Anti-Nociceptive Activity of Muntingia Calabura Linn Leaves (Aratiles) Using Methanol Extract On Swiss Albino Mice

ANTI-NOCICEPTIVE ACTIVITY OF MUNTINGIA CALABURA LINN LEAVES…
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Anti-nociceptive Activity of Muntingia Calabura Linn Leaves (Aratiles) using

Methanol Extract on Swiss Albino Mice
Josuah D. Alvarez123, Patrick James Aquino123, Raffy R. Arizapa123, Cary Mae A.

Cayanan123, Sophia Kaye P. Pangilinan123, 123, Winsher May C. Talbo123, Winslet May C.
Talbo123,, Jojin Grace N. Tarriman123, Trisha B. Valeriano123, Nemuel Jabez M. Yaco123,
John Paolo J. Angeles1234, LPT, Brylle T. Capili1234, RN
1
Senior High School
2
Basic Education Department
3
Our Lady of Fatima University
4
Research Adviser
March 2019
OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

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Endorsement
This research paper entitled “Anti-nociceptive Activity of Muntingia calabura linn Leaves
(Aratiles) using Methanol Extract on Swiss Albino Mice” prepared by Winsher May Talbo
et al. of STEM 12YI-HEA-1 of Our Lady of Fatima University, in partial fulfillment of the
requirements for Senior High School has been examined and now recommended for Oral
Examination.
This is to certify that Winsher May Talbo et al. are ready for the Oral Examination.
Brylle T. Capili, RN
Adviser
This is to certify that the research paper, “ Anti-nociceptive Activity of Muntingia calabura
linn Leaves (Aratiles) using Methanol Extract on Swiss Albino Mice” prepared and
submitted by Winsher May Talbo et al. of STEM 12YI-HEA-1 is recommended for Oral
Examination.
Chair
Member Member
Dr. Ernesto B. Guevarra, MD, MPH, MSTM
Chairman, Biology Department
Senior High School – Research Coordinator

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Certificate of Originality
We hereby declare that this research paper is our own work and that, to the best of our knowledge
and belief, it contains no material previously published or written by another person nor material
to which to a substantial extent has been accepted for award of any other degree or diploma of a
university or other institute of higher learning, except where due acknowledgement is made in
text.
We also declare that the intellectual content of this research paper is the product of our work,
even though we may have received assistance from others on style, presentation and language
expression.
Winsher May Talbo

Principal Investigator
Members:
Josuah D. Alvarez
Patrick James Aquino
Raffy R. Arizapa
Cary Mae A. Cayanan
Sophia Kaye P. Pangilinan
Winsher May C. Talbo
Winslet May C. Talbo
Jojin Grace N. Tarriman
Trisha B. Valeriano
Nemuel Jabez M. Yaco
Brylle T. Capili, RN
Adviser
Date

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Table of Contents
Endorsement ii
Certificate of Originality iii
Table of Contents iv
Abstract 1
1.0 Introduction 2
1.1 Statement of the Problem 3
1.2 Research Hyphotesis 4
1.3 Scope and Limitations 4
1.4 Significance of the Study 4
1.5 Definition of Terms 5
1.6 Conceptual Framework 6
2.0 Review of Related Literature 7
2.1 Foreign Literature 7
2.1.1 Nociceptive Pain 7
2.1.2 Muntingia Calabura Linn 8
2.1.3 Phytochemical Analysis of Muntingia Calabura Linn Leaves 9
2.2 Local Literature 10
2.2.1 Nociceptive Pain 10
2.2.2 Muntingia calabura Linn 10
3.0 Research Methodology 10
3.1 Research Materials 11
3.1.1 Animals 11
3.1.2 Collection of Muntingia Calabura Linn Leaves 11
3.2 Research Procedure 11
3.2.1 Extraction of Muntingia Calabura Linn Leaves 12

