Sugar Apple (Annona squamosa Linn.

) Leaves
as Mosquito Repellent Coil

April Je-ann P. Custodio

Abstract: This study aims to maximize the use of sugar apple (Annona squamosa Linn.) leaves by formulating
a mosquito repellant coil. The gathered sugar apple leaves were washed thoroughly with distilled water.
Then, another set of leaves was heated with 40°C to be oven dried. The 3.13 g of fresh sugar apple leaves,
6.25 g of dried sugar apple leaves, 12.5 ml of ethyl alcohol, 8 ml of distilled water, 1.26 g of grated candle, and
1.68g of cornstarch was mixed together. After mixing, the Japanese paper was cut into long strands and
formed into coil-like shape as foundation of the blended materials. And then the formed coil was put into
oven with temperature of 85°C. With the use of phytochemical screening, the presence of alkaloids which
have nitrogen atom that can be used in insect-repellent, flavonoids which have been known to protect plants
against insects and has potentials as mosquito repellent, and unsaturated sterols which have been reported
to affect insects in different ways including antifeedant, cytotoxic and SCP inhibitor activities, was revealed.
The experimentation between groups was done in three trials of three enclosure of mosquitoes with the
same time intervals. The sugar apple leaves as mosquito repellent coil was tested to have 72% effectiveness
in repelling mosquitoes. Using Analysis of Variance, the result showed that there is no significant difference
between groups, accepting the null hypothesis. This indicates that sugar apple (Annona squamosa Linn.)
leaves can be used as mosquito repellent coil.

Keywords: Sugar apple, mosquito, repellent, coil

2018, according to the Dengue Disease

INTRODUCTION
Mosquitoes are one of the deadliest
animals in the world. The diseases that
mosquitoes carry and transmit to people they
bite, on the other hand, kill 725,000 (Gates,
2014). Mosquitoes are well known transmitters
which transmits various vector-borne diseases
like malaria 15, Dengue, Filariasis, Japanese
encephalitis, Yellow fever, Chickunguniya etc in
humans (Bavani & Srimathi, 2015). The
worldwide incidence of dengue has risen 30-
fold in the past 30 years, and more countries
are reporting their first outbreaks of the
disease. Zika, dengue, chikungunya, and yellow
fever are all transmitted to humans by the
Aedes aegypti mosquito (World Health
Organization, 2019).
Philippines is a country that is prone to
mosquito-borne diseases, like dengue. Dengue
cases in the country are uncontrolled every
year. In the number of persons diagnosed with
dengue, many have died. In the first quarter of
Surveillance Report, the Department of Health
had tallied a number of 20,108 numbers of
dengue cases dropped out in the country from
January 1- March 10. The dengue cases in
Pangasinan increased by 136 percent. A total of
106 386 dengue cases have been reported
nationwide from 1 January to 8 September
2018 (World Health Organization, 2018).
There are many researches that
resulted to effective mosquito repellent. The
most commonly use mosquito repellent is coil
(Gul, Ibrahim, Wasif, Zafar, & Syed, 2013).
1.1 Sugar Apple (Annona squamosa Linn.)
An insecticide plants are now developed
because of their potential to have an
insecticidal effects, most of these plants are
from the family of Meliaceae, Rutaceae,
Asteraceae, Labiatae, Canellaceae, and
Annonaceae (Miller & Koehler, 2008).
The ethanolic extract both from leaves
and seeds of the family of Annonaceae have
many researched that it has efficacy against

