CHAPTER I

INTRODUCTION

Background of the Study

Cardiovascular disorders such as hypertension, cerebral hemorrhage, coronary

thrombosis, arteriosclerosis, and congestive heart failure are caused by an abnormality in the

blood circulatory system as blood clotting disorders constitute a serious medical problem. A

blood clot prevents blood from flowing normally through the circulatory system. Regardless of

previously conducted and confirmed research of blood clotting being a necessity, a

life-supporting mechanism, and a good thing in the prevention of excessive bleeding, a blood

clot is most commonly associated with chances of heart attacks and strokes. Hemostasis is the

physiologic process for the prevention of hemorrhage and bleeding in response to blood vessel

damage, while physiologic inhibition of coagulation ensures the fluidity of blood. Platelets and

plasma proteins inside the blood allow this to happen by forming a solid mass over the broken

blood vessel, which can result in a bruise or a scab. When blood clots form inappropriately inside

an artery or vein, they may cause significant knots because blood flow past the formed clot is

decreased, still making it a detriment.

Setaria Palmifolia (more commonly known as palm grass) is a plant that grows in

temperate or tropical countries, like Asia, making it reasonable to be abundant in Ormoc City.
The plant has also been deliberately introduced to further east to Australia, New Zealand, Pacific

Islands and South and Central America.This species is a perennial grass with stems growing up

to two or three meters long from a knotty rhizome and naturally inhabits tropical and subtropical

rainforests. Its leaves have a pleated texture and are hairless or with some rough hairs. The plant

may be dispersed as rhizome fragments but the spread of individual plants by rhizome is very

limited. Most spread is therefore by seeds which are produced quite abundantly. There is little

published, however, on germination requirements or dormancy. Several varieties of the palm

grass are recognized, including some with red foliage. Moreover, the plant is often cultivated for

ornamental purposes, and are usually found in front yard lawns and even in sidewalks.

Polyphenols are micronutrients that can be obtained through certain plant-based foods.

It’s thought that polyphenols can improve or help treat digestion issues, weight management

difficulties, diabetes, neurodegenerative disease, and cardiovascular diseases. Experts believe

that this is largely due to the antioxidant properties of polyphenols, which help reduce chronic

inflammation, a risk factor for heart disease. According to test-tube and animal studies,

polyphenols may help reduce the platelet aggregation process and prevent the formation of blood

clots. Research has provided evidence that polyphenols are able to inhibit the activity of many

enzymes. Setaria palmifolia was 10th in polyphenol content, with a total polyphenols 6.9 + 1.2

(mg/g) (Asian Journal of Chemistry, 2009; Vol 21 No 7: pp. 5556-5562) Since Setaria palmifolia

contains polyphenols, this can be utilized as an anticoagulant. There are no existing studies that

Setaria palmifolia can be an anticoagulant but research journals have stated that it contains
polyphenols that can prevent thrombosis, a formation of a blood clot within a blood vessel that

prevents it from flowing normally.

The researchers chose to conduct this study since it is cost-effective and due to the

aforementioned abundance of “Setaria Palmifolia” in Ormoc City, thus limiting the range of the

study significantly. It would also allow the study to have a lesser chance of having a depleted

source for the necessary research.

Statement of the Problem:

This study aims to verify the potential of Palm Grass (Setaria palmifolia) flower extracts as an

anti-thesis for blood clotting (anti-coagulants).

It specifically aims to:

1. Create an anticoagulant from Pulverized Palm Grass (Setaria palmifolia)

2. Help city folk in terms of:

a. Lesser medical payments

b. Lesser circulatory disease records

 c. Lesser risks of circulatory-related diseases

3. Compare commercial medicinal anticoagulants with our experimental herbal

anti-coagulant in terms of:

1) Effectiveness

2) Cost

3) Accessibility

Hypotheses:

Alternative Hypothesis: 1 out of 4 pulverized and capsuled parts of Palm Grass (Setaria

Palmofilia) as anti-coagulants have a significant difference from the commercial medicinal

anti-coagulants.

Null Hypothesis: None out of 4 pulverized and capsuled parts of (Setaria Palmofilia) as

anti-coagulants have no significant difference from the commercial medicinal anti-coagulants.

Significance of the study:

The significance of this study is to provide and introduce an alternative towards efforts in making

and innovating previous research on non-commercial anticoagulants which are cost effective and

easily accessible.

This study is highly beneficial to the following recipients:

1. People- The findings of this study will redound to its main beneficiary, the individuals

suffering from formation of blood clots within their body. This study will serve as an alternative

non-commercial antithesis/anticoagulant to blood clotting, that is cost-effective and can be

accessed easily.

