As An Alternative Antibacterial Dishwashing Liquid - Suyat

OREGANO (Coleus amboinicus) LEAVES EXTRACT AS AN ALTERNATIVE
ANTIBACTERIAL DISHWASHING LIQUID
A Research Proposal Presented to the Senior High School Department

of Calasiao Comprehensive National High School
Calasiao, Pangasinan
In Partial Fulfillment of the Requirements for the Subjects

Inquiries, Investigations, and Immersion and Capstone Project
Herxie Yna T. Suyat

Benedict S. Pontawe
Mhariel Jea P. Cabalgada
Dhenielle Irish L. Gamboa
Adrian James A. Baldueza
Christine Edith B. Cancino
Hans Benedict M. Angud
Angel Shamer Barace
Erickson D. Cabatbat
Ginalyn F. Cabatbat
Princess A. Barogo
Carlos S. Arenas
July 2023
APPROVAL SHEET
This research entitled, “Oregano (Coleus amboinicus) Leaves Extract as an Alternative
Antibacterial Dishwashing Liquid,” prepared and submitted by Herxie Yna T. Suyat, Benedict S.
Pontawe, Mhariel Jea P. Cabalgada, Dhenielle Irish L. Gamboa, Adrian James A. Baldueza,
Christine Edith B. Cancino, Hans Benedict M. Angud, Angel Shamer Barace, Erickson D.
Cabatbat, Ginalyn F. Cabatbat, Princess A. Barogo, and Carlos S. Arenas in partial fulfillment of
the requirements in the subjects Inquiries, Investigations, and Immersion and Capstone Project
for Science, Technology, Engineering and Mathematics (STEM) has been examined and is
recommended for approval and acceptance for completion.
Approved on June 2, 2023, by the Panel for Oral Examination:
ENGR. MERIAN D. GALANG, PhD ENGR. JACKIELOUD D. DECENA

Panel Member Panel Member
IVANSHANE L. MALALA GENESIS S. PAREL

Panel Member Panel Member
Accepted by:
CHRISTOPHER V. ZARATE HERNANDO C. ABALOS, JR.

Inquiries, Investigations, and Capstone Project Adviser
Immersion Adviser
Approved:
DR. CHONA C. SAMSON, EdD

Principal IV
ACKNOWLEDGMENT
This research study would not have met its fulfillment without the unending motivation and
support towards this project. The researchers of this study give their utmost gratitude and
appreciation to the following people:
The researchers would like to thank and express their sincere gratitude to Almighty God
for giving them patience, perseverance, wisdom, will, protection, and knowledge to finish this
study.
To the researchers’ parents and relatives, who gave their financial and emotional support
and then guided, prayed, and motivated them in making this study, especially to the Suyat,
Gamboa, Barogo, and Barace Families, who did not hesitate to provide a place and needed
supplementary materials toward this project.
To the researchers’ classmates and friends, who provided them with oregano leaves, Joie
Andrei Dangoy and Brando Layda.
Lastly, the researchers would also like to thank their Practical Research II Adviser, Sir
Christopher Zarate, and their Work Immersion Adviser, Sir Hernando Abalos, along with Sir
Genesis Parel for guiding them in this study. They would also like to give special
acknowledgement to Ma’am Joan Daryl Abellara and Sir John Michael Panoy, who have been
patient and significant pillars in conducting the study.
Sincerely,
Herxie Yna T. Suyat, Angel Shamer Barace,
Benedict S. Pontawe, Ginalyn F. Cabatbat,
Mhariel Jea P. Cabalgada, Erickson D. Cabatbat,
Dhenielle Irish L. Gamboa, Hans Benedict M. Angud,
Princess A. Barogo, Carlos S. Arenas,
Christine Edith B. Cancino, Adrian James A. Baldueza,
ABSTRACT
Title: OREGANO (Coleous amboinicus) LEAVES EXTRACT AS AN ALTERNATIVE
ANTIBACTERIAL DISHWASHING LIQUID
Proponents: Herxie Yna T. Suyat, Benedict S. Pontawe, Mhariel Jea P. Cabalgada, Dhenielle
Irish L. Gamboa, Adrian James A. Baldueza, Christine Edith B. Cancino, Hans Benedict M.
Angud, Angel Shamer Barace, Erickson D. Cabatbat, Ginalyn F. Cabatbat, Princess A. Barogo,
Carlos S. Arenas
School: Calasiao Comprehensive National High School
In recent times, water is where pollution and disruption had been rampant in various
bodies. In addition, oregano and oregano extract both have significant antimicrobial content,
according to several test tube experiments, which are substances that protect the body from harm
caused by dangerous free radicals. This study aims to study the effectiveness of oregano leaf
extract as an alternative to dishwashing liquid.
In this study, the researchers carried out two (2) experiments. The Zone of Inhibition
test had three (3) replication and four (4) treatments; Positive control group, negative control
group, oregano extract, and oregano dishwashing liquid. And the dishwashing efficiency test had
three (3) replication and two (2) treatments, consisting of the experimental group (oregano
dishwashing liquid) and control group (commercial dishwashing liquid). The oregano dishwashing
liquid had undergone two standardized tests for antimicrobial activity and dishwashing efficiency.
The results indicated from the ANOVA and Independent T-test showed a significant difference in
the antibacterial activity with greater potent for sensitivity against the P. aeruginosa, while it had
shown potent dishwashing efficiency of the oregano extract’s capability of being a dishwashing
liquid.
TABLE OF CONTENTS
Title Page………………………………………………………………………………………….i
Approval Sheet…………………………………………………………………………………...ii
Acknowledgement……………………………………………………………………………….iii
Abstract………………………………………………………………………………...…………iv
CHAPTER I: THE PROBLEM
A. Background of the Study……………………………………………………………….1
B. Conceptual Framework………………………………………………………………….
C. Statement of the Problem…………………………………………………………….
D. Hypotheses of the Study…………………………………………………………….
E. Scope and Delimitations…………………………………………………………………
F. Significance of the Study………………………………………………………………
G. Definition of Terms………………………………………………………………………..
CHAPTER II: REVIEW OF RELATED LITERATURE AND OTHER STUDIES
A. Related Literature………………………………………………………………………..
B. Related Studies…………………………………………………………………………..
CHAPTER III: METHODOLOGY
A. Research Design………………………………………………………………………….
B. Procedures……………………………………………………………………………….
C. Data Gathering Process……………………………………………………………….
D. Instruments for Data Collection………………………………………………………..
E. Statistical Treatment of data……………………………………………………………
F. Ethical Conditions………………………………………………………………………
CHAPTER IV: RESULTS AND DISCUSSION
A. Presentation and Analysis of Reports………………………………………………….
CHAPTER V: SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS
A. Summary………………………………………………………………………………
B. Conclusions………………………………………………………………………………
C. Recommendations……………………………………………………………………
Bibliography……………………………………………………………………………………
Appendices……………………………………………………………………………………
Appendix A – Communication Letters………………………………………………
Appendix B – Tables……………………………………………………………………
Appendix C – Statistical Analysis……………………………………………………
Appendix D – Documentation…………………………………………………………
Curriculum Vitae………………………………………………………………………………
Chapter I
THE PROBLEM
Background of the Study
Conventional dishwashing liquids often contain synthetic chemicals and
surfactants that can be detrimental to aquatic ecosystems. According to the “Wastewater
treatment waster use” (2018), it comprises substances such as chemicals, oil, soap,
human waste, and food scraps. This includes water from washing machines, dishwashers,
toilets, showers, and bathtubs in houses. In addition, businesses and industries contribute
a portion of the used water that requires treatment. If wastewater is not adequately treated,
the ecosystem and human health could suffer. When these products are washed down
the drain, they can end up in water bodies, contributing to water pollution. This pollution
has far-reaching consequences, including the disruption of aquatic life, contamination of
drinking water sources, and ecological imbalance. Among these effects, include
impairment to fish and wildlife populations, depletion of oxygen, beach closures and other
limits on recreational water usage, limitations on the harvesting of fish and shellfish, and
water contamination.
In addition, Zachariah (2020), claims many commercial dishwashing liquids
contain chemicals such as phosphates, formaldehyde, chlorine, and synthetic fragrances,
which can have adverse effects on human health. In the environment, triclosan kills
beneficial bacteria, our body accumulates this substance, which leads to endocrine and
hormonal issues. Phosphates can cause long-term digestive problems, migraines, and
other health problems when consumed through our dishes that may not have been
properly rinsed. On the other hand, an article from Unilever entitled, “Triclosan and
triclocarban,” have stated that they replaced triclosan and triclocarban with a range of
alternatives, including natural and nature-inspired antibacterial ingredients across their
products.
Relatively, “Literature Oregano” (2016), states that oregano is a plant belonging to
the Labiatae botanical family, which includes many other herbs like rosemary, mint, basil,
sage, thyme, and lavender. It naturally grows in the warm tropical climates of Africa,
Australia, and Asia. It has natural phytochemical components, which are highly prized in
the pharmaceutical sector. Furthermore, Ajmera (2017), has uncovered some of its
outstanding potential advantages, such as its antioxidant content and antibacterial, anti-
cancer, and antioxidant effects. Carvacrol is the main component of oregano oil, which
kills germs with the advantages of aromaticity, safety, non-poison, high-efficiency
sterilization, and no compatibility contraindication. Oregano is an efficient antibacterial
ingredient in hand-washing and surface-cleaning detergent solutions and demonstrated
that it is a potential substitute for triclosan and chloroxylenol.
Despite the growing concern for the environment and the need for sustainable
alternatives, there is a lack of comprehensive research exploring natural substitutes for
conventional dishwashing liquids. While some studies have investigated the antimicrobial
properties of various plant extracts, including oregano, there is a need for more specific
research focusing on their application as effective dishwashing agents. By addressing this
literature gap, the study aims to contribute valuable insights into the potential of oregano
extract as an alternative antibacterial dishwashing liquid, providing a foundation for further
development and implementation of eco-friendly cleaning products.
To look into the usage of oregano extract and its properties by introducing an
oregano extract-based dishwashing liquid, which is expected to be biodegradable and
environmentally friendly, the study aims to reduce the pollution caused by conventional
detergents. Therefore, this had given the researchers the idea to study the antibacterial
activity of oregano as a substitute in dishwashing liquids.

