental pollutants
through adsorption and photocatalysis (Kamal, Anwar, Khan, Chani,
& Asiri, 2016), food preservation (Xu et al., 2017), coating materials
(Qian et al., 2011; Xiao, Zhang, Zhao, Su, & Tan, 2014), controlled
drug-delivery systems (Kamari & Ghiaci, 2016), wound repairing
(Fan et al., 2016), biosensors (Chang et al., 2017), etc.
In the present work, TiO2 nano-powder was introduced into
the chitosan matrix and the chitosan-TiO2 composite film was
prepared as an active antimicrobial packaging material for food
preservation. The effects of TiO2 addition on the film proper-
ties including morphology, wettability, light transmittance, and
mechanical properties were studied through the characterization
of the pure chitosan and the chitosan-TiO2 films. The antimicro-
bial activity of the prepared chitosan-TiO2 film under visible light
irradiation was tested against four typical food-borne pathogenic
microbes, i.e. E. coli, S. aureus, C. albicans, and A. niger using the
colony counting method and cellular substances leakage assays.
The realistic application of the prepared chitsan-TiO2 film in food
preservation was preliminarily evaluated for red grape packaging.
2. Materials and methods
2.1. Materials
Chitosan with 90% degree of deacetylation was extracted
from shrimp shells in our laboratory. Nano-sized TiO2 with
particle size of 50–80 nm was provided by the University
of Science and Technology Beijing (China) (Ma, Wei, Liu, &
Cao, 2009). Acetic acid, epichlorohydrin, NaOH, NaCl, NaNO3 ,
K2 HPO4 , MgSO4 , CaCl2 , phenol, and chloroform were of analyt-
ical grade and purchased from Beijing Chemical Works (China).
Tris(hydroxymethyl)aminomethane, p-nitrophenylphosphate dis-
odium salt hexahydrate, and D-glucose-6-phosphate disodium
salt hydrate were of reagent grade and purchased from Sigma-
Aldrich. Escherichia coli (E. coli, ATCC 8099), Staphylococcus aureus
(S. aureus, ATCC 6538), Candida albicans (C. albicans, ATCC 10231),
and Aspergillus niger (A. niger, ATCC 16404) were obtained from the
American Type Culture Collection. Beef peptone, beef extract pow-
der, yeast extract powder, glucose, sucrose, and agar were used for
microbial culture and purchased from Beijing AoBoXing Bio-Tech
Co., Ltd. (China). Deionized water was used in all the experiments,
except for microbial culture.
2.2. Preparation of chitosan and chitosan-TiO2 films
0.5 g chitosan and 0.05 g TiO2 nano-powder were added into
20 mL acetic acid (2.5%, v/v) and dispersed using 120 W ultrasonic
treatment for 10 min. Then the mixture was magnetically stirred
until a uniform emulsion was obtained, followed by the addition of
the cross-linker, 0.1 mL epichlorohydrin, and stirring for 4 h at room
temperature to conduct the cross-linking reaction. The emulsion
was then spread on a square glass plate (15 cm × 15 cm) which had
been coated with a layer of plastic film first and then dried naturally
overnight. The obtained chitosan-TiO2 film could be peeled off the
plate conveniently and was then soaked into 1 mol/L NaOH solution
for 2 h. Finally, the film was washed to a neutral pH with deionized
water for several times and dried at room temperature overnight.
The pure chitosan film was prepared with the same procedure
described above but without the addition of TiO2 nano-powder. The
pure chitosan films are transparent and after the addition of TiO2
nano-powder, the prepared chitosan-TiO2 films show increased
whiteness and decreased transparency which is consistent with the
previous work of Zhang et al. (2016).
2.3. Characterization
The surface and cross-section morphologies of pure chitosan
and chitosan-TiO2 films were studied by field emission scanning
electron microscopy (FE-SEM) (Hitachi S4700, Japan) with acceler-
ating voltage of 10 kV. The samples were coated by spraying gold
under vacuum before SEM evaluation. The samples were treated
with liquid nitrogen to obtain smooth cross-sections. SEM images
were gathered at magnifications of ×5000 for surface and ×500 for
cross-section analyses.
