PASIG CITY SCIENCE HIGH SCHOOL

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Telefax No. 628-2177
S.Y. 2021-2022

The Scrutinization of the Durability of Oryza Sativa and Antimicrobial

Properties of Citrus Sinensis and Citrus Limon in the
Composition of Bioplastic Hand Gloves

Research Paper Submitted in Partial

Fulfillment of the Requirements
For Practical Research III

BANZIL, Jemima Keziah V.

BOLISAY, Nikita Ryle C.
CRUZ, Vladimir V.
SAGRADO, Ma. Michelle C.
TORRALBA, Xyrene Jade R.

10 – Goldstein

Mr. Filmar R. Domingo

2021 - 2022

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 TABLE OF CONTENTS

Chapter I The Problem and Its Background ............................................................................ 3

I. Introduction ............................................................................................................................ 3
II. Background of the Study....................................................................................................... 4
III. Statement of the Problem..................................................................................................... 6
IV. Hypothesis ............................................................................................................................ 9
V. Significance of the Study ...................................................................................................... 9
VI. Scope and Limitations of the Study..................................................................................... 9
VII. Definition of Terms .......................................................................................................... 11

Chapter II Review of Related Literature and Studies ........................................................... 17

I. Introductory .......................................................................................................................... 17
II. Related Literature ................................................................................................................ 17
Foreign Literatures ............................................................................................................... 18
Local Literatures .................................................................................................................. 19
III. Related Studies .................................................................................................................. 20
Foreign Studies .................................................................................................................... 20
Local Studies ........................................................................................................................ 21
IV. Synthesis ............................................................................................................................ 23
V. Methodological Framework ................................................................................................ 25

Chapter III Methodology of Research..................................................................................... 26

I. Research Method/s and Design/s ......................................................................................... 26
II. Population Frame and Sampling ......................................................................................... 28
III. Description of Subjects/Respondents ................................................................................ 29
IV. Research Instruments......................................................................................................... 29
V. Data Gathering Methods and Procedures ........................................................................... 30
VI. Statistical Treatment of Data (Quantitative Analysis of Data) .......................................... 37

Bibliography ................................................................................................................................ 41

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 Chapter I

The Problem and Its Background

I. Introduction

The hand is by chance the most complicated anatomical component in the human body. It

is often employed in daily activities because of its numerous roles. Humans rely on their hands

for both mechanical and sensory functions. As a result of the Covid-19 pandemic, many things

changed, including how we shop for our food and everyday essentials. Choosing the right glove

is crucial for avoiding cross-contamination and being in control of the situation while not posing

harm to the environment.

Traditional plastic, which is derived from petroleum, is difficult or impossible to recycle

and is made with chemical fillers that are harmful for the environment once melted down. As an

alternative, bioplastics are made from all-natural plant materials. Most bioplastics are expected to

minimize the use of fossil fuels, plastic waste, and carbon dioxide emissions. These polymers'

biodegradability has a beneficial influence on society, and awareness of biodegradable packaging

draws academics and industries. Plant-based plastics come in a variety of forms, including

essential oil, lemon and orange peels, starch, plants and others. Pure starch, on the other hand,

cannot be dissolved in cold water or alcohol. They have been utilized to manufacture

biodegradable films to partially or totally replace plastic polymers because of their low cost and

renewability, as well as their superior mechanical qualities.

Orange with a scientific name of Citrus Sinensis belongs to Rutaceae family and must

have originated between the borders of China and Vietnam. Citrus Sinensis is composed of peel

which is formed by flavedo and albedo, and the endocarp which is the pulp that contains vesicles

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with juice. 90% of the production is converted into juice in Brazil while it is consumed fresh in

South-East Asia. Molasses and pulp are being fed to cattles while essential oils and pectin are

made from the peel (Acedo et al. 2019). The fruit is at its best when it's freshly picked because it

contains no starch. The rind contains alkaloids, amino acids, carbohydrates, fixed oils and lipids,

proteins, steroids, sugars, tannins, and terpenoids.

Lemon with a scientific name of Citrus Limon also belong to Rutaceae family and was

introduced into Spain and North Africa first. The fruit itself is rich in vitamin C and contains

smaller amounts of B vitamins, particularly thiamin, riboflavin, and niacin. The outer shell, or

rind, is yellow when ripe, and in some varieties is quite thick and has prominent oil glands

embedded in it. The white, spongy interior of the shell, called mesocarp or albedo, has little taste

and is a major source of commercial pectin varieties. (Ganesh 2018). Thus, making the fruit’s by-

product of importance for other purposes.

II. Background of the Study

The world has a huge crisis regarding plastic, with the number one problem of polluting

bodies of the ocean and potentially wiping off an entire species. With the advancement of science,

we have made a possible solution to the ongoing crisis, and that is Bioplastics. Bioplastic is a

moldable plastic fabric made from chemical substances which might be derived from or

synthesized via means of microbes consisting of microorganisms or via way of means of

genetically changed plants. Unlike conventional plastics, which can be derived from petroleum,

bioplastics are acquired from renewable resources, and a few bioplastics are biodegradable. Due

to the negative environmental effects of synthetic plastics, the development of biodegradable

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polymers for both industrial and commercial applications is crucial today. Diverse starch-based

composites have been produced for various applications by researchers. The current study looks

at corn and rice starch-based bioplastics for protection purposes. Bioplastics of various

compositions concerning its durability, including corn and rice starch, glycerol, and vinegar are

created.

In citrus juices, there are numerous materials of various chemical nature, a number of

which own already-recognized hypocholesterolemia properties. Therefore, it has been regarded

exciting to assess the outcomes of extended management of lyophilized juices of Citrus sinensis

L. Osbeck and Citrus limon L. Burm fil. in diet-brought about rat hypercholesterolemia. Citruses

have a long-running history of different health benefits, but not just only health benefits, but also

anti-microbial fighting properties which can be further used for sanitary purposes. Citrus Sinensis

and Citrus Limon just to name a few examples. Earlier research of citrus oils from Citrus Sinensis

and Citrus Limon showed antibacterial activity against microbes. The oil extracts contain tannins,

saponins, flavonoids, terpenoids, cardiac glycosides, alkaloids, carbs (reducing sugar), and

phenol, according to phytochemical analysis. The highest concentration of flavonoids was found

in Citrus Sinensis. The presence of several functional groups in the extracted oils was discovered

using infrared spectroscopy. The extracted oils had an antibacterial impact on the test organisms,

which was attributed to the oil's phytochemicals and functional groups in antimicrobial tests. On

the organisms studied, the oils had synergistic antibacterial properties when used together. As a

result, we have a foundation for leveraging Starch-based bioplastic, Citrus Sinensis, Citrus Limon

to hook on to their efficacy in the production of antimicrobial bioplastic hand gloves.