Conceptual Framework
The figure below shows the key variables, independent, intervening, and dependent, in
relation to the study, “Oregano (Coleus amboinicus) Leaves Extract as an Alternative
Antibacterial Dishwashing Liquid.”
Dependent Variable
Intervening Variable
Independent Variable
Effectiveness of
Temperature,
Oregano Extract Oregano Dishwashing
Environment
Liquid
Figure 1. Research paradigm of the Study, “Oregano (Coleus amboinicus) Leaves

Extract as an Alternative Antibacterial Dishwashing Liquid”
The oregano (Coleus amboinicus) leaves extract acted as the independent
variable as it is composed of phytochemicals, carvacrol and thymol, that affect the
efficiency of the dishwashing liquid. The effectiveness of the oregano dishwashing liquid
was the dependent variable, relying on the varying presence of constituents of the oregano
extract in making the dishwashing liquid. In contrast, the temperature and the environment
were the intervening variables that cannot be controlled and might have changed the
independent variable's effect.

Statement of the Problem
This study aims to determine the effectiveness of oregano leaves extract as an
alternative antibacterial dishwashing liquid. Specifically, this study was conducted to
determine the following problems:
1. Is there any significant difference in the antibacterial activity of the treatment
groups, oregano leaves extract in dishwashing liquid, oregano extract, positive
control group, and negative control group, specifically, through the Zone of
Inhibition (ZOI) Test, against:
a. Staphylococcus aureus; and
b. Pseudomonas aeruginosa bacteria?
2. Is there a statistically significant difference between the dishwashing efficiency of
the oregano antibacterial dishwashing liquid to commercially available
dishwashing liquid in terms of:
a. Scrub test performance;
b. Foaming capacity; and
c. Rinsing capability of the dishwashing liquid?
Hypotheses
From the research objectives stated, the researchers tested the null hypotheses
at a 0.05 level of significance. The researchers also hypothesized to determine the
following findings:
1. There is no significant difference in the antibacterial activity of the treatment
groups, oregano leaves extract in dishwashing liquid, oregano extract, positive
control group, and negative control group, specifically, through the Zone of
Inhibition (ZOI) Test, against:

b. Pseudomonas aeruginosa bacteria.
2. There is no statistically significant difference between the dishwashing efficiency
of the oregano antibacterial dishwashing liquid to commercially available
dishwashing liquid in terms of:
c. Rinsing capability of the dishwashing liquid.
Scope and Delimitations
This study aimed to determine the effectiveness of oregano leaves extract as an
alternative antibacterial dishwashing liquid in terms of its antibacterial activity against
Staphylococcus aureus, and Pseudomonas aeruginosa, and its dishwashing efficiency
through its scrub test performance, foaming capacity, and rinsing capability.
The study took place at the Lyceum-Northwestern University, Dagupan City, for
the antibacterial activity test. While the dishwashing making and performance testing was
conducted at one of the researcher’s houses at the Municipality of Calasiao. The study
lasted from September 2022 up until July 2023.
Limitations that may have altered this study involve the researchers' gathering of
materials due to a limited budget and location. Another limitation that may altered this
study is the bounded time for the study. Because of the short time frame, the researchers
made use of already-grown oregano plants obtained from Dinalaoan, Calasiao,
Pangasinan.
Additionally, a few readily available pieces of equipment had influenced the study's
findings, growth and the environment where the researchers acquired the oregano might
also be at play that the researchers were not able to control. The researchers considered
the stated hypothesis conditions for the experiments in this study.
Significance of the Study
This research was made with the aim to provide crucial information and knowledge
regarding the chosen topic from the recent studies or theses, and related sites needed for
the expected importance and benefit of the following:
Environment. This research could help the environment. Dishwashing liquid that
is chemical-rich has many effects on our environment.
It can cause a serious type of algae to grow on the water's surface, obstructing
sunlight and causing the demise of marine life due to a lack of oxygen. Natural
compounds, like oregano, are non-toxic and biodegradable, making them an
environmentally benign substitute for the synthetic chemicals found in most commercial
dishwashing detergents. This can lessen the harm that dishwashing liquids does to the
environment, particularly to waterways.
Community. Dishwashing soap can benefit from the addition of oregano to it,
giving the community a safer and greener alternative to conventional detergents. Oregano
may also be a cost-effective and long-lasting solution for many households due to its
accessibility and affordability. In conclusion, including oregano in dish soap could
encourage a better way of life among the populace while also making dishwashing more
pleasurable and environmentally friendly.

Manufacturers. Manufacturers of oregano dishwashing liquid can benefit from the
option in providing alternatives instead of harmful chemicals such as profit from increased
sales and revenue due to the product's unique selling proposition, customer satisfaction,
and use of the natural oregano properties to combat stubborn grease and stains. By
investigating more oregano uses in the cleaning sector, it may also result in brand
recognition and maybe increase the product line.
Government. The government can benefit indirectly from the cleaning product
industry in several ways, including tax revenue, employment and economic growth,
regulation enforcement, consumer protection, and support for research and development.
The industry's growth is driven by factors such as rising disposable income, increased
awareness of soap and detergent benefits, and the demand for efficient and
environmentally friendly cleaning products.
Future Researchers. Further studies in this similar aspect and area of research
can benefit students, and contribute to a literature gap in bridging additional information
and reference.
Definition of Terms
The following are the terminologies used in this study to define operational and
conceptual to give the contextual interpretation.
Agar. Provides nutrients and a place for microorganisms to grow that is used for
testing. This had been used to culture the Staphyloccocus areus and Pseudomonas
aeruginosa in order to test the antibacterial activity of the oregano extract in dishwashing
liquid.
Antibacterial Activity. This inhibits the growth of microbes. It was measured to
determine the capability of the oregano dishwashing liquid.
Antibacterial Agent. This will eliminate microorganisms to measure the
effectiveness of the oregano to be an antibacterial agent. This had been the parameters
of the potency of oregano extract as an alternative dishwashing liquid.
Bacteria. Is a single-celled microbe that can cause some illnesses like food
poisoning. This microorganism will be cultured in agar and treated with the oregano
dishwashing liquid to test its antibacterial activity and efficiency.
Commercial. It is the marketed and established product consumed by the
population. This is one of the treatments used as a parameter standard to compare the
efficiency of the oregano dishwashing liquid.
Dishwashing liquid. It is a cleaning product used to clean tableware and similar
utensils. It is the type of cleaning product that will be formulated with oregano extract to
test its ability to effectively remove bacteria and dirt.
Extract. Is the diluted concentration commonly obtained from a plant to withdraw
its constituents. The concentration of which the oregano was obtained and used as the
antibacterial agent used in the study.

Health. The aforementioned inquiry will look into the success of the alternative in
obtaining and preserving physical wellness. This is one of the significances of the study,
promoting a healthier alternative to both the environment and the community.
Oregano. An herbal plant commonly used in the medicinal field to treat such as
respiratory illnesses. The oregano extract was obtained from the leaves of this plant and
used as an alternative antibacterial agent in dishwashing liquid.
Pathogen. A causative agent in the study, mainly agar-induced bacteria will be
used to test the antibacterial activity of the dishwashing liquid against.
Sensitivity Discs. These are five (5) mm round white filter papers used as vessels
in the Zone of Inhibition test to easily determine its inhibition within the agar plate. It had
been used in the study to determine the antibacterial activity of the four (4) treatments
against the Staphylococcus aureus and Pseudomonas aeruginosa bacteria.
Scrub. The act of rubbing the surface of the plate/s using a sponge to help
eliminate dirt. This had been one of the parameters in testing the oregano dishwashing
liquid and its dishwashing efficiency compared to the commercial dishwashing liquid in
measuring the number of the plates washed.
Foam. Group of bubbles that takes the dirt out of the plate. This had been one of
the parameters in testing the oregano dishwashing liquid and its dishwashing efficiency
compared to the commercial dishwashing liquid in its ability to foam.
Rinsing. To cleanse by flushing the foam with water. This had been one of the
parameters in testing the oregano dishwashing liquid and its dishwashing efficiency
compared to the commercial dishwashing liquid in measuring its amount of water to
remove residual foam.

