Functional and Antimicrobial Properties of herbal nanocomposites from Piper Betle [611074]

Journal of Coatings Technology and Research
Functional and Antimicrobial Properties of herbal nanocomposites from Piper Betle
Plant Leaves for enhanced Cotton Fabrics
–Manuscript Draft–

Manuscript Number: JCTR-D-19-00368
Full Title: Functional and Antimicrobial Properties of herbal nanocomposites from Piper Betle
Plant Leaves for enhanced Cotton Fabrics
Article Type: Original Research
Keywords: Piper Betle; Herbal nanoparticles; Nanocomposite; UV-protection; Chitosan;
Antibacterial activity.
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Functional and Antimicrobial Properties of herbal nanocomposites from Piper
Betle Plant Leaves for enhanced Cotton Fabric s
Subramani Karthik ,1 Kolathupalayam Shanmugam Balu ,2 Rangaraj Suriyaprabha ,2 Vinoth
Murugan,2 Surendhiran Srinivasan,2 Oscar Komla Awitor ,3 Chiristophe Massard ,3
Venkatachalam Rajendran ,2,3,*
1Department of Biotechnology , Dr. N G P Arts and Science College,
Dr. N. G. P. Kalapatti Road, Coimbatore -641048
2Centre for Nanoscience and Technology, K.S. Rangasamy College of Technology,
Tiruchengode –637215, Tamil Nadu, India.
3Centre for Nanoscience and Technology, Dr. N G P Arts and Science College,
Dr. N. G. P. Kalapatti Road, Coimbatore -641048
4Laboratory C -Biosenss EA 4676, Clermont -Ferand University, Université ď Auvergne,
Clermont -Ferrand, France.
Corresponding author:
*Centre for Nano Science and Technology, K. S. Rangasamy College of Technology,
Tiruchengode -637215, Tamil Nadu, India
*Centre for Nanoscie nce and Technology, Dr. N G P Arts and Science College,
Dr. N. G. P. Kalapatti Road, Coimbatore -641048
Tel.: +91 99941 37373
E-mail address : [anonimizat] , [anonimizat] (Venkatachalam
Rajendran). Manuscript
Click here to download Manuscript Manuscript.docx
Click here to view linked References
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ABSTRACT
In the current times , application of nanoparticles in textile industry has become
increasing ly high due to the possibility of having anticipated properties , such as captivating
colors, superior stability, antibacterial activity and high-end UV-protection to the fabrics. In this
study, natural herbal nanoparticles of different sizes are prepared from room shade dried leaves
of Piper betle employing ball milling technique . Going forward, s tructural, morphological, UV –
protective and anti bacterial properties of herbal nanocomposites coated on fabrics are thoroughly
analyzed and interrelated with un -coated fabrics. Herbal nanoparticles were amalgamated with
chitosan to make nanocomposites and are coated on cotton fabrics with the help of pad-dry cure
method. The analysis was done to study physical properties of the coated fabrics such as air
permeability, crease recovery, tensile streng th, tearing streng th, thickness and bursting strength ,
explicitly shows that coated fabrics have better functional properties as compared to un-coated
fabrics. Along with the same lines , herbal nanoparticles reflected good anti bacterial and UV –
absorption propert ies as compared to un-coated and chitosan coated fabrics. Comprehension of
functional properties revealed that herbal nanoparticles coated fabrics highlights their potential
applications of Piper betle nanoparticles in protective clothing.
Keywords: Piper Betle ; Herbal nanoparticles; Nanocomposite; UV-protection; Chitosan ;
Antibacterial activity .

