VISVESVARAYA TECHNOLOGICAL UNIVERSITY

BELAGAVI-590018, KARNATAKA.
(KSCST Sponsored Project)

A PROJECT REPORT ON

“ANTIMICROBIAL ACTIVITY AND FRAGRANCE OF CLOVE BUD ON

WOVEN SILK FABRIC”
By

AMRUTA V PURANIKMATH 4BD16TX047

SAKSHI JAIN 4BD17TX024
SINDHU SHREE M 4BD17TX030
BINDU M N 4BD17TX006

Submitted in the partial fulfillment of the requirement for the award of degree
BACHELOR OF TECHNOLOGY
in
TEXTILE TECHNOLOGY

GUIDE
Dr. S.M. Chandrasekhara
Asst. Professor

2020-2021
DEPARTMENT OF TEXTILE TECHNOLOGY,
BAPUJI INSTITUTE OF ENGINEERING AND TECHNOLOGY, DAVANGERE
 DEPARTMENT OF TEXTILE TECHNOLOGY

2020-2021
PROJECT (17TXP78)

CERTIFICATE

This is to certify that the project work entitled “ANTIMICROBIAL ACTIVITY AND
FRAGRANCE OF CLOVE BUD ON WOVEN SILK FABRIC” carried out by Ms. AMRUTA
V PURANIKMATH, Ms. SAKSHI JAIN, Ms. SINDHU SHREE M, Ms. BINDU M N, a
bonafide student of this institution in partial fulfillment for the award of Bachelor of Technology
in Textile Technology of the Visvesvaraya Technological University, Belagavi during the year
2020-2021. It is certified that all corrections/suggestions indicated for Internal Assessment have
been incorporated in the report and deposited in the departmental library. The Project report has
been approved, as it satisfies the academic requirements with respect to the Project work prescribed
for the said degree.
Project Guide
Dr. CHANDRASEKHARA S. M
M. Tech., Ph.D.
Asst. Professor

Head of the Department Principal

Name of the Examiners Signature with Date

1.
2.

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 ACKNOWLEDGEMENT

It is a matter of great pleasure and privilege for us to present this report of our project
work. We would like to thank our guide Dr. S. M. Chandrasekhara for helping us
throughout the project work. His guidance has been the standing pillar in architecture
of this report. We heartily express our sense of gratitude to Dr. K. B. Ravindra,
Associate Professor & Head of the Department of Textile Technology, for his
support to carry out the project work and for being a helping hand for all the
research-oriented activities related to the work. We would also like to thank Dr.
Manjunath N. S, Head of the Department, Biotechnology, T Chandrakishore
Assistant Professor of Biotechnology, for his support in carrying out the experiment
at the Microbiology laboratory with the cooperation of both faculty and working
staff. We express our respect and gratitude to our principal, Dr. H. B. Aravind, for
his continuous monitoring and fulfilling of all the requirements to carry out the
project work. We sincerely express our gratitude to all the faculties and non-working
staff members of Textile Technology department for being so generous in helping
us carry out the project work, without whom this project work would not have been
possible.

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 BAPUJI INSTITUTE OF ENGINEERING AND TECHNOLOGY
DAVANGERE
DEPARTMENT OF TEXTILE TECHNOLOGY
VISION OF INSTITUTE

To be center of excellence recognized nationally and intentionally, in distinctive areas of

engineering education and research, based on a culture of innovation and invention.

MISSION OF INSTITUTE

BIET contributes to the growth and development of its students by imparting a broad based
engineering education and empowering them to be successful in their chosen field by inculcating
in them positive approach, leadership qualities and ethical values.

VISION OF DEPARTMENT

To be center of excellence in textile, garment and fashion education at the under graduate, post
graduate and doctoral levels, besides serving the society by understanding various innovative
research, industry and social related activities.

MISSION OF DEPARTMET

To impart the basic science, engineering, textile, garment and fashion education for the growth
and development of the students by providing outstanding teaching and learning processes.

To impart practical/research training/internship/projects knowledge through industrial experience.

To empower the students with positive approach and presentation skills by conducting trainings,
workshops and conference.

To inculcate ethical values of various virtues which enable them to become

successful textiles technologists and good citizens of this country.

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 PROGRAMME EDUCATIONAL OBJECTIVES (PEOS)

PEO1: Have a fundamental understanding if basic science, engineering and experimentation skills
required for multidisciplinary domains.

PEO2: Develop expertise in various systems of manufacturing of textiles and fashion. materials
and apply them for new process and product development.

PEO3: Inculcate research capabilities in various textile and fashion technology fields to solve
problems and adopt them in the world of constantly evolving technology.

PEO4: Exhibit entrepreneurship, leadership quality, team work and incorporate societal needs to
practice their profession with high level of legal and ethical responsibilities.

PROGRAM SPECIFIC OUTCOMES (PSOS)

Graduates will have knowledge of science and engineering of textiles leading them leading them
towards successful career. The students will be capable of designing and developing various textile
product controlling and improving the various manufacturing processes and controlling of quality
textile products.

The students will be able to communicate their ideas, findings and knowledge through project
work, technical publications, technical conferences, etc.

Graduates will orient themselves to become successful technocrats who can contribute
technological, industry and societal development and also become successful entrepreneurs.

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 TABLE OF CONTENTS

Serial Contents Page No

No.
I Chapter I 9-11
1.1 Introduction 9-10
1.2 Objectives 11
II Chapter II 12-38
2.1 Literature survey 12-21
2.2 Silk as a textile fiber 22
2.3 Components of Silk 23-25
2.4 Properties of Silk 26-29
2.5 Antibacterial property of clove 29-31
2.6 Application of Antimicrobial finish 31-34
2.7 Antimicrobial finish of silk 34-37
2.8 UV Protection of silk 37-38
III Chapter III 39-51
3.1 Materials and Methodology 39-41
3.2 Geometrical properties of silk fabric 41-42
3.3 Extraction of essential oil from clove 43
3.4 Treatment of silk fabric with alum 44-45
3.5 Application of clove extract 45
3.6 Medium of preparation for the bacteria growth 46-49
3.7 Antibacterial activity of clove extract on silk fabric 49-50
3.8 Washing test 50
3.9 Color strength test 50-51
3.10 Fragrance intensity test 51
IV Chapter IV 52-56
4.1 Results & Discussion 52
4.2 Geometrical properties of silk fabric 53

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4.3 The antimicrobial activity of finished silk fabric and zone of 54-56
inhibition
V Chapter V 57-58
5.1 Conclusion 57-58
6 References 59

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 ABSTRACT

The objective of this study is to explore, evaluate and compare the antibacterial
activity of a Silk woven fabric treated with clove oils extracted from clove buds. The
Clove bud is rich in eugenol, and flavonoid, essential oil that can be used for
antimicrobial activity. The finishing agent, clove oil was extracted by steam
distillation techniques. The antimicrobial activities were evaluated against
Escherichia coli (gram-negative) bacteria by zone of inhibition measurement. Silk
woven fabric finished with clove extract showed strong antimicrobial activity
against Escherichia coli (18-20mm) bacteria. In addition to this, the washing test and
fragrance intensity test, color strength of the woven silk fabric with clove finish was
measured before and after washing and has got the some promising effect. From this
study, it was revealed that clove extract finish has better, durable antibacterial
potential with good fragrance intensity after some washes.

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 CHAPTER I

1.1 INTRODUCTION

The growth of micro-organisms on textile materials inflicts a range of undesirable

effects not only on the textile itself but also on the wearer. These effects often lead
to objectionable odor, dermal infection, allergic diseases, stains and discoloration in
the fabric and product deterioration. The textiles not only act as substrates for
microbial growth but also act as active agents in propagation of microbes. Increasing
public awareness about the risks of microbial infection is a growing demand for
products, which have antimicrobial properties against microbial contamination.
Most of the textiles currently used in hospitals and hotels are conductive to cross-
infection or transmission of diseases caused by micro-organisms, particularly
bacteria and fungi. In view of this, prevention of microbial growth has become
increasingly important and this necessitates the development of clothing that could
provide a desired antimicrobial effect. The finishing of textile materials with
antimicrobial agents is necessary to avoid cross infection by pathogenic micro-
organisms, to control the infestation by microbes, to arrest metabolism in microbes
to reduce odour formation and to protect the textile products from staining,
discoloration and quality deterioration.

A number of antimicrobial agents have been developed to impart antimicrobial

activity to textile goods which are presently being used in the industry. Many of the
commercial antimicrobial agents currently available in the market are synthetic and
may not be environment friendly. Antimicrobial agents on finished textiles may
interfere with the skin’s immune system and its specific defense mechanism. Their
compliance to the regulations imposed by international bodies. The active
ingredients used in antimicrobial finishing need to be effective, have selective
activity towards undesirable microbes, non-toxic, safe, biodegradable and durable to
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wash. Natural bioactive agents with antimicrobial properties have become
increasingly important for bio functionalization of textile fibres because they enable
the production of safe, non-toxic, skin and environmental friendly bioactive textile
products. These antimicrobial compounds, which are mostly extracted from plants
(Clove oil, Aloe Vera, Tea-tree and Eucalyptus oil (EO), Neem, Grapefruit seed,
Tulsi leaf extracts, etc.), include phenolic and polyphenol (Simple phenols, phenolic
acids, quinines, flavonoids, flavones, flavonols, tannins and coumarins), terpenoids,
essential oils, alkaloids, lactins, polypeptides and polyacetylenes. These components
show, not only antimicrobial, but also antioxidant properties.

