Carbohydrate Polymers 108 (2014) 291–298

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

Green synthesis of colloid silver nanoparticles and resulting

biodegradable starch/silver nanocomposites
Perrine Cheviron a,b,c , Fabrice Gouanvé a,b,c,∗ , Eliane Espuche a,b,c,∗∗
a
Université de Lyon, F-69003 Lyon, France
b
Université Lyon 1, F-69622 Villeurbanne, France
c
CNRS, UMR5223, Ingénierie des Matériaux Polymères, France

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

Article history: Environmentally friendly silver nanocomposite films were prepared by an ex situ method consisting
Received 25 November 2013 firstly in the preparation of colloidal silver dispersions and secondly in the dispersion of the as-prepared
Received in revised form 24 January 2014 nanoparticles in a potato starch/glycerol matrix, keeping a green chemistry process all along the syn-
Accepted 12 February 2014
thesis steps. In the first step concerned with the preparation of the colloidal silver dispersions, water,
Available online 28 February 2014
glucose and soluble starch were used as solvent, reducing agent and stabilizing agent, respectively. The
influences of the glucose amount and reaction time were investigated on the size and size distribution of
Keywords:
the silver nanoparticles. Two distinct silver nanoparticle populations in size (diameter around 5 nm size
Silver
Nanoparticle
for the first one and from 20 to 50 nm for the second one) were distinguished and still highlighted in the
Biopolymer potato starch/glycerol based nanocomposite films. It was remarkable that lower nanoparticle mean sizes
Green chemistry were evidenced by both TEM and UV–vis analyses in the nanocomposites in comparison to the respec-
Starch tive colloidal silver dispersions. A dispersion mechanism based on the potential interactions developed
Morphology between the nanoparticles and the polymer matrix and on the polymer chain lengths was proposed to
explain this morphology. These nanocomposite film series can be viewed as a promising candidate for
many applications in antimicrobial packaging, biomedicines and sensors.
© 2014 Elsevier Ltd. All rights reserved.

1. Introduction inorganic nanofillers to achieve designed functional properties.

Metal nanoparticles incorporated polymers attracted great atten-
Biodegradable polymers from renewable resources have tion because of widened application scope (Clémenson, Léonard,
attracted much attention in recent years (Averous, Faucaonnier, Sage, David, & Espuche, 2008; Nesher, Marom, & Anvir, 2008;
Moro, & Fringant; Peterson, 2000; Hartman, Albertsson, Söderqvist Nimrodh Ananth, Umapathy, Sophia, Mathavan, & Mangalaraj,
Lindblad, & Sjöberg, 2006; Petersson & Oksman, 2006). Renewable 2011; Wang, Wu, & Zhu, 2002; White, Budarin, Moir, & Clark,
sources of polymeric materials offer an alternative for maintain- 2011). Particularly, polymer–silver nanocomposites are promising
ing the sustainable development of economically and ecologically functional materials in fields such as optical, magnetic, electronic
attractive technologies. Indeed, the complete biological degrad- and antimicrobial properties (Duan, Huang, Cui, Wang, & Lieber,
ability of these polymers can contribute to a reduction in the 2001; Huang & Yang, 2004; Huang, Yuan, & Yang, 2004; Jin, Cao,
volume of garbage and the protection of the climate through the Mirkin, Kelley, Schatz, & Zheng, 2001; Lahav, Gabriel, Shipway, &
reduction of the carbon dioxide release. Thus, there is consider- Willner, 1999; Narayanan & El-Sayed, 2005; Sanpui, Murugadoss,
able interest in replacing some or even a large amount of synthetic Prasad, Ghosh, & Chattopadhyay, 2008). Different routes to intro-
polymers by biodegradable materials and in combining them with duce silver nanoparticles into synthetic polymer matrices have
been developed over the past few years (Akamatsu et al., 2000;
Chen, Chen, Serizawa, & Akashi, 1998; Dirix, Bastiaansen, Caseri, &
Smith, 1999; Heilmann & Werner, 1998). In the last decade, many
∗ Corresponding author at: Université de Lyon, F-69003, Lyon, France. efforts have been made in the incorporation of silver nanoparti-
Tel.: +33 04 72 43 12 10. cles into biodegradable polymers for their potential application
∗∗ Corresponding author at: Université de Lyon, F-69003, Lyon, France.
in biotechnology (Murugadoss & Chattopadhyay, 2008). Polysac-
Tel.: +33 04 72 43 27 01.
E-mail addresses: [email protected] (F. Gouanvé), charide polymers, such as chitosan, alginate and starch are the
[email protected] (E. Espuche). most extensively used as host matrices (Bozanic et al., 2011;