Chapter II
REVIEW OF RELATED LITERATURE AND OTHER STUDIES
Related Literature
Oregano Uses and Chemical Properties
The fleshy leaves and stems of this erectly spreading plant have a strong aromatic
quality. Because of its potent antioxidant properties, the oregano plant is used as a herbal
medicine in countries like the Philippines (“Literature Oregano Research and Review
Paper,” 2016). According to “Herbal Medicine: Oregano” (2022), the anti-inflammatory,
anti-bacterial, anti-oxidant, anti-fungal, and anti-viral properties of oregano are due to a
rosmarinic acid compound, thymol, and carvacrol. Additionally, oregano includes sterols,
triterpenoids, flavonoids, vitamin C, and vitamin A. Recent studies have shown that it has
antibacterial properties that can be used to treat infections of the reproductive tracts,
making it the perfect medication for new mothers. The volatile oils in oregano and its
properties are thought to be in charge of reducing the risk of ingesting harmful bacteria,
parasites, and fungi by slowing the process of food spoilage. Vujicic et al. (2016), adds
asthma, bronchitis, cough, digestive issues (diarrhea, indigestion, stomach ache),
antibacterial and anti-inflammatory issues, menstrual disorders, and diabetes have all
been treated with oregano in traditional medicine.
According to Boudreaux (N.d.), phenols are organic substances composed of a
carbon atom in a benzene ring and a hydroxyl (OH) group, which cannot go through the
same oxidation events as alcohol. Teixeira et al. (2013), stated that the major components
of oregano essential oil were carvacrol, β‐fenchyl alcohol, thymol, and γ‐terpinene. Hot
water extract had the strongest antioxidant properties and the highest phenolic content.
All extracts were ineffective in inhibiting the growth of the seven tested bacteria. In
contrast, the essential oil inhibited the growth of all bacteria, causing greater reductions in
both Listeria strains (L. monocytogenes and L. innocua). The most important idea is that
carvacrol and thymol may be beneficial for managing bacterial growth in foods, as
demonstrated by a 2019 laboratory experiment (Brazier, 2020). Carvacrol and thymol
have been shown in numerous trials to be effective antibacterial agents against both
Gram-positive and Gram-negative bacteria. Although thymol and carvacrol have not yet
been converted as commercially accessible antibiotics, thymol has been included in the
formulation of many health products due to its antibacterial capabilities. Thymol and
carvacrol are typically regarded as safe to consume (Kachur & Suntres, 2020).
Oregano oil is a traditional disinfectant that is known to be effective against a range
of infections, including the respiratory system, as well as bacterial and viral conditions. It
is also known to have anti-inflammatory properties and to be effective at limiting the growth
of cancer cells. Although oregano oil is stronger than its dried counterpart, the latter still
contains numerous potent health benefits. Studies have shown that carvacrol, a phenol
antioxidant found in oregano, has impressive anti-inflammatory and antimicrobial
properties when taken as a dietary supplement or added to food (Jockers, 2022).
Negative Effects of Manufactured Dishwashing Liquid
Many dishwashing detergents are contaminated with hazardous chemicals.
Dishwashing detergents contain harmful ingredients that, if inhaled or consumed, can
cause adverse reactions. These ingredients have been linked to central nervous system
disorders and even sudden infant death syndrome. Chemicals like acetone,
benzaldehyde, benzyl acetate, and camphor can cause kidney damage, respiratory
irritation, confusion, and convulsions. Chlorine, carbamazepine, chlorophenyl, phenol, and
diethanolamine are dangerous chemicals in many dishwashing detergents. These
substances can be hazardous to people, pets, and other animals when ingested, and they
can also be dangerous when inhaled. As your lungs cannot remove toxins as effectively
as your stomach can, breathing in fumes containing these toxins can have negative effects
on your health (“Real Dangers of Chemical Dishwashing Detergents,” 2019).
Dishwashing soap may contain dangerous substances like preservatives,
fragrances, colorants, and surfactants that can irritate the skin and lungs, disrupt the
endocrine system, and even lead to cancer. Unknown fragrance ingredients may include
toxic pollutants and allergens. Surfactants can damage the environment and cause
contact dermatitis, while preservatives can be toxic to aquatic life. Unintentional
contaminants can also be discovered in the final product. When selecting dish detergents,
it's critical to consider the potential harm that these components may pose to the
environment and human health.
According to the article from Unilever entitled, “Triclosan and triclocarban,” they
have identified alternative safe and effective ingredients to respond to consumer’s
preferences. In 2015, they stopped manufacturing skin care and cleansing products with
triclosan and they committed to phase out triclosan and triclocarban across all products
by the end of 2018. Across their products, they have replaced triclosan and triclocarban
with a range of alternatives, including natural and nature-inspired antibacterial ingredients.
Determining Effectiveness of Dishwashing Liquid
According to “Dishwashing liquid online” (2020), aside from washing plates,
dishwashing liquid is effective for cleaning various surfaces such as countertops, window
sills, glass, and plastic. Dishwashing liquids can also be used to clean stainless steel
kitchen appliances to remove fingerprints, stains, and grease.
Chemical dishwashing liquids are made of various chemicals in different amounts
and for various uses.

Triclosan, Sodium Laureth Sulfate, Disodium, Formaldehyde, and Phosphates are
a few of the most prevalent base components in commercial chemical dishwashing
detergents. Despite the fact that these active compounds remove tenacious grease and
residue from utensils, they may pose environmental risks and health risks to people. A
common anti-bacterial and anti-fungal ingredient in dishwashing detergents is triclosan.
However, it is also known to wipe out helpful microbes in the environment. Also, when
triclosan builds up in the body, it might harm people's health. Sodium lauryl sulfate is
primarily utilized as a foaming agent to cut through grease and remove it from the surface.
Nevertheless, Sodium lauryl sulfate has the potential to penetrate skin cells and cause
rashes and allergies. Concentrated phosphorus is one of the main contributors to water
contamination. Since formaldehyde is a well-known carcinogen, the Food Safety and
Standards Authority of India (FSSAI) has prohibited its use in fish population preservation.
Artificial scents, coloring agents, and preservatives are all present in chemical
dishwashing solutions and are bad for human health.
Dishwashing liquids are given preservatives in order to prolong the product's shelf
life and prevent bacterial development and spoiling (“EWG’s guide to healthy cleaning,”
N.d.). Dishwashing liquids are given a nice scent by the addition of fragrances. However,
some scents can irritate persons with delicate skin or trigger allergic reactions in some
(“Environmental Protection Agency,” N.d). As a result of the foam, the cleaning agent can
remain on the surface longer and better penetrate caked-on dirt, grease, and oil. Grease
globules can still slide off the plates if a good wetting agent is present, even though foam
is essentially a detergent’s sidekick (Lowe, 2022).

Laboratory Methods in Studying Bacteria and Their Effects on Humans
Staphylococcus aureus
One of the most significant bacteria that affect humans and cause disease is
Staphylococcus aureus, which has long been acknowledged as such. It is the main
contributor to skin and soft tissue infections like cellulitis, furuncles, and abscesses (boils).
The majority of staph infections are not dangerous, but S. Serious infections like bone and
joint infections, pneumonia, and bloodstream infections can all be brought on by aureus
(“About Staphylococcus aureus - MN,” 2019).
Human epithelial keratinocytes (HEKa) exposed to S-derived substances
experienced in vitro experiments to investigate changes in gene expression profiles and
inflammatory response. biofilms or substances secreted by S. aureus that contain aureus.
Planktonic growth of aureus. Taken together, these findings support and enhance
previous findings indicating that S. Biofilms of Staphylococcus aureus contribute
significantly to the chronic inflammation associated with chronic wound infection (“Early
effects of Staphylococcus,” 2014).
For identification of S. aureus, the swab from swab rinsing solution was spread-
plated on mannitol salt agar (Oxoid) plates and incubated at 37°C for 24 hours. Using the
Staphylase Test Kit (Oxoid), colonies of Staphylococci that were coagulase positive were
verified. Gram-positive clustered cocci in yellow colonies that were oxidase-negative,
coagulase- and catalase-positive, and produced clots were identified as S. aureus and
kept for future research at 20 °C in 20% glycerol brain heart infusion broth (“Detection of
Pathogenic Escherichia coli,” 2015).

Pseudomonas aeruginosa
According to “Pseudomonas aeruginosa: Bacterium Risk Factors and
Symptoms” (2022), Pseudomonas aeruginosa is a type of bacteria that is typically present
in the environment, such as soil and water. Infected surfaces, hands, and medical
equipment can distribute it to patients in healthcare facilities. In certain people, it can lead
to serious infections. Some healthy individuals even have strains of it developing on their
skin in moist areas of their bodies, such as their armpits or genital region. This is referred
to as a pseudomonas infection.Pseudomonas infections can occur in many different ways.
It can grow on fruits and vegetables, so consuming contaminated food could make you ill.
Additionally, it flourishes in damp spaces like sinks, hot tubs, restrooms, and kitchens.
Hospitals are where the most serious infections happen. Humidifiers and unclean medical
equipment, such as catheters, are ideal environments for Pseudomonas aeruginosa
growth.
Healthcare professionals who don't properly wash their hands can also spread the
infection from an ill patient to you.
Wilson et al. (2022), stated in the article, “Pseudomonas aeruginosa,”
Pseudomonas aeruginosa is a gram-negative, aerobic, non-spore-producing rod that can
infect both immunocompetent and immunocompromised hosts with a range of diseases.
It is a very difficult organism to treat in modern medicine due to its propensity to infect
immunocompromised hosts, high flexibility, antibiotic resistance, and a wide spectrum of
dynamic defenses.
Additionally, Castro (2022), stated that, P. aeruginosa is disseminated through
poor hand hygiene, such as that of healthcare personnel, or through contaminated,
incompletely sterilized medical equipment. Bloodstream infections, pneumonia, urinary
tract infections, and surgical wound infections are among the common P. aeruginosa
infections associated with hospitals.

These infections often affect hospital patients, especially those with weaker
immune systems as a result of illnesses or prolonged treatments. If you are being treated
with a breathing machine, such as a mechanical ventilator, or other medical devices, such
as urinary or intravenous catheters, while hospitalized, your risk for a serious, life-
threatening P. aeruginosa infection increases. In healthy individuals, exposure to polluted
water can also result in mild P. aeruginosa infections.
MacConkey Agar and Mueller-Hinton Agar
MacConkey Agar
MacConkey agar not only favors Gram-negative species but also distinguishes them
from one another through lactose fermentation by preventing the growth of yeast and
Gram-positive bacteria, stated Audrey Wanger (2019), from the article, “MacConkey
Agar”.
The colonies of non-lactose fermenters are clear, whereas the colonies of lactose
ferments stain pink. MacConkey desired the ability to distinguish between several
intestinal organism types in addition to enriching Gram-negative bacteria. Determining if
a colony belonging to Escherichia coli (then Bacillus coli communis) or Salmonella
enterica serovar Typhi (then B. typhi abdominals) was of particular interest. Theodor
Escherich, after whom the genus Escherichia is named, had previously observed that E.
coli was a type of bacteria, and MacConkey used this information to help identify these
organisms even though further testing was necessary. Salmonella does not digest lactose,
whereas coli does, allowing for immediate visual confirmation or exclusion of these
organisms, according to Smith K. (2019), from the article, “The Origin of MacConkey
Agar.”
Mueller-Hinton Agar
Mueller and Hinton created it at first as a protein-free medium for the initial isolation
of Neisseria species. A standard solid medium called Mueller Hinton Agar is advised for
the research of bacterial susceptibility to antimicrobial drugs using the diffusion (Kirby-
Bauer method) or dilution in agar methods (“Mueller Hinton Agar: Composition,
Preparation, and Uses,” N.d.). For disc diffusion (also known as the Kirby Bauer method)
testing of antibiotic susceptibility, Mueller Hinton Agar was created specifically. For the
following reasons, it has been suggested by the CLSI as the best medium for testing an
antibiotic's susceptibility: It exhibits good reproducibility from batch to batch; It contains
little of certain inhibitors, such as trimethoprim, sulfonamide, and tetracycline; It promotes
the growth of both fastidious and non-fastidious bacterial pathogens; The starch in the
medium absorbs toxins, resulting in the reduction of their effects. It is a loose agar, which
allows for improved drug diffusion while keeping them from interacting with the antibiotics
(“Mueller Hinton Agar,” N.d.).
Alcoholic Solutions and Aqueous Solutions
According to Puyana et al. (2015), stated in the article, “Water as green extraction
solvent: Principles and Reasons for its Use”, water is considered a green solvent since it
is non-toxic to health and the environment. Additionally, it is also safer and less expensive
compared to other solvents such as alcohol. Using alcohol as a solvent in extracting
components of medicinal plants proved more potent than aqueous solutions. The
antibacterial efficacy of Tetrapleura tetraptera (T. tetraptera) that used ethanol as a solvent
for extraction is more effective than aqueous extracts (Enaregha et al. 2021; Gberikon et
al. 2015).
Dry Leaves and Fresh Leaves in Extracting Plant Components
According to Sieprawska et al. (2022), states in the article, “Modern Methods of
Pre-Treatment of Plant Material for the Extraction of Bioactive Compounds”, fresh tissue
extracts are used in extracting plant components in ethnomedicine, however, dried plants
are more commonly used when extracting plant components. This is because drying the
plant inhibits its metabolic processes rendering any changes in the plant’s chemical
composition. Ethnomedicine refers to traditional medical practices that are concerned with
the cultural interpretation of health and medicine. It also includes knowledge about the use
of natural pharmaceuticals and the ethnic group from which the same pharmacologically
active ingredients belong as well (Mahapatra et al., 2019.) Aqueous extracts of Eucalyptus
globulus leaves, according to Pinto et al. (2022), include metabolites, primarily
polyphenols, which are in charge of the plant's wide range of biological activities, including
antioxidant, antibacterial, and allelopathic characteristics. Extracts made from dried leaves
are known for their high concentrations of amino acid derivatives (“Untargeted
metabolomic profiling,” N.d.)
Clinical and Laboratory Standards Institute (CLSI) Guidelines and Zone of Inhibition
According to Weinstein & Lewis (2020), CLSI is a well-known organization
for creating Standards Development Organizations (SDO). It complies with every need for
the World Trade Organization's development of international standards, serves as the
Executive Secretariat for the committee of the International Organization for
Standardization, and is the only laboratory SDO recognized as a World Health
Organization collaborating center. For the past 50 years, CLSI has produced a library of
standards spanning the main areas of clinical laboratory medicine, totaling about 240.
There are more than 50 nations that use its measures.

