Republic of the Philippines

Department of Education
REGION III – CENTRAL LUZON
SCHOOLS DIVISION OF BULACAN
SAN MIGUEL NATIONAL HIGH SCHOOL

A COMPARATIVE ANALYSIS: ANTIOXIDANT PROPERTY OF MANILA PALM

(Adonidia merrillii) FRUITS AND LEAVES USING MUNG BEAN (Vigna radiata)
MODEL

A Research Paper
Presented to the Faculty of Senior High School Department
San Miguel National High School
San Miguel, Bulacan

In Partial Fulfillment of the Requirements for the Subjects

Inquiries, Investigation and Immersion and Capstone Project

Patrick Allen A. Cruz

John Raven D. Del Rosario
Hanz Danniele M. Hirang
Chloe A. Santos
Lei Alvhie O. Soliman
Desiree Saira S. Varona
Researchers

Rachelle S. Viñas
Research Adviser

April 2021

San Miguel National High School

Scuala St., San Juan, San Miguel, Bulacan
Telephone Nos: (044) 327-1123 / (044) 327-1104
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INTRODUCTION

Free Radicals (FR) are substances that derive from incompletely oxidized compounds

that have undergone partial burning and that have oxygen groups capable of initiating aggressive

oxidation reactions.1 These substances come from both the environment (cigarette smoke,

polluted air, foods, medicines, well or tap water) and as by-products of chemical reactions of our

body systems like incomplete catabolism, energy production, hepatic detoxification, and more.

They bind with other molecules that cause several adverse effects that lead to the disruption of

normal processes. Macular degeneration, cardiovascular diseases, emphysema, Alzheimer’s

disease, Parkinson’s disease, ulcers, and inflammatory diseases such as arthritis and lupus are

just some diseases associated with free radicals.2

Alongside this harmful substance, the body also produces antioxidants to negate or

reduce their damage. Antioxidants are compounds capable of either delaying or inhibiting the

oxidation processes. They act as a donor of electron and hydrogen, radical scavenger, peroxide

decomposer, singlet oxygen quencher, enzyme inhibitor, synergist, and metal-chelating agents.3

In times that the ratio of production of antioxidants and free radicals becomes imbalanced,

oxidative stress occurs. It can damage many tissues in the body and eventually lead to a number

1
Monica Butnariu and Ionel Samfira, “Free Radicals and Oxidative Stress”, Journal of J Bioequivalence &
Bioavailability 4, no. 3 (April 2012): 1.
2
Jessie Szalay, “Live Science,” Accessed date April 30, 2021,
https://www.google.com/amp/s/www.livescience.com/amp/54901-free-radicals.html
3
Megan Ware, “Medical News Today”, Accessed Date April 30, 2021,
https://www.medicalnewstoday.com/articles/301506.
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of diseases over time.4 Due to such reason, many researchers have invested in finding sources of

antioxidants and assessing natural products’ antioxidant properties.

Manila palm (Adonidia merrillii (Becc)), or more commonly known as Bungang tsina is

a common ornamental plant native in the Philippines.5 More often than not, it is mistaken as its

more known relative, the arena nut palm which many locals use as a chewed stimulant alongside

betel leaf plant. Due to their similarities in both appearance and chemical profiling, several

studies were conducted to determine its potential in the field of medicine. One of which is its

antioxidant property. Its fruits showed high antioxidant activity compared to the control group

used.6 But so far, no study was done to test the antioxidant property of its leaves.

One of the ways to test the antioxidant capability of a plant extract is through the DPPH

assay. DPPH (1,1-diphenyl-2-picrylhydrazyl) is a stable free radical that can be reduced by

transferring a hydrogen from other compounds. It reduces when in the presence of an antioxidant

molecule, which forms into a colorless ethanol solution. Due to its simplicity and speed, it has

become the most commonly used method.7 But given the occurrence of the pandemic that

limited both resources and time, conducting such assay was made impossible. That is why a

plant model was used, specifically mung beans.

Mung bean (Vigna radiata) is one of the simplest ways to determine the antioxidant

potential of a plant. It involves the use of several setups to determine a plant extract’s antioxidant

4
Timothy J. Legg, “Health line: Everything You Should Know About Oxidative Stress,” Updated September 29,
2018, https://www.healthline.com/health/oxidative-stress.
5
“StuartxChange,” Philippine Medical Plants, Accessed date April 29, 2021,
stuartxchange.com/BungangDeJolo.html.
6
Ali Vafaei, Jamaludin Bin Mohamad, and Ehsan Karimi, “HPLC Profiling of Phenolics and Flavanoids of Adonia
merrillii Fruits and their antioxidant Properties,” Natural Product Research 33, no. 17 (2019): 2531-2535.
7
M.N. Alam, N.J. Bristi, and M. Rafiquzzaman, “Review on in vivo and in vitro methods evaluation of antioxidant
activity,” (n.p.: J. Saudi Pharm, 2013), 143–152.
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ability. Also, it does not require the use of dangerous chemicals, making it the most ideal method

for circumstances such as now.

