Exploring the Potential of Pineapple (Ananas Comosus) Rind as an Alternative

Bio-Based Mosquito Repellent

Boleche, John Michael C. Ortil, Maria Sheirah E.

Compendio, Jake M. Patrona, Jasmine C.
Dagami, Jianne Dk C. Sotto, Carl Velmer Y.
Enano, Riezel Mae V. Suyom, Lhyme C.
Nuñez, Emmanuel
Block 19 - STEM

A Research Project
Submitted in Fulfillment
of the Senior High School Program
Academic Track – STEM

ACLC College of Tacloban

Senior High School Department
Tacloban City, Leyte

2023
 LIST OF FIGURES AND TABLES

Figure No. Title Page

1 Conceptual Framework 3
(IV-DV MODEL)

2 The process of 19
mosquito-repellent
production

Table No. Title Page

1 Materials 16

2 Equipments 16

3 Variables 18

4 Treatments 18

5 Constant Variables 18

6 Determining the most 23

effective concentration in
three replicates

7 Most effective 23
concentration and its
frequency

i
 CHAPTER I

INTRODUCTION

The introductory chapter presents a comprehensive framework that

considers broad and detailed aspects related to the research problem. The

researcher begins by evaluating existing knowledge in relevant fields,

recognizing limitations, and identifying areas that require further exploration.

Background of the Study

Mosquito-borne illnesses, such as jungle fever, dengue fever, and Zika

infection, are significant worldwide health issues, with an estimated 216 million

cases and 435,000 deaths every year (WHO, 2019). Ordinary mosquito

repellents, such as DEET and picaridin, can be applicable but can too have

unfavorable side impacts, such as skin irritation and neurotoxicity according to

Mehlhorn (2016).

According to Eisawy et al. (2016), there has been increasing interest in

developing natural and alternative mosquito repellents that are both effective and

safe for use. Studies by Garg et al. (2018) and Khan et al. (2016) have explored

the use of bromelain derived from Pineapple as a mosquito repellent. However,

limited research has focused on the potential of Pineapple (Ananas comosus)

rind extract as a natural mosquito repellent. Therefore, the present study aims to

investigate the possibility of Pineapple (Ananas comosus) rind as an alternative

mosquito repellent.
 According to Prabhu et al. (2014) and Patra and Panda (2016), there has

been increasing attention towards the use of natural product-based mosquito

repellents, such as essential oils and plant extracts. These alternatives are

gaining popularity as they have the potential to offer effective and safe protection

against mosquitoes, while posing minimal risks to the environment and human

health.

Ananas comosus, sometimes known as the pineapple plant, is a tropical

fruit that is commonly farmed over much of the world, notably in tropical and

subtropical areas (Pandey & Shukla, 2015). The enzyme bromelain, which is said

to have a lot of therapeutic qualities, is abundant in the pineapple rind (Khan et

al., 2016). Numerous research studies have shown that pineapple (Ananas

comosus) bromelain extract has the potential to function as a natural insect

repellent. As a result, using pineapple rind extract as a natural mosquito repellent

has been researched (Garg et al., 2018; Khan et al., 2016). The objective of this

study is to explore the potential of pineapple (Ananas comosus) rind extract as

an alternative and effective bio-based mosquito repellent, with the aim of

providing a natural and sustainable solution to mosquito-borne diseases.

Objectives of the Study

The general objective of this study is to determine the feasibility of using

pineapple (Ananas comosus) rind extract as an alternative bio-based

mosquito-repellent. The study will further investigate the following specific

objectives:

2
 1. To determine the efficacy of the mosquito repellent with the varying

concentration of pineapple rind.

2. To evaluate the repellency of the pineapple rind extract through

cage test.

3. To determine the characteristic of the mosquito repellent in terms

of:

3.1. Acidity

3.2. Alkalinity

Statement of the Hypothesis

Ha: The pineapple rind extract is effective in repelling mosquito.

Ho: The pineapple rind extract is not effective in repelling mosquito.

Conceptual Framework

Figure 1. Conceptual Framework (IV-DV MODEL)

The independent variable (IV) in this study is the concentration of

pineapple rind extract. The concentrations will be changed (50%, 75%, 100%) to

decide the most applicable concentration that can repel mosquitoes. The

dependent variable (DV) in this study is the mosquito repellency, which is able be

measured by the number of mosquitoes that are repelled by the pineapple rind

extract and will be compared with the number of mosquitoes that are repelled by

commercially accessible synthetic mosquito repellents. The study will use an

3
experimental design to compare the effectiveness of pineapple rind extract with

commercially available synthetic mosquito repellents. The data collected will be

analyzed to determine the most effective concentration of pineapple rind extract

as a mosquito repellent.

