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1.0 Introduction

Sudden death can be considered as ones most frightening phobia, unfortunately, this is just
a symptom of a disorder called pulmonary thromboembolism that any individual can have with just
a simple blood clotting or coagulation. According to Sharma & Golhani (2017) pulmonary
embolism, stroke, and myocardial infarction, which are all leading cause of death, are all caused
by the ailment thrombosis (blood clot). Aside from surgical intercession or bypassing the blockage,
the only treatment left is injecting thrombolytic agents to dissolve the coagulation.

Some thrombolytic agents that are widely used worldwide such as streptokinase (SK), tissue
plasminogen activator (t-PA), and urokinase (UK) have a damaging and harmful side effects, that
is why scientist and biochemist searched for a more natural way substitute for those harmful
thrombolytic agents (Sharma & Golhani, 2017), and that is where the concept of the researchers
came in. Capsicum frutescens widely known by the locals as Siling labuyo, has proven to contain
phytochemical and anticoagulant properties making it a credible substitute for the harmful
thrombolytic agents thus maximizing the usage of Siling labuyo and promoting herbal medicines.

Capsicum frutescens linn is a member of Solonaceae family. It is a very popular additive in

cooking. Most commonly for giving culinary dishes its fiery flavor. Aside from its common uses,
Siling labuyo has also some herbal uses. It increases blood flow, induces sweating, reduce muscle
tension and spams and relieves gas and colic. Recent studies also suggest its anticoagulant and
antioxidant properties.

Anticoagulants achieve their effect by suppressing the synthesis or function of different

coagulating factors normally present in the blood. Such medications are used to stop the blood clots
from developing in the veins or lungs or to prolong a clot circulating throughout the bloodstream.
A blood clot formed in the bloodstream due to hemostasis failure causes artery blockage and
recovery leads to severe effects in atherothrombotic diseases including acute myocardial or cerebral
infarction, often resulting in death (Anwar et al., 2011).

Two main study groups the test samples and the samples under supervision. The subjects
of the research are the persons used to control what happens when something changes.
Effective control is part of a constructive control experimental design. A positive control is given
a proven reaction care in order to compare this positive response with the uncertain response of the
treatment. This result is usually what researchers learn through therapy, it is used in an experiment

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to track unknown factors and to provide the researcher with something to correlate with the
experimental group.

The control test is essential as it allows the researcher by comparing results to determine
the baseline of the experiment. Traditional control materials are designed to resemble samples of
plants and are checked with samples of plants to determine the aspect of the analysis. Positive
controls are considered to be reactive, and negative controls for the analyst being tested are non-
reactive. To properly evaluate the test performance, the controls should have the same structure, or
matrix, as patient samples, including viscosity, turbidity, and color. A positive control is an
experiment involving the use of effective medication to replicate the test. It is a replicate experiment
that enables the researcher to confirm the accuracy of the test results.

1.1 Statement of Problem

This study aims to determine the effectiveness of Capsicum Frutescens (Siling Labuyo) as an
alternative anticoagulant.

The researchers aimed to answer the following questions:

1. Is there a significant difference between positive control group and experimental group
treated samples on microscopic examination of red blood cell morphology, micro
clumping and microhematocrit?

2. Is there a significant difference in the anti-coagulating effect between the positive

control group and experimental group treated with Capsicum frutescens extract?

3. Is Capsicum frutescens extracts an effective anti-coagulant in vitro?

1.2 Hypothesis

H1: Capsicum Frutescens Linn (Siling Labuyo) exhibits an alternative for anti-coagulant, and
the effect will be measured by in vitro activity.

H0: Capsicum Frutescens Linn (Siling Labuyo) does not exhibit an alternative for anti-
coagulant, and the effect cannot be measured by in vitro activity.

H0: There is no difference in the Capsicum Frutescens Linn (Siling Labuyo) as an alternative
for anti-coagulant.

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1.3 Significance of the Study

 Allied Professions
This gives details regarding the anti-coagulating properties of the Capsicum
frutescens components.
 Students
This study will provide them with some knowledge and help them become
knowledgeable about this study.
 Community
The study may provide evidence that will increase community understanding of
Capsicum frutescens ' efficacy as an anti-coagulant, providing a more cost-effective
tool for blood collection.
 Future Researcher
This study will serve as a guide for the other researches to use this as a basis for
future studies.