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1. Introduction
Nanoparticles play a critical yet unique role in an array of fields owing to its unmatched
properties and applications [1]. Textile is an impeccable growth medium for microbial growth as
it is rich in organic compounds that are appropriate for biofilm formation and sweat absorption ,
This, in turn, i s responsible for provid ing moisture conditions and caus ing infection s to human .
Comm only, Staphylococcus aureus (S. aureus ) and Staphylococcus epidermidis (S. epidermidis )
bacteria leads to skin infections like boils, impetigo and cellulitis, and furuncle. In textile,
microorganisms entail to negative effect such as dreadful odor, discoloration and reduce the
texture efficiency [2, 3]. The aforementioned issue is conventionally combated by antibacterial
finishing process with the help of aldehydes, halogens, quaternary ammonium compounds and
amines [4].
Chemical antibacterial finishing creates best control of microbial growth , but it also
accompanies some drawbacks as well, such as toxic, non -biodegradable, non -eco-friendly , and
cost effective, etc. Hence , it is indispensable to change reliable, non -toxic, non -allergic, eco –
friendly materials for textiles with antibacterial finishing. The aforementioned factors are
possible by making use of biological materials particularly with herbal plants. Additionally , the
utilization of plant extracts and bioactive compounds encompasses a range of antibiotic
properties , wh ich is conventionally utilized in therapeutic treatments [5, 6]. Moreover , herbal
molecules are secure as they can easily overcome the resistance generated by the pathogenic
microbes . This is possible because they are in collective form that contains more than one
molecule in the protoplasm of the plant cell [7].
In this study, we have selected one of the herbal plants , i.e., Piper Betle that belongs to
Piperaceae family. It has good antibacterial activity against S. aureus , Proteus vulgaris,
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Escherichia coli (E. coli) , and Pseudomonas aeruginosa, etc. Typically, t he antibacterial activity
occurs because of the presence of sterol in leaf extract of Piper betle . The mode of action is
through surface interaction with bacterial cell wall and cell membrane, and alters the primary
structure of cell wall leads to generate pore and worsen the both gram positive and negative
bacteria [8-10]. The Piper betle encompasses phenol components , like ally p yrocatechol, cavicol,
cavibetol, carvacrol, eugenol, and safrole [11, 12 ]. Ally pyrocatechol, and cavibetol incorporates
radio protective activity with the process inhibition through the radiation induced lipid
peroxidation mechanism [13, 14]. When it is used for textile applications , such compounds can
provide better antibacterial activity and UV -protection .
In textiles antibacterial agents coated on fabrics using physic o-chemical agents for
binding. Bio -nanocomposites materials are currently used in material science, nanoscience fields.
Bio-nanocomposites consists polymeric compounds with materials at nano scale [ 15, 16].
Chitosan is a glucosamine biopolymer posse’s immob ilize the preferred biomolecule because its
presence of reactive amino groups, anti microbial, biocompatibility, fabulous film formi ng
nature, non toxicity and [16 -20]. In addition it is positively charged polymer bond with
negatively charged surface, muco adhesive, and wound healing properties attractiv e for
biomedical a pplications [21 -23].
In this current study, Piper betle nanoparticles are produced by using shade dried plant
leaves with the help of ball milling. As prepared Piper betle nanoparticles are preliminarily
observed to take up qualitative studies for s urface morphology, size, crystalinity and purity.
Moving ahead, t he synthesized herbal nanoparticles are blended with chitosan to prepare a
polymeric nanocomposite for coating on fabric. The coate d fabrics are thereafter subjected to
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UV-protection and antibacterial investigation . Also , functional st udies such as strength (Tensile
and Tearing) properties, air permeability , and crease recovery are tested.
2. Materials and Methods
2.1. Plant leaf sample collection
Fresh, healthy and mature Piper betle leaves were cleaned without having any disease
symptoms collected from places in Namakkal, Tamil Nadu, India. Thereafter, these leaves were
thoroughly washed with running tap water , including double distilled (DD) water for several
times to alleviate all the dust particles. Moving ahead , the leaves were then air dried under shade
at room temperature for around 2 weeks.
2.2. Preparation of herbal nanoparticles
The 20 g of shade dried leaves were primarily ground via top down approach with the
help of the mixer grinder to prepare the coarse powder. In order to obtain the fine powder, the
coarse powder was milled for 1 h with the help of 20 mm sized ball ( Zirconia) by making use of
ball mill (PM100; Retsch, Germany) . Along with this , the obtained fine powder was (nearly 7
gm) distributed into three equal parts. The n, these equal fine powder sections were again milled
(10 mm balls: 300 rpm) in a ball mill at distinct milling periods , like 5, 10, and 15 h (hereafter
termed respectively as sample PBNp1, PBNp1, and PBNp3) [24-27].
Fig. 1 schematically represents t he preparation of herbal nanoparticles from Piper betle
plant leaves . Thereafter, t he nanoparticles attained from the above three collected samples
(PBNp1, PBNp2 and PBNp3) were characterized carefully and were further utilized for coating
on fabrics . This was then followed by the analysis of their functional properties.