http://dx.doi.org/10.1016/j.carbpol.2014.02.059
0144-8617/© 2014 Elsevier Ltd. All rights reserved.
292 P. Cheviron et al. / Carbohydrate Polymers 108 (2014) 291–298

Brayner, Vaulay, Fievet, & Coradin, 2007; Djokovic, Krsmanovic, in this study, it was possible at first to prepare very small silver
Bozanic, McPherson, & Van Tendeloo, 2009; Khan et al., 2013; nanoparticles and then to keep a fine dispersion of these nanopar-
Vigneshwaran, Nachane, Balasubramanya, & Varadarajan, 2006; ticles within the biodegradable polymer matrix. This is a promising
Vimala, Murali Mohan, Samba Sivudu, Varaprasad, Ravindra, 2010; item for further functional fields such as medical applications and
Wei, Sun, Qian, & Ye, 2009). The general method consists in the packaging.
dispersion of already prepared colloidal nanoparticles (AgNPs) in
the polymer matrix solution (Lim & Ast, 2001). This method is 2. Experimental
often referred to as the evaporation method since the polymer
solvent is evaporated from the reaction mixture after AgNPs dis- 2.1. Materials
persion. In all cases, the performance level of the nanocomposites
depends on the controlled distribution of the uniformly shaped and For colloidal silver nanoparticles preparation, AgNO3 (ACS
sized nanoparticles. Improved properties are generally obtained reagent > 99.0%) was purchased from Aldrich and used as silver pre-
when small dispersed nanodomains are obtained (Wiley, Sun, & cursor. d(+)-Glucose from Merck was used as reducing agent and
Xia, 2007). Therefore, the controllable synthesis of AgNPs is the soluble starch, supplied from Aldrich, was introduced as nanopar-
first key challenge to achieve their better applied characteristics. ticles stabilizer. For the nanocomposite films elaboration, native
Colloidal silver nanoparticles can be prepared by physical, bio- potato starch with a weight ratio of amylopectin to amylase equal to
logical and chemical methods. The chemical approach is mostly 77/23 was purchased from Sigma and glycerol (99% purity-supplied
used and consists in the treatment of silver salts with a chemical from Aldrich) was used as plasticizer. In all cases, distilled water
reducing agent, such as hydrazine, ethylenediaminetetraacetic acid was used as solvent.
and above all sodium borohydride (Bright, Musick, & Natan, 1998;
Evanoff & Chumanov, 2004; Merga, Wilson, Lynn, Milosavljevic, & 2.2. Preparation of the colloidal silver nanoparticles
Meisel, 2007; Wang, Efrima, & Regev, 1998). However, most of these
reducing agents are considered as non-environmentally friendly In a typical experiment, 85 mg of soluble starch was dissolved
component. Increasing awareness about the total elimination or at in 25 mL of distilled water at 85 ◦ C for 30 min. 10 mL of a 6.10−2 M
least the minimization of the generated waste has led researchers silver nitrate was added into 25 mL of hot aqueous solution of sol-
to focus on alternative synthesis routes. Recently, efforts have been uble starch under vigorous stirring away from light. Then 15 mL of
made for developing green methods to prepare silver nanoparti- a glucose solution (0.06 M or 0.12 M) was added into the reactive
cles (Anastas & Warner, 1998; Huang & Yang, 2004; Raveendran, system, which was held at 85 ± 0.5 ◦ C with 700 rpm stirring. A ther-
Fu, & Wallen, 2003). The preparation of AgNPs via a green chem- mostated oil bath and a magnetic stirrer were used to maintain a
istry process should be evaluated from three specific aspects: the constant temperature and constant stirring throughout the reac-
choice of the solvent medium, the choice of a non-toxic reducing tion process. The molar fractions of AgNO3 /glucose used were 1:0,
agent and finally the choice of environmental friendly materials 1:1.5 and 1:3. The obtained colloidal suspension was cooled down
for the stabilization of the silver nanoparticles. Water is generally at room temperature after different reaction times (between 1 and