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III. Statement of the Problem

The study will be conducted with the aim to show the ability of utilization and

manipulation of Oryza Sativa Starch in the production of Durable Bioplastic Hand Gloves with

Antimicrobial Properties. Citrus Sinensis and Citrus Limon Oil Extracts will both be utilized in

this study to observe the antimicrobial activity of the fruits and determine its effectivity in

reducing the growth of microbes. This study will utilize the bioplastics produced to perform the

two experimentations, the durability experimentation and microbial experimentation.

Specifically, it intends to answer the following:

a. How many among the experimental bioplastic compositions passed the pre-test of the non-

randomized pre-test posttest control group design intended for the durability

experimentation?

b. From the experimental bioplastic compositions that passed the pre-test, determine the

durability of the bioplastic by conducting the four sub-components of the durability test

and assessing the consistency of the data.

1. Leak Test

1.1 What is the average number of trials passed by Bioplastic Product 1?

1.2 What is the average number of trials passed by Bioplastic Product 2?

2. Elasticity Test

2.1 What is the mean of the level of the modulus of elasticity of the Bioplastic Product

1? Determine if the modulus of elasticity is high or low.

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 2.2 What is the mean of the level of the modulus of elasticity of the Bioplastic Product

2? Determine if the modulus of elasticity is high or low.

2.3 What is the mean of the level of the modulus of elasticity of the Bioplastic Product

3? Determine if the modulus of elasticity is high or low.

3. Durability Test and Degradation Test

3.1 Did the volume of Bioplastic Product 1 change or not? By what quantity?

a. After 1 hour?

b. After 3 hours?

c. After 6 hours?

3.2 Did the volume of Bioplastic Product 2 change or not? By what quantity?

a. After 1 hour?

b. After 3 hours?

c. After 6 hours?

3.3 Did the volume of Bioplastic Product 3 change or not? By what quantity?

a. After 1 hour?

b. After 3 hours?

c. After 6 hours?

4. Tensile Test

4.1 Will the Bioplastic Product 1 be resistant to:

a. 7.5g object?

b. 15g object?

c. 22.5g object?

4.2 Will the Bioplastic Product 2 be resistant to:

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 a. 7.5g object?

b. 15g object?

c. 22.5g object?

4.3 Will the Bioplastic Product 3 be resistant to:

a. 7.5g object?

b. 15g object?

c. 22.5g object?

c. Determine the difference in the growth of microorganisms through the comparison of the

results of the microbial test to be conducted with the bioplastic hand gloves produced with

Citrus Sinensis and Citrus Limon Oil Extracts and bioplastic hand gloves produced

without Citrus Sinensis and Citrus Limon Oil Extracts.

1. Microbial Test

1.1 Through observing the petri dish, determine how many large clusters (radius of

8cm) of microorganisms are observable in Petri Dish 1, 2, and 3 tested with

Bioplastic Hand Gloves with Citrus Sinensis and Citrus Limon Oil Extracts 1.

1.2 Through observing the petri dish, determine how many large clusters (radius of

8cm) of microorganisms are observable in Petri Dish 4, 5, and 6 tested with

Bioplastic Hand Gloves with Citrus Sinensis and Citrus Limon Oil Extracts 2.

1.3 Through observing the petri dish, determine how many large clusters (radius of

8cm) of microorganisms are observable in Petri Dish 7, 8, and 9 tested with

Bioplastic Hand Gloves without Citrus Sinensis and Citrus Limon Oil Extracts 1.

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 1.4 Through observing the petri dish, determine how many large clusters (radius of

8cm) of microorganisms are observable in Petri Dish 10, 11, and 12 tested with

Bioplastic Hand Gloves without Citrus Sinensis and Citrus Limon Oil Extracts 2.

IV. Hypothesis

Null Hypothesis

The researchers hypothesize that the utilization and manipulation of Oryza Sativa Starch

in the production of bioplastic hand gloves will not possess significant effects in making the

bioplastic durable. Additionally, the researchers hypothesize that the addition of Citrus Sinensis

and Citrus Limon Oil Extracts in the composition of the bioplastic hand gloves will not produce

a remarkable decrease in the growth of microorganism clusters.

V. Significance of the Study

The researchers will conduct this study with the aim to determine the durable properties

of Oryza Sativa Starch and assess the antimicrobial properties of Citrus Sinensis and Citrus Limon

Oil Extracts. The prototype bioplastic hand gloves will undergo two experimentations, the

durability experimentation and microbial experimentation. Through conducting these

experimentations, the researchers will be able to assess the effect of the utilization and

manipulation of Oryza Sativa Starch in producing durable bioplastic hand gloves. Additionally,

the researchers will be able to determine the difference between the growth of microorganisms in

bioplastic hand gloves produced with Citrus Sinensis and Citrus Limon Oil Extracts. In this study,

the researchers will produce Durable Bioplastic Hand Gloves with Antimicrobial Properties that

can be utilized in the day to day lives of selected people. This study will be beneficial to:

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STUDENTS

This study will be able to help students with similar studies in the future. The results of

the durability experimentation and microbial experimentation as well as the interpretation of the

gathered data will aid students in determining and assessing the properties of Oryza Sativa Starch

and the effect of its utilization and manipulation in the production of bioplastics. Additionally,

the students will be able to assess the antimicrobial properties of Citrus Sinensis and Citrus Limon

Oil Extracts in terms of decreasing the growth rate of microorganisms.