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Fig. 2. Schematically represents the preparation of herbal nanoparticles from Piper betle plant
leaves .
2.3. Characteris ation
The phase finding of the Piper betle nanop owders was performed with the help of X-ray
diffraction method utilizing a (Philips XRD; X’ PertPro; PANalytical, Netherlands) X pert MPD
powder diffractometer . The powder XRD was obtained in the 2θ range from 10ș to 80ș in a step-
scan mode with 2θ step of 0.02ș . When bombarded with high -energy X -rays for elemental
analysis , the attained leaf nanoparticles were excited.
Further , the synthesis of leaf nanop owders was carefully monitored on a periodic basis in
a UV –visible (UV -vis) spectrophotometer (Cary 8454, Agilent technologies, Singapore)
functioned from the UV to near infrared (NIR) (180 –800 nm) spectral regions at a step size of
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5Å. Thereafter, a volume of 0.1 mL of the sample was diluted in a cuvette in a 2 mL of deionized
water. At a resolution of 1 nm , the UV–vis spectra of the resulting diluents were analyzed as a
function of reaction time and biomaterial dosage. Using dynamic light scattering (DLS)
technique, t he size of the particle s was analysed with the help of a sub-micrometer particle size
analyzer (Nanophox, Sympatec, Clausthal -Zellerfeld, Germany) . The particle size of the all the
obtained herbal nanopowders samples were measured at a scattering angle of 900 and in the
range of 1 -1000 nm. The prepared herbal nano particles was thereafter taken to the scanning
electron microscope equipped with energy -dispersive X -ray analysis (SEM -EDX; JSM 6360;
JEOL, Japan) to determine the morphology, microstructure, and elemental composition of the
collected samples.
2.4. Prepar ation of herbal nanoparticles -chitosan composite
The obtained herbal nanoparticles (1 gm) were mixed in a 100 mL of double distilled
water , including 1% chitosan (from shrimp shells, ≥75% deacetylation, Himedia, India ) was
dissolved in a 1% acetic acid was added . The solution was kept under stirring at a 60 °C till the
time a fine homogenous suspension was formed. Thereafter , the obtained suspension was kept
overnight to allievate all the air bubble . Along with that, the solution was soni cated for around 30
min before making use of it [28].
Now, the bleached and mercerized woven cotton fabric (100%, mass 138.84 g m-2, 116
ends per Inch 84 picks per inch) was pre -treated as a substrate for coating. Then, the cleaned
fabrics were dried in an oven at 50 °C for around 5 min. It is important to note that it is
imperative to add chitosan in order to make the herbal nanoparticle s stable in an aqueous
solution for a long er period of time . A bleached (60 × 60 cm) cotton fabric was then individually
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immersed four times in a homogeneous solution containing chitosan sol and Piper betle -chitosan
solution (nanocomposite finishing). Then, the obtained solution was passed through a padding
mangle at a rate of 35 rpm fo r around 5 min with the help of a DC motor to collect the uniformly
coated fabric . Thereafter, it was dried at 80 °C for 10 min [28, 29 ].
2.5. Ultraviolet protection on coated fabrics
Ultraviolet transmission spectra (Lambda 35; PerkinElmer, USA) with the wavelength
ranging from 280 to 400 nm were used to test the UV blocking properties of the fabric samples.
The total percentage of UV blocking was measured with respect to ASTM D6603 standard [30].
Below is the value of the UV-protection factor (UPF) that was obtained with the help of the
following relation:
UPF =∑ 𝑬(𝝀)×𝑺 (𝝀)×𝜟𝝀𝟒𝟎𝟎
𝟐𝟗𝟎
∑ 𝑬 (𝝀)𝟒𝟎𝟎
𝟐𝟗𝟎 ×𝑺(𝝀)×𝑻(𝝀)×𝜟𝝀 (1)
wherein, S (λ) stands for the solar spectral irradiance (W m-2 nm-1), E (λ) stands for the relative
erythemal spectral effectiveness, and T (λ), stands for the spectral transmittance and Δ (λ), the
wavelength interval .
2.6. Physical properties
Further , the thickness of the coated and un-coated fabric samples was obtained with the
help of fabric thickness tester with respect to ASTM D 5729 -97 guidelines , ranging from 0 to 10
mm. The tensile strength of the un-coated fabrics and coated fabrics was evaluated by making
use of strip method and tensile testing machine (E091; Eureka, India) with respect to ASTM
D5035 -95 standard [31, 32]. Apparently, the testing was auctioned utilizing warp and weft yarns
of the prepared fabric samples. Also, the in crease recovery analyzer (EC -41; Eureka, India) was
used to calculate the crease recovery angle in accordance to AATCC 66 -1998 standard with a
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scale rang ing from 20° to 80° , with an applied creasing load of 300 g for 2 min [33]. The air
permeability values of coated and un-coated fabrics were obtained as per the IS: 11 056-1984 and
DIN 53887 standard with the flow rate of 100 Pa for passing the air perpendicularly utilizing an
air permeability tester (M021A; Premier, India) [34].
2.7. Antibacterial activity
2.7.1. Collection of bacterial cultures and preparation
The cultures of gram -positive S. aureus (ATCC 6538P) and Gram -negative E. coli
(ATCC 9677) were obtained from the renowned National Collection of Industrial
Microorganisms (NCIM), National Chemical Laboratory, Pune, India. Bacterial inoculum was
prepared by carefully inoculating a loopful of test organisms into a nutrient broth and then
incubated at 37 °C for 5 to 8 h till the time a moderate turbidity was developed. Going forward, a
loop of culture was swabbed on the Mueller Hinton agar media.
2.7.2. Agar well diffusion method
The qualitative antibacterial evaluation was performed for E. coli and S. aureus using
agar well diffusion method . Fine well was made in the solidified and inoculated agar medium
(Mueller Hinton agar, Himedia, Mumbai ) utilizing sterile cork borer. The diameter of 7 mm was
maintained on each zone as in all plates. All the wells were loaded with distinct mass
concentrations such as 25, 50 and 100 mg ml-1 of each prepared PBNp1, PBNp2 and PBNp3
herbal nanoparticles . The plates were then incubated at 37 °C for around 24 h. After completion
of the incubation period, the diameters of the inhibition zones were calculated on a milli meter
ruler.

