CHAPTER 1

PROBLEM: RATIONALE AND ITS BACKGROUND

This chapter contains the background of the study, statement of the

problem, hypothesis of the study, conceptual framework, significance of

the study, scope and delimitations, and the definition of terms.

Rationale

Potassium is an essential mineral micronutrient and is the main

intracellular ion for all types of cells, while having a major role in

maintenance of fluid and electrolyte balance. Potassium is necessary for

the function of all living cells, and is thus present in all plant and animal

tissues. It is essential for proper blood flow, thus, it is needed to prevent

diseases or blood circulation failures. Potassium is the third most abundant

element in human body.

The researchers are aiming to determine the potential of Eleusine

indica in increasing the potassium level in our body. Considering the health

and economic usage of the country, the researchers’ study will renown to

its benefits in health supplies through today in the 21st century.

1|Page
 Potassium is indeed a significant element in our body. Lack of

potassium may cause abnormalities in our body. According to Healthline.

com people who are lack of potassium may experience digestive problem,

heart palpitation, breathing difficulties etc. According also to Mayoclinic.

org, low potassium level is life- threatening and needs urgent medical

attention.

2|Page
 A. Background of the Study

Goose Grass (Eleusine indica) or Paragis for locals, is a species of

grass in the family Poaceae. It is a small annual grass distributed

throughout the warmer areas of the world to about 50o latitude. It is a

serious weed in at least 42 countries. It is an invasive species in some

areas. It thrives in distributed areas with compacted soils in full sun. It

performs C3 photosynthesis and therefore can grow in hot climates and in

the hotter months of the temperate zone.

Studies are conducted to see the potential of Goose Grass in having

antioxidant, antibacterial and anti- cancer effects in our body. This plant

has been the component of “basic remedy” in Vietnamese traditional

medicine and also used for kidney problems in Trinidad and Tobago. Its

roots are depurative, diuretic, febrifuge and laxative and hence are used

for the treatment of influenza, hypertension, urine retention and oliguria.

Hypokalemia is a low- level potassium (K) in the blood serum.

Hypokalemia is one of the most common water- electrolyte imbalances. It

affects about 20% of people admitted to hospital. The word “hypokalemia”

is from hypo- means “under”; kalium meaning potassium, and -emia

means “condition of the blood”. Mildly low- levels do not typically cause
3|Page
symptoms. Symptoms may include feeling tired, leg cramps, weakness and

constipation increases the risk of an abnormal heart rhythm, which is often

too slow and can cause cardiac arrest.

Hypokalemia is rarely caused by consuming too little potassium

because many foods (such as beans, dark leafy greens, potatoes, fish,

and bananas) contain potassium.

Potassium is needed for the contraction of muscles (as well as the

heart muscles) and is needed for the functioning of many complicated

proteins (enzymes). Potassium is found primarily in the skeletal muscles

and bone, and participates with sodium to contribute to the normal blood

flow of body fluids between cells in the body. Roughly 98% of potassium

in your body is found in your cells. Of this, 80% is found in your muscle

cells. While 20% is found in your bones, red blood cells and liver. When

the kidneys are functioning normally, the amount of potassium in the diet

is sufficient.

Hypokalemia was more common in patients 65 years or older than

in younger groups. Women are in more risk than in men. But still younger

women and men are still in risk of having hypokalemia.

4|Page
 High risks of arrhythmia if potassium level drops low. These

patients are also quite likely to be on diuretics and digoxin. Monitor

potassium levels regularly. Potassium levels should also be closely

monitored in diabetic patients due to the adverse effects of insulin and

dextrose on potassium levels and the risk of metabolic acidosis. Patients

with hypertension aims to keep potassium levels at around 4.0 mmol/L,

even a small drop in potassium level should be treated. Any patients on

this combination require close monitoring in order to diagnose a potential

hyperkalemic (high potassium level in blood) as soon as possible.

The researchers came up to the idea in making a product to help

people who are suffering hypokalemia that is affordable, efficient and very

easy to use. The researchers’ study may also help people lower down the

risk of abnormal heart beat, muscles being paralyzed, breathing difficulties

and other serious conditions.

5|Page
 B. Statement of the Problem

This study aims to determine the effectiveness of Paragis extract as

Anti Hypokalemia on Sprague Dawley Rats.

1. Can Eleusine indica increase the potassium level of rats?

2. Which has higher potassium rate, the rats that are given Eleusine indica

extract or the rats that are given contaminated water?

3. Is Eleusine indica extract more effective in augmenting potassium level

of rats rather than K- Durules and Normal Saline Solution?

6|Page
 C. Hypotheses of the Study

Hypotheses 1.

Ho : The Eleusine indica extract CANNOT affect and increase the Potassium

Level of rats.

Ha : The Eleusine indica extract CAN affect and increase the Potassium Level of

rats.

Hypotheses 2.

Ho : The potassium rate of rats that are given Eleusine indica extract is NOT

greater than the rats that are given contaminated water.

Ha : The potassium rate of rats that are given Eleusine indica extract IS greater

than the rats that are given contaminated water.

Hypotheses 3.

Ho : Eleusine indica extract is LESS effective in augmenting potassium level of

rats rather than K- Durules and Normal Saline Solution.

Ha : Eleusine indica extract is MORE effective in augmenting potassium level of

rats rather than K- Durules and Normal Saline Solution.

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 D. Conceptual Framework

The Conceptual Framework which shows the Materials used,

Processes and the Product made during the study.