used as the environmental friendly solvent. Saccharide molecules 72 h) and characterized. To avoid any photochemical reactions, all
such as aldehyde are generally used for the reduction of silver ions. colloidal dispersions were kept in a dark place.
Monosaccharide like glucose, galactose and fructose and disaccha-
ride like maltose, lactose have been reported to lead to AgNPs of 2.3. Preparation of the nanocomposite films
controllable sizes (Panacek, Kvıtek, Prucek, Kolar, & Vecerova, 2006;
Mehta, Chaudhary, & Gradzielski, 2010). Glucose is one of the most The film preparation consisted of the dissolution of potato starch
widely-used green reducing agent due to its chemical reaction rate in the presence of glycerol (weight ratio 85:15) in distilled water
which allows a compromise between the number of nuclei created at a concentration of 3 wt%. The solutions were heated to the
and the rate of growth of the silver nanoparticles (Mehta et al., gelatinization temperature (85 ◦ C) and continuously stirred at this
2010; Panigrahi, Kundu, Ghosh, Nath, & Pal, 2004). Polyethylene temperature for 3 h. Then, an appropriate amount of colloidal sil-
glycol and polysaccharides such as starch, chitosan and heparin ver nanoparticles was added and further stirred for 2 min at room
are reported to stabilize silver nanoparticles (Fanta, Kenar, Felker, temperature. The obtained mixture was poured into polystyrene
& Byars, 2013; Shameli et al., 2012; Sharma, Yngard, & Lin, 2009). Petri dishes and water evaporation was carried out at ambient
In some cases, polysaccharides serve as both reducing and capping temperature away from light during at least four days. The theo-
agent (Huang & Yang, 2004). In a nanocomposite approach, the retical silver nanoparticles content in the films was 2 wt% and was
choice of the capping agent is crucial to allow a good incorporation expressed with respect to the total matter content including glyc-
and final dispersion of AgNPs in the polymer matrix. erol. Neat matrices were prepared by using the same experimental
Among all the studies related with silver nanoparticles, very few conditions. In all cases, the films were conditioned in dark place at
works have been concerned with the whole steps of the preparation 25 ◦ C between two aluminum sheets before characterization. The
of biodegradable silver/polymer nanocomposites from green pro- thickness of the films was approximately 60 ␮m ± 5 ␮m.
cess including the synthesis of colloidal silver nanoparticles, their
incorporation in a biodegradable polymer matrix and finally the 2.4. Characterization methods
characterization of the obtained nanocomposites. In the present
paper, the synthesis of colloidal silver nanoparticles via a green Different techniques were used to obtain complementary
chemistry process is reported. Water, glucose and soluble starch information about the size and the morphology of the silver
have been used as solvent, reducing agent and stabilizing agent, nanoparticles in the colloidal suspensions and in the nanocompos-
respectively. A kinetic study of the AgNPs formation was performed ite films.
as a function of the concentration of reducing agent. The synthe-
sis process was studied by using complementary techniques and 2.4.1. Ultraviolet–visible absorption spectroscopy (UV–vis)
the size and size distribution of the AgNPs were characterized. To follow the formation of silver nanoparticles and to inves-
Starch based nanocomposite films were then prepared from the tigate the influence of the embedded silver nanoparticles in the
synthesized silver nanoparticles and their morphology was char- matrix, Ultraviolet–visible (UV–vis) absorption studies were per-
acterized. It was shown that by the green chemistry approach used formed on the colloidal silver nanoparticle dispersions and on the
 P. Cheviron et al. / Carbohydrate Polymers 108 (2014) 291–298 293