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2.7.3. Agar Disc diffusion method
According to standard size (each pieces with 10 mm size) , the un-coated cotton fabrics
(UC-CF) and polymer/nanocomposites coated (CF -Chi and CF -PBNp -Chi) fabrics were divided
into different samples pieces. Then, the prepared specimen was tenderly pressed in a transvers e
manner across the inoculums of streaks . This was done to make sure the close contact s with agar
surface. In addition to this, the plates were incubated for 18 to 24 h at 37 °C. The aforementioned
process was followed to perform the antibacterial assessments of UC -CF, CF-Chi and CF -PBNp –
Chi fabrics against E. coli and S. aureus.
2.8. Percentage reduction test
In this test, the un-coated (UC -CF) and polymer/nanocomposites coated fabrics (CF -Chi
and CF -PBNp-Chi) were cut into round shaped pieces (10 mm) as recommended by the
American Association of Textile Chemists and Colonists (AATCC 100). The obtained test
samples were then properly soaked into sterile AATCC bacteriostasis broth . After that, a loop
full of tes t organisms was inoculated . The obtained test samples were incubated at 37 °C for
around 18-24 h. Then, serial dilutions from 10-1 to 10-7 were made for all the prepared samples.
By using spread plate method , 0.1 ml of sample from each dilution w as plated on the sterile
AATCC b acteriostasis agar plates , and then, incubated at 37 °C for 24 h. On completion of
incubation period , final concentration of colonies (B) in control and the obtained test samples
were measured using viable cell count method. As the u ntreated fabric (control) contains no
bactericidal activity, the final number of surviving cells will be greater than the initial cell
concentration (A) , which was evaluated through a viable cell count method [28]. The percentage
of bacterial reduction was evaluated with the help of the below mentioned formula :
% Bacterial Reduction= [(A -B)/A] x100 (2)
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wherein, A stands for the initial number of cells and B for the final number of cells.
2.9. Wash durability of coated fabrics
The IS: 687 -1979 standard was utilized to find out the wash durability of CF -Chi and CF –
PBNp -Chi fabrics [35]. This test was performed using a neutral soap at 40±2 °C for around 30
min. The test fabrics were dried out to assess the control for antibacterial activity with the help of
AATCC 100 procedure , which involved up to 15 laundering cycles [36].
3. Results and Discussion
3.1 Characterization of P. betle nanoparticles
The shade -dried Piper betle herbal nanoparticles are prepared by ball milling. The
obtained nanoparticles are milled for a different milling period s (5, 10 and 15h) at 300 rpm .
Then, these nanoparticles are characterized by using different characterization techniques . The
XRD pattern of the prepared nanoparticles namely PBNp1, PBNp2 and PBNp3 is depicted
clearly in Fig. 2A. The obtained results ensure that there are no diffraction peaks at 2 𝜃 values in
the range of 20 –30°. The experimental results explicitly confirm that different parameters of
samples depict amorphous nature with no crystalline peaks .
Fig. 2B shows t he UV -absorption spectra of Piper betle herbal nanoparticles synthesized
from Piper betle plant leaves . In this analysis , all three rotation periods of Piper betle possess the
same UV-absorbance region that ranges from 280 to 284 nm. Such absorbance (280 -282 nm) in
the UV -region allows utilizing the herbal PBNp nanoparticles for UV -protection applications.
Fig. 2C shows t he particle size analysis of PBNp1, PBNp2, and PBNp3 . It is quite evident from
Fig. 2C that the average particle size of PBNp1, PBNp2, and PBNp3 samples is 81, 55, and 26
nm, respectively . These found sizes decreases from 25 to 30 nm as per the rotation .
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The surface morphological properties of PBNps are evident from the SEM images (Fig.
2D). The topography of the SEM image shows clear signs of herbal PBNp nanoparticles that
appear to be agglomerated with cluster like structure. As expected, the elemental analyses
confirm ed the presence of identical elements across all the collected samples (Table 1). In this
study, the high radiation at high energies for O, Na including at low energies fo r Mg, Si, Cl, K
and Ca are seen for PBNp nanoparticle s (Fig. 2D).
The qualitative evaluation of the antibacterial activity of the herbal nanoparticles
(PBNp1, PBNp2 and PBNp3) possesses intriguing observation s, which is quite evident from Fig.
2E. The zone of inhibition at diverse concentrations of different ball milling times (PBNp1,
PBNp2 and PBNp3 ) namely 25, 50 and 100 mg ml-1 of PBNp nanoparticles is shown in Fig. 2E
and later concluded in the Table 1.
The highest formation of zone of inhibition for E. coli (27.24 ± 0.12 ) and S. aureus
(31.20 ± 0.13 ) is observed to be 26 nm for the sample PBNp3 . This indicates higher time period
of milling (15 h) of the plant l eaf powders. At this increased milling time, the size of the
nanoparticles reflected is only 26 nm , which is very low as compared to other milling time. The
smaller particles are observed to possess the formation of maximum zone of inhibition as
compared to the larger particles. The lar ger surface area of smaller nanoparticles aids in seamless
penetration and dena ture of the bacterial cell wall . This , in turn , refrain the DNA replications
[37-39] and thereby leads to formation of wider zone of inhibition than the small surface area of
the larger nanoparticles [24]. The current studies ensure that the surface area of nanoparticles
have an impact on antibacterial property of PBNp s. By carefully experiment ing the
characterization outcomes of all the three collected samples of PBNp, the PBNp3 is opted for
coating process and that’s why it is connoted as PBNp.
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Figure 2. XRD (A), UV-Vis spectrum (B), particle size distribution (C), SEM /EDX (D) and
Antibacterial activity (E) of three different milling period (PBNp1, PBNp2 and
PBNp3 ) PBNp nanoparticles
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Table 1 Antibacterial activity of the Piper betle nanoparticles by agar well diffusion assay

