76 Y.

Y. Laoong-u-thai: The Evaluation of Eucaliptus Leaf Extract for Dyeing and Its Antibacterial Properties on Silk and Wool Fabrics

The Evaluation of Eucalyptus Leaf Extract

for Dyeing and Its Antibacterial Properties on
Silk and Wool Fabrics
Yanisa Laoong-u-thai1* and Rattanaphol Mongkholrattanasit2
1
Department of Chemical Engineering, Faculty of Engineering, Burapha University,
Chonburi, Thailand
2
Department of Textile Chemistry Technology, Faculty of Industrial Textile and Fashion Design,
Rajamangala University of Technology Phra Nakhon, Bangkok, Thailand

ABSTRACT but also in daily use. Recently, the antimicrobial

agent applied to fabrics have been synthetic or
In an effort to develop antibacterial fabrics, we natural substances such as formaldehyde, heavy metal
studied the properties of eucalyptus leaf extract for ions (silver, copper), antibiotics and natural dyes [1-
dyeing and antibacterial activity on natural textiles. 3]. There are many reports which showed the
Two types of natural fabric, silk and wool, were dyed antimicrobial activity of natural dyes beside their
with 20, 40 and 80% owf of the extract as natural dye coloring property including lawsone from henna,
using 40% owf of four different types of mordant juglone from walnut and lapachol from alkanet [4].
including Al, Cu, Fe and Sn. The dyed wool and silk Natural dyes have a wide range of shades that can
fabrics were shown a shade of yellowish-brown except be obtained from insects, minerals, fungi and various
when using Fe mordant which resulted in a shade of parts of plants including root, bark, leaves, flowers,
dark grayish-brown. Agar diffusion method was skins, fruits and shells of plants [5-6]. Natural dyes
performed for inhibition zone observation of dyed are well known for their uses in coloring of food
fabrics against Staphylococcus aureus and Escherichia substrate, leather, wood, as well as natural fibers
coli, gram positive and gram negative bacteria, as (wool, silk, cotton and flax) as major area of
common pathogen. The result showed that the best application since ancient time [7]. Some natural dyes
antibacterial property was observed from wool fabrics are not only dye with unique and elegant colours but
dyed with 20% owf dye and 40% owf Cu against both also provide antibacterial, deodorizing and UV
Staphylococcus aureus and Escherichia coli which protective functions to fabrics [8-14]. Nowadays, the
were shown 1.26 and 0.98 cm zone of inhibition. use of natural dyes in textile application has been
Meanwhile, both silk and wool fabrics which were witnessing a rapid growth. This is mainly attributable
dyed with 40% owf dye and 40% owf Cu, gave the to strict environmental standards set by many
high activity against Staphylococcus aureus with 0.88 countries/ organizations to avoid the health hazards
and 0.86 cm diameter of clear zone. In contrast, these associated with synthetic dyes used in textile. The
fabrics showed low activity against Escherichia coli. recent ban on the use of azo dyes by European Union
This result clearly showed that all dyed fabrics using has also increased the scope for the use of natural
Cu as mordant showed the antibacterial activity both dyes [15].
gram positive and negative strains while other Eucalyptus is one of the most important source of
mordants showed less or no activity. natural dye that gives yellowish-brown colorants. The
coloring substance of eucalyptus has ample natural
Keywords: Antimicrobial fabric, Eucalyptus extract, tannins and polyphenols varying from 10% to 12%
Antibacterial activity, Natural fabric Natural dye [16]. The major coloring component of eucalyptus
bark is quercetin as also an antioxidant. It has been
1. INTRODUCTION used as a food dye with high antioxidant properties
[17]. Eucalyptus leaves contain up to 11% of the
Medical and hygienic textiles in terms of major component of tannins (gallic and ellagic acids)
antibacterial fabrics have become important areas in whil minor substances are flavonoid (quercetin, and
the textile production not only in medical applications rutin, etc.) [18-20]. Tannins and flavonoids are
considered very useful substances during the dyeing
* Corresponding author: E-mail: [email protected] process because of their abilities to fix dyes within
fabrics. Dyeing process using natural dyes, normally
requires “mordants”, which are metallic salts of
aluminum (Al), iron (Fe), chromium (Cr), copper
(Cu), among others, for ensuring a reasonable fastness
KMITL SCIENCE AND TECHNOLOGY JOURNAL VOL.13, NO.2, 2013 77