INPUT PROCESS

◐ Gather Materials
◐ Eleusine
indica ◐ Clean the material and equipment
◐ Potassium properly.
sorbate
◐ Chop the Paragis into small pieces.
◐ Heat it with some water for 10 min.
and reflux. Afterwards, boil it again
and add some potassium sorbate to
OUTPUT
elongate its life.
◐Eleusine indica ◐ Put the extract in a clean vial.
Extract as Anti-
hypokalemia on
Sprague Dawley
Rats.

Figure 1.1

Shows the Conceptual Framework of the Study

8|Page
 E. Significance of the Study

After successively finishing this study, the researchers are expecting

this to be of great contribution to both science and medicine. Likewise, this

may strengthen the worth of using plants as a source of solving health-

related problems.

Hence, bioactive compounds from plants and natural products

provide interest to chemist, biologist, clinicians, and students for

continuous search for more effective, less cost, and less toxic drugs.

This study would also be a great contribution especially to the poor

who can’t afford buying expensive commercial drugs. Hereafter, it is more

convenient if the plant were discovered to significantly prevent or even

treat diseases in a cheaper cost, with the plant already available just

around the community.

Patients suffering from hypokalemia and related diseases can benefit

from the Eleusine indica extract in augmenting their potassium level.

Likewise, it can also be used to lower the risk of cardiac arrest to reduce

the cost of their therapy. As to the students, the results of this study will

serve as a basis of discussion on plants found in the Philippines.

9|Page
 Availability of the resources can also benefit the society, community

and families who have difficulty in accessing commercial drugs. The

awareness of natural products could potentially treat hypokalemia likewise

decrease the expenditure and effort in the procurement of drugs.

Henceforth, this study does not only aim to be useful in the

Philippines, but rather aims to be of great contribution to the neighboring

countries or throughout the world to help solve hypokalemic related

problems just by using the God given resources.

10 | P a g e
 F. Scope and Delimitation

The research paper aims to know the hypokalemic property of

Eleusine indica. The main material used is the Eleusine indica.

Eleusine indica is delimited in only 42 tropical countries. These 42

countries are very fortunate in having this plant because of its helpful

effects.

The researchers started at April 2019 and ended at September 2019.

We began to collect materials, prepare the Eleusine indica chopping it into

small pieces, then, boiling it to extract the juice. Testing at least 24

subjects being certified by Dr. Ike Manuel G. Flores, pathologist and

Marilou M. Sison, medical technologist.

The study was delimited on Eleusine indica as a potassium booster

on rats.

11 | P a g e
G. Definition of Terms:

a. Arrhythmia-is a problem with the rate or rhythm of your

heartbeat.

b. Depurative- herbs that are considered to have purifying and

detoxifying effects. Herbs that are considered depurative include

Lingonberry, Ukshi, the four- leaf clover, Paris polyphylla, and some

species of Elderberry.

c. Digoxin- A poisonous cardio tonic steroid C41H64O14 obtained from a

foxglove and used specially to treat atrial fibrillation.

d. Diuretic- causing increased passing of urine.

e. Electrocardiogram-is a test that checks how your heart is

functioning by measuring the electrical activity of the heart.

f. Electrolyte- any of one various substances in the fluid of your body

that control how your body processes waste and absorbs vitamins,

minerals, etc.

g. Febrifuge- An agent that reduces fever.

h. Goose Grass-is a species of grass in the family Poaceae. It is a

small annual grass distributed throughout the warmer areas of the

world to about 500 latitude.

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i. Hyperaldosteronism-is a disease in which the adrenal gland(s)

make too much aldosterone which leads to hypertension (high blood

pressure) and low blood potassium levels.

j. Hypokalemia- less-than-normal amounts of potassium in the

blood. Signs and symptoms include palpitation, muscle weakness or

cramping, paresthesia, GI complaints such as constipation, nausea

or vomiting and abdominal cramping. Patient may also experience

frequent urination, delirium and depression.

k. Hypomagnesemia- less-than-normal amounts of magnesium in

the blood. Signs and symptoms include nausea, and vomiting,

muscle weakness, tremors, tetany, and lethargy.

l. Hypotheses- a supposition or proposed explanation made on the

basis of limited evidence as a starting point for further investigation.

m. Laxative -are substances that loosen the stools and increase bowel

movement.

n. Micronutrient- a chemical element or substance that is essential in

minute amounts to the growth and health of a living organism.

o. Oliguria-defined as a urine output that is less than 1mL/kg/h in

infants, less than 0.5mL/kg/h in children, and less than 400 mL or

500 mL per 24h in adults- these equals 17 or 21 mL/hour.

13 | P a g e
p. Potassium-the metallic base of potash. Its chemical symbol is K.

q. Potassium Sorbate- white salt that is very soluble in water. It is

used in preserving food.

r. Sprague Dawley Rats- is an outbred multipurpose breed of albino

rats used extensively in medical and nutritional research.

14 | P a g e
 CHAPTER 2

REVIEW OF RELATED LITERATURE

This chapter shows the related literature, related studies and other

supporting details about our research. These details helped us in doing

and supporting our research.

A. Related Legal Basis

Republic Act No. 9502 an act providing for cheaper and quality

medicines.

Section 1. This act shall be known as the “Universally Accessible

Cheaper and Quality Medicines Act of 2008”.