3.2 Characterization of un -coated and coated fabrics
Fig. 3a illustrates t he surface morphological properties and EDX analysis of un-coated
cotton fabrics (UC -CF). On the fab ric surface [Figs. 3b (i and ii)], the SEM image of the chitosan
coated cotton fabrics (CF -Chi) elucidates cluster -type polymer structu res prior to wash. Figs. 3c
(i and ii) reveals th e SEM image and EDX analysis of the CF -PBNps -chi nanocomposites which
are clearly immersed on the fabrics su rface area.
The chitosan polymer is sturdily adhered to the surface of the fabrics even after 5th and
10th washes [Figs. 4a(i) and 5a(i)]. Thereafter, t he wash durability (fastness) of CF -Chi fabric
surface is further ensured with the help of EDX measurements [Figs. 4a (ii) and 5a (ii) ] that is
carried out after 5th and 10th washes. Fig. 4 c elucidates the even dispersion of PBNp -Chi
nanocomposites particles .

Herbal
nanoparticles
Test
organisms Concentrations of Piper betle nanoparticles (Zone
of inhibition (mm))

25 mg ml-1 50 mg ml-1 100 mg ml-1
PBNp1 E. coli 18.52 ± 0.43 21.40 ± 0.12 23.00 ± 0.08
S. aureus 20.08 ± 0.42 23.03 ± 0.28 27.20 ± 0.21

PBNp2 E. coli 19.74 ± 0.02 22.68 ± 0.32 25.56 ± 0.41
S. aureus 22.20 ± 0.23 25.20 ± 0.03 28.38 ± 0.06

PBNp3 E. coli 20.14 ± 0.05 24.50 ± 0.07 27.24 ± 0.12
S. aureus 23.80 ± 0.06 26.07 ± 0.05 31.20 ± 0.13
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Fig. 3. SEM image s and EDX spectra of coated and un -coated fabric before wash
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Fig. 4. SEM image s and EDX spectra of a) CF -chi coat ed fabric and b) CF -PBNp -chi coated
fabric after 5th washing

Moreover, it is also reflected elemental composition analysis on coated fabrics prior to
wash. The presence of PBNp -Chi nanocomposites particles and their wash durability after 5th
and 10th washes are proven through SEM and EDX analysis respectively in Figs. 4b (i and ii) and
5b (i and ii).

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Fig. 5. SEM image s and EDX spectra of a) CF -chi coat ed fabric and b) CF -PBNp -chi coated
fabric after 10th washing

The aforementioned observation clearly shows that the CF -Chi and CF -PBNp -Chi fabrics
possess high adhesion on the fabrics even when they undergone multiple washes. However, the
percentage of nanoparticles coated on the fabric significantly gets reduced. This implies that
more nanoparticles are evident after 5th wash th ereafter 10th wash. This suggests that more
number of washes leads to alleviation of particl es coating from the fabric surface. This is again
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fostered by the alternations in the calculated thickness of the UC -CF, CF -Chi and CF -PBNp -Chi
fabric s before and after wash (Tab le 2). Nevertheless , concentration of nanoparticles present on
fabric surface is enough to regain their functional properties even up to a maximum usage life of
the fabrics, say 10th wash. In many other applications like wound dressing and biomedical,
antibacterial fabrics are hired for one time use only.
Additionally , the coating thickness of nanoparticles on fabric clearly shows the air
permeability of the fabric s. Air permeability test on UC -CF, CF -Chi including CF-PBNp -Chi
