Professional Documents
Culture Documents
CHAPTER 1
Introduction
therefore challenging local and international public health sector. The resistance of
pathogens to antibiotic remains unresolved problem in the world. The use of synthetic
drugs may subject the patient to a higher risk due to the unwanted toxicity. To address this,
actions must be taken, such as responsible use of antibiotics, developing new antibiotics,
and other polyphenolic compound that has an antimicrobial property. The substances
present in a plant can either inhibit the growth of bacteria or kill them, with minimum
toxicity to host cells. The metabolites from plant which exhibit a minimum toxicity are
This study aimed to formulate topical antibacterial ointment from the ethanolic
crude extract from the leaves of Piper betle Linn. locally known as Ikmo since there was a
study that Ikmo leaves have an antimicrobial property and shows no toxicity. Formulation
of a topical antibacterial ointment using ethanolic crude extract from the leaves of Ikmo
Skin infections are caused by a wide variety of germs, and symptoms can vary from
mild to serious. Mild infections may be treated with home remedies and/or an over-the-
counter preparations while other infections may require medical attention. There are
several types of skin infections, one of which is bacterial skin infections. Bacterial skin
infections often begin as small, red bumps that slowly increase in size. Some mild bacterial
infections are treated with topical antibiotics, but other infections are treated with oral
antibiotic. Common bacterial infections are cellulitis, boils, leprosy, and impetigo. (De
Impetigo is a highly contagious bacterial skin infection and is one of the most
common skin infections in children. Impetigo is usually caused by one of two bacteria:
Staphylococcus aureus or Group A streptococcus. There are several ways to prevent and/or
to treat impetigo. Antibiotics are the first line treatment when multiple lesions exist but due
to the high risk of antibiotic resistant bacteria, the management of impetigo in the future is
alternative treatment strategy, including the new topical antimicrobials. (Steele, 2017)
Ikmo leaves is a green, slender climbing plant, pungent in taste, and acrid in nature.
Africa, and Philippines. Ikmo leaves has been long used by the Filipino ancestors as an
important component of their “nga-nga”. In the Philippines, it is used together with lime
and betel nut to constitute the Filipino’s masticatory which helps in strengthening their
teeth and preventing cavities and halitosis. Nowadays, the piper betel is known for its many
3
medicinal uses like its antioxidant, antimalarial, cytotoxic, antifungal, antiseptic and
antibacterial property.
Conceptual Framework
This study aims to formulate an antibacterial ointment using Ethanolic crude extract
from Ikmo. Cases of antibiotic resistance in the world is continuously rising and resistance
of bacteria to antibiotic remains an unresolved problem. Skin infections can vary from mild
to serious. Mild infections may be treated with home remedies and/or an OTC preparations
while other infections may require medical attention. Antibiotics are usually the first line
treatment when it comes to skin infections but due to the high risk of antibiotic resistant
bacteria, the management of skin infections in the future is an area of concern therefore,
conducted the study, Formulation of topical antibacterial ointment using ethanolic crude
extract from the leaves of Ikmo with no previous studies of formulating ointment locally.
4
Piper betle
Ethanolic
Extract
Secondary Color
Staphylococcus Dermal Irritation
metabolites aureus Test
Appearance
Pseudomonas
aeruginosa
Solubility
Figure 1
Research Objectives
The main thrust of the study is to formulate an antibacterial ointment using ethanolic crude
Specific Objectives:
1. Obtain ethanolic crude extract form the leaves of Ikmo leaves and determine the
percentage yield;
2. Evaluate the ethanolic leaf extract using organoleptic, solubility and chemical test
(determination of metabolites);
4. Compare the antibacterial activity of the ethanolic crude extract against the
Hypothesis
crude Ikmo leaf extract and the standard drug mupirocin (p>0.05).
World Health Organization urges the public to search for new antimicrobial agents
therefore this study focuses on the formulation of topical antibacterial ointment using the
ethanolic crude extract of Ikmo leaves. Furthermore, the results of the study will give
To the Patients: Patient may use the formulated product derived from herbal plant
To the Pharmacists: Pharmacist can use this study to furthermore investigate and
produce new drug from natural source that has antimicrobial activities which is safe and
effective.
study to formulate antibacterial drugs derived from natural source that is cost effective than
and chemical test only. The ethanolic crude extract was undergone phytochemical
anthraquinone glycosides, and resins only. The researchers did not perform all the tests for
each screening. The researchers have only selected tests for each screenings.