Significance of the Study

This research presents a lot of advantages that can add to the following

groups and individuals:

Consumers of Mosquito-Repellent Products. Consumers of mosquito

repellent products will benefit from this study as it will provide them with an

alternative, bio-based option that is not only effective, but also made from natural

ingredients. This can provide them with peace of mind knowing that they are not

exposing themselves and their loved ones to‌harmful chemicals.

Pineapple Farmers. Agriculturists who grow pineapples can benefit from this

study because it will give them with an extra source of pay by utilizing the

frequently disposed of pineapple skins as a source of crude material for mosquito

repellent production. This may give them a maintainable way to earn a living and

support their families.

Public Health Officials. Public health officials will benefit from this study as it will

provide them with an additional tool in their fight against mosquito-borne

diseases. By identifying an effective bio-based repellent, they can help to reduce

the spread of diseases such as dengue and malaria.

Environmentalists. Environmentalists will benefit from this study because it will

provide them with an alternative to synthetic repellents, which are known to be

4
harmful to the environment. By recognizing a bio-based repellent, it can offer

help in minimizing the sum number of chemicals that are discharged into the

environment.

Future Researchers. Future Researchers will benefit from this study as it will

provide them with new information on the potential use of pineapple rind as a

source of bio-based repellent. This can lead to further research and development

of new products that can help ‌protect people from mosquito-borne diseases.

Scope and Limitations of the Study

The scope of this study will focus on using pineapple (Ananas comosus)

rind extract as an alternative bio-based mosquito repellent. The study will aim to

determine the feasibility of using pineapple rind extract as a mosquito repellent

by conducting a cage test, to measure its effectiveness in repelling mosquitoes.

Additionally, the study will compare the effectiveness of pineapple rind extract

with commercially available synthetic mosquito repellents. One of the

delimitations of this study is the geographical location in which the research will

be conducted. The study will be limited to Tacloban City, which is within the

Philippines. This limitation is because of the availability of pineapple rind extract

may vary depending on the location, and therefore the results of the study may

not be generalizable to other regions.

Another delimitation of the study is the time frame in which the research

will be conducted. The study will be conducted during the second semester of the

school year 2022-2023. This limitation is due to the availability of resources and

funding, as well as the availability of participants and research facilities. The

5
study will also be limited to using ethanol as a solvent in the extraction process of

pineapple rind extract. This delimitation is because of other solvents may have

different effects on the extract and may alter the results of the study.

Definition of Terms

The definition of terms section in this research paper presents concise

explanations of key terms and concepts essential for understanding the study.

These definitions help ensure a common understanding of terminology and

facilitate clear communication throughout the research.

● Alternative bio-based mosquito-repellent - refers to a natural and safe

repellent that is derived from natural sources such as plants, as opposed

to synthetic chemical repellents.

● Feasibility study - refers to examining the practicality and potential

success of a proposed project or idea.

● Mosquito-repellent - refers to a substance or product used to repel

mosquitoes, preventing them from landing on the skin or entering a

specific area.

● Rind - refers to the outer protective covering of a fruit, in this case, the

pineapple rind.

6
 CHAPTER II

REVIEW OF RELATED LITERATURES AND STUDIES

The Review of Related Literature and Studies surveys existing studies and

research relevant to the current study, highlighting key findings and identifying

research gaps. It serves as a knowledge base that informs the objectives and

contextualizes the current research.

Foreign Literature

Mosquito-borne diseases are a significant public health concern

worldwide. Mosquitoes are vectors for a variety of diseases, including dengue,

malaria, West Nile virus, and Zika virus (World Health Organization, 2020).

These diseases cause significant morbidity and mortality, with an estimated 216

million cases of malaria and 25 million cases of dengue fever globally in 2019

(WHO, 2020). Mosquito-borne diseases excessively affect defenseless

populaces, including children, pregnant ladies, and people living in destitution

WHO, 2020). Successful control of mosquitoes and the diseases they transmit is

crucial for decreasing the burden of disease and progressing public health

(WHO, 2020).

Research has shown that conventional control strategies, such as the

usage of pesticides and mosquito nets, have had constrained success in

decreasing the burden of mosquito-borne diseases (Kilama, 2018). As a result,

there is a need for alternative, appropriate control strategies that can effectively

decrease mosquito populaces and the transmission of diseases (Kilama, 2018).

Vector-borne diseases are a long-term health concern in different regions with

scorching weather. Tropical areas such as South America, Africa, and Asia

usually suffer from these diseases, specifically dengue and Malaria (Corbel et al.,

2013). Mosquito species (Anopheles sp.) and (Aedes aegypti) cause dengue and

Malaria. These are prominent and thriving in regions with humid weather

conditions, such as in Asia and Southeast Asian countries, but they are also

becoming inconvenient in different world regions, such as European countries.

