St.

Louise de Marillac College of Bogo,

Cebu, Inc.
Sor Dorotea Rubio Street, Bogo City, 6010, Cebu, Philippines
Contact Numbers: (032) 434-7160, (032) 260-1365 or 0967-012-7261
Member: Daughters of Charity – St. Louise de Marillac Educational System (DC-
SLMES)

Utilization of Premna odorata as an Organic Alternative to

Synthetic Chemical Disinfectants

In Partial Fulfillment of the Requirements in Research Capstone

for Science, Technology, Engineering, and Mathematics Strand

Balaga, Darcy Marie

Barte, Angelique

Isnani, Enar Josef

Lepon, Daphne

Lepon, John Ernest

Mangubat, Nicole Franscine

Pogado, Shaina Marie

Varga, Dinzyl Babe

April 2022

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 CHAPTER I

INTRODUCTION

In today’s world, medicine is considered as one of the dependent ways to

cure a person of illness. It is evident that the majority of old and poor people still
refer to traditional herbal medicine as an alternative, due to not affording or
trusting modern medicine and its effectiveness. But scientists are researching
medicines that are effective at the same time cheap so that anybody can benefit
from accurate cure efficiency. So, they are researching plants that produce
antibodies.

Antibodies are an important component of vertebrate's adaptive immune

systems, and they may now be made by converting plants with antibody-coding
genes from animals and humans. Despite the fact that plants do not produce
antibodies naturally, plant-derived antibodies (plantibodies) have been proven to
behave similarly to mammalian antibodies.

An example of a plant that has plantibodies is Premna odorata. P. odorata

Blanco (Verbenaceae) is a native plant of temperate and tropical Asia which
includes the Philippines. P. odorata, is primarily used in folk medicine to treat
inflammation, wound healing, and skin diseases. Many in vitro and in vivo
investigations have been carried out to determine the biological and
pharmacological characteristics of Premna species extracts and identified
compounds with antibacterial, antioxidant, anti-inflammatory,
immuno-modulatory, anti-hyperglycemia, and cytotoxic activity. This chapter
includes the dependent and independent variables that are used in the study as
well as the significance and limitations. It presents the introduction of the
research study and its uses.

In this study, we shall conduct the utilization of Premna odorata as a

disinfectant and research if it can be made as an alternative disinfectant that can
reduce microorganism that cause diseases.

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A. Background of the study

CoViD-19 has been ongoing for the past year and people are hoarding
disinfectants for their own safety. As time goes on disinfectants stocks will
eventually go down and it is considered as one of the defense mechanisms
against germs, bacteria and fungi. To fight this horrible scenario, we can convert
into antibody-making plants and study if they are reliable as disinfectants.

The World Health Organization's (WHO) estimate indicates that 80% of

more than 400 million inhabitants of the world rely chiefly on traditional medicines
for their primary health care needs (WHO, 2008). These statistics prove that
people rely on natural antimicrobial rather than lab created antibiotics. The
change of modalities can surely encourage people who trust traditional medicine
to use the created disinfectant. Western medicine is increasingly being receptive
to the use of antimicrobials and other drugs derived from plants, as mainstream
antibiotics are becoming ineffective due to the emergence of resistant strains.
The developments of resistant strains have made diseases intractable to
conventional antimicrobials (Bansal et al., 2012).

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B. Statement of the Problem

This study was aimed to determine the abundance and its underlying
parameter of Premna odorata as a disinfectant. Specifically, it aimed to answer
the following:

1. What are the properties of P. odorata that can prove to progress into an
alternative disinfectant?

2. How can we use P. odorata as a disinfectant?

3. Is P. odorata capable of being an organic disinfectant, considering its

antiseptic properties?

4. Is there a significant change in using P. odorata as an alternative

disinfectant if proven to be capable?

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C. Research Objectives

This study aims to determine and answer the following:

To identify the acceptability of Premna odorata as an additive-free

disinfectant that can be a substitute or an alternative for chemical disinfectants
such as bleach, chlorine, vinegar, or hydrogen peroxide.