Section 2. Declaration of Policy. – It is the policy of the State to

protect public health and, when the public interest or circumstances of

extreme urgency so require, it shall adopt appropriate measures to

promote and ensure access to affordable quality drugs and medicines for

all.

Pursuant to the attainment of this general policy, an effective

competition policy in the supply and demand of quality and affordable

15 | P a g e
drugs and medicines is recognized by the State as a primary instrument.

In the event that full competition is not effective, the State recognizes as a

reserve instrument the regulation of prices of drugs and medicines, with

clear accountability by the implementing authority as mandated in this Act,

as one of the means to also promote and ensure access to quality

affordable medicines.

Section 3. Construction in Favor of Protection of Public Health. –

All doubts in the implementation and interpretation of the provisions of this

Act, including its implementing rules and regulations, shall be resolved in

favor of protecting public health.

Discoveries, scientific theories and mathematical methods, and in

the case of drugs and medicines, the mere discovery of a new form or new

property of a known substance which does not result in the enhancement

of the known efficacy of that substance, or the mere discovery of any new

property or new use of a known substance, or the mere use of a known

process unless such known process results in a new product that employs

at least one new reactant.

16 | P a g e
Mandate:

Administrative Order 36, s. 2010

Micronutrient deficiencies can cause inter-generational

consequences. The level of health care and nutrition that women receive

before and during pregnancy, at childbirth and immediately post-partum

has significant bearing on the survival, growth and development of their

fetus and newborn. Undernourished babies tend to grow into

undernourished adolescents. When undernourished adolescents become

pregnant, they in turn, may give birth to low-birth weight infants with

greater risk of multiple micronutrient deficiencies.

Micronutrient deficiencies have considerable impact on economic

productivity, growth and national development. Widespread iron deficiency

is estimated to decrease the gross domestic product (GDP) by as much as

2% per year in the worst affected countries. Conservatively, this translates

into a loss of about Php 172 per capita or 0.9% of GDP. Productivity losses

for anemic manual laborers have been documented to be as high as 9%

for severely stunted workers and 5% and 17% for workers engaged in

moderate and heavy physical labor respectively (Micronutrient

Supplementation Manual of Operations)

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Aquino Health Agenda (AHA): Achieving Universal Health Care for All

Filipinos– Kalusugan Pangkalahatan

Its objectives are:

1. Contribute to the reduction of disparities related to nutrition

through a focus on population groups and areas highly affected or at risk

to malnutrition

2. To provide vitamin A capsules, iron and iodine supplements to treat

or prevent specific micronutrient deficiencies

3. Go to scale with key interventions on micronutrient

supplementation, food fortification, salt iodization and nutrient education.

4. Revive, identify, document and adopt good practices and models for

nutrition improvement.

5. Build Nutrition human resource in relevant departments/ agencies.

18 | P a g e
Intervention/ Strategies Employed or Implemented

1. Micronutrient Supplementation- is the provision of

pharmaceutically prepared vitamins and minerals for treatment or

prevention of specific micronutrient deficiency.

2. Food Fortification- the addition of essential micronutrients to widely

consumed food product at levels above its normal state.

3. Improving diet/ dietary diversification- the adoption of proper food

and nutrition practices thru nutrition education food production and

consumption.

4. Growth monitoring and promotion- is an educational strategy for

promoting child health, human development and quality of life

through sequential measurement of physical growth and

development of individuals in the community.

19 | P a g e
 B. Related Literature

Hypokalemia

Hypokalemia is a metabolic imbalance characterized by extremely

low potassium levels in the blood. Potassium is needed for the contraction

of muscles (as well as the heart muscles) and is needed for the functioning

of many complicated proteins (enzymes). Body chemicals and hormones

such as aldosterone also help in regulating potassium in the body.

Hypokalemia occurs, there is an imbalance resulting from a dysfunction in

this normal process, or the rapid loss of urine or sweat without

replacement of sufficient potassium.

Causes

Hypokalemia always occurs as a result of excessive loss of

potassium through the urine, sweat or stool. It is always a symptom of

another disorder rather than a disease that occurs by itself. Causes of

hypokalemia include vomiting, diarrhea, medications like furosemide and

steroids, dialysis, diabetes insipidus, hyperaldosteronism, hypomagnesemia

and not enough intake in the diet. In many adrenal disorders, such

as Cushing syndrome, the adrenal glands produce too much aldosterone,

20 | P a g e
a hormone that causes the kidneys to excrete large amounts of

potassium. The excessive excretion of potassium in the urine (kaliuresis)

may result from the use of diuretic drugs. Hypokalemia may affect both

males and females. However, it occurs more commonly on females.

Signs and Symptoms

Most often hypokalemia has no obvious signs of the disorder.

However, symptoms of hypokalemia may include attacks of severe muscle

weakness, eventually lead to paralysis and possibly respiratory failure.

Severe hypokalemia my also lead to disruption of skeletal muscle cells,

particularly during exercise. In potassium depleted muscle, the lack of

potassium prevents adequate widening of blood vessels, resulting in

decreased muscle blood flow, cramps and destruction of skeletal muscles.

Treatments

The speed at which potassium should be replaced depends on whether

or not there are symptoms or abnormalities on an electrocardiogram.

Potassium levels that are only slightly below the normal range can be

manage with changes in the diet. Lower levels of potassium require

replacement with supplements either taken by mouth or given

21 | P a g e
intravenously. If given intravenously, potassium is generally replaced at

rates of less than 20mmol/hour. Solutions containing high concentrations

of potassium (>40mmol/L) should be generally be given using a central

venous catheter.