As mentioned previously, there are no current studies that cover the antioxidant property

of Manila palm leaves. This study aims to evaluate and compare the antioxidant properties of

Manila palm fruits and leaves through the use of a mung bean model to determine which has a

higher capability to eradicate free radicals. The results will lead to a better understanding of the

plant’s constituents and provide an overview of which part of the plant should be the focus be on

in the production of antioxidants from natural sources.

Statement of the Problem

Manila palm is a common ornamental plant that can be seen in many landscapes. Studies

have been conducted to determine its medicinal potential. But given that different parts of a plant

can have varying properties, this study aims to determine which between the fruits and the leaves

of Manila palm have greater antioxidant property. Specifically, this study aims to answer the

following questions:

1. Does Manila palm (Adonidia Merrillii) fruit contain antioxidant property that

encourages the growth of the Mung beans (Vigna radiata)?

2. Do Manila palm (Adonidia Merrillii) leaves contain antioxidant property that

encourages the growth of the Mung beans (Vigna radiata)?

3. Is there a significant difference between the effectiveness of the Manila palm

(Adonidia Merrillii) fruit and leaves as antioxidant?

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4. Which among the treatments showed the greatest effectivity in increasing the growth

of the Mung beans (Vigna radiata)?

5. Is there a significant difference between the effectiveness of the Manila palm

(Adonidia Merrillii) fruit and the control group?

6. Is there a significant difference between the effectiveness of the Manila palm

(Adonidia Merrillii) leaves and the control group?

7. Are there significant differences among the effectiveness of the eight treatments?

Hypothesis

The following hypotheses were formulated:

1. The Manila palm (Adonidia merrillii) fruits do not contain anti-oxidant property that

encourages the growth of the Mung beans.

2. The Manila palm (Adonidia merrillii) leaves do not contain anti-oxidant property that

encourages the growth of the Mung beans.

3. There is no difference between the effectiveness of the Manila palm (Adonidia merrillii)

fruit and leaves as antioxidant.

4. There are no differences between the effectivity of the treatments.

5. There is no significant difference between the effectiveness of the Manila palm fruit and

the control group.

6. There is no significant difference between the effectiveness of the Manila palm leaves

and the control group.

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7. There are no significant differences among the effectiveness of the eight treatments.

Scope and Delimitation

This study focuses on determining which between the leaves and fruits of Manila palm

(Adonidia merrillii) contain a superior antioxidant property by using a mung bean plant model.

To determine the antioxidant capability of the two plant extract, eight treatments were used, two

of which served as the control group. This study was conducted in San Miguel National High

School in Scuala St., San Juan, San Miguel, Bulacan during the second semester, School Year

2020-2021.

This study is limited to comparing the two aforementioned plant extract. Due to the

pandemic, the researchers are unable to perform the experiments in a laboratory setting. With

limited resources that are available, improvisations were done.

Significance of the Study

This study aims to identify and evaluate the antioxidant property of Manila palm

(Adonidia merrillii) leaves and compare it to those of the plant’s fruits. Manila palm fruit has

already been proven to have flavonoids, a most common phytochemical in fruits and vegetables,

which plays an important role in the prevention of diseases related to oxidative stress. 8 The

importance of conducting this comparative study is to determine which is more effective of the

two mentioned parts. With the results, researchers can focus on developing antioxidant

8
Jovana Čvorović et al., “Chapter 22 – Bioavailability of Flavanoids: The Role of Cell Membrane Transforters,” in
Polyphenols: Mechanisms of Action in Human Health and Disease, second edition, ed. Ronald Ross Watson et al.
(Massachusetts: Science Direct, 2018), 295-320.
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medicines from the part that yields more effectivity. Diseases such as cancer, Parkinson’s,

Alzheimer’s, and atherosclerosis are just few degenerative diseases caused by oxidative stress

that can be prevented by plant-derived antioxidants.9 Furthermore, it is advantageous to Third

World nations such as the Philippines as it will possibly produce products that are more cost-

efficient than those of the synthetic ones.

Patients. Patients having diseases, mainly caused by free radicals from damaging cells

and DNA will benefit the most with the result of this study. This study will help them become

aware of which, between the Manila palm leaves and Manila palm fruits is a more effective

antioxidant to take. Therefore, providing them more benefits.

Community. This study could enhance the awareness and understanding of people

concerning antioxidant properties in Manila palm. Thus, people in the community like farmers

could produce more Manila palm trees, and pharmaceutical companies could manufacture

antioxidants supplements using the more effective antioxidant.

Researchers. This study could serve as a guide, in terms of methodology, for further

researches that are similar to this study.

Definition of Terms

Antioxidant. These refer to substances that can fight off free radicals, which cause harmful

effects to the body.

