Accepted Manuscript

Using photo-modification to compatibilize nano-ZnO in

development of starch-kefiran-ZnO green nanocomposite as food
packaging material

Iman Shahabi-Ghahfarrokhi, Amin Babaei-Ghazvini

PII: S0141-8130(18)35592-2
DOI: https://doi.org/10.1016/j.ijbiomac.2018.11.241
Reference: BIOMAC 11118
To appear in: International Journal of Biological Macromolecules
Received date: 17 October 2018
Revised date: 23 November 2018
Accepted date: 26 November 2018

Please cite this article as: Iman Shahabi-Ghahfarrokhi, Amin Babaei-Ghazvini , Using
photo-modification to compatibilize nano-ZnO in development of starch-kefiran-ZnO
green nanocomposite as food packaging material. Biomac (2018), https://doi.org/10.1016/
j.ijbiomac.2018.11.241

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 ACCEPTED MANUSCRIPT

Using photo-modification to compatibilize nano-ZnO in development of

starch-kefiran-ZnO green nanocomposite as food packaging material

Iman Shahabi-Ghahfarrokhia*, Amin Babaei-Ghazvinib

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a) Department of Food Science and Technology, Faculty of Agriculture, University

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of Zanjan, 45371-38791, Zanjan, Iran

b) School of Packaging, Michigan State University, East Lansing, MI, USA

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*Corresponding author: i.shahabi@znu.ac.ir, Tel.: +98 24 3305 2339, Fax: +98 24
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3305 2348
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Abstract

In this study, the effect of UV-C light on starch-kefiran-ZnO (1%) primary solution in different

exposure times (1, 6, and 12 h) was investigated. Starch-kefiran-ZnO (SKZ) solution was

modified by UV irradiation in different time periods. Also, nano-ZnO (ZN) was used as a photo-

initiator and reinforcement agent, simultaneously. Mechanical properties of the films were

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affected after the UV treatment. The tensile strength increased because of the enhanced

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interaction between the biopolymer mixture and nano filler but elongation at break was

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decreased. WVP decreased about 16% and dwindled to 2.08×10−10 g m−1 s−1 Pa−1. Water related

properties (i.e. moisture content, moisture absorption, and solubility in water) of the films

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decreased by UV-C exposure. On the other hand, UV absorption and water contact angle
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increased because of the better distribution of the ZNs in polymer matrix after the UV exposure.

Better compatibility of the ZNs and the biopolymer matrix after UV treatment was confirmed by
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the SEM micrographs. Comparison of FTIR spectra before and after UV exposure showed slight
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shifts. It was due to some formed or deformed bonds inside of the nanocomposite matrix. The

modified SKZ by UV could be an appropriate process to sanitizing and food packaging

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concurrently. As well as UV can be used as a nano-ZnO compatibilizer in food packaging

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materials.
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Keyword: Starch, Kefiran, Zinc oxide, UV ray, Nanocomposite, Food packaging

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1 Introduction

In recent years, the environmental pollution resulting from petroleum-based packaging materials

has become a serious problem [1]. The use of biopolymers for sustainable development and

environmental preservation has become universal in the last 20 years, due to their several

advantages including non-toxicity, biodegradability, wide availability and biocompatibility,

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especially when compared to their synthetic counterparts [2].

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The use of nanomaterials as fillers in nanocomposites have many advantages along with the

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reinforcement of several physical and mechanical properties [3]. Food packaging materials with

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proper mechanical strength, barrier properties against water, water vapor, and other gases,

thermal stability, recyclability, and biodegradability as well as functional properties such as

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antimicrobial and antioxidant activities are highly desirable for food safety and extending the
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shelf-life of packaged foods[4]. Currently, the materials used in packaging industries are

dominated by petroleum based plastic materials produced from fossil fuels since they are
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relatively cheap and convenient to use with excellent process-ability and durability[5].
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Over the past decades, in the context of sustainable material development, attentions in
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biopolymer based nanocomposites are increased considerably owing to their biodegradability

and biocompatibility for potential application in biotechnology and food packaging material.
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Carbohydrate based polymers i.e. starch[6], pullulan[7, 8], and kefiran[9, 10]; protein based

polymers i.e. whey protein [7, 8], soy protein [11], have been investigated to alternate

conventional non-degradable polymers.

Among the biopolymers, starch has been considered as one of the most promising candidates for

this regard because of cheapness, world-wide availability, abundant, renewability and

biodegradability[6].

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Kefiran is a water-soluble exopolysaccharide which produce by kefir microflora and extract

from kefir grains [12]. It contains approximately equal amounts of glucose and galactose[10].

Kefiran was proposed as a packaging material from 2009 and several studies were published

about its modification and characterization [10, 13, 14]. Kefiran is a unique biopolymer with

comparable packaging properties with some of petroleum based polymers[13]. As well as several

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researches were showed its functional properties i.e. antibacterial properties [15], antifungal and

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antitumor activity [16, 17] and modulating the gut immune system [18]. Kefiran is GRAS

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(generally recognized as safe)[19]biopolymer which can form gels alone at low temperature and

edible films with good mechanical and barrier properties [20].

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The application of the bio-polymers such as starch and kefiran films are limited by mechanical
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properties and barrier-ability. In addition, retrogradation of the starch chains leads to an

undesired change in thermo mechanical properties [21]. Various modification methods, including
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cross linking by using ionizing rays e.g. UV-irradiation[22], γ-irradiation [12] chemical
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compounds [23], development of bionanocomposites [6, 24], combination of two biopolymers

[8, 14] has also been used by several authors to improve the physical properties and limitations
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of bio-polymers. Among these methods, ultraviolet (UV) radiation has been interested in by
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several researchers due to its low cost, availability, and environmental-friendly [22, 24-26].
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According to ISO classification solar irradiance standard, the UV irradiation is divided into

UV-A (315-400 nm), UV-B (280-315 nm) and UV-C (200-280 nm) [27]. UV-ray absorb by

double bonds and aromatic rings, creating free radical formation in amino acids and can lead to

the formation of intermolecular covalent bond [28].

Nano-ZnO (ZN) is found to possess high photocatalytic efficiency. It is more biocompatible

than TiO2 and it can highly absorb UV light [29]. ZN is low cost nano filler which exhibits

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extensive promising applications [30]. ZN shows unique catalytic, electrical (i.e. piezo and pyro

electric) and optical properties [31], photostability [32], biocompatibility [33] and

biodegradability [34]. As well as ZN was interested as antimicrobial agent in active packaging

[35]. ZN in aqueous solution under UV radiation have photo-radical feature that can produce

reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) and superoxide ions [29].