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3.2.2 Anti-nociceptive Activity 12
3.2.2.1 Chemical Test (Formalin-Induced Paw Licking Test) 12
3.2.3 Statistical Analysis 12
References 13
Appendix
A- Certificate of Authentication 17
B- Receipt of Methanol and Formalin 18
C- Institutional Animal Care and Use Committee 19
D- Permission to use Rotary Evaporator 24
List of Figures
Figure 1 - The Conceptual Framework 6

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Abstract
Nociceptive pain refers to pain clearly associated with tissue damage or inflammation
(Loeser and Treede, 2008). It is caused by potentially harmful stimuli being detected by
nociceptors around the body. Results from stimulation of pain receptors for tissue injury
(nociceptors), which are located mostly in the skin or in internal organs. The injury may be a cut,
bruise, bone fracture, crush injury, burn, or anything that damages tissues.
The use of natural product as alternative therapies is an increasingly popular method to

treat such discomfort, either alone or as a complement to conventional medical approaches.
Muntingia calabura L. (family Elaeocarpaceae) has been traditionally used to relieve various
pain-related ailments in the Southeast Asia and tropical America to treat headaches and gastric
ulcer, tranquillizer and anti-nociceptive.
The in vivo chemicals (formalin-induced paw licking test) models of nociception were
used to assess the extract anti-nociceptive activity. The extract (10, 5, and 400 mg/kg) was
administered orally 60 min prior to subjection to the respective test. The study gives quality to
Muntingia calabura Linn leaves and uses it as an inhibitor for nociceptive pain.
Keywords: Nociceptive pain, Muntingia Calabura linn leaves, Elaeocarpaceae

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1.0 Introduction
One of the most common reason of discomfort is pain. According to National

Survey on Drug Use and Health, (2013 & 2014), an annual average of 10.7 million
people aged 12 or older used prescription pain relievers in the past year. Pain may be
sharp or dull, intermittent or constant, or throbbing or steady. Pain may be felt at a single
site or over a large area. The intensity of pain can vary from mild to intolerable.
Nociceptive pain is the most common type of pain. Nociceptive pain arises from various
kinds of trouble in tissues, reported to the brain by the nervous system. This is the type of
pain everyone is most familiar with, everything from bee stings and burns and toe stubs
to repetitive strain injury, nausea, tumors, and inflammatory arthritis. Nociceptive pain
typically changes with movement, position, and load (Ingraham, 2019). It is caused by
potentially harmful stimuli being detected by nociceptors around the body. It develops
when the nociceptive nerve fibers are triggered by inflammation, chemicals, or physical
events, such as stubbing a toe on a piece of furniture (Weatherspoon, 2017).
There are three major classes of nociceptors in the skin: mechanosensitive

nociceptors, mechanothermal nociceptors, and polymodal nociceptors, the latter being
specifically associated with C fibers (Purves et al., 2013). In C-fibers, there are thought to
be three mechanisms that activate its nociceptive pathway: thermal, mechanical, and
chemical (Cavanaugh, 2009). The nociceptive tests for thermal and chemical stimulation
are well characterized and standardized in both rats and mice (Barrot, 2012). Tests using
thermal and chemical stimuli, measures of hyperalgesia and allodynia, models of
inflammatory or neuropathic pain, constitute a toolbox available to researchers. These
tests and models allowed rapid progress on the anatomo-molecular basis of physiological
and pathological pain, even though they have yet to translate into new analgesic drugs
(Barrot, 2012).
The use of natural product as alternative therapies is an increasingly popular

method to treat such discomfort, either alone or as a complement to conventional medical
approaches (Wirth et al., 2005). According to Holt and Chandra (2002), categorized
natural products as herbs, herbal concoctions, traditional Chinese medicines, dietary
supplements and alternative medicines. The research in natural product based on its ethno