1
some insects, especially of vector diseases
mosquito (Chorge, 2012). So the researchers of
this study will use sugar apple leaves as the
major source to have mosquito repellent.
Sugar apple or Annona Squamosa is
belonging to Annonaceae Family. It was also
introduced and became popular in the
Philippines which is called “atis”. This tree has
several medicinal uses, especially its leaves.
According to different studies, the leaves are
having insecticidal and mosquitocidal properties
which is capable to kill insects and pests. One of
these properties is called borneol. According to
Duke (2008), borneol has the possibility of
effectiveness as insecticides or repellent. It is a
essential oil with bicyclic organic compound
that belong to essential oil and has a natural
insect repellent.
In addition, the atis leaves also
possessed anti-pediculosis or lice. It was also
proven in killing cockcroaches (periplaneta
americana). Annona squamosa leaves revealed
to possess different mosquito larvicidal
effectiveness against C. quinquefasciatus. The
leaves are also having acrid principle which
helps to repel mosquitoes because of its
hesitating aroma. Extract from Annonaceae
Family have been tested for control of
Lepidoptera, Hymenoptera, Coleoptera, and
Diptera, especially against Spodoptera
frugiperda, Plutella xylostella, Aedes aegypti
and stored grain insects.
According to Ghosh, Chowdhury, and
Chandra (2012), exploring floral biodiversity and
entering the field of using safer insecticides of
botanical origin has the most effective
alternative approach in a simple and
sustainable method of mosquito control.
Because of this the researchers of this study
aims to have eco-friendly, biodegrable, and
affordable mosquito repellent with the use of
sugar apple or atis leaves as the main variable
of this study.
The researchers moved to conduct this
study to contribute help in decreasing the
number of mosquito-borne diseases by
protecting our families away from mosquitoes.
The researchers were inspired to maximize the
2
use of sugar apple leaves as mosquito repellent
coil to provide people with a cheaper yet
effective product available in the market.
1.2 Why mosquito (Culex quinquefasciatus)?
Culex quinquefasciatus is known as
southern house mosquitoes and is common in
Asia (Connelly, 2013). The larvae feed on biotic
material in the water and require between five
to eight days to complete their development at
300C. Male mosquitoes of this type feed on
blood as well (Buss, Leal, & Nikbakhtzadeh,
2016)
1.3 Conceptual Framework
The plant Annona squamosa Linn. has
been the subject of study in other countries
that is tropical in nature. It is a very popular
plant in Asian countries such as Philippines and
India, to name a few. Since this plant mostly can
be found on tropical countries, a study
conducted on India by Tri Wulandari
Kesetyaningsih from Department of
Parasitology Faculty Medicine and Health
Science stated that Annona squamosa leaves
extract can kill cockroaches. Kesetyaningsih’s
research had used experimental method
consisting seven groups treatment based on the
concentration of the leaf extract (100%, 75%,
25%, 10%, 50%, 5% and 0.1%). As a result,
Annona squamosa leaves extract with 100%,
75%, 50%, and 25% concentration proved to be
effective to kill cockroaches but leaves with
extract 0.1%, 5% and 10% proved to ineffective
to kill the cockroaches (Kesetyaningsih, 2012).
Another study conducted by Rajya
Lakshmi, et. al (2016) also proved the
effectiveness os sugar apple leaves. They
formulated natural mosquito paper coil with
Annona squamosa L. extracts and evaluated its
mosquitocidal activity. In their research, they
conducted that the significant activity
demonstrated by extract of Annona squamosa
suggests that the plants has high killing effects
on mosquitoes (Lakshmi, Mounika, & Babu,
2016).
Doctors from the Philippines also
formulated a mosquito repellent. Dr. Lleana
Cruz and Dr. Lilian de Las Llagas, both from UP
Manila, developed a Atis Lotion Mosquito
Repellent, from atis leaves. It helps to protect
skin from mosquito bites and mosquito-borne
diseases like dengue, malaria, filariasis. The
lotions were now already on pharmacy shelves.
(Aclan, 2012)
A study has been conducted towards
Pediculosis infestation. The study is entitled
“The Efficacy of Atis Leaves Extract as an
Alternative Treatment for Pediculosis” which
aims to lessen lice, which is recognized as one
of the problems of the citizens (Ballesteros,
Basco, & Briones, 2014). Recently, a
phytochemical analysis in the laboratory of the
Department of Science and Technology was
conducted and it was proven that the leaves
contain anthraquinones, which is derivative of
anthracene that can be used as insecticide. This
test proves that the Atis leaves extract contains
an insecticidal property.
It is in this context that this study is
conducted to find out the effectiveness of sugar
apple (Annona squamosa Linn.) leaves as
mosquito-repellent coil. In this paradigm of the
study is shown in figure 1 where the input deals
with the sugar apple leaves while the process
through plant identification, phytochemical
screening and experimentation on the
effectiveness of sugar apple as mosquito
repellent and the output as sugar apple leaves
as mosquito repellent coil.
3
1.4 Objectives of the Study
The objective of this study was to
determine the level of effectiveness of sugar
apple leaves as mosquito repellent.
Specifically, this study was intended for
the following:
1. To determine the active constituents of sugar
apple (Annona squamosa Linn.) leaves as
mosquito repellent coil.
2. To determine the level of effectiveness of the
sugar apple leaves as mosquito repellent coil.