As stated by Bhargav et al. (2016), the antibiotic's location is surrounded
by a zone of inhibition where bacterial colonies cannot flourish. You can gauge how
susceptible the bacteria are to an antibiotic by looking at the zone of inhibition. According
to the CLSI guidelines, an antibiotic is deemed susceptible if the diameter of the zone of
inhibition is greater than 21 millimeters. An antibiotic is deemed intermediate if it has a
diameter of 14-21 millimeters and it is resistant if it has 14 millimeters below.
Oregano Phytochemical Components
According to Timothy (2018), The aqueous extract of oregano was obtained. The
oregano herb extract was then used for preliminary phytochemical analysis.
Phytochemical analysis has revealed the presence of an increased concentration of
alkaloids, flavonoids, terpenoids, and carbohydrates. The total phenolic content of O.
vulgare was quantitatively measured to be 41 mg/g of the dry weight of extract expressed
as gallic acid equivalent. The total phenolic content was estimated. Oregano is a plant
with great potential and needs to be further explored by future researchers. The presence
of these phytochemicals indicates that O. vulgare may be used from a health perspective.
It could be a vital component for many medicines and drugs in the future.
According to this survey, varying ingredients and amounts can be found in
oregano essential oils and extracts, which can vary depending on the species, soil type,
climate, time of year, place of harvest, growth circumstances, and extraction method. Anti-
microbial properties of Oregano are Carvacrol, thymol, and non-volatile phenolic
substances including rosmarinic acid and flavones are two major types of phytochemicals
that are responsible for the development of oregano (Soltani, 2018).

Related Studies
There are various tests that a dishwashing liquid is needed to test the potency of
a product. Relatively, Axion, Surf, Mr. Clean, and Joy were the four commercial
dishwashing liquids that were tested by Cabalza (2016) to see if they had any antimicrobial
activity against the two test microorganisms, Escherichia coli, and Staphylococcus aureus.
Moreover, how well it removes grease and oil. It was stated the commercial dishwashing
liquid is more effective at killing off food-borne bacteria the larger the zone of inhibitions.
Furthermore, a larger zone of clearance suggests that grease and oil can be removed by
commercial dishwashing liquid. The antimicrobial susceptibility test is the aforementioned
microbiological test that the researcher employed. Cabalza tested their ability to remove
oil and grease in addition to their ability to eliminate foodborne bacteria and created rancid
oil using isolated microorganisms. The experiment's findings showed that the four
dishwashing liquids have the ability to remove oil and grease as well as kill food-borne
bacteria. Staphylococcus aureus was found to be easier to get rid of with the four
dishwashing liquids than Escherichia coli (Cabalza, 2016).
A test called the Assessment of Cleaning Performance determines how well a
liquid detergent formulation can emulsify oily stains. Included in this test is the direct
application of dishwashing detergent to a sponge. The quantity of plates washed is a
measure of performance, with a higher quantity suggesting better cleaning capabilities.
A test method used to gauge a dishwashing detergent's desired performance is
the Assessment of Foam Performance (volume and stability). The foam stability test
measures a dishwashing detergent's capacity to withstand foam depletion when exposed
to soil (Wearie, 2015).
In a study by Rhoades et al., they focused on testing oregano essential oil as an
antimicrobial additive to detergent for hand washing and food contact surface cleaning.
Wherein, the oregano essential oil (OEO) was combined in a liquid detergent solution. It
was then tested with the same procedures as a commercial antimicrobial soap through
handwashing trials of natural flora. The results have concurred that the OEO is an effective
additive antimicrobial in detergent solutions application in hand washing and surface
cleaning (Rhoades et al., 2013).
Additionally, according to Gnatta et al. (2021), one of the most significant
categories of household chemicals is hand dishwashing liquids (HDL). These are aqueous
solutions comprising surface-active ingredients and different additives, such as perfumes,
preservatives, antioxidants, sequestrants, pH stabilizers, colorants, and opacifiers that
give compositions a pearly look. Natural cleaning methods are currently being employed
since they may be safer and healthier for everyone, especially for the environment,
animals, and children. It may be simpler to use natural, non-toxic disinfectants.
Considering most plants found in kitchens or gardens have antibacterial qualities similar
to those found in commercial and synthetic chemical cleaners (Christiansen, 2018). To
combat bacteria that produce odors, antibacterial cleaning agents were initially launched
in the 1920s. Due to the U.S., Once the Food and Drug Administration (FDA) outlawed
the use of triclosan and the other two dozen chemicals frequently found in antibacterial
soap, well-known cleaning brands started employing conventional and natural
antibacterial substances as alternatives, as stated by Aguirre (2019).
Furthermore, a study by Lambino et al. (2022), found that one of the most
concerning ecological problems today is the spread of water hyacinth plants, which have
caused the loss of aquatic species in lakes, ponds, canals, and sewage systems. On the
other side, water hyacinth contains antibacterial qualities that help produce a variety of
goods.
Eichhornia crassipes, often known as water hyacinth, can be used to make dishwashing
solutions, thereby reducing the spread of the invasive plant. In light of this, the researchers
came to the following conclusions: Both the commercial product and the homemade
dishwashing liquid performed as effectively as dishwashers. Although the designed
product has a tiny advantage over the commercial product in terms of effectiveness, the
difference is too minor to be considered significant.
Both, however, are effective at preventing Staphylococcus aureus from
proliferating. As all of the experiments produced favorable results, it can be concluded that
water hyacinth has antibacterial properties and can be a key component of dishwashing
solutions.
SYNTHESIS
Oregano oil is a traditional disinfectant that is known to be effective against a
range of infections, including the respiratory system, as well as bacterial and viral
conditions. It is also known to have anti-inflammatory properties and to be effective at
limiting the growth of cancer cells. Because of its potent antioxidant properties, the
oregano plant is used as an herbal medicine in countries like the Philippines. The anti-
inflammatory, anti-bacterial, anti-oxidant, anti-fungal, and anti-viral properties of oregano
are due to a rosmarinic acid compound, thymol, and carvacrol. The most important idea
is that carvacrol and thymol may be beneficial for managing bacterial growth in foods, as
demonstrated by a 2019 laboratory experiment. The irony of modern living is that the
cleaning supplies we use to keep our homes clean somehow contribute to pollution and
environmental contamination. This has been made worse by wastewater or used water. If
wastewater is not adequately treated, the ecosystem and human health could suffer. In
the study of Unilever “Triclosan and Triclocarbon,” they have identified alternative safe
and effective ingredients to respond to consumer’s preference.

Across their products, they have replaced triclosan and triclocarban with a range
of alternatives, including natural and nature-inspired antibacterial ingredients. Many
dishwashing detergents are contaminated with hazardous chemicals. Dishwashing
detergents contain harmful ingredients that, if inhaled or consumed, can cause adverse
reactions. Hence, the need to further research on the potential of oregano as an alternative
antibacterial dishwashing liquid.