The researchers used 20%, 40%, 60%, and 80% of the ethanolic crude extract. The
medium. The evaluation of ethanolic crude extract was done by measuring the diameter (in
millimeters) of zone of inhibition using Vernier caliper. The zone of inhibition of ethanolic
crude extract of Ikmo against Staphylococcus aureus and Pseudomonas aeruginosa was
interpreted based on the table provided from the manual (Source: Limuaco O.M, et.al.,
2014).
The researchers used the concentration that exhibited greater zone of inhibition in
formulation. The researchers used 60% ethanolic crude extract of Ikmo in formulation and
was compared to Mupirocin to see if there is a significant difference between two products.
The formulation was packed in collapsible tube and was properly labeled. The formulation
undergone dermal irritation test to evaluate its safety using rabbits as an animal model. All
six albino rabbits were examined for signs of erythema and oedema based on the grading
Definition of Terms
Antibacterial. Anything that kills bacteria or suppresses their growth or their ability to
reproduce. An agent that kills or suppresses the ability of bacteria to grow and reproduce.
Antibiotic resistance. The ability of bacteria and other microorganisms to resist the effects
Antioxidant. A chemical compound that protects cells against the effects of free radicals.
living tissue.
Boil. A pus-filled skin infection that starts in a hair follicle or oil gland.
Cellulitis. A spreading bacterial infection of the skin and tissues characterized by redness,
Impetigo. A common and highly contagious skin infection that mainly affects infants and
children.
Leprosy. A contagious disease caused by that affects the skin and peripheral nerves and
production of deformities.
Lesions. A region in an organ or tissue that has suffered damage through injury or disease,
Topical. A route of administration in which the site of drug intake is located beneath the
skin.
from plants.
10
Chapter 2
This chapter covers the compiled local and foreign literature and studies. This
chapter aims to present the information that the researchers were able to gather for better
In the past years, the use of medicinal plants, from different parts of the world, for
their antimicrobial property has been increasingly reported. According to the reports, the
plant extracts show a different target site other than those targeted by the synthetic
antibiotics and that these plant extracts targeting other sites with be effective against the
The modern era of antibiotics started with the discovery of Penicillin by Sir
Alexander Fleming in 1928. Since then, Antibiotics have transformed modern medicine
and saved millions of lives. Antibiotics were first prescribed to treat serious infections in
the 1940s. Antibiotics have not only saved patients’ lives, they have played a pivotal role
in achieving major advances in medicine and surgery. They have successfully prevented
or treated infections that can occur in patients who are receiving chemotherapy treatments;
who have chronic diseases such as diabetes, end-stage renal disease, or rheumatoid
arthritis; or who have had complex surgeries such as organ transplants, joint replacements,
endangering the efficacy of antibiotics, which have transformed medicine and saved
millions of lives. Many decades after the first patients were treated with antibiotics,
bacterial infections have again become a threat. The antibiotic resistance crisis has been
attributed to the overuse and misuse of these medications, as well as a lack of new drug
11
infections among World War II soldiers. However, shortly thereafter, Penicillin resistance
became a substantial clinical problem, so that, by the 1950s, many of the advances of the
prior decade were threatened. The overuse of antibiotics clearly drives the evolution of
antibiotic consumption and the emergence and dissemination of resistant bacteria strains.
In bacteria, genes can be inherited from relatives or can be acquired from nonrelatives on
mobile genetic elements such as plasmids. This horizontal gene transfer (HGT) can allow
(Ventola, 2015)
The infectious disease society of america has considered the following bacteria as
The same emerging MDR bacteria are also an immense threat in Asia. There have
culinary herbs from Thailand, Vietnam, and Malaysia. These culinary herbs are usually
consumed without proper heating. Acinetobacter baumannii, being able to exhibit a wide
12
spectrum of antimicrobial resistance mechanism, has also emerged as one of the most
problematic bacteria.
The current problem with the MDR bacteria is a serious, global medical crisis
which requires constant monitoring. Scientists are seeking for more natural or organic
materials as a solution for the diminishing efficacy and increasing toxicity of synthetic
drugs which further aggravate the problem. Traditional medicinal plants are being studied
further for the purpose of finding solution to the emerging problem regarding MDR
bacteria.
