Antibacterial Effect of Euphorbia hirta (Tawa Tawa) Leaves Extract

on Minor Open Wounds

A. Science Topic Proposal

(LIFE SCIENCE)

Presented to:

MRS. LORRAINE A. VOLOSO

Research Adviser

Presented by:

XANDREA ROMEEN G. SALINAS

Researcher
Grade 9-Atis

School Year 2021-2022

GOVERNOR FERRER MEMORIAL NATIONAL HIGHSCHOOL – MAIN

Pinagtipunan, City of General Trias Cavite
 ABSTRACT

The purpose of the study is to help people who are experiencing open wound accidents. The
main variable of this study is Tawa-Tawa (Euphorbia Hirta) leaves extract, which is known and
used as an agent to several pharmacological properties, specifically, Anti-Bacterial agent.
Euphorbia Hirta is a type of herbal plant that may be abundant in growing to any places
because of the climate in the community. The researcher first washed the Tawa-Tawa leaves.
About 20 grams of these were pounded using the mortar and pestle. Then, the pounded leaves
were boiled for 30 minutes with 120 oz of water. The researcher extracted the juice from the
leaves by filtration using a clean cloth and a funnel. After filtrating, the researcher divided the
juices with different concentrations into three (3) containers to be used in the three (3) trials.

The first group (Trial 1) was weighed at about 1 ml of tawa-tawa leaves extract with 9 ml
of water.
Second group (Trial 2) was weighed at about 2 ml of Tawa-Tawa leaves extract with 8 ml
of water.
Third group (Trial 3) was weighed at about 3 ml of Tawa-Tawa leaves extract with 7 ml of
water.

In conducting Test and Trial, the researcher asked for assistance in cutting an open wound on
the three (3) laboratory albino mice.
After creating the wound, the researcher applied the three (3) TawaTawa wound treatment to
the three (3) laboratory albino mice. After the 1st part of test and trials, the researcher observed
the data presented from the wound of mice. The Test and Trials on the three (3) Laboratory
mice was conducted and observed for three (3) days.

The researcher applied each with Tawa-Tawa wound treatment every day. The researcher
critiqued the data collected based on the effectiveness, time, skin reactions, and healing
stages of an open wound. After the results, the researcher used the concentration of Tawa-
Tawa treatment used in Trial 3 to create a comparison between the created product and the
commercial product. The researcher conducted the final trial (comparison of products) to the
two (2) laboratory albino mice. The researcher observed the healing reactions from the
experimentation.

The possible positive outcome of this experiment may be deemed beneficial to the community,
especially to people who may not be able to afford commercial wound treatment products. The
self-help application of this herbal product may also help lessen medical expenses. Results of
the study may add to the list of health benefits and knowledge of Tawa-Tawa as herbal
medicine.
 TABLE OF CONTENTS

PRELIMINARIES
Title Page………..............................................................................................................I
Abstract………….............................................................................................................II
Acknowledgement...........................................................................................................III

CHAPTER I: THE PROBLEM AND ITS SETTINGS

Introduction.......................................................................................................................1
Research Objectives……..................................................................................................2
Hypotheses.......................................................................................................................3
Significance of the Study..................................................................................................4
Scope and Limitation of the Study....................................................................................5
Conceptual Framework....................................................................................................6
Review of Related Local Studies.....................................................................................7
Review of Related Foreign Studies..................................................................................8
Definition of Key Terms....................................................................................................9
References……………………………………………………………………………………10

CHAPTER II: METHODOLOGY

Research Design……………………………………………………………………………...12
Process (Gantt Chart)…………………………………………………………………….13-14
Materials and Methods…………………………………………………….………….…15-17
Research Instrumental Use…………………………………………………………………..18
Safety Pre-Cautions………………………………………………………………………..…19
APPENDICES
Appendix A: Table of Comparison for the Total Cost of Product Made vs.
Commercial.....................................................................................................................20
Appendix B: Research Materials, Ingredients, and Equipment......................................21
Appendix C: Table of Data Collected from Test and Trials…………………....………….22
Appendix D-H: Procedures of Test and Trial……………………………………………23-27
Biographical Data………………………...........................................................................28
ACKNOWLEDGEMENT

The researcher would like to express her genuine appreciation to the following people
and institutions for showing their untiring support to make the study possible:

Grade 9 Research Adviser, Mrs. Lorraine A. Volosso, for guiding this proposal from
the start to completion.