The mechanical properties of the prepared pure chitosan and
chitosan-TiO2 films were evaluated using an electronic universal
testing machine (UTM 2502, Shenzhen SUNS Technology, China).
Five strips (60 mm × 20 mm) were tested for each film. All the
measurements were conducted at the same operating parameters:
initial grip separation of 40 mm, drawing speed of 1 mm/s. The
temperature was 20 ◦ C and the relative humidity was 50%. Tensile
strength (TS, MPa) and elongation at break (E, %) were automatically
calculated and recorded by the testing system.
The contact angle (Â) for the pure chitosan and chitosan-TiO2
films were determined to evaluate the wettability using a static
water contact angle instrument (POWEREACH JC2000D3, China).
A drop of ultrapure water (5 ␮L) was placed on the surface of each
film (20 mm × 60 mm) using a micro-syringe at ambient conditions
(temperature of 20 ◦ C, relative humidity of 50%). After 15 s of expo-
sure, the drop images were recorded by a CCD camera and conveyed
to the computer for the measurement. For each film, five different
positions were tested and the average values were reported.
A UV–vis spectrophotometer (Varian Cary100, USA) was
employed to analyze the light transmittance properties of the pure
chitosan and chitosan-TiO2 films, as well as of TiO2 nanoparticles.
Their UV–vis transmittance spectra were recorded in the wave-
length range of 300–800 nm with resolution of 0.5 nm.
2.4. Antimicrobial activity
The antimicrobial activity of the prepared chitosan-TiO2 film
was evaluated employing four different pathogenic microbes as
test strains: E. coli, S. aureus, C. albicans, and A. niger. The antimi-
crobial assay was as follows: 0.1 mL of microbial cell suspension
(∼106 CFU/mL) was spread on the surface of chitosan-TiO2 films
(20 mm × 20 mm) and then the films were placed in a purifier hori-
zontal clean bench at ambient temperature (22 ± 2 ◦ C) and relative
humidity (50 ± 5%) under visible light irradiation (20 W daylight
lamp). After given times, the microbial cells were washed from the
films for 5 times with 2 mL sterile 0.85% saline for each time and the
concentrations were determined using the spread plating colony
enumeration technique (Barbosa et al., 1995) with the number of
colonies less than 100. For each assay, the antimicrobial activity
evaluation was conducted in triplicate and the average value was
 X. Zhang et al. / Carbohydrate Polymers 169 (2017) 101–107
presented with the standard deviation for all data. Statistical analy-
sis was conducted by SPSS 17.0 with Duncan’s Multiple Range Test
(P < 0.05).
2.5. Cellular substances’ leakage assays
2.5.1. Enzymes’ analytical assay (Chung & Chen, 2008)
The E. coli cellular suspension (∼106 CFU/mL) was treated with
the chitosan-TiO2 film for given times and 1 mL of supernatant was
obtained by centrifugation (8000 rpm, 5 min) for the measurement
of alkaline phosphatase and glucose-6-phosphate (G-6-P) dehydro-
genase. For alkaline phosphatase, 0.1 mg p-nitrophenylphosphate
and Tris-HCl buffer (0.5 M, pH = 8) were added and the mixture
(total volume: 1 mL) was set at 28 ◦ C to conduct the reaction.