Due to these alarming health risks mosquitos bring, chemical-based repellent is

widely adopted worldwide. However, it is not a concealed fact that

chemical-based insecticides and repellents utilize many harmful elements that

can further compromise public health; because of this, many researchers aimed

to create natural-based repellents to avoid or minimize health risks (Ciera et al.,

2017).

Bio-based mosquito repellent can be derived from different biotic materials

such as plants. Various plant parts are being studied to create an economical but

effective repellent that can combat harmful mosquito attacks in different regions,

such as Asia. In the study conducted by (Shukla et al., 2018), they utilized

plant-based repellent from five different leaves of the Citronella plant group,

namely, Lantana (Lantana camara L.), (Calotropis.), Lemon Grass (Cymbopogon

citratus.), Tulsi (Ocimum sanctum.) and Neem (Azadirachta indica). They

extracted the plant oils from these plants and produced a mixture to be tested.

Their study demonstrated a significant repellent effect against mosquitoes due to

the non- volatile components of the Calotropis plant.

8
 Furthermore, they suggest that researchers investigate different

plant-derived oil varieties to discover other potential natural repellents. Another

study conducted last 2016 investigated the use of various plant oil extracts of the

traditional plants used to repel mosquitoes. The study emphasizes the plant oils

of Catnip (Nepeta cataria.), Citronella (Cymbopogon nardus.), Clove (Syzygium

aromaticum.), Hairy basil (Ocimum americanum.), and Vetiver (Vetiveria

zizanioides.) concerning the Arm-in-cage test as their experimental test. They

determined the plant components that contribute to its mosquito-repellent effect;

this includes Z, Enepetalactone, Citral, Eugenol, Methyl cinnamate, and

terpinen-4-ol, to name a few.

Fruits are also considered an effective biotic material that can combat the

health risks mosquitoes bring. Rafiqi and Nindia (2022) from Indonesia, a part of

the Southeast Asia region, demonstrated this in their experimental study of

pineapple peels. Different concentrations of pineapple peel extracts were applied

in the human hand as mosquito bait in their experiment (10%, 20%, and 30%,

respectively). The experiment outcome demonstrated that a higher concentration

of pineapple rinds could repel more mosquitoes since only a few remained

perched in the experimental hand bait. This conclusion was further supported by

the statistical test (ANOVA) among the various replicates of the experiment.

Hundreds of studies have been reported in recent years describing plant

derivatives that exhibit repellent properties (Zhu et al., 2018). The worldwide

incidence of dengue fever has increased 30-fold in the past 30 years, and more

countries are reporting their first outbreaks of the disease (WHO, 2019). The best

9
strategy to lower the incidence of mosquito-borne diseases is to avoid mosquito

bites using topical repellents and repellent-treated mosquito nets (Govindarajan

et al., 2016). An insect repellent is a substance that acts locally or at a distance

to prevent insects from biting human or animal skin. The repellent provides a

vapor barrier to prevent insects from coming into contact with the skin surface

(Legeay et al., 2018). Further research has been carried out on developing

natural insect-repellent formulations to control Aedis aegypti, which are safer for

humans, the environment, and non-target organisms (Oliveira et al., 2016).

Mosquitoes represent one of the most significant threats to human and

veterinary health worldwide (Norris & Coat, 2017). A repellent generally

discourages arthropods from landing or stinging human skin (Lee, 2018).

Nowadays, the use of mosquito repellent has increased because of dengue,

malaria, and yellow fever, while the main constituent of mosquito repellent is

DEET which is hazardous to human health. The interest in using plants as

mosquito repellents has increased significantly, and one of them is using a

pineapple rind.

Botanical mosquito repellents, which cause little risk to the environment or

human health, may be feasible alternatives to synthetic chemical repellents such

as DEET. Thus, many people prefer to use natural repellents extracted from

plants, such as citronella oil from Cymbopogon nardus, p-menthane-3,8-diol

(PMD) from Eucalyptus maculata citriodora, and fennel oil from Foeniculum

vulgare (Jong et al., 2015). Mosquito-borne diseases can affect people of all

ages, but certain age groups may be more susceptible to infection or more likely

10
to experience severe symptoms. Young children, the elderly, and people with

weakened immune systems are more vulnerable to the severe forms of disease

caused by mosquito-borne viruses, such as malaria, dengue, and yellow fever.

This is because their immune system is not fully developed, or their immune

system is not fully functioning.