Grade 8 Research Adviser, Mrs. Jane Crystal B. Bayas, for extending her patience
and time in giving feedbacks to this research to make it more informative and objective;

Grade 7 Research Adviser, Mr. Mark Joseph Bautista, for introducing the researcher
to the field of scientific research when grade 7.

Governor Ferrer Memorial National High School, for offering the Science Class
curriculum, which includes, among others, lessons and trainings that expose us to
standards and techniques in conducting scientific and academic researches;

Papa Emmanuel C. Salinas and Mama Roselle Rea G. Salinas, for always guiding the
researcher especially in writing the manuscript and preparing for the defense;

Kuya Xyris Immanuel G. Salinas, for instilling in the researcher the value of listening to
advises and following instructions; and

The Almighty God, for blessing the researcher with knowledge and strength enough to
plan, conduct, and finish the study.
 CHAPTER I

THE PROBLEM AND

ITS SETTING
 INTRODUCTION

Our skin plays a big part on the internal and external health of people and other
living things. Being the largest organ of the human body, it serves as coat and protective
shield to the muscular and internal organs. The skin protects the body against possible
direct adverse effects of chemicals, dirt, air pollution, UV rays, and others. Functional as
it is, but it is also the first organ to be affected by injuries.

One common injury is an open wound, which may have been experienced by
everyone at least once. Most open wounds, though, are minor and can be treated at
home. An open wound is an injury involving an external and internal break in the body
tissues, usually involving the skin. Falls, accidents with sharp objects, and mild burns are
its most common causes (Begum, 2019). While there are a variety of over-thecounter
medicines as remedies to minor wounds, certain herbal plants are said to be potential
alternatives in terms of healing benefits and affordability.

Although most Filipino consumers nowadays seem to prefer synthetic and

commercial medical products over natural and organic ones, elder folks still believe in the
effectiveness and safety of herbal medicines. Tawa-Tawa or Euphorbia Hirta of the
Euphorbiaceae family is one of the most common and well known herbal plants that is
said to counter several health conditions such as dengue and fever.

The possible beneficial use of this herbal plant is further explored, specifically on
its leaves, the pharmacological properties of which include antiseptic, anti-inflammatory,
and anti-bacterial (Tuhin, 2019) among others – all of which as elements to open wound
remedy.

This study, therefore, aims to validate the possible treatment benefits of Euphorbia Hirta
(Tawa Tawa) leaves extract on open wounds.

1
 RESEARCH OBJECTIVES

This study will attempt to prove that Euphorbia Hirta (Tawa Tawa) leaves extract is a
possible treatment for minor types of open wounds.

1. To know if Euphorbia hirta (tawa tawa) Leaves Extract can be an effective treatment
for wounds.

2. To compare which is more useful and effective between comercial product and the
extract.

3. To know the medical use of Euphorbia Hirta (Tawa Tawa) leaves.

2
 HYPOTHESES

1. What pharmacological properties are present in Euphorbia hirta (Tawa Tawa) leaves?
Ho: There are no pharmacological properties present in Euphorbia Hirta (Tawa Tawa).

Ha: There are pharmacological properties present in Euphorbia Hirta (Tawa Tawa).

2. What types of minor open wounds may be treated by applying Euphorbia Hirta (Tawa
Tawa) leaves extract?
Ho: No minor open wounds may be treated by applying Euphorbia Hirta (Tawa Tawa)
leaves extract.

Ha: Certain types of minor open wounds may be treated by applying Euphorbia Hirta
(Tawa Tawa) leaves extract.

3. How effective is the Euphorbia Hirta (Tawa Tawa) leaves extract in treating minor open
wounds compared to commercial ointment?

Ho: Euphorbia Hirta (Tawa Tawa) leaves extract is not as effective in treating minor
open wounds as the commercial ointment.

Ha: Euphorbia Hirta (Tawa Tawa) leaves extract is as effective, if not more effective,
in treating minor open wounds as the commercial ointments.

