Progress on Chemistry and Application of Chitin and Its ...

, Volume XV, 2010 63

SYNTHESIS OF SILVER NANOPARTICLES IN A CHITOSAN SOLUTION
Zofa Modrzejewska, Roman Zarzycki, Jan Sielski
Faculty of Process and Environmental Engineering
Technical University of d,
ul. Wlczaska 175, 90-924 d, Poland
e-mail: zmodrzej@wipos.p.lodz.pl
Abstract
In the present study silver nanoparticles were obtained using the ability of chitosan to form
chelate compounds with metal ions and taking advantage of its reducing properties. The reduction
process was conducted at the temperature 60C or applying radiation from 60Co cobalt source.
A preliminary characteristic which allows us to realize the size of formed silver nanoparticles
was made on the basis of the measurements of UV absorption and dynamic light dispersion.
Key words: chitosan salt, nanoparticle, silver.
Progress on Chemistry and Application of Chitin and Its ..., Volume XV, 2010 64
Z. Modrzejewska, R. Zarzycki, J. Sielski
1. Introduction
Currently, nanotechnologies and nanomaterials belong to research priorities. Na-
no-sized substances such as fullerens, nanotubes, nanocolloids, self-organizing particles,
surface structures, nanocrystals, nanosons and nanocapsules are characterized by developed
wall surfaces and higher atom availability. Surface development causes that the contribution
of surface atoms increases, reactivity and properties change. Thanks to nanotechnologies,
materials of very high strength, unusually light, changing color depending on the environ-
ment color and self-purifying can be produced. Owing to their unique properties, nano-
products fnd wide-ranging applications, from medicine and pharmaceutics to electronic
industry, optics and environmental protection.
Nanoparticles are obtained by two methods: top-down, which consists in com-
minution of the material to very fne particles, and bottom-up, which includes chemical
synthesis of nanomaterials (mainly the sol-gel and precipitation methods), forced and free
self-organization (carbon fullerens and nanotubes).
In order to obtain a product in the form of nanopowder, either calcination or drying
of the material to very fne particles, and bottom-up, which includes chemical synthesis of
nanomaterials (mainly the sol-gel and precipitation methods), forced and free self-organiza-
tion (carbon fullerens and nanotubes).
In order to obtain a product in the form of nanopowder, either calcination or drying
of the formed gel is carried out at elevated temperature. Nanoparticles can also be obtained
in a solution. However, they form clusters very quickly and require the use of stabilizers
in the form of surfactants. Nanoparticles are also stabilized by polymers (polyacrylnitrile,
polyvinyl pyrrolidone, polyvinyl alcohol, starch, gelatin, heparin, carboxymethylcellulose
and chitosan), and also by high molecular fatty acids.
A subject of the present study are silver nanoparticles. Researchers are interested
in them mainly because of their unique bactericidal and fungicidal properties. The cidal
activity of silver nanoparticles depends on particle dimensions. The mechanism of silver
nanoparticle activity depends on the type of microorganisms. In fungi it covers disturbance
of the natural water balance by blocking the ability of fungi to bind water molecules. In
the case of viruses this mechanism is reduced to the elimination of their ability of catalytic
decomposition of a lipid-protein substrate.
There are also several ways in which silver nanoparticles can act on a bacterial
cell.
n Silver nanoparticles, which are characterized by high electric conductivity, adhere to a
bacterial cell membrane and disturb the natural gradient of electric potential which deter-
mines the fow of matter and energy indispensable for bacteria life.
n Nanosilver also penetrates protoplasm through capillaries in the cell membrane, causing
disturbances in the activity of mitochondria and cell nucleus.
Progress on Chemistry and Application of Chitin and Its ..., Volume XV, 2010 65
Synthesis of Silver Nanoparticles in a Chitosan Solution
n Silver ions in microbial cells are also bound with thiol groups (-SH) which results in
protein deactivation. Bacterial growth is also inhibited by atomic oxygen adsorbed on
silver surface.
This diversifed action of silver on bacteria causes that so far they have not been
able to form an effcient defense mechanism and practically there are no strains resistant to
silver bactericidal action [1 - 7].
Silver nanoparticles are usually obtained by reducing silver nitrate by sodium
borohydride (NaBH
4
), EDTA or radiation. Photochemical reduction, microwave, electron
beam, micelle, reverse micelle, microemulsion, lamellar liquid crystal, aerosol spraying and
laser radiation are also applied [8 - 20].