Chapter III
METHODOLOGY
This chapter focuses on collecting materials and pieces of equipment used, the
methodology and procedures followed, the statistical treatment, and the research design
within the study, “Oregano (Coleus amboinicus) Leaves Extract as Alternative
Antibacterial Dishwashing Liquid.”
Research Design
For this research, the nature of the study was specifically a true-experimental
design.
True-experimental research is an effective statistical method for determining a
cause-and-effect link between the variables involved and is one of the most accurate types
of study designs as it offers strong evidence to back up the existence of correlations
(Voxco, 2021). For this reason, this research used a post-test-only design in experiments
as this study aimed to test the efficacy of oregano as an alternative antibacterial
dishwashing liquid that will be based on the stated objectives. True-experimental research
helped in testing the hypotheses distinctively to determine the oregano’s antibacterial
capability to reduce the metabolic activity of pathogens (e.g., Staphylococcus aureus, and
Pseudomonas aeruginosa) through Zone of Inhibition (ZOI) test, and determined the
oregano dishwashing liquid’s scrub test performance, foaming capacity, and rinsing
capability. In order to exclude the potential influence of outside variables and guarantee
the generalizability of results, standardized procedures and randomized assignment of
variables were carried out in order to retrieve impartial and precise data results.
Procedures
In this experimental research, the procedures of making the dishwashing liquid out
of oregano and the process of data gathering were experimental. The methodology of this
study’s experiment was referenced with the research by Lambino et al. (2022),
“Antimicrobial Activity of Water Hyacinth (Eichhornia Crassipes) as a Dishwashing Liquid.”
1. Gathering of Materials and Equipment
The researchers have gathered the following materials for the experiment: metallic
basins, spoons, pails, pots, trays, plastic bottles and containers, sterile cotton swabs,
measuring cups, beakers, graduated cylinders, Erlenmeyer flasks, Petri dishes, filter
paper, sponges, and droppers. Other chemical ingredients, MacConkay Agar, Mueller-
Hinton Agar, sodium laureth sulfate (SLES), sodium chloride (Salt), cocamide
diethanolamine (Degreaser), colorant, ethylenediaminetetraacetic acid (EDTA; water
softener), were bought online as a dishwashing liquid kit.
In addition, the pieces of equipment utilized were an electronic weighing scale,
triple beam balance, an autoclave machine, an incubator, and a stove. To ensure
safety in conducting this experiment, the researchers were required to sterilization of
the aforementioned materials, use safety gloves, lab gowns, eye protection, a
facemask, and an apron, and practice precautions throughout this study.
2. Preparation of the Oregano (Coleus amboinicus)
The main ingredient of the study, oregano, was harvested by the researchers from
Calasiao, Pangasinan where an abundant patch of this plant was found and had been
permitted to make use in ample amounts.
“ 6 reasons to grow oregano” (2023), published that the oregano plant is
moderately invasive however, has proven to also benefit its own environments like
insects by habitability.
3. Obtaining Oregano (Coleus amboinicus) Extract Through Boiling Method
(Decoction)
The researchers will make use of the oregano leaves in obtaining their extracts
vital to the study. It was rinsed until any visible dirt was removed and had been dried
until the excess moisture was no longer observable. The fresh oregano leaves were
boiled for 20 minutes in 6 liters of water to obtain the aqueous extract of pure oregano
concentration. The leaves were transferred in another container through a strainer to
filter debris after a 10-minute cooldown.
4. Formulation of the Oregano (Coleus amboinicus) Dishwashing Liquid
After the decoction of the oregano, 4 liters of water in the aluminum bowl were
mixed with the ingredients from the dishwashing kit in the following order: SLES
400mL, Sodium chloride 232g, Degreaser 24mL, Foam booster 36mL, Colorant 8mL,
EDTA 12mL along with the 1.25L of oregano extracts. Because of their resistance to
corrosion and the fact that they don't leak chemicals when exposed to heat or sunlight,
aluminum wares often endure longer than glass or plastic (“MSU Extension,” 2022).
SLES was added to the bowl and stirred to dilute the substance before adding sodium
chloride. As it isn’t advisable to pour them all at once as it can affect the product’s
texture while being blended, the sodium chloride was added gradually. Following the
addition of the colorant, the degreaser was then poured over the mixture and agitated.
The EDTA was prepared by the researchers by pouring approximately 12mL of the
substance in and stirring for at least a minute. Lastly, the main ingredient was added
to the solution and was once more agitated for about three to five minutes. The mixture
was distributed in each 1-Liter bottle after the process and was left for 24 hours to rest.
5. Post-handling of Bacteria in Agar
A generous amount of disinfectant was poured into each petri dish and was left to soak
for 15 minutes to eliminate the remaining bacteria. The agar was then carefully scraped
into a plastic bag and disposed of properly at a known trash bin by the laboratory facilities.
Using an antibacterial dishwashing liquid and a sponge, the Petri dishes were washed
normally.
Data Gathering Process
A. Testing the Antibacterial Activities on the Agar-agar-induced bacteria of the
Dishwashing Liquid
The researchers tested the differences in the antibacterial activity between the
oregano dishwashing liquid and Brand X dishwashing liquid on the agar-induced
bacteria at the Lyceum-Northwestern University Biology laboratory.
1. Zone of Inhibition Test. The bacteria cultures of Staphylococcus aureus and
Pseudomonas aeruginosa were grown in the selective differential agar,
MacConkey, and later transferred to the Mueller-Hinton agar for the ZOI test
among the two (2) treatments. The bacteria’s susceptibility to the oregano
dishwashing liquid was observed based on the size of its inhibition zone. The
control group underwent the same procedure as the experimental, however,
with the treatment of the Brand X dishwashing liquid this time. There were three
(3) petri dishes for this test representing the three (3) replicates, and the zone
of inhibition was measured in millimeters using a vernier caliper.
a. Staphylococcus aureus bacteria
Table 1. Zone of Inhibition Test on Staphylococcus aureus
Trials Treatments
Treatment 1 Treatment 2
Positive Control Negative Treatment 4
Treatment 3
Group Control Oregano
Oregano
(Commercial Group Dishwashing
Extract
Dishwashing (Saline Liquid
Liquid) solution)
Trial 1 X X X X
Trial 2 X X X X
Trial 3 X X X X
b. Pseudomonas aeruginosa bacteria
Table 2. Zone of Inhibition Test on Pseudomonas aeruginosa
Trials Treatments
Treatment 3
Oregano
Extract
Liquid) solution)
Trial 1 X X X X
Trial 2 X X X X
Trial 3 X X X X
B. Testing the Dishwashing Efficiency of the Dishwashing Liquid
There were three (3) tests conducted by the researchers in order to examine
the dishwashing efficiency between the oregano dishwashing liquid and Brand X
dishwashing liquid. And, there were a total of three (3) trials throughout each test.
This had helped in determining the efficacy of the oregano dishwashing liquid as
a potent product for the standards.
1. Scrub test performance. In this test, the researchers based the cleaning
ability of each dishwashing liquid according to the number of plates that were
soaked in used oil, in its ability to wash them completely using 1 mL of each
dishwashing liquid.
Table 3. Scrub Test Performance
Trials Treatments
Number of Plates Washed
The experimental group Control group (1mL of
(1mL of oregano Brand X dishwashing
dishwashing liquid) liquid)
Trial 1 X X
Trial 2 X X
Trial 3 X X
2. Foaming Capacity. The foaming does not necessarily affect the cleaning
ability of the product (“Cleaner Solutions,” 2016), however, it helps in
maintaining a gentle way to clean surfaces (“Hygiene,” 2022). For this test,
10mL of both dishwashing liquids were each placed in a beaker and shaken
for a fixed amount of time. The foam from this step was measured in
centimeters.
Table 4. Foaming Capacity.
Treatments
Height of the Foam (cm)
Trial 1 X X
Trial 2 X X
Trial 3 X X
3. Rinsing capability. For this test, both dishwashing liquids were discharged
from their containers, leaving behind foam residual. The containers were filled
with 250mL of tap water and stirred until there is no more foam. This stirring
and draining process was repeated in order to test how easy it would be to
rinse each dishwashing liquid.
Table 5. Rinsing capability
Trials Treatments
Amount of Water Added to Residual Foam (mL)
Trial 1 X X
Trial 2 X X
Trial 3 X X
Instruments for Data Collection
The researchers made use of a ruler, vernier caliper and beaker to obtain the
following data:
In order to compare the foam performance of the oregano dishwashing liquid with
the Brand X dishwashing liquid, the height of the foam was measured in centimeters. The
higher the foam, the better the performance of the dishwashing liquid. The diameter of the
ZOI were measured and interpreted as indicated by the CLSI standard.

If the diameter is greater than 21 millimeters, the interpretation is sensitive; if the
diameter is equal to 14-20 millimeters, the interpretation is intermediate; and finally, if the
diameter is less than 14 millimeters, the interpretation is resistant. The amount of water in
rinsing capability was measured in liters and was interpreted with the lesser amount of
water needed, the greater it’s rinsing capability (Lambino et al., 2022).
Statistical Treatment of Data
In determining the antibacterial activity of the oregano extract in dishwashing liquid
to reduce the metabolic activity of agar-induced bacteria, the Zone Inhibition (ZOI) Test
was performed. The diameters were measured in millimeters and were interpreted as
follows:
Table 6. Representation for the average measurements of diameter basis for
susceptibility, intermediate, or resistant interpretation from the CLSI standard
Diameter (mm) Interpretation
> 21 millimeteres Sensitive
= 14-20 millimeters Intermediate
< 14 millimeters Resistant
The statistical tool used was the Analysis of Variance (ANOVA) to compare the
significant differences among the four (4) treatment groups in the first experiment, Zone
of Inhibition test. Moreover, the researcher had performed the post-hoc test to further
pinpoint which group shows this variance among the other three (3). The statistical
differences were determined at the 0.05 level of significance. If the p-value is less than
the alpha (0.05), and F-value is greater than F-critical, reject the null hypothesis;
Accept the statement that there is a significant difference in the antibacterial
activity of the treatment groups: oregano leaves extract in dishwashing liquid, oregano
extract, positive control group, and negative control group, specifically, through the Zone
of Inhibition (ZOI) Test, against Staphylococcus aureus and Pseudomonas aeruginosa
bacteria.
Independent t-test was used to compare the performance of the oregano
dishwashing liquid from the Brand X dishwashing liquid to determine its dishwashing
efficiency. These tests were (1) Scrub Test Performance, (2) Foaming Capacity, and (3)
Rinsing Capability. The Independent t-test, commonly known as the two-sample t-test, is
an inferential statistical test that assesses whether the means of two unrelated groups
differ statistically significantly from one another. It was used to analyze whether there was
a significant difference between oregano dishwashing liquid and Brand X dishwashing
liquid in terms of their dishwashing efficiency. If the p-value is less than the alpha, reject
the null hypothesis; accept the statement that there is a statistically significant difference
between the dishwashing efficiency of the oregano antibacterial dishwashing liquid to
commercially available dishwashing liquid in terms of scrub test performance, foaming
capacity, and rinsing capability of the dishwashing liquid. Microsoft Excel will be used in
this assessment and data analysis of the two groups.