3
 SIGNIFICANCE OF THE STUDY

Results of this study may beneficial to the following:

The Researcher and the Students. This study may add knowledge to the researcher on
the composition and possible uses and benefits of Euphorbia Hirta (Tawa Tawa) as a
medicine in treating wounds.

The Community. Results of this study may be used to establish among residents the
proven open wound-healing benefits of Euphorbia Hirta (Tawa Tawa) leaves exctract. It may
also serve as basis for opening livelihood opportunities to rural communities.

Medical Researchers. This study may also be added as one of the references in the
medical field on the possible utilization of cheaper and more economical treatments for
open wounds.

The Future Researchers. Others areas related to this study may be explored by future
researchers.

4
 SCOPE AND LIMITATIONS OF THE STUDY

The study entitled, Antibacterial Effect of Euphorbia hirta (Tawa Tawa) Leaves
Extract on Minor Open Wounds, will focus only on the possible healing benefits of the
herbal plant on minor open wounds. Only the leaf part of the plant will be used in the
extraction of juice.

The experiment will be conducted at the researcher’s residence in Buenavista III,

City of General Trias, Cavite, while the variables and materials will be collected from the
researcher’s immediate surroundings. Trial application of the created Tawa Tawa extract
ointment will be done on albino mice. The planning and development, conduct, results, and
manuscript of the study will cover the months of May, June, and July 2021.

5
 CONCEPTUAL FRAMEWORK PROCESS

OUTPUT

Figure 1. Conceptual framework of the study.

Figure 1 presents the conceptual framework of the study, Antibacterial effect of

Euphorbia Hirta (Tawa Tawa) Leaves Extract in Treating Wounds.

The input shows the preliminary procedure for the experiment. It specifically identifies
the variables and materials needed in the study.

Process involves the experiment proper which is the extraction of juice from Euphorbia
Hirta (Tawa Tawa) leaves extract and it’s conversion into ointment.

Lastly, Output includes three possible results from the experiment: production of
ointment from Euphorbia Hirta (Tawa Tawa) leaves extract, The experiment would then
involve the comparison of product created with another commercial product in terms of
healing effect by applying both to wounded laboratory albino mice.

6
 REVIEW OF RELATED LOCAL STUDIES

As stated in the article of Jamie Eske (2019), An open wound is any inside or outside
damage that takes off inner tissue uncovered to the external environment. Individuals can
some of the time treat intense open wounds at domestic with solutions and normal
remedies. However, individuals should look for prompt restorative attention for extreme
wounds that include critical bleeding or broken bones. Wounds disects into two wide
categories: open or closed. In a closed wound, tissue harm and bleeding happen beneath
the surface of the skin. Cases of closed wounds include bruises. An open wound includes a
break within the skin that takes off the inside tissue exposed.
Open wounds may result from falls, blunt injury, and surgery.

Based on the study entitled, Wound healing effect of Euphorbia hirta linn.

(Euphorbiaceae) in alloxan induced diabetic rats, written by Riazul Tuhin,…, Rayhana Begum
(2017), Euphorbia hirta linn.is significant among indigenous people as they consider it a
medicinal plant especially for wounds. Several healing properties are said to be contained this
plant. The study is an evaluation of the healing property of this plant for diabetic animals
through oral and topical application of its ethanol extract. The researchers focused on the
ethanolic extract of E. hirta to find the total phenolic and flavonoid contents with galic acid and
quercetin as basis. First, the rats were induced with diabetes through a solution. Wound sizes
were also determined to for the supposed healing assessment. For sixteen days, the diabetes-
induced rats were given oral and topical ethanol from E. hirta and standard ointment. Then,
then the healing period and mechanisms of the two groups were analyzed and compared.
Investigations showed that wounds healed faster with the E.hirta ethanol extract administered
orally and topically than with diabetic control medicines. Oral E. hirta extract also helped reduce
the glucose content on wounds. The study concluded that E. hirta has a potential for healing
wounds faster than standard iodine treatment among diabeticcontrolled rats.