The optical absorbance at 420 nm was monitored by UV–vis spec-
trophotometer and a unit of alkaline phosphatase was defined
as the amount of enzyme catalyzing the production of 1 ␮M p-
nitrophenol-equivalent in 1 min. For G-6-P dehydrogenase, the
supernatant was treated with phenol/chloroform mixture and then
centrifuged at 8000 rpm for 5 min before the analytical assay in
order to prevent the interference from other molecules such as
proteins. 1 mL reaction mixture containing 0.05 M Tris–HCl buffer
(pH = 8), 0.01 M CaCl2 , 1.0 ␮mol glucose-6-phosphate, and 0.4 ␮mol
Triphosphopyridine nucleotide (TPN), was set at 28 ◦ C for the
reduction of TPN. The optical absorbance at 340 nm was moni-
tored and a unit of G-6-P dehydrogenase was defined as the amount
of enzyme catalyzing the reduction of 1 ␮M of TPN-equivalent in
1 min. All experiments were conducted in triplicate and the average
values were presented with their standard deviation.
2.5.2. Nucleotides’ analytical assay (Chung & Chen, 2008)
The supernatant of E. coli cellular suspension (∼106 CFU/mL)
after treatment with the chitosan-TiO2 film for given times was pre-
treated using the similar method as for G-6-P dehydrogenase assay
and the amount of released nucleotides was determined using a
UV–vis spectrophotometer (OD260nm ). All experiments were con-
ducted in triplicate and the average values were presented with
their standard deviation.
2.6. Food preservation evaluation
The application of the prepared chitosan-TiO2 film in food
preservation as antimicrobial packaging material was studied using
red grapes purchased from a local fruit store. Uniform red grape
samples were singly packed and sealed in plastic wrap, pure
chitosan, or chitosan-TiO2 films (50 mm × 50 mm) and three red
grapes were used for each film. All the samples were stored in a
constant-temperature incubator at 37 ◦ C.
3. Results and discussion
3.1. Characterization of chitosan-TiO2 film
3.1.1. Morphologies study by SEM
SEM was employed for studying the surface and cross-section
morphologies of the pure chitosan and chitosan-TiO2 films and the
results are shown in Fig. 1. As shown in Fig. 1(a) and (b), the pure
chitosan film possesses a very smooth surface and compact struc-
ture. After the addition of TiO2 nano-powder, although the surface
of the chitosan-TiO2 film becomes rougher (Fig. 1(c)) because of
the embedded and well-dispersed TiO2 into the chitosan matrix
(Xiao et al., 2014), the structure is as compact as the pure chitosan
film with homogeneous cross-section (Fig. 1(d)). The thickness of
chitosan-TiO2 film is 70 ␮m, which is slightly thicker than that of
the pure chitosan film (66 ␮m) because of the addition of TiO2 nano-
powder. This result demonstrates the excellent phase compatibility
103
Table 1
Tensile strength (TS) and elongation at break (E) of pure chitosan and chitosan-TiO2
films.a
Films TS (MPa) E (%)
Chitosan film 24.43 ± 1.36 15.23 ± 0.78
Chitosan-TiO2 film 46.33 ± 1.88 25.77 ± 2.91
a
Data as average ± standard deviation (p < 0.05).
between chitosan and TiO2 , which agrees with the previous work
of Zhang et al. (2016).
3.1.2. Mechanical properties
The mechanical properties (tensile strength and elongation at
break) of the pure chitosan and chitosan-TiO2 films are shown in
Table 1. The addition of TiO2 nano-powder resulted in a signifi-
cantly increased TS (by 89.64%) and E (by 69.21%) for chitosan-TiO2
film when compared with the pure chitosan film. The increasing TS
resulted from the increased internal friction in the chitosan film
matrix when TiO2 nanoparticles were added to create a crystal
structure (Sadeghi & Shahedi, 2016). It was worth noting that the
TS of the chitosan-TiO2 composite film was much higher than that
of polyethylene (13–28 MPa) (Tan et al., 2015), which was benefi-
cial for its application in food packaging. The increased E resulted
from the fine compatibility of the two components (Zhang et al.,
2016) which contributed to weaken the intermolecular interaction
within the chitosan film matrix (Tan et al., 2015).
3.1.3. Wettability properties
The water contact angle, which is the angle between the sur-
face and the tangent line at the point of water droplet contacting
with the surface, determines the wettability of a surface and is
used as an indicator of the relative hydrophilicity of a surface (Al-
Naamani et al., 2016; Romero-Bastida et al., 2016). A low contact
angle indicates a relative good wetting and hydrophilicity property.
