Republic of the Philippines

Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

Effectiveness of Peppermint (Mentha piperita lamiaceae) Extract on Pain

Tolerance in C. Elegans Assay

Researchers:
DEANG, EDREYAN M.
ESCAROS, ROWANALEI ATHALIAH A.
GONZALES, LIAM HAILEY D.
LEGASPI, CARL ANGEL E.
MALLARI, JOSEMARI SANTINO V.
RIVERA, YURIKO
SALONGA, PAUL ADDY J.
VALDEZ, JAMES MARCUZ D.

ADVISER
ARNOLD MARAMAG

NOVEMBER 2022
 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

A. The following are the objectives of our study:

1. This study aims to test the efficacy of the Peppermint (Mentha piperita lamiaceae)

extract on Pain Tolerance by utilizing C. Elegans Assay.

2. This study aims to evaluate if Peppermint (Mentha piperita lamiaceae) extract using C.

Elegans will help aid individuals with low pain tolerance.

B. The following are the significance of our study:

The findings of this study is to investigate the effects of Peppermint (Mentha piperita

lamiaceae) extracts that can benefit individuals that are suffering from pain tolerance by making

use of C.elegans assay.

Thus the findings of the research will be beneficial to the following:

● The Researchers. This study will benefit the researchers seeing that it will serve as an

additional knowledge that may be used for their future research journey. This will also

provide them with experience in creating their future studies.

● Department of Health. As one of the major departments that coordinates with

health-related services, this study will give more insights about how Peppermint (Mentha

piperita lamiaceae) extract affects living organisms in terms of pain tolerance.

● Community. This study will give information and knowledge that turmeric Peppermint

(Mentha piperita lamiaceae) extract can be used on living organisms and how much

concentration is safe for humans to take.

 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

● Medicinal. In terms of medicine, this research will benefit a large number of individuals,

including those who aren't impacted by pain tolerance, because it has a variety of health

advantages.

● Agriculture: This study can help in the field of agriculture. Farmers will have more ideas

to plant more Peppermint (Mentha piperita lamiaceae) because they are aware that

Peppermint (Mentha piperita lamiaceae) has benefits.

● Future Researchers. This could serve as a starting point for further research. They could

experiment with new variables and settings to have a better understanding and gather

additional data.

C. The following are the scope and limitations our study:

This study's primary objective is to Test the Effectiveness of Peppermint (Mentha

piperita lamiaceae) Extract on Pain Tolerance in C. Elegans Assay. The variables to be

encountered from the experiment are: the independent variable; varying concentrations of

peppermint (Mentha piperita lamiaceae) extract. The dependent variables to be measured are the

lifespan/mortality rate, chemosensory/behavior, phytochemicals; flavonoids and ph level, and

color. Lastly, the constant variables are Caenorhabditis Elegans without treatment, temperature,

and C. Elegans life span (18-20 days). This study will hopefully be implemented at a laboratory

and at DOST San Fernando Pampanga for the phytochemical analysis of Peppermint (Mentha
 Republic of the Philippines
Department of Education
Region III
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Olongapo City

piperita lamiaceae) to determine the bioactive substances present in these plants. We researchers

are hoping that by January 2023 we will have finished our research proposal and started to gather

materials for the plant identification in DOST San Fernando, Pampanga. The phytochemical

analysis of peppermint (Mentha piperita lamiaceae) and the extraction were completed by

mid-January to early February. The whole month of March for the Caenorhabditis elegans and

the collection and analysis of data until April. We hope that by May 2023 we can submit the final

research output for presentation. Effectiveness of Peppermint (Mentha piperita lamiaceae)

extract on Pain Tolerance in Caenorhabditis Elegans Assay is an experimental study that includes

a hypothesis, a variable that can be manipulated by the researcher, and variables that can be

measured, calculated and compared. This study will not focus on other problems that are not

necessarily connected to pain tolerance and Peppermint (Mentha piperita lamiaceae) extract and

this study will focus at most on the scope of the study only. The C. Elegans Assay will only be

the subject of this study. The main subject for this study would be the C. Elegans Assay but it

will also include the following: scientists, farmers, agriculturists, medicinal, future researchers,

and especially individuals that are suffering from low pain tolerance. This study will undergo the

Gathering of Materials, Plant Identification, Phytochemical Analysis, Extraction, and collection

of data and analysis.