7
 REVIEW OF RELATED FOREIGN STUDIES

Illustrated on the study of Julie Roddick (2018), An open wound is an harm including
an outside or inside break in body tissue, more often than not including the skin. About
everybody will encounter an open wound at a few point in their life. Most open wounds are
minor and can be treated at home. Falls, mishaps with sharp objects, and car mischances
are the foremost common causes of open wounds. Within the case of a serious accident,
you ought to look for immediate medical care. This is often particularly genuine in case
there’s a part of dying or on the off chance that dying endures for more than 20 minutes.

As written on the research of Linfa Huang and Shilin Chen (2012), A bibliographic
examination was carried out by gathering and analyzing the recognized books, counting
Chinese home grown classic, the Web (Google Researcher, Baidu Researcher), and logical
databases (Pubmed, Scifinder, Scopus, CNKI, ACS, and Web of Science) for accessible data
on Euphorbia hirta. Entirety plants of E. hirta have been utilized in conventional Chinese
medication (TCM) due to its properties that can remedy and detoxify fever, advance diuresis,
halt tingling, and actuate prolactin. Conventional employments of E. hirta are moreover
recorded in a few other pan-tropical regions, where it has been utilized for over 10 sorts of
infections. A number of chemical constituents, counting flavonoids, terpenoids and phenols,
have been disconnected and distinguished from E. hirta.

Research of Linfa Huang and Shilin Chen (2012), Different natural assessments have
been detailed for its significance. Advanced pharmacological examination illustrated that its
unrefined extricates and dynamic compounds has wide pharmacological impacts, such as
antibacterial, anti-inflammatory, antioxidant, narcotic and anti-allergy. The current ponder
summarizes the upgraded data concerning the ethnopharmacology, phytochemistry, and
pharmacology of E. hirta as well as its toxicology, and examines the existing issue and future
inquire about course of E. hirta.

8
 DEFINITION OF KEY TERMS

The following vocabulary will be used and defined operationally in the study as follow:

Anti-Bacterial. This is anything that destroys bacteria or suppresses their growth or their
ability to reproduce. Heat, chemicals such as chlorine, and antibiotic drugs all have
antibacterial properties (www.medicinet.com).

Euphorbiaceae. They are a large family of flowering plants also called the spurge family. In
common English, they are sometimes called euphorbias (Wikipedia.com)

Euphorbia Hirta (Tawa Tawa). Also known as “gatas-gatas,” it is a hairy herb grown in
open grasslands, roadsides and pathways. This indigenous plant is considered as one of
the most popular folkloric treatment for dengue in the Philippines. Tawa-tawa is also able
to promote cell production, and prevents platelet destruction (www.pchrd.dost.gov.ph).

Herbal Medicine. It is the use of plants to treat disease and enhance general health and
wellbeing. Herbs can interact with other pharmaceutical medications in the prevention of
diseases, disorders, and the promotion of good health (www.betterhealth.vic.gov.au).

Minor Open Wounds. This wounds typically do not need medical treatment; they are small
and sufficiently clean so that the edges can be easily approximated using tape
(studenthealth.uiowa.edu).

Over-the-Counter Medicines. They are also known as OTC or nonprescription medicines. All
these terms refer to medicine that you can buy without a prescription. They are safe and
effective when you follow the directions on the label and as directed by your health care
professional (www.fda.gov).

9
 REFERENCES

De Guzman, G. Q., Dacanay, A. T., Andaya, B. & Alejandro, G., (2016, June),
Ethnopharmacological studies on the uses of Euphorbia hirta in the treatment of
dengue in selected indigenous communities in Pangasinan (Philippines). PMC
National Library of Medicine National Institute of Health. Volume 5(3),
10.5455/jice.20160330124637. Retrieved from
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4927128/

Eske, J. & Luo, E. (2019, May), What to know about open wound care.
Medical News Today.
Retrieved from: https://www.medicalnewstoday.com/articles/325260

Huwang, L. & Chen, S. (2012). Euphorbia hirta (Feiyangcao): A review on its

ethnopharmacology, phytochemistry and pharmacology. academicjournals.org.
Volume 6(39) pp. 5176-5185 .
Retrieved from:
https://academicjournals.org/journal/JMPR/articleabstract/AC1AA6A20760