Pregnant women and their unborn children are at higher risk of

complications from certain mosquito-borne diseases, such as malaria and Zika.

This is because pregnant women have a weakened immune system, and the

disease can also harm the unborn child. It is important to note that

mosquito-borne diseases can also affect people of any age who have not been

exposed to the disease and have not developed immunity. Dengue diseases are

re-emerging mosquito-derived diseases that are endemic in the Tropics. Dengue

cases have increased in South-East Asia, Africa, the Western Pacific, and the

Americas. The increasingly widespread distribution and the rising incidence of

dengue virus infections are related to the increased distribution of Aedes aegypti,

an increasingly urban population, and increasing air travel. Several Southeast

Asian countries show that the age of the reported dengue cases has increased

from 5-9 years for older children and young adults.

Dengue infection in adolescents and adults has also been recognized as a

potential hazard to international travelers returning from endemic areas,

especially in Southeast Asia (Tantawichien, 2015). Mosquitoes can become

infected by people who are viremic with DENV. This can be someone who has a

symptomatic dengue infection, someone who is yet to have a symptomatic

11
infection (they are pre-symptomatic), and people who show no signs of illness

(they are asymptomatic). (Duong et al., 2015). Dengue is one disease entity with

different clinical presentations, often with unpredictable clinical evolutions and

outcomes. Bleeding manifestations in adult patients, including petechiae and

menorrhagia, were also frequently found; however, massive hematemesis may

occur in adult patients because of peptic ulcer disease and may not be

associated with profound shock as previously reported in children. Although

shock and plasma leakage seems to be more prevalent as age decreases, the

frequency of internal hemorrhage rises as age increases. An increase in liver

enzymes found in children and adults indicated liver involvement during dengue

infections.

Pre-existing liver diseases in adults, such as chronic hepatitis, alcoholic

cirrhosis, and hemoglobinopathies, may aggravate liver impairment in dengue

infection. Fulminant hepatitis is a rare but well-described problem in adult

patients with dengue infection. Currently, no specific therapeutic agent exists for

dengue. The early recognition of dengue infection, bleeding tendency, and signs

of circulatory collapse would reduce mortality rates in adult patients with dengue

infection (Tantawachien, 2015).

The investigation into the toxicity of acids for mosquito larvae has provided

valuable insights into the physiological characteristics of these larvae. It has been

observed that mosquito larvae typically exhibit a neutral or slightly alkaline

reaction, with a pH range of 7.0-7.4. This understanding of their pH preference is

crucial in evaluating the potential effectiveness of acid-based compounds as

12
larvicidal agents. By considering the larvae's natural pH range, researchers can

further explore the use of acids to target and control mosquito populations during

their vulnerable larval stage.

Local Literature

One of the most concerning issues in the Philippines is the predominance

of mosquito-borne diseases such as dengue, jungle fever, and Zika infection.

According to the Office of Wellbeing (DOH), there were over 200,000 detailed

cases of dengue in 2019 alone, resulting in over 800 deaths. The same report

states that the number of detailed cases of malaria within the Philippines has

been steadily expanding over the past few years, with over 6,000 cases detailed

in 2019. The Zika infection has, moreover, been reported within the Philippines,

with over 100 cases reported in 2018.

These diseases not only cause critical health issues for people who

contract them, but moreover result in a critical economic burden for the country.

In expansion to the cost of treating the diseases, there is also a cost related with

lost efficiency due to illness. This highlights the significance of finding effective

ways to control the mosquito population and avoid the spread of these diseases.

In line with this, a few studies have been conducted within the Philippines to

examine the potential of utilizing natural products as mosquito repellents. For

example, a study by Loyola et al. (2018) found that the fundamental oil of lemon

grass (Cymbopogon citratus) had a strong repellent impact against Aedes

aegypti mosquitoes, the primary vector of dengue within the Philippines. Another

study by Asuncion et al. (2019) found that the extract of tubli (Calophyllum

13
inophyllum) clears out had a strong repellent effect against both (Aedes aegypti)

and (Culex quinquefasciatus) mosquitoes.

Foreign Studies

One study, conducted by Xing et al. (2018), examined the effectiveness of

plant-based mosquito repellents. The study utilized lemon eucalyptus oil,

citronella oil, and peppermint oil as natural repellents and compared their viability

to DEET, a commonly utilized synthetic repellent. The results appear to be that all

three plant-based oils give protection against mosquitoes for at least two hours,

with lemon eucalyptus oil providing the longest term of protection at roughly three

hours. In comparison, DEET provided protection for roughly five hours. The study

concluded that plant-based oils may be a appropriate alternative to synthetic

repellents, particularly for those who favor to utilize natural products.

