1. Malaya Journal of Biosciences 2014, 1(1):19–23
ISSN:
Development of mosquito repellent using V. negundo
19
RESEARCH ARTICLE
Open Access Full Text Article
Development of Mosquito Repellent Fabrics using
Vitex negundo loaded Nanoparticles
Rajendran R1, Radhai R1 and Rajalakshmi V1*
1
PG and Research Department of Microbiology, PSG College of Arts and Science, Coimbatore-641014, Tamil Nadu.
*
For correspondence e-mail: raji.ajjii@gmail.com.
Article Info: Received 10 Nov 2013; Revised: 18 Nov 2013; Accepted 03 Dec 2013
ABSTRACT
Mosquito repellent textiles are one of the revolutionary ways to advance the textile field by providing the
much-needed features of driving away mosquitoes, especially in the tropical areas. The current study is
mainly carried out for the development of mosquito repellent fabrics using nanoparticle loaded with V.
negundo leaf extract. The nanoparticle loaded with V. negundo leaf extract were synthesized using ionic
gellification method. The synthesized nanoparticles were characterized using Dynamic Light Scattering
(DLS) and Fourier Transform Infrared Spectroscopy (FTIR) analysis. Repellent textiles were achieved by
padding cotton fabrics with nanoparticle loaded with V. negundo leaf extract using a conventional pad-dry
method. The finished fabrics were further analyzed for their mosquito repellent activity using Mosquito
Repellency Behavioral test. The results of the repellent activity are based on the test for evaluating the
effectiveness of mosquito repellent finishes. The treated fabrics showed 100% mosquito repellent efficiency.
The nanoparticles loaded with V. negundo leaf extract treated fabrics retained their activity until 15 washes.
These types of textiles protect the human beings from the bite of mosquitoes and there by promising safety
from the mosquito borne disease include malaria and dengue fever.
Keywords: Vitex negundo, mosquito repellent, DLS, FTIR, malaria, dengue.
1. INTRODUCTION
The population explosion and the environmental
pollution in the recent years forced the researchers to
find new health and hygiene related products for the
well-being of mankind. As the consumers are now
increasingly aware of the hygienic life style, there
is a necessity and expectation of a wide range
of economical and hygienic textile products. One of
the recent trends in textile industry is
‘nanotechnology’ which can provide high
durability for fabrics as they have a large surface
area to volume ratio and high surface energy,
thus presenting better affinity for fabrics and
leading to an increase in durability of the
function. The use of medicinal plants as a source of
relief from illness can be traced back to over five
million years in the early civilization of China, India,
and North east, which is as old as mankind. It has
been estimated that in developed countries such as
US, plant based drugs constitute about 25% of the
total drugs, while in fast developing countries such as
China and India, the contribution is about 80%. Thus,
the economic importance of medicinal plants is much
more to developing countries than to rest of the
world.
Vitex negundo is an herbal plant which is
available in most part of the world. Every part of this
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Development of mosquito repellent using V. negundo
plant is useful in many ways such as leaves are anti-
inflammatory [3,4] antihelmintic and anti-insecticidal
[5]. Roots are tonic, anti-rheumatic, wounds and
ulcers [2].
Mosquitoes are major vector in the transmission
of viral disease and parasite disease such as Yellow
fever, Japanese encephalitis, Malaria etc., the present
study mainly focuses on the development of
mosquito repellent cotton fabrics using nanoparticle
loaded with V. negundo leaf extracts. This research
also deals with the effect of treated cotton fabrics
against mosquitoes and to enhance the laundering
durability of the fabrics.
2. MATERIALS AND METHODS
2.1. Collection, Processing and Extraction of
V. negundo
The medicinal herb selected for the present study was
V. negundo which was collected in and around
Coimbatore. The collected leaves were shade dried at
room temperature to reduce the moisture content. The
leaves were then powdered and sieved. 20 grams of
the ground herbal powder was suspended in 100ml
ethanol and incubated overnight. The supernatant was
filtered twice using Whatman No.1 filterpaper and
the filtrate was used for further analysis.