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Upon irradiation, valence band electrons (e-) are promoted to the conduction band and leaving a

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hole (h+) behind (Eq. (1)).The holes at the ZN valence band can oxidize adsorbed water or

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hydroxide ions to produce hydroxyl radicals (Eq. (2)). Electron in the conduction band on the

catalyst surface can reduce molecular oxygen to superoxide anion (Eq. (3)). This radical may

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form organic peroxides or hydrogen peroxide in the presence of organic scavengers (Eq. (4)).
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The hydroxyl radical is a powerful oxidizing agent and attacks to organic compounds[36].
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ZnO + hν→ e− + h+ (1)

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h+ + H2O → •OH + H+ (2)

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e− + O2→ O2•− (3)

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O2•− + HO2• + H+ → H2O2 + O2 (4)

Sun et al [37]reported UV irradiation will generate electron-hole pairs in the ZN surface, and

some of the holes can react with lattice oxygen to form surface oxygen vacancies. Meanwhile,

water and oxygen may compete to dissociatively adsorb on them. The defective sites are

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kinetically more favorable for hydroxyl adsorption than oxygen adsorption. As a result, the

surface hydrophilicity is increase under UV radiation. Dark storage for several days causes the

surface character reconverted from hydrophilic to hydrophobic[37].

To avoid the aggregation of ZNs some compatibilizers (i.e. tetra-ethylene glycol and bovine

serum albumin) were used in previous studies[38]. Nevertheless, it seems that ZN particle size,

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pH of the filmogenic solution, film preparation method, and the biopolymer type were the key

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factors in the aggregation of ZN in nanocomposites[10, 24]

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The main purpose of current study is investigating the effect of UV-C irradiation on

filmogenic solution of Starch-Kefiran-ZnO (SKZ). According to the same studies and our

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previous experiments seem UV radiation could improve the compatibility between biopolymers
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and ZN. Furthermore, the formation of cross-links is probable. Thus, our research team

appointed this level of concentration in this study to modify the functional properties of the thin
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film as a food packaging material.

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2 Materials and methods

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2.1 Material

Wheat starch (11-13 % moisture) was obtained from Khosheh-Zarin starch industry (Bojnord,
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Iran). ZNs provided by Tecnan (Tecnología Navarra de Nanoproductos S.L, Spain) were used as
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they arrived. According to the producer’s catalogue, the average size and specific surface area of

ZNs were 20–30 nm and 35–50 m2/g, respectively. The analytical grade chemicals including

magnesium nitrite and glycerol were purchased from Merck Co., (Darmstadt, Germany). Sodium

chloride and calcium chloride were also provided from Dr. Mojalali Co., (Tehran, Iran).

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2.1.1 Starter culture

Kefir grains utilized as a starter culture in this work, were purchased from a domestic supplier

in Tehran, Iran. The grains were grown in skimmed milk (1:5, w/w) at 25°C. Skimmed milk was

renewed daily to keep the kefir grains’ alive.

2.1.2 Isolation and purification of kefiran

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Kefiran, exo-polysaccharide, was extracted from the kefir grains by the described method by

Piermaria et al [39]. In brief, a weighed amount of kefir grains was added to boiling water (1:10)

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and continuously stirred for an hour. After reaching to the uniformity, the mixture was

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centrifuged at 10000 ×g for 15 mins. An equal volume of chilled ethanol was then added to

precipitate the polysaccharides secreted in the supernatant and the mixture was kept at -18°C
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overnight. After that time, the pellets were collected by centrifuging at 10000 ×g at 4°C for 20
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min. The precipitates were re-dissolved in hot distilled water and the precipitation procedure was

repeated twice. The total polysaccharide of kefiran was 92% (as glucose equivalent) according to
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the phenol sulfuric method [40].

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2.2 Preparation of the film solution

An aqueous solution of starch (5%) was prepared and heated until its gelatinization (80°C for
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60 min). An aqueous solution of 2 wt% kefiran was prepared. Starch/kefiran (SK) film solution
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was also prepared by mixing starch solution and kefiran solution (1:1), but the dry base ratio of

the starch-kefiran in final films were (5:2) respectively. Glycerol as plasticizer (40 wt% of dry

base) was also added. Then the solution was stirred for 15 min. The slurry of ZN was prepared to

achieve final concentration in filmogenic solution (1wt% (dry base)). After that stirred for 15

min and then sonicated by an ultrasonic homogenizer (Dr. Hielscher, Teltow, Germany) at

amplitude 50%, cycle 1, and 50 Hz for 30 min. The slurry of ZN were added to the SK solution

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and mixed for 10 min. 50 ml of the filmogenic solution were cast onto flat leveled, non-stick

polystyrene microbial plates (15 cm diameter) and were dried at room temperature (25 °C) as

control sample. The SK-ZN nanocomposite film is named hereafter SKZ. The dried films were

peeled off from the casting surface after 48 hours.

2.3 UV irradiation

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SKZ solution placed under three UV-C lamps to coverage all over the solution surface. The

UV-C lamps (TUV 8W G5 FAM, Phillips, Holland) with maximum emission at 254 nm were in

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parallel at a distance of 5 cm from surface of the solution (Figure 1). SKZ solutions were stirred

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at 50 rpm and 30 ˚C by a magnetic stirrer. The solution were stirred during this time with using a

magnetic stirrer. Film forming solution were exposed to1 hour UV-C (SKZU1), 6 hour UV-C
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(SKZU6) and 12 hour UV-C (SKZU12). 50 ml of the irradiated solutions were cast onto flat
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leveled, non-stick disposable microbial plates and were dried at room temperature. The dried

films were peeled off from the casting surface and storage in a dark situation inside of an
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aluminum foil (dark storage). This process reconverted the highly hydrophilic films character to
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their original states[37]. Before any analysis, all film specimens were conditioned inside

desiccators containing saturated magnesium nitrite solution to ensure a relative humidity of 50-
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55 % at 25 °C for at least 48h.

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Figure 1.

2.4 Microstructural characterization

The morphology of the surface area and cross sections of the specimen films were observed by

field emission scanning electron microscope (FE-SEM, Hitachi, S4160; Japan) with the

accelerating beam at a voltage of 10 kV. The cross sections were prepared by breaking the films

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samples in liquid nitrogen and mounting on aluminum stubs using a double-sided tape. Then, the

film specimens were sputtered with gold using a sputter coater.