 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

D. We have checked the library resources and found the following related literatures:

Complete Title of Books, Article Complete Entries: Author, This is useful

Titles, Journals, Letters, Internet because….
sources, etc. Name of the book, article,
etc., date, volume, number,
pages

1. “Finding Function in Novel Targets: Kaletta, Titus, and Michael A study that aims at the

C. Elegans as a Model Organism.” Hengartner. “Finding function of C. Elegans

Nature Reviews Drug Discovery. Function in Novel Targets: as a model organism.

C. Elegans as a Model Several features of C.

Organism.”21 Apr. 2006 elegans make it a

powerful tool for the

pharmaceutical industry.

2. “Effects of Emotion on Pain Carter, Leslie, et al. This study focuses on

Reports, Tolerance and Physiology.” “Effects of Emotion on Pain behavioral testing,

Pain Research and Management. Reports, Tolerance and relating to the result of

Physiology.” 2002 emotions leading to

different kinds of pain.

Pain tolerance is tested

connected to the result

 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

of emotions such as

depression and anxiety.

3. “Caenorhabditis Elegans as a Model Ayuda-Durán, Begoña, et It is about a study about

Organism to Evaluate the Antioxidant al. “Caenorhabditis Elegans Caenorhabditis Elegans

Effects of Phytochemicals.” PubMed as a Model Organism to which evaluates the

Central (PMC) Evaluate the Antioxidant antioxidant effects of

Effects of Phytochemicals.” phytochemicals. This

13 July 2020 study is useful for our

study since we will also

be using the C. Elegans

as a Model Organism.

4. “The Mechanisms of Pain Cimpean, Alena, and This study focuses on

Tolerance and Pain-related Anxiety in Daniel David. “The how pain

Acute Pain.” Sage Journals. Mechanisms of Pain catastrophizing and state

Tolerance and Pain-related anxiety affect pain

Anxiety in Acute Pain.” 29 tolerance and anxiety

Nov. 2019, associated with pain.

 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

5. “Phenolic Profile and Antioxidant Dorman, Damien, et al. In relation to our

Evaluation of Mentha X Piperita L. “Phenolic Profile and current study, this

(Peppermint) Extracts.” Phenolic Antioxidant Evaluation of research also focuses on

Profile and Antioxidant Evaluation of Mentha X Piperita L. the Peppermint extract

Mentha X Piperita L. (Peppermint) (Peppermint) Extracts.” in which we will also be

Extracts. Sage Journals. Phenolic Profile and using for our study, it

Antioxidant Evaluation of also gives more insights

Mentha X Piperita L. on how we can use the

(Peppermint) Extracts, 1 Peppermint Extract.

Apr. 2009
 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

6. “Phytochemical Analysis and Pramila, et al. The current study

Antimicrobial Potential of Methanolic “Phytochemical Analysis intends to evaluate the

Leaf Extract of Peppermint (Mentha and Antimicrobial Potential Mentha piperita, or mint

piperita: lamiaceae).” Journal of of Methanolic Leaf Extract plant, available locally,

Medicinal Plants Research. of Peppermint (Mentha leaf extract's

Piperita: Lamiaceae).” phytochemical content,

Academic Journals, 16 Jan. antioxidative activity,

2012 and antibacterial

activity. The

phytochemical

components of the

methanolic leaf extract

of mint leaves were

qualitatively evaluated.