RESEARCHERS

This study will aid researchers with similar studies in the future as well. Through the

methods and experimentations performed by the researchers, future researchers can gather

relevant information crucial to their study. Furthermore, the future researchers can learn from the

methods conducted by the researchers in this study. The data gathering methods conducted by the

researchers will serve as a new reference for the future researchers. Lastly, through this study,

future researchers will be able to compare and contrast the information and data with other studies

which may aid in the improvement of the future researchers’ studies.

ENVIRONMENT

This study aims to produce durable bioplastic hand gloves with antimicrobial properties.

Plastics have been a problem for the environment as it could take 450 years or more before it can

fully decompose (Yu 2020). On the contrary, the duration for bioplastics to fully decompose only

take three to six months which is considerably faster than traditional plastics (BBC Science Focus

2022). Through the production of durable bioplastic hand gloves with antimicrobial properties,

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the utilization of traditional plastics in hand gloves can be decreased which may help in reducing

the plastic wastes that pollute the environment.

VI. Scope and Limitations of the Study

The intended bioplastic products will be created in each of the researchers’ houses. The

experimentation for the antimicrobial properties of the bioplastic hand glove will be safely

performed in an enclosed room without risking the safety of the researchers. Furthermore, the

experimentation that will be conducted to test the durability of the bioplastic hand glove will

happen on the respective homes of the researchers as well. In the span of three months, the

researchers shall be able to carry out a prototype of the product. The product is a sanitary hand

glove with antimicrobial properties intended to keep the users’ hands clean as they perform

activities outside their houses. The primary purpose of the bioplastic hand glove is to keep people

safe and protected from microorganisms that can be encountered outside while also being an eco-

friendlier option compared to existing hand gloves.

The scope and limitations are specified by researchers in this part of the study.

Scope

In conducting the research, the researchers considered the following:

1. The main component to be utilized in the production of the durable bioplastic hand glove is the

starch of Oryza Sativa. The rice starch will be used to make the hand glove through the process

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of molding. The researchers chose Oryza Sativa starch as the primary ingredient due to its

biodegradable properties that makes the hand glove eco-friendly.

2. The components with antimicrobial properties that will be used in the production of the hand

glove are the peels from Citrus Limon and Citrus Sinensis along with their extract to create the

oil extracts. The utilization of the oil extracts of these citrus products aid in making the bioplastic

hand glove have properties to decrease the growth of microorganisms.

3. The effectiveness of the antimicrobial effects of Citrus Limon and Citrus Sinensis peels and

extract are considered as well. A pre-assessment will be conducted with the goal of observing

the growth of microorganisms in the presence the orange and lemon peels to determine their

thorough effectiveness.

4. There will be tests to be conducted with the purpose of examining and assessing the

durability, strength, and tear-proof properties of the prototype gloves that will be created by the

researchers. Through this series of tests, the efficacy of using Oryza Sativa starch to the

durability of the bioplastic gloves will be determined.

5. The experimentation for the antimicrobial properties of the prototype hand gloves will be

conducted in a safe place inside the homes of the researchers. The experimentation area must be

appropriate for the performance of the test to ensure that the results will be exact and correct.

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6. There will be separate experimentations for the durability and the antimicrobial properties of

the prototype hand gloves made by the researchers. The factors for testing the antimicrobial

effects of the hand gloves are considered and noted in the experimentation. The standards for the

durability of hand gloves will be applied in the experimentation to confirm if the prototype gloves

are tough enough to be used.

7. The researchers considered performing tests that will be fit to be conducted inside ordinary

houses as an alternative to the tests that normally should be carried out in scientific laboratories.

8. The researchers will compare the prototype hand glove to vinyl gloves to determine the

advantages and disadvantages of choosing bioplastic gloves over vinyl gloves. Additionally, the

comparison will be conducted to recognize the possible weak points of the prototype bioplastic

hand gloves to be created by the researchers.

9. The researchers will make an alternative hand glove molder to help form the structure of the

bioplastic hand glove. Through this molder, the researchers expect to get the desired product

outcome as the prototype.

10. The duration for the production of the bioplastic hand glove will be five to ten days.

Bioplastics are usually obtained in two days, however, our bioplastic hand glove will need time

to mold. For the mold to be durable and to avoid the destruction of the prototype hand glove, it

will be kept in a secure room for seven to ten days.

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11. The instrument that will be used in the microbial experimentation will be a homemade petri

dish. The researchers gathered information regarding the production of petri dish at home as a

cheaper alternative to laboratory petri dishes.

12. The researchers considered that instead of utilizing a microscope, the naked eye will be used

instead in observing the growth of microorganism clusters in the homemade petri dish as

laboratory microscopes are not available for use and the researchers do not have the resources to

buy a laboratory microscope.

Limitation

In conducting the research, the researchers did not consider the following:

1. The utilization of industrial chemicals will not be considered for the purpose of promoting the

biodegradability of the prototype hand gloves. Doing this will also prevent the chemicals from

interfering with the antimicrobial properties of Citrus Limon and Citrus Sinensis.

2. The prototype bioplastic hand glove will not be tested against actual viruses due to the risk of

the procedure. Actual viruses will not be a part of the primary experimentation conducted in the

houses of the researchers to avoid the possible dangers possessed by viruses and ensure the safety

of the researchers.

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3. The experimentation for the antimicrobial properties will not be conducted inside laboratories

in accordance with the COVID-19 protocol. Due to this, the possibility of lacking essential

materials and tools inevitably exists.

4. The biodegradable properties of the bioplastic hand gloves will not be tested. Experimentations

concerning the biodegradability of the prototype hand glove will not be conducted due to the long

duration of the decomposition process of bioplastics that usually takes three to six months. The

time period for this type of experimentation does not line up with the intended two months of

complete production and testing for the prototype hand gloves.

5. The possible effects of the hand glove on different skin types will not be considered by the

researchers because it needs to undergo human experimentation. Alternatively, the researchers

will ensure that there will be no use of components that may cause allergic reactions for the

prototype glove to be user-friendly.

6. The other factors in the production of the bioplastic hand glove such as temperature and storage

will not be highly considered because the production and experimentation for the prototype glove

will only be conducted at the houses of the researchers.

7. The preservation of the anti-microbial effect of Citrus Limon and Citrus Sinensis will not be

monitored due to the lack of tools only available in scientific laboratories, specifically, the sterile

storages and bacteria growth monitoring tools.