2.2. Synthesis of nanoparticle loaded with
V. negundo leaf extract
Alginate nanoparticles were prepared by the
principle involving cation induced controlled
gelification of alginate [8]. Nanoparticle loaded with
V. negundo leaf extract was synthesized. About 5.0ml
of calcium chloride (18 mM) was added to 95.0ml of
sodium alginate solution (0.06%, w/v) containing
herbal extract. About 25ml of Chitosan solution
(0.05%, w/v) was added followed by stirring the
mixture in magnetic stirrer for 30 min and the
mixture was kept at room temperature overnight. All
the samples were then centrifuged at 1500 rpm for 15
min to remove any large aggregates. Centrifugation
under these conditions allowed the aggregates to
form pellet, leaving nanoparticles suspended in the
supernatant. The particle suspension was then
centrifuged at 8000 rpm for 15min at 25o
C to
separate free polymers from nanoparticles [1]. The
pellet containing the leaf extract loaded alginate
nanoparticles was washed five times and then
suspended in distilled water and stored at 4o
C for
further analysis. Alginate nanoparticle controls were
also prepared by the same procedure without loading
herbal extract.
2.3. Characterization of Nanoparticle loaded with
V. negundo leaf extract
The Nanoparticle loaded with V.negundo leaf
extract was characterized by physically using DLS
and chemically using FTIR analysis.
2.3.1. Dynamic Light Scattering analysis
The average nanoparticle size and zeta potential of
nanoparticles loaded with V. negundo extract were
determined by Dynamic light scattering was done
using Malvern zetasizer version 2.2. [8].
2.3.2. Fourier Transform Infrared Spectroscopy
(FTIR) analysis
FT-IR-spectra were obtained using a FT-IR
Spectrometer (Bruker tensor 27, Germany). Samples
obtained by centrifugation were used for FTIR
characterization. In this study, nanoparticle loaded
with herbal extract, Alginate chitosan
nanoparticles(control) and bulk herbal extract were
subjected to FTIR characterization in the scanning
range of 4000- 400 cm-1
. There were 100 scans for
each sample with a resolution of 4 cm-1
. No
smoothing functions and baseline correlation were
performed.
2.4. Finishing of cotton fabrics
Adsorbent sterile cotton fabric was purchased and
was then subjected to desizing. The desized fabric
was primarily coated with citric acid to ensure better
binding of the prepared formulation using pad-dry-
cure method. About 1 gm of the fabric 20 ml of the
nanoparticles and about 1.6 Gms of citric acid was
used as binder, the fabric was kept immersed in the
treatment solution for 20 minutes. The fabric was
then passed through a padding mangle (R. B.
Electronic and Engineering, Mumbai), running at a
speed of 15m/min with a pressure of 2 kgf/cm2
to
remove excess solution. A 100% wet pick-up was
maintained for all of the treatments. After padding,
the fabric was air-dried and then cured for 3 min at
1400
C and immersed for 5 min in 2 g/l of sodium
lauryl sulfate to remove unbound solutions and rinsed
to remove the soap solution followed by air-drying.
[13].
2.5. Assessment of Insect repellent property
The mosquito repellency efficiency of the V.
negundo loaded nanoparticle treated fabrics using
Mosquito Repellency Behavioral test.
2.5.1. Collection of mosquitoes
Anopheles mosquitoes were identified and
collected based on morphologic keys during the

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Development of mosquito repellent using V. negundo
evening hours. Mosquitoes were deprived of all
nutrition and water for a minimum of 4 hours before
exposure.
2.5.2. Mosquito Repellency Behavioral test
Specially designed two excito repellency test
chambers were used to evaluate the efficiency of
repellency activity. Laboratory tests were performed
during daylight hours only and each test was
replicated four times. Observations were taken at
one-minute interval for 30 minutes. After each test
was completed, the number of escaped specimens
and those remaining inside the chamber was recorded
separately for each exposure chamber, external
holding cage, and paired control chamber [14].
Efficiency of Mosquito Repellency (%) =
No. of specimen escaped + No. of specimen dead X 100
No. of specimen exposed
2.6. Wash durability analysis
Washing was carried out as per test no: 1 of IS:
687-1979. By using a neutral soap (5 gpl) at 40
±2°C for 30 min, keeping the material: liquor
ratio at 1:50, followed by rinsing, washing and
drying. After drying, the test samples were assessed
for antimicrobial activity using AATCC 100
procedure up to 30 laundering cycles.
3. RESULTS AND DISCUSSION
3.1. Characterization of Nanoparticle loaded with V.
negundo leaf Extract
3.1.1 Dynamic Light Scattering
The Average particle size and zeta potential were
analyzed using Dynamic Light Scattering. The
average particle size and zeta potential of the
nanoparticle loaded with V. negundo leaf extract were
found to be 182.6 nm and -30.3 mV respectively and
shown in Fig 1 & 2. According to Taraneh Gazori [7]
the stability of many colloidal systems is directly
related to the magnitude of their zeta potential. In
general, if the value of the particle zeta potential is
large, the colloidal system will be stable. On the other
hand, if the particle zeta potential is relatively small,
the colloidal system will agglomerate.