2.5 Intrinsic viscosity determination

The intrinsic viscosity of the SK and SKZ solutions were determined by double extrapolation

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to zero concentration of the Huggins (Eq. (5)) and Kraemer (Eq. (6)) equations, respectively [41]

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ηsp

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= [η] + K H [η]2 C (5)
C

ln(ηrel )

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= [η] + K K [η]2 C (6)
C

Where [η] , ηsp and ηrel is intrinsic, specific and relative viscosities, respectively. ηrel and
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ηsp were estimated with Eqs. (7) and (8), respectively. KH and KK are the Huggins and Kraemer
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coefficients and C is the polymer concentration (w/v). The intrinsic viscosity was calculated
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using an Ubbelohde type of capillary viscometer at 25 °C.

t slu
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ηrel = (7)
t slv
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ηsp = ηrel -1 (8)

Where tslu and tslv are average efflux time polymer solution in water and water, respectively.
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2.6 Contact angle

Sessile drop method was used for contact angle measurements. A 5 µl droplet of water was

placed on the film surface. The image of the drop was captured using a digital camera (AM2111,

Dino-Light, Taiwan). The contact angle is defined as the angle between the baseline of the drop

and the tangent line at the point of contact of the water droplet with the surface[42]. Image J

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software (version Java 1.6.0_05) was used to estimate the contact angle. Six drops of water were

used on each film to calculate the contact angle.

2.7 Fourier transform infrared (FTIR) spectroscopy

FTIR spectrum of the film specimens were acquired at room temperature by FTIR

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spectroscopy (IRPrestige-21, Shimadzu, Japan). The specimen solutions were ground with KBr

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powder and pressed into pellets and dried. FTIR spectrums were measured in the wavenumber

range of 4000–400 cm-1 at a resolution of 4 cm−1.

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2.8 Physical characteristics

2.8.1 Film thickness

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The thickness of specimens was measured by a hand-held micrometer with an accuracy of
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0.01 mm. The thickness of ten randomly selected points was measured for each testing specimen
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and the mean value was reported for all experiments.

2.8.2 Moisture content

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The moisture content (MC) of specimens was determined by measuring the weight loss of
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films before and after drying in a laboratory oven (Pars Azma Co., Tehran, Iran) at 105±1 °C

until a constant weight was reached. MC was calculated with Eq. (9) in triplicate.
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m1 - m2
MC = ×100 (9)
m1

Where m1 (g) was the initial weight of the specimens and m2 (g) was the weight of specimen

after oven drying

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2.8.3 Moisture absorption

The moisture absorption (MA) was determined (in three replicates) according to the difference

in the weight of the films before and after conditioning at relative humidity of 50-55 %. The test

specimens were prepared with the dimensions of 20 mm × 20 mm. They were first conditioned at

0% RH (prepared by dried calcium chloride) to reach a constant weight. After weighing (m3 (g)),

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they were conditioned in a desiccator containing magnesium nitrite saturated solution at 25 °C to

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ensure a RH of 50-55%. The sample was weighed at determined intervals until the equilibrium

state was reached (m4 (g)). The moisture absorption of the specimen was calculated with Eq. (10)

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m 4 - m3
MA = 100 (10)
m3
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2.8.4 Film solubility in water
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Solubility in water (SW) was explained as the percentage of dry matter of film solubilized
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after a six-hour immersion in distilled water. Film specimens (20 mm × 20 mm) were dried at

105 ± 1 °C to achieve constant weight (m5 (g)). The dried films were immersed in 50 ml of
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distilled water while stirring for six hours at 25 °C. After that, the pieces of remained film were
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removed and dehydrated at 105 ± 1 ˚C to reach a steady weight (m6 (g)). Three replications were

performed for this test. The SW of the specimens was calculated using Eq. (11).
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m5 - m 6
SW = ×100 (11)
m5

2.9 Colorimetric measurements

Film color was determined using a colorimeter (RGB-1002, Lutron Electronic Enterprise Co.,

Taiwan). Film specimens were placed on a white standard plate (L*= 100, a*=-1, b*=2) and the

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lightness (L) and chromaticity parameters a (red-green) and b (yellow-blue) were measured. All

colors can be described by L values ranging from 0 (black) to 100 (white); negative values of a

(greenness) to positive values (redness); and negative values of b (blueness) to positive values

(yellowness). All measurements were performed in four replicates. The total color difference

(ΔE), whiteness index (WI) and yellowness index (YI) were calculated using Eqs. (12, 13, and

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14), respectively[10].

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ΔE = ( L* - L)2 + (a* - a)2 + (b* - b)2 (12)

WI = 100 - (100 - L) 2 + a 2 + b2 (13)

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142.86  b
YI = (14)
L
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2.9 UV-Vis Spectroscopy
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The UV–visible spectra of the SKZ and SKZU nanocomposites were measured by using a UV-
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2600 spectrophotometer (SPECORD 250 UV/ VIS, Germany). The film samples were placed

directly in the spectrophotometer test cell and the air was used as reference. The optical
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absorbance of the films were measured in the wavelength range of 200–800 nm.
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2.10 Water vapor permeability

ASTM E96 standard was used for measuring the water vapor permeability (WVP) of the

specimens [43]. Glass vials with the diameter of 13 mm and the height of 45 mm, containing

calcium chloride desiccant (0% RH), were used to measure WVP of the films. The vials were

covered with film specimens and the vial mouths were sealed using double side tape.

Furthermore, three similar vials without any desiccant were used to tare the absorption of

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moisture by every film specimen. Each vial was positioned in a desiccator preserved at 75% RH

with a saturated solution of sodium chloride. The difference in RH causes to a driving force of

1753.55 Pa, expressed as water vapor partial pressure at 20 °C. After that the films were fixed,

the weight gain of the whole container and sample was periodically observed and recorded (with

an accuracy of 0.0001 g) every 1 h for 24 h. The slope (S) of the weight vs. time plot (s) (R2≥

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0.96) was divided by the effective film area (A) to obtain the water vapor transmission rate

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(WVTR) Eq. (15). This was multiplied by the thickness of the film and divided by the pressure

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difference between the inside and outside surfaces to acquire the WVP Eq. (16). Three replicates

of each film treatment were tested for WVP test [43, 44].

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S
WVTR = (15)
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A
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WVTR ×X
WVP = (16)
ΔP
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Where A is the effective film area (m2), X is the average film thickness (m) and ΔP is the
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driving force (1753.55 Pa).