India has rich biodiversity and there are 300–450 medicinal and dye plants with
antimicrobial composition to be effective candidates in bringing out antimicrobial
textiles. Most of the naturally available herbal extracts are environmental friendly.
Therefore, it is indispensable to need more investigation in the development of eco-
friendly, bioactive antibacterial agents extracted from plants and animals for textile
applications.

In this project the effort is made to finish the woven silk fabric using clove extract
and evaluation of antimicrobial activity is carried out by agar well diffusion method,
and the fragrance test is conducted by subjective method.

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1.2 OBJECTIVES

 Finishing of silk fabrics with clove extract.

 To evaluate the microbial activity on finished fabric sample.

 Estimating fragrance intensity on the finished sample.

 To evaluate the durability of antimicrobial finish after wash.

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 CHAPTER II
2.1 LITERATURE SURVEY

Subrata Das et.al carried out study on Antimicrobial activity and fragrance of
clove bud on woven silk fabrics,

In this paper, we study and evaluate the antibacterial potency of phytochemicals from
syzygium aromaticum (clove) and fragrance intensity on silk woven fabric. The extract
of syzygium aromaticum was characterized using thin-layer chromatography, gas
chromatography--mass spectrometry and fourier transform infrared. Antibacterial assay
was performed with 50% concentration of clove extract against escherichia coli
bacteria. Fragrance of clove treated silk fabric was determined by subjective method.
Relative colour strength was evaluated. Antimicrobial activity, relative colour strength
and intensity of fragrance of clove treated silk fabric are gradually decreased with the
increase in washing cycle.

K. Murugesh Babu et.al carried out study on Bioactive antimicrobial agents for
finishing of textiles for health care products,

Various antimicrobial textile materials are developed using a variety of active agents
which include synthetic antimicrobial agents such as triclosan, metal and their salts,
phenols, quaternary ammonium compounds, and organometallics. Although
synthetic antimicrobial agents effectively inhibit the growth of microbes, most of
them are toxic, can cause adverse effects on human health, and have environmental
issues. Present studies prove that several plant extracts could be effective against
both gram-positive and gram-negative bacteria depending on the type of components
present in the plant extract. Hence, the research on eco-friendly antimicrobial agents
and their application on various textile products gain worldwide importance. Natural

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antimicrobial compounds derived from plants such as neem, tea tree, azuki beans,
aloe vera, tulsi leaves (Ocimum sanctum), clove oil, pomegranate rind, turmeric,
eucalyptus oil, onion skin, and pulp extracts, are being used in the finishing of
textiles. This paper highlights the possibilities of using these bioactive substances
for imparting antimicrobial property to the textiles for developing health care
products.

G Thelagavati et.al carried out study on Application of Prickly Chaff leaves as

herbal antimicrobial finish for cotton fabric used in healthcare textiles,

Prickly Chaff leaves are reported to have antimicrobial properties. An innovative

approach was made utilize this eco-friendly and renewable source for production of
microbial resistant fabric. The chemical nature of the leaf extract was determined
using HPLC and extraction of active substance from the leaves was done using
methanol. The method of application of this herbal extract on cotton fabric using
citric acid as cross-linking agent and the process variables have been standardized
using box and Behnken three level three variable experimental design. The
antimicrobial activity of the finished fabric based on optimized process parameters
was assessed against bacteria that normally exist in the textile environment, the
finished cotton fabrics showed the bacterial reduction percentage.

Erdogan Ceylon et.al carried out study on Antimicrobial Activity of Spices,

Many of the spices and herbs used today have been valued for their antimicrobial
effects and medicinal powers in addition to their flavor and fragrance qualities. Most
of the forbore bacterial pathogens examined were sensitive to extracts from plants
such as cinnamon, clove, garlic, mustard, onion and oregano. me antimicrobial
compounds in spices and herbs are mostly in the essential oil fraction. The Gram-
positive bacteria were more sensitive to the antimicrobial compounds in spices than

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Gram-negative bacteria. The extent of sensitivity varied with the strain and
environmental conditions imposed. Certain spices can have a direct effect on the rate
of fermentation by stimulating acid production in starter cultures. Phenols, alcohols,
aldehydes, ketones, ethers and hydrocarbons have been recognized as major
antimicrobial components in spices. me antimicrobial activity and modes of actions
of spices and their major antimicrobial components are reviewed.

L. A. Shelef et.al carried out study on Antimicrobial Effects of Spices,

There is a renewed interest in the antimicrobial properties of spices. In vitro activities

of several ground spices, their water and alcohol extracts, and their essential oils
have been demonstrated in culture media. Studies in the last decade confirm growth
inhibition of gram positive and gram negative food borne bacteria, yeast and mold
by garlic, onion, cinnamon, cloves, thyme, sage and other spices. Effects in foods
are limited to observations in pickles, bread, rice, and meat products. In general,
higher spice levels are required to effect inhibition in foods than in culture media.
Fat, protein, and water contents in foods affect microbial resistance as does salt
content. Very few studies report on the effect of spices on spores, and on microbial
inhibition in conjunction with preservatives and food processes. Of the recognized
antimicrobial components in spices, the majority are phenol compounds with a
molecular weight of 150 to 160 containing a hydroxyl group. Eugenol, carvacrol and
thymol have been identified as the major antimicrobial compounds in cloves,
cinnamon, sage and oregano.

Haiying Cui et.al carried out study on Antimicrobial mechanism of clove oil on
Listeria monocytogenes.

Clove oil is widely used in food preservation, very little is known about its possible
antibacterial mechanism. In order to reveal its antibacterial mechanism, the

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inhibitory effect of clove oil on Listeria monocytogenes and its effect on cell
membrane, respiratory metabolism, and its interaction with DNA were investigated,
respectively. The results showed the treatment of clove oil can cause the leakage of
three biological macromolecules (protein, ATP and DNA) and the reduction of two
intracellular enzymes (β-galactosidase and AKP) activities, indicating that clove oil
has an effect on cell membrane permeability. The inhibition rate and superposition
rate test confirmed that the pathway of clove oil affecting the respiratory metabolism
of Listeria monocytogenes is TCA pathway. The treatment of clove oil can reduce
the activity of three key enzymes (isocitrate dehydrogenase, citrate synthase and α-
ketoglutarate dehydrogenase) in TCA pathway, and affect the content of metabolites
in the pathway. The results of ultraviolet absorption spectroscopy showed that
eugenol, the main component of clove oil, can change the structure of

DNA via the formation of eugenol-DNA chimera.

G. A. Ayoola et.al carried out study on Chemical analysis and antimicrobial

activity of the essential oil of Syzigium aromaticum (clove).
In this paper we study about steam distillation of the dry flower buds of Syzigium
aromaticum (clove) yielded 7% (w/w) of the pure light yellow oil. Gas
chromatography-mass spectrometry (GC-MS) analysis of the oil revealed that the
components were eugenol, caryophyllene, eugenol acetate and alpha-humelene, with
eugenol being the main component. The antimicrobial sensitivity of the volatile oil
against some Gram-negative bacteria (Escherichia coli ATCC 35218, Escherichia
coli, Klebsiella pneumoniae, Salmonella paratyphi, Citrobacter spp. and
Enterobacter cloacae), a Gram-positive bacterium (Staphylococcus aureus
ATCC25923), and a fungus (Candida albicans) showed a broad spectrum of
activity. The minimum inhibitory concentration (MIC) was determined for each
organism as 2.4, 1.6, 0.27, 0.016, 0.23, 1.63, 0.73 and 0.067 mg/ml for S. aureus

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ATTC 25923, E. cloacae, S. paratyphi, K. pneumoniae, E. coli ATTC 35218, E. coli,
Citrobacter spp. and C. albicans, respectively. Antioxidant screening of clove oil
with 2,2-diphenylpicryl-hydrazyl radical (DPPH) was positive, indicating the
presence of free radical scavenging molecules which can be attributed to the
presence of eugenol, a phenolic compound.

Gao Hongfang, et.al carried out study on Controllable preparation and

mechanism of nano-silver mediated by the micro emulsion system of the clove
oil.
This paper carried out study on A clove oil micro emulsion system for preparing
silver nanoparticles (Ag-NPs). The effects of the clove oil concentration and the
reaction time on the formation of Ag-NPs were investigated. The synthesized Ag-
NPs was characterized by UV–Vis, X-ray diffraction and high resolution
transmission electron microscopy. It was found that the clove oil acted as a reducing
agent and a stabilizer in the formation of silver nanoparticles. The mechanism was
that Ag+ was adsorbed on the oil–water interface by the active groups of the clove
oil in the microemulsion in which it was then reduced to nano-Ag. Furthermore,
microspheres were formed on the surface of nano-Ag through complexation of
macromolecular substances. Alkyl and aromatic hydrophobic groups that were
distributed on the surface of nano-Ag were driven into the clove oil phase under the
promotion of Gibbs free energy. Thus, Ag nano-particles with good dispersibility
and stability were obtained. The infrared spectrum shows that the outer layer of the
nanoparticles coated with active groups of clove oil, which can give the silver
nanoparticles better antibacterial. Ag nano-particles of spherical, polygonal, and rod-
like shapes were achieved by controlling the initial clove oil concentration and the
reaction time.