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VII. Definition of Terms

In this section, the researchers will define terms that are crucial for comprehensible

reading. This section is dedicated to defining the terms that are essential to the study and for better

understanding for the readers.

Antimicrobial activity. A collective term for any active principles (agents) that prevent bacterial

development. It provides protection against bacteria and transmission of diseases.

Citrus Limon. A citrus that is rich in vitamin C and contains smaller amounts of B vitamins,

particularly thiamin, riboflavin, and niacin.

Citrus Limon Oil Extract. An oil extracted from the rind of Citrus Limon. It also possesses

antimicrobial activity.

Citrus Sinensis. A citrus that is a combination of pomelo and mandarin orange. It is rich in fiber,

calcium, and vitamin C and D.

Citrus Sinensis Oil Extract. An oil extracted from the rind of Citrus Sinensis by drying and with

the use of vodka.

Durable. Relates to the glove's longevity- its wear life based on person’s expectations up to the

point at which the person considers the product no longer suitable for the tasks or "worn out."

Modulus of Elasticity. The measure of elasticity of a material which is bioplastic.

Oryza Sativa. Commonly known as rice, it is a grain that is widely consumed and a great source

of fiber, magnesium, and other nutrients.

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Oryza Sativa Starch. A starch obtained from rice, Oryza sativa. It is a component in making the

bioplastic mixture.

Chapter II

Review of Related Literature and Studies

I. Introduction

This chapter includes literatures and studies, the generalizations and conclusions of the

references used, the methodologies of the study, and lastly, the synthesis of all the gathered

information. The review of related literature and studies would be beneficial to the researchers as

it allows the researchers to gather references for the study and make comparisons of the past

articles and studies for the improvement of the current study the researchers have. The contents

included in this chapter would be beneficial to the researchers by familiarizing pieces of

information that are relevant and similar to the present.

II. Related Literature

This section includes literatures divided into two categories, namely foreign and local

literature. The contents of this section include articles and journals that are beneficial to the

researchers as it serves as a guide which provides more crucial ideas and methods that can be

utilized by the researchers in their study.

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Foreign Literatures

The active chemical components of Citrus Limon Essential Oil contribute to its

therapeutic advantages, which give it the reputation of being an energizing, cleansing, and

purifying oil. It is perfect for use in cosmetics, aromatherapy, massage, and household cleaning

products to purify the air and surfaces because of these properties. Its anti-bacterial properties can

be used to disinfect not only wounds but other items. Citrus Limon Oil's astringent characteristics

revitalize dull complexions, making it great for restoring luster to weary or sagging skin. The high

vitamin content of lemon oil enhances immunity by enhancing the body's ability to battle harmful

microorganisms.

Citrus Sinensis oil has antibacterial and cleansing properties in addition to promoting

relaxation. Its antimicrobial properties make it ideal for application on any infected wound, as it

inhibits bacterial development and infection. It has the ability to not only create a warm

environment, but also to strengthen and resist the immune system, as well as to kill bacteria in the

air.

Plant-plastics have the advantage of not requiring pesticides or chemicals to develop.

Plant-based plastics can be made from agricultural waste or scraps, reducing the demand for

valuable oil resources while also reusing waste and benefiting from lower carbon emissions.

Bamboo fiber and wheat straw are utilized in products, and bamboo is one of the hardiest plants

on the planet. Without the use of pesticides, this plant flourishes and is rarely infected. In addition,

scientists have discovered that bamboo has a special bacteria called Bamboo Kun, which confers

antibacterial qualities to the bamboo, which are subsequently transferred to bioplastic. (Ritchie

2020).

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Local Literatures

Bioplastic is a consequential innovation that will have safe and environmentally friendly

alternatives to commercialized plastics. There are three types of bioplastics that decompose. The

first kind are all products of starch, every so often in mixture with vegetable oil or animal fats to

alternative for the polymers crafted from petroleum. The other two types of biodegradable plastics

are oxo biodegradable and hydro biodegradable. Oxo biodegradable plastics are normally crafted

from an oil derivative, however incorporate an additive, permitting them to break down with the

presence of oxygen to be consumed by microorganisms. Whilst Hydro-biodegradable plastics are

starch-based plastics that degrade with moisture (Cataquis, Angeles, and Dancel 2019).

Starch is regarded as one of the best biopolymers with high potential due to its facility to

biodegrade, the fact that it is renewable and abundant, and its low cost. 50% of existing available

bioplastics is made by Starch. Native starch is used to make most of the Starch-based bioplastics

which are either extracted or can also be blended with natural or synthetic molecules. Starch-

based bioplastics are facile to make and have a wide range of applications in packaging (Abellas

et al. 2021). Furthermore, gathered data shows that corn and rice starches have shown better

biodegradability than the subsisting plastic materials.

Cornstarch-Based Biodegradable Plastic helps lessen biological problems caused by non-

biodegradable plastics. Producing this requires two with four tablespoons of cornstarch, one cup

of water, two tablespoons of vinegar, and two tablespoons of glycerin to making the bioplastic

reliable in terms of elasticity, tear resistance, appearance, texture, and odor (Guevarra et al. 2020).

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III. Related Studies

This section includes studies divided into two categories, namely foreign and local studies.

The contents of this section include studies conducted by researchers before, both locally and

internationally, related to the researchers’ study. The chosen studies are beneficial to the

researchers as it serves as a guide which provides more crucial ideas and methods that can be

utilized by the researchers in their own study.

Foreign Studies

The study about corn and rice starch-based bio-plastics as alternative packaging materials,

showed that the samples prepared from corn and rice starches have better biodegradability than

the existing plastic materials. The maximum tensile strength of the bioplastic is found to be 12.5

MPa. Investigation of the hybridization of proposed starch materials with other biomaterials

would be an interesting scope for this research. Due to the obtained properties of bioplastic, it

would be interesting to prepare polybags using this bioplastic with assumed lower cost. From the

above test results, it can be concluded that bioplastics can be used as packing materials and can

be used as an alternative to LDPE and HDPE plastic bags. (Jowaid et al. 2019)

As of late plastic removal has arisen as a tireless and possible ecological issue on the planet.

