Mosquito Repellent Property of Ylang Ylang
Mosquito Repellent Property of Ylang Ylang
Mosquito Repellent Property of Ylang Ylang
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Philippine Journal of Science Reyes et al.: Mosquito Repellent
Vol. 149 No. 4, December 2020 Property of “Ylang-Ylang”
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Figure 1. Photograph of ylang-ylang showing the leaves, flower, and seeds from a mature branch.
containing ylang-ylang essential oil extract using urea- and kept at room temperature until microencapsulation.
formaldehyde encapsulation compared with untreated
fabric, and 2) determine the mosquito repellency of cotton
Preparation of Urea Formaldehyde Polymer
fabric treated with different concentration of ylang-ylang
Fifty grams (50 g) of urea were mixed with 100 mL of 37%
essential oils.
formaldehyde. Thirty (30) drops of concentrated sulfuric
acid were added to the mixture, which was stirred constantly.
The mixture was centrifuged at 4,200 x g in batches and
the precipitates were pooled. The pooled precipitate was
MATERIALS AND METHODS resuspended and washed with distilled water, centrifuged
The study was approved by the Pamantasan ng Lungsod at 4,200 x g, and cleared of the supernatant. This procedure
ng Maynila’s (PLM) College of Medicine Publication and was repeated three times. The precipitate was filtered,
Review Committee and was registered to the Research collected, dried, and stored in an amber-colored bottle at 4
Grants and Administration Office, University of the °C until further use. All chemicals used in this study were
Philippines Manila. This study used a quasi-experimental laboratory-grade and sourced from Sigma-Aldrich (Merck
design and was conducted at the Biochemistry Laboratory KgaA, Darmstadt, Germany).
of PLM’s College of Medicine.
Microencapsulation Method
Sampling Method A simple coacervation method of microencapsulation
The cotton fabrics were purchased from a local fabric was done using a urea-formaldehyde polymer as the
shop in Manila and were woven with 100% cotton fabric coating material and ylang-ylang essential oil as the core
with 150 thread count, while the ylang-ylang essential material. Twenty grams (20 g) of urea-formaldehyde was
oil was purchased from a shop in Manila. The cotton mixed with 200 mL distilled water and left to swell for
fabrics, measuring 58 cm2 for each sample and weighing 30 min. One hundred milliliters (100 mL) of hot water
1.20 g each, were washed and bleached using sodium was added to the mixture while the mixture was being
hypochlorite (Zonrox™ Bleach, Greencross Inc., Manila, stirred constantly in a hot plate maintaining a temperature
Philippines) for an antimicrobial finish prior to the of 40–50 °C. The stirring was continued until all urea-
microencapsulation method. The fabrics were air-dried formaldehyde resins were dissolved. Ten milliliters (10
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Vol. 149 No. 4, December 2020 Property of “Ylang-Ylang”
mL) of ylang-ylang essential oil was added in the solution aegypti and reared at the laboratory condition of about 27 ±
and was stirred for 15 min. The drop-by-drop addition of 2 °C and 70 ± 10% humidity. Batches of about 3–5-day-old
20% sodium sulfate solution was done until the mixture mated but non-blood-fed mosquitoes were used in the test.
reached a volume of 250 mL in a span of 10 min with a The cone test was done in the laboratory condition similar
ylang-ylang concentration of 4%. The cotton fabric was to the rearing condition of the mosquitoes. The cone used
immersed in the microcapsule solution for 30 min. The was made of plastic with a base diameter of 12 cm and
fabric was squeezed and dried in an oven at 80–85 °C to a height of 6 cm. The untreated and treated fabrics were
cure the polymer and to ensure that excess formaldehyde placed at the base of the cone. Ten (10) susceptible, non-
in the fabric has evaporated. The fabric was kept in a blood fed female mature adult mosquitoes were released
closed amber glass container until further use. inside the cone. The repellency of adult mosquitoes was
observed, noted, and recorded at a 10-min interval for
Another set-up was done using a similar microencapsulation 1 h. Repellency in this study is defined as the quality
method but, instead of 10 mL, 2.5 mL of ylang-ylang or capacity to drive away and was measured using the
essential oil was used with a concentration of 1%. The number of landing/resting of the adult mosquitoes on the
third fabric sample was the untreated cotton fabric, which filter paper, and the % repellency value was calculated for
was only washed, breached and dried, and kept in a similar each time interval. The % repellency of adult mosquitoes
manner until further use. was computed using the formula:
% repellency = 100 – {X/N x 100}
Characterization of Urea-Formaldehyde
Microencapsulated Fabric where X is the number of mosquitoes landed/rested on
The morphology and size of different concentrations the filter paper and N is the total number of mosquitoes.