fabrics observed that permeability of the fabric shows 85% of decrease in air permeability as
compared to t hose of UC -CF fabric and CF -Chi fabric that reflected 53% reduction. It is
concluded that the low air permeability in CF-PBNp fabric s is owing to the absorption of
composite materials in between the fibers of the fabric . The air permeability allied to CF-PBNp –
Chi fabric after 5th and 10th washes is improved to 65% and 72%, respectively . Also, the chitosan
coated fabric reflected an upsurge of 60% and 71%, respectively. The increase in air
permeability after 5th and 10th washes is due to alleviation of the coatings from the fabric surface .
The mechanical properties of the CF, CF -Chi and CF -PBNp -Chi fabrics are found and
linked with the functional properties of the fabrics . In this study, the longitudinal and transverse
weaves are analysed with the help of tensile and tear strength for chitosan coated , including the
P. betle chitosan coated fabrics. Prior to washing, the breaking weight of interstices (warp and
weft yarn) values for tensile and tear strengths of CF -PBNp -Chi fabric are increased against CF
and CF -Chi fabrics. Typical ly, the comprehensive tensile and tear strength for the fabrics
specimen CF -Chi/CF -PBNp -Chi are found more than that of CF fabric (Table 2).
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This is owing to the sharing of the load by the coated nanoparticles on the fabrics leading
to an increase in tensile strength. In the similar way, the CF -PBNp -Chi fabric again compliments
a greater bursting strength of UC -CF and CF -Chi fabrics.
The nanoparticles are strongly followed by the surface of cotton fabrics , leading to an
enhan cement in Crease recovery angle (CRA) of fabrics. The CRA of CF -PBNp -Chi fabric
depicts 104 ± 0.76° which is more when compared to CF or CF -Chi fabric.
The observed greater magnitude of crease recovery is a result of higher absorption of
PBNp -Chi nanoparticles on the surface of fabrics without giving any stiffness to the fabric. Apart
from this , the CRA of the CF -PBNp -Chi fabrics is decreased after 5th and 10th washes owing to
the reduction of coating with the increase in number of washings.
The blocking for both UV -A (320-400) and UV -B (290 to 320 nm) is not elucidated for
the CF fabrics [40]. As observed in case of CF-PBNp -Chi fabric, there is a reduction in
transmittance of some specific wavelength . This transmittance is the result of blocking UV -B
and UV -A radiations. When it comes to coated fabrics, it can be observed that the percentage of
UV-B blocking is relatively more when compared to UV -A. The CF -PBNp -Chi fabric shows
great degree of blocking of UV -B radiation when compared to CF-Chi fabrics [41]. Moving
ahead, the blocking rate of UV -radiation for CF -PBNp -Chi fabric s after 5th and 10th washes is
quite decreased when compared to that of the unwashed cotton fabrics. The reduction in blocking
rate of UV -radiation for the coated fa brics elucidated that the subsequent washes entailed to the
alleviation of small amount of coating on the surface of fabrics , which is also evidenced from the
earlier observations [42 -44].