22 | P a g e
 C. Related Studies

Plantae

Angiosperms

Poales

Poaceae

Eleusine

E.indica

Goose Grass (Eleusine indica)

Laboratory of Cancer Research MAKNA-UPM, Institute of Biosciences

(IBS), University Putra Malaysia, Serdang, Selangor, Malaysia Department

of Biochemistry and Molecular Biology, Faculty of Medicine and Health

Sciences, University of Sana’a, Sana’a, Yemen Department of Biomedical

Sciences, Faculty of Medicine & Health Sciences. The use of evidence-

based complementary and alternative medicine is increasing rapidly.

Eleucine indica (EI) is traditionally used in ailments associated with liver

and kidneys. The therapeutic benefit of the medicinal plants is often

attributed to their antioxidant properties. Therefore, the aim of this study

was to screen the hexane, dichloromethane, ethyl acetate (EA) and

methanol extracts (MeTH) of EI for their antioxidant, antibacterial and

23 | P a g e
anti-cancer effect using total phenolic contents (TPCs) and DPPH. Ash of

leaves contain SiO 16- 47%; CaO 10- 13%; and chlorine, 6-7%. Study

showed the dry matter content to be 35.8%, crude protein 12.4%.

Methanol extract yielded flavonoids, tannins, alkaloids, cardiac glycosides,

anthraquinones. Elemental analysis yielded: potassium, calcium,

magnesium, phosphorus, copper, iron, manganese, molybdenum, and

zinc. Calorific value was 3753 ± 223. In a study of ten botanicals, E. indica

yielded the highest calcium content (165.0 mg/100g). Iron content was

9.7 mg/100g. the plant also yielded alkaloids, carotenoids, flavonoids,

saponins, and steroids.

Total Phenolic Compounds (TPC) of the EI extracts was determined

using Folin- Ciocalteu reagent method. Briefly, stock solutions of EI

extracts were prepared in a concentration of 20? Mg/ml. Fifty microliters of

this solution were transferred to test tubes (n = 3). To this, 0.4 of Folin-

Ciocalteu reagent (1:10) was added and mixed thoroughly. After 1 minute,

0.8ml of sodium bicarbonate solution (NaHCO3 7.55%) was added and the

mixture was allowed to stand for 30 minutes with intermittent shaking.

Absorbance was measured at 765 nm using a Shimadzu UV- ViS

spectrophotometer. The TPC was expressed as gallic acid equivalents

24 | P a g e
(GAE) in mg/g extract, obtained from the standard curve of Gallic

solutions.

The reaction involves DPPH reacting with an antioxidant compound,

the latter donating hydrogen and reduced, which then leads to the change

of DPPH.

The antimicrobial activity of plant extracts was evaluated using two

Gram-positive bacteria, methicillin resistant Staphylococcus aureus (MRSA)

and Bacillus subtilis B29, and other two Gram-negative bacteria,

Pseudomonas aeruginosa 60690 and Salmonella choleraesuis. All the

bacterial stains were obtained from the Laboratory of Molecular

Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, Serdang,

Malaysia.

MCF- 7 human breast cancer cells and HT- 29 human colon carcinoma

cells were purchased from the American Type Culture Collection (ATCC),

USA. Human T4-lymphoblastoid cell line, CEM-SS, used in this study were

obtained from The NIH AIDS Research and References Reagent Program,

USA. The cancer cells were grown at 37℃ in humidified CO2 incubator with

5% CO2 in RPMI- 1640 media supplemented with 10% fetal bovine serum

(Invitrogen Corp., Auckland, New Zealand).

25 | P a g e
 The microbial effects of EI extracts studied are summarized. The

antibacterial activities of both different extracts of EI were evaluated using

both Gram-positive and Gram-negative bacteria. The highest antibacterial

activity observe was obtained by hexane extract on MRSA, while the DCM

extract showed weak activity on P. aeruginosa. Showed a broad-spectrum

activity, compared with the positive control (streptomycin) against all

tested bacteria except for B. subtilis, which showed resistance to all

extracts of EI. In contrast, MeTH did not exhibit any antibacterial activity

towards MRSA, B. subtilis B29, P aeruginosa 60690 and S. choleraesuis.

The positive control, streptomycin, had shown zones of inhibition of 20 ±

1.5, 20 ± 1.3, and 23 ± 1.0mm in MRSA, P. aeruginosa, S choleraesuis,

and B subtilis, respectively. Interest in finding naturally occurring

antioxidants for use in foods or medicinal materials to replace synthetic

antioxidants has been increased considerably. Restrictions on the use of

synthetic antioxidants such as butylated hydroxyanisole (BHA) and BHT

are being imposed due to their carcinogenicity. Therefore, a need for

identifying alternative natural and safe sources of antioxidants, especially

of plant origin, has increased in recent years. In this study, the hexane,

DCM, EA and MeTH of EI were assayed for their antioxidant, antibacterial

and anti- cancer properties using TPCs and DPPH, disc diffusion method
26 | P a g e
and MTT cytotoxicity assay test. The extraction procedure used was cold

maceration, in which the solvents used were in the order of increasing

polarity.

The therapeutic benefit of the medicinal plants is usually contributed

to their antioxidant properties. Phenolic compound possesses diverse

biological activities such as anti- inflammatory, anti- carcinogenic and anti-

atherosclerotic activities. These activities might be related to their

antioxidant activity. Other studies showed that there were significant

correlations between phenolic compounds and antioxidant properties of

medicinal plants under investigations. In this respect, a part of our study

was determining TPC in different extracts obtained from EI.