of microcapsules attached to the cotton fabrics were
examined using a scanning electron microscope (SEM). The mean % repellency ± standard deviation (SD) was
The SEM used was a JEOL 5310 model (JEOL Ltd., reported for the untreated and treated fabric with 2.5 mL
Tokyo, Japan) equipped with secondary and backscattered and 10 mL of ylang-ylang essential oil microencapsulated
electron detectors. It has a suitable accelerating voltage with the urea-formaldehyde polymer.
and magnification equipped with an AMETEK EDAX
ELEMENT energy dispersive spectroscopy system. Data Analysis
All samples were coated with gold to make the sample Data gathered from scanning electron microscopy were
conductive using a JEOL JFC-1200 fine coater (JEOL described and results of the % repellency in mean ±
Ltd., Tokyo, Japan) before setting it into the equipment SD were analyzed using one-way ANOVA. A post hoc
for visualization using an accelerating voltage under analysis was made using Dunnett’s multiple comparison
thermal vacuum. SEM micrographs were taken at an t-test. All statistical tests were conducted using the IBM
accelerating voltage of 15 kV. Micrographs were taken SPSS Statistics software. Statistical significance was set
with the SemAfore software (Insinooritoimisto Rimppi at p < 0.05.
Oy, Finland).
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Figure 2. Scanning electron micrographs of microcapsules taken at magnifications (A) 300x; (B) 1000x; and (C)
5400x with accelerating voltage of 15 kV. A black arrow is pointing to a microcapsule on the cotton fiber.
Mosquito Repellency Test fabric remained stable over the exposure period. Both mean
The mosquito repellency response to 10 mL of C. odorata % repellency of the treated cotton fabric was significantly
essential oil of UFY microcapsule-treated cotton fabric higher than the untreated cotton fabric (p < 0.05).
was observed at an increasing rate from 80 ± 10% to
88 ± 8.37% within 60 min of the exposure period. The
peak of mean % repellency was observed at 40–60 min
of exposure period from 86 ± 5.48% to 88 ± 8.37% (see DISCUSSION
Figure 2). The mean % repellency of 10 mL of C. odorata
Plants produce significant amounts of phytogenic volatile
essential oil of UFY microcapsule-treated cotton fabric
organic compounds, which are composed of terpenoids,
was significantly higher than the mean % repellency of
phenylpropanoids, benzenoids, and volatile fatty acid
2.5 mL of C. odorata essential oil of UFY microcapsule-
derivatives derived from various and different biosynthetic
treated cotton fabric at all intervals (p < 0.05).
pathways in plant metabolism. These phytochemicals are
The mosquito repellency response to 2.5 mL of C. odorata used by plants for various biological processes, such as
essential oil of UFY microcapsule-treated cotton fabric for defense against a multitude of pathogens, parasites,
was observed at a significantly lower rate from 64 ± 5.48% and herbivores. They are also used for attracting insects
to 82 ± 8.37% within 60 min of exposure period when which are good pollinators to increase their productive
compared with the 10 mL of C. odorata essential oil of potential. Furthermore, these phytochemicals can also be
UFY microcapsule-treated cotton fabric (p < 0.05). The used as repellents for some insects like mosquitoes, which
peak mean % repellency was observed at a 50–60-min are known to cause various human diseases. Hence, these
exposure period. There was a decrease in the mean % phytochemicals are being used as natural herbal extracts
repellency noted from 82 ± 8.37% to 80 ± 7.07% at 20–30 as repellents for mosquitoes to prevent mosquito-borne
min exposure period and continued to increase afterward illnesses such as dengue (Park et al. 2005; Dudareva et
(see Figure 3). al. 2013; Muhlemann et al. 2014).