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L. B. Bullerman, et.al carried out study on Inhibition of growth and aflatoxin
production by cinnamon and clove oils, cinnamic aldehyde and eugenol.
This study determines the specific components of cinnamon and cloves that may be
effective against mold growth and toxin production. The effects of cinnamon oil,
clove oil, cinnamic aldehyde and eugenol on growth and aflatoxin production by
Aspergillus parasiticus were studied using yeast-extract sucrose broth as the
substrate. All four substances inhibited mold growth and subsequent toxin
production. Cinnamon and clove oils were inhibitory at 200-250 ppm, cinnamic
aldehyde at 150 ppm and eugenol at 125 ppm. Since cinnamic aldehyde and eugenol
are the respective major components of cinnamon and clove oils, it was concluded
that these are the major active antifungal ingredients of these two essential oils. The
inhibitory effect of these substances was judged to be inhibition of growth rather
than of toxin production. When growth occurred after a delay, aflatoxin production
occurred when the cultures reached secondary metabolism. Given sufficient time,
cultures which were inhibited initially, but which subsequently grew, produced toxin
levels equivalent to control cultures. Levels of the oils above 250 ppm and of
cinnamic aldehyde and eugenol above 200 ppm completely inhibited mold growth,
or permitted only a small amount of growth that never reached secondary
metabolism and never produced aflatoxins during the time of this study.

K. Murugesh Babu, et.al carried out study on Silk Fibroin Coated Antimicrobial
Textile Medical Products.
In this study an attempt has been made to develop eco-friendly natural fibre based
medical products such as baby’s diaper, sanitary napkins and surgical wound
dressings. These products were developed by coating natural silk fibroin protein
extracted from natural silk fibre. An effective technique has been used for extraction
of silk fibroin protein from raw silk by successive methods of degumming,
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dissolution and dialysis. In addition, the silk fibroin protein was combined with
certain natural oils such as neem, clove, eucalyptus which were found to possess
anti-microbial property. This protein/oil mixture in the ratio of 10:1 was applied to
impart anti-microbial property to the products. Medical products incorporating
natural fibres like organic cotton, sterilized cotton and bamboo coated with the
natural silk fibroin protein and natural anti-microbial oils demonstrate excellent
holding capacity, wicking behavior compared to commercially available products.
All the products exhibit very good anti-microbial property which is needed for the
medical product.

Gang Li, et.al carried out study on Silk-Based Biomaterials in Biomedical

Textiles and Fiber-Based Implants.
In this paper a biomedical textiles and fiber-based implants (BTFIs) have been in
routine clinical use to facilitate healing for nearly five decades. Amongst the variety
of biomaterials used, silk-based biomaterials (SBBs) have been widely used
clinically viz. sutures for centuries and are being increasingly recognized as a
prospective material for biomedical textiles. The ease of processing, controllable
degradability, remarkable mechanical properties and biocompatibility have
prompted the use of SBBs for various BTFIs for extracorporeal implants, soft tissue
repair, healthcare/hygiene products and related needs. The present Review focuses
on BTFIs from the perspective of types and physical and biological properties, and
this discussion is followed with an examination of the advantages and limitations of
BTFIs from SBBs. The Review covers progress in surface coatings, physical and
chemical modifications of SBBs for BTFIs and identifies future needs and
opportunities for the further development for BTFIs using SBBs.

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Ashutosh Bandyopadhyay, et.al carried out study on Silk: A Promising
Biomaterial Opening New Vistas Towards Affordable Healthcare Solutions.
A study on substantial progress in biomaterial research over the years has culminated
in revolutionary technological advancements in the healthcare domain. This has
triggered the quest for affordable healthcare solutions with focus on sustainable
biomaterials with versatile applications endowed with green fabrication strategies.
Silk as a biopolymer has garnered special attention which can largely be attributed
to the excellent material properties of silk in addition to its affordability and resource
ability. Silk fibroin from various silkworm and spider species and sericin from
various silkworm species have been researched for their potential applications in the
healthcare industry such as tissue engineered grafts, cancer therapeutics, high-
throughput tissue-on-chip models, food preservatives, biomedical imaging, bio
sensing, biomedical textiles, implants, cosmetics and bioremediation products. The
present review mainly focusses on the various sources of silk fibroin and its relevant
properties that have been conferred to it by nature. Moreover, recent developments,
progress and prevalent modalities of healthcare industry that involve the application
of silk fibroin and sericin have been outlined in the present review.

Gregory H, et.al carried out study on Silk-based biomaterials.

Silk from the silkworm, Bombyx mori, has been used as biomedical suture material
for centuries. The unique mechanical properties of these fibers provided important
clinical repair options for many applications. During the past 20 years, some
biocompatibility problems have been reported for silkworm silk; however,
contamination from residual sericin (glue-like proteins) was the likely cause. More
recent studies with well-defined silkworm silk fibers and films suggest that the core
silk fibroin fibers exhibit comparable biocompatibility in vitro and in vivo with other
commonly used biomaterials such as polylactic acid and collagen. Furthermore, the
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unique mechanical properties of the silk fibers, the diversity of side chain
chemistries for ‘decoration’ with growth and adhesion factors, and the ability to
genetically tailor the protein provide additional rationale for the exploration of this
family of fibrous proteins for biomaterial applications. For example, in designing
scaffolds for tissue engineering these properties are particularly relevant and recent
results with bone and ligament formation in vitro support the potential role for this
biomaterial in future applications. To date, studies with silks to address biomaterial
and matrix scaffold needs have focused on silkworm silk. With the diversity of silk-
like fibrous proteins from spiders and insects, a range of native or bioengineered
variants can be expected for application to a diverse set of clinical needs.

Alhayat Getu, et.al carried out study on Technical Fabric as Health Care
Material.
This study is to expose the achievements of advance application of textile material.
Previously textiles are only used as normally wound care products, diapers, braces,
prostheses and outhouses, wipes, breathing masks, bedding and covers, ropes and
belts etc. but the technology has been upgraded. Textile materials and products that
have been engineered to meet particular needs are suitable for many applications as
well as medical and surgical application in which a combination of strength,
flexibility, and sometimes moisture- and air-permeability is required. Materials used
include monofilament and multifilament yarns, woven, knitted, and nonwoven
fabrics, and composite structures. The applications are many and diverse, ranging
from a single-thread suture to the complex composite structures used for bone
replacement, and from the simple cleaning wipe to the advanced barrier fabrics used
in operating rooms. Although textile materials have been widely adopted in medical
and surgical applications for many years, new uses are still being found. Research
utilizing new and existing fibres and fabric-forming techniques has led to the
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advancement of medical and surgical textiles. At the forefront of these developments
is absorbency, tenacity, flexibility, softness, or biodegradability.

S. Nivedita, et.al carried out study on Biomedical Applications of Mulberry Silk

and its Proteins: A Review.
The study of silk fibre for aesthetic purposes and also It has been used for making
surgical sutures led to rediscovery of silk’s biological properties has led to new areas
of research and utilization in cosmetic, health and medical fields. The silk proteins,
fibroin and sericin are processed into biomaterials because of bio-compatibility, bio-
degradability, excellent mechanical properties, thermos tolerance and UV protective
properties. Silk proteins could be obtained as pure liquids and regenerated in
different forms suitable for tissue engineering applications. This paper presents some
of the biomedical products and biomaterials made from native, degraded and
regenerated silk and their fabrication techniques.

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2.2 SILK AS A TEXTILE FIBER

Silk is a natural fibre used mainly for aesthetic purposes. It has also been used for
making surgical sutures for centuries. The recent rediscovery of silk’s biological
properties has led to new areas of research and utilization in cosmetic, health, and
medical fields. The silk proteins, fibroin, and sericin are processed into biomaterials
because of bio-compatibility, bio-degradability, excellent mechanical properties,
thermotolerance, and UV protective properties. Silk proteins could be obtained as
pure liquids and regenerated in different forms suitable for tissue engineering
applications.

Native silk is a thin filament with a very smooth surface creating a good silk fabric
thus coating of antimicrobial finish on this surface gives us the best result in the
medical textile field. These include knitted socks, gloves, neck braces for
spondylitis, knee and wrist bands, and innerwear for treatment and prevention of
allergies, skin diseases, and infections (eczema and mycosis) and protect skin against
loss of water, which can aggravate skin dryness. Woven and non-woven silks are
used to make antimicrobial respiratory masks, gauze pads, and bandages. It is
enhanced by an antibacterial finish on the surface of the fabric. The smooth silk fibre
avoids mechanical friction and irritation and can be applied in direct contact with
damaged or wounded skin. Fibroin-based wound dressing that accelerates healing,
which allows peeling-off without damaging the newly formed skin has been
developed.