2. Ormoc City- this study will foster new ways to prevent blood clotting in an affordable way

that requires a small amount of effort and the materials are easily accessible.
Scope and Limitations of the Study

This study is restricted on the utilization of Palm Grass (Setaria Palmifolia) as an antithesis to

blood clotting. Its potential will be measured through an experimental process involving

capsule-making.

Time and Place of the Study

This study will be conducted from this school year 2020-2021. This study will be conducted

within Ormoc City. The specific location of the study will determined by considering the factors

such as, the accessibility of product and the availability of needed materials

Definition of Terms:

Hemostasis

-retarding or stopping the flow of blood within the blood vessels

Anticoagulant
 - preventing coagulation, especially of blood.

Polyphenols

- Polyphenols are micronutrients that we get through certain plant-based foods.

Antithesis

- the direct opposite of someone or something else.

Thrombosis

- a formation of a blood clot within a blood vessel that prevents it from flowing normally.

CHAPTER II

REVIEW OF RELATED LITERATURE

1.1 Related studies and journals

Plants have been known to be sources of medicinal organic treatments. In connection

with coagulation, guava (Psidium guajava) gave a significant difference to the blood count in

ICR mice (Health Research and Development Information Network. Philippines, 2019), this

proves the plant's effectiveness as an anticoagulant. Other studies include Terminalia bellirica,

Astragalus arbusculinus, and Origanum vulgare, as cavulgares for herbal medicines in the
prevention and treatment of cardiovascular diseases, such as heart attacks and strokes (Iranian

Biomedical Journal, 2017).

Recently, a research study conducted by Shikha Jain, CBS Dangi, Manpreet Kaur,

Hardev Singh, Jaya Peter and Sushmit Kosta of World Journal of Pharmaceutical Research

(2014), Palm Grass (Molineria recurvata) leaf extract which specifically contains methanol is

used for its anticoagulant activity on fresh human blood. The extracts were found to have

sufficient anticoagulant activity. But the concentration of 2g/ml methanolic leaf extract showed

the maximum effect with respect to other plants involved in the study with its anticoagulant

activities being observed. It detects deficiencies in factor X. Factor X deficiency, also called

Stuart-Prower factor deficiency, is a condition caused by not having enough of the protein known

as factor X in your blood. Factor X plays a role in blood clotting, also called coagulation, which

helps you to stop bleeding. Inherited factor X deficiency cannot be cured. Thus prolongation

indicates a deficiency.

Similarly, ( Sudan Journal. of Medical Sciences, 2011) also studied red onion (Allium

cepa) for its potential as an anticoagulant. The aqueous extract of red onion was found to inhibit

coagulation process in vitro and significantly prolonged prothrombin time in a dose-dependent

manner. They then concluded that the results showed that red onion aqueous extract in different

concentrations inhibits clot formation and increases prothrombin time. Red onion can be used as

a supplementary anticoagulant agent to improve and/or prevent cardiovascular diseases.

 According to, (CABI) Centre for Agriculture and Bioscience International (2012) Setaria

palmifolia is a densely tufted perennial grass with short, woody, knotty rhizomes, with foliage up

to 1 m high. Individual leaves 20-60 cm long, 2-7 cm, wide, plicate (finely pleated with multiple

ridges) glabrous or hispid, narrowed toward the base, apex acuminate. Leaf sheath hispid with

irritant hairs, ciliate near the ligule which is 2.5-3.5 mm long, also ciliate. The plant is also

widespread across the country and is also used for medicinal purposes. It may be sometimes used

as an ornamental plant for gardens and other decorations.

1.2 Journals that contribute to the potential of palm grass (setaria palmifolia) as an

anticoagulant

Stated in the (Asian Journal of Chemistry, 2009) The contents of polyphenols in 23 dry

plant materials were estimated. The contents of polyphenols in 23 dry plant materials ranged

from 2.5 to 164 mg/g in which top ten was in the order of; Acacia confusa, Macaranga tanarius,

Euphoria longana, Mangifera indica, Ficus wightiana, Lantana camara, Pittosporum pentandrum,

Leucaena leucocephala, Ficus septica and Pittosporum pentandrum. Except for Anredera

cordifolia, Ophiopogon japonicus and Setaria palmifolia.. Polyphenols are micronutrients that

we get through certain plant-based foods. They're packed with antioxidants and potential health

benefits. It's thought that polyphenols can improve or help treat digestion issues, weight

management difficulties, diabetes, neurodegenerative disease, and cardiovascular diseases.

 According to a study conducted by the (International Journal of Health Medicine and

Current Research, 2016) From the test results, it discovered that palm grass (Setaria palmifolia)

contains Nonadecane compound with content of (39,79%) that is potentially an antioxidant.