1.5 t=72 h 1.5 t=72 h 1.5 t=72 h

t=48 h t=48 h t=48 h

t=24 h t=24 h t=24 h

t=15 h t=15 h t=15 h

Absorbance

Absorbance

Absorbance
1 t=6 h 1 t=6 h 1 t=6 h

t=1 h t=1 h t=1 h

t=0 h t=0 h
t=0 h

0.5 0.5 0.5

0 0 0
300 350 400 450 500 550 600 650 700 300 350 400 450 500 550 600 650 700 300 350 400 450 500 550 600 650 700

Wavelength (nm) Wavelength (nm) Wavelength (nm)

(a) (b) (c)

Fig. 1. UV–vis spectral evolution of silver nanoparticles using different AgNO3 /glucose molar ratio: (a) 1/3, (b) 1/1.5 and (c) 1/0.

different nanocomposite films with a Perkin Elmer Lambda 750 solution. The most characteristic evidence of silver nanoparticles
spectrophotometer in the wavelength range of 200–700 nm. formation is the surface plasmon resonance (SPR) bands observable
in the 350–600 nm region (Burda, Chen, Narayanan, El-Sayed, 2005;
2.4.2. Transmission electron microscopy (TEM) Daniel & Astruc, 2004; Henglein, 1989). The shape of the spectra
Size analysis of silver nanoparticles was carried out with a gives preliminary information about the size and the size distri-
Philips CM120 electron microscope with an accelerating voltage of bution of the silver nanoparticles (Harada, Inada, & Nomura, 2009;
120 kV. Two different methods were used to perform the colloidal Rafey, Shrivastavaa, Iqbal, & Khan, 2011). The UV–visible absorption
silver nanoparticle dispersion analysis. In a first step a droplet of spectra of silver nanoparticle dispersion at different reaction times
the aqueous dispersion was placed on Formwar coated grids. Then, for AgNO3 /glucose molar ratio of 1:3 and 1:1.5 are represented in
in a second step, the colloidal droplets were dried either directly at Fig. 1a and b, respectively. In presence of the reducing agent, what-
room temperature for 10 min or after using a paper towel to obtain ever the amount of glucose in the mixture, the intensity of the SPR
a thinner deposit. For the nanocomposite films analysis, samples peak increases with the increasing reaction time indicating a con-
were cut with a cryo-ultramicrotome at −90 ◦ C with a Reicher tinued reduction of the Ag+ ions. It can be also noticed that the
Ultracut S instrument equipped with a diamond knife to obtain absorption peak obtained for the reaction mixture prepared from
ultrathin sections of about 60 nm thicknesses. The samples were AgNO3 /glucose molar ratio equal to 1:3 is characterized by a sym-
also placed on Formwar coated grids. For each analysis, low electron metrical shape whatever the reaction time. For the AgNO3 /glucose
beam intensity was used and short time of exposure was performed molar ratio of 1:1.5, the absorption peak has an asymmetric shape
to avoid any evolution of the samples during their exposure to the manifested by a widening toward long wavelengths. This widen-
electron beam. The average diameter and size distribution of the ing at longer wavelengths indicates the presence of nanoparticle
nanoparticles were determined by the ImageJ Software based on clusters (Huang et al., 1996; Patakfalvi, Viranyi, & Dekany, 2004).
the data of an average of 50–500 nanoparticles. In order to evidence the role of glucose in the silver reduction
process, the same experiments were also carried out without any
3. Results and discussion glucose. The corresponding UV–vis absorption spectra at different