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2.3 COMPONENTS OF SILK

Silk, a structural protein represents a distinct class of biocompatible and green

polymers. It has been focused upon in biomedical research pertaining to its
biodegradability, low immunogenic response, and easier processability. Silk can be
credited as one of the most ancient materials known to human beings which have
been documented for its use as a medical suture, as early as 131–211 A.D by Greek
physician Aelius Galenus. The US Pharmacopeia (USP) classifies the conventional
silk sutures which are still in practice today as non-degradable and non-adsorbent,
primarily because of the wax coating which protects the silk fibroin (SF) from
proteolytic digestion. In addition to silk being a prime candidate for medical
applications such as tissue engineering and drug delivery applications, silk has also
been gaining prominence in new frontiers. The mild processing of silk fibres to
obtain aqueous-derived regenerated silk fibroin aids in the feasibility of fabricating
silk fibroin-based photonic devices or biosensors for various biomedical
applications. Silk has also been proven to be an effective stabilizing agent extending
the shelf life of fruits and bio-pharmaceutical agents like vaccines and antibiotics, it
must be physiologically acceptable by the body and broken down to get assimilated
or safely eliminated from the body without eliciting any adverse reaction. Silk meets
these criteria owing to its biocompatibility and biodegradability traits. Moreover, the
greener approaches utilized to process the silk protein from the cocoon and silk
glands further endow it as an ideal candidate for various biomedical applications.
The present review of silk fibroin-based healthcare materials focuses on the diversity
and sources of silk fibroin as well as on the components of silk fibroin that elicit the
necessary properties for hitherto utilization of silk as an ideal biomedical material.
Following the exploration of silk as a biomedical material, the represents emphasizes
the real-world, potential, futuristic as well as prototype technologies that comprise

23
silk as a major component. These healthcare applications of silk have been broadly
spanned into tissue engineering, cancer therapeutics, tissue-on-chip models, food
technology, biomedical sensors, imaging and electronics, cosmetics, biomedical
textiles, and bioremediation.

2.3.1 SILK FIBROIN

The Lepidoptera larvae of Bombycid family spins twin delicate filaments of silk
fibroin enwreathed by silk sericine exhibiting a triangular smooth cross-section as
observed at the microscopic level. The mulberry silk fibroin consists of an H-chain,
an L-chain and a glycoprotein (fibrohexamerin fhx/P25), assembled in the molar
ratio of 6:6:1. The H-chain and L-chain are linked by disulphide linkage and six such
hetero-dimers get associated with a single P25 at the H-chain moiety through
hydrophobic interactions. The N-terminal has glycosylation containing mannose and
glucosamine residues. Poly-(glycine-alanine) repeats forms the primary repeat
sequences which accounts for the ß-sheet crystallite regions observed in the silk. The
crystallite regions are embedded between amorphous α-helical regions, and these
crystallite regions exhibit strong interchain interactions through hydrogen bonding
contributing to the silk’s superior strength. The silk spun by Lepidoptera larvae
belonging to the Saturniidae family is very much distinct from the mulberry silk.
The fibroin L-chain (flc) and fhx/P25 are absent in the non-mulberry silk varieties.
Evolutionarily the gene encoding for fhx/P25 is thought to represent a paralog of
gene(s) which might have adopted new functions other than the fibre formation as
noticed in Bombycid family. Hence, the genes encoding for flc and fhx/P25 have
been lost in the ancestors of Saturniidae family. The H-chain forms homodimers
which constitutes the fibroin core in the silk fibre. Unlike the poly-(glycine-alanine)
repeats which constitute the crystalline domains of mulberry silk, the non-mulberry
silk possesses poly-(alanine) repeats. The silk fibroin arising from the Saturniidae

24
larvae spinneret also consist of two fibroin monofilaments enveloped by silk
sericine, similar to B. mori18. However, the monofilaments of non-mulberry silk
disorient during fibre formation, due to the inability of H-chains to pack closely
owing to the higher percentage of bulky side chains present in the silk fibroin. Thus,
the non-mulberry cocoons appear more fibrous (flattened cross-section of silk
fibroin) in nature which is evident from the density difference between the mulberry
and non-mulberry silk fibres.

2.3.2 SILK SERICIN

Silk sericin, an amorphous glycoprotein is produced in the middle silk gland and
constituted primarily by serine (32–34%), aspartic acid (14– 16%)25 and other
amino acids such as histidine, threonine, tyrosine and glutamic acid26. Sericin exists
as random coil conformation and the molecular weight of sericin varies between 10
and 350 kDa. The fraction of sericin obtained solely depends upon the extraction
protocol. For instance, hydrolysates of sericin obtained through plain water boiling
are mostly low molecular weight (10–20 kDa), while other methods yields high-
molecular-weight fractions. Five different fractions of sericin from B. mori have
been reported, namely ser-1, ser-2, ser- 3, ser-4 and ser-5 ranging between 24 and
400 kDa. The non-mulberry silk sericin varies from the mulberry silk varieties, for
instance, the glycine content is lower in A. Though sericin is considered as a by-
product and deemed as waste in sericulture industry, it has attracted immense interest
from the cosmetic and pharmaceutical applications owing to its antibacterial,
antioxidant, anticoagulant and wound healing properties. The enhanced
environmental stability of silk fibers in comparison to globular proteins is due to the
extensive hydrogen bonding, the hydrophobic nature of much of the protein, and the
significant crystallinity. Silks are insoluble in most solvents, including water, dilute
acid and alkali.

25
2.4 PROPERTIES OF SILK

Over the centuries, silk has been regarded as a highly valued textile fibre. Its qualities
of strength, elasticity, softness, absorbency, affinity for dyes and adaptability to
various forms of twisting continue to meet various applications. Because of its high
(tensile) strength, lustre, durability and ability to bind chemical dyes, silk is still
considered a leading textile material. Despite facing keen competition from man-
made fibres, silk has maintained its supremacy in the production of luxury apparel
and other high-quality goods. Silk fibres display unusual mechanical properties: they
are strong, extensible and mechanically compressible. Silk is rightly called the
'queen of textiles' for its lustre and feel. Silk's natural beauty and proper ties of
comfort in warm weather and warmth during colder months have also made it ideal
for high-fashion clothing. As a result, there is significant research into the artificial
production of silk fibres.

2.4.1 PHYSICAL PROPERTIES

I) Moisture Absorption

The silk filament consists of 70-75% solid matter and the rest 25-30% is hollow
(blank). Thus, it absorbs plenty of air, salts, acids, alkalis and is easy to dye. Silk is
porous, absorbs water molecules from the atmosphere and also when immersed in
water. The diameter of the fabric swells up to 16-18%, while the weight goes up by
30-35%. The strong moisture and water absorption capacity is an important property
for clothing material. Many physical and mechanical properties undergo
considerable change due to the absorption of humidity and water by the silk material.
The standard rate of water content has been fixed at 11%.

The humidity absorption of the cocoon filament is critical in preventing molding and
also changes in the cocoon. The hygroscopic properties of raw silk are of great

26
importance from a commercial point of view. The regain value of silk is taken as
11% but it must be borne in mind that the regain for defined conditions of RH% and
temperature depends on whether the equilibrium state is approached by up-take
(absorption) or loss (desorption) of water. The water content of raw silk is generally
determined by heating the material in an oven at 120° C.

II) Specific Gravity

Silk has a specific gravity between 1.36 to 1.39 g/cm² with a specific volume of 0.75
cm²/g and is lighter than wool, cotton and even artificial silk.

III) Tenacity and Elongation

The load taken by a strand of silk filament or a strand of raw silk until it breaks due
to elongation under specified conditions is termed as absolute tenacity. There may
be slight variation in tenacity even when the same size is being used.

The elongation of the silk filament is 18-23%. The elongation of the wet cocoon
filament is more (28-29%), but its tenacity is less. The tenacity of cocoon when it is
dry is 3.2-3.4 g/d and is 3 when it is wet.
The tenacity and elongation of silk varies with the different races of silkworm, the
cocoon layers and the rearing season. Silk loses 34-40% of its strength and its
elongation increases by 40-60% when the threads are wet. The tenacity per denier
of a given breed does not appear to be affected by the Locality of rearing. It has been
observed that degummed silk shows a higher tenacity than the undegummed silk as
sericin does not contribute to tenacity in proportion to its weight. A filament of silk
is stronger than an equal filament of steel.

27
IV) Thermal Conductivity
Thermal conductivity of the silk fibre is expressed in terms of the rate of heat
conductance (thermal conductivity co-efficient). The thermal conductivity is. poor
and the specific heat is higher than other natural fibres and thus silk is warm in winter
and cool in summer. This is due to the fact that the cocoon filament is a porous fibre
having numerous vacuum spots capable of accommodating large quantity of air.