According to Thomas S.C Li, Ph.D. in his book (Taiwanese Native Medicinal Plants,

2006) Setaria Palmifolia (Koen.) Stapf. S. viridis (L.) Beauv. The root part of the plant contains

tricin,p-hydroxycinnamic acid, vitexin 2"-oxyloside, orientin 2"-o-xyloside,

tricin-7-o-B-d-glucoside, vitexin 2"-o-glucoside. Such contents improve stomach and intestinal

function. It also improves digestion, eating disorder, arthritis, heart disease. Activities in the

plant like free radical-scavenging activities were observed on 1,1-diphenyl-2pierylhydrazyl

(DPPH) of the plant. It was also found that antioxidant contents are contained by the plant.

With the use of the gathered data, the researcher found potential from the palm grass

(setaria palmifolia) as an effective anticoagulant. Setaria palmifolia has been tested and proven

for its antioxidative property (IJHMCR, 2016) and for its active polyphenol content

(Asianjourn,2009). During the last decade many researchers have also provided evidence that

polyphenol compounds are able to inhibit the activity of many enzymes, including serine

proteases. All research described in this review clearly indicate that polyphenols and

polyphenol-rich extracts possess not only antioxidative but also anticoagulant properties and may

be useful in creation of new therapeutic agents or medical supplements (NCBI, 2016). Thus

enabling the researchers to see potential in palm grass (Setaria palmifolia) as an anticoagulant.
 1.3 Studies about blood related illnesses

From a study of the Philippine Council for Health Research and Development of the

year 2012, the prevalence of hypertension among Filipinos is undeniably increasing. A doctor

involved in the study also added that the continuous increase of hypertension cases may be

attributed to continuous neglect on the danger of hypertension and its complications. In this

study the author also stated that hypertension is considered as the biggest single risk factor for

deaths worldwide. According to the World Health Organization (WHO), hypertension causes 7

million deaths every year while 1.5 billion people suffer due to its complications. PhilStar

(2014), stated that based on the calculation extrapolated from the number of thrombosis-related

deaths across Europe, thrombosis or blood clot kills one person every 37 seconds across the

Western world. In a global interview during the same year, only one in 10 people consider blood

clots as life threatening as AIDS, breast cancer, prostate cancer or road traffic accidents. Due to

the stigma, people tend to disregard and not pay full attention to Thrombosis. Currently, recent

studies still confirm the high risk of hypertension and blood related illnesses which mainly

causes fractions of deaths among countries.

WebMD, a medical news and information site suggested that blood clotting may cause problems

in the removal of waste in the body, such problems can cause high blood pressure, blood related

disorders or even kidney failures. With the risk of such illnesses, researchers found its way to

foster treatments about coagulant activities inside the body such as blood clotting.
References:

https://www.jstage.jst.go.jp/article/cytologia/76/3/76_3_309/_pdf

https://www.philstar.com/lifestyle/health-and-family/2014/10/14/1380106/1-person-dies-every-3

7-seconds-deep-vein-blood-clot

https://www.ncbi.nlm.nih.gov/pubmed/5826244

http://www.thepharmajournal.com/archives/2019/vol8issue4/PartR/8-4-69-641.pdf

https://ejournals.ph/article.php?id=13398

https://www.cabi.org/isc/datasheet/119079

https://books.google.com.ph/books/about/Taiwanese_Native_Medicinal_Plants.html?id=VxPNB

QAAQBAJ&redir_esc=y

file:///C:/Users/Ader%20Pepol/Downloads/2014Plantasaantithromboitic%20(1).pdf

https://www.ajol.info/index.php/sjms/article/view/72459
 CHAPTER III

METHODOLOGY

Research Design

This study will use an experimental research design. An experimental research design is the best

research approach in this study. Using such will allow the researchers to control, experiment, and

explore the variety of factors and variables involved within the study that could affect the

production of capsuled pulverized Palm grass (Setaria palmifolia) as an anticoagulant agent.

This design would also enable the researchers to arrive at more valid and accurate results.

Materials:

· 3 2-inch tall; 13×9 inches Baking Pan

· Food Processor

· 4 Bowls

· Butcher’s twine (Cooking twine)

· Paper towel
 · Mortar and pestle

· 100 hole size “0” Manual Capsule filling Machine

· Size “0” (500mg) Hard Gelatin Capsules

· Oven

· 4 Spoons

· 4 Palm grass (Setaria palmifolia) blades

· 6 Palm grass (Setaria palmifolia) sheaths (with maximum length of 7

inches)

· 125 g Palm grass (Setaria palmifolia) seeds

· 125 g Palm grass (Setaria palmifolia) roots

Research Procedures:

A. Preparation of Palm Grass

 1. Collection and cleaning of Palm Grass

Firstly, the researchers will collect Palm grass (Setaria palmifolia) blades, sheaths, roots

and seeds. Once the parts are already collected, the sheaths, along with the blades, seeds,

and roots will be washed thoroughly using only clean water and tied together using

butcher’s twine. Then, the plants will be hung in a cool and dry place and the seeds will

be laid evenly on a 2-inch tall; 13×9 inches baking pan with a paper towel covering the

surface of the pan below and above the seeds, both away from direct sunlight to preserve

its contents. The plants will be left to dry for 1-2 weeks while the seeds for 5-7 days, the

researchers will check the plants and the seeds once a day for signs of mildew and mold.