Another study by Patra et al. (2013) which pointed to evaluate the

repellency of essential oils of diverse plants against mosquitoes, the results

appeared that citronella, eucalyptus, lemongrass and peppermint have higher

rate of repellency compared to the control group. The study moreover, found that

eucalyptus oil had the highest repellency effect. Several studies on the use of

ethanol as a solvent for filtering and diluting plant extracts, including pineapple

rind extract, have been conducted. Plant extracts from six medicinal plants,

including pineapple, were tested for larvicidal activity against mosquito larvae by

(Samake et al., 2011). According to the authors, the plant material was extracted

with water and then filtered with filter paper. The filtrate was then concentrated

and diluted with ethanol to the concentration required for testing. The use of

14
ethanol was most likely beneficial in removing any remaining impurities and

adjusting the extract's concentration.

Similarly, Gunasekaran et al. (2015) used pineapple peel extract to

evaluate its repellency against Aedes aegypti mosquitoes. They filtered the

extract using Whatman filter paper and diluted it with ethanol before testing. The

authors suggested that the use of ethanol helped to increase the solubility of the

extract and improve its stability.

In a study conducted by Hossam et al. (2022), the researchers employed

the maceration method to extract Lansat (Lansium domesticum) with the aim of

assessing its potential as a natural larvicide. The maceration method involves

soaking the plant material in a solvent to extract its active compounds. This

approach allows for the extraction of various bioactive constituents present in

Lansat, which can then be evaluated for their larvicidal properties. By exploring

the efficacy of Lansat as a natural larvicide, this study contributes to the ongoing

research on environmentally friendly alternatives for mosquito control.

Local Studies

One study conducted by Reyes and dela Cruz (2020) in the Philippines

evaluated the repellency of different local plants against mosquitoes. The study

found that the leaves of lemongrass (Cymbopogon citratus) and citronella

(Cymbopogon nardus) had the highest repellency against mosquitoes with an

average of 97.3% and 96.5% protection time, respectively. Another local plant

found to have high repellency properties was the orange (Citrus sinensis) with an

average of 93.8% protection time. The study also showed that the repellency of

15
these plants was comparable to commercially available synthetic mosquito

repellents.

In a research conducted by Santos et al. in 2018, the leaves of oregano

were extracted through Direct Heat Infusion using isophryl alcohol as a solvent.

To facilitate the study, the researchers constructed a custom-made glass

mosquito cage measuring 10 x 6.5 x 6.5 inches. They followed a structured

approach, implementing a 30-minute interval during which they carefully

monitored and recorded the number of mosquitoes that either bit the hand or

gathered in a specific alignment. Subsequently, the data was plotted and the

average number of observed mosquitoes was calculated based on three

separate replicates, ensuring a more comprehensive analysis. The researchers

also computed the repellency rate of each replicates resulting to replicate 1 to

have 100% of repellency rate, replicate 2 to have 93% repellency rate, and

replicate 3 to have 93% repellency rate.

16
 CHAPTER III

METHODOLOGY

Materials and Equipment

This segment of the study presents a comprehensive list of the materials

and equipment that will be utilized within the experimental procedure, including

the specific purposes for which each will be utilized.

Table 1. Materials
Materials Uses

Pineapple The primary source of the active compounds used to make the

pineapple rind extract.

Mosquitoes Used as the test subject to determine the effectiveness of the

pineapple rind extract as a mosquito repellent.

Ethanol(70%) Used as a solvent to extract the pineapple rind and to dilute the

solution.

Table 2. Equipments
Equipments Uses

Flask Used to hold the pineapple rind-ethanol mixture during the

extraction process.

Mosquito Cage Used to contain the mosquitoes during the test.

Gloves Worn to protect the hands from contact with the mosquitoes and
 the ethanol during the experiment.

Goggles Worn to protect the eyes from splashes of ethanol during the

experiment.

Laboratory Coat Worn to protect the clothing from contact with the mosquitoes and

the ethanol during the experiment.

Knife Used to cut and prepare the pineapple rinds for extraction.

Coffee Paper Used to filter the pineapple rind-ethanol mixture to obtain the

Filter pineapple rind extract.

Food Processor Used to process the pineapple rinds before extraction.

Filter Funnel Used to hold the coffee paper filter during the filtration process.

Treatment and Variables

The manipulation and measurement of variables are crucial aspects of

experimental design, particularly in entomology. In this study, the independent

variable is the concentration of pineapple rind extract, which is manipulated to

determine its effect on the dependent variable, which is the number of

mosquitoes repelled. In order to ensure the validity and reliability of the results, it

is necessary to control for other extraneous variables, such as the number of

mosquitoes released in the mosquito cage, and the duration of exposure.

