Fibers and Polymers 2015, Vol.16, No.

7, 1519-1526 ISSN 1229-9197 (print version)

DOI 10.1007/s12221-015-5245-6 ISSN 1875-0052 (electronic version)

Preparation of Antibacterial Cotton Fabric Using Chitosan-silver Nanoparticles

Dooa Arif, Muhammad Bilal Khan Niazi1*, Noaman Ul-Haq1, Muhammad Nabeel Anwar, and
Emran Hashmi2
Department of Biomedical Engineering and Sciences, School of Mechanical and Manufacturing Engineering,
National University of Sciences and Technology, Islamabad 44000, Pakistan
1
Department of Chemical Engineering, School of Chemical and Materials Engineering, National University of Sciences and
Technology, Islamabad 44000, Pakistan
2
Institute of Environmental Science and Engineering, School of Civil and Environmental Engineering National University of
Sciences and Technology, Islamabad 44000, Pakistan
(Received April 1, 2015; Revised June 9, 2015; Accepted June 11, 2015)

Abstract: The aim of this study was to prepare antibacterial cotton fabric using chitosan-silver nanoparticles (CS-AgNPs).
CS-AgNPs were used as finishing agent for 100 % pure cotton fabric. AgNPs were prepared by Turkevich method. CS-
AgNPs were synthesized by mixing chitosan solution with silver nanoparticles. Fourier transform infrared (FTIR)
spectrometer technique supported the formulation of CS-AgNPs. Cs-AgNPs crystalline peaks were in perfect agreement with
JCPDS card no. 89-3722. Two gram negative bacteria Escherichia coli, Pseudomonas aeruginosa and two gram positive
bacteria Bacillus cereus, Staphylococcus aureus were used to test the bacterial efficacy of synthesized AgNPs and CS-
AgNPs. Scanning electron micrograph of cotton fabric revealed the presence of CS-AgNPs on the surface of cotton fabric.
The presence of small amount of silver nanoparticles in the composite was enough to enhance antibacterial activity
significantly compared to pure chitosan.
Keywords: Chitosan, Antibacterial, Silver nanoparticles, Cotton, Biopolymer

Introduction biodegradability, non-toxicity, cationic nature, and antimicrobial

activity [6]. In agriculture many of the properties served as a
Textile substrates, especially of natural origin, are splendid major elicitor of plant defense mechanisms, as a flocculating
source for the suitable growth of microbes as they contribute agent in wastewater treatment, plus an additional additive in
the conditions like moisture, temperature, oxygen, and food and beverage industry. Currently these agents are as
nutrients that are necessary for their growth [1]. well being utilized in the cosmetic and pharmaceutical
Cellulosic fabrics, i.e. linen, cotton and viscose are extensively industries [7,8]. Chitosan could be an exemplary material for
adopted in the field of clothing. There arenumerous reasons textile industry enduring eco-friendly processes [9].
for their use such as as renewability, eco-friendly, hydrophilic Similarly silver, in oxidation states (AgO, Ag+, Ag2+, and
properties, air permeability and comfortability [2]. However, Ag3+), is commonly used as an antimicrobial agent against
cellulose-based textiles are carriers of micro-organisms and numerous bacterial strains and micro-organisms [10]. AgNPs
more vulnerable to microbial attack than synthetic textiles. are smaller than 100 nm consisting of 20-15,000 silver atoms.
Due to their properties of porosity, hydrophilic nature along Particularly at nanoscale, silver shows incredible physical,
with the ability to maintain water, oxygen, and nutrients, they chemical and biological properties, and antibacterial activity
promote bacterial growth, body odor, and also loss performance [11]. It is commonly used as antibacterial agent due to low
properties [3]. Recently, numbers of antimicrobial-finished toxicity toward mammalian cells [12,13].
textiles have been acknowledged for clinical perspective to Work has been done on the use of silver nanoparticles as
obstruct the transmission of pathogens. These products antimicrobial textile finish. Zhang and Wu showed antibacterial
consist of antimicrobial agents such as silver, quaternary effects of cotton fabric impregnated with colloidal silver
ammonium chloride, and chitosan, and showed anti-bacterial synthesized in one step method using AgNO3 against S.
activity against a wide range of microorganisms [4]. aureus and E. coli [14]. El-Rafie extracted AgNPs from
The steady exploration of antimicrobial agents has led to biomass filtrate of fungus F. solani, particle size 3-8 nm with
recognition of antimicrobial biomaterials that are devised effective antimicrobial activity even if they were used in
from the polymer or are the composites of these polymers. very minute amount of 54 ppm towards the development of
Chitosan is one such poly-cationic biopolymer with high hybrid [15].
antimicrobial activity. Chitin is generally found in shells of Nowadays, efforts have been laid upon hybrid of metallic
marine crustaceans and cell wall of fungi [5]. Chitosan is a salts and biopolymers on the basis of polymer characteristics
natural biopolymer and possess distinctive properties such as comprising of metal ion properties. Chitosan occupy high
chelating strength for numerous metal ions. This chelation
*Corresponding author: [Link]@[Link] enhance the positive charge density of chitosan and increase