The Water contact angle of the prepared chitosan and chitosan-
TiO2 films were measured and the results are shown in Fig. 2. For
the pure chitosan film, the contact angle of was 98.0 ± 1.2◦ , which
was similar to the previous report from Ferreira, Nunes, Castro,
Ferreira, and Coimbra (2014). For the chitosan-TiO2 film, the con-
tact angle decreased to 44.4 ± 1.5◦ , illustrating the significantly
enhanced hydrophilicity when incorporating TiO2 nano-powder
because of the highly hydrophilic nature of TiO2 (Nayak, Jyothi,
Balakrishna, Padaki, & Ismail, 2015). The enhanced hydrophilic-
ity, although resulting in a relative poor resistance against water
(Shahbazi, Rajabzadeh, & Ahmadi, 2017), was however beneficial
for the contact between the microbial cells and the film which
facilitated the antimicrobial effects.
3.1.4. UV–vis spectroscopy analysis
In order to the evaluate the interactions between the pre-
pared chitosan-TiO2 film and visible light irradiation, the UV–vis
transmittance spectrum of the chitosan-TiO2 film was recorded
as shown in Fig. 3 with the pure chitosan film and TiO2 nano-
powder as references. Compared with the pure chitosan film and
TiO2 nano-powder, the prepared chitosan-TiO2 film possessed a
decreased transmittance in the visible region which contained light
absorption and scattering. The scattering of light occurred because
of the presence of TiO2 nanoparticles and could possibly facilitate
the further interactions between visible light and chitosan-TiO2
composites.
3.2. Antimicrobial activity
The antibacterial activity of the prepared chitosan-TiO2 film
was analyzed against four typical food-borne pathogenic microbes,
104 X. Zhang et al. / Carbohydrate Polymers 169 (2017) 101–107

Fig. 1. (a) SEM surface image of pure chitosan film with inserted schematic diagram; (b) SEM cross-section image of pure chitosan film; (c) SEM surface image of chitosan-TiO2
film with inserted schematic diagram; (b) SEM cross-section image of chitosan-TiO2 film.

Fig. 2. Water contact angles of the pure chitosan film (a) and chitosan-TiO2 film (b).

E. coli (gram-negative bacteria), S. aureus (gram-positive bacteria),

Fig. 3. UV–vis spectra of the pure chitosan film (a), TiO2 nano-powder (b), and
chitosan-TiO2 film (c).
C. albicans (fungi), and A. niger (molds), using the colony counting
method. The results are shown in Fig. 4, demonstrating that the
bactericidal ratios of the tested strains for 4 h were all in excess of
90% and the chitosan-TiO2 film possessed the highest antimicrobial
activity against E. coli with the bactericidal ratio over 99.9% possibly
resulted from the thinner cell wall of gram negative bacteria (Xiao
et al., 2015). Complete sterilization for all the tested strains could
be achieved after 12 h of antimicrobial treatment illustrating the
efficient antimicrobial activity against bacteria, fungi, and molds.
In order to further evaluate the effect of TiO2 nano-powder addi-
tion on the antimicrobial activity of chitosan-TiO2 film, the total
colony forming units of E. coli after contacting with the pure chi-
tosan and chitosan-TiO2 films for different times were recorded as
shown in Fig. 5. In the first 2 h, the concentration of E. coli cells
decreased gradually from 3.5 × 106 CFU/mL to about 105 CFU/mL
for both films. In the following 4 h, the E. coli cells’ concentration
continued to decrease and the final bactericidal ratio approached
100% only for the chitosan-TiO2 film treatment group. For the pure
chitosan film treatment, the E. coli cells’ concentration did not show
an obvious decrease and fluctuated around 105 CFU/mL. The above
 X. Zhang et al. / Carbohydrate Polymers 169 (2017) 101–107
Fig. 4. Bactericidal ratios of the chitosan-TiO2 film against E. coli, S. aureus, C. albi-
cans, and A. niger for different times.