The inappropriate removal of the plastics has brought about the passing of millions of creatures

and decrease in the fruitfulness status of the dirt by unloading plastic waste on both land and

ocean. The ecological concern has expanded on the plastic was te contamination, which has come

about into a quick advancement in the natural cordial item. Bio plastics are quite possibly the

most inventive natural amicable material enjoying benefit of lower carbon impression. The

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Bioplastics like PHA (Polyhydroxya lkanoates), PLA (Polylacticacid), PHB

(Polyhydroxybutyrates) are fabricated which are biodegradable with comparable usefulness like

that of regular plastics and can possibly lessen the reliance on petro - plastic that could be a climate

al risk. The current survey will feature a few parts of bio plastics, its creation, application, reusing,

and ongoing improvement in bio plastic. Each country frantically needs to create bio plastic

utilization and its legitimate waste administration for pollution. (Palaniswamay et al. 2020)

A number of bioplastics, including those made with PLA, are "compostable": if they

are exposed to enough heat, moisture, and hungry microbes, the microbes will break them down

into plant material, carbon dioxide, and water. Some environmentalists are concerned about

carbon dioxide. "This carbon dioxide gas is released back into the atmosphere," explains Richard

Wool, a chemical engineer at the University of Delaware in Newark. "Composting bioplastics

appears to be a misguided solution." Almost all compostable plastic ends up in a landfill rather

than being composted. Bioplastics, like other plastics, remain intact when buried in the oxygen-

free environment of landfills. However, some scientists are concerned that bioplastics will slowly

decompose over time, emitting methane (CH4), a greenhouse gas 20 times more potent than

carbon dioxide (Ko and Ra, 2015).

Local Studies

The antimicrobial activity of citrus fruit extracts: An in-vitro assay. The goal of the study

was to determine the antibacterial activity of citrus fruit extracts and to add to the body of evidence

supporting the use of these extracts to treat superficial bacterial and fungal skin disorders.

Calamansi pulp extract proved to be an efficient antibacterial against S. Aureus, P. Aeruginosa,

and C. Albicans in vitro. We'll have to wait and see if this has any dermatological implications.

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This study is pertinent to our research since we are dealing with citrus fruits to prove their

antimicrobial properties.

Combinatorial Bacteriostatic Activities of Garlic Bulb, Lemon Peel, and Orange Peel

Extracts Against Common Foodborne Bacteria in a Tomato-Based Pasta Meat Sauce. Prior to

determining antibacterial effects against relevant microbes, this study highlighted the need of

establishing the influence of plant extract supplementation, specifically Garlic Bulb, Lemon Peel,

and Orange Peel extracts, on the sensory attribute of a food system. As a result, the defined plant

extract supplementation regimens for food will be meaningful and can be employed directly in

the formulation and development of food products. Individual and combinatorial plant extract

inhibitory concentrations against foodborne pathogens often connected to illnesses were also

determined in this study. While the established MICo values were effective to suppress the

detected resistant bacteria in a real food matrix, more research is needed to increase the

supplemented product's consumer appeal. As a result, this study was able to characterize the

bacteriostatic activities of Aqueous Garlic bulb, Ethanolic lemon peel, and Ethanolic orange peel

extracts against Escherichia coli K-12, E. coli, Salmonella Enterica Serovars, Listeria

Monocytogenes, and Staphylococcus Aureus, which is strangely enough relevant to our study.

Starch-based Bioplastic Synthesized from Corn Starch and the Relationship Between

Density and Durability of Bioplastic. The primary objective of this study was to investigate the

link between bioplastic density ratios (measured in grams of corn starch per milliliter of glycerin)

and bioplastic durability. The goal of this study is to see if there is a significant relationship

between these two variables and how they influence each other because it will be a good

determinant of the bioplastic's quality. The data shows that the plastic density is considerably

(positively) connected to the bioplastic sample's maximal weight. This research aids us in

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considering various characteristics of the (Bio) plasticity of the materials we intend to utilize, as

well as the traits they must possess.

IV. Synthesis

Citrus Limon Essential Oil's active chemical components contribute to its ideal use in

household cleaning solutions for purifying the air and surfaces. Citrus Limon Essential Oil's

antibacterial characteristics can be utilized to disinfect not only wounds but also other objects and

surfaces as well. Citrus Limon Oil’s high vitamin content boosts immunity by increasing the

body's ability to fight harmful bacteria.

As for the Citrus Sinensis Essential Oil, it promotes relaxation while also having

antimicrobial and cleaning qualities. Its antibacterial characteristics make it excellent for use on

any infected wound, as it prevents the growth of bacteria and infection. It may not only generate

a warm environment, but also strengthen and resist the immune system and eliminate

microorganisms in the air.

The Citrus Limon and Citrus Sinensis Oil Extracts both have contents that promotes

antimicrobial properties that are recommended in cleaning purposes as it is effectively functional

in this field of use. These Oil Extracts can serve as homemade cleaning products that can reduce

the growth of microorganism found in surfaces and objects people encounter in their day to day

lives. As the Citrus Limon and Citrus Sinensis fruits are deemed to have antimicrobial properties,

the researchers found it appropriate to utilize the oil extracts produced by theses Citrus fruits in

the production of bioplastic hand gloves with the aim to protect the hands of hand glove users

from the growth of microorganisms.

23 | P a g e
 In the next literature, the bioplastic properties were discussed. Bioplastic is viewed as a

significant progress and development that will provide safe and environmentally acceptable

alternatives as compared to commercially available polymers. Bioplastics breakdown in three

different ways. The first category includes all starch-based products, which are occasionally

combined with vegetable oil or animal fats to replace petroleum-based polymers.

In this study, the researchers will utilize Oryza Sativa, commonly known as rice, to have

the starch in the composition of the bioplastics that will be produced by the researchers. The

researchers chose to utilize and manipulate the Oryza Sativa Starch alongside with the originally

used Corn Starch in the production of the bioplastic hand gloves to determine the effectivity and

possible ability of Oryza Sativa Starch in the field of bioplastics.