In water solutions silver hydrosols agglomerate very quickly and require a stabi-
lizer in the form of a surface-active compound. To stabilize silver nanoparticles also natural
and synthetic polymers are used. Kapoor (1998) stabilized the photochemically reduced sil-
ver particles with polyvinyl pyrrolidone (PVP), gelatin and carboxymethylcellulose (CMC),
while Liu (2001) synthesized polyacrylnitrile-silver nanocomposites using radiation. It is
also known that chitosan can be used for this purpose. Silver nanoparticles were obtained
from silver nitrate introduced to chitosan salt which additionally contained alcohol (isopro-
panol) enabling the growth of free radicals induced by irradiation and reduction of ionic
form of the metal [21 - 27].
2. Experimental results
In the present study silver nanoparticles were obtained using the ability of chitosan
to form chelate compounds with metal ions and taking advantage of its reducing properties.
The reduction process was conducted at the temperature 60C or applying radiation from
60
Cd cobalt source.
Sorption abilities of chitosan and a possibility to form chelate compounds are stud-
ied both in literature and in our own research projects. It follows from them clearly that the
capacity and mechanism of sorption depend on the form of chitosan, which is illustrated in
Figs. 1 and 2. To compare the possibility of silver ion removal by means of chitosan in dif-
ferent forms, studies of Ag
+
adsorption by hydrogel granules and chitosan salt were carried
out simultaneously, as discussed in [28]. The chitosan salt was 4% chitosan acetate, i.e. a so-
lution from which chitosan hydrogel granules were produced. The experiments were carried
out according to the following procedure: to silver solutions with appropriate concentration
of 10 mg Ag/l and 50 mg Ag/l chitosan acetate at pH = 4 was added. The amount of chitosan
in the added salt was equal to the quantity of chitosan present in the hydrogel. Results are
shown in Figures 1 and 2.
It follows from the above data that neutralized chitosan in the form of hydrogel is
characterized by higher sorption capacity. Sorption ability of salt solutions is lower by half.
This is probably a result of the presence of practically all protonated amino groups which
are reactive towards anions, while binding of cations is possible only after deprotonation of
the amino group and it proceeds most probably in two stages:
Progress on Chemistry and Application of Chitin and Its ..., Volume XV, 2010 66
Z. Modrzejewska, R. Zarzycki, J. Sielski
RNH
3
+
RNH
2
+ H
+
+ +
+
n
) RNH ( M RNH n M
2 2
M
+
+ nRNH
2
M(RNH
2
)
n
+
Jangs experiments with copper show that at pH lower than 5.3, binding of cations
with glucosamine molecules proceeds according to the hanging droplet model the metal
ion is bound with one amino group, while at pH above 5.8, like in the bridge model the
metal ions are bound with several amino groups of the same chain or of different chains.
Assuming the same mechanism of reaction for silver, the complexing of chitosan in the form
of hydrogel causes in a certain sense cross-linking of the structure, while salt complexing
offers a possibility of placing silver outside the molecule and enables a reaction with the
products of water decomposition induced by radiation, probably according to the following
reaction patterns:
H
2
O e
aq
-
,

OH, H

, H
3
O
+
, H
2
, H
2
O
2
Ag
+
+ [C
6
H
11
O
4
N]
n
[C
6
H
11
O
4
N]
n
Ag
+
[C
6
H
11
O
4
N]
n
Ag
+
+ e
aq
-
[C
6
H
11
O
4
N]
n
Ag
0
[C
6
H
11
O
4
N]
n
Ag
0
+ Ag
+
[C
6
H
11
O
4
N]
n
Ag
2
+
[C
6
H
11
O
4
N]
n
Ag
2
+
+ Ag
0
[C
6
H
11
O
4
N]
n
Ag
m
+
[C
6
H
11
O
4
N]
n
Ag
m
+
+ e
aq
-
[C
6
H
11
O
4
N]
n
Ag
m
0
[C
6
H
11
O
4
N]
n
+

OH [C
6
H
11
O
4
N]
n-1
[C
6
H
11
O
4
N]

+ H
2
O
[C
6
H
11
O
4
N]
n
Ag
m
+
+ [C
6
H
11
O
4
N]
n-1
[C
6
H
11
O
4
N]

+ H
2
O [C
6
H
11
O
4
N]
n
Ag
m
0
+
[C
6
H
11
O
4
N]
n-1
[C
6
H
11
O
4
N] + H
3
O
+
Silver nanoparticles were obtained taking advantage of the reducing properties
of chitosan salt and carrying out the reduction process at proper temperature or using
radiation which is a subject of patent application no. P 385153 (2008) [29]. A preliminary
characteristic which allows us to realize the size of formed silver nanoparticles was made
on the basis of the measurements of UV absorption (PERKIN ELMER) and dynamic light
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80
0
2
4
6
8
10
12
14
16
18
20