reaction times are presented in Fig. 1c. The appearance of the SPR
3.1. Silver nanoparticles synthesis peak shows that the reduction of silver ions also takes place even
if no glucose was added to the mixture. Thus, soluble starch has
Most of the studies based on green synthesis of silver nanopar- a double role: the one of a stabilizer and the one of a reducing
ticles using glucose and soluble starch as reducing agent and agent. However, although the intensity of the SPR peak increases
stabilizer, respectively considered a unique reaction time to pre- with the increase of the reaction time, the peak intensity remains
pare the nanoparticles. However, the chosen reaction time varied lower compared to that one obtained for the mixture containing
arbitrarily from 1 to 20 h depending on the authors (Aguilar- glucose, at the same reaction time. Moreover, the width of the
Mendez, San Martín-Martínez, Ortega-Arroyo, Cobián-Portillo, & peak also increases toward higher wavelengths as a function of the
Sánchez-Espíndola, 2011; Gao, Wei, Yan, & Xu, 2011; Raveendran reaction time. The wavelength at the maximum of the absorbance
et al., 2003; Shervani & Yamamoto, 2011; Singh, Sinha, & Mandal, (max ) as a function of the reaction time is represented in Fig. 2 for
2009). In the present work, the reaction processes were studied the different AgNO3 /glucose molar ratios. When glucose is added
for increasing reaction times in the range from 1 to 72 h at 85 ◦ C. in the reaction mixture, max is constant (422 ± 3 nm) and does
A series of reaction mixtures were prepared by keeping constant not change as a function of the reaction time, independently of
in each system the concentration of silver precursor (6.10−4 M) the glucose amount. In absence of glucose, the values of max are
as well as the concentration of soluble starch used as stabilizer also constant but slightly higher (427 ± 2 nm) than those obtained
(0.17 wt%) and by varying the amount of reducing agent. The molar for the mixtures containing glucose for reaction times lower than
ratios of AgNO3 /glucose were 1:3 and 1:1.5 respectively. To investi- 24 h. Up to this time, max gradually increases as the reaction time
gate the role of the reducing agent in the reaction process, reaction increases. After 72 h of reaction, max is around 450 nm. Fornasiero
mixtures were also prepared in the same conditions without glu- et al. have demonstrated that an increase in average size of the sil-
cose (AgNO3 /glucose molar ratio: 1:0). Different spectroscopic and ver nanoparticles results in a shift of the absorption peak toward
microscopic techniques are routinely used to investigate the forma- higher wavelengths (Fornasiero & Grieser, 1991). According to this
tion of silver nanoparticles (Anastas & Warner, 1998; Mehta et al., work and to our results, an increase of the nanoparticle size should
2010; Raveendran et al., 2003; Shameli et al., 2012). UV–visible be expected. The rate of Ag+ ion reduction was also determined
spectroscopy is one of the most important techniques to ascer- for each AgNO3 /glucose ratio by examining the time dependence
tain the formation and stability of silver nanoparticles in aqueous of maximal absorbance (Amax ). The corresponding curves are
294 P. Cheviron et al. / Carbohydrate Polymers 108 (2014) 291–298