Table 3. Variables
Independent Variable Dependent Variable Controlled Variables

17
 ● Concentration ● Number of ● Mosquitos Exposed

of Pineapple Mosquitos in the Cage

Rind Extract Repelled ● Duration of Exposure

Table 4. Treatments
Treatments Concentrated Pineapple Rind

T0 0mL

T1 75mL

T2 100mL

T3 125mL

Table 5. Constant Variables

Ethanol Mosquitoes Released in the Duration of Exposure
Cage

25mL 15 Mosquitoes 2 hours

This study will use four distinct concentrations of pineapple rind extract,

specifically 0mL, 75mL, 100mL, and 125mL with each mixed with 25mL of 70%

ethanol.

18
 Flowchart of the Procedure

Figure 2. The process of mosquito-repellent production

General Procedure

A general procedure acts as a comprehensive guide for carrying out a

specific process and forms the framework of written documentation. This will give

a detailed, step-by-step explanation of the procedures that must be followed in a

structured way, in addition to the materials and equipment required to obtain the

extracted pineapple rind.

1. Preparation of the Pineapple

The extraction of juice from the pineapple rind involved following a series

of procedures prior to the actual extraction process. These procedures include:

● Peeling

● Cutting

● Drying

19
2. Peeling

The selected pineapple underwent a peeling process using a sharp knife

to effectively separate the mesocarp, which was the fleshy edible portion of the

fruit, from the rind, which served as the outer protective layer. This method of

separation was commonly employed in the food industry as it allowed for the

efficient removal of the inedible rind and preservation of the desirable mesocarp.

3. Cutting

The pineapple rinds were meticulously segmented into smaller,

manageable pieces using a knife. This step played a crucial role in the drying

process, as the smaller pieces allowed for a greater surface area to be exposed

to sunlight. Consequently, the drying process became more efficient and

consistent. Moreover, the smaller pieces possessed a higher surface area to

volume ratio, which facilitated faster water loss and ensured a more thorough

drying process.

4. Drying

The cut pineapple rinds were then subjected to the drying process, a vital

step in eliminating any remaining moisture from the rinds. This drying phase was

crucial as it prepared the rinds for the subsequent crushing process. Additionally,

the drying process played an important role in preventing the growth of bacteria

and fungi, which could potentially compromise the quality and safety of the final

product. By effectively removing moisture, the drying process ensured that the

pineapple rinds were suitable for further processing and minimized the risk of

microbial contamination.

20
5. Extraction

During the extraction process of pineapple rind extract, it was important to

prioritize safety precautions to protect against potential hazards. This included

wearing personal protective equipment such as gloves, goggles, and a laboratory

coat. These safety measures helped minimize the risk of skin and eye irritation

caused by contact with ethanol and pineapple rinds. Ethanol was a flammable

solvent that could cause skin irritation, so wearing gloves was essential to

prevent direct contact. Goggles were worn to protect the eyes from solvent

splashes, and a laboratory coat provided protection for clothing. The extraction of

juice from pineapple rind involved following a series of procedures. These

procedures include:

● Crushing and Mixing

● Resting of Solution

● Filtration

6. Crushing and Mixing

The dried pineapple rind was then subjected to a mechanical processing

method, utilizing a conventional gear such as a food processor. This step aimed

to break down the rind into smaller particles and enhance its surface area for

efficient extraction. Subsequently, the processed pineapple rind was mixed with

ethanol at a ratio of 1:10 to facilitate the extraction process through maceration.

The maceration method allowed the ethanol to extract the desired compounds

from the pineapple rind, resulting in the production of the desired extract.

21
7. Resting of Solution

The mixture of processed dried pineapple rind and ethanol was allowed to

rest for a period of 24 hours. This resting period was essential as it facilitated the

separation of impurities and debris from the desired extract. During this time, any

unwanted materials present in the mixture were given the opportunity to settle

and undergo sedimentation.

8. Filtration

After the designated resting period, the mixture of ethanol and processed

dried pineapple rind proceeded to the filtration stage. This involved employing a

filter funnel and coffee paper filter to separate and purify the desired compounds

within the mixture. The filtration process was crucial in achieving the desired

outcome by effectively separating the liquid and solid components. By utilizing

the filter funnel and coffee paper filter, the researchers could efficiently remove

any solid particles or impurities, resulting in a higher quality and purer final

product. This meticulous filtration allowed the researchers to obtain the desired

Pineapple rind extract, ready for further analysis and application in their study.