Ethical Conditions
Ethics, as defined by the Belmont Report, encompasses three fundamental principles:
respect for persons, beneficence, and justice.
1. Risks Involved
In the formulation of the oregano dishwashing liquid, the chemical ingredients
involved are sodium laureth sulfate (SLES), cocamide diethanolamine (Degreaser),
colorant, ethylenediaminetetraacetic acid (EDTA; water softener). These concentrated
active ingredients can be harmful (Patel, 2020), and they can lead to irritation and
dryness of one’s skin (“Counter Conscious,” N.d.). Beneficence entails maximizing
benefits and minimizing harm to research participants. Researchers are obligated to
ensure that potential risks to participants are minimized through thoughtful study
design and rigorous risk-benefit assessments. Additionally, researchers must strive to
maximize the potential benefits that their research can bring to individuals and society
as a whole. As safety and precaution will be observed, the use of safety gloves, eye
protection, a facemask, and an apron will be practiced throughout this study.
2. Results Communication
The communication throughout should exhibit strong reliability, dependability, and
credibility. The researchers should employ impartiality, transparency, and honesty
about the data, findings, research methodologies, and status of publications. The
findings of the study must also be explained by the researchers in the light of their
best understanding. The information gathered by the researchers must, however, be
backed up by precise and transparent research records and sources. Finally, the
researchers must defend their findings and be willing to acknowledge mistakes in
following the first statement of results communication.

3. Post-Handling
All pathogens and harmful bacteria were discarded according to the laboratory
protocols and safety precautions as stated in the methodology procedures section 5.
These were all assisted by professional lab instructors, therefore, assuring the safety
and health of everyone that is and will be involved.

Chapter IV
RESULTS AND DISCUSSION
This chapter presents all the results and findings after the conducted experiments.
This research study tested if oregano (Coleus amboinicus) leaves extract can be an
alternative antibacterial dishwashing liquid. Specifically, it is sought to test if 1.) Is there
any significant difference in the antibacterial activity of the treatment groups, oregano
leaves extract in dishwashing liquid, commercial dishwashing liquid, positive control
group, and negative control group, specifically, through the Zone of Inhibition (ZOI) Test,
against Staphylococcus aureus; and Pseudomonas aeruginosa bacteria; and 2.) Is there
a statistically significant difference between the cleaning performance of the oregano
antibacterial dishwashing liquid to commercially available dishwashing liquid in terms of
scrub test performance, foaming capacity; and rinsing capability of the dishwashing liquid.
The evaluation was done in terms of the following:
1. Zone of Inhibition test
To determine the products’ antibacterial activity, the Zone of Inhibition test was
used. Four (4) treatments were used to compare their effectiveness against two (2)
bacteria cultures namely, Staphyloccocus areus and Pseudomonas aeruginosa. After the
impregnation of the sensitivity discs containing the four (4) treatments, three (3) replicates
were made. The researchers measured the inhibition zone in millimeters. Table 2.1. shows
the recorded inhibition zone on Staphylococcus aureus of all the treatments groups. The
table shows that Treatment 1 had 35.5mm for the first trial, 36mm for the second, and then
34 for the last. This pertains to the greatest zone of inhibition observed and means that it
had been potent as an antibacterial. Treatment 2 and 3 on the other hand, shows a
consistent 5mm for all trials. While Treatment 4 had 5mm in Trial 1 and 3, while an 8,5mm
inhibition zone in Trial 2. The latter treatment shows a potential in its antibacterial activity.
Table 7. Zone of Inhibition Test on Staphylococcus aureus
Trials Treatments
Treatment 3
Oregano
Extract
Liquid) solution)
Trial 1 35.5mm 5 mm 5 mm 5 mm
Trial 2 36 mm 5 mm 5 mm 8.5 mm
Trial 3 34 mm 5 mm 5 mm 5 mm
The Analysis of Variance (ANOVA) on the mean capacity of the zone of inhibition of
the treatments declared that there was a statistical significance among the three (3) trials
that was evident at 0.05 significance level. The data further analyzed implies that
Treatment 1 with 35.5mm, 36mm, and 34mm, has an average mean of 35.17mm.
Treatment 2 and 3 with a consistent 5mm has an average mean of 5mm. Lastly, Treatment
4 with 5mm, 8.5mm and 5mm has an average mean of 6.17mm. Treatment 1 (35.17mm)
> 21mm, meaning it had passed the CLSI standard as sensitive. The rest of the treatments
(5mm, 6.17mm) < 14mm shows that the bacteria is resistant in CLSI standard.
Table 8. The table shows the F-test result for determining the Zone of Inhibition on
Staphylococcus aureus (mm)
ANOVA
Source of Variation SS df F P-value F crit Decision
1.7217399205E-
Between Groups 1997.833333 3 515.5699 09 4.066181 Reject
Within Groups 10.33333333 8
Total 2008.166667 11
It is clearly stated in Table 8. that the P-value (0.0017) from the F-test result is lesser
than the alpha (0.05), and the F-value (515.569) is greater than the F crit (4.06618),
thus, the null hypothesis is rejected. Therefore, the oregano dishwashing liquid in terms
of the antibacterial activity, is resistant and had not met the CLSI standard.
Table 9. The table shows the post-hoc test result for the Zone of Inhibition on
Staphylococcus aureus (mm)
POST-HOC TEST (SA)

Groups P-value Significance
Treatment 1 v Treatment 2 5.24E-09 p<0.01
Treatment 2 v Treatment 3 6.0000 insignificant
Furthermore, a post-hoc test was done to determine where the differences truly lie
among the four (4) treatment groups. The result from the multiple comparison test shows
that all treatments compared to Treatment 1 had significant results, (5.24E-09, 5.2432E-
09, 7.17E-09) < 0.01, while Treatment 2 with Treatment 3 and 4 had insignificant results
(6.000, 1.4648) > 0.01, as well as Treatment 3 with Treatment 4 (1.46477) > 0.01.
Therefore, Treatment 1 with Treatment 2, Treatment 1 with Treatment 3, and Treatment
1 with Treatment 4, are where the differences lie.

Table 10. Zone of Inhibition Test on Pseudomonas aeruginosa
Trials Treatments
Treatment 3
Oregano
Extract
Liquid) solution)
Trial 1 36.5 5 5 10
Trial 2 33.5 5 5 8
Trial 3 33.5 5 8.5 5
The Analysis of Variance (ANOVA) on the mean capacity of the zone of inhibition of
the treatments declared that there was a statistical significance among the three (3) trials
Treatment 1 with 36.5mm, 33.5mm, and 33.5mm, has an average mean of 34.5mm.
Treatment 2 with a consistent 5mm has an average mean of 5mm. Then, Treatment 3 with
5mm, 5mm, and 8.5mm has an average mean of 6.17mm. Lastly, Treatment 4 with 10mm,
8mm and 5mm has an average mean of 7.67. Treatment 1 (34.5mm) > 21mm, meaning
it had passed the CLSI standard as sensitive. The rest of the treatments (5mm, 6.17mm,
7.67mm) < 14 shows that the bacteria is resistant in CLSI standard.
Table 11. The table shows the F-test result for determining the Zone of Inhibition on
Pseudomonas aeruginosa (mm)
ANOVA
Source of Variation SS df F P-value F crit Decision
1.13174E-
Between Groups 1802.833333 3 179.1636 07 4.066181 Reject
Within Groups 26.83333333 8
Total 1829.666667 11
It is clearly stated in Table 11. that the P-value (0.00021) from the F-test result is
lesser than the alpha (0.05), and the F-value (179.1636) is greater than the F crit
(4.06618), thus, the null hypothesis is rejected. Therefore, the oregano dishwashing
liquid in terms of the antibacterial activity, is resistant and had not met the CLSI
standard.
Table 12. The table shows the post-hoc result for determining the Zone of Inhibition on
Pseudomonas aeruginosa (mm)
POST-HOC TEST (PA)

Furthermore, a post-hoc test was done to determine where the differences truly lie
among the four (4) treatment groups. The result from the multiple comparison test shows
that all treatments compared to Treatment 1 had significant results, (2.72E-07, 3.74E-07,
5.72E-07) < 0.01, while Treatment 2 with Treatment 3 and 4 had insignificant results
(2.7464, 0.6743) > 0.01, as well as Treatment 3 with Treatment 4 (2.071105) > 0.01.
Therefore, Treatment 1 with Treatment 2, Treatment 1 with Treatment 3, and Treatment
1 with Treatment 4, are where the differences lie.

2. Testing the Dishwashing Efficiency of the Dishwashing Liquid
Table 13. Scrub Test Performance
Trials Treatments
(Number of Plates Washed)
Trial 1 3 3
Trial 2 3 3
Trial 3 3 3
The Independent t-test on the mean capacity of the scrub test performance of the
treatments declared that there was no statistical significance among the three (3) trials
Treatment 1 and Treatment 2 with a consistent 3 number of plates recorded per trial has
an average mean of 3 plates washed for both treatments.
Table 14. The table shows the t-test result for determining the scrub test performance
t-Test: Two-Sample Assuming Unequal Variances
t-value p- Decision
Groups Mean Variance value
Experimental Group 3 0 4.30265273 p>0.05 Accept
Control Group 3 0
It is clearly stated in Table 14. that the P-value from the t-test result is greater than
the alpha (0.05) thus, the null hypothesis is accepted.