3. To determine the significant difference
between sugar apple leaves as mosquito
repellent coil and the two comparable groups.
1.5 Null Hypothesis
There is no significant difference
between the effectiveness of sugar apple leaves
as mosquito repellent coil and the two
comparable groups.
1.6 Scope and Delimitation
This study focused on the effectiveness
of sugar apple leaves as mosquito-repellent coil.
The researchers have conducted a laboratory
analysis to determine the effectiveness of sugar
apple leaves. The sugar apple leaves was taken
from Baranggay Pagal, San Carlos City,
Pangasinan. The plant identification of the sugar
apple (Annona squamosa Linn.) was conducted
by the Bureau of Plant Industry in Guisad,
Baguio City. The phytochemical screening was
conducted at the College of Pharmacy in Virgen
Milagrosa University Foundation. The mosquito
(Culex quinquefasciatus) larvae were collected
from the stagnant water near the pond of
Virgen Milagrosa University Foundation. The
formation of coil was done in VMUF Senior High
School Biology Laboratory. The experimentation
was conducted at College of Pharmacy in Virgen
Milagrosa University Foundation. This study was
conducted in the year 2018.
1.7 Significance of the Study
The outcome of this study will give
insights to those who will benefit and find its
true meaning to the following entities:
Baranggay. This study will benefit the barangay
to avoid dengue cases and to have alternative
remedy in fighting mosquitoes in their place.
School. This study will also contribute a help to
different schools. As mosquitoes also present in
school the produce product of this study can be
use as alternative coil against mosquitoes.
Householders. This study will help families in
preventing mosquitoes in their houses and to
protect their loveones in mosquito bites and
diseases.
Future Researchers. This study will give big
concepts and ideas to the next batch of Grade
12 students and aspiring researchers in their
future research study, it can also be expounded
by the next researchers.
METHODOLOGY
2.1 Research Design
This study is experimental in nature.
Experimentation was conducted to test the
hypothesis and formulates the conclusion about
the variables.
2.2Materials and Methods
A. Gathering of Sugar Apple Leaves
The researchers gathered fresh sugar
apple leaves in Barangay Pagal, San Carlos
City, Pangasinan.
B. Plant Identification
The plant identification of sugar apple
(Annona squamosa Linn.) was conducted by
the Bureau of Plant Industry in Guisad,
Baguio City. The authenticity of the plant
was proved by the certificate awarded.
C. Procedure of Phytochemical Screening
Through the hot maceration method
using Soxhlet apparatus, the procedure
started. Freshly collected plant materials
were dried in shade and then coarsely
powdered in a blender. 100 g of the coarse
powder will be extracted successively with
250 ml of various solvents in a Soxhlet
apparatus for 24 h. All the extracts were
4
filtered through filter paper separately and
all the extracts were subjected to
qualitative tests for the identification of
various phytochemical constituents as per
the standard procedures (Brinda et al.,
1981; Anonymous, 1990; and Lala, 1993).
The extracts were concentrated in a rotary
evaporator. The concentrated extracts were
used for further analysis. The chemical tests
for various phytoconstituents present in the
extracts were carried out as test for
alkaloids, anthraquinone, catechin,
coumarin, flavonoid, phenol, and quinone.
D. Mosquito Culture
The mosquito (Culex quinquefasciatus)
larvae were collected from the stagnant
water near the pond of Virgen Milagrosa
University Foundation. The collected larvae
were cultivated for seven days in the large
tub containing fresh water (Ramanibai &
Velayutham, 2014). After seven days, larvae
were developed into pupae and further
grown into adult mosquitoes. These
developed mosquitoes were introduced
into three different enclosure.
E. Formation of Coil
A set of gathered fresh sugar apple
leaves were washed thoroughly with
distilled water. Then, another set of leaves
was heated with 40°C to be oven dried. The
3.13 g of fresh sugar apple leaves, 6.25 g of
dried sugar apple leaves, 12.5 ml of ethyl
alcohol, 8 ml of distilled water, 1.26 g of
paraffin wax, and 1.68g of cornstarch was
mixed together. After mixing, the Japanese
paper was cut into long strands and formed
into coil-like shape as foundation of the
blended materials. And then the formed
coil was put into oven with temperature of
85°C.
F. Experimentation
 To determine the effectiveness of 3.2 Level of Effectiveness of Sugar Apple
sugar apple leaves as mosquito repellent (Annona squamosa Linn.) Leaves and the Two
coil, the experimental group was tested to Comparable Groups on Mosquitoes
the cultivated mosquitoes (Culex
quinquefasciatus) as well as for the control
group which was conducted at Virgen
Milagrosa University Foundation College
of Pharmacy.
There were three sets of enclosure of
mosquito net containing 12 living
mosquitoes each, cultivated for seven days.
The sugar apple leaves as mosquito
repellent coil as the experimental group
was put inside the first enclosure and tested
for 3 trials with increasing interval of time.
Then the commercially prepared mosquito
repellent coil as the control group was put
inside the second enclosure and tested for 3
trials with increasing interval of time. And 3.3 Significant Difference between the Sugar
the negative control was observed with the Apple (Annona squamosa Linn.) Leaves and the
same incresing interval of time. The results Two Comparable Groups on Mosquitoes
were recorded by the researchers with the
help of the laboratory technician.