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On the basis of ASTM D6603 standard data, the estimated Ultra V iolet Protection factor (UPF)
value for the fabrics that are utilized in UV -blocking are much greater than 50. The values for CF
and CF -Chi fabrics are lower than 50 when compared to CF -PBNP -Chi fabrics. Table 3,
illustrates t he UPF value for the CF, CF -Chi and CF -PBNp -Chi fabric samples . It is quite
apparent from the Table 3 that CF -PBNp -Chi fabric indicates greater UPF value of about 50,
depicting increased blocking rate of UV -radiation.
The increase in the UV -blocking properties of CF -PBNp -Chi fabric is typically owing t o
the amalgamation of PBNp nanoparticles to colloidal chitosan sol on the surface of cotton
fabrics. Thus , the CF -PBNp -Chi fabrics can impeccably combat the UV -radiation after complete
washing process.
3.3. Antibacterial ass essment of coated fabrics
The analysis of the antibacterial activity of the obtained PBNp nanoparticles, including
CF, CF -Chi and CF -PBNp -Chi fabrics are assessed by cautiously evaluating the diameter of
inhibition zone. The disc encumbered with PBNp nanoparticles reflected the maximum zone of
inhibition as compared to E. coli and S. aureus bacteria at a concentration of 100 mg ml-1.
The antibacterial activities of CF -Chi and CF -PBNp -Chi coated fabrics are investigated
higher area for the zone of inhibition is noticed for CF -PBNp -Chi fabrics that convenes more
inhibitory action ( 25.45 ± 0.34 mm and 31.58 ± 0.06 mm) as compared to CF -Chi fabrics .
However, when it comes to S. aureus , the difference in the magnitude of zone of inhibition is
somewhat high (26.81 ± 0.07 mm and 34.01 ± 0.08 mm) than E. coli .
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Table 2 Physical properties of un -coated and nanoparticles chitosan coated cotton fabrics
Sample code Tensile strength

Warp (kg) Weft (kg) Tear strength

Warp (kg) Weft (kg) Air
Permeability
(cc s-1cm-2) CRA
(W+F)ș Thickness
(mm) Bursting
strength
kg cm-2
UC-CF
CF-Chi
CF- PBNp -Chi 46.1 ± 0.1
47.9 ± 0.1
48.5 ± 0.4 41.3 ± 0.1
42.8 ± 0.1
44.6 ± 0.3
21.2 ±0.1
23.9 ± 0.2
27.1 ± 0.1
17.9 ± 0.1
19.8 ± 0.1
22.1 ± 0.1 97.42 ± 0.4
85.97 ± 0.2
53.32 ± 0.1 98.6 ± 0.2
103.4 ±0.4
104.8 ± 0.7
0.29 ± 0.1
0.33 ± 0.5
0.34 ± 0.1
6.31 ± 0.5
6.62 ± 0.6
7.45 ± 0.2

After 5th wash
UC-CF
CF-Chi
CF- PBNp -Chi 33.2 ± 0.1
35.1 ± 0.1
40.2 ± 0.4 29.2 ± 0.1
32.6 ± 0.2
34.3 ± 0.2
17.3 ± 0.3
21.4 ± 0.2
23.3 ± 0.1
13.6 ± 0.2
14.1 ± 0.1
16.4 ± 0.2 98.47 ± 0.2
65.93 ± 0.1
60.02 ± 0.5
75 ± 0.2
92 ± 0.2
96 ± 0.2 0.27 ± 0.1
0.32 ± 0.5
0.33 ± 0.1 6.30 ± 0.1
6.51 ± 0.1
6.78 ± 0.2
After 10th wash
UC-CF
CF-Chi
CF- PBNp -Chi 26.3 ± 0.3
29.8 ± 0.1
29.1 ± 0.1
20.4 ± 0.4
23.1 ± 0.1
25.2 ± 0.1
15.4 ± 0.4
16.1 ± 0.1
19.5 ± 0.2
10.2 ± 0.4
11.6 ± 0.1
13.5 ± 0.2 99.97 ± 0.1
72.44 ± 0.1
71.13 ± 0.2
45.6 ± 0.2
61.5 ± 0.1
60.8 ± 0.1
0.25 ± 0.1
0.30 ± 0.1
0.32 ± 0.2
6.1 ± 0.1
6.3 ± 0.2
6.6 ± 0.3