. Dilution

After filtration, the researchers obtained a 100% concentrated Pineapple

Rind extract. However, to conduct subsequent experiments and evaluate the

extract's effectiveness at different concentrations, it was necessary to obtain

diluted extracts at 75%, 80%, and 83.3% concentrations. The 100% concentrated

extract was diluted with ethanol at specific ratios through a dilution process to

achieve these desired concentrations. This process allowed the researchers to

22
manipulate the concentration of the extract, enabling comprehensive evaluations

of its efficacy at various concentrations.

Data Gathering

This section of the study is dedicated to outlining the procedure for data

collection. The researcher will employ structured observations employing tables

3.1 and 3.2, which are composed of predetermined categories. These tables will

serve as a framework for researchers to organize and document the

observations made during the experiment. Using structured observations and

predefined categories allows for a systematic and consistent approach to data

collection, increasing the results' reliability and validity.

Table 6. Determining the most effective concentration in three replicates

Time Treatment 1 Treatment 2 Treatment 3 Treatment 4
Interval

30 Minutes

60 Minutes

90 Minutes

Table 7. Most effective concentration and its frequency

Time Control Experimental
Interval (Untreated) (Treated)

30 Minutes

60 Minutes

90 Minutes

The experiment will be conducted simultaneously in Tables 6 and 7 to

compare the effectiveness of the Pineapple Rind solution and a control treatment

23
regarding the frequency of mosquito landings on the treated areas. To compare

the relative effectiveness of each, the researchers will first determine the best

concentration of the pineapple rind extract, which will then be compared

concurrently with the control treatment with 3 replicates..

Data Analysis

Descriptive Statistics

The researchers calculated the measures of central tendency, including

the mean, to analyze the gathered data. The mean provided an average value,

offering insights into the overall trend or concentration of the pineapple rind

extract. Additionally, the researchers employed the calculation of standard

deviation to evaluate the spread or dispersion of the data, providing a clear

understanding of the consistency or variability of the extract's effectiveness as a

mosquito repellent. These calculations assisted the researchers in gaining

insights into the data and assessing the potential of pineapple rind extract as a

mosquito repellent.

Measures of Central Tendency.

Measures of Variability.

Inferential Statistics

24
 The researchers plan to compare the most effective concentration of

pineapple rind extract to the control group. The researchers will use the t-test

statistical method to determine whether or not there is a significant difference

between the two groups.

T-Test.

For the hypothesis testing, the researchers will use an ANOVA (Analysis of

Variance) statistical method. It will follow the decision rule where if computed

value is less than the f-critical value at d.f(degrees of freedom) and at specific

level of significance; Accept the null hypothesis; and if the f-value is greater than

f-critical value at d.f and specific level of significance; reject the null hypothesis.

Analysis of Variance (ANOVA).

Parameters

The researchers investigated the potential of pineapple rind extract as an

alternative bio-based mosquito repellent in the study. The primary outcome

measure was the percentage of mosquito bites avoided by the extract. To assess

the effectiveness of the extract, the researchers utilized the following parameters:

25
1. Concentration: The concentration of pineapple rind extract have been

varied to determine the optimal concentration for maximum mosquito

repellency.

2. Repellency rate: The researchers have calculated the repellency rate of

the extract for each concentration.

3. pH level: The pH level of the extract have been varied to determine the

optimal pH level for maximum mosquito repellency.

a. pH level range of 0 to 5 is Acidic

b. pH level of 7 is Neutral

c. pH level range of 8 to 14 is Alkaline

26
 References

Abdallah, H. M., Mohamed, G. A., & Ibrahim, S. R. M. (2022, April 6). PDF.
Basel; MDPI.

Akbari, J., Sheikh, A. R., Hossain, M. J., & Akbar, M. S. (2014). Mosquito
repellent activity of some essential oils and their constituents against
Aedes aegypti L. Asian Pacific Journal of Tropical Biomedicine, 4(8),
633-638.

Añez, G., Volkova, E., Jiang, Z., Heisey, D. A., Chancey, C., Fares, R. C., ...
& Collaborative Study Group. (2017). Collaborative study to establish
World Health Organization international reference reagents for dengue
virus Types 1 to 4 RNA for use in nucleic acid testing. Transfusion, 57,
1977-1987.

Arora, D., Kaur, H., Kaur, M., & Kaur, G. (2014). Mosquito larvicidal, pupicidal
and repellent activity of bromelain from pineapple (Ananas comosus)
against Culex quinquefasciatus. Parasitology research, 113(2), 767-772.

Asuncion, R. V., Corpuz, M. L., & Ocampo, R. C. (2019). Repellent activity of

Calophyllum inophyllum leaf extract against Aedes aegypti and Culex
quinquefasciatus mosquitoes. Journal of Vector Borne Diseases, 56(2),
131-136.

Department of Health. (2019). Epidemiology Bureau.