Potassium Sorbate

Potassium sorbate is the potassium salt of sorbic acid, with a

chemical formula of CH3- CH= CH- CH= CH- CO2K. It is a white salt- like

that is very soluble in water. It is primarily used as a food preservative.

27 | P a g e
 D. Justification of the Study

The study aimed to find possible prevention and cure to hypokalemia

which can help the community. Despite those limitations of the product, it

can still help many people, specially to those who have hypokalemia.

Different studies shown us that Eleusine indica have many positive effects

in our body.

28 | P a g e
 CHAPTER 3

MATERIALS AND METHODS

This chapter indicates the information on the different methods and

procedures done in this study. This chapter also indicates the preliminary

screening and biological assay method which was employed in Eleusine

indica. The experimental design used by the researchers is the CRD

method.

Research Design

Experimental method of research is the use of controlled

observations and measurements in testing the hypotheses. This was used

by the researchers in identifying the potassium rate of Eleusine indica. This

method of investigatory project was utilized to obtain results of

phytochemical testing and screening and biological assay which were

recorded and interpreted. The researchers used CRD (Completely

Randomized Design) as a research design.

Albino rats specifically Sprague Dawley Rats weighting 150- 200

grams were used as test animals. Rats are being used as the test animals

29 | P a g e
because it is not ethically acceptable to use humans without further

studies on animals.

ALBINO RAT TREATMENT RANK

903 A 10
736 A 4
204 A 15
192 A 5
281 A 1
500 B 13
034 B 6
486 B 14
830 B 9
976 B 2
029 C 7
209 C 3
836 C 13
205 C 8
412 C 11
Table 3.1

Shows the CRD of the research

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Locale of the Study

Goose grass (Eleusine indica) or Paragis were gathered from

Camagsingalan, Sual, Pangasinan. All the Goose Grass were gathered from

a single area for uniformity purposes. On the other hand, the test animals

were procured from an accredited supplier in Barangay Agtas, San Carlos

City, Pangasinan. In pursuant to the provision of Republic Act 8485 or the

Animal Welfare Act of 1988 and DA- Administrative Order (AO) No. 40

series of 1999, on the Rules and Regulations on the Scientific Procedure

Using Animals.

Biological Assay and potassium reading were undertaken in AGS

Laboratory, AUJAN Medics. Meanwhile, the blood collection of the rats was

undertaken in the same laboratory under the supervision of Mr. Alvin C.

Franza, a graduate of Bachelor of Science in Medical Laboratory Sciences.

Corollary, potassium count reading was conducted in the AGS Laboratory,

AUJAN Medics in which laboratory personnel assisted us.

31 | P a g e
Research Animals

Animal Procurement

Twenty- Four Albino Rats weighting 150- 200 grams were used as

the subjects of the blood potassium reading. Albino rats were purchased

on an accredited breeder in San Juan Agtas, San Carlos City, Pangasinan.

Ethical Considerations

Improvement of human health is the main goal of the study.

However, it is unethical to use humans without first conducting studies on

animals to ensure the safety and efficiency of the procedure or drug

products.

Animal Disposal

At the end of the assay procedure, any animal left alive was

painlessly killed immediately and disposed of properly, preferably by

incineration.

Tail- tipping Method

Prior to collection of blood sample, the tail of the rats was cleansed

using wet cotton previously dipped in warm water. The tail of the rats was

32 | P a g e
shaved where its vein is prevalent and lacerated by enough depth to

rupture its vein. Blood samples are collected using non- anticoagulant

tubes (Red Top) and will be ready for analysis.

Proper Storage of the Medicine

There are several things to consider when storing medicines. The

first one to consider is the temperature. Heat and moisture may affect the

medicine. Make sure that the medicine is placed in a cool and dry place

not exceeding 300C. Second, always keep the medicine in its original

container. Lastly, always store it in a place that is out of reach and sight of

children.

33 | P a g e
GENERAL PROCEDURE

Preparation of Aqueous Extract

Prepare plant sample, rinse and cut into smaller pieces. Place at

least 100 grams of the sample into a 500ml Erlenmeyer’s flask. Pour 300

mL of Distilled water measured using graduated cylinder and seal the

flasks opening with cork. Heat it for one hour and then reflux for another

hour. Cool down and filter the extract, using filter paper and funnel, to

another Erlenmeyer’s flask. Add 60mL of potassium sorbate to prolong its

life. The ratio of the extract to the potassium sorbate should be 5:1. Stir

thoroughly. The aqueous extract is now ready.

With these given methods, you can make at least five 50mL of

Aqueous Extract that can be consumed in a week.

Administration of Contaminated Water

Before the experimentation, the rats were weighted to determine its

initial weight. The blood is collected using tail- tipping method to

determine its potassium rate. After getting its baseline, the rats were

administered with a contaminated water from a fishpond. This will reduce

34 | P a g e
its baseline. The amount of contaminated water was measured carefully

using this formula- ml/L= (initial weight× 3mL) ÷ 1000.