Specific objectives:

a. To identify the properties of P. odorata that are able to reduce the number of
bacteria, viruses, or fungi to a desired concentration, considering the midst of the
pandemic. 

b. To recognize how this organic disinfectant helps protect the environment,

safeguard the health of consumers or residence, and improve indoor air quality.

c. To describe how P. odorata can affect the competitiveness in pricing as the

demand will decrease (if proven perpetrate as a coequal disinfectant identical to
bleach, detergent, hot water, etc.)

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D. Statement of Hypothesis

Na: There is a significant result on Premna Odorata as a disinfectant and if it can

be made as an alternative disinfectant.

No: There is no significant result on Premna Odorata as a disinfectant and if it

can be made as an alternative disinfectant.

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E. Significance of the Study

The research is made with aim to provide crucial information and

knowledge regarding the chosen topic from the researches recent studies or
theses , and related sites needed for the expected importance to the following:

As a disinfectant, this study seeks to know the properties of Premna

odorata if it can be used and can be capable of being a disinfectant or not.

To the people, this study will help the public to be aware that we can have
this idea and save a lot of money from buying disinfectants like alcohol,
especially knowing that disinfectants are highly in demand nowadays.

The researchers and future researchers, the findings of this study will help
researchers determine on improving the study by experimenting on Premna
odorata plants. It intends to help the department of medicine be aware of
everyone’s perspective and uphold new things to improve their medicine. This
study would also help the researchers' queries and suggestions about the P.
odorata as an additive-free disinfectant.

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F. Scope and Delimitation

This study is mainly focused on testing if components found in Premna

odorata can be used as an alternative, organic all-around disinfectant and if so,
how we can make it into an all-around disinfectant.

It makes use of the different parts of the P. odorata, from the shoots and
leaves, to the bark and roots of the plant. It is necessary to utilize and test all of
the parts so that we can test on what part has the most potential for extraction of
the needed components to create the disinfectant substitute.

The processes included the testing of the raw decoctions coming from the
different parts of the plants and testing it on various pathogens such as bacteria,
viruses, parasites, fungi, protozoa, and pests.

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G. Definition of Terms
The following terms were defined according to how they are being used in the
study:

Anti-hyperglycemia - tending to lower elevated blood glucose levels.

Antioxidant - substance that protects cells from the damage caused by free
radicals (unstable molecules made by the process of oxidation during normal
metabolism).
Chemical – a compound or substance that has been purified or prepared
artificially.
Chemical reaction - is a process in which one or more substances, also called
reactants, are converted to one or more different substances, known as products.
Cytotoxic - refers to a substance or process which results in cell damage or cell
death.
Disinfectant – a liquid that destroys pathogens.
Immuno-modulatory - change in the body's immune system, caused by agents
that activate or suppress its function.
Organic – deriving from living matter is naturally occurring.
Parameter - a numerical or other measurable factor forming one of a set that
defines a system or sets the conditions of its operation.
Pathogens – an organism that causes disease.
Synthetic – of a substance made by chemical synthesis, especially to imitate a
natural product.
Underlying - significant as a cause or basis of something but not necessarily
manifest or obvious.
Vertebrate - any animal of the chordate subphylum Vertebrata, which includes
the fishes, amphibians, reptiles, birds, and mammals.

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 CHAPTER II
REVIEW OF RELATED LITERATURE

A. Related Literature

The study of Premna species and their antibodies has good effectiveness
against certain bacteria and viruses. Its effectiveness in becoming a disinfectant
is tested when the antiparasitic activity of some new Caledonian medicinal
plants including Premna serratifolia were evaluated. It was observed that P.
serratifolia was active against Leishmania donovani with IC50 values between
0.5-5µg/ml (Desrivit et al., 2007).

In researching on alternative disinfectants, they discovered the

antibacterial activity of hexane, chloroform, ethyl alcohol, and water extracts from
the stem bark of Premna tomentosa was done and it was reported that
chloroform extract of the root inhibited the growth of Enterobacter aerogenes but
there was no activity on the other extracts. The presence of di-C-glycosyl
flavones in the heartwood of the plant might be the reason for its antibacterial
activity. Also, all the extracts were active against Alcaligenes faecalis, Bacillus
subtilis and Escherichia coli (Anbazhakan and Bahu, 2007).