V) Electrical Conductivity
Silk is a bad conductor of both heat and electricity. In dry conditions its resistance
is extremely high. Thus, it is used for making electric insulators for insulating
electric wires. With increase in the moisture regaining rate, its resistance against
electricity falls continuously and finally it becomes a good conductor.

2.4.2 CHEMICAL PROPERTIES

I) Action of Alkalies
Silk is more sensitive to alkali than to acids. Dilute alkali will not react on silk fibre
but dissolves only sericin. When the silk fibre is boiled in a weak alkaline soap
solution for a long time, a part of the fibroin will also be dissolved. The strong alkali
solution dissolves sericin damaging the silk fibre. The damage to the fibre will be
higher when pH, temperature and duration are higher.
II) Action of Acids
The resistance of silk with respect to acid is stronger compared to alkali. The amount
of dissolution of sericin is low when it comes in contact with weak acids. Cocoon-
filament will swell quickly and disintegrate when it comes in contact with strong
acids.

28
III) Action of Sun Light
The silk is sensitive to sunlight, for the ultra-violet rays in the sunlight makes the
silk fragile. Silk in the form of yarn is readily damaged when exposed to sunlight.
Sunlight and moisture are the accelerating factors in the tendering of silk by day
light. Silk is more susceptible to exposure to atmospheric conditions than cotton,
linen and jute. Silk has been found to be the fibre most sensitive to light damage.
This light sensitivity of silk was confirmed by exposure to ultra-violet light of wave
length less than 360μ by many researchers.

2.5 ANTIMICROBIAL PROPERTIES OF CLOVE

Although clove oil is widely used in food preservation, very little is known about its
possible antibacterial mechanism. In order to reveal its antibacterial mechanism, the
inhibitory effect of clove oil. The discovery and development of antibiotics have led
to a dramatic improvement in the ability to treat infectious diseases and is among the
major advances of the 20th century. Unfortunately, development of effective
antibacterial agents has been accompanied by the emergence of drug-resistant
organisms due to the irrational and overuse of antibiotics, failure to complete a
course of treatment, genetic versatility of microbes and horizontal transfer of
resistant genes among bacterial species. All the mentioned factors diminish the
clinical effectiveness of antibiotics. therefore, a need for continuous search of a new,
effective and affordable antimicrobial agent. In recent times, there has been renewed
interest on plants as sources of antimicrobial agents due to their use historically and
the fact that a good portion of the world’s population, particularly in developing
countries, rely on plants for the treatment of infectious and non-infectious diseases.
Antiseptic properties of plant volatile oils have been recognized since antiquity.
Indeed, clove oil is most popular and commonly used as an anesthetic in the relieve

29
of toothache in dentistry. It is also used as a carminative, serves as a preservative in
herbal recipes, signifying possible antimicrobial properties. A green environmental
protection, the way of synthesizing clove oil, provides a helpful reference for
preparing medical substances without using toxic reducing and dispersing chemical
reagents. In addition, the strong antibacterial properties of the prepared clove oil
fortify their application in biomedical and other antimicrobial materials.

2.5.1 ACTIVE COMPOUNDS COMPOSITION IN CLOVE

Biological Active Compounds in Clove Which Have Antimicrobial Properties

1. Anethole
2. Benzaldehyde
3. Carvone
4. Caryophyllene
5. Caryophyllene
6. Cinnamaldehyde
7. Ellagic acid
8. Eugenol
9. Eugenyl acetate
10.Furfural
11.Gallic acid
12.Kaempferol
13.Linalool
14.Methyl eugenol

The essential oils of clove are composed of a complex composition of compounds.

Bioactive ingredients in clove such as sulfides, thiols, terpenes and their derivatives,
phenols, glycosides, alcohols, aldehydes, and esters have been investigated for their

30
properties. Although some phytochemicals have been identified, the majority of the
antimicrobial components of spices are phenol compounds with a hydroxyl group (-
OH). Presence of -OH group was found to be responsible for antimicrobial and anti-
inflammatory properties. Phenolic compounds isolated from clove were the most
effective against all bacterial growth. Steam distilled essential oil yielded a mixture
of compounds. The most inhibitory compounds in the oils against bacteria were
phytochemicals such as alkaloids, flavonoids, tannins, terpenoids, saponins.

The phenolic compounds such as carvone, eugenol and Caryophyllene are very
active antimicrobial agents. The essential oil fraction is particularly high in cloves
and eugenol comprises 95% of the fraction that the high tannin content in cloves
(10-13%) provides additional antimicrobial effects. The antimicrobial property of
clove has protection against five pathogens, i.e., E. coli, E. coli 0157:H7, S.
Typhimurium, L. monocytogenes and V. vulnificus. Thus, clove have potent
antibacterial effect against the bacterial pathogens which are harmful.

2.6 APPLICATION OF ANTIMICROBIAL FINISH

The textile materials and clothing are susceptible to microbial attack because they
provide the basic requirements for microbial growth. The materials made of
cellulose and protein natural fibres provide moisture, oxygen, nutrients and
temperature for bacterial growth and multiplication. This often results in
objectionable odour, dermal infection, product deterioration allergic responses and
often similar diseases.

The control of microorganisms (e.g., bacteria, mildews moulds, yeasts and viruses)
on textile fabrics extends to diverse areas such as hospitals, the environment and
everyday households. Neither natural nor synthetic fibres have resistance to

31
microorganisms; thus various antimicrobial finishes and disinfection techniques
have been developed for all types of textiles.

2.6.1 The functions of antimicrobial finishes applied to textiles are

(a) to control the spread of disease and the danger of injury-caused infection

(b) to control the development of odour from perspiration, stains and dirt on textile
materialsand

(c) to control the deterioration of textiles, particularly fabrics made of natural fibres
caused by mildew

Antimicrobial finishes protect the fibre from microbial attacks and protect the owner
against transfer of pathogenic germs. Silk is relatively resistant to attack by mildew,
fungi and other bacteria. but if stored in moist conditions, then damage can occur.
The fungal resistance can be improved by N(2,2'dichlorvinyl) salicylamide and a
0.01%-0.25% solution of benzalkonium chloride. This increases the resistance of
silk to mildew during long storage (up to 2 years) even at high relative humidity.

Silk is a biocompatible biopolymer applicable in biomedical and biotechnological

devices. In this field, the assignment of antimicrobial activity has great importance.
However, a general problem related to all antimicrobial chemicals is to find the
balance between high biocide activity and the requirements of safe handling,
including nontoxic finish processes, and nontoxicity to humans at usual
concentrations and environmental demands. Antimicrobial agents that have been
used industrially in recent years include quaternary ammonium salts, metal salts
solutions and antibiotics. Unfortunately, some of these agents are toxic and
ineffective, which leads to special attention for 'green chemistry' from worldwide
research. Chitin, the second most abundant biopolymer in nature next to cellulose,
is a high molecular weight linear polymer of 2-acetamide-2-deoxy-D-glucopyranose
32
units linked by 1, 4-glucosidic bonds. Chitosan is a deacetylate derivative of chitin,
which is characterized by its nontoxicity, biodegradability and compatibility with
other ingredients. Previously that D-glucosamine hydrochloride (chitosan monomer)
does not show any growth inhibition against several bacteria in which chitosan is
effective. The antimicrobial activity of chitosan is, therefore, related to not only its
cationic nature, but also to its chain length. treated cotton and silk fabrics with
chitosan of low, medium and high molecular weights (210,000, 100,000, 1800,
respectively) at different concentrations. The padded samples were dried at 100°C
for 3 minutes, followed by curing at 150°C for 3 minutes, then washed and dried.
This was followed by screen printing using a natural dye extracted from prickly
pears. Thus, the obtained fabrics were monitored for nitrogen, colour strength after
dyeing/printing and antimicrobial activity. The results signify that the chitosan
concentration and its molecular weight determine the magnitudes of both the
antimicrobial activity and colour strength of the printed fabric. The silk pretreated
with chitosan followed by dyeing with natural dyes assumes high colour values, the
effects of chitosan on dyeability and the antibacterial properties of silk yarns against
two kinds of bacteria: 'Staphylococcus aureus' and 'Escherichia coli' were
investigated. The treated silk samples were found to have an antibacterial potential
due to the antibacterial property of chitosan. SEM photo graphs reveal the deposition
of chitosan on the treated yarns.

Chitosan is used in the textile field. It is applied by wet thermal curing involving
relatively high temperature with energy consumption, costs and possible fabric
degradation; moreover, the addition of toxic reagents, such as glutaraldehyde, is
required as a cross-linking agent. In UV curing, however, radical species are
generated by the interaction of UV light with a suitable photo-initiator, which
induces the curing reaction of reactive monomers and oligomers at low temperature

33
and quickly, with a lower environmental impact and lower process cost than thermal
process.