ALBINO RAT INITIAL WEIGHT POTASSIUM COUNT

NUMBER (in grams) (in mEq/L)
1 200 5.6
2 200 6.2
3 200 5.1
4 191 6.0
5 193 6.7
6 179 5.3
7 189 6.2
8 195 5.2
9 182 5.8
10 170 6.3
11 184 6.1
12 199 6.3
13 190 5.3
14 195 6.0
15 200 5.5
16 180 5.8
17 197 6.1
18 174 5.8
19 190 6.9
20 197 5.6
21 172 5.3
22 163 6.2
23 185 5.5
24 188 5.8
Table 3.2

Shows the initial weight and initial potassium count of test animals.
35 | P a g e
 ALBINO RAT COMPUTATION
NUMBER (initial weight× 3mL) ÷ 1000
1 (200× 3mL) ÷ 1000= 0.60
2 (200× 3mL) ÷ 1000= 0.60
3 (200× 3mL) ÷ 1000= 0.60
4 (191× 3mL) ÷ 1000= 0.57
5 (193× 3mL) ÷ 1000= 0.58
6 (179× 3mL) ÷ 1000= 0.54
7 (189× 3mL) ÷ 1000= 0.57
8 (195× 3mL) ÷ 1000= 0.59
9 (182× 3mL) ÷ 1000= 0.55
10 (170× 3mL) ÷ 1000= 0.51
11 (184× 3mL) ÷ 1000= 0.55
12 (199× 3mL) ÷ 1000= 0.60
13 (190× 3mL) ÷ 1000= 0.57
14 (195× 3mL) ÷ 1000= 0.59
15 (200× 3mL) ÷ 1000= 0.60
16 (180× 3mL) ÷ 1000= 0.54
17 (197× 3mL) ÷ 1000= 0.59
18 (174× 3mL) ÷ 1000= 0.52
19 (190× 3mL) ÷ 1000= 0.57
20 (197× 3mL) ÷ 1000= 0.59
21 (172× 3mL) ÷ 1000= 0.52
22 (163× 3mL) ÷ 1000= 0.49
23 (185× 3mL) ÷ 1000= 0.56
24 (188× 3mL) ÷ 1000= 0.56
Table 3.3

Shows the computation of the measurement of the contaminated

water

36 | P a g e
ALBINO RAT ADMINISTRATION POTASSIUM COUNT
NUMBER (in mL) (in mEq/L)
1 0.6 5.0
2 0.6 5.7
3 0.6 4.6
4 0.57 5.3
5 0.58 6.2
6 0.54 4.6
7 0.57 5.7
8 0.59 4.5
9 0.55 5.3
10 0.51 5.5
11 0.55 5.6
12 0.6 5.8
13 0.57 4.6
14 0.59 5.8
15 0.6 5.0
16 0.54 5.1
17 0.59 5.5
18 0.52 5.3
19 0.57 6.2
20 0.59 5.0
21 0.52 4.8
22 0.49 5.6
23 0.56 5.1
24 0.56 5.2

Table 3.4

Shows the result after the administration of contaminated water

37 | P a g e
Administration of Plant Extract

After the administration of contaminated water from fishponds, the

potassium count of the rats dropped. After 7 days the researchers

administered 95% concentration of the plant aqueous extract. Laboratory

personnel assisted the researchers in administration of plant extract.

ALBINO RAT FINAL WEIGHT POTASSIUM COUNT

NUMBER (in grams) (in mEq/L)
1 205 5.5
2 204 6.0
3 205 5.3
4 196 6.2
5 197 6.4
6 182 5.0
7 194 5.8
8 199 4.9

Table 3.5

Shows the results after administration of Eleusine indica aqueous extract

38 | P a g e
Administration of K- durules

Kalium Durules (K- Durules) is a potassium supplements are

indicated in patients with hypokalemia, with or without metabolic alkalosis;

in chronic digitalis intoxication; and in patients with hypokalemic familial

periodic paralysis. The researchers administered k- durules to show how

the researchers’ product can go with commercially produce potassium

supplements like this one.

The researchers considered the following reminders in

Administrating Kalium Durules to albino rats.

1. The researchers administered each dose with meals mixed in water

and other suitable liquid.

2. The researchers administered the medicine following the frequency

and amount according to the rat’s mass.

3. The researchers administered the medicine through oral ingestion by

using syringe.

4. The researchers also checked if the rats show evidence of

gastrointestinal bleeding.

39 | P a g e
 ALBINO RAT FINAL WEIGTH POTASSIUM COUNT
NUMBER (in grams) (in mEq/L)
9 186 6.6
10 174 6.9
11 188 6.5
12 203 6.8
13 195 5.5
14 200 6.8
15 105 6.1
16 184 6.4

Table 3.6

Shows the results after administration of k- durules (positive control)

Administration of Normal Saline Solution

Sodium and potassium are important electrolytes needed for the

body to function normally with the help maintain fluid and blood volume in

the body. However, a person can get high blood pressure by consuming to

much sodium and not enough potassium. The following table shows how

sodium can affect rats’ potassium level.

40 | P a g e
 ALBINO RAT FINAL WEIGHT POTASSIUM COUNT
NUMBER (in grams) (in mEq/L)
17 201 5.6
18 177 5.5
19 194 6.0
20 201 5.1
21 176 4.9
22 167 5.8
23 189 5.0
24 192 5.4

Table 3.7

Shows the results after administration of saline solution (negative control)

Tools for Data Analysis

The results of the phytochemical screening were recorded, noted

and compared from standard positive results to determine the active

constituents present in Eleusine indica which is responsible for augmenting

the potassium count. Likewise, the potassium count of the rats treated

with Eleusine indica were computed to determine the average potassium

41 | P a g e
augmenting activity of the plant samples individually. On the other hand, t-

test was used to determine if there is significant difference among the

group of rats treated with Eleusine indica before and after treatment based

on potassium count and f- test (ANOVA) to determine if there is significant

difference among rats that are administered with Paragis Aqueous Extract,

Kalium Durules (K- durules) and normal saline solution. Moreover, the

results were interpreted clinically.