2.3 Statistical Treatment

The main tool used in this study was
one way analysis of variance. This was
computed by using the IBM SPSS Statistics 23
and Microsoft Excel 2016.

RESULTS

3.1 Active Constituents of Sugar Apple (Annona

squamosa Linn.)
Table 1 Active Constituents of Sugar Apple (Annona squamosa Linn.) Leaves
Active Constituents Uses
Alkaloids Have nitrogen atom that can be used in insect-repellent.
Flavonoids Have been known to protect plants against insects and has potentials as
mosquito repellent.
Tannins Reduce growth and fecundity of some insect species.
Unsaturated Sterols Have been reported to affect insects in different ways including antifeedant,
cytotoxic and SCP inhibitor activities.

5
DISCUSSIONS
The active constituents present in sugar
apple leaves that contribute the most are
alkaloids (Ulubelen, et al., 2001), flavonoids
(War, et al., 2012), and unsaturated sterols for
the study because they have insect-repellent
property. According to the CAM Research
Journal, a phytochemical analysis in the
laboratory of Department of Science and
Technology was conducted and it was proven
that the leaves contain anthraquinones, which
is a derivative of anthracene that can be used as
an insecticide. This test proves that the Atis
leaves extract contains an insecticidal property
(Department of Science and Technology, 2010).
As can be seen, the experimental group
has obtained 72% effectiveness. With this, the
researchers present that the experimental and
positive control are both safe and effective. But,
Amy Kingston (2018) said that commercial
repellents may contain chemicals, fragrances or
natural substances that can irritate sensitive
skin or have an unpleasant odor. You can whip
up a homemade insect repellent that uses
ingredients family members are not allergic or
sensitive to. If you prefer, you can use only
organic ingredients.
Based on the statistical treatment using
one way analysis of variance, there is no
significant difference of sugar apple (Annona
squamosa Linn.) leaves and the two comparable
groups on mosquitoes since the t-computed
value is lesser than the t-tabular value. Though,
there is no significant difference between the
experimental and control group, natural insect
repellents have been deemed to be minimum
risk. There are some research studies that
suggest insect repellents with synthetic
ingredients may pose health risks, particularly in
regards to effecting the central nervous system
according to Deter Outdoor Skin Protector. The
prolong use of synthetic mosquito repellents
shows many side effects like neurotoxic
hazards, immunotoxic hazards, skin allergy,
seizures, eye irritation, insomnia, impaired
congnitive function, cough, sneezing, headache,
asthma, bronchial irritation, itching, ear, nose
and throat pain, dermatitis, reproductive
6
dysfunction, development impairment and
cancer, death, (Sharma, 2018).
CONCLUSIONS
This study shows that sugar apple
(Annona squamosa Linn.) leaves can be used as
mosquito repellent coil due to the presence of
phytochemicals needed to perform
mosquitocidal activity. This is in line with the
statement from the study of Lakshmi et. al
(2016) where significant activity demonstrated
by extract of Annona squamosa suggests that
the plant has high killing effects on mosquitoes.
RECOMMENDATIONS
Because the study used only an
enclosure of mosquito net and small amount of
samples to test the level of effectiveness of the
sugar apple leaves as mosquito repellent coil, it
is recommended to be tested in a wider setting
with larger number of samples to determine the
circumference of effectiveness. Also, in order to
determine how long does the sugar apple
(Annona squamosa Linn.) leaves as mosquito
repellent coil last, the standard size should be
followed and duration when burning the whole
coil shall be identified. It is recommended for
the future researchers to use sugar apple
(Annona squamosa Linn.) leaves as mosquito
repellent spray while other parts of the tree are
shall be tested whether it has a mosquitocidal
property. Further researches are also
recommended to conduct toxicology test and
determine the health as well as environmental
impacts of sugar apple (Annona squamosa
Linn.) leaves as mosquito repellent coil.
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