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pharmacological knowledge has provided substantial contributions to drug innovation

through the discovery of novel chemical structure and/or mechanism of actions (Rates,
2001).
Muntingia calabura linn, the sole species in the genus, has been traditionally used
in the Southeast Asia and tropical America to treat headaches and gastric ulcer,
tranquillizer and anti-nociceptive (Kaneda et al., 1991). Various medicinal properties
have been scientifically reported from Muntingia calabura leaves, including anti-tumor
(Su et al., 2003), hypotensive (Shih et al., 2006), antibacterial, antiplatelet aggregation
(Chen et al., 2007), anti-inflammatory, antipyretic and anti-nociceptive (Zakaria et al.,
2007). And one of the plants and leaves are reported to possess high anti-inflammatory
and antioxidant activities and as a potential source of hepato-protective compounds is
Muntingia calabura linn. Based on the literature review on Muntingia calabura for
pharmacological reports between 1991 and 2014, it has been reported to demonstrate
cytotoxic, anti-proliferative, insecticidal, hypotensive, anti-nociceptive, cardio-protective,
antipyretic, antiplatelet aggregation, antioxidant, anti-inflammatory, antidiabetic,
antiulcer, and antibacterial activities with approximately 87 flavonoid-based compounds
identified (Mahmood et al., 2014).
The aim of this research is to know the level of anti-nociception of Muntingia

calabura leaves. It will be tested in male Swiss Albino mice (20-30g; 8-10 weeks old) to
examine the viability of Muntingia calabura linn as an anti-nociceptive and natural
alternative inhibitor for nociceptive pain.
1.1 Statement of the Problem
The main problem of the study is to determine if the methanol extract of

Muntingia calabura linn (Aratiles) leaves has an anti-nociceptive effect against chemical
tests of nociception.
The study aims to answer the following:
1. Is the methanol extract of Muntingia calabura linn leaves an effective inhibitor of

nociceptive pain?

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2. In what phase of time is the methanol extract of Muntingia calabura linn leaves more
effective?
3. Is the methanol extract of Muntingia calabura linn leaves a more effective inhibitor
than the analgesic Morphine?
1.2 Research Hypothesis
This study seeks to examine the viability of using the methanol extract of
Muntingia calabura linn (Aratiles) leaves as an alternative inhibitor for nociceptive pain
and to conclude these hypotheses:
Ha: Muntingia calabura linn leaves methanol extract is an effective inhibitor for
nociceptive pain.
Ho: Muntingia calabura linn leaves methanol extract is not an effective inhibitor
for nociceptive pain.
1.3 Scope and Limitations
This study centers only on the examination of the methanol extract of Muntingia
calabura leaves (MEMC) on Swiss Albino mice using in vivo chemical tests of
nociception. The researchers will use the methanol extract of Muntingia calabura
(MEMC) to reduce pain on Swiss Albino mice to elucidate the anti-nociceptive activity
of MEMC. Any efforts made that are beyond framework and the aim of the study is
otherwise consider as separate to the whole study. The determination of toxic and lethal
doses of the extract of Muntingia calabura linn including its adverse effect will not be
determined in the study.
1.4 Significance of the Study
The study gives quality to Muntingia calabura Linn leaves and uses it as an
inhibitor for nociceptive pain. Since nociceptive pain is a worldwide issue, the use of
Muntingia calabura Linn leaves methanol extract as an alternative pain reliever could
help people more if it was proven effective or not. If effective, a more natural and cheap
treatment will be discovered. If not effective, it will be a step for the improvement and

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exploration of a new drug for nociceptive pain. This paper will uncover the possible anti-
nociceptive activities of Muntingia calabura Linn leaves as well as the mechanisms
involved.
Community. If the methanol extract of Muntingia calabura Linn is proven to be

effective in this research, the community will gain knowledge about this plant to engage
in making solutions connected with nociceptive pain, and to use this as an alternative for
prescription pain reliever.
Government. This research can be used as a starting point for the government to use this
plant as a cheaper and alternative herbal medicine not only for nociceptive pain but to
start finding its other benefits as well.
Future Researchers. It will serve as their guide and can act as a reference in their
research. It will give them an idea to know what to do as the researchers answers
questions and produce data and information. This will also serve as an awareness for the
possible problems the future researches may encounter.
1.5 Definition of Terms Used
Analgesics – These are medicines that are used to relieve pain.