1519
1520 Fibers and Polymers 2015, Vol.16, No.7 Dooa Arif et al.

adsorption of polycation onto the negatively charged cell solution with sodium citrate using the standard method
surface erupting improved growth inhibition [16]. The described by Turkevich [23]. Aqueous solution of silver
nanoparticles in combination with chitosan are potential nitrate (1.0 mmol l-1, 500 ml) was heated and stirred gently
candidate in biomedical applications due to advantages of with a magnetic stirrer up to 90 oC. Then preheated sodium
biodegradability, antibacterial and amazing chelating properties. citrate solution (5.0 ml of 0.3 mol l-1) at 90 oC was added to
Numerous work has been done using chitosan with different the above solution. Appearance of yellow color indicates the
nanoparticles, such as Farouk et al. combined the ZnO presence of AgNPs. Further AgNPs were dried using rotary
nanoparticles with different molecular weights of chitosan evaporator and oven drying. After drying fine black particles
showing effective antibacterial properties [17]. Elhady et al. were obtained. The AgNPs were dispersed in distilled water.
used different concentrations of ZnO nanoparticles with The prepared AgNPs solution (1 ml) was added into the
chitosan at different temperatures that induced antibacterial chitosan solution (1 % w/v in 1 % acetic acid) and stirred for
and UV protection in finished cotton fabrics [18]. Silver and 2 hours to obtained CS-AgNPs [24].
chitosan are both antibacterial agents, so CS-AgNPs composite
material has more antibacterial effect [19]. Wazed et al. Preparation of Chitosan-silver Nanoparticles (CS-AgNPs)
prepared chitosan loaded AgNPs for polyester fabric in low Treated Cotton Fabric
concentration. The method used was ionic gelation [20]. Fine cotton fabric was immersed in an aqueous solution.
Another research carried out by Youbo et al. formulated An aqueous solution contains CS-AgNPs along with penetrant
chitosan AgNPs composite as an antimicrobial finish for Keirlon XCJ (mixture of non-ionic surfactants) and an
tencel/non-woven cotton fabric and checked their activity antifoaming agent MONOELAST NT (Non-ionic functional
against [Link] and S. aureus [21]. Thomas et al. have attached poly siloxane). Buffer INVATEX AC (Acidic components
chitosan with cotton using coupling reaction induced covalent based) was added to maintain the pH of the solution.
attachment and then AgNPs were integrated into chitosan Afterwards, passed through padding mangle with 70 % wet
layer by loading Ag(I) ions accompanied by citrate and pick up. The padded samples were dried and cured at 150 oC
investigated their activity with [Link] [22]. for 2 min.
Keeping this background, combination of chitosan and