2.7 ANTIMICROBIAL FINISH OF SILK

Silk samples are prepared and silk hand characteristics are maintained. An
antimicrobial activity test was performed according to ASTM E2149-01 method,
using microbes. A test on treated samples revealed the antimicrobial efficiency, with
a complete elimination of microorganism. Silk materials are luxurious and have
many excellent qualities, such as high luster, comfort in wearing, fine and smooth
texture, high silky softness and excellent drape ability; hence it needs little finish to
improve its aesthetic properties. However, it does lack several functional properties.
For example, silk materials are prone to creases and wrinkles. It gets easily stained
and is susceptible to microbial attack and ultraviolet light. It has a high water
absorbency, hence it has poor oil and water repellency. Silk is also flammable.
Despite of all these deficiencies, one must be very careful performing physical and
chemical treatments on silk, as this delicate fibre may easily lose its luster and/or
softness for which it is so attractive. Therefore, the processing of silk is a very skillful
job. Until recently the processing of silk was restricted among limited small-scale
skillful processors, and classical finishing methods were in vogue. However, in
recent years, diversified uses of silk materials have compelled processors to search
for newer processes and chemicals. On the other hand, the use of chemicals is getting
more and more restricted due to objections from an environmental point of view. In
spite of the above fact, many newer chemicals have proved to be promising for the
finishing of silk.

Silk-based healthcare and hygiene biomedical textiles have been widely used in
clinical applications for the operating theatre and the hospital ward for hygiene, and
the care and safety of patients and staff. These systems include fiber-based woven
34
materials. Silk-based fabrics used in the operating theatre include surgeon’s gowns,
caps and masks, patient drapes, and cover cloths of various sizes. The advantages of
silk-based fabrics include softness, biocompatibility, mechanical performance, and
multi-functions such as antibacterial properties of the silk fabric.

2.7.1 PROPERTIES OF AN EFFECTIVE ANTIMICROBIAL FINISH

The growth rate of microbes can be astoundingly rapid. The bacteria population, for
example, will double every 20 to 30 min under ideal conditions (36-40 °C or 77-98
°F, pH 5-9). At this rate, one single bacteria cell can increase to 1 048 576 cells in
just 7 hours. Therefore, antimicrobial finishes must be quick acting to be effective.

In addition to being fast acting, a number of other important criteria can be listed for
antimicrobial finishes. The antimicrobial must kill or stop the growth of microbes
and must maintain this property through multiple cleaning cycles or outdoor
exposure. The antimicrobial must be safe for the manufacturer to apply and the
consumer to wear. The finish must meet strict government regulations and have a
minimal environmental impact. The antimicrobial finish must be easily applied at
the textile mill, should be compatible with other finishing agents, have little if any
adverse effects on other fabric properties including wear comfort, and should be of
low cost.

2.7.2 MECHANISMS OF ANTIMICROBIAL FINISHES

Despite the long list of requirements, a variety of chemical finishes have been used
to produce textiles with demonstrable antimicrobial properties. These products can
be divided into two types based on the mode of attack on microbes. One type consists
of chemicals that can be considered to operate by a controlled-release mechanism.
The antimicrobial is slowly released from a reservoir either on the fabric surface or
in the interior of the fibre. This 'leaching' type of antimicrobial can be very effective

35
against microbes on the fibre surface or in the surrounding environment. However,
a major factor that has stimulated interest in antimicrobial finishes using natural
sources has been the current vogue that promotes natural and eco-friendly lifestyle.
In addition, textile consumers are now becoming much more aware of the deleterious
effects that microorganisms may have upon textiles and human hygiene. In
particular, the medical textile sector has welcomed the greater applicability of
antimicrobial finishes to stem the possibility of infections arising from the presence
of microorganisms. While there has been increasing awareness of hygiene issues and
greater consumer demands for hygiene products there is also the wider issue of the
presence of antimicrobial that may come into contact with human body. Several
researchers have used antimicrobial finishes to provide fabrics with barriers against
microorganisms. With a view to develop antimicrobial textile materials by making
use of organic and inorganic compounds, antibiotics, heterocyclic and quaternary
ammonium compounds. Later organic stud the compounds like poly-halogenated
phenols, their esters and bisphenols also used to impart antimicrobial activity. Many
commercial antimicrobial agents are also available but their toxicity to people and
the environment are to be ascertained. An innovative approach to make the cloth
microbial resistant is to apply the plant extracts containing active substances. These
natural antimicrobial substances are not only eco-friendly but also from renewable
sources. Botanical research shows that some plant species exhibit antibacterial,
antiviral and antifungal properties. Essential oils extracted from various parts of
herbs such as the leaves, stems, flowers, fruits, roots, seeds, barks, etc. possess
antimicrobial activity. Some of the natural antibacterial products are neem oil, clove
oil, karanja oil, tulsi oil, cashew shell oil, henna or mehendi, drum stick tree, in this
project we are using clove oil as an antimicrobial finish.

36
In addition, the antimicrobial that is released to the environment may interfere with
other desirable microbes, such as those present in waste treatment facilities. The
second type of antimicrobial finish consists of molecules that are chemically bound
to fibre surfaces. These products can control only those microbes that are present on
the fibre surface, not in the surrounding environment. Bound' antimicrobials,
because of their attachment to the fibre, can potentially be abraded away or become
deactivated and lose long term durability.

Antimicrobial finishes that control the growth and spread of microbes are more
properly called biostats, i.e. bacteriostats, fungistats. Products that actually kill
microbes are biocides, i.e. bacteriocides, fungicides. This distinction is important
when dealing with governmental regulations, since biocides are strongly controlled.
Textiles with biostatic properties, however, are subject to fewer regulations.

The actual mechanisms by which antimicrobial finishes control microbial growth

are extremely varied, ranging from preventing cell reproduction, blocking of
enzymes, reaction with the cell membrane (for example with silver ions) to the
destruction of the cell walls and poisoning the cell from within. An understanding
of these mechanisms, although important for microbiologists, is not really a
requirement for the textile chemist who applies and evaluates the effectiveness of
antimicrobial finishes.

2.8 UV- PROTECTION FINISHING OF SILK

One of the major problems with silk fibres is their susceptibility to photochemical
degradation. UV-absorber organic compounds that absorb UV-radiation can impart
some protection by dissipating the energy as heat. UV absorbers are organic or
inorganic colourless compounds with a very strong absorption in the UV range of
290-360 nm. UV absorbers incorporated into the fibres convert electronic excitation

37
energy in to ther mal energy. They function as both radical and oxygen scavengers.
The high-energy shortwave ultraviolet radiation (UVR) excites the UV absorber to
absorb high energy. The absorbed energy may then be dissipated as longer-wave
radiation. Alternatively, isomerisation can occur, and the UV absorber may then
fragment in to non-absorbing isomers. Screening agents, energy transfer agents and
pigments can also be used to deactivate the fibre degradation process.

Organic UV absorbers are derivatives of o-hydroxyl benzophenones, o

hydroxyphenyltriazes and o-hydroxyphenyl hydrazines. The orthohydroxyl group in
the molecule helps in absorption and to make the compound soluble in alkaline
solution. Organic compounds like benzotriazole, hydrobenzophenone and
phenyltriazine can be used by normal padding or coating applications. Ortho
hydroxyphenyl and diphenyltriazine derivatives have excellent sublimation fastness
and a self-dispersing formulation. It can be applied by a pad thermosol process and
also in print pastes. The presence of organic pigments in the fibres helps in better
diffusion of light from the substrate, thus providing better protection. Titanium
dioxide and other ceramic materials have an absorption capacity in the UV region of
280-400 nm and reflects visible and IR rays.

38
 CHAPTER III
MATERIALS AND METHODOLOGY

3.1 INTRODUCTION

Almost every plant part produces bioactive compounds called phytochemicals.

These phytochemicals incorporate with other nutrients to form the guard
organization across the discrete number of diseases and environmental factors.
Amino acids, sugars, proteins, and chlorophyll are regarded as primary metabolites.
All other phytochemicals such as alkaloids, flavonoids, tannins, terpenoids,
saponins, and essential oils are classified under secondary metabolites. All the above
quoted compounds are synthesized in greater number of plants which are recognized
for their therapeutic activity. Syzgium aromaticum (clove) is the flower bud of a tree
associated to the genealogy of Myrtaceae. Clove is derived from the Latin ‘clavus’,
which means nail due to its resemblance with the shape. The clove tree is an
evergreen tropical plant which flowers twice every year. Cloves are the unopened
buds and harvested when the outer green leaves have changed from green to a yellow
pink.

The tree raises upto 8-12m tall, with broad leaves and crimson flowers associated
into bunch at terminal end. Cloves are frequently used as spices in different cuisines
such as Asian, African and Middle East, and observed as time-honored biologic
medication for local analgesics, anthelmintics (antiparasitic) and in dental treatment.
Topical administration of clove extract over the bowel region is used to cure
abdominal cramps. The preeminent component in clove extract is eugenol (70-90%).
The IUPAC name of eugenol is 4-allyl-2-methoxyphenol with molecular formula
C10H12O2. It has boiling point of 2540C. Eugenol combined with Zinc oxide is used
in filling dental cavities. Additional to eugenol, cloves contain acetyl eugenol,
betacaryophyllene, and vanillin.
39
Cloves also contain some nonvolatile substances such as tannins (10-13%),
tnterperen (2-4%), sterols and flavonoids Such as isobifiorin and bi-florin. Besides,
it also shows better antifungal property against Candida asperg and Dermatophyte
species.Though application of clove in the medical field is quite common, limited
study has been carried out to use clove extract as natural antimicrobial agent and as
a fragrance producing agent on textile substrate. Thus, it was thought worthwhile to
study and evaluate the antibacterial potency of phytochemical from Syzygium
aromaticum (clove) and fragrance intensity on silk woven fabric.