Furthermore, the researchers chose to utilize the Citrus Sinensis and Citrus Limon Oil

Extracts as the researchers identified the effectivity and strength of the antimicrobial properties

of these Citrus fruits through the literatures and studies gathered that supported the initial idea of

the researchers in using the concept of eco-friendly options without the use of chemicals for the

production of the bioplastic hand gloves with antimicrobial properties. This applies in the choice

of utilizing Oryza Sativa Starch in the hand glove composition.

To conclude, the literature and studies included in this chapter of the review of related

literature and studies are all beneficial and supportive to the researchers’ study in the form of aid

in the crucial information needed in the study. The methodologies and procedures used in the

previous studies of the researchers aided in the improvement of the researchers’ study because

the researchers were able to identify the weaknesses and strengths in the previous methodologies

conducted and performed in the previous related studies.

24 | P a g e
V. Methodological Framework

25 | P a g e
 Chapter III

Methodology of Research

In this chapter, the research methodology of this study is included and written out. In more

detail, this chapter outlines the research methods and designs, population frame and sampling,

description of subjects or respondents, methodology of the research, the research instruments,

experimentation methods, data gathering methods and procedures, the research process, statistical

treatment of data, and the quantitative data analysis.

I. Research Method/s and Design/s

As a quantitative experimental research study, the researchers will utilize two types of

research design for the two experimentations that will be conducted to obtain data required for

the study. The first research design that will be used by the researchers is the Non-randomized

Pretest-Posttest Control Group Design. Non-randomized Pretest-Posttest Control Group Design

is defined as a research design that assigns the population and sampled to treatment conditions

but does not use random assignment.1

The samples from the population of produced bioplastics will first undergo the pretest for

the durability experimentation. After the bioplastic compositions successfully passed the pretest,

1
“Nonrandomized Controlled Trial or Pretest Posttest Study with a Control Group.” n.d.
www.cebc4cw.org. https://www.cebc4cw.org/glossary/nonrandomized-controlled-trial-or-
pretest-posttest-study-with-a
controlgroup#:~:text=Nonrandomized%20Controlled%20Trial%2C%20or%20Pretest.

26 | P a g e
the researchers will conduct the durability experimentations and all of its four subcomponents.

Next, after the bioplastic products passed the durability experimentations, the researchers will

conduct the posttest and analyze which bioplastic composition is most fit to proceed to the

production of bioplastic hand gloves that will be used for the microbial experimentation.

As for the microbial experimentation, the research design that will be used by the

researchers will be Randomized Posttest-Only Control Group Design. Randomized Posttest-Only

Control Group Design is defined as a basic experimental design wherein the chosen samples are

randomly assigned to either undergo and be part of the experimental set-up or the control set-up.

The date will only be measured once the experimentation takes place in order to determine the

effect of the experimental set-up.2

The samples will undergo Randomized Posttest-Only Control Group Design in the

microbial experimentation. In the microbial experimentation, the population are the produced

bioplastic hand gloves. The chosen samples will be four pairs of bioplastic hand gloves. The first

two pairs of gloves are composited with Citrus Sinensis and Citrus Limon Oil Extracts, while the

third and fourth pairs are not composited with Citrus Sinensis and Citrus Limon Oil Extracts.

The microbial testing utilized the Randomized Posttest-Only Control Group Design as the

researchers will randomly choose from the produced bioplastic hand glove pairs with Citrus

Sinensis and Citrus Limon Oil Extracts. The researchers will also randomly choose from the

bioplastic hand glove pairs without the citrus oil extracts. These chosen pairs will undergo the

2
Choueiry, George. n.d. “Posttest-Only Control Group Design: An Introduction –
Quantifying Health.” https://quantifyinghealth.com/posttest-only-control-group-design/.

27 | P a g e
microbial experimentation. Then, the posttest will be conducted to assess the antimicrobial

properties of the Citrus Sinensis and Citrus Limon Oil Extracts utilized in the production of the

bioplastic hand gloves used in the microbial experimentation.

II. Population Frame and Sampling

The selected population are the bioplastic compositions. The researchers will utilize a

Non-randomized Pretest-Posttest Control Group Design to determine the most durable Bioplastic

among the 5 bioplastic compositions. The most durable bioplastic composition will be chosen to

undergo the production of bioplastic hand gloves. Three pairs of hand gloves will be utilized for

the Randomized Posttest Control Group Design to identify the effectiveness of Citrus Sinensis

and Citrus Limon Oil Extracts in terms of anti-microbial activity. After these experimentations,

the researchers will be able to determine the Final Bioplastic Hand Gloves.

28 | P a g e
III. Description of Subjects/Respondents

For the study on the production of bioplastic hand gloves, the researchers used Oryza

Sativa, Citrus Limon, and Citrus Sinensis, which are widely known as rice starch, lemon, and

orange, respectively. Bioplastics made from various forms of starch are becoming more popular

as a raw material for plastic products since they are not only biodegradable but also renewable.

As a result, Rice Starch (Oryza Sativa) is being considered as a potential source for the production

of bioplastics in this study. Citrus Limon Essential Oil's active substance components contribute

to the oil's medicinal properties, earning it the moniker "invigorating, cleaning, and purifying."

Its antibacterial characteristics make it ideal for use in household cleaning products to detoxify

the air and surfaces.

Lemon oil's high vitamin content boosts immunity by boosting the body's ability to fight

harmful bacteria. Antibacterial and cleaning effects are found in Citrus Sinensis oil. Its

antibacterial characteristics make it excellent for use on an infected wound, as it prevents the

growth of bacteria and infection. It can not only generate a warm environment, but it can also

build and resist the immune system as well as kill microorganisms in the air. The purpose of the

materials employed in this study was to demonstrate the product's durability and antibacterial

efficacy.

IV. Research Instruments

The research instrument used by the researchers in this study to gather data from microbial

testing is the petri dish. The petri dish is a shallow round glass that’s cylindrical in shape,

particularly targeting a microbe needed for the test. By swiping the gloves on the petri dish, the

researchers will investigate possible growth of microbes on the petri dish, whether it’s minimal

29 | P a g e
or plenty. Through the gathered data, the researchers will know if the prototype bioplastic anti-

microbial glove is effective.