Fig. 5. Antimicrobial kinetics of the pure chitosan and chitosan-TiO2 films towards
E. coli.
results illustrated that pure chitosan possessed limited antimi-
crobial activity because of the presence of surface amino groups
(Zemljič et al., 2013), and was not efficient enough in protect-
ing food from pathogenic microbial contamination. The addition
of TiO2 nano-powder enhanced the antimicrobial activity of the
pure chitosan film through the formation of reactive oxygen species
(ROS) via light mediated charge separation (Abbas et al., 2016). Fur-
thermore, the embedment of TiO2 nano-powder into the chitosan
matrix helped TiO2 to be activated by visible light (Fig. 3) which
facilitate its application in food packaging with efficient antimicro-
bial activity against pathogenic microbes under ambient condition.
3.3. Cellular substances’ leakage
In order to study the effect of chitosan-TiO2 film treatment
on the cell structure, the leakage of cellular substances was ana-
lyzed including alkaline phosphatase (extracellular enzyme), G-6-P
dehydrogenase (in the cell membrane), and nucleotides (in the
cytoplasm) (Chung & Chen, 2008). The results are shown in Fig. 6.
The leakage of enzymes could be observed during the antimi-
crobial treatment of the chitosan-TiO2 film towards E. coli cells
as shown in Fig. 6(a). The leakage of alkaline phosphatase, as an
105
Fig. 6. Effect of chitosan-TiO2 film on the leakage of cellular substances from E. coli:
(a) enzymes; (b) nucleotides.
extracellular enzyme, reached a steady state within 2 h. For G-6-P
dehydrogenase, which is located in the cell membrane, the leak-
age plateau appeared after 5 h. The leakage of enzymes illustrated
that the antimicrobial action of the chitosan-TiO2 film could affect
the structure of the cell membrane and therefore change its per-
meability, resulting in the leakage of intracellular substances such
as nucleotides (Fig. 6(b)). The above results revealed that microor-
ganism cells could be killed through the damage of the membrane
integrity and leakage of cellular substances caused by the antimi-
crobial treatment of the chitosan-TiO2 film.
3.4. Food preservation application
In order to evaluate the potential application of the prepared
chitosan-TiO2 film in food packaging, the preservation of red grape
was studied with plastic wrap as control. Photos were taken after
6 days of storage at 37 ◦ C as shown in Fig. 7. When packed with plas-
tic wrap, the red grapes changed to dark red with obvious mildew
appearance containing several moldy spots marked with yellow
circles and sticky juice leaked to the surface (Fig. 7(a)). Furthermore,
the red grapes packed with plastic wrap emitted a strong rotten
smell. The red grapes packed with the pure chitosan and chitosan-
TiO2 films were still bright red without putridity and the surface
maintained smooth without any leakage of the juice (Fig. 7(b) & (c)),
illustrating the satisfactory preservation effect of the films for food
packaging application. Preservation of red grapes lasted 15 days
106 X. Zhang et al. / Carbohydrate Polymers 169 (2017) 101–107
Fig. 7. Preservation of red grape packed in different materials at 37 ◦ C for 6 days: (a) plastic wrap; (b) pure chitosan film; (c) chitosan-TiO2 film.
with pure chitosan film and 22 days with chitosan-TiO2 film before
mildew occurred. The results showed that the prepared chitosan-
TiO2 film was a promising food packaging material which could
protect food from microbial infection and extend its shelf life.
