Radiolytic Synthesis of Ag-

loaded Polystyrene (Ag-PS)

Nanoparticles and Their
Antimicrobial Efficiency Against
Staphylococcus aureus and
Klebsiella pneumoniase

Seong-Dae Oh, Bok-Soo Byun, Seungho Lee,

and Seong-Ho Choi

An Oral Presentation by: Model Grace A. Montes

 Introduction
 Nanoscience
- concerned with the
study of the unique
properties of matter at
its nano level
- exploits these
properties to create novel
structures, devices and
systems for a variety of
different uses

*nylon 6 nanofibers decorated with surface bound Ag nanoparticles

 Nanoparticles

 particles having sizes less than 100nm

 have strikingly different properties due to their small
sizes
 Ability to measure and manipulate matter on the
nanometer level is making possible a new generation of
materials with enhanced mechanical, optical, transport
and magnetic properties.
 From micro-nano level : change in its physical as well as
chemical properties:
-increase in the ratio of the surface area to
volume
-size of the particle
 Ag Nanoparticles

 Potentionally useful for applications such as:

a. catalyst for reduction of aromatic nitro
compounds
b. surface-enhanced agent
c. a chiral-enhanced agent
d. an additive for chiral sparation
e. an antimicrobial agent
 Polymer-metal composites

 extensively studied as organic-inorganic

composites
 found to have synergic characteristics of
inorganic materials and polymers
 have potential applications for optical,
catalytic, and magnetic materials
 Previous Synthesis

 Spherical polymeric
particles (beads) were
coated with fine
particles
(nanoparticles) by
mechanical or chemical
method

Polystyrene beads
Mechanical:
 Stirred mixing or high-
shear milling
Chemical:
 Suspension polymerization
or Emulsion polymerization
then deposition of metal
particles

Ag-bound nanoparticles
So far, studies on
radiation induced
preparation of composite
organic-inorganic
particles are SCARCE.

a nanocomposite
 Previous studies show that:

 Polymer particles can be synthesized through

radiation induced polymerization (radiolytic).

 Metallic nanoparticles can be synthesized (e.g.

Ag, Au, Pd, and Pt) using radiation induced
reduction of metal ions.
 Objectives

 This study aims to synthesize Ag-loaded

polystyrene nanoparticles through radiation and
to determine their efficiency against bacteria
such as Staphylococcus aureus and Klebsiella
pneumoniase.
 Scope and Limitations

 Although the morphology of Ag-loaded

nanoparticles were examined, a comparative
study on the use of radiation for loading or
loading distribution of metal particles over
the surface or within polymer particles has
not been attempted.
 Staphylococcus aureus

-a spherical bacterium,
frequently living on the skin or in
the nose of a person
-can cause a pimples, impetigo,
boils, cellulitis folliculitis,
furuncles, carbuncles, scalded
skin syndrome and abscesses,
pneumonia, meningitis,
osteomyelitis endocarditis, Toxic
shock syndrome (TSS), and
septicemia
 Klebsiella pneumoniase

- a Gram-negative, non-motile, encapsulated, lactose

fermenting, facultative anaerobic, rod shaped
bacterium found in the normal flora of the mouth, skin,
and intestines

-can cause bacterial pneumonia, typically due to

aspiration by alcoholics, though it is more commonly
implicated in hospital-acquired urinary tract and wound
infections, particularly in immunocompromised
individuals
 Instrumentation
Energy Filtered
Transmission Electron
Microscopy
-a technique used in
Transmission electron
microscopy, in which
only electrons of
particular kinetic
energies are used to
form the image or
diffraction pattern.
 Field Emission
Transmission Electron
Microscopy

-is able to cope with both

observation of an electron-
microscopic image of a high
brightness and
microanalysis
 Energy Dispersive
X-ray spectroscopy
(EDXS)

-is a chemical microanalysis

technique performed in
conjunction with a scanning
electron microscope SEM

- features or phases as small as

about 1µm can be analyzed.
 X-ray diffraction (XRD)
analysis

-use of X-rays to study the

atomic and molecular
structure of crystalline
substances such as ceramics,
stone, sediments, and
weathering products on
metals
 Methodology

Chemicals:
 Styrene (99%)
 Silver Nitrate

 Poly (vinyl pyrrolidine) (M=40,000)

- All other chemicals were in reagent grade

and were used w/o further purification
Method I (Two-Step Process)
1.0g poly(vinyl pyrrolidine)(PVP) was dissolved
in 200mL EtOH/H2O mixture

1.0g, 1.1 mol styrene was added

The solution was stirred for 30mins. under

nitrogen atmosphere

sol’n was irradiated by gamma-ray from

Co-60 source for 3hours
AgNO3(0.6g,1.0x10-2 M) solution was prepared
in PS colloids of 184mL.