2.11 Mechanical properties

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Tensile strength (TS), elongation at break (EB), tensile energy to break (TEB), and Young’s

modulus (YM) were evaluated by mechanical evaluation device (STM-250, Santam Co., Iran)

according to ASTM standard method D882-02 [45]. Films were cut in rectangular strips of 100

mm length and 10 mm wide. All film strips were conditioned at 55% RH for at least 48 h in a

desiccator using saturated magnesium nitrate solution. The films were mounted with an initial

grip separation of 50 mm and stretched at a cross-head speed of 10 mm/min. Three replicates

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were run for each film specimen. TS, EB and TEB were calculated by Eqs. (17, 18, and 19),

respectively.

FMax
TS = (17)
A Min

LMax

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EB = ×100 (18)
L0

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TEB = AStress-Strain (19)

Where FMax is maximum load, AMin is minimum cross-section area, LMax is extension at the

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moment of rupture, L 0 is initial length of specimen, AStress-Strain is area under stress–strain curve.
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YM was calculated by drawing a tangent to the initial linear portion of the force extension
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curve, selecting any point on this tangent and dividing the tensile stress by the corresponding
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strain, Eq (20).

stress
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YM = (20)
strain
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2.12 Statistical analysis

The results obtained from all experiments were analyzed on a completely randomized design
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with the analysis of variance (ANOVA) procedure using SPSS software (Version 16; SPSS Inc.,

USA). Duncan’s multiple range tests were used to compare the difference between mean values

of film specimens’ properties at the 0.05 level of significance.

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3 Results and discussion

3.1 Microstructure

Figure 2 shows the surface and cross-section of the SKZ (1%) nanocomposites before and after

exposure to UV irradiation. SEM images of this nanocomposites show the aggregation of ZNs as

shiny spots on the surface of non-irradiated specimen (S1) in (Figure 2). The UV treated films

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showed homogenous structure on the surface after exposure to UV ray. It is noteworthy that,

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after UV irradiation the distribution of the ZNs in biopolymer matrix came better. But there was

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no significant difference between UV irradiated SKZ at different exposure time. It seems, it was

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due to the change in hydrophobic character to hydrophilic on the surface of the ZNs under UV

radiation in filmogenic solution [37]. In other words, the size of aggregated nano structures
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decreased under exposure to UV. The aggregated ZNs are pinpointed in the surface images by

red pointers. This property of the ZNs has caused the nanostructures dispersed well in the initial
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solution and ultimately this phenomenon prevent to aggregate the ZNs in the film specimens.
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Better dispersion of the nanoparticles in the polymer matrix leads to a powerful interaction
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between ZNs and SK biocomposite. Whereas, other researchers had complained about the lack

of compatibility of ZN with biopolymers[10, 24]. It seems, UV irradiation is a green

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compatibilizing process to develop ZN based nanocomposites.

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Figure 2.

3.2 FTIR

The broad spectrum from 3100 cm−1 to 3600 cm−1 contains the fundamental stretching modes of

hydroxyl groups (OH) due to water and carbohydrates. Spectrum from 3000 to 2800 cm−1

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attributed to the symmetric and anti-symmetric stretching modes of CH in methyl (CH3) and

methylene (CH2) functional group, 1720–1740 cm−1 assigned to C-O stretching, 1700–1580 cm−1

assigned to bending mode of O-H, 1200–900 cm−1 a broad and intense band which assigned to

stretching modes of carbohydrate rings and side group (C -O - C, C- OH, C-H) and 900–700

cm−1 assigned to bending of C-H [46].The small peaks around 500 cm-1 clearly indicates Zn–O

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stretching[47]. Slight shifts in some of the bonds were also observed from the spectra between

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treatments. This might be possibly due to the crosslinking that might have occurred in starch and

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kefiran chains after exposure to UV radiation.

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However, further investigations are required to characterize the same and look for the possible

quantitative changes taking place in the functional groups. As UV light provides a lower energy
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level than gamma irradiation, it is less likely for direct split of C-C or C-H bond of starch or
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kefiran molecules to occur formation of free radicals. Hence, presence of a photosensitizer (i.e.

ZN) to absorb a low-energy photon (i.e. UV ray) and free radicals formation is necessary [48]
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and also leads to macro radical combination by hydrogen abstraction[49].

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As shown in (Figure 3), by increasing UV exposure time, the bond observed at 3426 cm−1 in

SKZ spectrum (OH group) was shifted to lower frequencies (3412, 3398, and 3385 cm−1 in
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SKZU1, SKZU6, and SKZU12, respectively). These results were attributed to the formation of
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hydrogen bonds between hydroxyl groups in SK, and the abundant hydroxyl groups that are

present in the surface of ZNs. The free radicals predispose the matrix of the bionanocomposites

to form cross-links [50]. These certain interactions have been previously observed[51].

Figure 3.

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3.3 Contact angle

Contact angle measurements can be a good way to determine hydrophilic and hydrophobic

character of the surfaces. The contact angle images for specimens are shown in Figure 2-A. The

contact angle of SKZU was higher than non-irradiated SKZ film. It is well-known that the water

contact angle will increase with increasing surface hydrophobicity[52].The results in this study

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showed that by increasing the UV exposure time the water contact angle of specimens was

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increased significantly from 95.63 º to 107.37 º. It seems, it was due to good combination

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between the ZNs and the biopolymer matrix (Figure 2). On the other words, UV irradiation

changes the character of ZN from hydrophobic to hydrophilic[47] and this phenomenon causes a

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good dispersion of the nanoparticles in the liquid state. After that, in absence of UV irradiation,
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the ZNs surface character changed from hydrophilic to hydrophobic slightly [37] and the ZNs on

the surface causes the distortion of the contact line between the solid-liquid interfaces[10]. It
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confirms, the hydrophilic groups were decreased on the surface of the bionanocomposite by UV
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irradiation [6, 24].

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3.4 Intrinsic viscosity

The intrinsic viscosity provides an insight of the hydrodynamic volume occupied by a given
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polymer and the chain length of polymer [39]. According to the Table 3, the intrinsic viscosity of

specimens significantly decreased by increasing UV-C irradiation time. This results was similar

to the results of Bertolini et al [53] and Bhat et al [48]. These researchers have reported the

reduction of intrinsic viscosity of corn and cassava starch[53]and fish gelatin granules[48]under

UV radiation. Although UV irradiation provides lower energy than gamma and electron beam. It

is possible that, with sufficient exposure time or photosensitizer (i.e. ZN), form adequate free

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radicals to induce molecular changes and fragmentation[48]. Starch and kefiran chains were

broken and produced shorter chain polymers, disaccharides and monosaccharides. It is in

agreement with previous studies about the effect of γ-irradiation on kefiran polysaccharide [12]

and UV irradiation on starch polysaccharide [54].