4. Conclusion
TiO2 nano-powder was successfully introduced into the chi-
tosan matrix with good dispersion and a chitosan-TiO2 composite
film was prepared. The wettability and mechanical properties
of the composite film were enhanced by the addition of TiO2
nano-powder. The chitosan-TiO2 film possessed a decreased
transmittance in the visible light region which facilitated its pho-
tocatalytic antimicrobial effect under ambient condition. Complete
sterilization for all the tested strains in 12 h illustrated the efficient
antimicrobial activity of the chitosan-TiO2 film against bacteria,
fungi, and molds. The antimicrobial effect of the chitosan-TiO2
film damaged the membrane integrity and provoked the leakage
of cellular substances. The prepared chitosan-TiO2 film could suc-
cessfully protect red grapes from microbial infection and extend
their shelf life. The present paper presented a novel composite
film for food packaging applications with satisfactory antimicrobial
activity.
Acknowledgements
The authors express their thanks for the supports from
the National Natural Science Foundation of China (21525625),
the National Basic Research Program (973 Program) of China
(2014CB745100), and the (863) High Technology Project
(2013AA020302) and the Chinese Universities Scientific Fund
(JD1417).
References
Abbas, N., Shao, G. N., Haider, M. S., Imran, S. M., Park, S. S., Jeon, S. J., et al. (2016).
Inexpensive sol-gel synthesis of multiwalled carbon nanotube-TiO2 hybrids for
high performance antibacterial materials. Materials Science and Engineering: C,
68, 780–788.
Al-Naamani, L., Dobretsov, S., & Dutta, J. (2016). Chitosan-zinc oxide nanoparticle
composite coating for active food packaging applications. Innovative Food
Science & Emerging Technologies, 38, 231–237.
Aljawish, A., Muniglia, L., Klouj, A., Jasniewski, J., Scher, J., & Desobry, S. (2016).
Characterization of films based on enzymatically modified chitosan derivatives
with phenol compounds. Food Hydrocolloids, 60, 551–558.
Alkan, D., Aydemir, L. Y., Arcan, I., Yavuzdurmaz, H., Atabay, H. I., Ceylan, C., et al.
(2011). Development of flexible antimicrobial packaging materials against
Campylobacter jejuni by incorporation of gallic acid into zein-based films.
Journal of Agricultural and Food Chemistry, 59(20), 11003–11010.
Atarés, L., & Chiralt, A. (2016). Essential oils as additives in biodegradable films and
coatings for active food packaging. Trends in Food Science & Technology, 48,
51–62.
Barbosa, H. R., Rodrigues, M. F. A., Campos, C. C., Chaves, M. E., Nunes, I., Juliano, Y.,
et al. (1995). Counting of viable cluster-forming and non cluster-forming
bacteria: A comparison between the drop and the spread methods. Journal of
Microbiological Methods, 22(1), 39–50.
Bie, P., Liu, P., Yu, L., Li, X., Chen, L., & Xie, F. (2013). The properties of antimicrobial
films derived from poly (lactic acid)/starch/chitosan blended matrix.
Carbohydrate Polymers, 98(1), 959–966.
Chang, H., Lv, J., Zhang, H., Zhang, B., Wei, W., & Qiao, Y. (2017). Photoresponsive
colorimetric immunoassay based on chitosan modified AgI/TiO2
heterojunction for highly sensitive chloramphenicol detection. Biosensors and
Bioelectronics, 87, 579–586.
Chen, G. G., Qi, X. M., Guan, Y., Peng, F., Yao, C. L., & Sun, R. C. (2016). High strength
hemicellulose-based nanocomposite film for food packaging applications. ACS
Sustainable Chemistry & Engineering, 4(4), 1985–1993.
Chung, Y. C., & Chen, C. Y. (2008). Antibacterial characteristics and activity of
acid-soluble chitosan. Bioresource Technology, 99(8), 2806–2814.
Fan, X., Chen, K., He, X., Li, N., Huang, J., Tang, K., et al. (2016).
Nano-TiO2 /collagen-chitosan porous scaffold for wound repairing.
International Journal of Biological Macromolecules, 91, 15–22.
Ferreira, A. S., Nunes, C., Castro, A., Ferreira, P., & Coimbra, M. A. (2014). Influence of
grape pomace extract incorporation on chitosan films properties. Carbohydrate
Polymers, 113, 490–499.