460 According to the classical crystallization theory, in presence of glu-

1/0 cose, a large number of nuclei are formed and the obtained particle
1/1.5 sizes are smaller and monodisperse. On the contrary, in the absence
1/3 of glucose, a slow nucleation process is obtained leading to a small
440 number of nuclei which grow and reach large polydisperse sizes
λmax (nm)

(Khan, Al-Thabaiti, & Al-Nowaiser, 2012; Patakfalvi et al., 2004).

So, for the molar ratios of AgNO3 /glucose under consideration in
this study, we can conclude that glucose molecules tend to speed
420 up the silver ions reduction and allow to limit the presence of silver
nanoparticle clusters.
The sizes and morphology of synthesized silver nanoparticles
were also imaged using Transmission Electron Microscope (TEM).
400 Analyses were performed first by placing droplets of the undi-
0 10 20 30 40 50 60 70 80 90 luted dispersion on Formwar coated grids. Then, the grids were
Time (hour)
dried at room temperature for 10 min. Colloidal nanoparticle dis-
Fig. 2. Evolution of the wavelength at the maximal absorbance as a function of
persion analyses were done for different reaction times (1, 6,
reaction time. 24, and 48 h) and for the three AgNO3 /glucose molar ratios (1:3,
1:1.5 and 1:0). Characteristic examples of the obtained images are
represented in Fig. 3. In presence of glucose, the curves clearly shown in Fig. 4 for 1, 24 and 48 h of reaction. In a general point
reveal that the Amax value rapidly increases during the initial period of view, the nanoparticles are isotropic and appear to be spher-
of the reaction and then reaches a constant value. The time to reach ical. Two populations of nanoparticles can be distinguished. The
the plateau does not depend on the amount of reducing agent in first one is composed of larger nanoparticles that can be clearly
the mixture. However, without glucose, the absorbance continu- distinguished due to their high enough phase contrast and the
ously increases as a function of the reaction time. The curves of second one of very small nanoparticles (diameter less than 5 nm)
absorbance time dependence represented in Fig. 3 were fitted by a which are characterized by a very low phase-contrast. In a first
first order rate equation according the following relationship: step, a quantitative analysis of the nanoparticles sizes was per-
formed on the first highest size nanoparticles population. For all
At = A∞ (1 − exp(−k.t)) images, nanoparticle sizes were determined using the ImageJ Soft-
ware based on the data of at least 50 measured nanoparticles for
where At is the absorbance at time t, A∞ the absorbance at very long
each picture. The histograms describing the dispersity in size dis-
time and k the first order-rate constant. For each system, the values
tribution are also shown in Fig. 4. The obtained mean average
of A∞ and k are listed in Table 1 and the obtained theoretical curve
nanoparticle size (d) and associated standard deviation () val-
is plotted in dotted line in Fig. 3. We can observe that the first order
ues are given in Table 1. The nanoparticle size d as a function of
rate equation is convenient and well describes the experimental
the reaction time for the different mixtures is also represented in
kinetics. For the different reaction media containing glucose, what-
Fig. 5. For the AgNO3 /glucose molar ratio equal to 1:3, the average
ever the amount of glucose, the same value of k (k = 1.5 × 10−3 s−1 )
particle size is around 20 nm and does not change as a function
is obtained indicating a similar kinetic behavior of the reactions.
of time. More than 80% of the nanoparticles are in the size range
The overall reaction mechanism is shown below:
from 10 to 30 nm. For long reaction time (48 h), only few parti-
2Ag+ + H2 O + CH2 OH − (CHOH)4 cles over 30 nm are observed. Concerning AgNO3 /glucose molar
ratio of 1:1.5, for reaction times lower than 6 h, the average size
− (CHO) → 2Ag + 2H+ + CH2 OH − (CHOH)4 − (COOH)
and the size distribution are equivalent to those obtained with
the higher AgNO3 /glucose ratio (1:3). For time up to 6 h, the aver-
However, in the absence of glucose in the mixture, the kinetic age size increases and is around 30 nm. More than 40% of the
rate constant is 50 times lower. As explained by Gao et al. (2011), particles are bigger than 30 nm. Without glucose, the mean size
the aldehyde terminal of soluble starch can reduce silver nitrate and the size distribution are equivalent to those obtained in the
while the presence of OH and O groups of starch stabilize presence of glucose but only for very short reaction times (less
the silver nanoparticles. Furthermore, the value of A∞ increases than 1 h of reaction). Then, the mean particle size increases as
as the amount of glucose in the reaction mixture increases. the reaction time increases. For 48 h of reaction, the mean parti-
cle size is around 47 nm and more than 60% of the nanoparticles
are bigger than 30 nm. The quantitative data obtained from the
TEM images are in good agreement with the previous qualitative
1.5 conclusions drawn from the analysis of the UV–visible absorp-
tion spectra. However and as already discussed, very small-sized
nanoparticles (less than 5 nm) could also co-exist with high-sized
nanoparticles. It was difficult to perform quantitative analyses of
1
Absorbance

these AgNPs due to their low phase contrast. In order to better

observe this population of very small nanoparticles, the conditions
used to prepare the samples for TEM observation were modified.
1/0
0.5 Instead of drying directly the colloidal droplets at room tempera-
1/1.5
1/3
ture for 10 min, a paper towel was used to obtain thinner deposit
Theoretical curve and allow a better observation of the very small objects under
the electron beam. A characteristic image of the colloidal solu-
0 tion obtained for 24 h of reaction time with a AgNO3 /glucose molar
0 10 20 30 40 50 60 70 80 90
Time (hour)
ratio of 1:3 is shown in Fig. 6. Numerous very small nanoparti-
cles which are in a size range from 2 to 5 nm can be observed in
Fig. 3. Evolution of the maximal absorbance as a function of reaction time. addition to larger nanoparticles. However, the contrast remains
 P. Cheviron et al. / Carbohydrate Polymers 108 (2014) 291–298 295

Table 1
Values of the parameters characterizing the first-order rate kinetic and corresponding mean silver nanoparticles size (d̄) and associated standard deviation () obtained from
the colloidal dispersion.