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3.5 Thickness, moisture content, moisture absorption, and solubility in water

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A great drawback to the use of biopolymer materials is moisture sensitivity; thus, any increase in

water resistance of biopolymers was the main purpose of several studies [6, 22, 24]. Table 1

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shows the thickness, MC, MA, SW and contact angle of film specimens. The thickness of SK

films is about 0.12 mm. The thickness of film specimens was not significantly changed with
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increasing UV exposure time. Previous studies have been shown decreasing in the thickness of
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the films[12]. It seems, it depends on polymer type, UV type, UV exposure time, nanoparticle
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type, nanoparticle content, and film preparation method.

Table 1.
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UV radiation changes the character of ZN from hydrophobic to hydrophilic[47]. It means that,

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under the UV light, ZNs surface tend to absorb available OH groups in solution that increase the
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hydrophilicity of ZNs. After turning off the UV light these hydroxyl groups will disappear

slowly[29]. This phenomenon prevent the aggregation of the nanoparticles. Consequently, the

dispersion of the ZNs in the matrix of biopolymers was improved. Better dispersion of the ZNs

could lead to improve moisture sensibility [24].

As the results shows, SW, MA and MC have decreased after UV treatments. But MC and SW

changes were not significant at short UV exposure time (1 h). As described in FTIR results, some

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cross-links were formed during the film formation. It seems, the probable formed cross-links

were effective in decreasing moisture sensibility in the films. As well as, the better distribution

of ZNs by UV radiation (Figure 2) contributed this characters. As we described in contact angle

section, after UV elimination ZNs have hydrophobic surface which could improve the water

related properties of the films. The results is compatible with previous study[12].

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3.6 Mechanical properties

Table 2 shows the effect of UV irradiation period on the mechanical properties (i.e. TS, EB, YM,

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and TEB) of SKZ nanocomposites. The changes in the mechanical properties of the SKZ films

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could be resulted under the influence of two major factors. Firstly, homogenous distribution of

ZNs along with the appropriate interfacial interactions between the biopolymer matrix (Figure
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2), are the key factors to achieve desirable properties in nanocomposites [55]. Secondly,
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increasing the tensile strength could due to the high interaction that occurs between the matrix of

starch-kefiran and ZNs surface. In this way, the density of cross-links was increased by a free
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radical content mechanism [50]. Free radicals e.g. ROS, were formed by exciting ZNs under UV
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irradiation. The free radicals could be reacted with starch-kefiran molecules as cross-linking

agents which increased TS and YM of UV irradiated specimens. On the other hand, the
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limitation of the bionanocomposite chains movement by the formed cross-links decreased EB of

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the samples. It is in agreement with FTIR results (Figure 3). But, it seems that the better

distribution of ZNs is more effective on mechanical properties. This is in agreement with the

previous study [24].

Table 2.

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3.7 Water vapor permeability

Water vapor permeability results can be useful to understand mass transfer mechanisms and

solute-polymer interactions in biodegradable films. In packaging applications, film should be

used to avoid or at least decrease moisture transfer between food and surrounding atmosphere;

the WVP should be as low as possible, because the level of moisture content inside of the food

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packaging could affect the quality of the products[56]. The WVP of bio-nanocomposite films are

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shown in Table 3. The WVP of the SKZ film was found to be 2.47×10−10 g m−1 s−1 Pa−1and it

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was influenced by exposure to UV-C radiation. It is well known that the polysaccharides are

highly sensitive to moisture and exhibit poor water vapor barrier properties due to their

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hydrophilic character[57]. Importantly, when SKZ solution exposured to UV up to 12 h, the
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WVP decreased significantly to 2.08×10−10 g m−1 s−1 Pa−1, because of the better distribution of

the ZNs in polymer matrix (Figure 2). Furthermore, this decrease in WVP is might be explained
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by the interaction between both biopolymers and non-aggregated ZNs, which lead to decrease
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the hydrophilic character of the biopolymers, and decrease WVP of the UV treated film

specimens. As well as, it seems the prevention of ZN aggregation was effective in decreasing
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WVP by increasing UV exposure time (Figure 4-b). This result is in agreement with previously
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report about the blend of biopolymers [58] and nano reinforcement agent [10, 24] which are

studied for packaging applications.

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Table 3.

3.8 Color

The color of the packaging is an important factor in terms of general appearance and consumer

acceptance [50]. The parameters of L (lightness), a (green–red), b (blue–yellow), ΔE (difference

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between color of white plate and samples), YI (yellowness index) and WI (whiteness index)

values are shown in Table 4. As shown, ΔE and a were decreased by increasing the exposure

time. On the other hand L, b, and YI were decreased, simultaneously. Potentiality of ZNs for

removing dye from textile under UVC light has been proved by Behnajady et al [59]. It seems,

produced H2O2 (Eq. (4)) and induced free radicals production under UV radiation (Eq. (21)) [60]

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were the effective factors to this color changes.

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H2O2 + hν → 2•OH (21)

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Furthermore, there are several factors behind of the color changes under photo-modification such

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as the UV dose and the chemical characteristics (i.e. dye structure, radical scavengers, and pH)

[60]. As well as there are several unknown factors.

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Table 4.
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3.9 UV- Vis spectroscopy

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It is important to protect some light sensitive materials against the effects of light, especially UV
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radiation. Free radicals formation under the UV light can be reduced the quality of food by

destruction of antioxidants, oxidation of lipids, destruction of nutrients, changes in color, and

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formation of off-flavors[61].

The absorption spectra of the SKZU and non-irradiated SKZ nanocomposites in the range of

200–800 nm are shown in Figure 4. A strong absorption peak at 364 nm is clearly observable in

all the specimens which induced by increasing the UV exposure times. These results indicate that

the specimens exhibit an obvious blue-shift phenomenon. The UV-shielding at different UV

regions (i.e. UV-A, UV-B, and UV-C) of the film specimens were induced with increasing

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irradiation time. As shown in Figure 2, after increasing the exposure time from 1 h to 12 h the

homogeneity of ZN in biopolymer matrix was improved. On the other hand, UV absorption of

ZN increase with decreasing the size of the nanoparticle [62]. Hence, it seems UV spectrum of

the ZN based films can be used to compare the distribution and homogeneity of ZN in the matrix

of the polymers.