Hu, Z., Hong, P., Liao, M., Kong, S., Huang, N., Ou, C., et al. (2016). Preparation and
characterization of chitosan-agarose composite films. Materials, 9(10), 816.
Kamal, T., Anwar, Y., Khan, S. B., Chani, M. T. S., & Asiri, A. M. (2016). Dye adsorption
and bactericidal properties of TiO2 /Chitosan coating layer. Carbohydrate
Polymers, 148, 153–160.
Kamari, Y., & Ghiaci, M. (2016). Preparation and characterization of
ibuprofen/modified chitosan/TiO2 hybrid composite as a controlled
drug-delivery system. Microporous and Mesoporous Materials, 234, 361–369.
Kuo, C. S., Tseng, Y. H., Huang, C. H., & Li, Y. Y. (2007). Carbon-containing
nano-titania prepared by chemical vapor deposition and its
visible-light-responsive photocatalytic activity. Journal of Molecular Catalysis A:
Chemical, 270(1), 93–100.
Leceta, I., Etxabide, A., Cabezudo, S., de La Caba, K., & Guerrero, P. (2014). Bio-based
films prepared with by-products and wastes: Environmental assessment.
Journal of Cleaner Production, 64, 218–227.
Lee, S. Y., & Park, S. J. (2013). TiO2 photocatalyst for water treatment applications.
Journal of Industrial and Engineering Chemistry, 19(6), 1761–1769.
Ma, J., Wei, Y., Liu, W. X., & Cao, W. B. (2009). Preparation of nanocrystalline
Fe-doped TiO2 powders as a visible-light-responsive photocatalyst. Research on
Chemical Intermediates, 35(3), 329–336.
Ma, W., Li, J., Liu, Y., Ren, X., Gu, Z. G., Xie, Z., et al. (2016). Preparation and
characterization of excellent antibacterial TiO2 /N-halamines nanoparticles.
Colloids and Surfaces A: Physicochemical and Engineering Aspects, 506, 284–290.
Mallakpour, S., & Madani, M. (2015). Effect of functionalized TiO2 on mechanical,
thermal and swelling properties of chitosan-based nanocomposite films.
Polymer-Plastics Technology and Engineering, 54(10), 1035–1042.
Matsunaga, T., Tomoda, R., Nakajima, T., & Wake, H. (1985). Photoelectrochemical
sterilization of microbial cells by semiconductor powders. FEMS Microbiology
Letters, 29(1-2), 211–214.
Nayak, V., Jyothi, M. S., Balakrishna, R. G., Padaki, M., & Ismail, A. F. (2015).
Preparation and characterization of chitosan thin films on mixed-matrix
membranes for complete removal of chromium. ChemistryOpen, 4(3), 278–287.
Qian, T., Su, H., & Tan, T. (2011). The bactericidal and mildew-proof activity of a
TiO2 -chitosan composite. Journal of Photochemistry and Photobiology A:
Chemistry, 218(1), 130–136.
Romero-Bastida, C. A., Tapia-Blácido, D. R., Méndez-Montealvo, G., Bello-Pérez, L.
A., Velázquez, G., & Alvarez-Ramirez, J. (2016). Effect of amylose content and
nanoclay incorporation order in physicochemical properties of
starch/montmorillonite composites. Carbohydrate Polymers, 152, 351–360.
Sadeghi, K., & Shahedi, M. (2016). Physical, mechanical, and antimicrobial
properties of ethylene vinyl alcohol copolymer/chitosan/nano-ZnO (ECNZn)
nanocomposite films incorporating glycerol plasticizer. Journal of Food
Measurement and Characterization, 10(1), 137–147.
Saha, N. R., Sarkar, G., Roy, I., Rana, D., Bhattacharyya, A., Adhikari, A., et al. (2016).
Studies on methylcellulose/pectin/montmorillonite nanocomposite films and
their application possibilities. Carbohydrate Polymers, 136, 1218–1227.