9
Aurelia Magdalena Pisoschi1 and Gheorghe Petre Negulescu, “Methods for Total Antioxidant Activity
Determination: A Review,” Biochem & Anal Biochem 1, no. 1 (2011): 1.
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Cancer. This disease develops when changes in the growth and division of genes occur and

spread throughout the body.

DPPH assay. It is an antioxidant assay that is commonly used by researchers but which we were

unable to due to the pandemic.

Flavonoids. These are phytochemicals found in fruits and vegetables that have great antioxidant

ability.

Free radicals. These are atoms with unpaired electrons which seek partners to bind with to

become stabilized. This process causes harmful effects to human cells.

Manila palm. The species of plant from which the fruits and leaves used in the analysis were

taken.

Mung bean plant model. This is the subject that was treated and whose growth was observed.

Oxidation. This refers to the loss of electrons from a substance during a reaction.

Oxidative stress. It is a condition caused by an imbalance between the antioxidant and free

radicals within the body. It can result to cell and tissue damage and contributes to the aging

process.
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METHODOLOGY

Process:

Preparation and
Gathering of Manila
extraction of Manila
Palm fruits and leaves
Palm fruits and leaves

Antioxidant Testing
using Mung Bean Gathering of results
Plant Model

Comparing the results

of the extracts and Conclusion
interpretation.

Figure 1. Research Process Diagram.

The study's entire approach is depicted in Figure 1. Manila palm fruits and leaves were

gathered from a residential area in San Miguel, Bulacan; then, a domesticated blender was used

to get the extracts from both parts. After being soaked in eight different treatments, the mung

beans were planted. The results were gathered afterward and then interpreted to measure the

difference between the two extracts.

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Preparation and Extraction of Manila Palm

Washing of the leaves

and fruits, twice in
Gathering of Manila Cutting of the washed
running water and
Palm fruits and leaves parts into tiny pieces
once using distilled
water

Transferring plant Filtration of leaves and Individual extraction

extracts in sepearte fruits using cheese of leaves and fruits
containers cloth using blender

Figure 2. Extraction of Manila Palm Fruits and Leaves Diagram.

Materials Apparatuses Equipment

Cheese Cloth Basin Domestic Blender

Distilled Water Containers

Knife

Manila palm fruits

Manila palm leaves

Scissors

Tap Water

Extraction of Manila Palm

Manila palm leaves and fruits were gathered from Obtinalla’s residence at Tartaro, San

Miguel, Bulacan, and washed in running water twice and once in distilled water. They were
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soaked in distilled water individually for three days. After that, they were then crushed using a

blender and the obtained mixtures were filtered using cheesecloth. Lastly, they were transferred

into different clean containers.

Antioxidant Test

Soaking the Mung

Preparation of eight Beans in the Planting the soaked
treatments tratements for 10 seeds
hours

Measuring the length

Recording the results of the seeds after
seven days

Figure 3. Antioxidant Test Methodology Diagram

Materials Apparatuses

Cheese Cloth Basin

Distilled Water Measuring Cup

Mung beans Ruler

Compost soil Stirring Rod

Seedling tray

Hydrogen peroxide

Vitamin C
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The procedures in making the treatments needed for this test are:

Treatment A. The seeds were soaked in 100 mL of distilled water for 10 hours. This

treatment will serve as the control.

Treatment B. Ten mL of hydrogen peroxide was added to 100 mL of distilled water.

Afterwards the seeds were soaked in the treatment for 10 hours. This will serve as the negative

control.

Treatment C. Add 10 mL of the plant extract (leaves) to 100 mL of distilled water.

Afterwards the seeds were soaked in the treatment for 10 hours.

Treatment D. Add 10 mL of the plant extract (fruits) to 100 mL of distilled water.

Afterwards the seeds were soaked in the treatment for 10 hours.

Treatment E. Add 250 mg vitamin C to 100 mL of distilled water. Afterwards the seeds

were soaked in the treatment for 10 hours.

Treatment F. Add 10 mL hydrogen peroxide (free-radical) and 10 mL plant extract

(leaves) to 100 mL of distilled water. Afterwards the seeds were soaked in the treatment for 10

hours.

Treatment G. Add 10 mL hydrogen peroxide (free-radical) and 10 mL plant extract

(fruits) to 100 mL of distilled water. Afterwards the seeds were soaked in the treatment for 10

hours.

Treatment H. Add 10 mL hydrogen peroxide and 250 mg vitamin C to 100 mL of

distilled water. Afterwards the seeds were soaked in the treatment for 10 hours.
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After soaking the seeds in the different treatments, they are planted into the seedling trays

containing compost soil. They are separated into eight set-ups according to the treatment they

were submerged in. For seven days, the mung beans were watered thrice a day using 300 mL of

distilled water per session. The length of the seedlings was then measured and recorded to be

interpreted using F-test or also known as ANOVA test. It was then further compared using

Scheffe test.

Waste Disposal

Excess chemicals used were disposed in its appropriate organic and inorganic waste

containers. Otherwise, chemicals can be disposed in soils. Solid and liquid wastes were collected

in a waste container and the disposed of.