Kumar, S., Malhotra, R. & Kumar, D., (2010, January), Euphorbia hirta: Its chemistry,
traditional and medicinal uses, and pharmacological activities. PMC National Library of
Medicine National Institute of Health. Volume 4(7), 10.4103/0973-7847.65327.
Retrieved form:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3249903/#:~:text=Eup
horbia%20hirta%20is%20often%20used,%2C%20digestive%20probl
ems%2C%20and%20tumors

Roddick, J & Gonzales, A. (2018, September), Open Wound. healthline. Retrieved from:
https://www.healthline.com/health/open-wound

Tuhin, R., Begum, M., Rahman, S., Karim, R., Begum, T., Ahmed, U. S., et. al. (2017, August),
Wound healing effect of Euphorbia hirta linn.
(Euphorbiaceae) in alloxan induced diabetic rats. PMC National
Library of Medicine National Institute of Health. Volume 17,
10.1186/s12906-017-1930-x Retrieved from:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5571567/

10
 CHAPTER III

METHODOLOGY

11
 I. RESEARCH DESIGN

This Research used the Descriptive - Experimental design. Experimental research is a

scientific approach to research, where one or more independent variables are
manipulated and applied to one or more dependent variables to measure their effect on
the latter. The effect of the independent variables on the dependent variables is usually
observed and recorded over some time, to aid researchers in drawing a reasonable
conclusion regarding the relationship between these 2 variable types
(https://www.formpl.us/blog/experimental-research).

In this study, the independent variable was the E. Hirta Tawa-Tawa leaves extract,
and the dependent variable was the wounds of the Laboratory Albino Mice. The effect of
E. hirta leaves extract on the wounds of laboratory albino mice over time were observed
and recorded by the researcher.

Figure 2: Table of Research Outline

12
 II. GantChart/Process

ACTIVITIES
DURATION (DECEMBER – MAY 2022)

W1 W2 W3 W4 W5 W6 W7

1. Planning and development of

the research proposal
(Chapters I,II and III)

2. Collection of variables:
Euphorbia Hirta (Tawa Tawa)
leaves, water, laboratory albino
mice

3. Preparation of materials:
mortar and pestle, casserole, and
others

4.Conduct of experiment on
Euphorbia Hirta (Tawa Tawa)
leaves:

Juice extraction, juice to

ointment conversion, and
Tawa Tawa ointment and
commercial ointment trial on
albino mice with open
wounds

13
 Gantt chart, cont.

DURATION
ACTIVITIES

W1 W2 W3 W4 W5 W6 W7
MAY MAY MAY JUNE JUNE JUNE JUNE 27-
16-22 23-29 30JUNE 6-12 13-19 20-26 JULY 3
5

5. Presentation of possible
results:

Euphorbia Hirta (Tawa

Tawa) leaves in ointment
form

6. Application of
Euphorbia Hirta (Tawa
Tawa) leaves ointment
as home remedy to
wounded albino mice

7. Healing of minor
wounds after application
of Euphorbia Hirta (Tawa
Tawa) leaves ointment.

8. Presentation of the
results and evaluation of
the experiment.

14
 III. MATERIALS AND METHODS

INTRODUCTION ABOUT THE PROCEDURE

To be able to assess the Antibacterial Property of Tawa Tawa (Euphorbia Hirta)

Leaves Extract as Treatment for Wounds, the researcher will perform the presented
methodology below. The procedure dissects into 4 phases which are the collection of the
materials needed, the process of Extraction of Tawa-Tawa leaves, creating the extract
from the Tawa-tawa leaves and comparison of created product and commercial products.
Safety preparations shall be observed in the experimentation proper.

A. GATHERING OF THE MATERIALS

In order to achieve the desired output, the researcher will gather all materials
needed such as measuring cup, syringe, utensils or tools, containers, and different
additives that will help.

Image 1. Collection of Image 2. Collection

Tawa-Tawa leaves of Laboratory Albino Rats

15
B. PROCESS OF EXTRACTION OF TAWA-TAWA LEAVES
For the first process, the researcher will collect 10 pieces about 20 grams of Tawa-
Tawa leaves. from General Trias. Next, start pounding the Tawa-Tawa leaves using
Mortar and Pestle.