Shahbazi, M., Rajabzadeh, G., & Ahmadi, S. J. (2017). Characterization of
nanocomposite film based on chitosan intercalated in clay platelets by electron
beam irradiation. Carbohydrate Polymers, 157, 226–235.
 X. Zhang et al. / Carbohydrate Polymers 169 (2017) 101–107
Song, N. B., Lee, J. H., Al Mijan, M., & Song, K. B. (2014). Development of a chicken
feather protein film containing clove oil and its application in smoked salmon
packaging. LWT-Food Science and Technology, 57(2), 453–460.
Tajik, S., Maghsoudlou, Y., Khodaiyan, F., Jafari, S. M., Ghasemlou, M., & Aalami, M.
(2013). Soluble soybean polysaccharide: A new carbohydrate to make a
biodegradable film for sustainable green packaging. Carbohydrate Polymers,
97(2), 817–824.
Tan, Y. M., Lim, S. H., Tay, B. Y., Lee, M. W., & Thian, E. S. (2015). Functional
chitosan-based grapefruit seed extract composite films for applications in food
packaging technology. Materials Research Bulletin, 69, 142–146.
Verbruggen, S. W., Keulemans, M., Filippousi, M., Flahaut, D., Van Tendeloo, G.,
Lacombe, S., et al. (2014). Plasmonic gold-silver alloy on TiO2 photocatalysts
with tunable visible light activity. Applied Catalysis B: Environmental, 156,
116–121.
Wang, Y., Wu, Y., Yang, H., Xue, X., & Liu, Z. (2016). Doping TiO2 with boron or/and
cerium elements: Effects on photocatalytic antimicrobial activity. Vacuum, 131,
58–64.
Xiao, G., Zhang, X., Zhao, Y., Su, H., & Tan, T. (2014). The behavior of active
bactericidal and antifungal coating under visible light irradiation. Applied
Surface Science, 292, 756–763.
Xiao, G., Zhang, X., Zhang, W., Zhang, S., Su, H., & Tan, T. (2015).
Visible-light-mediated synergistic photocatalytic antimicrobial effects and
mechanism of Ag-nanoparticles@chitosan-TiO2 organic-inorganic composites
for water disinfection. Applied Catalysis B: Environmental, 170, 255–262.
107
Xu, W., Xie, W., Huang, X., Chen, X., Huang, N., Wang, X., et al. (2017). The graphene
oxide and chitosan biopolymer loads TiO2 for antibacterial and preservative
research. Food Chemistry, 221, 267–277.
Yang, M., Xia, Y., Wang, Y., Zhao, X., Xue, Z., Quan, F., et al. (2016). Preparation and
property investigation of crosslinked alginate/silicon dioxide nanocomposite
films. Journal of Applied Polymer Science, 133(22), 43489–43497.
Yun, Y. H., Yun, J. W., Yoon, S. D., & Byun, H. S. (2016). Physical properties and
photocatalytic activity of chitosan-based nanocomposites added titanium
oxide nanoparticles. Macromolecular Research, 24(1), 51–59.
Zemljič, L. F., Tkavc, T., Vesel, A., & Šauperl, O. (2013). Chitosan coatings onto
polyethylene terephthalate for the development of potential active packaging
material. Applied Surface Science, 265, 697–703.
Zhang, J., Liu, Y., Li, Q., Zhang, X., & Shang, J. K. (2013). Antifungal activity and
mechanism of palladium-modified nitrogen-doped titanium oxide
photocatalyst on agricultural pathogenic fungi Fusarium graminearum. ACS
Applied Materials & Interfaces, 5(21), 10953–10959.
Zhang, W., Chen, J., Chen, Y., Xia, W., Xiong, Y. L., & Wang, H. (2016). Enhanced
physicochemical properties of chitosan/whey protein isolate composite film
by sodium laurate-modified TiO2 nanoparticles. Carbohydrate Polymers, 138,
59–65.