AgNPs as an effective antibacterial textile finish was prepared. Characterizations of AgNPs and CS-AgNPs
AgNPs was prepared by using turkevich method and were X-ray Diffraction (XRD)
added to chitosan solution in order to prepare textile finish. XRD analysis of AgNPs was achieved using XRD model
The efficiency of the CS-AgNPs was tested on different theta-theta STOE (Germany).
strains of gram-positive and gram-negative bacteria. Textile Ultraviolet Visible Spectroscopy (UV-Vis)
has been coated with this finish to improve its resistant to UV-vis shows the absorbance spectra of silver nanoparticles.
antibacterial attack. The method applied was more convenient UV-vis spectrum of CS-AgNPs was carried out on UV-2800
and rapid as compared to studies mentioned formerly. The BMS Scientific Technical Corporation (PVT) Ltd.
amount of silver nanoparticles was very minute still it Fourier Transform Infrared Spectroscopy
showed the significant effect. This study focused on utilizing FTIR analysis were performed to investigate any structural
less use of chemicals to make it eco-friendly and economical. changes in the CS-AgNPs [24]. The infrared spectra were
recorded on Perkin Elmer, spectrum 100 FTIR spectro-
Experimental photometer.
Scanning Electron Microscopy (SEM)
Materials The visual images of CS-AgNPs were produced by scanning
100 % pure Cotton fiber was provided by Kohinoor textile electron microscopy using JEOL JSM-6042 A (Japan). The
mills, Rawalpindi, Pakistan. Chitosan and Acetic Acid (100 %) working distance was of 10 mm which was maintained thereby
was obtained from Sigma-Aldrich (USA). Silver nitrate was using 5 kV acceleration voltage with a variety of various
obtained from Fisher Scientific (UK), sodium citrate from magnifications. The CS-AgNPs fabric was coated by gold
Etiqueta (CE) and nutrient agar from Merck Specialties using sputtering technique before obtaining measurements.
(Mumbai). The test strains Escherichia coli, Pseudomonas
aeruginosa, and Bacillus cereus, Staphylococcus aureus Antibacterial Activity of CS-AgNPs by Disc Diffusion
were taken from clinical isolates. All the chemicals used in Method
this work were of analytical grade and used as received, AATCC Test method 147 (DIN EN ISO 20645-2001) is a
without further purification. qualitative method for the evaluation of bacteriostatic activity
of the antimicrobial material [25]. The antibacterial activities
Synthesis of Silver Nanoparticles and Chitosan-silver of the fabric were tested against strains of Gram-negative
Nanoparticles (CS-AgNPs) microorganisms Escherichia coli, Pseudomonas aeruginosa,
AgNPs were prepared by the reduction of silver nitrate and Gram-positive Bacillus cereus, Staphylococcus aureus.
Antimicrobial Cotton Fabric Using CS-AgNPs Fibers and Polymers 2015, Vol.16, No.7 1521