Anti-nociception - The process of blocking the detection of a painful stimulus by
sensory neurons.
In vivo - An experiment that is done in vivo is done in the body of a living organism as
opposed to in a laboratory method that does not use the living organism as the host of the
test
Methanol - Is an ideal solvent because it can extract both hydrophilic and lipophilic
molecules from plant parts and after extraction, the solvent can be removed at low
temperature by distillation as methanol is highly volatile.
Nociceptive pain – Used to describe the pain from physical damage or potential damage
to the body.

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Nociceptors –It can be defined as sensory receptors that are activated by noxious stimuli
that damage or threaten the body's integrity.
1.6 Conceptual Framework
A representation is provided to depict the actual process to be done in the research

investigation. Inside it, three sections are provided namely; the input, the process, and the
output, all of which provides a brief insight with what materials are invested into the
study, what process is made to those materials, and what is the output that was
manufactured afterward, respectively. The representation is as follows:
Collection and extraction of Muntingia calabura

linn leaves
Acclimation of Sprague Dawley rats
Administration of respective test solutions to

rats
Testing the anti-nociceptive acitivity of Muntingia

calabura linn leaves using the Hot-Plate Test and
Formalin-Induced Paw Licking Test
Collection and analysis of data
Fig 1. The Conceptual Framework

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2.0 Review of Related Literature
This section of the research is allotted for the previous known information about
different variables in the study. Unfortunately, since the research is severely understudied
in the country, the researchers had a difficult time finding a relevant local literature
regarding the subject matter. Although there are few, but still the amount compared to the
foreign literature used in the study is unfortunate.
2.1 Foreign Literature
These literatures are closely related to the current study for it directly mentions
the benefits of Muntingia calabura linn leaves that specifically provides medicinal cases
for the said study. The present study like the article reviewed will help researchers to
provide a cure for nociceptive pain.
2.1.1 Nociceptive Pain
The International Association for the study of pain defines pain as an unpleasant
sensory and emotional experience associated with actual or potential tissue damage or
described in terms of such damage. It recognizes that tissue damage does not need to be
present for pain to be experienced (Nicholson, 2006). They appear to respond to many of
the same forms of noxious stimuli that activate cutaneous nociceptors and also participate
in neurogenic inflammation (Gebhart, 2000). Specialized peripheral sensory neurons
known as nociceptors alert us to potentially damaging stimuli at the skin by detecting
extremes in temperature and pressure and injury-related chemicals, and transducing these
stimuli into long-ranging electrical signals that are relayed to higher brain centers (Invest,
2010). The changes in nociceptive processing that occur during illness are believed to be
mediated partly by descending facilitation of spinal nociception. These pro-nociceptive
effects most likely represent the hyperalgesia noted in the many studies of illness in
awake animals (Anthony, 2013).
More recently, a growing effort has been put forth trying to assess pain in rats or
mice, rather than nociceptive reflexes, or at studying states affected by chronic pain
(Barrot, 2012). It is generally accepted that the activation of primary afferent C-fibers by

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noxious stimuli leads to a sensation of pain (Cho, 2016). The close correspondence
between nociceptor properties and human pain perception has been confirmed using a
variety of experimental approaches (Marks, 2006). And also, nociceptive pain continues
only as long as the noxious stimulus is maintained. Sustained or recurrent noxious
stimulation can occur in certain disease. As a result, the nociceptive to control of
behavior routinely occurs in the absence of consciously perceived pain rendering it
"subconscious" (Meyer, 2006).
2.1.2 Muntingia calabura Linn
Muntingia calabura linn is a tropical plant species that belongs to the