The abundant use of anti-infective agents resulted in the emergence of

drug-resistant bacteria, fungi, and viruses. To overcome the increasing resistance
of pathogenic microbes, a variety of medicinal plants have been screened
worldwide for their antimicrobial properties. The aim is to find new, effective
antimicrobial agents with novel modes of action. Essential oils derived from
aromatic medicinal plants have been reported to exhibit exceptionally good
antimicrobial effects against bacteria, yeasts, filamentous fungi, and viruses. The
progress of this expanding scientific field will be recorded by the most important
results published in the last decade. (S. Karger AG, Basel, 2009).

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B. Related Studies

Local Study
According to Erna et al. (2017) selected Philippine plant extracts as
alternative preservatives for a pharmaceutical liquid preparation. Philippine
Journal of Science 146 (1) Preservatives play an essential role in enhancing
quality and prolonging shelf-life of pharmaceutical products by improving their
antimicrobial stability or reducing the amounts of oxidative degradation products.
Persistent use of synthetic compounds as preservatives resulted in several
reports of undesirable effects. Hence, development of alternatives is necessary
to maintain their vital function while minimizing adverse effects. In this study,
ethanolic extracts of five plants with known antimicrobial activities, Psidium
guajava, Premna odorata, Mimosa pudica, Allium sativum, and Zingiber
officinale, were formulated into suspensions and evaluated for preservative
activity using the United States Pharmacopeia (USP)(2015) guidelines.
Phytochemical test, antioxidant activity and compatibility test were also
conducted on the extracts. P. odorata (p= 0.999) and M. pudica (p= 0.054) at
5.00 mg/mL concentration exhibited comparable antioxidant activity against the
standard antioxidant preservative, butylated hydroxytoluene, using ferric
reduction antioxidant power assay. Based on the criteria for product category 4 of
the USP, suspensions of P. odorata and P. guajava demonstrated acceptable
preservative activity against selected microorganisms, Escherichia coli and
Staphylococcus aureus. These bioactivities can be attributed to the
phytochemicals present in the extracts such as glycosides, reducing substances,
flavonoids and alkaloids. In conclusion, for the USP category 4 products such as
antacid suspensions, P. guajava can be utilized as an alternative source of
antimicrobial preservative, M. pudica as an alternative source of antioxidant
preservative, and P. odorata as an alternative source of preservative with both
antimicrobial and antioxidant efficacy.

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Foreign Study

Premna odorata is one of the seven components of a commercialized

Philippine herbal preparation called “Pito-Pito”. The ethnomedicinal uses of P.
odorata, however, have not been scientifically validated. Apart from its
uncommonly known ethnomedicinal uses, this tree does not have much
economic value and is generally cut and replaced with plants that are
perceived to be more profitable. For this reason, even though it is a native
plant, it is not generally cultivated and not commonly found in populated
areas, but thrive in the less accessible secondary limestone forests of the
Philippines. The isolation of bioactive and medicinal compounds from the
leaves of P. odorata would provide scientific evidence of the medicinal
properties of the plant. The validated medicinal properties, in turn, could
become the popular motivation for the plant’s sustainable use, conservation
and cultivation. The current study was able to isolate two flavone
aglycones, diosmetin and acacetin, from the crude EtOH extract of the leaves.
Diosmetin or 5,7,3’-trihydroxy-4’-methoxyflavone was classified as a
non-widespread flavone (Valant-Vetschera and Wollenweber, 2006). Many
earlier studies have reported its bioactivities. It inhibited 3H-dopamine uptake
in control and differentiated neuroblastoma cells and in small-cell lung
carcinoma cells. It also inhibited 3H-serotonin uptake in both cell types.
These inhibitory effects could be responsible for the increased vascular tone
observed in vivo after treatment with diosmetin and its glycoside, diosmin, as
vasotonic agents (Sher et al., 1992). As an antimicrobial agent, diosmetin
exhibited the MIC at 25 µg/mL against B. subtilis and the MIC at 50 µg/mL
against the fungus Trichophyton rubrum, the most common cause of athlete’s
foot (Meng et al., 2000). It down-regulates the enzyme cyclooxygenase-2
and so could be an anti-inflammatory agent (Lopez-Posadas et al., 2008). It
increased osteoblast differentiation and so could be a potential agent for
treating osteoporosis (Hsu and Kuo, 2008). At 60 µM, it exhibited 55%
inhibition of the enzyme α-phosphatidylinositol-3-kinase (Agullo et al., 1997).