AgNO3 /Glucose ratio Kinetic Nanoparticles size (nm)

A∞ k × 103 (s−1 ) 1h 6h 24 h 48 h

d̄  d̄  d̄  d̄ 

1/3 1.38 1.51 23 9 21 9 20 12 20 7

1/1.5 1.05 1.52 21 9 21 13 29 19 33 19
1/0 0.97 0.03 21 12 36 19 39 22 47 32

d̄: mean nanoparticles size; :standard deviation.

Fig. 4. Transmission electron micrographs for the colloidal dispersions for different reaction time and different AgNO3 /glucose molar ratio.
296 P. Cheviron et al. / Carbohydrate Polymers 108 (2014) 291–298

60 2
1/0 1/1.5 1/3
1/3
50 1/1.5
1.5 1/0
Neat matrix
40
Average size (nm)

Absorbance
30 1

20
0.5

10

0 0
0 10 20 30 40 50 60 300 350 400 450 500 550 600 650 700

Time(h) Wavelength (nm)

Fig. 5. Evolution of the average size of the silver nanoparticles as a function of Fig. 7. UV–visible absorption spectra of nanocomposite films containing silver
reaction time. nanoparticles synthesized after 48 h of reaction time using different AgNO3 /glucose
molar ratio and reference matrix.

too low to perform a quantitative analysis of this nanoparticle

population. when they are embedded in the polymer matrix. Soluble starch
chains which are used as capping agent can prevent more effi-
3.2. Silver nanocomposite films ciently cluster formation of the silver nanoparticles when they are
dispersed in the potato starch polymer matrix.
Nanocomposite films containing 2 wt% of silver were prepared The dispersion of silver nanoparticles within the plasticized
by mixing an aqueous solution of potato starch and glycerol potato starch matrix was further analyzed using Transmission Elec-
with the colloidal dispersions prepared from the three distinct tron Microscopy. The neat matrix was also imaged to be sure that
AgNO3 /glucose molar ratios for 48 h of reaction respectively. neither the potato starch matrix nor the glycerol plasticizer cre-
According to UV analysis, for this reaction time, the reduction of ates any texturing which could alter the observation of the silver
silver ions is considered to be achieved when glucose is present nanoparticles. Fig. 8a confirms that the plasticized neat matrix is
in the mixture. The aqueous solutions containing potato starch, homogenous. Fig. 8b–d shows TEM images of the nanocompos-
glycerol and AgNPs were cast and the water solvent was evapo- ite films prepared from the silver colloidal dispersion synthesized
rated at room temperature away from light. The obtained films from AgNO3 /glucose molar ratios of 1:3, 1:1.5 and 1:0, respec-
were handleable and not brittle. Furthermore, the colorless neat tively. The nanoparticles are well distributed in the polymer matrix.
matrix film changed to light-orange color when silver nanoparticles Moreover, due to a sufficiently high phase contrast between the
were introduced. The UV–visible absorption spectra of the differ- polymer matrix and the metal nanoparticles, it is possible this time
ent nanocomposites and the neat matrix are presented in Fig. 7. to distinguish two different size populations in all nanocompos-
The absorption values were normalized taking into account the real ites and to perform a quantitative analysis of the nanoparticle sizes
thickness of the films. Whatever the molar ratio of AgNO3 /glucose, by Image J software, considering now all the nanoparticles. The
the absorbance SPR peak confirms the presence of silver nanopar- first identified nanoparticle population corresponds to the silver
ticles. The absorption peaks have a quite symmetrical shape and nanoparticles of bigger size which were already identified in the
the wavelength of maximal absorbance is at 418 nm. It could also colloidal aqueous silver dispersion. The sizes of these nanoparticles
be noted that the widths of the absorption peaks of the different are in the same range order as that obtained for the respective col-
nanocomposites films are lower than those obtained for the asso- loidal dispersions. The second population which corresponds to the
ciated colloidal dispersions. This difference of absorption peaks smallest nanoparticles represents 60% of the numbered nanoparti-
width and the shift of the absorption peak toward lower wave- cles in the nanocomposite films obtained from the silver colloidal
length value can be explained by a change of organization of the dispersion synthesized in absence of glucose. It represents 90%
silver nanoparticles when they are dispersed in water solvent and of the numbered nanoparticles for the nanocomposites prepared