Elaeocarpaceae family (Mahnmood et al., 2014). It is a fast growing tree that is
widespread and growing along roadside. It can reach up to 40ft.tall at full maturity. The
soft leaves are evergreen and have an alternate phylloxytacy. Leaves are dark green on
top and light green underneath (Sarojini&Mounika, 2018). Muntingia calabura has been
traditionally used by the Peruvian to treat various ailments (Balan, 2012). The Muntingia
calabura leaves exhibited potential anti-proliferative that is used to prevent perilous cell
around the tissues and antioxidant activities. (Sarojini&Mounika, 2018). Cherry leaves or
Muntingia calabura leaves contains anti-inflammatory and anti-nociceptive that generally
form by phenolic or pholifenols, the sinamat acid derivatives, flavonoids, tocopherols,
coumarin and polifungsional acids (Sani, 2012).
Leaves can either be boiled or steeped in water to provide relief from gastric ulcer
or to reduce swelling of the prostate gland, while the strips of its bark are boiled and
washed to reduce the swelling in the lower extremities. The leaves have been used to treat
pain associated with gastric ulcers, headache, and cold or to depress the prostate gland
swelling (Zakaria et al., 2007). In addition to that, the leaves were used as antiseptics or
antipruritic; also, to treat abdominal cram and also used to relieve colds and headaches in
provinces (Stuart, 2016). Studies have shown antioxidant, anti-inflammatory, anti-
pyretic, analgesic, cytotoxic, hepatoprotective, gastroprotective, cardioprotective,
antibacterial, antiulcer, insecticidal, tyrosinase-inhibitory, Antispasmodic, emollient and
antifungal properties (Stuart, 2016).

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Methanol, petroleum ether, and ethyl acetate are the solvents used for plant
extraction of phytochemical analysis including formic acid, and LCMS grade acetonitrile
(Selangor, 2019). The way to separate desired substance when mixed together is called
Extractions (Triswaningsih et al., 2017).
2.1.3 Phytochemical Analysis of Muntingia calabura linn Leaves
Phytochemical analysis of various leaf extracts of Muntingia calabura linn leaves

yielded saponin, tannins, triterpene, steroid, and flavonoids. LC-MS analysis of leaves
yielded fumaric acid (6.643%), succinic acid (4.903%), niacin (0.718%), malic acid
(2.863%), cinnamic acid (4.945%), pyridoxine (1,893%), gallic acid (21,428%), ascorbic
acid (6.121%), glucose (8.166%), fructose (20.690%), pantothenic acid (1.478%), biotin
(1.025%), thiamin (1.158%), kaempferol (6.825%), catechin (14.407%), quercetin
(10,623%), riboflavin (1.131%), and folic acid (1.553%)(Ramasamy, 2017).GC-MS
analysis of leaves for volatile compounds yielded geraniol (26.335%), eugenol
(19.950%), citronellol (16.958%), α-amyrin (6.225%), myrcene (3.440%), and α-
terpineol (7.356%). LC-MS analysis for phenol compounds yielded gallic acid
(18.607%), catechin (14.077%), quercetin (10.255%), ellagic acid (9.626%), and
kaempferol (8.699%) (Buhian, 2016).Study of stem wood yielded a new biflavan, (M),
(2S),(2''S)-,(P),(2S),(2''S)-7,8,3',4',5',7'',8'',3''',4''',5'''- decamethoxy-5,5'' biflavan (1), a
new flavone, 4'-hydroxy-7,8,3',5'-tetramethoxyflavone (2), and a new dihydrochalcone,
(R)-2',β-dihydroxy-3',4'-dimethoxydihydrochalcone (3), together with 12 known
compounds (Kumalaningsih, 2017).
Various solvent extracts (water, methanol, ethanol, chloroform, ether) yielded

bioactive constituents saponin, tannin, and flavonoids. Qualitative phytochemical
analysis of methanol solvent yielded epigallocatechin gallate and genistein of 135.15
µg/g and 135.29 µg/g, respectively. (30) GC-MS analysis of leaves for volatile
compounds yielded myrcene (5.927%), thymol (3.543%), a-terpinol (11.831%), linalool
(2,240%), geraniol (21.718%). nerol (4.375%), citronellol (12.837%), eugenol
(17.498%), α-lonone (1.413%), ß-sitosterol (7.806%), α-amyrin (3.167%), lupelol
(4.228%). α-tocopherol (1.975%) dan ß carotene (1.425%) (Ramasamy, 2017).