During qualitative measurements, samples were placed to 10.6) previously heated to 40 oC to provide a liquid ratio of
form a circular zone. The antibacterial activity of AgNPs, 50:1. Subsequently samples were washed off in cold distilled
chitosan and CS-AgNPs was tested using modified agar water and squeezed and dried at room temperature. The
diffusion assay (disc test) [26]. Plate inspections were carried quality of coated samples was assessed after 20 washes.
out to note down the zone of inhibition at 38 oC for 24 hours.
Any prominent zone of inhibition around the samples was Results and Discussion
recorded as an inhibitory effect against microbial species.
The coated fiber samples were tested in order to detect the Characterization of Silver Nanoparticles, Chitosan and
presence or absence of bacterial growth in the contact region. Cs-AgNPs
Two different weave of cotton fabrics were tested against X-ray Diffraction (XRD) of Silver Nanoparticles
microorganisms. The X-ray diffraction patterns of the chitosan, CS-AgNPs
and Silver nanoparticles were shown in Figure 1. The
Antibacterial Evaluation of CS-AgNPs Coated Cotton chitosan showed peak at 10 o and 20 o correspond to a
Fabric Using Quantitative Method hydrated crystalline structure and the peak at 20 o an
Antibacterial testing was carried out by using the shake amorphous structure of chitosan, respectively [27-29].
flask method ASTM E2149. Test was conducted against the In the XRD pattern of CS-AgNPs, the peaks at 10 o and
Gram-positive Staphylococcus aureus and Gram-negative 20 o showed the characteristics peaks of chitosan and the
Escherichia coli. One gram of fabric was placed in a conical peaks obtained at 38 o, 44 o, 64 o and 77 o in CS-AgNPs are
flask with 0.5 ml of bacterial inoculum in 50 ml of sterile referred to Silver nanoparticles [30].
broth. The flask was shaken at 37 oC at 150 rpm for 24 hours In the XRD pattern of Silver nanoparticles, peaks were
at shaking platform. After incubation of 24 hours contact obtained at 2 theta values of 38 o, 44 o, 64 o and 77 o that
time the solution was serially diluted. The diluted solution correspond to the (111), (200), (220) and (311) reflections of
was plated on nutrient agar and incubated for 24 hours at face centered cubic (fcc) silver which are in perfect agreement
37 oC. The reduction of colonies was calculated using the to the JCPDS card no. 89-3722 [30]. Similarly, no spurious
following equation: diffraction was observed to indicate the crystallographic
impurities in the sample. Coincided peaks confirmed the fcc
(B – A)
R% = ---------------- × 100 structure of pure silver metal. The (111) peak was detected in
B
the sample, representing high intense peak of fcc materials.
where A is the number of bacterial colonies survived after The high degree of crystallinity of the AgNPs was revealed
contacting with treated sample and B is the number of by the intensity of peaks. However, the broad diffraction
colonies present in untreated control sample. peaks were expressing the small size of crystals [31].
The average crystallite size of the AgNPs was calculated
Washing Fastness Test from the XRD line broadening using the Scherrer equation
CS-AgNPs coated cotton fabrics were tested after repetitive [32].
washings in AATCC Atlas Launder-O-Meter Standard
0.93λ
Instrument. This is used broadly on laboratory scale for B = --------------
Lcosθ
assessing laundry result. One wash that is carried out in
Launder-O-Meter [ISO-105-CO1: 1989 (E) Standard method] where λ is the wavelength of the incident X-rays (λ=
is equivalent to five home laundry washings. The washing 1.54060 Å), L is the full width at half maximum of the (111)
cycles time span was 30 min and solution utilized was of diffraction, and θ is the angle of diffraction. The diameter of
SDC standard detergent with the concentration of 5 g/l (pH AgNPs is 27 nm.

Figure 1. XRD of chitosan, CS-AgNPS and silver nanoparticles.

1522 Fibers and Polymers 2015, Vol.16, No.7 Dooa Arif et al.

Figure 2. UV-vis spectrum of chitosan, AgNPs and CS-AgNPs.