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Figure 4.

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4. Conclusion

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In this study, bio-nanocomposite films were developed based on SKZ with different UV
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exposure time using the solution casting method. The results showed that the properties of these

films were changed by increasing the UV irradiation time. An incremental change in water
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resistance properties, physical properties, and WVP of SKZ films was observed by increasing
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UV exposure time. Furthermore, UV-Vis spectrum of the film specimens confirmed UV-

protective properties of all specimens against different UV region and induce this properties by
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photo-modification. The homogenous distribution of ZN in the SK matrix contributed to the

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improvement of the bio-nanocomposite properties. Altogether, it also suggests that UV exposure

of the film forming solution was effective on ZN distribution in the matrix of biopolymer. It is
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due to the change of surface properties of ZNs under UV-C light. On the other hand, the

chemical changes in the matrix of the biopolymer by photo-modification are effective on the

packaging properties of the films, too. Therefore, it seem UV can be used as a compatibilizing

method to use ZN in reinforcing biopolymers. Moreover, it seems UV spectrum of the ZN based

biopolymers can be used to compare the homogeneity and the distribution of ZN in the matrix of

biopolymers. But it needs to a more accurate investigation.

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Acknowledgment

This research was supported by University of Zanjan under grant number 9711. The authors are

grateful to the Iran Nanotechnology Initiative Council for providing financial aid for this

research.

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References

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1. Piyada, K., S. Waranyou, and W. Thawien, Mechanical, thermal and structural

US
properties of rice starch films reinforced with rice starch nanocrystals. International

Food Research Journal, 2013. 20(1): p. 439-449.

AN
2. El Miri, N., et al., Bio-nanocomposite films reinforced with cellulose nanocrystals:

Rheology of film-forming solutions, transparency, water vapor barrier and tensile

M

properties of films. Carbohydrate polymers, 2015. 129: p. 156-167.

ED

3. Palmer, R., A. Robinson, and Q. Guo, How nanoscience translates into technology: The

case of self-assembled monolayers, electron-beam writing, and carbon nanomembranes.

PT

ACS nano, 2013. 7(8): p. 6416-6421.

CE

4. Shankar, S., et al., Preparation, characterization, and antimicrobial activity of

gelatin/ZnO nanocomposite films. Food Hydrocolloids, 2015. 45: p. 264-271.

AC

5. Rhim, J.-W., H.-M. Park, and C.-S. Ha, Bio-nanocomposites for food packaging

applications. Progress in Polymer Science, 2013. 38(10): p. 1629-1652.

6. Goudarzi, V., I. Shahabi-Ghahfarrokhi, and A. Babaei-Ghazvini, Preparation of

ecofriendly UV-protective food packaging material by starch/TiO 2 bio-nanocomposite:

23
 ACCEPTED MANUSCRIPT

Characterization. International Journal of Biological Macromolecules, 2017. 95: p. 306-

313.

7. Hassannia-Kolaee, M., et al., Development of ecofriendly bionanocomposite: Whey

protein isolate/pullulan films with nano-SiO 2. International journal of biological

macromolecules, 2016. 86: p. 139-144.

T
8. Hassannia-Kolaee, M., F. Khodaiyan, and I. Shahabi-Ghahfarrokhi, Modification of

IP
functional properties of pullulan–whey protein bionanocomposite films with nanoclay.

CR
Journal of food science and technology, 2016. 53(2): p. 1294-1302.

9. Shahabi-Ghahfarrokhi, I., et al., Green bionanocomposite based on kefiran and cellulose

US
nanocrystals produced from beer industrial residues. International journal of biological
AN
macromolecules, 2015. 77: p. 85-91.

10. Shahabi-Ghahfarrokhi, I., et al., Preparation of UV-protective kefiran/nano-ZnO

M

nanocomposites: Physical and mechanical properties. International journal of biological

ED

macromolecules, 2015. 72: p. 41-46.

11. Rad, F.H., et al., Preparation and Characterization of Pullulan-Soy Protein Concentrate
PT

Blended Film Incorporated With Zataria multiflora and Artemisia biennis Essential Oils.
CE

2016.

12. Shahabi-Ghahfarrokhi, I., et al., Effect of γ-irradiation on the physical and mechanical
AC

properties of kefiran biopolymer film. International journal of biological macromolecules,

2015. 74: p. 343-350.

13. Piermaria, J.A., et al., Films based on kefiran, an exopolysaccharide obtained from kefir

grain: Development and characterization. Food Hydrocolloids, 2009. 23(3): p. 684-690.

24
 ACCEPTED MANUSCRIPT

14. Motedayen, A.A., F. Khodaiyan, and E.A. Salehi, Development and characterisation of

composite films made of kefiran and starch. Food chemistry, 2013. 136(3): p. 1231-1238.

15. Medrano, M., P.F. Pérez, and A.G. Abraham, Kefiran antagonizes cytopathic effects of

Bacillus cereus extracellular factors. International journal of food microbiology, 2008.

122(1): p. 1-7.

T
16. Zubillaga, M., et al., Effect of probiotics and functional foods and their use in different

IP
diseases. Nutrition Research, 2001. 21(3): p. 569-579.

CR
17. Guzel-Seydim, Z.B., et al., Review: functional properties of kefir. Critical reviews in food

science and nutrition, 2011. 51(3): p. 261-268.

18.
US
Vinderola, G., et al., Effects of the oral administration of the exopolysaccharide produced
AN
by Lactobacillus kefiranofaciens on the gut mucosal immunity. Cytokine, 2006. 36(5): p.

254-260.
M

19. Dimitreli, G., et al., Effect of Kefiran and Milk Proteins Addition on the Rheological
ED

Behavior of Glucono-delta-Lactone Induced Milk Gels. Journal of Food Research, 2016.

5(1): p. 121.
PT

20. Kazazi, H., et al., Rheology and microstructure of kefiran and whey protein mixed gels.
CE

Journal of Food Science and Technology: p. 1-7.

21. Angellier, H., et al., Thermoplastic starch− waxy maize starch nanocrystals
AC

nanocomposites. Biomacromolecules, 2006. 7(2): p. 531-539.

22. Goudarzi, V. and I. Shahabi-Ghahfarrokhi, Photo-producible and photo-degradable

starch/TiO 2 bionanocomposite as a food packaging material: Development and

characterization. International Journal of Biological Macromolecules, 2017.