Bacteria was one of the most common cause of deadly diseases but with the
discovery of antibiotics, this problem was slowly resolved. However, due to inadequate
healthcare sector.
multiple class of drugs without considering remaining effective therapies. This leads to
bacterial strains being resistant to antibiotics. Some of the bacterial strains that are resistant
throat, intestine, vagina, and skin of human body. It is a round and bunched together
pathogen and has the ability to cause different kinds of mild to severe infections and it is
Pseudomonas aeruginosa is the most common pathogen isolated from patients who
have been hospitalized longer than 1 week, frequently causing nosocomial infections.
Some strains of MDR P. aeruginosa have been found to be resistant to nearly all
13
carbapenems.
Drugs intended for skin infections are usually formulated into ointments for topical
application. Ointments are greasy and they stay on the surface of the skin and are not well
absorbed, thus are “occlusive”. They are best used on dry skin and they trap moisture that’s
why they are able to keep the skin moist for longer periods of time. They have lesser chance
of causing allergy because of few preservatives. Ointments allow better penetration of the
active ingredient in the topical medication and they are better used on sensitive skin.
because skin and soft tissue infections such as scrapes and scratches or mild folliculitis, do
not usually require antibiotic treatment. In primary care, management should focus on good
skin hygiene, for patients with infected eczema, for wound management, or for other skin
infections, topical antibacterial medicines are not usually prescribed or they may only be
evergreen climbing vine whose height reaches 2 to 4 meters. Ikmo leaves, which has a
glossy heart-shaped physical appearance, is very popular in the Philippines where it is used
Ikmo is best grown in tropical areas like Philippines, Malaysia, Sri Lanka and
Indonesia. In the Philippines, it is best grown in region VI, in provinces like Ilo-Ilo,
Bacolod, and Cordillera Mountains. Its production is a very tedious process and it is not
The Ikmo leaf has been described to have piperol-a, piperol-b, methyl piper betlol.
The Ikmo leaves have starch, sugars, diastases and an essential oil composing of terpinen-
4-ol, safrole, allyl pyrocatechol monoacetate, eugenol, eugenyl acetate, hydroxyl chavicol
, eugenol , piper betol and the betle oil contains cadinene carvacrol, allyl catechol, chavicol,
Chavibetol None
Eugenol Yes
Hydroxychavicol Yes
Allypyrocathecol Yes
Quercetin Yes
Β-caryophyllene None
15
most important component of the essential oil from the leaves of the Ikmo plant. It is
an aromatic compound with a spicy odor and is an isomer of eugenol. (Dwivedi, Et. Al.,
2014)
Eugenol, one of the principal constituent of Ikmo leaf, has also been shown to
identified activities such as its anti-ulcerogenic potential and effect on osteoporosis and
especially its effect on the Central Nervous System (CNS) encompassing seizure control,
The new, immature leaves contains various beneficial bio- active compounds,
possesses anti carcinogenic, anti nitrosation, anti-mutagenic effects beside this, it has a
platelet and anti- thrombotic effects without impairing haemostatic function. In the aqueous
anticarcinogenic activities, whereas isolated from the chloroform withdrawal from aqueous
extract of Ikmo leaves show inhibitory action alongside oral cavity pathogens. 0.5%
pooled saliva the use of hydroxychavicol as an oral care agent. Hydroxychavicol show
compelling anti- inflammatory action by considerably inhibits the phrase of the pro
The phenolic constituent allyl pyrocatechol obtained from the leaves, show action
against obligate oral anaerobes responsible for halitosis. The leaf extract also has a
allyl pyrocatechol at different doses accelerates the rate of remedial of gastric lesion
flavonols. It occurs chiefly as glycosides, but other derivatives of quercetin have been
bioavailability of molecules when compare to the aglycone. Quercetin has also been
properties. The anticarcinogenic property of quercetin result from its important impact on
cancerous cell growth, decrease and alteration of cellular signal transduction pathways.