42 | P a g e
 Gathering and Collecting the Raw Materials

Gathering Materials

Extracting Raw Materials

Testing the Effectiveness of the Product

Analysis of Data

Constructing and Finalizing the Research

Output

Figure 3.1

Shows the Systematized Process of Research

43 | P a g e
 CHAPTER 4

RESULTS AND DISCUSSIONS

This chapter provides the results and discussions of the study. Data

gathered, generate and collected were examined through Statistical

Analysis. This chapter also provides the positive and negative controls

performed in the study. This were all summarized and presented below

using graphs for easy comprehension. Discussions are likewise presented

below.

Note:

Group A- Rats 1- 8 (Normal Control Group)

Group B- Rats 9- 16 (Positive Control Group)

Group C- Rats 17- 24 (Negative Control Group)

44 | P a g e
 Y Potassium Baseline of Albino Rats

7 6.7
6.2 6.2
6
6 5.6
5.3 5.2
5.1
5

0
Rat 1 Rat 2 Rat 3 Rat 4 Rat 5 Rat 6 Rat 7 Rat 8 X

Figure 4.1

Shows the graph of Group A’s Potassium Baseline. This may serve as the

initial data of Group A (rats 1-8) or the normal control group.

45 | P a g e
 Y Potassium Baseline of Albino Rats

7
6.3 6.3
6.1 6
5.8
6 5.5
5.3 5.2

0
Rat 9 Rat 10 Rat 11 Rat 12 Rat 13 Rat 14 Rat 15 Rat 16 X

Figure 4.2

Shows the graph of Group B’s Potassium Baseline. This may serve as the

initial data of Group B (rats 9-16) or the positive control group.

46 | P a g e
 Y Potassium Baseline of Albino Rats

6.9
7
6.1 6.2
5.8 5.8
6 5.6 5.5
5.3

0
Rat 17 Rat 18 Rat 19 Rat 20 Rat 21 Rat 22 Rat 23 Rat 24 X

Figure 4.3

Shows the graph of Group C’s Potassium Baseline. This may serve as the

initial data of Group C (rats 17-24) or the negative control group.

47 | P a g e
Y 2nd Reading: Potassium Level of Rats

7
6.2

6 5.7 5.7
5.3
5
5 4.6 4.6 4.5

0
Rat 1 Rat 2 Rat 3 Rat 4 Rat 5 Rat 6 Rat 7 Rat 8 X

Figure 4.4

Shows the graph of Group A’s Potassium Level, 7 days after the

Administration of Contaminated Water from Fishponds.

48 | P a g e
Y 2nd Reading: Potassium Level of Rats

5.8 5.8
6 5.6
5.5
5.3
5 5.1
5 4.6

0
Rat 9 Rat 10 Rat 11 Rat 12 Rat 13 Rat 14 Rat 15 Rat 16 X

Figure 4. 5

Shows the graph of Group B’s Potassium Level, 7 days after the

administration of contaminated water from fishponds.

49 | P a g e
Y 2nd Reading: Potassium Level of Rats

7
6.2

6 5.5 5.6
5.3 5.2
5 5.1
4.8
5

0
Rat 17 Rat 18 Rat 19 Rat 20 Rat 21 Rat 22 Rat 23 Rat 24 X

Figure 4.6

Shows the graph of Group C’s Potassium Level, 7 days after the

administration of contaminated water from fishponds.

50 | P a g e
 Y 3rd Reading: Potassium Level of Rats

7
6.4
6.2
6
5.8
6 5.5
5.3
5 4.9
5

0
Rat 1 Rat 2 Rat 3 Rat 4 Rat 5 Rat 6 Rat 7 Rat 8 X

Figure 4.7

Shows the graph of Group A’s Potassium, 7 days after Administration of

95%Eleusine indica extract in the form of Aqueous Extract. This may serve

as the final potassium reading of Group A.

51 | P a g e
 Y 3rd Reading: Administration of K- Durules

6.9 6.8 6.8

7 6.6 6.5 6.4
6.1
6 5.5

0
Rat 9 Rat 10 Rat 11 Rat 12 Rat 13 Rat 14 Rat 15 Rat 16 X

Figure 4.8

Shows the graph of Group B’s Potassium, 7 days after Administration of K-

Durules. The datum that are shown above are the results.

52 | P a g e
 3rd Reading: Administration of Saline
Solution
6
Y

5.8
6 5.6 5.5 5.4
5.1 5
4.9
5

0
Rat 17 Rat 18 Rat 19 Rat 20 Rat 21 Rat 22 Rat 23 Rat 24 X

Figure 4.9

Shows the graph of Group C’s Potassium, 7 days after Administration of

Normal Saline Solution.

53 | P a g e
 The t-test was employed in the study to test if there is a significant

difference between the potassium level of rats that are given Eleusine

indica or rats that are given contaminated water. This also indicates

whether the null hypothesis is accepted or rejected.