25
 ACCEPTED MANUSCRIPT

23. Reddy, N. and Y. Yang, Citric acid cross-linking of starch films. Food Chemistry, 2010.

118(3): p. 702-711.

24. Babaei-Ghazvini, A., I. Shahabi-Ghahfarrokhi, and V. Goudarzi, Preparation of UV-

protective starch/kefiran/ZnO nanocomposite as a packaging film: Characterization.

Food Packaging and Shelf Life, 2018. 16: p. 103-111.

T
25. Díaz, O., D. Candia, and Á. Cobos, Effects of ultraviolet radiation on properties of films

IP
from whey protein concentrate treated before or after film formation. Food

CR
Hydrocolloids, 2016. 55: p. 189-199.

26. Campos, A., et al., The influence of UV-C irradiation on the properties of thermoplastic

US
starch and polycaprolactone biocomposite with sisal bleached fibers. Polymer
AN
Degradation and Stability, 2012. 97(10): p. 1948-1955.

27. Lee, Y.G., J.-H. Koo, and J. Kim, Influence of cloud fraction and snow cover to the
M

variation of surface UV radiation at King Sejong station, Antarctica. Atmospheric

ED

Research, 2015. 164: p. 99-109.

28. Rhim, J.W., et al., Properties of Ultraviolet Irradiated Protein Films. LWT - Food
PT

Science and Technology, 1999. 32(3): p. 129-133.

CE

29. Sirelkhatim, A., et al., Review on zinc oxide nanoparticles: antibacterial activity and

toxicity mechanism. Nano-Micro Letters, 2015. 7(3): p. 219-242.

AC

30. Ameen, S., et al., An effective nanocomposite of polyaniline and ZnO: preparation,

characterizations, and its photocatalytic activity. Colloid and Polymer Science, 2011.

289(4): p. 415-421.

31. Fan, Z. and J.G. Lu, Zinc oxide nanostructures: synthesis and properties. Journal of

nanoscience and nanotechnology, 2005. 5(10): p. 1561-1573.

26
 ACCEPTED MANUSCRIPT

32. Rodríguez, J., et al., Synthesis and characterization of ZnO nanorod films for

photocatalytic disinfection of contaminated water. Thin Solid Films, 2010. 519(2): p.

729-735.

33. Özgür, Ü., et al., A comprehensive review of ZnO materials and devices. Journal of

applied physics, 2005. 98(4): p. 041301.

T
34. Ludi, B. and M. Niederberger, Zinc oxide nanoparticles: chemical mechanisms and

IP
classical and non-classical crystallization. Dalton Transactions, 2013. 42(35): p. 12554-

CR
12568.

35. Salarbashi, D., et al., Development of new active packaging film made from a soluble

US
soybean polysaccharide incorporating ZnO nanoparticles. Carbohydrate polymers, 2016.
AN
140: p. 220-227.

36. Pera-Titus, M., et al., Degradation of chlorophenols by means of advanced oxidation

M

processes: a general review. Applied Catalysis B: Environmental, 2004. 47(4): p. 219-

ED

256.

37. Sun, R.-D., et al., Photoinduced surface wettability conversion of ZnO and TiO2 thin
PT

films. The Journal of Physical Chemistry B, 2001. 105(10): p. 1984-1990.

CE

38. Bhogale, A., et al., Systematic investigation on the interaction of bovine serum albumin

with ZnO nanoparticles using fluorescence spectroscopy. Colloids and Surfaces B:

AC

Biointerfaces, 2013. 102: p. 257-264.

39. Piermaria, J.A., L. Mariano, and A.G. Abraham, Gelling properties of kefiran, a food-

grade polysaccharide obtained from kefir grain. Food Hydrocolloids, 2008. 22(8): p.

1520-1527.

40. Nielsen, S.S., Food analysis laboratory manual. 2003: Springer.

27
 ACCEPTED MANUSCRIPT

41. Rao, M.A., Rheology of fluid and semisolid foods: principles and applications: principles

and applications. 2010: Springer Science & Business Media.

42. Ojagh, S.M., et al., Development and evaluation of a novel biodegradable film made from

chitosan and cinnamon essential oil with low affinity toward water. Food Chemistry,

2010. 122(1): p. 161-166.

T
43. ASTM, Standard test methods for water vapor transmission of material. 1995, In

IP
Annual book of ASTM: Philadelphia American Society for Testing and Materials.

CR
44. Gennadios, A., C.L. Weller, and C.H. Gooding, Measurement errors in water vapor

permeability of highly permeable, hydrophilic edible films. Journal of food engineering,

1994. 21(4): p. 395-409.

US
AN
45. ASTM. 2002, Annual Book of ASTM: Philadelphia American Society for Testing

and Materials.
M

46. Stuart, B., Infrared Spectroscopy: Fundamentals and Applications. John Wiley &
ED

Sons, 2004: p. 45–93.

PT

47. Kołodziejczak-Radzimska, A. and T. Jesionowski, Zinc oxide—from synthesis to

application: a review. Materials, 2014. 7(4): p. 2833-2881.

CE

48. Bhat, R. and A. Karim, Ultraviolet irradiation improves gel strength of fish gelatin. Food
AC

Chemistry, 2009. 113(4): p. 1160-1164.

49. Delville, J., et al., Solid state photocrosslinked starch based films: a new family of

homogeneous modified starches. Carbohydrate polymers, 2002. 49(1): p. 71-81.

50. Nawapat, D. and W. Thawien, Effect of UV-treatment on the properties of biodegradable

rice starch films. Int Food Res J, 2013. 20: p. 1313-1322.

28
 ACCEPTED MANUSCRIPT

51. Kanmani, P. and J.-W. Rhim, Properties and characterization of bionanocomposite films

prepared with various biopolymers and ZnO nanoparticles. Carbohydrate polymers,

2014. 106: p. 190-199.

52. Ghanbarzadeh, B., et al., Effect of plasticizing sugars on water vapor permeability,

surface energy and microstructure properties of zein films. LWT-Food Science and

T
Technology, 2007. 40(7): p. 1191-1197.

IP
53. Bertolini, A.C., et al., Photodegradation of cassava and corn starches. Journal of

CR
agricultural and food chemistry, 2001. 49(2): p. 675-682.

54. Shahabi-Ghahfarrokhi, I., V. Goudarzi, and A. Babaei-Ghazvini, Production of starch

US
based biopolymer by green photochemical reaction at different UV region as a food
AN
packaging material: Physicochemical characterization. International journal of

biological macromolecules, 2018.