The untreated cotton fabric showed % repellency rates The use of natural herbal extracts is gaining wide
ranging from 22 ± 4.47% to 22 ± 8.36 within 60 min of the popularity in mosquito repellency in the Philippines and
exposure period. The % repellency of the untreated cotton worldwide due to its safety profile and efficacy. These
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Vol. 149 No. 4, December 2020 Property of “Ylang-Ylang”
Figure 3. Rate of mosquito % repellency of urea-formaldehyde microcapsule treated fabric with ylang-ylang
essential oil in concentrations at 10, 2.5, and 0 mL (untreated).
natural herbal extracts are components of a wide variety of repellent activities. Linalool is a monoterpene alcohol
commercially available mosquito repellents in our country, found in nature with a nice to pleasant scent. Syed and Leal
which are applied directly to the exposed skin to prevent (2008) discovered in their study that the odor receptors
mosquito bites. These commercial mosquito repellents that respond to DEET are the same odor receptors being
in the form of lotions and sprays contain essential oils activated by linalool through their investigation of the
from lavender, peppermint, basil, eucalyptus, chamomile, electrophysiological effect of repellents on mosquito
lemongrass, geranium, citronella, cedar, and pine trees. olfactory receptors. Interestingly, a study by Park and
However, the use of ylang-ylang essential oil in these colleagues (2005) reported that linalool can enhance up
products is not in demand due to its cost, despite its proven to 92% the protection from mosquito bites and for around
efficacy in mosquito repellency (Pastor 2010; Soonwera 1 h based on the human forearm bioassay they performed
and Phasomkusolsil 2015). using Culex pipiens pallens. Additionally, the mechanism
of action of linalool in acetylcholinesterase inhibition
Several studies have shown the efficacy of ylang-ylang is well-understood. Acetylcholine is a neurotransmitter
(C. odorata) essential oil as a mosquito repellent (Saedi responsible for normal nerve conduction. It is degraded
and Crawford 2006; Phasomkusolsil and Soonwera 2011; by the enzyme acetylcholinesterase. Inhibition of
Cheng et al. 2012). The essential oil of C. odorata contains this enzyme disrupts normal neurotransmission by
several compounds. The principal compounds include inactivating acetylcholine. It has been shown that linalool
linalool, linalool acetate, α-pinene, eugenol, α-terpineol is a reversible inhibitor of acetylcholinesterase and
acetate, isobornyl acetate, α-terpineol, camphor, has the ability to disrupt neurotransmission in insects.
β-caryophyllene, γ-muurolene, humulene, geranyl acetate, Furthermore, linalool occupies at least the hydrophobic
bergamotene, benzyl benzoate, benzyl acetate, methyl site of the enzyme's active center, eliciting its mechanism
benzoate, ρ-methylanisole, and germacrene D – which (Ryan and Byrne 1988).