Animal evidence suggest quercetin’s antioxidant effects provides protection of the brain,
heart, and other tissues adjacent to ischemic a- reperfusion injury, toxic compounds, and
other factors that can persuade oxidative stress. Β-caryophyllene is a chief volatile
compound establish in huge amounts in different spice and food plants. (Dwivedi, Et. Al.,
2014)
with no genotoxic or cytotoxic effect in vivo. Clinical studies prove its efficiency in treating
selective and non- psychoactive full agonist for cb2 receptor in vivo. (Dwivedi, Et. Al.,
2014)
Through the discovery of antibiotics, millions of lives were saved but because of
the overuse and misuse of these medications, antibiotic resistance arose which is now,
again, endangering millions of lives. Multidrug resistance leads to strains of bacteria being
resistant to antibiotics. The current problem with the multidrug resistant bacteria is a
serious, medical crisis which requires constant monitoring. Some of the bacteria that are
Because of the emergence of antibiotic resistance, scientists are striving to discover a more
natural or organic source to be able to synthesize a drug that will solve the problem
associated with synthetically made drugs. In line with seeking for natural sources, various
studies about herbal plants are being conducted. One of the plants that have been used since
the early days is the piper betel. Piper betel, from the family Piperaceae, contains
antibacterial activity. Thus, piper betel may be used in formulating a medicine against
CHAPTER 3
This chapter is concerned with the research methodology, which covers the
topical antibacterial ointment from the ethanolic crude extract of Ikmo Leaves.
1. Determination of percentage yield ethanolic crude extract from the leaves of Ikmo.
The fresh Ikmo leaves were collected in the province of Nueva Ecija. The leaves
were washed with running tapped water, air- dried for 1 week, then grinded into powder
using blender. 500 g of Ikmo leaf powder in 2500 mL of Ethanol were mixed and macerated
for 48 hours. The mixture was subjected to filtration, then the filtrate was collected and
was incipient dried. The percentage yield was able to determine using the given formula
below.
Refer to Table1.
38.0 grams of the medium were suspended in 1000 mL of distilled water. Heat to
boiling to dissolve the medium completely. The medium was sterilized using an autoclave
at 121°C for 15 minutes, then cooled to 45 to 50°C and poured into sterile petri plates.
Staphylococcus aureus and Pseudomonas aeruginosa are bacteria used in the study.
The inocula used were undergone confirmatory test using agar plates to determine its
purity. Nutrient agar was used to test the purity of Pseudomonas aeruginosa, and on the
other hand, Mannitol Salt Agar was used for Staphylococcus aureus. The researchers
so that the number of bacteria will be within a given range to standardize microbial testing.
3.2.1 Inoculation
The researchers used plated medium technique; made a zigzag streak on the surface
Mupirocin was used as a reference standard because Mupirocin was the most
Different concentrations of ethanolic crude extract of Ikmo. (20%, 40%, 60%, 80%)
and, the water and tween 80 as the negative control were placed on the agar plate
individually and covered aseptically. The plates were incubated at 35-37°C for 16-18
hours. The antibacterial activity was determined by measuring the diameter of the zone of
20
inhibition in millimeters using a vernier caliper. The result was interpreted based on the
table provided.
TABLE 2
Diameter of Zone of
Activity Interpretation
Inhibition (mm)
< 10 Inactive
14 – 19 Active Intermediate
The study aims to formulate and evaluate an antibacterial topical ointment based
on the following:
The extract was incorporated into the molten simple ointment base and allowed to
Formulation of Ingredients
Modification in the Formulation of the White Ointment using the Ethanolic Crude
Extract of Ikmo
COMPOUNDING PROCEDURES
4. Incorporate the extract into the molten simple ointment base and stir the mixture until it
begins to congeal.
The researchers used six (6) male albino rabbits to test the formulated ointment
with the active ingredient and the formulation containing the excipients only to be
conducted in three (3) trials. The rabbits were acclimatized for seven (7) days. Twenty four
(24) hours prior to testing, their backs was shaved and a pinch of the formulation was
applied on the shaved area and covered with gauzed patch. The animals should be observed
up to 14 days after removal of the patches. The results were examined for signs of erythema
and oedema based on the grading of skin reactions provided from OECD/OCD and the
responses scored at 60 minutes, and then at 24, 48 and 72 hours after patch removal.
22
5. Statistical Analysis
the Ikmo ethanolic crude extract and Mupirocin as the positive control.
the average zones of inhibition of the Ikmo ethanolic crude extract and the positive control,
Mupirocin.
23
Piper betle
Ethanolic
Crude
Extract
Figure 1
Chapter 4
This chapter provides the presentation, analysis and interpretation of the data
gathered from the tests conducted to the ethanolic crude extract from the leaves of Piper
betle Linn.