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2.2 Local Literature
The researchers found it challenging to gather local literature about the research
topic. Thus this topic is severely understudied in the country. Fortunately, phytochemical
analysis show properties that make researchers assume that the Muntingia calabura linn
leaves will have a possible impact to inhibit nociceptive pain.
2.2.1 Nociceptive Pain
In the Philippines, analgesic and anti-inflammatory medications may be

appropriate, it is important to understand the underlying etiology (Gomez, 2009). It is
also experienced when the somatosensory nervous system exhibits any kind of abnormal
function (i.e. nociceptive pain) (Santos, 2009). The Philippines has a relatively high
prevalence of nociceptive pain. There is a need for improved pain management to ensure
that patients with nociceptive pain receive appropriate and effective treatment (Henry,
2009).
2.2.2 Muntingia calabura Linn
Aratiles is a fruit which grows naturally anywhere within the Philippines (Zulueta,
2019). Muntingia calabura linn leaves shows potential anti-proliferative and antioxidants
activities that could be accredited to their high contents of phenolic compounds. The
leaves also apply potent anti-tyrosinase and antioxidants activities (Maria et al., 2008).
Also in the Philippines, the flowers are used to treat headache, relief of incipient colds,
antiseptics and to treat swelling in the lower extremities (Cruiz, 2016).
3.0 Research Methodology
This section of the research paper is allotted for the description of the
methodologies involved in the study. Divided into two subsections, the research materials
and research procedures are listed below along with a brief description provided for each
subsection.

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3.1 Research Materials
These are the materials that the researchers utilized to know if the methanol
extract of Muntingia calabura linn leaves may inhibit nociceptive pain in Sprague
Dawley rats.
3.1.1 Animals
9 male Swiss Albino mice were divided into 3 groups. On each group, there are 3
mice that weighs between 20-30g ages 8-10 weeks old and which the researchers bought
in the Bioresearch at SM North EDSA Annex which is accredited by the Bureau of
Animal Industries Philippines. They were kept under room temperature (27 ± 2°C; 70–
80% humidity; 12 h light/darkness cycle) in the Pet-link Wellness Center and Veterinary
Clinic in Victoneta Avenue, Malabon City. The animals had free access to food and
water. They were fasted overnight before the experiment. The animals were acclimatized
to the laboratory conditions for at least five days prior to the experiments. The mice were
at all times handled in accordance with current BAI guidelines for the care of laboratory
animals and the ethical guidelines for investigations of experimental pain and conscious
animals (Zimmermann, 1983). Furthermore, experiments (n=3) were conducted between
09.30 and 18.30 h to minimize the effects of environmental changes.
3.1.2 Collection of Muntingia calabura Linn Leaves
The Leaves of Muntingia calabura are collected from within Valenzuela City.
The sample was tested in the UP Biology Institute of Biology College of Science
(University of the Philippines) for identification and certification purposes.
3.2 Research Procedures
Research is an orderly and systematic procedure, and these are the following
procedures that the researchers followed in order for the experiment to be successful and
have a reliable outcome.
3.2.1 Extraction of Muntingia calabura linn leaves

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This procedure was carried out as described in detail by Zakaria et al. (2011).
Briefly, 500g of matured leaves that have been air-dried for 2 weeks at room temperature
(27 ± 2°C) and grinded into powder will be soaked in methanol in the ratio of 1: 20 (w/v)
or 10 L for 72 hours. After that, the supernatant is filtered using steel filter, cotton, and
What man no. 1 filter paper. The residue subjected to the same procedures for another
two times. The supernatant collected from each extraction pooled together and then
subjected to evaporation process using a rotary evaporator at 40°C under reduced
pressure.
3.2.2 Anti-nociceptive Activity
Anti-nociception activity is the process of numbness in a certain part or nerve in