can be found in the leaves and the flowers of ylang-ylang
(Caballero-Gallardo et al. 2011; Cheng et al. 2012). The use of mosquito repellents applied to the skin for
Furthermore, it also contains 4% farnesol, which is a protection from mosquito bites using natural herbal
sensitizing skin irritant. Thus, only 2% is the maximum extracts or using commercial products containing
safe concentration for C. odorata essential oil to be used as these natural herbal extracts is just one of the many
botanical repellents (Maia and Moore 2011). However, the environmental controls being explored in curbing the
group of compounds responsible for its known mosquito increasing prevalence of mosquito-borne diseases. The
repellency including larvicidal property is due to its advancement in textile and clothing manufacturing
linalool family of compounds. industries have incorporated the use of mosquito repellents
applied directly to textiles and clothing fabrics. These
Linalool is a competitive and reversible inhibitor of mosquito repellents are synthetic chemicals or naturally
acetylcholinesterase, and is one of the potential bioactive occurring compounds from herbal sources. Mosquito-
compounds found in the essential oil of C. odorata that repellent textiles are now being used in bags, mosquito
accounts for the insecticidal, larvicidal, and mosquito
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Vol. 149 No. 4, December 2020 Property of “Ylang-Ylang”
nets, tents, and military apparel. skin irritation. Formaldehyde can cause skin irritation
and ill health effects in the form of fumes and when the
Mosquito-repellent fabrics use microencapsulation, which air concentration exceeds 1 ppm. Microencapsulation
is a technique used in clothing manufacture. This new provides slow disintegration of urea-formaldehyde
and innovative technology uses various microcapsules polymers and circumvents the fume formation of
that act as microscopic containers of mosquito repellent formaldehyde. However, the use of urea-formaldehyde
compounds and are applied directly to the fabric. These microencapsulation of ylang-ylang (C. odorata) essential
microcapsules release their core contents, which could be oil in cotton fabrics for clothing manufacture should be
of synthetic origin or from natural herbal extracts under subjected to rigorous application studies.
controlled conditions to suit a specific purpose (Malik et
al. 2020). Recent studies already show this technique to be
effective in the insect repellency activities of natural herb
extracts such as cedarwood, eucalyptus, and citronella oil
CONCLUSION
(Sharma and Goel 2018; Türkoğlu et al. 2020).
The study showed that the mosquito repellent ylang-ylang
Microencapsulation also circumvents the disadvantage (C. odorata) can be applied to cotton fabrics using the urea
of skin irritation, some health side effects, and unwanted and formaldehyde microencapsulation method. Based on
fabric coloring and staining using natural herbal extracts. the SEM characteristics of the treated fabrics, irregularly
Additionally, it also improves the potency by preventing shaped microcapsules ranging from 50–100 μm in size
evaporation and controlling volatility, which helps in adhered to the treated cotton fabrics. Using the WHO
stabilizing the compounds by protecting them from cone test, the study revealed that the urea-formaldehyde
oxidation that may be caused by moisture, heat, and microcapsule with C. odorata essential oil-treated cotton
contact with other substances (Misni et al. 2017). This fabrics showed significantly higher mosquito repellency
study has proven the utility of microencapsulation using against adult female Aedes aegypti mosquitoes than
urea and formaldehyde as the shelling compound, and the untreated fabric. The treated fabrics with a higher
the C. odorata essential oil as the core compound with concentration of C. odorata essential oil (10 mL) showed
mosquito repellent property that can be applied to clothing significantly greater mosquito repellency than the treated
fabrics. fabric with a lower concentration of C. odorata essential
This repellent activity, however, may diminish after the oil (2.5 mL). This study suggests the potential of urea-
cloth is washed multiple times. This study was limited formaldehyde microencapsulation as an effective method
only to the mosquito repellency in unwashed cotton fabric, in preserving the insect repellent property of C. odorata
and binding agents were not employed for the industrial essential oil and can be a potential mosquito repellent to
finish. However, a simple coacervation method is shown be used in clothing fabric.
to decrease aroma after up to ten washing cycles. The use
of natural herbal extracts as mosquito repellents directly
applied to the fabric has less applicability due to its
limited durability. However, the use of microencapsulation STATEMENT ON CONFLICT OF
can improve the durability of fabrics despite multiple INTEREST
washing. The possible harmful effects of either the The authors declare no conflict of interest in this study.
shelling polymers for microencapsulation or the mosquito
repellents in the core of the microcapsules can also be
lessened when applied to clothing fabrics than direct
application of these on the skin (Raja et al. 2015).