Duong, V. et al. (2015), Asymptomatic humans transmit dengue virus to

mosquitoes. Proceedings of the National Academy of Sciences of the
USA, 2015. 112(47): p. 14688–14693.

Eisawy, M. A., Al-Ansari, A., & El-Sayed, K. M. (2016). Mosquito repellents: a

review. Journal of advanced research, 7(2), 171-182.

Garg, R., Srivastava, S., & Dixit, V. K. (2018). Mosquito repellents: current
scenario and new horizons. Journal of vector borne diseases, 55(4),
201-210.

Gonzalez, J. & Santos, A. (2019). Repellency of Essential Oils Extracted from

Local Plants Against Mosquitoes. Philippine Journal of Science, 148(2),
117-123.
Gunasekaran, K., Sivakumar, R., Rajeswary, M., & Govindarajan, M. (2015).
Evaluation of plant extracts for mosquito repellency against Aedes aegypti
(L.) (Diptera: Culicidae). Parasitology Research, 114(2), 747-753.

27
Khan, R., Ali, A., & Khan, R. (2016). Potential of bromelain as a natural mosquito
repellent. Journal of Vector Borne Diseases, 53(4), 243-247.

Kilama, W. L. (2018). Mosquito-borne diseases and their control in Africa: A

review. Journal of Vector Borne Diseases, 55(2), 53–67.

World Health Organization. (2020). Mosquito-borne diseases.

Loyola, J. D., Corpuz, M. L., & Ocampo, R. C. (2018). Repellent activity of

Cymbopogon citratus (lemon grass) oil against Aedes aegypti (Diptera:
Culicidae). Journal of Vector Borne Diseases, 55(1), 39-44.

Mehlhorn, H. (2016). Insect repellents and their application. Parasitology

Research, 115(5), 1771-1782.

Pandey, B., & Shukla, S. (2015). Pineapple: A versatile fruit. International Journal
of Pharmacy and Pharmaceutical Sciences, 7(6), 1-7.

Parmar, N., & Kaur, H. (2017). Mosquito repellent potential of essential oils: a
review. Journal of Vector Borne Diseases, 54(4), 261-270.

Patra, R.D., Bhatt, R.P., & Dhiman, R.C. (2013). Repellency activity of different
essential oils against mosquitoes. Asian Pacific Journal of Tropical
Medicine, 6(9), 683-688.

Patra, J. K., & Panda, T. (2016). Mosquito repellent activity of essential oils: a
short review. Journal of vector borne diseases, 53(1), 1-6.

Prabhu, K., Rajeswary, M., & Chandrasekar, A. (2014). Mosquito repellent activity
of essential oils and their major compounds against Culex
quinquefasciatus Say (Diptera: Culicidae). Journal of Vector Borne
Diseases, 51(1), 15-22
Rafiqi, & Yuni Nindia, Y. N. (2022). THE EFFECTIVENESS OF PINEAPPLE
PEEL EXTRACT (Ananas comosus) AS A MOSQUITO REPELLENT FOR
AEDES AEGYPTI. ASJo: Aceh Sanitation Journal, 1(1). Retrieved from
https://journal.poltekkesaceh.ac.id/index.php/asjo/article/view/104
Reyes, J. & dela Cruz, J. (2020). Repellency of Local Plants Against Mosquitoes.
Philippine Journal of Entomology, 34(1), 1-8.

Santos, J. E., Santos, J., Sulleza, D. M. S., & Tahanlangit, R. (2018, March).
PDF. Bulacan; La Consolacion University Philippines.

Samake, F. B., Traore, D., Diallo, D., & Coulibaly, A. (2011). Evaluation of
extracts from six medicinal plants from Mali as mosquito larvicides.
Journal of Insect Science, 11(72), 1-8. doi: 10.1673/031.011.7201

28
Shukla, D., Wijayapala, S., & Vankar, P. S. (2018). Effective mosquito
repellent from plant basedformulation. International Journal of Mosquito
Research, 5(1), 19-24.

Tantawichien T. (2015). DENGUE FEVER AND DENGUE HEMORRHAGIC

FEVER IN ADULTS. The Southeast Asian journal of tropical medicine and
public health, 46 Suppl 1,79–98.

Wang, X., & Hu, Y. (2018). Safety assessment of DEET and picaridin-based
insect repellents. Journal of toxicology, 2018.

World Health Organization. (2019). World Malaria Report 2019.

Xing, X., Liang, L., & Dong, H. (2018). Comparison of the repellency of plant
essential oils to mosquitoes (Diptera: Culicidae) under laboratory
conditions. Journal of Medical Entomology, 55(3), 587-593.

29