Figure 3. FTIR of silver nanoparticles, chitosan-silver nanoparticles,
and chitosan.
UV-VIS Spectroscopy of CS-AgNPs
UV-vis spectroscopy was performed to study the structural
characterization of AgNPs present in CS-AgNPs. Surface
plasmon resonances (SPRs) dominated the optical spectra of
the metal nanoparticles thereby transferring to longer
wavelengths with an increment in particle size [33]. UV-vis
spectroscopy is a reliable method for detecting the presence
of one-dimensional metallic nanostructures [34,35]. Furthermore,
the size and shape characteristics rely on the absorbance
pattern of AgNPs [36]. The SPR peaks are inversely
proportional to the symmetry of nanoparticles. As the total
number of SPR peaks decreases, the nanoparticle symmetry
rises [37]. The UV-vis spectrum of CS-AgNPs was demonstrated
in Figure 2. A single peak aroused at 434 nm in the spectrum
of CS-AgNPs due to an excitation in the surface plasmon
vibrations of silver atoms [24].
FTIR of Silver, Chitosan and Chitosan-silver Nanoparticle
Composite
The characteristic peaks of chitosan were observed at
3398 cm-1 (OH stretch overlapped with NH stretch), at 2877 cm
(shows the presence of sp3 C-H stretch), at 1601 cm-1 (shows
the presence of C=O vibration of amide group), at 1384 cm-1
(shows CH stretching), and at 1091 cm-1 (shows C-O-C
stretch) and 667 cm-1(shows stretching vibration of CH bend
out of plane) [22,38].
The FTIR analysis of silver nanoparticles showed peaks at
3436 cm-1 (O-H Stretching vibrations), 2922 cm-1 (C-H
Figure 4. SEM micrographs of (a) untreated sample and (b) treated.
stretching vibrations), 1590 cm-1 (N-H bending of primary
amines) [30] 1082 cm-1 (C-N stretching vibrations of all
amines), 909 cm-1 and 850 cm-1 (C-H bending vibrations out supporting the formation of CS-AgNPs.
of plane) (Figure 3). SEM Analysis of CS-AgNPs Coated Cotton Fabric
The characteristic peaks of CS-AgNPs were present at SEM analysis was executed to explore changes in the
3212 cm-1 (O-H stretching vibration), 1638 cm-1 (C=O stretch, topography of fabric after treatment process compared to
N-H bending for primary amides), 1037cm-1 (C-N stretching untreated blank sample. SEM micrograph reveals the presence
band for all amines), 722.56cm-1 (C-H bending out of plane). of CS-AgNPs on the cotton fabric, as it is evident from
All these peaks were available in the composite (Figure 3), Figure 4(a), (b). The untreated sample showed plain smooth
Antimicrobial Cotton Fabric Using CS-AgNPs Fibers and Polymers 2015, Vol.16, No.7 1523

surface, whereas treated sample had particles dispersed on The elemental composition of CS-AgNPs coated fabric
its surface. It was mentioned in the literature that untreated was studied by energy dispersive analysis of X-rays (EDAX).
cotton fabric showed plain, smooth and uniform surfaces as Figure 5 depicts the EDAX analysis from a selected area.
compared to the treated cotton [39]. The EDAX analysis confirmed that the CS-AgNPs contained
about 0.10 wt % Ag, 40.03 wt % carbon, and 59.87 wt %
oxygen (Table 1).

Antibacterial Activity
As the purpose of this Antibacterial textile finish was to
resist the growth of bacteria on to the fabric. First of all, the
antibacterial efficacy of AgNPs, chitosan and CS-AgNPs
were examined separately using discs. Figure 6(a), (b)

Table 2. Antimicrobial activity of chitosan, silver and chitosan-

silver nanoparticles against gram negative bacteria, [Link] and
gram positive bacteria, B. cereus
Microorganism Antimicrobial Zone of inhibition
Figure 5. EDAX profile of chitosan-silver nanoparticles (CS- Escherichia coli Silver nanoparticles 15 mm
AgNPs). (gram negative)
Escherichia coli Chitosan 14 mm
Table 1. Elemental composition of chitosan-silver nanoparticles Escherichia coli Silver and chitosan 21 mm
composite material
Element Weight (%)
Bacillus cereus Silver nanoparticles 17 mm
CK 40.03 (gram positive)
OK 59.87 Bacillus cereus Chitosan 11 mm
Ag L 0.10 Bacillus cereus Silver and chitosan 20 mm
Total 100.0 composite material

Figure 6. Zone of inhibition of antimicrobial agents against gram negative and gram positive bacteria (a) Escherichia coli (1) silver
nanoparticles (2) chitosan (3) chitosan-silver nanoparticles, (b) Bacillus cereus (1) silver nanoparticles (2) chitosan (3) chitosan-silver
nanoparticles, (c) Bacillus cereus, (d) Escherichia coli, (e) Pseudomonas aeruginosa, and (f) Staphylococcus aureus.
1524 Fibers and Polymers 2015, Vol.16, No.7 Dooa Arif et al.

showed inhibition zones of AgNPs. Chitosan and CS-AgNPs inhibition.