of the color to sunlight and washing [21]. The metal leaf extract dye solutions versus absorbance at the
ions of these mordants take action as electron wavelength mentioned.
acceptors for electron donors to form coordination
bonds with the dye molecules, resulting them E. Dyeing method
insoluble in water [22]. Pre-mordanting dyeing was used. The dye
In this study, the dyeing, fastness and concentration was varied at 20%, 40% and 80 % on
antibacterial properties of silk and wool fabrics using weight of fabric (% owf.) of eucalyptus leaf extract
an aqueous extract of eucalyptus leaves as natural dye dye, and four types of mordants (Al, Cu, Fe, and Sn)
were investigated. Different factors affecting dyeing were used with 40% owf. of each concentration of dye.
ability were also thoroughly examined. The pH of the dyeing solution was adjusted to 4 with
an acetic acid solution. The silk and wool fabrics were
2. METERIAL AND METHODS dyed at 90°C at a liquor ratio of 1:50 for 60 minutes.
After dyeing, the dyed samples were rinsed
A. Fabrics with cold water, washed in a bath with a liquor ratio
of 1:50 using 1 g/l of the soaping agent, Syntapon
A commercially produced wool fabric (plain- ABA, at 80°C for 5 minutes, then rinsed and finally
weave) was scoured with an aqueous nonionic air-dried at room temperature.
surfactant solution at a temperature of 45°C for 30
minutes, then it was thoroughly rinsed, and air dried F. Evaluation of colour strength and fastness
at room temperature. The scoured and bleached silk properties
fabric (plain-weave) used thoughout this study, was The colour strength (K/S) and CIELAB of the
supplied by Chul Thai Silk Co., Thailand. dyed samples were evaluated using a
spectrophotometer (Datacolor 3890). All measured
B. Mordants and Chemicals samples showed a maximum absorption wavelength
The following laboratory-grade mordants were (λmax) value at 400 nm. The K/S is a function of
used: aluminium potassium sulfate dodecahydrate colour depth and is calculated by the Kubelka-Munk
(AlK(SO4)2.12H2O), ferrous (II) sulfate heptahydrate equation, K/S = (1-R)2/2R, where R is reflectance, K
(FeSO4.7H2O), copper (II) sulfate pentahydrate (Cu is the sorption coefficient, and S is the scattering
SO4.5H2O), and stannous chloride pentahydrate coefficient.
(SnCl2.5H2O). An anionic wetting agent, Altaran S8
(sodium alkylsulfate), and the soaping agent, G. Bacteria and culture medium
Syntapon ABA, were supplied by Chemotex Děčin, Bacterial strains that were used in this research
Czech Republic. including Staphylococcus aureus and Escherichia coli,
were supported by Department of Microbiology,
C. Instrument Prince of Songkla University, Thailand. The
The mordanting and dyeing were carried out in a propagation of bacteria and anti-bacterial assay were
dyeing machine (Linitest Type 7421) with undergoing using nutrient medium.
programmable time and temperature control. A
Datacolor 3890 was employed for the colour strength H. Propagation of bacterial strains
measurements. Nutrient broth and agar were prepared in the
usual fashion [24]. Bacterial glycerol stocks of
D. Dye extraction from eucalyptus leaves Staphylococcus aureus and Escherichia coli, were
Fresh eucalyptus leaves (Eucalyptus streaked on nutrient agar and incubated at 37°C
camaldulensis) were dried in sunlight for one month overnight to get single colony, which will be used for
and crumbled using a blender; and then they were antibacterial test in next procedure.
used as the raw material for dye extraction, which
was achieved by a reflux technique. Seventy grams of I. Antibacterial fabrics assay using agar diffusion
the crumbled eucalyptus leaves was mixed with 1 liter method
of distilled water and refluxed for 1 hour. The sample
was then filtered and the dye solution was separated The disk-agar method was performed for
into two portions, one for evaporating under reduced antibacterial activity. Natural fabrics, wool and silk,
pressure (rotary evaporator), and another one for which were use for antibacterial test, were dyed with
dyeing. A crude dye extract of eucalyptus leaves eucalyptus dye at two different concentration of 20%
obtained from the rotary evaporator was crumbled owf and 40% owf. Each dyed fabric was applied with
with a blender and used for obtaining a standard various mordant types of Al, Cu, Fe and Sn at the
calibration curve. A dilution of the eucalyptus leaf 40% owf mordant concentration. The 3-4 bacterial
extract gives a relatively clear solution with a linear colonies were seeded in 10 ml nutrient broth and
dependance on concentration–absorbance relation at incubated at 37°C overnight. The 500 µl of culture
an absorption peak (λmax) of 262 nm [23]. The was spread over the surface of nutrient agar. Five
concentration of 20 g/L was calculated from a mm-diameter fabrics were cut and bored in the agar
standard curve of concentrations of the eucalyptus plates. Antibacterial activity was evaluated after
incubation at 37°C overnight by measuring diameters
78 Y. Laoong-u-thai: The Evaluation of Eucaliptus Leaf Extract for Dyeing and Its Antibacterial Properties on Silk and Wool Fabrics