1.4 Scope and Delimitation

All experiments were performed in the laboratory of Our Lady of Fatima University by the
Head department of the Valenzuela Campus and use with the Health's Guide for the Care and Use
of the experiment instrument.

The research sample is composed of Capsicum frutescens that contained a wide range of
nutritional components and pharmacologically active metabolites. Moreover, the researcher
positive control group is the heparin that acts to prevents the formation of blood clots. To establish
the process, we will test if there is a present agent in the Capsicum frutescens that can prevent blood
clots.

This study is limited to its scope in assurance of the adequacy of the anticoagulant property
of Capsicum frutescens. The test does exclude data about capsicum separates relating to experiment
does not include information about capsicum extracts pertaining to its other therapeutic effects (i.e.,
antifungal and antimicrobial properties). The extraneous variable will be additionally kept
constrained by keeping the activity as institutionalized as would be prudent, where all activities did
by the researcher were guaranteed to be firmly indistinguishable from one another.

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2.0 Review of Related Literature

2.1 Foreign Literature

2.1.1 Anti-Microbial Activity

There are many economically expensive medications presently in use are not
available in rural regions. Contradictory use of such drugs can contribute to resistance to
microbial drugs. Phytochemicals exert antimicrobial behavior into various mechanisms;
alkaloid has been found in all Capsicum pepper extracts, which is one of the largest groups
of phytochemicals in plants. Their toxicity to foreign organism cells is one of the most
important biological properties of alkaloids. Dorantes et al. (2005), which demonstrated
how Capsicum's effective substances are soluble for both aqueous and isopropanol solvent
extracts and also have the antibacterial capacity.

Extracts collected from Capsicum frutescens display antimicrobial activity

towards certain foodborne disease-causing bacterial pathogens. Alkaloids, flavonoids,
polyphenols, and sterols have been established as pharmacologically active compounds.
Therefore, Capsicum fruits are used in the food and medicinal systems as a source of
natural antibacterial. This article presents information about the use of pepper and is a
justification for the use of Capsicum fruit extracts for antibacterial activity. Such findings
will provide scientific credence to support the use of pepper as an ingredient in food or in
traditional medicine. Incorporating traditional knowledge with scientific findings will
increase community understanding.

2.1.2 Capsaicin

Capsicum frutescens is one of the oldest herbal medicines that have been used for
over 5000 years. The present study indicates that capsaicin's anti-thrombotic role is due not
to any antiplatelet activity but to its effects on the normal and extrinsic pathways. Jolayemi
& Ojewole (2014) observed that capsaicin has thrombolytic effects on heat-sensitive
vanilloid receptors. This was a statement on the study of capsaicin's cardiovascular effects.
Jolayemi & Ojewole (2014) have reported that the frequency of thromboembolism is lower
for individuals who ingested a large amount of Capsicum. Capsaicin is also an active
ingredient of the antithrombotic activity of Capsicum frutescens.

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2.1.3 Effects of Capsaicin on Coagulation

The effect of capsaicin on coagulation was reported in several studies. Current

research is being directed at elucidating if capsaicin has any effect on coagulation. To
investigate if either the ethyl acetate extract of Capsicum frutescens linn capsaicin on
coagulation. The Addition of capsicum to meals may achieve the required
thromboprophylaxis, especially in the population at risk. However, high dose of oral
Capsicum frutescens fruit extract could lead to gastric erosion as seen in this study (Ezekiel
et al., 2014). Taken together, the use of capsaicin at therapeutic doses may reduce
thromboembolism without any clinically relevant alteration in platelets.

2.1.4 Therapeutic properties of medicinal plants

The recent studies showed that many plants possessed cardiovascular effects. This
review was designed to cover the hypolipidemic, fibrinolytic anti-platelet aggregating and
antioxidant effects of the medicinal plants. This paper reviewed the hypolipidemic,
fibrinolytic anti-platelet aggregating and antioxidant effects of the medicinal plants to open
the door for their clinical uses as a result of efficacy and safety (Al-Snafi., 2015). Plants
and their secondary metabolite constituents have a long history of use in modern medicine
and in certain systems of traditional medicine, and are the sources of important drugs such
as atropine, codeine, digoxin, morphine, quinine and vincristine.