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Additionally, in case of UC – CF fabrics, the formation of inhibition is completely absent . A
thorough analysis of the current outcomes shows that CF -PBNp -Chi fabric displays maximum
zone of inhibition against E. coli and S. aureus. The higher antibacterial action of PBNp sample
is obtained owing to the existence of phytochemic al compounds in Piper betle leaves , such as
carbohydrates, alkaloids, flavonoids , phenols, protein and steroids [45]. The above mentioned
bacterial susceptibility investigations coupled by plant extract and herbal nanoparticles shows the
striking medicinal properti es and therapeutic uses of PBNp, nanoparticles , which can be utilized
in medical textile.
3.4. Percentage reduction test (AATCC 100) and washing durability for coated fabrics
Table 3 shows t he quantitative calculation of the antibacterial action for the UC -CF and
CF-PBNp -Chi fabrics after percentage reduction test evaluation . The percentage reduction test
performed on organisms treated with CF -PBNp -Chi fabrics both before and after washing is 99
and 93 %, respectively . In addition to this, for CF -Chi fabrics , it is found 77 % and 70 %,
respectively . The percentage reduction test does not show any evidence for bactericidal activity
of CF fabric. After commencing 5th and 10th wash, the antibacterial activity is decreased owing to
the elimination of herbal nanoparticles from fabric surface as compared to E. coli and S. aureus
bacteria. The antibacterial activity of the CF -PBNp -Chi fabrics after 10 washings is more than 40
%. The wash durability of CF -PBNp -Chi fabrics after 10th wash is found to be more, which
ensures superior functional properties of herbal nanoparticles coated fabrics.

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Table 3 Bacterial reduction percentage of Piper betle chitosan coated fabrics

4. Conclusion
In the bottom -line, herbal nanoparticles are obtained from the shade dried leaves of Piper
betle with the help of ball-milling technique that takes place in three different milling durations
(5, 10 and 15 h). The best sample is collected on the basis of characterisation results for further
coating on the cotton fabrics. Going ahead, it is observed that the coated fabrics show enha nced
UV-protective action at 284 nm. A good amount of nanoparticles placed on the fabric surface are
remained even when it is washed for 10 times . The discovered physical properties of the CF-
PBNp -Chi coated fabric s are much more superior as compared to chitosan -coated or un-coated
fabric materials. The herbal nanoparticles on the cotton fabrics have also show n better
antibacterial activity when compared to S. aureus and E. coli bacteria. From the aforementioned
outcomes , it is clear that the natural phytochemical constituents of P. betle nanoparticles are
good at retain ing their medicinal propert ies and can be coated on cotton fabrics . This, in turn,
enhances various properties , like antibacterial activity, washing durability, and UV -blocking No. of
washes Bacterial reduction percentage (%)
CF-Chi CF- PBNp -Chi UPF value (%)
E. coli S. aureus E. coli S. aureus UC-CF CF-Chi CF-PBNp –
Chi
0 wash 82 ± 0.81 78 ± 0.21 95 ± 0.72 99 ± 0.32 13.9 42.8 55.2
5th wash 70 ± 0.31 60 ± 0.11 81 ± 0.34 83 ± 0.39 11.2 40.1 53.8
10th wash 40 ± 0.14 36 ± 0.11 45 ± 0.26 46 ± 0.12 10.8 39.5 50.8
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ability. Therefore , this study highlights the use of P. betle nanoparticle s coated with CF-PBNp –
Chi fabrics for enhanc ing applications in biomedical and textile industries.
Acknowledgment
The authors acknowledge the financial support provided by Board of Research and
Nuclear Science (BRNS), Mumbai (Sanction no:2013/34/30/BRNS/1127dt.19.9.2013). One of
the authors (Dr. R.S) is thankful to the University Grants Commission (UGC), New Delhi for the
award of Post -Doctoral Fellowship for Women (F.15 -1/2015-17/PDFWM – 2015 -17-TAM -36274
dt.12/10/2015).
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