against [Link] and B. cereus prior on the cotton fabrics. This Figure 6(c), (d), (e), and (f) explained the antibacterial
was done to detect their antibacterial potency. Table 2 performance of coated cotton fabrics of two different weaves
presented the diameter measurements of the zone of against gram positive bacteria Bacillus cereus, Staphylococcus
aureus and gram negative Escherichia coli, Pseudomonas
Table 3. Zone inhibition diameters (mm) of antimicrobial cotton aeruginosa bacteria. Table 3 presented the diameter mea-
fabric against gram negative and gram positive bacteria surements of the zone of inhibition. The quantitative method
Coated fabric Coated fabric showed bacterial reduction in percentage. The antibacterial
Microorganisms activities of cotton fabric containing CS-AgNPs showed
(duck weave) (percale)
high activity with 99.01% bacterial reduction against S.
Bacillus cereus 25 mm 20 mm
aureus and 98.78 % against [Link] (Table 4). The antibacterial
Staphylococcus aureus 29 mm 27 mm efficiency was found to be 98.95 % against S. aureus and
Pseudomonas aeruginosa 30 mm 30 mm 85.82 % against [Link] after 20 washes (Table 5). The
Escherichia coli 24 mm 26 mm antibacterial activity shown by CS-AgNPs coated cotton

Table 4. Antibacterial efficiency of the control and CS-AgNPs coated cotton fabric
Survival cells (CFU/ml) Antibacterial reduction
Fabric sample Test organism
Control fabric Treated fabric (%)
-2 -3
Cotton fabric finished with S. aureus 458×10 45×10 99.01 %
CS-AgNPs E. coli 247×10 -2
30×10 -2
98.78 %

Table 5. Antibacterial efficiency of the control and CS-AgNPs coated cotton fabric after 20 washes
Survival cells (CFU/ml) Antibacterial reduction
Fabric sample Test organism
Control fabric Treated fabric (%)
-2 -3
Cotton fabric finished with S. aureus 458×10 48×10 98.95 %
CS-AgNPs E. coli 247×10 -2
35×10 -2
85.82 %

Table 6. Physical properties of uncoated and CS-AgNPs coated cotton fabric

Physical properties Uncoated fabric Fabric coated with CS-AgNPs Test method
Type of weave Duck weave Duck weave
Fiber content 100 % cotton 100 % cotton AATCC 20/20A
Tensile strength Warp: 72.8 lbs Warp: 75.4 lbs ASTM D 5034
Weft: 57.9 lbs Weft: 47.5 lbs
Tear strength Warp: 62 N Warp: 62 N ASTM D1424
Weft: 23.6 N Weft: 23.6 N
Resistance to abrasion B/W 15000 REV - Not broken B/W 15000 REV - Not broken ISO 12947
Resistance to pilling B/W 125 REV: 4.5 B/W 125 REV: 4.5 ASTM D 4970
B/W 500 REV: 4.0 B/W 500 REV: 4.0
B/W 1000 REV: 3.5 B/W 1000 REV: 3.5
B/W 2000 REV: 3.0 B/W 2000 REV: 3.0
Absorbency 3-4 sec 3-4 sec Drop method
Whiteness 73.47 73.54 AATCC 110
o o
Crease recovery angle Warp: 66 Warp: 62 AATCC 66
o o
Weft: 67 Weft: 67
Color fastness to rubbing Dry: 4.5 Dry: 4.5 AATCC 08
Wet: 4.5 Wet: 4.5
Dimensional stability Warp: -2.1 % Warp: -1.5 % BE EN ISO 6330: 2012
Top loading machine Weft: -1.5 % Weft: -1.0 %
Color fastness to washing Shade change: 4.5 Shade change: 4.5 ISO 105 CO1
(Launder-o- meter) Staining: 4.5 Staining: 4.5
Antimicrobial Cotton Fabric Using CS-AgNPs Fibers and Polymers 2015, Vol.16, No.7 1525

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