of inhibition zone. Original wool and silk were served dye concentration. The mordant activity sequence for
as negative controls. Each test was performed the silk was FeSO4 > CuSO4 > AlK(SO4)2 > SnCl2 >
triplicate for statistic evaluation. without mordanted and FeSO4 > CuSO4 > SnCl2 >
AlK(SO4)2 > without mordanted for the wool fabrics.
3. RESULTS AND DISCUSSION In all cases, the ferrous sulfate mordant yielded the
best dyeing results. The wool fabric dyed with
A. Effect of dyeing conditions eucalyptus leaf extract showed a higher colour
strength than the silk fabric, which is because the
Table 1 shows the colour values of the silk and wool fabric contains more functional groups than the
wool fabrics dyed with eucalyptus leaf extract dye silk fabric [25].
solution. The K/S values increase with an increase of

TABLE I. COLOUR VALUE OF SILK AND WOOL FABRICS DYED WITH EUCALYPTUS LEAF EXTRACT BY PRE-MORDANTING AND USING 40 % OWF. OF
METAL MORDANTS AT DIFFERENT CONCENTRATION OF THE DYE

Ferrous sulfate and copper sulfate mordants are The obtained value results obtained (Table 1)
well known for their ability to form coordination show that wool and silk fabrics dyed without mordant
complexes and to readily chelate with the dye. As the and with stannous show yellowish-brown and bright
coordination numbers of ferrous sulfate and copper yellow colour, respectively. The samples mordanted
sulfate are 6 and 4, respectively, some coordination with alum and copper sulfate produced medium to
sites remain unoccupied when they interact with the dark yellowish-brown. With ferrous sulfate, the colour
fiber. Functional groups such as amino and carboxylic shade was darker and duller. This may be associated
acid on the fiber can occupy these sites. Thus, the with the change of ferrous sulfate into a ferric form by
metal can form a ternary complex on which one site reacting with oxygen in the air. Ferrous and ferric
with the fiber and the other site is with the dye [26]. forms coexist on the fiber and their spectra overlap,
Stannous and alum ions form weak coordination which results in a shift of λmax and thus
complexes with dye; they tend to form quite strong consequently a colour change to a darker shade [27].
bonds with the dye but not with the fiber, so they Additionally, the tannins in the eucalyptus leaf
block the dye and reduce the dye interaction with the extract combine with ferrous salts to form complexes,
fiber [26]. which also result in a darker shade of fabric [28].
KMITL SCIENCE AND TECHNOLOGY JOURNAL VOL.13, NO.2, 2013 79