REFERENCES
Although the harmful effects of urea-formaldehyde ABOU-DONIA MB, GOLDSTEIN LB, DECHOVS-
polymers used in microencapsulation were not determined KAIA A, BULLMAN S, JONES KH, HERRICK EA,
in this study, possible harmful effects of shelling polymers ABDEL-RAHMAN AA, KHAN WA. 2001. Effects
like urea and formaldehyde in microcapsules applied to of daily dermal application of DEET and permethrin,
clothing fibers can lessen the potentially harmful effects alone and in combination, on sensorimotor perfor-
such as allergies and skin irritation by the process of mance, blood-brain barrier, and blood-testis barrier in
microencapsulation (Geethadevi and Maheshwari 2015). rats. Journal of Toxicology and Environmental Health
The concentration of urea-formaldehyde polymers Part A 62(7): 523–541.
in microencapsulating the essential oil of C. odorata
adsorbed onto the cotton fabric used in this study, CABALLERO-GALLARDO K, OLIVERO-VERBEL J,
although not determined, may not be enough to cause STASHENKO EE. 2011. Repellent activity of essential
1035
Philippine Journal of Science Reyes et al.: Mosquito Repellent
Vol. 149 No. 4, December 2020 Property of “Ylang-Ylang”
oils and some of their individual constituents against MALIK T, GADODIYA K, KUMAR A. 2020. Mos-
Tribolium castaneum Herbst. Journal of Agriculture quito repellent textiles: an overview. Retrieved on
and Food Chemistry 59: 1690–1696. 20 Feb 2020 from https://technicaltextile.net/articles/
mosquito-repellent-textiles-2574
CHENG J, YANG K, ZHAO NN., WANG XG, WANG
SY, LIU ZL. 2012. Composition and insecticidal ac- MIRO SPECOS MM, GARCIA JJ, GUTIERREZ AC,
tivity of the essential oil of Cananga odorata leaves HERMIDA LG. 2017. Application of microencap-
against Sitophilus zeamais motschulsky (Coleoptera: sulated biopesticides to improve repellent finishing
Curculionidae). Journal of Medicinal Plants Research of cotton fabrics. The Journal of The Textile Institute
6(18): 3482–3482. 108(8): 1454–1460.
DAS NG, BURUAH I, TALUKDAR PK, DAS SC. 2003. MISNI N, NOR ZM, AHMAD R. 2017. Repellent effect
Evaluation of botanicals as repellents against mosqui- of microencapsulated essential oil in lotion formula-
toes. Journal of Vector Borne Diseases 40(1–2): 49–53. tion against mosquito bites. Journal of Vector Borne
Disease 54: 44–53.
[DOH] Department of Health. 2017. DOH urges public
to protect themselves from mosquito bites to prevent MUHLEMANN JK, KLEMPIEN A, DUDAREVA N.
Japanese Encephalitis. Retrieved on 18 Nov 2019 from 2014. Floral volatiles: from biosynthesis to function.
https://www.doh.gov.ph/node/11110&hl=en&gl=ph& Plant, Cell, and Environment 37: 1936–1949.
strip=1&vwsrc=0
PARK BS, CHOI WS, KIM JH, KIM KH, LEE SE. 2005.
[DOST] Department of Science and Technology. 2016. Monoterpenes from thyme (Thymus vulgaris) potential
The mosquito menace in the Philippines. Retrieved on mosquito repellents. Journal of American Mosquito
18 Nov 2019 from http://www.pchrd.dost.gov.ph/in- Control Association 21: 80–83.
dex.php/news/library- health-news/5689-the-mosquito
PASTOR P. 2010. A parent’s anti-mosquito arsenal. Re-
menace-in-the-philippines
trieved on 08 Feb 2020 from https://lifestyle.inquirer.
DUBEY R, SHAMI TC, BHASKER RAO KU. 2009. net/byline/pam-pastor/
Microencapsulation Technology and Applications.
PHASOMKUSOLSIL S, SOONWERA M. 2011. Com-
Defense Science Journal 59: 82–95.
parative mosquito repellency of essential oils against
GEETHADEVI R, MAHESHWARI V. 2015. Long last- Aedes aegypti (Linn.), Anopheles dirus (Peyton and
ing UV protection and mosquito repellent finish on Harrison) and Culex quinquefasciatus (Say). Asian Pa-
bamboo/ tencel blended fabric with microencapsulated cific Journal of Tropical Biomedicine 1(1): S113–S118.
essential oils, Indian Journal of Fibre and Textile Re-
RAJA ASM, KAWLEKER S, SAKENA S, ARPUTH-
search 40: 175–179.