M

55. Bertolini, A., Starches: characterization, properties, and applications. 2009: CRC Press.
ED

56. Phan, T.D., et al., Functional properties of edible agar-based and starch-based films for

food quality preservation. Journal of Agricultural and Food Chemistry, 2005. 53(4): p.
PT

973-981.
CE

57. BenBettaïeb, N., et al., Spectroscopic analyses of the influence of electron beam

irradiation doses on mechanical, transport properties and microstructure of chitosan-fish

AC

gelatin blend films. Food Hydrocolloids, 2015. 46: p. 37-51.

58. Ghanbarzadeh, B., H. Almasi, and A.A. Entezami, Physical properties of edible modified

starch/carboxymethyl cellulose films. Innovative food science & emerging technologies,

2010. 11(4): p. 697-702.

29
 ACCEPTED MANUSCRIPT

59. Behnajady, M., N. Modirshahla, and R. Hamzavi, Kinetic study on photocatalytic

degradation of CI Acid Yellow 23 by ZnO photocatalyst. Journal of hazardous materials,

2006. 133(1): p. 226-232.

60. Ince, N.H., “Critical” effect of hydrogen peroxide in photochemical dye degradation.

Water Research, 1999. 33(4): p. 1080-1084.

T
61. Koutchma, T., Advances in ultraviolet light technology for non-thermal processing of

IP
liquid foods. Food and Bioprocess Technology, 2009. 2(2): p. 138-155.

CR
62. Goh, E., X. Xu, and P. McCormick, Effect of particle size on the UV absorbance of zinc

oxide nanoparticles. Scripta Materialia, 2014. 78: p. 49-52.

US
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Figure captions

Figure 1. a) Scheme of UV irradiation method and activities of nano-ZnO (ZN) under UV ray,

b) Distribution and positioning of ZN between the starch and kefiran chains

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Figure 2. Contact angle of water (A) SEM microstructure of surface (S) and cross section (C) of

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1) starch-kefiran-ZnO (SKZ), 2) UV irradiated SKZ for 1 h, 3) UV irradiated SKZ for 6h,

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and 4) UV irradiated SKZ for 12h by UV-C
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Figure 3. FTIR transmittance spectra of starch-kefiran-ZnO (SKZ) film and UV irradiated films
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with various UV (U) exposure times (1, 6, and 12 h)

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Figure 4. a) UV- visible absorption spectra of starch-kefiran-ZnO (SKZ) film and irradiated

starch-kefiran-ZnO (SKZU) at different UV exposure times (1, 6, and 12 h) films, b)

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water vapor permeability scheme of the non-aggregated and aggregated ZN inside of the
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films

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Table 1. Physical properties of starch-kefiran-ZnO (SKZ) film and UV irradiated films with

various UV (U) exposure times (1, 6, and 12 h) ‡,*

Moisture Moisture Solubility in

Thickness Contact angle
Specimens content absorption water
(mm) (degree)
(%) (%) (%)

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SKZ 0.11±0.00a 27.44±1.87a 11.71±0.10a 29.30±0.75a 95.63±1.57d

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SKZU1 0.12±0.00a 27.63±1.57a 10.95±0.44b 29.05±0.30a 99.55±2.46c

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SKZU6 0.11±0.01a 25.21±1.45b 10.05±0.36c 27.91±0.33b 103.21±1.96b

25.92±1.67b

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SKZU12 0.12±0.01a 9.11±0.74d 26.66±0.17b 107.37±2.43a

‡
Means within each column with the same letters are not significantly different (P<0.05)
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* Data are means ± SD
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Table 2. Physical properties of starch-kefiran-ZnO (SKZ) film and UV irradiated films with

various UV (U) exposure times (1, 6, and 12 h) ‡,*

Tensile energy to
Tensile strength Elongation at break Young’s modulus
Specimens break
(MPa) (%) (MPa)
(MJ)

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SKZ 6.71±0.47b 155.44±3.48a 31.30±1.19b 5.25±0.63a

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SKZU1 7.25±0.33b 145.15±4.03b 32.77±1.63b 5.17±0.44a

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SKZU6 7.44±0.81a 143.12±4.55b 33.55±1.96ab 5.33±0.45a

SKZU12 8.21±0.69a 131.44±3.96c 35.17±2.82a 5.54±0.79a

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‡
Means within each column with the same letters are not significantly different
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(P<0.05)

* Data are means ± SD

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Table 3. WVP and intrinsic viscosity of starch-kefiran-ZnO (SKZ) film and UV irradiated films

with various UV (U) exposure times (1, 6, 12 h) ‡,*

WVP Intrinsic viscosity

Specimens
(×10−10 g m−1 s−1 Pa−1) (cm3/g)

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SKZ 2.47±0.07a 70.66±0.87a

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SKZU1 2.38±0.03a 65.39±0.95b

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SKZU6 2.20±0.06b 60.67±0.66c

SKZU12 2.08±0.09b 56.91±0.78d

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‡
Means within each column with the same letters are not significantly different (P<0.05)

* Data are means ± SD

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Table 4. Hunter color values (L, a, and b), total color difference (∆E), YI (yellowness

index) and whiteness index (WI) of starch-kefiran-ZnO (SKZ) film and UV irradiated films

with various UV (U) exposure times (1, 6, 12 h) ‡,*

Specimens L a b YI ΔE WI

SKZ 95.55±1.17c 1.06±0.50c 1.09±0.45ab 1.62±0.15c 4.54±1.17a 95.29±1.24a

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SKZU1 96.25±1.55b 0.76±0.58b 1.89±0.25ab 2.80±0.21b 3.75±1.03b 95.73±1.11a

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SKZU6 97.18±1.08b 0.77±0.61b 2.31±0.32a 3.39±0.37a 2.84±0.92b 96.27±1.32a

SKZU12 97.97±1.37a 0.45±0.21a 2.73±0.52a 3.98±0.35a 2.59±0.86b 96.56±1.21a

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‡
Means within each column with the same letters are not significantly different (P<0.05)

* Data are means ± SD

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Highlights

 A green method provided to improve the packaging properties of the starch-kefiran film.

 A green method to compatibilizing ZnO as a nano-reinforcement was developed.

 Packaging characterizations of UV irradiated starch-kefiran-ZnO films were investigated.

 UV-protective packaging materials was developed.

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Figure 1
Figure 2
Figure 3
Figure 4