Fig. 6. Transmission electron micrographs for the colloidal dispersion for 24 h of reaction and AgNO3 /glucose molar ratio 1/3 using the towel paper method for the droplet
drying.
 P. Cheviron et al. / Carbohydrate Polymers 108 (2014) 291–298 297

Fig. 8. Transmission electron micrographs for the nanocomposite films containing silver nanoparticles synthesized after 48 h of reaction time using different AgNO3 /glucose
molar ratio: (a) neat matrix, (b) 1/3, (c) 1/1.5 and (d) 1/0.

from the silver colloidal dispersion synthesized in the presence of lower mean size of the nanoparticles evidenced by both TEM and
glucose. The average size of these particles is around 5 nm and it UV–vis analyses, for the nanocomposites in comparison with the
is similar whatever the amount of glucose used during the silver respective colloidal silver dispersions was mainly assigned to the
synthesis step. The observation of an important fraction of very interactions between the soluble starch and the plasticized starch
small nanoparticles dispersed within the plasticized starch poly- matrix which favored the insertion of the starch polymer chain
mer matrix confirms firstly that very small sized nanoparticles between the nanoparticles allowing then these long polymer chains
could be synthesized via the green synthesis process that was pro- to split-off the silver nanoparticles clusters during the film process
posed in this study and secondly that using plasticized potato starch formation.
as nanocomposite polymer matrix should prevent from aggrega- From all our results, it could be concluded that by the green
tion phenomena. It was especially remarkable to evidence by both chemistry ex situ approach used in this study, it was possible at
TEM observation and UV–visible analysis that the nanoparticle dis- first to prepare very small silver nanoparticles and then to obtain
persion was improved in the polymer matrix in comparison to the a fine dispersion of these nanoparticles within the biodegradable
colloidal dispersion. As discussed previously, the presence of solu- starch/glycerol polymer matrix. These nanocomposite films series
ble starch in the colloidal dispersion allows a better dispersion of can be viewed as promising candidates for many applications in
the silver nanoparticles in the plasticized starch matrix due to their antimicrobial, biomedicines and sensors. Further work will be dealt
similar chemical structure and the high molar mass polymer chains with the preparation of biodegradable silver nanocomposite films
of potato starch. directly from an in situ growing process of the silver nanoparticles
via a green chemistry approach.

4. Conclusions
Acknowledgments
Colloidal silver nanoparticles were prepared at 85 ◦ C via a green
The authors are grateful to Ministry of Higher Education and
chemistry process by using water, glucose and soluble starch as sol-
Research for the financial support. They gratefully acknowledge
vent, reducing agent and stabilizing agent respectively. Reactions
Pierre Alcouffe and the Centre Technologique des Microstructures
were carried out for AgNO3 /glucose molar ratios equal to 1:0, 1:1.5
of University of Lyon 1 for TEM photomicrographs.
and 1:3. Kinetics analyses of the silver nanoparticle formation were
performed using ultraviolet-visible absorption spectroscopy and a
first order kinetic was determined. When glucose was added in the Appendix A. Supplementary data
reaction mixture, the reduction of silver ions was considered to be
achieved after 48 h of reaction. At the AgNO3 /glucose levels of 1:3 Supplementary data associated with this article can be found,
and 1:1.5, the kinetic rate constant was similar (k = 1.5 10−3 s−1 ) in the online version, at http://dx.doi.org/10.1016/j.carbpol.2014.
whereas it was 50 times lower in absence of the reducing agent. 02.059.
Spherical silver nanoparticles were evidenced from TEM analy-
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