Therefore, the oregano dishwashing liquid is as efficient and show no statistical
difference compared to the commercial dishwashing liquid, in terms of the scrub test
performance and number of plates washed.
Table 15. Foaming Capacity
Trials Treatments
(Height of the Foam)
Trial 1 1.5 cm 1 cm
Trial 2 3.2 cm 1 cm
Trial 3 0.3 cm 1.3 cm
The Independent t-test on the mean capacity of the foam capacity of the treatments
declared that there was no statistical significance among the three (3) trials that was
evident at 0.05 significance level. The data further analyzed implies that Treatment 1 with
1.5cm, 3.2cm, 0.3cm has a mean average of 2cm. While Treatment 2 with 1cm, 1cm,
1.3cm has a mean average of 1.10cm.
Table 16. shows the t-test result for determining the foaming capacity (cm)
Groups Mean Variance t-value p-value Decision

Experimental Group 2.00 1.09 4.30265273 0.2786 Accept
Control Group 1.10 0.03
It is clearly stated in Table 16. that the P-value (0.2786) from the t-test result is
greater than the alpha (0.05) thus, the null hypothesis is accepted.
difference compared to the commercial dishwashing liquid, in terms of the foaming
capacity by its height from the surface of the 10mL dishwashing liquid.
Table 17. Rinsing capability
Trials Treatments
(Amount of Water Added to Residual Foam)
Trial 1 500 mL 500 mL
The Independent t-Test on the mean capacity of the rinsing capability of the
treatments declared that there was no statistical significance among the three (3) trials
Treatment 1 with consistent 500mL has a mean average of 500mL. While Treatment 2
with 500mL, 250ml and 250mL has a mean average of 333.33mL.
Table 18. shows the t-test result for determining the foaming capacity (mL)
Groups Mean Variance t-value p-value Decision

Experimental Group 500.00 0 4.30265273 0.1835 Accept
Control Group 333.33 20833.33
It is clearly stated in Table 2.10 that the P-value (0.1835) from the t-test result is
greater than the alpha (0.05) thus, the null hypothesis is accepted.
difference compared to the commercial dishwashing liquid, in terms of the rinsing
capability and the amount of water needed to rinse residual foam on the plate. ANOVA
test and T-test assuming equal and unequal variances were statistically used with 0.05 as
its level of significance.

Chapter V
SUMMARY, CONCLUSIONS AND RECOMMENDATIONS
Summary
This study had focused on problems revolving around water, where pollution and
disruption had been rampant in various bodies, health, and literature gap. In addition,
oregano and oregano extract both have significant antimicrobial content, according to
several test tube experiments, which are substances that protect the body from harm
caused by dangerous free radicals. This study aims to study the effectiveness of oregano
leaf extract as an alternative to dishwashing liquid.
In this study, the researchers carried out two (2) experiments. The Zone of Inhibition
test had three (3) replication and four (4) treatments; Positive control group, negative
control group, oregano extract, and oregano dishwashing liquid. And the dishwashing
efficiency test had three (3) replication and two (2) treatments, consisting of the
experimental group (oregano dishwashing liquid) and control group (commercial
dishwashing liquid). The oregano dishwashing liquid had undergone two standardized
tests for antimicrobial activity and dishwashing efficiency. The results from the ANOVA
and Independent T-test showed a significant difference in the antibacterial activity with
greater potent for sensitivity against the P. aeruginosa, while it had shown potent
dishwashing efficiency of the oregano extract’s capability of being a dishwashing liquid.
Conclusions
1. There is a significant difference in the antibacterial activity of the treatment groups,
oregano leaves extract in dishwashing liquid, oregano extract, positive control
group, and negative control group, specifically, through the Zone of Inhibition (ZOI)
Test, against:

b. Pseudomonas aeruginosa bacteria.
The oregano dishwashing liquid is not as effective as the commercial dishwashing liquid
in terms of its antibacterial activity as the zone of inhibition for both bacteria (6.17, 7.67)
was less than 14mm, in terms of CLSI standard, meaning it is not yet susceptible to the
product.
2. The oregano dishwashing liquid was as effective as the commercial dishwashing
liquid in its dishwashing efficiency showing no statistical difference in terms of:
c. Rinsing capability of the dishwashing liquid.
Recommendations
The researchers recommend to focus on the significance of the amount of
concentration of oregano extract to the dishwashing liquid, growing oregano in a controlled
environment, and the alternative use of pure extract instead of decoction should also be
studied. To also include, the researchers recommend the use of dried leaves in the
oregano extract instead of fresh leaves. Another scope for the antibacterial aspect is to
test the extract on other common bacteria such as Escherichia coli. Performing a
phytochemical screening and comparison between the components and efficacy of
aqueous solutions and alcohol solutions would also be advised. Next, the dishwashing
efficiency testing should also be supervised by professionals. And to further innovate other
aspects, a study on the antioxidant and the anti-fungal properties of oregano should be
taken into consideration. Additionally, a deeper analysis on oregano phytochemicals with
regards to its antibacterial capability is also recommended. Lastly, an extensive
experimentation and observation should be emphasized.

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Zachariah, S. S. (2020, February 25). Check your labels: 5 harmful chemicals in
dishwash liquid damaging your skin. The Better India. Retrieved from
https://www.thebetterindia.com/216941/dishwash-liquid-toxins-skin-problems-drying-
itching-natural-solution-buy-online-lifestyle-ser106/
Ziff, A. (2022, September 20). Made safe viewpoint: A breakthrough in liquid

dishwashing detergent. MADE SAFE a program of Nontoxic Certified. Retrieved from
https://madesafe.org/blogs/viewpoint/breakthrough-liquid-dishwashing-
detergent#:~:text=Common%20dish%20detergent%20can%20contain,irritation%2C%20
cancer%20and%20endocrine%20disruption
APPENDIX “A”
Communication Letters
Chona C. Samson, Ed. D.

Principal IV
Calasiao Comprehensive National High School
Dear Madam:
Greetings!
The undersigned students are currently conducting a research entitled “OREGANO

(Coleus amboinicus) LEAVES EXTRACT AS AN ALTERNATIVE ANTIBACTERIAL
DISHWASHING LIQUID”, to determine whether oregano has a significant effect as an
alternative antibacterial dishwashing liquid and its cleaning performance against
commercially available dishwashing liquid.
In this regard, the researchers would like to ask for permission to allow the said
study to consult information outside the school premises to consult information with
regards to their methodology needed for their research. Rest assured that the research
will be conducted with utmost responsibility and confidentiality.
Your favorable action and support regarding this matter is highly appreciated.
Thank you and God Bless!
Very truly yours,

Benedict S. Pontawe
Herxie Yna T. Suyat
Recommending Approval:
CHRISTOPHER V. ZARATE
Research Adviser
Approved:
_____________ (signed)
CHONA C. SAMSON, Ed. D.
Principal IV
APPENDIX “B”
Tables
Table A.1: Zone of Inhibition Test on Staphylococcus aureus
Trials Treatments
Treatment 3
Oregano
Extract
Liquid) solution)
Trial 1 35.5 mm 5 mm 5 mm 5 mm
Trial 2 36 mm 5 mm 5 mm 8.5 mm
Trial 3 34 mm 5 mm 5 mm 5 mm
Table A.2: Zone of Inhibition Test on Pseudomonas aeruginosa
Trials Treatments
Treatment 3
Oregano
Extract
Liquid) solution)
Trial 3 33.5 mm 5 mm 8.5 mm 5 mm

Table A.3: Scrub Test Performance
Trials Treatments
(Number of Plates Washed)
Trial 1 3 3
Trial 2 3 3
Trial 3 3 3
Table A.4: Foaming Capacity
Trials Treatments
(Height of the Foam)
Trial 1 1.5 cm 1 cm
Trial 2 3.2 cm 1 cm
Trial 3 0.3 cm 1.3 cm

Table A.6: Rinsing Capability
Trials Treatments
(Amount of Water Added to Residual Foam)
Table A.7: Representation for the average measurements of diameter basis for
susceptibility, intermediate, or resistant interpretation from the CLSI standard
Diameter (mm) Interpretation
> 21 millimeteres Sensitive
= 14-20 millimeters Intermediate
< 14 millimeters Resistant

APPENDIX “C”
Statistical Analysis
Statistical Computation B.1: The computation using Analysis of Variance in terms of the
antibacterial capability of the four treatments: Positive control group (T1), Negative control
group (T2), Oregano extract (T3), and Oregano dishwashing liquid (T4) against
Staphylococcus aureus
SUMMARY
Groups Count Sum Average Variance
T1 3 105.5 35.16667 1.083333
T2 3 15 5 0
T3 3 15 5 0
T4 3 18.5 6.166667 4.083333
ANOVA
Source of
Variation SS df MS F P-value F crit
Between Groups 1997.833 3 665.9444 515.5699 1.7217399205E-09 4.066181
Within Groups 10.33333 8 1.291667
Total 2008.167 11
Statistical Computation B.2: The computation using Analysis of Variance in terms of the
Pseudomonas aeruginosa.
SUMMARY
Groups Count Sum Average Variance
T1 3 103.5 34.5 3
T2 3 15 5 0
T3 3 18.5 6.166667 4.083333
T4 3 23 7.666667 6.333333
ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups 1802.833 3 600.9444 179.1636 1.13174E-07 4.066181
Within Groups 26.83333 8 3.354167
Total 1829.667 11
Statistical Computation B.3: The computation using post-HOC test in terms of the
Staphylococcus aureus.
POST-HOC TEST (SA)

Treatment 1 v Treatment 3 5.2432e-09 p<0.01

Statistical Computation B.4: The computation using post-HOC test in terms of the
Pseudomonas aeruginosa.
POST-HOC TEST (PA)

Statistical Computation B.5: The computation using a T-test assuming equal variances in
terms of the scrub test performance of the oregano dishwashing liquid (Experimental
Group) against the commercial dishwashing liquid (Control Group)
t-Test: Two-Sample Assuming Equal Variances

Experimental Group Control
Group
Mean 3 3
Variance 0 0
Observations 2 2
Pooled Variance 0
Hypothesized Mean 0
Difference
df 2
t Stat 65535
P(T<=t) one-tail p>0.05
t Critical one-tail 2.91998558
P(T<=t) two-tail p>0.05
t Critical two-tail 4.30265273
Statistical Computation B.6: The computation using a T-test assuming unequal variances
in terms of the foaming capacity of the oregano dishwashing liquid (Experimental Group)
against the commercial dishwashing liquid (Control Group)

Experimental Group Control Group
Mean 2.00 1.10
Variance 1.09 0.03
Observations 3.00 3.00
Hypothesized Mean 0.00
Difference
df 2.00
t Stat 1.472970759
P(T<=t) one-tail 0.139326113
P(T<=t) two-tail 0.278652225
Statistical Computation B.7: The computation using a T-test assuming unequal variances
in terms of the rinsing capability of the oregano dishwashing liquid (Experimental Group)
against the commercial dishwashing liquid (Control Group)
Experimental Group Control Group

Mean 500 333.3333333
Variance 0 20833.33333
Observations 3 3
Hypothesized Mean 0
Difference
df 2
t Stat 2
P(T<=t) one-tail 0.09175171
P(T<=t) two-tail 0.183503419
APPENDIX “D”
Documentation
Figure 1. Collection of Oregano Leaves
Figure 2. Formulation of Oregano Dishwashing Liquid
Figure 3. Creating MacConkey and Mueller-Hinton Agar Plate