2.1.5 Biochemical parameters

Capsicum frutescens it was used for the treatment of diabetes, blood pressure high
or low, bronchitis, burning feet, to increase circulation, relieve rheumatic pain, treat mouth
sores and infected wounds, reduce blood clots, and aid digestion by stimulating saliva and
gastric juice flow (Anthony et al, 2013). So Capsicum frutescens can also use as treatment
for some ailments. Therefore, extracts from Capsicum frutescens has properties that can be
use in the study

2.1.6 Pharmacological importance

The pharmacological importance of capsicum species grown in Iraq. Capsicum

annuum and Capsicum frutescens contained a wide range of nutritional components and
pharmacologically active metabolites (A.E. Al-Snafi, 2015). A wide range of
pharmacological activities was exerted from these two species. The present review will

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highlight the chemical constituents and the pharmacological and therapeutic effects of
Capsicum annuum and Capsicum frutescens.

2.1.7 Varieties of Pepper

Referred to as chili peppers belong to C. annuum L. However, some varieties with

chili included in their name are actually C. frutescens L. Precise categorization of this
particular type of pepper is difficult because of the large number of varieties, and the
constant creation of new ones by hybridization. Forms sold or grown by one name in certain
areas of the country may not be the same elsewhere, even though the names are the same.

2.1.8 Chemical composition of Capsicum fruits

The sharp taste of Capsicum peppers is due to a mixture of seven related alkaloids
of which capsaicin is the most prevalent. Capsaicinoids are mainly found in the seeds and
placental area. The capsaicin content is negligible in sweet bell peppers but found in high
concentrations (30,000 - 50,000 SHU) in hot chili or jalapeno peppers that even handling
or cutting the peppers can irritate the skin (Nwokem et al., 2010). Pepper also contains
carotene, mineral elements, vitamins A and C. Vitamins A, C, and Carotene facilitate the
growth and function of human beings, while minerals help the body withstand stress, cold
and stimulate mucous that protects intestinal lining from ulcer. Minerals, especially trace
elements: iron, manganese, lead, cobalt, chromium, cadmium, zinc and copper are
classified as essential and unessential that are necessary in biochemical reactions that take
place in the body of human beings (Showemimo and Olarewaju, 2004).

2.2 Local Literature

2.2.1 Antimicrobial and medicinal potentials

A previous study showed that capsaicin significantly stimulates the development

of Bacillus subtilis as aqueous and ethanoic pepper extracts are tested for antimicrobial
activity toward Staphylococcus aureus, Bacillus cereus and Bacillus subtilis resistant strain
of Benzylpenicillin (George et al., 2010). Compounds with proven antimicrobial can be
great significance in pharmaceutical drugs and the effective properties were recognized as
alternative simple and effective medicinal drugs toward serious microbial infections (Rojas
et al., 2006). It distinguished through strong organic compounds who are well known for
their roles in pharmacology, neurology, and nutrition. It significantly reduces blood,

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myocardial and aortic total cholesterol levels when used at low levels in normal diets
(Nwokem et al., 2010). So peppers play a vital role in the proper functioning of various
systems within the human body. Therefore, extracts from Capsicum have some useful
antimicrobial activity. Capsicum species ' neuroprotective properties were well
documented.

2.2.2 Hemostatic, fibrinolytic or anticoagulant effects

Compounds insulated from Capsicum frutescens display antimicrobial activity

against certain foodborne disease-causing bacterial infections. As bioactive substances,
alkaloids, flavonoids, polyphenols, and sterols have been described. Therefore, Capsicum
fruits can serve in food and medicinal systems as a source of natural antibacterial. This
article provides data about the use of pepper and will be a justification for the use of
Capsicum fruit extracts for antibacterial action (Kouassi et al., 2012).

Ali Esmail Al-Snafi (2015) has recently used an in-vitro thrombolytic template to
test the effect of Capsicum frutescens on clot lysis. A combination of honey and Capsicum
frutescens and streptokinase as a positive control and water as negative control were
examined. Using an in vitro thrombolytic template, the effect of Capsicum frutescens and
a mixture of honey and Capsicum frutescens was 57.40% and 44.54% respectively. This
included an anticoagulant that helps to prevent heart attacks from the blood clots.

2.2.3 Hepatoprotective activity

Folk remedies from plant origin are tested for their potential anti-oxidant and
hepatoprotective activities. Extract of Capsicum frutescens leaves was obtained through
methanolic extraction which yielded 12.6%. Studies showed that these anti-oxidant
compounds are concentrated in both leaf and fruit extracts of the plant which prevent free
radical damage, reducing the risk of chronic diseases (Soumya et al., 2012). Capsicum
frutescens is an excellent source of various anti-oxidant compounds like flavonoids,
carotenoids and Vitamin C. Capsicum frutescens extract is a therapeutic agent against
hepatotoxicity since it renders lower AST and ALT levels.