N2(g) was bubbled through the sol’n for 30mins.

Sol’n was irradiated by gamma-ray

from Co-60 source for 3hrs.
Method II (One-Step Process)
PVP(1.0g), styrene (1.0g, 1.1 mol),AgNO3
(0.6g, 6.5 mmol) were dissolved in 100mL
of EtOH/ water mixture (60/40 vol%)

Solution was stirred for 30mins under

nitrogen atmosphere

Sol’n was then irradiated by gamma-ray

from Co-60 source for 3hrs.
Antimicrobial Efficiency Test

Ag-PS nanoparticles were coated on a cloth.

The Ag-PS coated cloth of 0.4g was immersed

in the bacteria sol’n of 0.2mL

Bacteria sol’n was stored for 18hrs at 37◦C

Antimicrobial efficiency was determined by:

% efficiency= Mb-Ma x 100
Mb
•Where Ma and Mb are the conc. of the bacteria before and after the
incubation
“After-washing” antimicrobial Efficiency

Ag-PS-coated cloth was washed at 40+30C, and

dried at room temperature
Characterization of Ag-PS Nanoparticles

Ag-PS nanoparticles were analyzed using

EF-TEM, FE-TEM, and XRD
 Results and Discussion

 Radiolytic Polymerization of styrene was

carried out using a gamma-irradiation in the
presence of PVP in EtOH/H2O.
Advantages:
a. the polymer (PS) is pure
b. free from end fragments of the
initiator
Table I. Effects of Solvent Composition on the Size of
Polystyrene Particles Produced by Radiation-Induced
Polymerization
Vol% EtOH Vol%H2O Particle Diameter (nm) Note

100 500 - -
80 20 - figure1(1)
60 40 257-260 figure1(b)
50 50 98-253 figure1(c)
40 60 80-106 figure1(d)
20 80 73-98 figure1(e)
0 100 60-80 figure1(f)
 Effects of Solvent Composition on the Size of Polystyrene Particles
Produced by Radiation-Induced Polymerization

300

250

200

150 particle
diameter(nm)
100

50

0
60% 50% 40% 20% 0%
 Table1 displays the size of PS particles
obtained with different solvent
compositions as medium of the reaction

 The size of PS particles gradually

decreases as the vol% of water
increases.

 As the vol% of water increases, the

solubility of the PS decreases and thus
the particle precipitates at the early
stages aryl nucleation of smaller
particles.
 PS particles prepared in water are
gelated, hence size could not be
determined

 PS particles formed with 80/20

EtOH/H2O are also partially gelled

 Majority of the PS particles are

aggregated.
-interactions between the PVP’s on
the surface of the PS particles and the
radicals generated through gamma-
irradiation
Table II. Effects of Irradiation Dose on the Size of
Polystyrene Particles Produced by Radiolytic
Polymerization of Ag Ions
Total Irradiation Particle Diameter Note Note
Dose (kGy) (nm)

5 230-260 fig.2(a) fig.2(a-1)

10 250-300 fig.2(b) fig.2(b-1)
15 380-454 fig.2(c) fig.2(c-1)
Effects of Irradiation Dose on the Size of Polystyrene Particles
Produced by Radiolytic Polymerization of Ag Ions

450
400
350
300
250
particle
200
diameter (nm)
150
100
50
0
5 10 30
 Table II shows the effect of
irradiation dose on the size of the PS
particles produced by method I at
room temperature in 60/40
EtOH/H2O

 Average particle size of the PS

particles gradually increases as the
irradiation dose increases from 5 to
30 kGy.
 Fig. 2(a-1), 2(b-1), and 2(c-1) shows the
FE-TEM images of Ag-PS particles
produced by radiolytic reduction of Ag
ions in the presence of PS particles

 Fig.2(a-1),2(b-1), and 2(c-1) reveal that

Ag nanoparticle are of 10-20nm sized

 Ag nanoparticle are loaded on the

surface of the spherical beads of PS
particles
 Fig.3 shows the energy dispersive
X-ray spectroscopy (EDXS) spectra
and XRD pattern of the Ag-PS
composite particles prepared with
30kGy irradiation.

 These results confirm the presence of

Ag nanoparticles on the surface of the
particles.
 PS particles synthesized by Method II
are somewhat uniform in size
(around 250nm).

 Ag particles were found within PS

particles (not just on the surface of
the particles).

 Average size of Ag particles are of

12-25nm.
Table III. “After-washing” Antimicrobial Efficiency of Ag-PS
Particles shown in Figure 4 against Staphylococcus
aureus (ATCC 6538) and Klebsiella pneumoniase (ATCC
4352)
No. of Antimicrobial Efficiency(%) Against
Wasing Cycles ATCC 6538 ATCC 4352

20 99.9 99.9
50 99.9 99.9
70 99.9 99.9
100 99.9 99.9
 The antimicrobial Efficiency of the
cloth coated with the PS-Ag
nanoparticles were 99.9% after 20
cycles of washing

 It remained constant at the same level

even after 100 washing cycles.
 Conclusion
 The two-step method in synthesizing
Ag-PS nanoparticles results in loading
of Ag particles on the surface of the PS
particles.