In this project the clove oil extracted from clove bud applied on silk fabrics to give
antimicrobial finish and their effects and durability is evaluated.

Materials:

1. Clove Extract

2. Bleached Silk Fabric

Preparation of materials:

Clove:

Clove flower buds (S aromaticum) purchased from the local market at Davangere,
Karnataka, India, and about 50 gm of cloves are measured and crushed using mortar
and pestle. The powdered cloves are stored in airtight container for future use.

This study has been carried out to use clove extract as natural antimicrobial agent on
textile substrates.

40
Fabric:

Bleached silk fabric to be used to carry out the study, purchased from the retail shop
at Davangere, Karnataka, India.

Solvent:

Dichloromethane can be used as a solvent (analytical grade) for purification of clove

bud.

3.2 GEOMETRICAL PARAMETERS OF SILK FABRIC:

1) EPI
2) PPI
3) Count of yarn.
4) Cover factor
5) Thickness.

Procedure:
Small pieces of silk fabrics are analyzed at Textile Testing Laboratory at Textile
Technology department for its geometrical parameters using counting glass, Beasley
Balance and thickness gauge.

Apparatus used:
1. Counting glass.
2. Beasley balance
3. Thickness gauge.

41
1) Threads/unit space:
 Counting glass is used to calculate the Ends Per Inch and Picks Per Inch in the
defined unit space.
1. Warp yarns- Ends.
2. Weft yarns - Picks.

 Take 5 Trails were taken and Average is calculated.

2) Count (Ne) and linear density (Tex):

Count (Ne) is calculated on Beasley balance using template to wrap the raveled yarn
from the particular fabric to be tested.
 Tex= 590.5/Ne.

3) Cover Factor:
Fabric is constructed by warp & weft yarns.
 Cover factor (K) = Threads per inch/√(counts) = n/√N
Warp cover factor = K₁
Weft cover factor = K2
 Hence Cloth cover factor Kc = K1 + K2 – K1K2/28

3) Fabric thickness:
Thickness gauge is used to measure the thickness of the fabric.

 Fabric weight: 10 x 10 cm Template

 Quadrant balance – gm/cm2

42
3.3 EXTRACTION OF ESSENTIAL OIL FROM CLOVE:

The components present in clove are said to be a volatile compound and extracted
using steam distillation. Initially, measure 10 gm of powdered cloves and add 80 ml
of distilled water (H20) to the round bottom flask of the steam distillation apparatus
and carry out distillation for 2 hours. Add about 20 ml of distilled H20 every 30
minutes to avoid burning of the sample. The distillate is collected and then subjected
to liquid detraction. Liquid-liquid extraction is performed by adding three portions
of 15 ml of dichloromethane (DCM) to 50 ml of distillate in the separate funnel as
per standard protocol. The collected extract is dehydrated using anhydrous sodium
sulfate. Then, deal the extract with rotary evaporator at 400 C to evaporate the
solvent, and concentrate the pure extract.

Clove flower buds about 50 gm are measured, crushed and powdered

Steam distillation

Clove powder + Distilled water for 2 hours

Add Dichloromethane
(Liquid-liquid extraction)
Collect the Clove extract

Dehydrate the collected Clove extract using anhydrous sodium sulfate

Evaporate the solvent with rotary evaporator at 400C

Collect the pure Clove extract

1. Flow chart for extraction of clove oil through distillation

43
3.4 TREATMENT OF SILK FABRIC WITH ALUM:

Alum is a representative name of potassium aluminum sulfate. Its appearance is

slight crystalline powder (Fig 1). Its chemical formula is KAI(SO4)2.12H20. It is a
good soluble salt in water and is used as adhesive which helps to bind finishing
material on the fabric surface. The bleached and mercerized silk fabric is treated
with 5% alum on weight of the material. This treatment was carried out in Chemical
processing laboratory at Textile Technology department. Weight of the material is
calculated on electronic balance (Fig 2). The material and liquor ratio should be 1:30.
The alum dissolves in water and kept in water bath (Fig 3) at temperature of 70°C.
The silk fabric is treated for about 1 hour and dried in hot air oven.

Bleached Silk fabric

Application of 5% alum (Potassium aluminum sulfate)

MLR 1:30 at 700C for 1 hour

2. Flow chart for treatment of silk fabric with alum

Figure 1: Alum Figure 2: Electronic Balance

44
 Figure: 3

3.5 APPLICATION OF CLOVE EXTRACT TO SILK FABRIC:

The alum treated samples are treated with purified extract of clove solution with
50% concentration for 1 hour at 500C in dye bath (Fig 3). This treatment was carried
out in Chemical processing laboratory at Textile Technology department. Then, the
samples are dried in hot air oven (Fig 4) at 100°C for 10 min. This treatment was
carried out in Chemical processing lab at Textile Technology department.

Alum treated Silk fabric samples

Application of purified Clove extract solution with 50% concentration for

1 hour at 500C
Drying
Hot air oven at 100 C for 10 minutes
0

45
3.6 MEDIUM OF PREPARATION FOR THE GROWTH OF BACTERIA:

Culture medium or growth medium is a liquid or gel designed to support the growth
of microorganisms. There are different types of media suitable for growing different
types of cells. After the preparation of the agar media, pour it in petri dish and keep
it in cold temperature undisturbed for 3 hours. Then, observe it for any contamination
of microorganisms.

Preparation of Inoculum:

Microbial Type Culture Collection (MTCC) strains of E. coli is prepared in

respective LB and Nutrient broth followed by required period of incubation of 24
hours and 48 hours respectively for further studies. The bacterial culture and nutrient
agar used for the study are shown in Fig 5 (a) and (b)

(a) (b)

Fig.5 (a) Bacterial culture and (b) Nutrient agar

46
Inoculation:

The culture method employed here is pour plate method. The inoculum is seeded at
the center of the Petri plate and the media was poured and rotated in clock wise
and anti-clock wise direction (Fig 6). The plate was allowed for solidification. All
the steps were carried under sterile conditions.

Figure 6: Inoculating

Evaluation of antimicrobial activity (qualitative agar diffusion test AATCC

147-2004)

Evaluation of antimicrobial activity was carried out for untreated and treated fabric
samples qualitatively by agar diffusion test AATCC 147-2004 (American
Association of Textile Chemists and Colorists). Bacterial cells were inoculated on
nutrient agar plates over which textile samples were laid for intimate contact. The
plates were then incubated at 37°C for 18-24 hours and examined for growth of
bacteria directly underneath the fabrics and immediately around the edges of the
fabrics (zone of inhibition). No bacterial growth directly underneath the fabric
sample indicates the presence of antimicrobial activity.

The inhibition zones were measured in terms of the distance in millimeters from
the edge of the disc to the zone edge. Measuring the zone size enable the strain to
47
be categorized as sensitive or resistant. The auto clave and Bacteriological
Incubator used are shown in the Fig 7 and 8

Fig. 7 Autoclave Fig. 8 Bacteriological Incubator

Bacteriological Incubator: Bacteriological incubator shown in the Fig 7 is similar

to that of oven. The temperature of the incubator is maintained at the desired level
by an automatic device called thermostat which acts as the connection off when the
temperature reaches the point for which the thermostat is set and turn on again
when the temperature falls slightly that point.

Laminar Air Flow Unit: The laminar air flow system is used for reducing the
danger of infection while working with pathogenic microbes is shown in Fig 9 The
laminar air flow works on the principle of application of fibrous filters in air
filtration. In this system, air of closed cabinet or room is made to pass through
High Efficiency Particulate

48
 Figure 9 Figure 10

Micropipette: A pipette is a laboratory tool used to transport a measured volume

of liquid. A pipette that dispense between 1-1000 µl are termed as Micropipette, is
shown in Fig 10

3.7 ANTIBACTERIAL ACTIVITY OF CLOVE EXTRACT ON SILK

FABRIC:

Agar well diffusion method is used to analyze antimicrobial properties of samples.

The medium can be prepared on petri dish. The bacteria used here is Escherichia
coli, the samples punched and a small circle of samples are placed in the medium
and keep undisturbed for 24 hours. The zone formation indicates the capability of
antimicrobial action of the clove solution on the silk fabric.

The bacteria swab uniformly across a culture plate. A filter paper disk, impregnated
with the compound to be tested, then place on the surface of the agar. In agar well
diffusion method, material required for process is kept under UV treatment for 15
min. After UV treatment, the petri plates are labelled properly and streaked with

49
bacterial strain using cotton swab from left to right and vice versa. Then, by using
well puncture, holes are created and the clove extract is injected. Add the clove
extract into the wells with concentration of 50%. Control it without adding eugenol
in the well. After certain fecundation period of 24 hours at 37°C, the zone of
inhibition is to be observed.