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2.2.4 New oral anticoagulants and oculoplastic surgery

The use of oral anticoagulants is increasing because of the rising prevalence of

atrial fibrillation and other cardiovascular disease that occur in an aging population.
Oculoplastic surgeons should work in tandem with an internist to stratify patients according
to their risk for perioperative bleeding and vision loss if anticoagulants are continued (lng
et al., 2014). The use of oral anticoagulants is increasing because of the rising prevalence
of atrial fibrillation and other cardiovascular diseases that occur in an aging population. In
the last decade, several new oral anticoagulants have emerged, comprising the direct
thrombin inhibitor dabigatran and the factor Xa inhibitors apixaban and rivaroxaban.
Although these new anticoagulants may not affect clear corneal cataract surgery, their use
will impact the management of patients having eyelid, orbital, and nasolacrimal
procedures.

2.2.5 Alternative treatment

Capsaicin for medicinal use comes from Capsicum frutescens and is the active
ingredient in the extract of hot peppers. It is most concentrated in the rib or membrane, less
in the seeds, least in the flesh. Capsaicin depletes substance P in the afferent type C sensory
nerve fibers, affecting only proprioception. Unlike other treatments for neuropathy, such
as local anesthetics, opiates, anti-seizure medications or tricyclic antidepressants, capsaicin
specifically treats pain without impairing other aspects of the nervous system. In
incomplete depletion of substance P from suboptimal use, it may cause paradoxical
increase of pain. (G. Stuart, 2014). Therefore, Capsicum frutescens can also be use an
alternative medicine in some ailments because the extract properties came from the
Capsicum frutescens could be very useful in this study

2.2.6 Compounds of Capsicum frutescens

Capsicum frutescens is an excellent source of various anti-oxidant compounds like

flavonoids, carotenoids and Vitamin C. Studies showed that these anti-oxidant compounds
are concentrated in both leaf and fruit extracts of the plant which prevent free radical
damage, reducing the risk of chronic diseases (A.B. Tamayo, 2014). It shows in the study
that there is a lot of nutrient that can be extract from Capsicum frutescens that can be used
in hepatoprotective activity of methanolic extract of Capsicum frutescens against isoniazid
and rifampicin-induced hepatotoxicity.

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2.2.7 Local nomenclature for Capsicum

Two species of Capsicum are used in the Batanes Islands. C. annuum (two pungent
types: round or long fruit type) and C. frutescens (several types: green or greenish yellow
immature fruit color, and several fruit sizes). Unlike C. annuum, C. frutescens is often used
for seasoning foods in the daily diet, and it has become an important and indispensable
condiment in the Batanes Islands. People on the islands of Batan and Sabtang generally
called Capsicum pepper which is probably derived from `chile' in Spanish (Santos, 1988),
and people of Itbayat Island called it kasiri' which seems cognate also.

2.2.8 Plant Specimen and Extract Preparation

Seeds of Piper nigrum and Capsicum annnuum var. glabriusculum are used as
plant specimens to evaluate their antimicrobial activity against Staphylococcus aureus. The
plant parts were collected from Bicol, Naga city in the month of September. The seeds
were cleaned and dried and made into homogeneous powder by grinding. Solvent
extraction method was used to extract the active components from the powder. The plant
extract was prepared as per the protocol powder. The plant extract was prepared as per the
protocol (Pavan, et al.,2010).

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2.3 Conceptual Framework

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3.0 Methodology

3.1 Materials

MATERIALS PURPOSE PICTURE

It contains a compound
called Capsaicin which
guarantees appropriate blood
stream all through the vessels. It
additionally decreases side
effects of poor blood
dissemination like torment,
headaches, numb hands, cold feet
2 Kg of Capsicum
and shivering in legs. They keep
Frutescens Linn (Siling
the platelets from remaining
Labuyo)
together in this manner
facilitating the blood stream. In
addition, Capsicum frutescens
extract is a stronger anti-
coagulant property and it appears
to be useful in platelet
conglomeration and the
movement of clotting variables.

The oven drying

treatment was done using a
laboratory drying oven. CFL was

Laboratory Drying Oven spread out in a single layer on the

shelves and was subjected to
drying at a temperature of 65oC
for 12 hours, with the oven door
left partially open in order to

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prevent cooking. Drying was
carried out from 8am to 4:30pm
daily until the CFL was dry.