our body, it blocks the detection of pain or painful stimuli. It tests encompass a large
group of experimental procedures specifically developed for examining sensitivity to
painful stimuli and the alteration of pain sensitivity following drug administration.
3.2.2.1 Chemical Test (Formalin-Induced Paw Licking Test)
The formalin test was carried out as described by Pharmacol et al. (2011) but with
slight modifications. Pain was induced by injecting 20 μL of 1% formalin in the
subcutaneous under dorsal surface of the right hind paw. Controls received distilled water
(10 ml/kg), the positive group received 100 mg/kg Tolfenamic acid as the reference drug,
and the experimental group received MEMC (500 mg/kg), 60 min prior to the formalin
injection. Immediately after the formalin administration, the mice was individually place
in a transparent glass cage observation chamber. The amount of time the animal spent
licking their injected paw, was considered as an indicator of pain, and was be recorded
for a duration of 30 min in two phases, known as the early (0–5 min) and late (15–
30min) phases.
3.2.3 Statistical Analysis
The statistical analysis of Sani et al. (2012) will be followed wherein the results
will be presented as Mean ± standard error of mean (SEM). The one-way ANOVA test

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with Dunnett post hoc test is used to analyze and compare the data, with P < 0.05 as the
limit of significance.
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17
Appendix A
Certificate of Authentication

18
Appendix B
Receipt of Methanol and Formalin

19
Appendix C
Institutional Animal Care and Use Committee

20

21

22

23

24

25
Appendix D
Permission to use Rotary Evaporator

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ANTI-NOCICEPTIVE ACTIVITY OF MUNTINGIA CALABURA LINN LEAVES…

Anti-nociceptive Activity of Muntingia Calabura Linn Leaves (Aratiles) using

Josuah D. Alvarez123, Patrick James Aquino123, Raffy R. Arizapa123, Cary Mae A.

OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

Dr. Ernesto B. Guevarra, MD, MPH, MSTM

Chairman, Biology Department

Senior High School – Research Coordinator

OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

Winsher May Talbo

OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

Certificate of Originality iii

1.1 Statement of the Problem 3

1.2 Research Hyphotesis 4

1.3 Scope and Limitations 4

1.4 Significance of the Study 4

1.5 Definition of Terms 5

1.6 Conceptual Framework 6

2.0 Review of Related Literature 7

2.1 Foreign Literature 7

2.1.1 Nociceptive Pain 7

2.1.2 Muntingia Calabura Linn 8

2.1.3 Phytochemical Analysis of Muntingia Calabura Linn Leaves 9

2.2 Local Literature 10

2.2.1 Nociceptive Pain 10

2.2.2 Muntingia calabura Linn 10

3.0 Research Methodology 10

3.1 Research Materials 11

3.1.2 Collection of Muntingia Calabura Linn Leaves 11

3.2 Research Procedure 11

3.2.1 Extraction of Muntingia Calabura Linn Leaves 12

OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

3.2.2 Anti-nociceptive Activity 12

3.2.2.1 Chemical Test (Formalin-Induced Paw Licking Test) 12

3.2.3 Statistical Analysis 12

B- Receipt of Methanol and Formalin 18

C- Institutional Animal Care and Use Committee 19

D- Permission to use Rotary Evaporator 24

Figure 1 - The Conceptual Framework 6

OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

The use of natural product as alternative therapies is an increasingly popular method to

Keywords: Nociceptive pain, Muntingia Calabura linn leaves, Elaeocarpaceae

OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

One of the most common reason of discomfort is pain. According to National

There are three major classes of nociceptors in the skin: mechanosensitive

The use of natural product as alternative therapies is an increasingly popular

OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

pharmacological knowledge has provided substantial contributions to drug innovation

The aim of this research is to know the level of anti-nociception of Muntingia

1.1 Statement of the Problem

The main problem of the study is to determine if the methanol extract of

The study aims to answer the following:

1. Is the methanol extract of Muntingia calabura linn leaves an effective inhibitor of

OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

1.2 Research Hypothesis

1.3 Scope and Limitations

1.4 Significance of the Study

OUR LADY OF FATIMA UNIVERSITY SENIOR HIGHSCHOOL

Community. If the methanol extract of Muntingia calabura Linn is proven to be