V. Data Gathering Methods and Procedures

I. Durability Experimentation

The researchers will perform the durability experimentation to gather data regarding the

durability of the bioplastic products made from the components that passed the pre-test in the

research design of Non-randomized Pre-test Posttest Control Group Design. There are four sub-

components of the durability experimentation namely Leak Test, Elasticity Test, Tensile Test,

and Durability and Degradation Test.

1. Leak Test

The first durability test that will be conducted by the researchers is the leak test. The leak

test will be conducted to determine if the bioplastic produced contains holes and damages that

have a factor in the durability of the bioplastic gloves.

To conduct the leak test, the researchers will utilize three different amounts of water.

These amounts are 10 milliliters, 20 milliliters, and 30 milliliters. There will be three trials for

each amount of water. These amounts of water will then be moved throughout the whole

bioplastic made.

30 | P a g e
Bioplastic 1

Milliliter (mL) Trial 1 Trial 2 Trial 3

10 mL

20 mL

30 mL

Total: 9 Trials

Bioplastic 2

Milliliter (mL) Trial 1 Trial 2 Trial 3

10 mL

20 mL

30 mL

Total: 9 Trials

Through this leak test, the researchers aim to find out if there are holes in the bioplastic

product made. The bioplastic will then pass the leak test if there are no holes or zero holes found

in the bioplastic.

2. Elasticity Test

The second durability test that will be conducted by the researchers is the elasticity test.

The elasticity test will be conducted by the researchers to determine the modulus of elasticity of

the bioplastic. By conducting this test, the researchers can determine if the bioplastic has a high

modulus of elasticity or low modulus of elasticity.

31 | P a g e
 To conduct the elasticity test, the researchers will first cut out three pieces of 2x2 inches

bioplastics that were initially produced by the researchers. Then, the researchers will utilize two

pieces of 0.5 kilograms weights. These two pieces of weights will be put on either side of the 2x2

inches bioplastic. The researchers will stretch out the bioplastic 0.5 inches at a time. After each

stretch, the researchers will wait for 1 minute to see if the bioplastic resists or allows deformation.

There will be three trials per 2x2 inches piece of bioplastic. There will be a total of three trials for

the elasticity test.

[2x2 inch] Bioplastic 1

Inches (in) 0.5 inches 1 inch 1.5 inches

2x2 inches

[2x2 inch] Bioplastic 2

Inches (in) 0.5 inches 1 inch 1.5 inches

2x2 inches

[2x2 inch] Bioplastic 3

Inches (in) 0.5 inches 1 inch 1.5 inches

2x2 inches

Total: 3 Trials

Through this elasticity test, the researchers aim to find out the level of the modulus of

elasticity of the plastic. The information gained in this elasticity test can help the researchers

assess the durability of the bioplastic. The bioplastic will be declared rigid and with a high

32 | P a g e
modulus of elasticity if it resists deformation. However, the bioplastic will be declared flexible if

it allows deformation. The bioplastic will then pass the elasticity test once it is able to go through

the trials without tearing apart, regardless of whether it is declared rigid or flexible.

3. Tensile Test

The third durability test that will be conducted by the researchers is the tensile test. The

tensile test will be conducted to determine if the bioplastic produced by the researchers can handle

the weight of objects ranging from seven grams to twenty-two grams.

To conduct the tensile test, the researchers will first cut out three pieces of 1x1 inches

bioplastics that were initially produced by the researchers. These three pieces of 1x1 inches

bioplastics will be put around a circle binder. The researchers will also utilize one to three pieces

of five-peso coins put inside a light plastic container. Then, the researchers will test the resistance

of the bioplastic on one to three pieces of five-peso coins within one minute. There will be three

trials for each bioplastic product.

[1x1 inch] Bioplastic 1

Weight (g) Trial 1 Trial 2 Trial 3

7.5 grams

15 grams

22.5 grams

Total: 9 Trials

[1x1 inch] Bioplastic 2

Weight (g) Trial 1 Trial 2 Trial 3

7.5 grams

33 | P a g e
 15 grams

22.5 grams

Total: 9 Trials

[1x1 inch] Bioplastic 3

Weight (g) Trial 1 Trial 2 Trial 3

7.5 grams

15 grams

22.5 grams

Total: 9 Trials

Through this tensile test, the researchers aim to find out if the bioplastic products can

handle the weight of objects in one minute. The bioplastic will then pass the tensile test if each

bioplastic product passed the nine trials for the tensile testing.

4. Durability and Degradation Test

The last test that will be conducted by the researchers is the durability and degradation

test. The durability and degradation test will be conducted by the researchers to determine the

change in the measurement of the length and width of the bioplastic underwater with the pressure

of an object.

To conduct the durability and degradation test, the researchers will first cut out three

pieces of 3x3 inches bioplastics that were initially produced by the researchers. These three

pieces of 3x3 inches bioplastics will be put under 300 mL cold water with the pressure of one

medium-sized white egg. The researchers will measure the change in the length and width of the

flat bioplastics in the duration of six full hours.

34 | P a g e
[3x3 inch] Bioplastic 1

Hour 1 hour 2 hours 3 hours 4 hours 5 hours 6 hours

3x3 inches

[3x3 inch] Bioplastic 2

Hour 1 hour 2 hours 3 hours 4 hours 5 hours 6 hours

3x3 inches

[3x3 inch] Bioplastic 3

Hour 1 hour 2 hours 3 hours 4 hours 5 hours 6 hours

3x3 inches

Total: 3 Trials

Through this durability and degradation test, the researchers aim to find out the durability

of the bioplastic by measuring the change in measurement in the length and the width of the

bioplastic when it is subjected to the pressure of an object under the water. The bioplastic will

then pass the durability and degradation test if there isn’t a large change in the measurement of

the bioplastic from 3x3 inches within the duration of 6 hours.

II. Microbial Experimentation

The researchers will perform the microbial experimentation to gather data regarding the

antimicrobial properties of the bioplastic hand glove made from the bioplastic components that

passed the pre-test in the research design of Non-randomized Pre-test Posttest Control Group

Design. Along with the bioplastic component comes the Citrus Sinensis and Citrus Limon Oil

35 | P a g e
Extracts in the composition of the bioplastic hand gloves that will be utilized in the microbial

experimentation. There are four pairs of bioplastic hand gloves that will be used in this

experimentation. The first and second pair will be the Bioplastic Hand Gloves with Citrus

Sinensis and Citrus Limon Oil Extracts, while the third and fourth pair will be the Bioplastic Hand

Gloves without Citrus Sinensis and Citrus Limon Oil Extracts.