ARAJA A, PATIL PG. 2015. Mosquito protective
DUDAREVA N, KLEMPIEN A, MUHLEMANN JK, textiles-a review. International Journal of Mosquito
KAPLAN I. 2013. Biosynthesis, function, and meta- Research 2(4): 49–53.
bolic engineering of plant volatile organic compounds.
RYAN FM, BYRNE O. 1988. Plant-insect coevolution and
New Phytologist 198: 16–32.
inhibition of acetylcholinesterase. Journal of Chemical
JAJPURA L, SAINI M, ABHILASHA R, CHHICHHO- Ecology 14: 1965–1975.
LIA K. 2015. A review on mosquito repellent finish
SHARMA R, GOEL A. 2018. Development of insect
for textiles using herbal extracts. International Journal
repellent finish by a simple coacervation microencap-
of Engineering Science & Management Research 2(8):
sulation technique. International Journal of Clothing
16–24.
Science and Technology 30: 152–158.
LI S, LEWIS JE, STEWART NM, QIAN L, BOYTER H.
SAEDI N, CRAWFORD GH. 2006. Botanical briefs:
2008. Effect of finishing methods on washing durabil-
ylang-ylang oil – extracts from the tree Cananga
ity of microencapsulated aroma finishing. The Journal
odorata. Cutis 77(3): 149–150.
of the Textile Institute 99(2): 177–183.
SOONWERA M, PHASOMKUSOLSIL S. 2015. Efficacy
LAZKO J, POPINEAU Y, LEGRAND J. 2004. Soy gly-
of Thai herbal essential oil as green repellent against
cinin microcapsules by simple coacervation method.
mosquito vectors. Acta Tropica 142: 127–130.
Colloids and Surfaces Biointerfaces 37: 1–8.
SPECOS MM, GARCÍA JJ, TORNESELLO J, MARINO
MAIA MF, MOORE SJ. 2011. Plant-based insect repel-
P, VECCHIA MD, TESORIERO MVD, HERMIDA
lents: a review of their efficacy, development and
LG. 2010. Microencapsulated citronella oil for mos-
testing. Malaria Journal 10(1): S11.
quito repellent finishing of cotton textiles. Transactions
1036
Philippine Journal of Science Reyes et al.: Mosquito Repellent
Vol. 149 No. 4, December 2020 Property of “Ylang-Ylang”
of the Royal Society of Tropical Medicine and Hygiene [WHO] World Health Organization. 2013a. Guidelines
104(10): 653–658. for testing mosquito adulticides for indoor residual
spraying and treatment of mosquito nets. Retrieved
SYED Z, LEAL WS. 2008. Mosquitoes smell and avoid
on 18 Nov 2019 from https://apps.who.int/iris/han-
the insect repellent DEET. Proceedings of the National
dle/10665/69296
Academy of Science 105(36): 13598-13603.
[WHO] World Health Organization. 2013b. Guidelines
TAN LTH, LEE LH, YIN WF, CHAN CK, KADIR HA,
for Efficacy Testing of Spatial Repellents. Retrieved on
CHAN KG, GOH BH. 2015. Traditional uses, phy-
18 Nov 2019 from https://apps.who.int/iris/bitstream/
tochemistry, and bioactivities of Cananga odorata
handle/10665/78142/9789241505024_eng.pdf
(ylang-ylang). Evidence-based Complementary and
Alternative Medicine 2015: 1–31. [WHO] World Health Organization. 2017. Vector-borne
diseases. Retrieved on 18 Nov 2019 from http://www.
TÜRKOĞLU GC, SARIIŞIK AM, ERKAN G,
who.int/newsroom/fact-sheets/detail/vector-borne-
YIKILMAZ MS, KONTART O. 2020. Micro- and
diseases
nano-encapsulation of limonene and permethrin for
mosquito repellent finishing of cotton textiles. Iranian
Polymer Journal. p. 1–9.
1037