B. Antibacterial property of dyed fabrics Benali, 2012 [29]. They reported that the essential oil
of the eucalyptus leaves has antimicrobia activity
Natural fabrics including wool and silk were dyed against gram-negative bacteria (E. coli) as well as
as mention earlier. Its antibacterial activity was gram positive bacteria (S. aureus). However, the
studied using agar diffusion method. The result structure of fabric can also effect the staining
showed that only wool which was dyed with 20% owf property of dye [30]. The molecular structure of wool
and 40% owf of eucalyptus dye concentrations, has much more amino acids with polar side residues
displayed the antibacterial activity (Table 2) while than silk. It affects the dye and heavy metal
silk that was dyed at the same condition, had no adsorption capacity of the fabric [31] resulting in
activity. This result was supported by Bachir and improved antimicrobial properties.

TABLE II. THE INHIBITION ZONE OF NATURAL DYE FROM EUCALYPTUS LEAVES COMBINED WITH METAL MORDANT TYPES FOR NATURAL FABRICS
STAINED, SILK AND WOOL

Moreover, the antimicrobial activity of dyed and silk on both positive and negative strains. The
fabrics combined with metal mordants was observed explanation likely lies in the increase of charge on the
(Table 2). The biggest clear zone was exhibited fabric due to the adsorption property of the metal
against both S. aureus and E. coli in the case of dyed ions (mordant) on the polar amino acids (fabric) [31]
wool with 20% owf of dye and 40% owf Cu mordant. cause the interaction between bacterial cell and fabric
It showed the inhibition zone of 1.26 and 0.98 cm, through Cu [32]. Cu is known as biocide [33] and
respectively. Meanwhile, other applied fabrics were effective against a wide range of gram-positive and
presented the antibacterial properties such as wool gram-negative bacteria including Bacillus subtilis,
and silk dyed with 40% owf dye and 40% owf Cu Pseudomonas aeruginosa, Escherichia coli and
mordant, showed the inhibitory zone of 0.88, 0.86 and Staphylococcus aureus [34-35]. The effect of Cu on E.
0.20, 0.48 cm against S. aureus and E. coli (Fig. 1). coli cell has also been studied [36]. It describes that
The suppression activity was also observed from wool Cu ion induces the formation of superoxide anions
dyed with 20% owf dye and 40% owf Fe mordant and hydrogen peroxide and cause DNA damage
against S. aureus and E. coli with the clear zone of resulting in cell death (Fig. 2).
0.16 and 0.22 and slight antimicrobial activity was
shown with Al and Sn mordants as well (Table 2).
This result reveals that mordant types especially Cu
enhanced the antibacterial property of the dyed wool
80 Y. Laoong-u-thai: The Evaluation of Eucaliptus Leaf Extract for Dyeing and Its Antibacterial Properties on Silk and Wool Fabrics

4. CONCLUSION Figure 1 Anti-bacterial activity of wool and silk fabrics

with 40% owf dye and 40% owf mordant types including Fe
The wool fabric dyed with eucalyptus leaf extract (1), Al (2), Cu (3) and Sn (4) with negative control of no
shows higher K/S values than the silk fabric. The use mordant (5). (A) Wool fabrics dyed with 40% dye and 40%
of a ferrous sulfate mordant gives rise to the best owf of mordants on E.coli culture and (B) on S.aureus
dyeing and exhibits a darker shade. The silk and wool culture. (C) Silk fabrics dyed with 40% owf dye and 40%
fabrics dyed with a eucalyptus leaf extract dye owf mordants on E.coli culture and (D) on S.aureus
solution with or without mordants showed a culture.
yellowish-brown shade (except with stannous chloride,
which produced the shades of bright yellow). The use
of mordants not only improves colour strength, but
also provides shade differences.
However, the study of antibacterial activity of
these dyed fabrics showed that dye concentration and
mordant typed had an effect on antimicrobial
property. The combination of dye and mordant typed
enhanced the bacterial inhibition of both positive and
negative strains. The best condition of antimicrobial
fabric was shown on dyed fabric with 40%owf dye
and 40%owf Cu mordant giving a 0.8 cm diameter of Figure 2 The effect of Cu on E. coli cell [36]
clear zone. However, other mordants also showed
antimicrobial activities: e.g. Fe of 20%owf and
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