Figure 4. Putting Sensitivity Discs in MH Agar
Figure 5. Impregnation of Sensitivity Discs

Figure 6. Incubation of the MH Agar cultured with P. aeruginosa and S. areus
Figure 7. Results for Zone of Inhibition on P. aeruginosa and S. areus

Figure 6. Scrub Test Performance Figure 7. Foaming Capacity
Figure 8. Rinsing Capability

CURRICULUM VITAE
HERXIE YNA T. SUYAT

#272 Nalsian Centro, Calasiao, Pangasinan
09478800150
hy.suyat5@gmail.com
I. P E R S O N A L I N F O R M A T I O N:
Age : 18 years old
Birthdate : July 05, 2004
Sex : Female
Nationality : Filipino
Languages Spoken : Pangasinan, Filipino, and English
II. E D U C A T I O N A L B A C K G R O U N D:
Senior High School
Science, Technology, Engineering, and Mathematics
Malong St., Poblacion West, Calasiao, Pangasinan
2021 - 2023
Junior High School
Calasiao Comprehensive National High School (Special Science Program)
2017 - 2021
Elementary
Mother Lourdes Learning School
Nalsian Rd, Calasial, Calasiao, 2418 Pangasinan
2011 - 2017
CURRICULUM VITAE
BENEDICT S. PONTAWE
#578 Bued, Calasiao, Pangasinan
0992-247-9658
benedictpontawe1@gmail.com
<
Age : 17 years old
Sex : Male
Senior High School
2021 - 2023
Junior High School
2017 - 2021
Elementary
Bued East Elementary School
Bued, Calasiao Pangasinan
2011 - 2017
CURRICULUM VITAE
MHARIEL JEA P. CABALGADA
Address: Bued, Calasiao, Pangasinan
09305472701
cabalgadam@gmail.com
Age : 17 years old
Birthdate : September 29, 2005
Sex : Female
Senior High School
2021 - 2023
Junior High School
Calasiao Comprehensive National High School (Special Program in the Arts)
2018 - 2021
Bued National High School
299C+JHR, Brgy, Calasiao, Pangasinan
2017 - 2018
Elementary
Bued Elementary School
Bued, Calasiao, Pangasinan
2011 - 2017
CURRICULUM VITAE
Dhenielle Irish L. Gamboa

#190 Nalsian Centro, Calasiao, Pangasinan
09922038166
dhenielleirish@gmail.com
Age : 18 years old
Birthdate : March 17, 2005
Sex : Female
Senior High School
2021 - 2023
Junior High School
2017-2021
Elementary
Calasiao Central School
Pob East, Calasiao,Pangasinan
2011-2017
CURRICULUM VITAE
Adrian James A. Baldueza

#1414 Buenlag Calasiao Pangasinan
09212747369
adrianbaldueza2@gmail.com
Age : 17 years old
Sex : Male
Senior High School
2021 - 2023
Junior High School
Buenlag National High School
Buenlag Calasiao Pangasinan
2017 - 2021
Elementary
Buenlag Central School
2010 - 2017
CURRICULUM VITAE
CHRISTINE EDITH B. CANCINO

#413 Songkoy, Calasiao, Pangasinan
0154610100
kirstenncancino@gmail.com
Age : 17 years old
Birthdate : December 04, 2005
Sex : Female
Senior High School
2021- 2023
Junior High School
2017 - 2021
Elementary
Songkoy Elementary School
Songkoy, Calasiao, Pangasinan 2418 Pangasinan
2011 - 2017
CURRICULUM VITAE
HANS BENEDICT M. ANGUD
#161 Mancup, Calasiao, Pangasinan
0965-357-9463
hansangud@gmail.com
Age : 18 years old
Sex : Male
Senior High School
2021 - 2023
Junior High School
Calasiao Educational Center
Lasip Highway, Calasiao, Pangasinan
2017 - 2021
Elementary
Calasiao Educational Center
W.A. Jones, Nalsian, Calasiao, Pangasinan
2013 - 2017

Poblacion East, Calasiao Pangasinan
2011 – 2013
CURRICULUM VITAE
ANGEL SHAMER BARACE
#499 San Miguel, Calasiao, Pangasinan
0960-826-1608
angelshamerbarace@gmail.com
Age : 18 years old
Birthdate : January 15, 2005
Sex : Female
Senior High School
2021 - 2023
Junior High School
University of Luzon 2018-2021
Perez Blvd., Downtown Dist., Dagupan City, Pangasinan
Calasiao Comprehensive National High School 2017-2018
Elementary
Calasiao Central School 2015-2017
Caranglaan Elementary School 2011-2015
Caranglaan Dist., Dagupan City, Pangasinan

CURRICULUM VITAE
ERICKSON D. CABATBAT
#459 Buenlag, Calasiao, Pangasinan
09508391312
ericksoncabatbat@gmail.com
Age : 17 years old
Sex : Male
Senior High School
2022 - 2023
Junior High School
Elementary
2014 - 2017
CURRICULUM VITAE
GINALYN F. CABATBAT
#18 Talibaew, Calasiao, Pangasinan
0970-1127-638
ginalyn.ana.cabatbat@gmail.com
Age : 18 years old
Birthdate : February 28, 2005
Sex : Female
Senior High School
2021 - 2023
Junior High School
Comprehensive National High School (Special Science Program)

2017 - 2021
Elementary
2010-2017
CURRICULUM VITAE
PRINCESS A. BAROGO
969 Buenlag, Calasiao, Pangasinan
0939-508-3033
princessbarogo16@gmail.com
I. P E R S O N A L I N F O R M A T I O N
Age : 18 years old
Birthdate : August 14, 2004
Sex : Female
Nationality : Pangasinan, Filipino, and English
II. E D U C A T I O N A L B A C K G R O U N D
Senior High School
Science, Technology, Engineering and Mathematics
2021-2023
Junior High School
Buenlag, Calasiao, Pangasinan
2016-2021
Elementary
Buenlag, Calasiao, Pangasinan
2010-2016
CURRICULUM VITAE
CARLOS S. ARENAS
#737 San Miguel, Calasiao,Pangasinan
09052983761
imcarlosarenas.2005@gmail.com
I. PERSONAL INFORMATION
Age : 17 years old

Sex : Male
II. EDUCATIONALBACKGROUND
Senior High School
2021- 2023
Junior High School
2017 - 2021
Elementary
Pob.East, Calasiao,Pangasinan
2011-2017

As An Alternative Antibacterial Dishwashing Liquid - Suyat

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OREGANO (Coleus amboinicus) LEAVES EXTRACT AS AN ALTERNATIVE

ANTIBACTERIAL DISHWASHING LIQUID

A Research Proposal Presented to the Senior High School Department

In Partial Fulfillment of the Requirements for the Subjects

Herxie Yna T. Suyat

recommended for approval and acceptance for completion.

Approved on June 2, 2023, by the Panel for Oral Examination:

ENGR. MERIAN D. GALANG, PhD ENGR. JACKIELOUD D. DECENA

IVANSHANE L. MALALA GENESIS S. PAREL

CHRISTOPHER V. ZARATE HERNANDO C. ABALOS, JR.

DR. CHONA C. SAMSON, EdD

appreciation to the following people:

supplementary materials toward this project.

Andrei Dangoy and Brando Layda.

patient and significant pillars in conducting the study.

CHAPTER I: THE PROBLEM

A. Background of the Study……………………………………………………………….1

C. Statement of the Problem…………………………………………………………….

D. Hypotheses of the Study…………………………………………………………….

E. Scope and Delimitations…………………………………………………………………

F. Significance of the Study………………………………………………………………

CHAPTER II: REVIEW OF RELATED LITERATURE AND OTHER STUDIES

CHAPTER III: METHODOLOGY

C. Data Gathering Process……………………………………………………………….

D. Instruments for Data Collection………………………………………………………..

E. Statistical Treatment of data……………………………………………………………

A. Presentation and Analysis of Reports………………………………………………….

CHAPTER V: SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS

Appendix A – Communication Letters………………………………………………

Appendix C – Statistical Analysis……………………………………………………

Background of the Study

Conventional dishwashing liquids often contain synthetic chemicals and

surfactants that can be detrimental to aquatic ecosystems. According to the “Wastewater

has far-reaching consequences, including the disruption of aquatic life, contamination of

In addition, Zachariah (2020), claims many commercial dishwashing liquids

contain chemicals such as phosphates, formaldehyde, chlorine, and synthetic fragrances,

alternatives, including natural and nature-inspired antibacterial ingredients across their

kills germs with the advantages of aromaticity, safety, non-poison, high-efficiency

sterilization, and no compatibility contraindication. Oregano is an efficient antibacterial

ingredient in hand-washing and surface-cleaning detergent solutions and demonstrated

that it is a potential substitute for triclosan and chloroxylenol.

alternatives, there is a lack of comprehensive research exploring natural substitutes for

research focusing on their application as effective dishwashing agents. By addressing this

extract as an alternative antibacterial dishwashing liquid, providing a foundation for further

development and implementation of eco-friendly cleaning products.

oregano extract-based dishwashing liquid, which is expected to be biodegradable and

activity of oregano as a substitute in dishwashing liquids.

relation to the study, “Oregano (Coleus amboinicus) Leaves Extract as an Alternative

Antibacterial Dishwashing Liquid.”

Figure 1. Research paradigm of the Study, “Oregano (Coleus amboinicus) Leaves

The oregano (Coleus amboinicus) leaves extract acted as the independent

variable as it is composed of phytochemicals, carvacrol and thymol, that affect the

independent variable's effect.

This study aims to determine the effectiveness of oregano leaves extract as an

alternative antibacterial dishwashing liquid. Specifically, this study was conducted to

determine the following problems:

1. Is there any significant difference in the antibacterial activity of the treatment

groups, oregano leaves extract in dishwashing liquid, oregano extract, positive

Inhibition (ZOI) Test, against:

a. Staphylococcus aureus; and

b. Pseudomonas aeruginosa bacteria?

2. Is there a statistically significant difference between the dishwashing efficiency of

the oregano antibacterial dishwashing liquid to commercially available