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At 6 µM, it exhibited 70% inhibition of the enzyme 17-β hydroxysteroid
dehydrogenase type 1. As a chemo preventive compound, diosmetin directly
inhibited the activity of the enzyme cytochrome P450 1A1 (Ciolino et al., 1998).
It blocked apoptosis that was induced by dimethylbenzene (a)anthracene
(Ciolino et al., 2002). It exerted cytostatic effects on cell cycle progression
and proliferation of breast cancer cells (Androutsopoulos et al., 2009).
Acacetin or 5,7-dihydroxy-4’-methoxy-flavone was classified as a widespread
flavone (Valant-Vetschera and Wollenweber, 2006). Several bioactivities of
acacetin have been reported by earlier studies. As an antifungal agent,
acacetin had a comparable potency with the antifungal drug Amphotericin B
against Candida glabrata KCTC 7219, Candida tropicalis KCTC 7725 and C.
tropicalis KCTC 7212 (Rahman and Monn, 2007). It was reported to be a
promising agent for the treatment of atrial fibrillation (Li et al., 2008). As a
chemopreventive agent, acacetin had antiproliferative effect on human liver
cancer cell line HepG2 (Hsu et al., 2004a) and in human non-small cell
lung cancer A549 cells (Hsu et al., 2004b). It was capable of preventing
inflammation-associated tumorigenesis (Pan et al., 2006).

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 CHAPTER III
METHODOLOGY

,
A. Research Design

The researchers have utilized the experimental research design for this
study. The researchers manipulate the independent variable that is the different
product formulations of the Premna odorata. Fain, A., (2010), stated that
experimental research is a study that strictly adheres to a scientific research
design. It includes a hypothesis, a variable that can be manipulated by the
researcher, and variables that can be measured, calculated and compared. Most
importantly, experimental research is completed in a controlled environment. The
researcher collects data and results will either support or reject the hypothesis.

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B. Subjects of the Study

In this experimental study, four (4) petri dishes with different pathogens,
being the subjects of the study, are placed in a sterile and controlled environment
with 1 clean dish as the control variable. The subjects will be then sprayed with
the created disinfectant.

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C. Sampling Technique

The sample size will be determined using the Slovin's Formula in the 50%
(e=0.50) confidence interval.

Sample Size Formula:

N=

Where:

n = sample size

N = total population value

e = confidence interval

I = constant value

The sample size of the study where the total population (N) of different
types of pathogens contained in separate Petri dishes will be determined and
confidence interval is at 0.50 is 5 percent of pathogens that will build up in the
solution.

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D. Research Instrument to be Used

The researchers prepared a few experiments to test the bacteria cultured

by the researchers in terms of Premna odorata as a Disinfectant.

The researchers did this to determine the effectiveness of the Utilization of

P. odorata as a Disinfectant to the cultured bacteria.

The researchers innovatively gathered data from the experiment wherein it

clarifies the potential of P. odorata as a disinfectant. Since the instrument crafted
was self-made it is subjected for validation.