7. “Efficiency of Ethanolic Extract of Nanekarani, Shahram, et al. This study was

Peppermint (Mentha Piperita) as an “Efficiency of Ethanolic conducted to examine

Antibiotic Growth Promoter Extract of Peppermint the effect of different

Substitution on Performance, and (Mentha Piperita) as an levels of ethanolic

Carcass Characteristics in Broiler Antibiotic Growth Promoter extract of peppermint

 Republic of the Philippines
Department of Education
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Schools Division of Olongapo City
Olongapo City

Chickens.” Asian Pacific Journal of Substitution on (Mentha piperita) in

Tropical Biomedicine. Performance, and Carcass drinking water in

Characteristics in Broiler comparison with an

Chickens.” Science Direct, antibiotic growth

2012 promoter

(Virginiamycin) on the

growth performance and

carcass traits of broiler

chicks.

8. “Effects of Emotion on Pain Carter, Leslie, et al. A leading hypothesis is

Reports, Tolerance and Physiology.” “Effects of Emotion on Pain that there is a reciprocal

ScienceDirect. Reports, Tolerance and relationship between

Physiology.” Pain Research pain and mood, which is

and Management, 2002 expressed both in terms

of covariation of "state"

variables and in

diagnostic comorbidity
 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

9. “The Mechanisms of Pain Cimpean, Alena, and This study emphasizes

Tolerance and Pain-related Anxiety in Daniel David. “The the role of pain

Acute Pain.” Sage Journals Mechanisms of Pain catastrophizing and state

Tolerance and Pain-related anxiety on pain

Anxiety in Acute Pain.” tolerance and

Sage Journals, 29 Nov. pain-related anxiety.

10. “The C. Elegans Model in Toxicity Hunt, P. R. (2016, July 22). This article reviews C.

Testing.” Analytical Science Journals. TheC. elegansmodel in elegans use in various

toxicity testing. Journal of toxicity assays, the C.

Applied Toxicology, 37(1), elegans model's

50–59. strengths and limitations

https://doi.org/10.1002/jat.3 for use in predictive

357 toxicology, and GCeCP.

E. The following are the theoretical topics or concepts in Peppermint (Mentha piperita

lamiaceae) that we will be using in the study:

1. Bioactive potential of peppermint extract against some human bacterial and fungal

pathogens.
 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

2. Efficiency of ethanolic extract of peppermint (Mentha piperita) as an antibiotic growth

promoter substitution on performance, and carcass characteristics in broiler chickens.

3. Effects of peppermint (Mentha piperita L.) alcoholic extract on carbon

tetrachloride-induced hepatotoxicity in broiler chickens under heat stress condition

4. Phytochemical analysis and antimicrobial potential of methanolic leaf extract of

peppermint (Mentha piperita: Lamiaceae).

5. The effect of oral Peppermint (Mentha piperita lamiaceae) extract on Migraine.

F The following are the materials, source, quantity, and cost needed for (my/our) study:

Materials Needed Source (where and what Quantity of Cost

company can you materials

purchase) including (grams/liters/cc)

delivery time specify units

Caenorhabditis Elegans DOST, San Fernando, 20-30 worms 1,035.00

Pampanga
 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

Peppermint (Mentha Sm Downtown 10 grams 500

piperita lamiaceae) Olongapo

Ethyl Alcohol Sm Downtown 3 bottles 250

Olongapo

Small Plastic Container Sm Downtown 2 pieces 150

Olongapo

Total Cost of Material(s): 1,935.00

Where is the budget coming from? Researcher’s Initiatives

G. The following are the equipment needed for my/our study:

 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

Equipment Needed Source (where and Quantity of Cost (Lab fees)

what lab can you materials to be

avail of the test) tested

Phytochemical Analysis of DOST, San 1 1,466.00

Peppermint (Mentha piperita Fernando, Pampanga

lamiaceae) extract

Microscope DOST, San 1 0

Fernando, Pampanga

or in a Laboratory

Total Cost: 1,466.00

Where is your budget coming from? Researcher’s Initiatives

 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

H. The following are the summary of the procedure of the study:

The researchers will collect 10 grams of Peppermint (Mentha Piperita Lamiaceae) from