The CFL was removed

from the drier at sunset, placed in
plastic crates and stored indoors
overnight. In all three treatments
the CFL was dried until it
obtained a crisp texture and made
a rattling sound when shaken.
The CFL trays were rotated
regularly inside the dryer in order
to ensure that the CFL dried
evenly. The moisture content of
the dried product was recorded.
Moisture content of the CFL was
measured using a moisture
analyzer (MOC63u, Shimadzu
Moisture Analyzer). The
moisture analyzer was set to
measure moisture at a
temperature of 120o C.

Analytical balance it is
use to know the exact measure of
Capsinoids that we need and the

Analytical Balance analytical balance measurement

pan is inside a transparent door
frame so that the dust does not
accumulate and so that any air
currents in the room do not

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impact the function of the
balance.

It is the removal of
solvent in Capsicum frutescens.
Put cold water to the Dimroth
condenser to the collection flask
bath (add ice if needed), clip
Rotary Evaporator
boiling flask with sample in place
and start the rotation boiling flask
with a slowly evacuate system.
Apply heat through the warm
water bath.

The test tube will be use

as a container, after the Rotary
evaporator the extracts from the
Test Tubes Capsicum frutescens will
dispense in the test tubes.

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Test tube rack will be use

to hold the test tubes that contain
extracts from Capsicum
Test Tube Rack frutescens.

Foil it is use to dry the

Capsaicinoids and prevents the
leakage of organic solvents and
Foil will not taint the sample.

Anticoagulant
medication, heparin is significant
in keeping up normal hemostasis
after medical procedure. Heparin
is a sugar like atom that delays
blood clotting by diminishing the
movement of thrombin, a key
segment of the thickening
Heparin component. Heparin acts quickly
however commonly should be
managed parenterally by
intravenous or subcutaneous
courses. Oral anticoagulants take
a few days to influence blood
thickening since they delayed
affect coagulating factor
biosynthesis. Heparin and oral

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anticoagulants regularly are
utilized successively to control
extreme thickening; which can be
regulated for a little while or
months to keep up ordinary
coagulation.

Ethanol is a natural
byproduct of plant fermentation
and also can be produced through
the hydration of ethylene.
Ethanol
Ethanol is used in the
management of toxicity due to
ingestion of methanol, or
ethylene glycol.

A Mortar is a vessel in
Mortar and Pestle which 2kg of Capsicum
frutescens are grind with a pestle.

3.2 Research Design

The researchers will investigate anti-coagulant property Capsicum frutescens in preventing

blood coagulation in vitro.

This study will be using a quantitative experimental research design attempts to establish some
causality between the variables that have been established for this study that focuses on the vitro
activity of Capsicum Frutescens Linn (Siling Labuyo) as an alternative diagnostic anti-coagulant.

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The researchers will use heparin as a positive control. Heparin is an anticoagulant blood thinner
that keeps blood clots from forming. The researchers will follow the procedures, from gathering
the materials through experimentation.

3.3 Research Locale

The study will be conducted at Our Lady of Fatima University. The experiment will take place
at the Laboratory for it is the proper place to conduct experiments. Also, the laboratory of Our Lady
of Fatima University contains complete laboratory tools and equipment that is needed in the study.

3.5 Data Collection

In collection of data the researchers will be focusing in 2 methods first the observation study
and second is the experiment. In observational study the researcher will focus in observation of the
variables of the study which is that Capsicum frutescens. The observation will focus on how the
extract phytochemical and anticoagulant properties of Capsicum frutescens will react before and
after it mix to thrombosis. Next is The experiment, the researcher will focus in extracting the
phytochemical and the anticoagulant properties in Capsicum frutescens. And because the
researcher will be using a vitro activity we will be using laboratory equipment instead of organism.

3.6 Data Analysis

The data will be statistically treated with independent samples t-test using 0.05 level of
significance. The t-value will show if the units of the two groups are apart. There are positive and
experimental control groups that will serve as the foundation of this experiment. Heparin will act
as the positive group, while Capsicum frutescens will be the experimental group. To ensure that
anticoagulation will be comparable to the therapeutic range in Capsicum frutescens, the researchers
will measure the standard coagulation parameters. Standard deviation will be utilized as well. The
data will be calculated using a software called Statistical Package for Social Sciences (SPSS). The
formula for independent samples t-test as follows:

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