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E. Procedure of Data Gathering
The researchers allotted their time, efforts, and cooperation in developing
cultured bacteria with explicit purpose so as to serve its intended role in the
experiment. With restrictions due to health protocols, the researchers opted for
making their own instruments. The researchers first determined which part of P.
odorata has the most potential for extraction needed to create a disinfectant.
Three (3) petri dishes were cultured using samples gathered from a kitchen
countertop. The parts that were tested are the new growth and leaves, bark, and
roots, the procedure of extraction was to simmer the parts for an hour. The
researchers discovered that the new growth and leaves have the most potential.
Then the researchers utilized eight (8) petri dishes, four (4) treated, and four (4)
not treated with different pathogens, they were placed in a sterile and controlled
environment. The researchers then simmered 50 grams of P. odorata leaves and
500mL water, as the extract that will be considered as the disinfectant. The 4
petri dishes with different pathogens were then sprayed with the created
disinfectant. Afterwards, the researchers left the treated plate in the controlled
environment, and after a week, and found significant changes. The instruments
were retrieved after they were finished experimenting.

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F. Statistical Treatment

Data from the experiment by the researchers were statistically analyzed

with the requirements of the study. Researchers statistically analyzed the data
instruments of the study to treat the data gathered from the experiment of the
Utilization of Premna odorata as a Disinfectant. Through this treatment, the
researchers were able to determine the effectiveness and if there is a correlation
between the independent and dependent variables of Utilization of P. odorata as
a disinfectant to the people as the information was tabulated and statistically
analyzed.

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G. Collection of Materials

The researchers gathered the data and information from previous studies
and observation of other researchers who seized the medicinal potentiality of
Premna odorata. Furthermore, the researchers also convene data from articles,
statistical reports regarding the native trees of the Philippines, and dictionaries
from certain terms used in the content.

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H. Preparation of Treatment

The treatment that was prepared in this study is needed for data collection
that could capture the quality evidence that seeks out variables, and viability of
the data. The collected information and folk knowledge regarding the medicinal
and cleansing uses of the indigenous plant, Premna odorata, can aid in the
reduction of the number of bacteria, viruses, or fungi to a lesser concentration.
The scientific evidence of the cleansing properties of the leaves of P. odorata
substantiated by the study, can be handed down from generation to generation
through verbal/virtual communication, its impact on how its organic disinfectant
help protect the environment, safeguard the health of consumers or residence,
and improve indoor air quality. The researchers collected a methodical process
of gathering and analyzing the specific information of the study to proffer
solutions and evaluate the results of P. odorata as an organic additive free
disinfectant.

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 CHAPTER IV

PRESENTATION, ANALYSIS AND INTERPRETATION OF DATA

This chapter is concerned with the data presentation of the findings

obtained through the study, and therefore contains the data gathered from the
experiments conducted that would be beneficial to our study. The experiment
trialed on three types of decoctions containing components from the various
parts of the Premna odorata, namely: the leaves and new growth, the bark, and
the roots. Researchers then tested with the use of cultured bacteria plates along
with the chosen decoction that produced the most promising results, which was
the combination of leaves and new growths, proving it’s effectiveness of being a
natural disinfectant against bacteria and other pathogens.

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 The bar graph shows the growth rate and size of the cultures grown,
wherein the cultured agar plates were applied a decoction made from different
parts of the Premna odorata, to test which parts of the plant are the most viable
and has the most potential for the creation of a natural disinfectant. The agar
plate applied with the decoction created with the new growth and leaves of the
Premna odorata, showed the least amount of growth, with the culture only filling
20% of the agar surface. The cultured agar plates applied with the decoction
created with the bark, and the decoction created with the leaves of the Premna
odorata, showed more growth compared to the one applied with the decoction
created with the new growth and leaves, with the culture filling 80% and 60% of
the agar surface, respectively.

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 The bar graph shows the growth rate and size of the cultures grown,
wherein samples were gathered from various high-touch locations. There was a
significant difference between the treated and untreated plates, with the former
showing less growth and the latter having more growth. The plates with Swab A
showed the most growth, with the untreated plate having filled 90% of the total
growing surface of the plate, and the treated plate having filled 40%. The plates
with Swab B showed less growth compared to the plates with Swab A, with the
untreated plate having filled 60% of the total growing surface of the plate, and the
treated plate having filled 20%. The plates with Swab C showed the least growth,
with the untreated plate having filled 40% of the total growing surface of the
plate, and the treated plate having filled 10%. Lastly, the plates with Swab D, with
the untreated plate having filled 70% of the total growing surface of the plate, and
the treated plate having filled 30%.

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