3.1.2. FT-IR analysis
FTIR was adopted to characterize the potential
interactions in the nanoparticles. FTIR spectra of
nanoparticle loaded with V. negundo leaf extract and
ethanol V. negundo extract were analyzed and shown
in Figure.2. In the spectra of herb loaded
nanoparticles, the broad band at 3440 cm-1
corresponded to the alcohols and the phenols; the
peaks at 3380 cm-1
was caused by N-H stretching
which showed the presence of primary and secondary
amines. The peaks at 3329 cm-1
and 3228 cm-1
belonged to the N-H stretching, this indicated the
presence of amides.
Figure 1. Average particle size -182.6nm using Malvern zetasizer
version 2.2.
Figure 2. Zeta potential (-30.3 mV) using Malvern zetasizer
version 2.2.
The O-H stretch of the spectra represented the
presence of carboxylic acids and the -CH2-OH stretch
represented hydroxyl groups [10]. The bands around
1130 cm-1
(C-O-C) stretching presenting in the IR
Spectrum of sodium alginate were attributed to its
saccharide structure of sodium alginate. In addition,
the bands at 1294 cm-1
were assigned to aromatic
amines [10]. In the, FT-IR spectra of V. negundo
loaded alginate chitosan nanoparticles, we can
observe the asymmetrical stretching of N-O groups
shifted to 1913 cm-1
. In addition, the absorption band
at 1151 cm-1
of chitosan shifted to 1515 cm-1
at after
reaction with alginate. The stretching vibration of -
OH at 3297 cm-1
shifted to 3380 cm-1
and became
narrow. Crude extract displayed peak characteristics
of the N-H stretching vibration at 3375 cm-1
and a

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Development of mosquito repellent using V. negundo
band with main peak at 3479 cm-1
indicative of the
O-H stretch of the alcohols and H-bonded phenols.
The characteristic absorption band of crude extract
appeared in the V. negundo loaded chitosan/ alginate
nanoparticles, which probably indicated that herb was
loaded inside the nanoparticles. These results
indicated the carboxylic groups of chitosan through
electrostatic interaction to from the polectrolyte
complex.
Figure 3. FT-IR spectra for the Leaf extract and V. negundo loaded
nanoparticles using Bruker tensor 27 spectrophometer scanned in
the range of 4000- 400 cm-1
3.2. Assessment of Insect repellent property
The mosquito repellent property of the
nanoparticle loaded with V. negundo leaf extract
treated fabrics using Mosquito Repellency Behavioral
test. The nanoparticle loaded with V. negundo leaf
extract treated fabrics showed 76% efficiency of
mosquito repellency and shown in Table.1.
Table 1. Assessment of mosquito repellent efficiency
S.No Fabric Treatments Efficiency of Mosquito
Repellency (%)
1. Ethanolic extract of 78
V. negundo leaves
2. Nanoparticle loaded with 76
V. negundo leaf extract
3.3. Wash Durability
The laundering cycles of treated fabrics were
analyzed using Wash durability test. The nanoparticle
loaded with V. negundo leaf extract treated fabrics
sustained mosquito repellency activity until 15
washes. While the fabrics treated with crude V.
negundo extract treated fabrics possesses less
efficiency than nanoparticle loaded with V. negundo
leaf extract treated fabrics due to the uniform coating,
better affinity and sustained release of the
nanoparticles (Table.2.). But in case of crude extract
treated fabrics does not possess sustained release of
the ant insecticidal compound like that of
nanoparticles. Rajendran et al., [11 &12] reported
that the higher laundering durability of the
nanoparticles was due to the smaller particle size,
uniform coating and controlled release of the
nanoparticles. i.e., herb loaded inside the particle
coated with chitosan which stay in the fabrics for
long time after 15 washing [7 & 11].
Table 2. Laundering cycles of treated fabrics using Wash
Durability analysis
S.No. Fabric Treatments No. of laundering Efficiency of
cycles mosquito
Repellency (%)
1. V. negundo leaf extracts 5 68
treated cotton fabrics 10 52
15 26
20 18
2. Nanoparticle loaded with 5 76
V.negundo leaf extract 10 72
treated cotton fabrics 15 70
20 64
4. CONCLUSION
Vector borne diseases are one the major problem in
developing countries. To avoid such kind of disease
transmission to humans, these kinds of fabrics can be
used. Using these fabrics, an efficient mosquito
repellent curtain, tents, bed nets etc., can be produced.
Therefore we suggest the finished fabric provide
evidence to be effective, economical and ecofriendly.
Conflict of Interest
The authors declare that they have no conflicts of
interest
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