Olongapo, Zambales. After collecting, the researchers will begin preparing water and sanitized

containers for extraction. The collected Peppermint (Mentha Piperita Lamiaceae) will be

gathered and delivered to the Bureau of Plant Industry in San Fernando, Pampanga for plant

identification. The extract of Peppermint (Mentha piperita lamiaceae) will be sent to the DOST

for phytochemical investigation to determine the bioactive substances present in these plants.

The researchers will extract Peppermint (Mentha piperita lamiaceae) using a juicer, and the

extract will be transferred on a small plastic container with a lid. The extracts will be tested on C.

elegans and will be conditioned and observed under the microscope. After 10 minutes,

peppermint extracts will drop once on the C. elegans and will be observed afterwards. Once the

testing is done, the researchers will collect and analyze data that was tested from the peppermint

extracts that was drop from the C. elegans

I. The following are the schedule of activities in doing our the study (from start to
finish):

The whole first grading was filled with planning and working on our research starting

from the title to the other parts and ending on the background of the study. The research process

started in early September and finished specifically on October 24-26, 2022. Early on September

2022, Individually, we researchers started looking for a research that would definitely help a

common and famous problem. A lot of students picked Robotics, Physical science, and also life
 Republic of the Philippines
Department of Education
Region III
Schools Division of Olongapo City
Olongapo City

science. On September 19, we gathered as a group to look forward to inventing something out of

nothing. Starting September 21, we finalised 3 choices of title from the group to look further

what research title would fit everyone’s likes and interests. The next day, September 22, all of the

groupmates agreed to use Turmeric on pain tolerance as our research topic for this school year to

develop. September 24, the title and topic that we chose are disregarded and rejected. We are told

to research more for alternatives of turmeric. September 25, we chose peppermint as the

alternative for turmeric. On this day discussed, our title has been approved. Finalising the title

and began to provide Independent, Dependent, and Constant Variables of this study, September

26, 2022. After providing the amount of variables to use for the study, we are asked to research

more, to look for further variables to improve our research study, September 27. On the next day,

September 28, our compiled and completed variables were finally approved. On the same day,

we are finally taking our next step to create and finalise our research hypothesis, yet it is not

finished on the same day so the remaining time. We used free and vacant times of our weekend

to create and finalise our hypothesis. Yet not everything succeeds in just once or twice. On

October 3, our hypothesis got rejected and was told to revise and renew. Our hypotheses are

finally revised, researched for more specific variables to count adding improvements on our

research study. In between the days of October 4 to October 10, we focused on finalising our

statement of the problem, hypothesis. Also the same day October 10, we focused on editing and

finishing our tarpaulin for the research exhibit and parade on the next day. October 11, the parade

started at 7am and the exhibit started at 9am. On this day, our tarpaulin is displayed outside the

setting of the exhibit. Research symposium happens to be held on October 12 in the afternoon.

And on October 13-21, our hypothesis, variables, and statement of the problem were finished.
 Republic of the Philippines
Department of Education
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Schools Division of Olongapo City
Olongapo City

We also had 1-2 paragraphs to finalise our background of the study. And finally on October

24-26, the 1st chapter of our research study has been finished, which consists of the background

of the study, statement of the problem, conceptual framework, hypothesis, and variables.

In the 2nd quarter, we started working on starting and completing the significance of the

study of this research, November 3. This part states what are the benefits of our study to an

amount of people. November 4, the action plan was discussed to everyone and how is it done and

planned according to each research study. On the same day, we finalised our research

significance of the study using each member’s works. Everyone contributed and we also

finalized our scope and delimitation by each member’s work to complete and finish the work on

the same day, 4th of November.

We hope to complete all of the steps and processes required for a successful research

study by mid-2023, or before the school year ends.