3.8 WASHING TEST

Washing test should be carried out according to standard ISO 105 C06 (modified)
using a detergent (Hostapal MRN liq C). This is free from alkyl phenyl ethoxylates
(APEO), Clove solution treated samples are immersed in washing bath for 30 min
at 400 C, rinsed with water and air dried. In such a way, 20 washing cycles to be
performed. After each 5 washing cycles, samples to be evaluated for antimicrobial
activity and fragrance intensity tests.

3.9 COLOR STRENGTH TEST

Clove solution treated samples are tested under reflectance spectrophotometer

(Gretag Macbeth with Des light using Colour control software interfaced with IBM
PC. The Chroma Flash Colour Matching System is used to evaluate K/S value for
comparison of colour between clove solution treated silk samples and washed
samples treated with different cycles. K/S is the colour value which is calculated
from Kubelka-Munk equation (1).

K/S= (1-R)2 /2Rx 100----------------- (1)

where,

K is absorption coefficient:

S is scattering coefficient:

50
R is spectral reflectance of the colored samples at a wavelength of maximum
absorption (where the reflection is minimum).

The percentage colour strength of the samples was determined using the formula:

Colour strength = (K/S sample + K/S standard) x 100------------- (2)

3.10 FRAGRANCE INTENSITY TEST

Fragrance applied fabric is to test the intensity of fragrance by subjective method.

The treated fabric tests the presence of fragrance for 5 weeks continuously. The
rating required is 5, 4, 3, 2, 1.0 about the intensity of fragrance in the fabric, with 5
to be the maximum and D' to be non-detectable fragrance.

51
 CHAPTER IV

RESULTS AND DISCUSSION

4.1 INTRODUCTION:
In this chapter, brief analysis of results obtained from various experiments conducted
in the chapter 2 is done and suitable discussion is made.

1. The clove oil was extracted from clove bud using steam distillation method.
2. Geometrical properties of bleached silk fabric are calculated and noted.
3. Bleached silk fabric was treated with potassium aluminum sulphate (Alum).
4. Antimicrobial finishing on silk was carried with clove extract.
5. Antimicrobial activity was estimated using agar well diffusion method.
6. Fragrance test was conducted on clove extract finished silk fabric.
7. Washing test was conducted to test the antimicrobial activity and fragrance
intensity.
8. Color strength treated sample were tested in reflectance spectrophotometer.

52
4.2 GEOMETRICAL PROPERTIES OF BLEACHED SILK FABRIC:

Sl.no Geometrical Properties Before Finish After Finish

1 EPI 100 100

2 PPI 96 96

3 Count of Yarn Warp=147.62 Warp= 147.62

Weft=111.41 Weft= 111.41
4 Cover Factor 14.65 14.65

6 Stiffness Warp (Avg) = 1.4 Warp (Avg) = 1.4

Weft (Avg) = 3.8 Weft (Avg) = 3.8

7 Crease Warp=105 Warp= 103

Weft=102 Weft= 100

Table 1: Geometrical properties of bleached silk fabric

53
4.3 THE ANTIMICROBIAL ACTIVITY OF FINISHED SILK FABRIC AND
THE ZONE OF INHIBITION ARE SHOWN BELOW:

1. Zone of inhibition for finished silk fabric

Sl.no Fabric Zone of inhibition (mm)
[E.coli]
1 Before wash 20
2 After wash 18
Table 2: Zone of inhibition for finished silk fabric

2. Antimicrobial activity of finished silk fabric

i) Before Finishing ii) After Finishing

Figure 11

54
3. Effect of washing on bacterial resistance, relative color strength, color change
and fragrance intensity
The extract of clove with distinct concentration of 50% was used to explore the
response of eugenol treated silk fabric on E. coli. The treated fabrics were added to
the prepared well in the agar plates. Zone of inhibition was measured and it showed
resistance towards eugenol.
With a view to understand the effect of washing cycle on antimicrobial activity of
clove solution treated samples, 4 different levels of washings i.e., 5,10,15 and 20,
were performed keeping unwashed sample 0 as control. As shown in table 2 and
figure 1, there in reduction in bacterial resistance due to increase in washing cycles.

Washing Zone of Reduction Relative Color Fragrance

cycle inhibition in bacterial color change intensity
(mm) resistance strength (Visual rating
(%) (%) gray scale
rating)
0 ( Clove 20 0 100 3.0 5.0
treated
sample
without
washing)
5 18 15 90 3.5 4.5
10 18 20 80 4.0 4.0
15 18 25 80 4.5 4.0
20 18 30 80 4.5 4.0
Table 2: Effect of washing on bacterial resistance, relative color strength, color change and
fragrance intensity

55
Antimicrobial activity of unwashed sample shows 30 mm zone of inhibition. With
increase in washing cycles from 5 to 20, there is reduction in bacterial resistance
from 7% to 30%.

Relative color strength of treated samples also exhibits a decreasing trend with the
increase in cycles of washing. Relative color strength of 80% was achieved after 20
washing cycles. Similar trend was noticed in the case of visual color change by gray
scale rating. After 20 washing cycles, color change rating of 4.5 is found in gray
scale assessment. It is corroborated from the results that, with the increase in washing
cycles, there is decrease in bacterial resistance and relative color strength. As
expected, color change rating of the treated samples increases because of removal of
color from the cotton fabric due to washing. Color change rating becomes constant
after 15 washing cycles. It has been observed that the intensity of fragrance in the
fabric slowly deteriorated with different cycles of washing used in the wash. At the
end of 20 wash cycles, the fragrance intensity was found to be 2 as the visual rating
using subjective method.

This indicates that fixation of clove solution with silk fabric is medium, and further
investigation is needed to explore the use of a suitable fixing agent in exhaust method
of application to improve the bacterial resistance and fixation of color for
commercial applications.

56
 CHAPTER V
CONCLUSION
From the results and discussion, the following conclusion may be drawn.
 The component responsible for antimicrobial property was extracted from clove
bud using standard procedure.
 The clove extracted from the clove bud was applied on silk bleached sample
using alum. The treated fabric shows the good antimicrobial property against E.
coli and this property is retained in the silk fabric after several washes.
 Fragrance of clove treated silk fabric was determined by subjective method. It
shows the fragrance rate ranging from 4-5.
 The relative color strength of finished silk fabric was evaluated. It has been
observed that antimicrobial activity, relative color strength and intensity of
fragrance of clove treated silk fabric retains its properties after repeated washing
cycles.

57
 SCOPE OF FUTURE WORK
1. Antimicrobial finish with clove extract is environmental friendly process and
health protecting, preventing diseases.
2. Antimicrobial finish with clove extract prevents the growth of bacterial and
microorganisms.
3. Antimicrobial finish with clove extract on the garments prevents unpleasant odors
due to bacterial growth or sweating.
4. Antimicrobial finish with clove extract on a fabric improves durability and
serviceability by promoting a safe and sanitary environment.
5. These type of finished fabrics can be used in a
 Health care textiles,
 Medical textiles,
 Undergarments,
 Costly Fabrics.

58
 REFERENCES

1. Ceylan, E. and Fung, D., 2020. Antimicrobial Activity of Spices.

2. L. A. Shelef., 2020. Antimicrobial Effects of Spices.
3. G. Telagavati et.al., 2020 Application of Prickly Chaff Leaves as Herbal
Antimicrobial Finish for Cotton Fabric Used in Healthcare Textiles.
4. Cui, H., Zhang, C., Li, C. and Lin, L., 2020. Antimicrobial Mechanism of
Clove Oil On Listeria Monocytogenes.
5. Murugesh Babu, K. and Ravindra, K., 2020. Bioactive Antimicrobial Agents
for Finishing of Textiles for Health Care Products.
6. http://www.academicjournals.org/ajmr.2020. Chemical Analysis and
Antimicrobial Activity of the Essential Oil of Syzigium Aromaticum (Clove).
https://www.researchgate.net/publication/237521165_Chemical_analysis_an
d_antimicrobial_activity_of_the_essential_oil_of_Syzigium_aromaticum_cl
ove
7. Gao, H., Yang, H. and Wang, C., 2017. Controllable preparation and
mechanism of nano-silver mediated by the microemulsion system of the
clove oil. Results in Physics, 7, pp.3130-3136.
8. L. B. Bullerman et., al, 2020. Inhibition of Growth and Aflatoxin Production
by Cinnamon and Clove Oils. Cinnamic Aldehyde and Eugenol.
9. Subrata Das. et., al Antimicrobial Activity and Fragrance of Clove Bud on
Woven Silk Fabrics .2020 Available at: Asian Dyer,October-November .2020
10. Schindler, Wolfgang & Hauser, Peter. (2004). Chemical Finishing of
Textile. 10.1533/9781845690373.74.
11. Asim Kumar Roy Choudhury,13 – (2017) Wool and silk finishing processes,
In Woodhead Publishing Series in Textiles, Principles of Textile Finishing,
Woodhead Publishing, Pages 417-466.

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