1. Percentage Yield of Ethanolic Crude extract from the leaves of Piper betle Linn.
The Ethanolic Crude Extract from Piper betle Linn. were obtained by maceration
using 95% ethanol as solvent then was evaporated to incipient dryness. The ethanolic
crude extract was weighed and the percentage yield was computed. The percentage
TABLE 1
(Evdokimor, 2014)
The ethanolic crude extract was soluble in acetone, ethanol, and ether and insoluble
TABLE 2
in water.
substance
constituents
organic metabolites
26
TABLE 3
Brick-red
Formation of brick red Presence of
Fehling’s Test precipitate precipitate Carbohydrates,
reducing
sugars
27
Absence of
Xanthoproteic Formation of No Formation of yellow proteins,
color specifically
Test yellow precipitate aromatic
amino acids
For the primary metabolites, the ethanolic crude extract of Ikmo leaves contains
carbohydrates, specifically reducing sugars, while it can be noted that proteins are not
detected primarily due to the application of heat during the preparation of crude extract
precipitate
A formation of
Phloroglucinol
reddish-brown Reddish-brown color Presence of Resins
Test
color
The ethanolic Ikmo leaf extract contains polyphenols like flavonoids, tannins and
3. Determination of the antibacterial property of the ethanolic crude extract from the
Four different concentrations (20%, 40%, 60%, and 80%) of ethanolic crude extract
of Ikmo have the capability of inhibiting both Staphylococcus aureus and Pseudomonas
aeruginosa.
aureus.
aeruginosa.
31
4. Statistical Analysis: Comparison between the Ethanolic Crude Extract and the
Positive Control
Staphylococcus aureus
Pseudomonas aeruginosa
The table shows the increasing dose of the ethanolic crude extract from the leaves
of Ikmo against S. aureus and P. aeruginosa. The results showed that all concentration of
Ikmo were effective against S. aureus and P. aeruginosa and statistically comparable to
the standard drug, mupirocin. The P-value of 20% extract (1.26E-04 and 4.52E-06), 40%
extract (5.45E-05 and 7.97E-08), 60% extract (2.21E-05 and 1.13E-08), 80% extract
respectively showed significant difference against the positive control. Moreover, the
researchers used the 60% concentration in preparing the antibacterial ointment since it has
The formulation was prepared by incorporation method. 60% of the ethanolic crude
extract of Ikmo was used as an active ingredient and white ointment was used as an
ointment base. The extract, white wax, and white petrolatum were mixed together and
allowed to congeal by stirring. The formulation was packed in collapsible tube and labeled
The formulation was undergone antibacterial susceptibility test and was compared
to mupirocin ointment. The 60% Ikmo ointment has the capability of inhibiting
(17.55±0.03mm).
chemical was removed, using water. The rabbits were expected to observe up to 14 days
after removal of the patches. The experiment was terminated in the 7th day since the hair
skin of the rabbit started to grow. All six albino rabbits were examined for signs of
erythema and oedema according to the grading of skin reactions from OECD, and the
responses scored at 60 minutes, and then at 24, 48 and 72 hours after patch removal.
Test chemical:
60% Ikmo Leaf Extract Ointment
Vehicle:
White Petrolatum
Test animals:
(6) Male albino rabbits from Department of Science and Technology (DOST) with
weight of NLT 1.5 kg were used.
Test condition:
The patch was loosely held in contact with the skin by means of a suitable semi-occlusive
dressing. Access by the animal to the patch and ingestion or inhalation of the test
chemical was prevented.
Results: 24 h 48 h 72 h
C
Normal Skin Applied with
Simple ointment
base
Rabbits A, B, and C were examined for signs of erythema and oedema and graded
according to the grading of skin reactions from OECD. Rabbits A, B, and C have seen no
erythema and oedema formation.
34
CHAPTER 5
This chapter contains the summary of findings from the performed tests, the
Summary of Findings
Perfectly mature Piper betle, L. leaves were collected, dried and macerated using
95% ethanol and subjected to reflux distillation. The extractives were dried to incipient
dryness and the percentage yield of the extract was 9.9222%. The Ikmo leaves extract is
greenish-black, with distinct creosote-like odor and has syrupy consistency. The ethanolic
crude extract of Piper betle Linn. Leaves (Ikmo) contains reducing sugars, flavonoids,
tannins, alkaloids, and resins. The microbiological assay was conducted using agar-plate
method using two (2) test organisms, namely S. aureus and P. aeruginosa. The assay was
Increasing dose of the ethanolic crude extract (increments of 20%) of Ikmo were
used and exhibited an average zone of inhibition of 25.85±1.13 & 20.63±0.58 mm,
26.82±1.56 & 21.98±0.55 mm, 27.92±1.36 & 22.83±0.47 mm and 25.38±3.91 &
The 60% concentration was used in formulation an ointment and was tested on rabbits. The
formulated ointment did not show erythema and oedema formation on the skin of the
rabbits.