In conducting the microbial experimentation, the researchers aim to determine the

difference in the growth of microorganisms through the comparison of the results of the microbial

test to be conducted with the bioplastic hand gloves produced with Citrus Sinensis and Citrus

Limon Oil Extracts and bioplastic hand gloves produced without Citrus Sinensis and Citrus

Limon Oil Extracts.

To conduct the microbial experimentation, the researchers will need twelve pieces of petri

dishes. The first three pieces of petri dish will be utilized for the first pair of the Bioplastic Hand

Gloves with Citrus Sinensis and Citrus Limon Oil Extracts. For the second pair of the Bioplastic

Hand Gloves with Citrus Sinensis and Citrus Limon Oil Extracts, petri dishes 4, 5, and 6 will be

utilized. As for the first pair of the Bioplastic Hand Gloves without Citrus Sinensis and Citrus

Limon Oil Extracts, petri dishes 7, 8, and 9 will be used. Lastly, petri dishes 10, 11, and 12 will

be utilized for the second pair of the Bioplastic Hand Gloves without Citrus Sinensis and Citrus

Limon Oil Extracts.

III. Data Gathering and Interpretation of Results

The researchers will gather and tabulate all the results obtained from the tests. After

tabulating the results collected, the researchers will analyze and interpret the results. First, the

36 | P a g e
researchers will gather all of the data acquired from the durability experimentation. The

researchers will tabulate the results from the four sub-components of the durability testing namely

Leak Test, Elasticity Test, Tensile Test, and Durability and Degradation Test. After tabulating the

results, the researchers will statistically treat the data gathered from the Leak Test, Elasticity Test,

Tensile Test, and Durability and Degradation Test. Then, the researchers will gather all the data

acquired from the microbial experimentation. The researchers will tabulate the results from the

observation of the twelve pieces of petri dishes used in the microbial experimentation. After this,

the researchers will statistically treat the data obtained from the microbial experimentation. Next,

after the statistical treatment of data from the durability and microbial experimentation, the

researchers will use the necessary and appropriate graphs for the statistically treated data. Lastly,

the researchers will analyze and interpret the data and graphs and formulate a conclusion.

VI. Statistical Treatment of Data (Quantitative Analysis of Data)

Researchers will statistically analyze results collected from the observation with the data

necessities required in the study through determining the durability of the bioplastic by

conducting the four sub-components of the durability test and through observing the petri dish.

The goal of the durability test is to ensure that it is a necessary step in evaluating the bioplastic

composition's predicted lifespan. By recognizing faults at an early point in the bioplastic growth

process improves repeatability and decreases risk exposure. The data will be acquired from

observing the growth of mold from the homemade Petri dishes, then we will compare each Petri

dish and the mold that grew in it. It determines whether or not the gloves are sanitized and not

susceptible to bacteria and other known germs.

37 | P a g e
 The researchers will measure the Elasticity of the Bioplastic by Young’s modulus formula

which is Y = stress/strain. The ability of a material to endure variations in length when under

lengthwise tension or compression is measured by Young's modulus. This particular test will

cover the Initial Volume of Bioplastic, which corresponds to the measurement of the material to

be tested, as well as the Measure of Bioplastic after it has been stretched to a series of

measurements, which will lead to the determination of the Bioplastic's Elastic Modulus.

Y = Elastic Modulus

IV = Initial Volume of Bioplastic

M1 = Measure of Bioplastic after being stretched out to 0.5 inches

M2 = Measure of Bioplastic after being stretched out to 1 inch

M3 = Measure of Bioplastic after being stretched out to 1.5 inches

Y = stress/strain

Stress 1 = 0.5 inches

Stress 2 = 1 inch

Stress 3 = 1.5 inches

Strain 1 = IV - M1

Strain 2 = IV - M2

Strain 3 = IV - M3

The researchers will perform the Durability and Degradation test by using the formula TC

= IV - (C1 + C3 + C6). Given the circumstances, TC is equal to the quantity of the total change

38 | P a g e
in Volume, IV is equal to the Initial Volume of Bioplastic, and the C series will therefore be equal

to the change after 1 hour, 3 hours, and 6 hours respectively. Trying to figure out how long

bioplastic will survive and how well it will withstand cold water is difficult, but necessary.

Bioplastic's durability varies depending on where it's used, how long it's expected to last, and

what it's used for.

TC = Quantity of the total change in Volume

IV = Initial Volume of Bioplastic

C1 = Change After 1 hour

C3 = Change After 3 hours

C6 = Change After 6 hours

TC = IV - (C1 + C3 + C6)

In this test, the researchers will determine the difference in the growth of microorganisms

through the comparison of the results of the microbial test to be conducted with the bioplastic

hand gloves produced with Citrus Sinensis and Citrus Limon Oil Extracts and bioplastic hand

gloves produced without Citrus Sinensis and Citrus Limon Oil Extracts. The researchers will

calculate the Average Total of large Clusters (8cm in radius) in bioplastic hand gloves produced

with and without Citrus Sinensis and Citrus Limon Oil Extracts (ATW and ATWO) paying regard

to the number of Petri dish samples added (n) and Petri dish sample number (Pn).

ATW = Average Total of large Clusters (8cm in radius) in bioplastic hand gloves

produced with Citrus Sinensis and Citrus Limon Oil Extracts

39 | P a g e
 ATWO = Average Total of large Clusters (8cm in radius) in bioplastic hand gloves

produced without Citrus Sinensis and Citrus Limon Oil Extracts

n = Number of Petri dish samples added

Pn = Petri dish number

ATW = P1 + P2 + P3 + P4 + P5 + P6/n

ATWO = P7 + P8 + P9 + P10 + P11 + P12/n

Check to see which mixture produced the most massive clusters of microbes.

ATW > ATWO or ATW < ATWO or ATW = ATWO

If ATW > ATWO then the study is not efficient.

If ATW < ATWO then the study is efficient.

If ATW = ATWO then the study is neither efficient nor not.

40 | P a g e
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