Image 3. Weight of Image 4. Pounding of

Tawa-Tawa leaves Tawa-Tawa leaves

C. MAKING THE OINTMENT

After pounding the Tawa-Tawa leaves, the researcher will proceed to the second
procedure, Prepare the crushed Tawa-Tawa leaves. Start heating the 2 cups of water in
casserole. After that, add the Tawa-Tawa leaves to the boiling water, stir the boiling juice
for 1 minute. Next, after stirring the juice for another 1 minute, turn off the stove and filtrate
the extract using a strainer. Next, transfer the extract to a container and let it sit for 20
minutes. After that, The researcher will now proceed to comparison of products.

Image 7. Extraction of Image 8. E. Hirta

Image 5 & 6. Boiling of Tawa-Tawa Leaves
Tawa-Tawa juice Wound Treatment

16
D. TEST AND TRIALS
For the trial proper, the researcher prepared the created extract, commercial
product, laboratory equipment such as Laboratory gown, safety goggles, gloves and the
variable: 5 wounded albino mice and other additives that may help the experimentation.

Proper preparation on the respondents was observed before the Test and Trials.
The experimentation was divided into 2 phase. 1st phase was the semi-trial to know the
right concentration of E. Hirta wound treatment. 2nd phase was the final trial to test
which is more effective in treating open wounds between the created product and
commercial product. The researcher will apply the products on the first 3 Laboratory
rats by gentle swabbing to the external wound.

Image 9. Preparation Image 10. Preparation to Image 11. Testing

to the Laboratory the Laboratory Albino Rats of E. hirta extract
Albino Rats (trial1)

Image 12. Testing of Image 12. Testing of

E. hirta (trial2) E. hirta (tria3)

17
RESEARCH INSTRUMENTAL

The Research Questionnaire was an adapted 5-likert scale research instrument from a
related study concerning in healing stages of open wounds.

18
 PRECAUTIONS

For the experimentation proper, the researcher will utilize safety measures at all times for
the sake of safety while conducting the study. May the posed list guide

1. Clean and disinfect all laboratory/ kitchen materials to before and after using.

2. Before and after activities, keep pathways and the workplace clear. Make sure that
no items can be stepped on.

3. Disinfect hands or any body part that will be exposed to different laboratory
equipment with soap, water and alcohol.

4. Wear safety laboratory gown, gloves, face mask, goggles, closed footwear, and
the hair must be covered with a hairnet.

5. Don’t ever attempt to apply the extract to your external skin.

6. No food and drink are allowed in the workplace.

7. Dispose the waste properly after the experimentation.

19
LIST OF APPENDICES
APPENDIX A: TABLE OF COMPARISON FOR TOTAL COST MADE VS.

20
APPENDIX B: RESEARCH MATERIALS, INGREDIENTS, AND EQUIPMENT

21
22
APPENDIX D

23
APPENDIX E

24
APPENDIX F

25
APPENDIX G

26
APPENDIX H

27
 BIOGRAPHICAL DATA

The Author, Xandrea Romeen Gasilao-Salinas, is a Grade 8 learner in the

Governor Ferrer Memorial National High School-Special Science Class. She finished
her elementary education WITH HONORS at the Evangel Christian Educational Center
in Buenavista III, City of General Trias, Cavite.

Xandy has an active student-life. She was one of the members of the ECEC Kids Choir-
her elementary school choir that earned several provincial championships. Her athletic
skills in swimming was first honed when she was in Grade 3. She became a swimmer in
the ECEC Ychthus Swim Team and experienced competing and winning in various inter-
school, inter-city, and inter-province meets.

In her young age, she is already exposed to community activities such as

Christmas gift-giving and community pantries. She is an active member of the Buenavista
Evangelical Church. She hopes to use soon her talent in playing the guitar and yukelele
in the Praise and Worship Team of the church.

She was born on May 11, 2007 in Trece Martires City. She lives in Buenavista III,
City of General Trias, Cavite. She is the youngest child of Emmanuel C. Salinas and
Roselle Rea Gasilao-Salinas, and has one brother, Xyris Immanuel G. Salinas.
28