2. Desiccation of Palm Grass

For an easier alternative, the Palm grass (Setaria palmifolia) blades, sheath, roots and

seeds will be baked in the oven after being hanged and laid out to dry. In this process, the

4 Palm grass (Setaria palmifolia), blades, the 6 Palm grass (Setaria palmifolia) sheath,

125 g of Palm grass (Setaria palmifolia) roots will be laid vertically and evenly while the

125 g of Palm grass (Setaria palmifolia) seeds will be scattered and laid, all on a 2-inch

tall (28 x 18 cm) baking pan separately and will be put inside the oven. The oven will be

set to 200 ºF (93.333 ºC) and the desiccation can begin, the blades will be turned upside

down and sideways every 10 seconds for even drying. When 1-2 minutes pass and the

plants become dry and crisp, the desiccating process ends.

 3. Pulverization of Palm Grass

After desiccating, the blades, sheaths, roots and seeds will be separated into 4 different

bowls. The researchers will pulverize the desiccated blades, sheaths, roots and seeds

separately using a food processor on medium speed initially and increases in the process.

Then, the food processed separated parts of the plants will be transferred and will be

pounded using a mortar and a pestle. Afterwards, the process will be repeated 2-3 times

for a finer product. Then, the pulverized blades, sheaths, roots and seeds will be placed

back into the 4 different bowls.

B. Filling of Capsules

A 100-hole size #00 Manual Capsule-filling Machine will be used in filling the capsules.

Firstly, the halves of the capsules will be pulled apart, leaving the tops aside. The researchers

will place the base of the machine on its stand which secures it in place while filling the capsules

and fusing them together. The base of the machine will be loaded by the longer or bottom halves

of the capsules while the top part of the machine will be loaded by the shorter or top halves of

the capsules. Using a spoon, the researchers will scoop the pulverized blades, sheaths and seeds,

and are placed separately at the base of the machine to fill the holes where bottom halves of the

capsules are loaded. Then, the pulverized blades, sheaths and seeds will be spread into each

bottom half, swinging left and right, back and forth using a plastic card that comes with the
machine to evenly distribute the powder. Afterwards, the researchers will use the tamper in the

machine’s kit to compact the pulverized blades, sheaths and seeds into the capsules. The filling

processing will be repeated until all the bottom halves are fully filled and compacted with the

pulverized blades, sheaths and seeds.

The researchers will line up the top of the machine with the bottom and press down. Then, the

base of the machine will be removed from the stand. The top of the machine will be gently

flipped over so the openings in the top and bottom are lined up with each other. Afterwards, the

top of the machine will be pressed down by the researchers until it stops compressing. At this

point, the capsules will be stored in a cool and dry place to prolong its shelf life. All in all, there

will be four different Palm grass (Setaria palmifolia) capsule contents.

Capsule Plant Part of the plant used in the

capsule

A Palm grass (Setaria Blade

palmifolia

B Palm grass (Setaria Sheath

palmifolia

C Palm grass (Setaria Root

palmifolia
 D Palm grass (Setaria Seed

palmifolia

C. Treatments and Tests

1. Prothrombin Time (PT) Test on Respondents

Six mice will be collected by the researchers to be used as respondents of the

Palm grass (Setaria palmifolia) capsules..A mouse will be treated with warfarin (?) by

intaking it. A day after, a Prothrombin Time (PT) Test will be conducted on the mouse by

a veterinarian or a professional where a blood sample is taken and the INR results will be

collected to determine the effectiveness of the anticoagulant warfarin. A week after, the

same mouse will be treated with the Palm grass (Setaria palmifolia) blade capsule. A day

after, the process of testing the Prothrombin Time (PT) will be repeated once.

The same process will be repeated for each type of capsule, whether the Palm

grass (Setaria palmifolia) sheath, root or seed capsule. Then, The whole treatment

process will be done in 3 trials to achieve comparable results.

Mouse Trial INR values with INR values with Palm
warfarin treatment grass (Setaria
palmifolia) blade
capsule treatment

A 1

Mouse Trial INR values with INR values with Palm

warfarin treatment grass (Setaria
palmifolia) sheath
capsule treatment

B 1

Mouse Trial INR values with INR values with Palm

warfarin treatment grass (Setaria
palmifolia) root
capsule treatment

C 1

3
Mouse Trial INR values with INR values with Palm
warfarin treatment grass (Setaria
palmifolia) seed
capsule treatment

D 1