35
Conclusion
Based on all available variables in this study, the ethanolic crude extract of Ikmo
mupirocin. Furthermore the formulated ointment did not show any irritation to the
Recommendations
For the further improvement of the presented results of this study, the researchers
thereby recommend the following: (1) improvement of the appearance of the formulated
ointment; (2) test the antibacterial activity of the formulation against Streptococcus
pyogenes and MRSA Staphylococcus aureus; (3) conduct in vitro-release testing for the
formulated antibacterial ointment, (4) conduct stability testing for the formulated
antibacterial ointment and (5) conduct stability test of the excipients and active drug.
36
REFERENCES
37
Fawad, et al., (2012) In-Vitro Antibacterial Activity of Piper betle Leaf Extracts.
Journal of Applied Pharmacy. p. 643
Valle, et al., (2016) Antimicrobial Activities of Methanol, Ethanol, and Supercritical CO2
Extracts of Philippine Piper betle L. on Critical Isolates of Gram Positive and
Gram Negative Bacteria with Transferable Multiple Drug Resistance. p. 1.
Nalina, T., et al., (2007) The Crude Aqueous Extract of Piper betle L. and its Antibacterial
Effect Towards Streptococcus mutans. pp. 10-11.
Budiman, A., et al., (2018) Antibacterial Activity of Piper betle L. Extract in Cream
Dosage Forms against Staphylococcus aureus and Propionibacterium
acne. p.
Aryal, S., (2018) Mueller Hinton Agar (MHA) – Composition, Principle, Uses, and
Preparation.
Pawar, S., et al., (2017) Biochemical Profiling of Antifungal Activity of Betel Leaf (Piper
betle L.) Extract and Its Significance in Traditional Medicine. Journal of
Advanced Research in Biotechnology. p. 1
Kaveti, B., et al., (2011) Antibacterial Activity of Piper betle Leaves. International Journal
of Pharmacy Teaching and Practices. p. 129
Blas, E., et al., (2016) Antifungal Property of Polyphenolic Compo unds from Ikmo (Piper
betle fam. Piperaceae) Leaves and Formulation of Cream. pp. 1, 10, 11, 12
Limuaco O.M, et.al., (2012) Laboratory Exercises in Pharmacognosy and Plant Chemistry.
pp. 23, 25, 26, 27, 28, 29
Dwivedi V, et. Al., (2014) Journal of Pharmacognosy and Phytochemistry
pp. 93, 94, 94
38
APPENDICES
39
APPENDIX A
Plant Sample
Ikmo
Family: Piperaceae
APPENDIX B
APPENDIX C
41
Certificate of Authentication
42
APPENDIX D
IACUC PROTOCOL
43
APPENDIX E
Maceration
Filtration
Evaporation
44
APPENDIX F
Solubility
Water Acetone
Ethanol Ether
45
APPENDIX G
Phytochemical Screening
APPENDIX H
Microbiological Screening
Staphylococcus aureus
Negative Control
Positive Control
20%
54
40%
60%
80%
55
Pseudomonas aeruginosa
Negative Control
Positive Control
20%
56
40%
60%
80%
57
APPENDIX I
Drug
Staphylococcus aureus
Negative Control
Positive Control
Pseudomonas aeruginosa
Negative Control
Positive Control
APPENDIX J
Statistical Treatment
60
61
62
APPENDIX K
Formulation of Ingredients
Modification in the Formulation of the White Ointment using the Ethanolic Crude
Extract of Ikmo
APPENDIX L
Day 1
Positive
Negative
111
Day 2
Positive
64
Negative
Day 3
Positive
Negative
Negative
65
Day 4
Positive
Negative
Day 5
Positive
66
Negative
Day 6
Positive
Negative
67
Day 7
Positive
Negative
68
APPENDIX M
Plagiarism Scan
SOURCE: https://www.plagscan.com/docman
Content Percentage
Chapter 1
The Problem and Its 1846 words 9.1 %
Setting
Chapter 2
Review of Related 1720 words 8.5 %
Literature and Studies
Chapter 3
1715 words 8.8 %
Methods and Procedures