S
i
l
v
e
r

i
o
n

c
o
n
c
e
n
t
r
a
t
i
o
n

[
m
g
/
d
m
3
]
Time [h]
chitosan acetate, salt
chitosan hydrogel, granules
0 20 40 60 80 100
0
10
20
30
40
50
60
70
80
90
100
110
120
130

S
i
l
v
e
r

i
o
n

c
o
n
c
e
n
t
r
a
t
i
o
n

[
m
g
/
d
m
3
]
chitosan acetate, salt
chitosan hydrogel, granules
Time [h]
Figure 1. Adsorption of Ag
+
from different
forms of chitosan; silver nitrate 10 mg/dm
3
.
Figure 2. Adsorption of Ag
+
from different
forms of chitosan; silver nitrate 50 mg/dm
3
Progress on Chemistry and Application of Chitin and Its ..., Volume XV, 2010 67
Synthesis of Silver Nanoparticles in a Chitosan Solution
dispersion (Malvern Instruments Zetasizer Nano ZS). UV spectra for selected samples are
shown in Figure 3.
The size of silver nanoparticles can be controlled not only by chitosan parameters and the
concentration of silver nitrate, but also conditions in which the reduction process is carried
out. The obtained silver nanoparticles of diameters from 2 m to 50 nm were stable in water
solutions. Size distributions of the obtained particles are given in Table 1.
400 600 800
0
1


(0)
(Chitosan)
(1)
/nm
A
b
s
o
r
b
a
n
c
e
(3)
Figure 3. UV spectra of chitosan acetate solutions containing silver nanoparticles; 0 chelating
process at the temperature 60C in 24 h, 1 chetate compounds of chitosan with silver exposed
to radiation of 10 kGy, 3 chetate compounds of chitosan with silver exposed to radiation of
50 kGy.
Table 1. Size distribution of silver nanoparticles.
Method of
nanoparticle formation
Particle size, nm
Peak analysis by
number
Peak analysis by
intensity
Peak analysis by
volume
0 60 C, 24 h
57.8
2528
97.4
2493
2818.5
95.7
4.3
-
-
-
-
-
1.4
98.6
-
-
-
-
-
100
1 radiation of
10 kGy
284.2
933.4
263.7
967.2
323.1
991.7
97.0
3
-
-
-
-
-
83
17
-
-
-
-
-
50
50
3 radiation of
10 kGy
6.3
9.1
89.4
8.5
100
-
-
-
-
5
95
17
-
-
-
100
Progress on Chemistry and Application of Chitin and Its ..., Volume XV, 2010 68
Z. Modrzejewska, R. Zarzycki, J. Sielski
An optical illustration of the obtained silver nanoparticles is shown in the photo-
graphs taken under an electron scanning microscope FEI Quanta 200 F. They show distinct,
evenly distributed silver particles of the size of several nanometers (Figure 5) which is in
agreement with the results obtained by the method of dynamic light dispersion. Nanopar-
ticles form clusters of several micrometer dimensions with increased intensity (Figure 6).
It is also possible to form structures of the size of several hundred nanometers (Figure 7).
Silver nanoparticles can also be formed into balls (Figure 8). These structures are particu-
larly interesting. Authenticity of silver present in these spherical structures is confrmed by
an elemental analysis performed using Oxford EDS equipped with detector XMAX 50.
3. Conclusions
Chitosan salt can be an interesting nanosilver stabilizer. The size of silver nano-
particles can be controlled not only by chitosan parameters and the concentration of silver
nitrate, but also conditions in which the reduction process is carried out. The obtained silver
nanoparticles of diameters from 2 m to 50 nm were stable in water solutions.
Researches on this subject are promising and require further systematic studies.
Sizedistribution(s)
1 5 10 50 100 500 1000 5000 10000
Diameter (nm) (x10^2)
50
100
%

i
n

c
l
a
s
s
Sizedistribution(s)
50 100 500 1000
Diameter (nm)
10
20
30
40
%

i
n

c
l
a
s
s
Sizedistribution(s)
5 10 50 100 500
Diameter (nm)
10
20
30
40
%

i
n

c
l
a
s
s
Figure 4. Particle size distribution;
a chelating process at the tempera-
ture 60 C in 24 h, b radiation of
10 kGy, c radiation of 50 kGy.
Progress on Chemistry and Application of Chitin and Its ..., Volume XV, 2010 69
Synthesis of Silver Nanoparticles in a Chitosan Solution
Figure 5. Silver nanoparticles with chitosan salt as a stabilizer.
Figure 6. Areas with increased nanosilver concentration.
Figure 7. Nanoparticles of hundred-nanometer size.
Progress on Chemistry and Application of Chitin and Its ..., Volume XV, 2010 70
Z. Modrzejewska, R. Zarzycki, J. Sielski
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