 The one-step method distributes the

Ag particles within the PS particles.
 When coated on a cloth, the PS-Ag
nanoparticles produced by the one-
step method showed an excellent
antimicrobial activity.

 A cloth coated with those particles

shows a high antimicrobial efficiency
of 99.9% against two bacteria,
Staphylococcus aureus and Klebsiella
pneumoniase.
Recommendation
 a comparative study on the use of
radiation for loading or loading
distribution of metal particles over
the surface or within polymer
particles is recommended
 References (14) K. Torigoe, H. Remita, P. Beaunier, and J. Belloni, 
Radiat.Phys. Chem., 64, 215 (2002).
(15) A.Sárkány, Z. Papp, I.Sajó, and Z. Schay,
Solid State Ionics,176, 209 (2005).
(1) I. B. Jang, J. H. Sung, H. J. Choi, and I. Chin, 
(16) S. Seino, T. Kinoshita, Y. Otome, T. Maki, T. Nakagawa, K.
Synthetic Metals, 157, 9 (2005).
Okitsu, Y. Mizukoshi, T. Nakayama, T. Sekino, K. Niihara,S.­D. Oh
(2) Z. Huang, F. Tang, and L. Zhang, 
et al.290Macromol. Res., Vol. 15, No. 4, 2007
Thin Solid Films, 471, 105(2005).
and T. A. Yamamoto, 
(3) A. Pich, J. Hain, Y. Prots, and H.­J. Adlera, 
Scripta Materialia, 51, 467 (2004).
Polymer, 46,7931 (2005).
(17) N. Pradhan, A. Pal, and T. Pal, 
(4) T. Endo, T. Yoshimura, and K. Esumi, 
Colloid Surface A, 196, 247
J. Colloid Interf. Sci.,286, 602 (2005).
(2002).
(5) W.­J. Liu, W.­D. He, Y.­M. Wang, D. Wang, and Z.­C.
(18) S.­H. Choi and H. G. Park, 
Zhang, 
Appl. Surf. Sci., 243, 76 (2005).
Polymer, 46, 8366 (2005).
(19) S. Sugawa, K. Sayama, K. Okabe, and H. Arakawa, 
(6) H. Nathani and R. D. K. Misra, 
Energ.
Mater. Sci. Eng. B, 113, 228(2004).
Convers. Manage., 36, 665 (1995).
(7) Z. L. Liua, Z. H.Dinga, K.L. Yaoa, J.Taoa, G.H. Dua, Q.H.
(20) T. Li, H. G. Park, H.­S. Lee, and S.­H. Choi, Nano Tech., 15,S660 (2004).
Lua, X. Wanga, F. L. Gongb, and X. Chenc, 
(21) S.­H. Choi, S. H. Lee, Y. M. Hwang, K. P. Lee, and H. D.
J. Magn. Magn.
Kang, 
Mater., 265, 98 (2003).
Radiat. Phys. Chem., 67,517 (2003).
(8) T. Iwasaki, M. Satoh, and T. Ito, 
(22) S.­H. Choi, H. J. Noh, and K. P. Lee, 
J. Mater. Process. Technol.,146, 330 (2004).
Bull. Korean Chem.
(9) B. S. Lee, S. Mahajan, and K. D. Janda,
Soc., 26, 1549 (2005).
Tetrahedron, 61,3081 (2005).
(23) S.­D. Oh, S. Lee, S.­H. Choi, I.­S. Lee, Y.­M. Lee, J.­H.
(10) J.­W. Shim, J.­W. Kim, S.­H. Han, I.­S. Chang, H.­K. Kim,
Chun, and H.­J. Park,
H.­H. Kang, O.­S. Lee, and K.­D. Suh,
Colloid Surface A, 275, 228 (2006).
Colloid. Surface A,207, 105 (2002).
(24) S.­D. Oh, B.­S. Byun, S. Lee, and S.­H. Choi, 
(11) C.­S. Chern and T.­C. Yu, 
Macromol.
Polymer, 46, 1899 (2005).
Res., 14, 194 (2006).
(12) Y. Wang and C. Pan, 
(25) http//www.fiti.re.kr.
Eur. Polym. J., 37, 699 (2001).
(26) S.­H. Choi, K. P. Lee, and S. B. Park, 
(13) S.­H. Choi, K. P. Lee, and H. D. Kang, 
Stud. Surf. Sci. Catal.,146, 93 (2003).
J. Appl. Polym. Sci.,88, 1153 (2003).