Degrees of Level of

tcomp ttab Freedom Significance DECISION

Reject H0;

4.77 1.714 14 0.01 Accept Ha

Table 4.1

Decision which tells whether the Null Hypothesis is Accepted or Rejected

(T-Test)

54 | P a g e
 The f-test (ANOVA) was employed in the study to test if there is a

significant difference between the EFFECTIVITY of the variables Eleusine

indica extract, K- Durules and Normal Saline Solution. This also indicates

whether the null hypothesis is accepted or rejected.

Degrees of Level of

Fcomp Ftab Freedom Significance DECISION

DFT= 23 Reject H0;

10.82 5.7805 DFB= 2 0.01 Accept Ha
DFW= 21

Table 4.2

Decision which tells whether the Null Hypothesis is Accepted or Rejected

(ANOVA)

55 | P a g e
 CHAPTER 5

SUMMARY OF FINDINGS, CONCLUSIONS AND

RECOMMENDATIONS

This chapter presents the summary of findings, conclusions, and

recommendations of the researchers. This chapter also provides the

bibliography, appendix, and pictorials of the study.

A. Summary of Findings

In this research project, the researchers formulated a medicine

out of Eleusine indica or locally known as Paragis that is available

everywhere.

The test animals (albino rats) where grouped into three. The

first one is the experimental group (Rats 1- 8), the second one is the

positive control group (Rats 9- 16), and the last one is the negative

control group (Rats 17- 24). Three readings are also conducted in the

study. The first reading was the baseline. The second one is the

administration of contaminated water. And the last one is the

administration of Paragis Aqueous Extract. The researchers used t- test

56 | P a g e
and ANOVA as a statistical tool. The computed value for t- test was

4.77. And 10.82 for ANOVA.

In this study the laboratory test that is conducted in A.G.S

Laboratory, AUJAN Medics proves that Paragis extract is effective in

treating hypokalemia.

57 | P a g e
 B. Conclusions

Based on the data that the researchers’ gathered, Eleusine indica

extract CAN affect and increase the Potassium Level in rats. It is also

observable that the potassium rate of rats that are given Eleusine indica

extract IS greater than the rats that are not given Eleusine indica extract.

Based on the computed value in ANOVA, which is 10.82 and is

greater than the value of FTAB, 5.7805. Therefore, Paragis Aqueous Extract

is more effective in augmenting potassium level of rats.

Thus, all alternative hypotheses are accepted.

58 | P a g e
 C. Recommendations

From the above summary of datum and outcomes, the researchers

propose the Paragis as a potassium booster in our body. It can be also

used by doctors to increase the potassium level of their patients. But the

researchers do not recommend the product as medicine to all kinds of

diseases.

The researchers recommend that if someone wants to try to make

their own Paragis Aqueous Extract, make sure to follow all the instructions

that is provided in CHAPTER 3, and ask for surveillance of an adult.

Lastly, the researchers recommend to the next researcher/s to

continue and develop the study for further refinement of the research.

59 | P a g e
D. Bibliography

TM
 Wolters Kluwer (July 2019), IBM Watson Micromedex (Aug.

2019), Cerner Multum TM

(Aug. 2019), “Kalium Durules”, Drugs.

Com.

 Nall, Rachel, “What to know about potassium deficiency symptoms.”

Medical News Today. MediLexicon, Intl., 29 Apr. 2019.

 Eslit, Nila, “Paragis”, Cebu, Philippines, 25 Jan. 2018.

 Stuart M.D., Godofredo Jr. “Paragis”, StuartXChange, August 2018.

 Lewis III M.D., James, “Hypokalemia Low Level of Potassium in the

Blood)”, Brookwood Baptist Health and Saint Vincent’s Ascension

Health, Birmingham, United Kingdom, Sep. 2018.

 Hecht, M., Choi, MD, Justin, “Everything You Should Know About

Potassium Sorbate”, Healthline. Com, 22 Mar. 2017.

 Rogers M.D., Graham, “Hypokalemia”, Healthline. Com, 31 Oct.

2016

 Al- Zubairi, Adel, et al., Laboratory of Cancer Research MAKNA-

UPM, Institute of Biosciences (IBS), Universiti Putra Malaysia,

Serdang, Selangor, Malaysia, 25 June 2009.

 Fujihara, Hideyoshi, et al., Departments of Molecular Physiology and

Biological Physics, Pathology and Internal Medicine, University of

60 | P a g e
 Virginia Health Sciences Center, Charlottesville, Virginia, USA., 19

Sep. 1997.

 Kleinfeld M, et al., “Hypokalemia: are elderly females more

vulnerable”, J Natl Med Assoc. Nov. 1993.

61 | P a g e
E. References

 Al- Zubairi, Adel, et al., Retrieved May 26, 2019 from

https://www.hindawi.com/journals/ecam/2011/965370/

 Stuart M.D., Godofredo Jr., Retrieved July 17, 2019 from

https://www.stuartxchange.com/Paragis.html

 Eleusine indica – Wikipedia. Retrieved August 4, 2019 from

https://www.en.m.wikipedia.org

 Fujihara, Hideyoshi, et al., Retrieved May 26, 2019

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC25718

 Eslit, Nila, Retrieved August 31, 2019

https://wsimag.com/wellness/35291-paragis

 Lewis III M.D., James, Retrieved May 29, 2019

https://www.msdmanuals.com/home/hormonal-and-metabolic-

disorders/electrolyte-balance/hypokalemia-low-level-of-potassium-in-

the-blood

TM TM
 Wolters Kluwer , IBM Watson Micromedex, Cerner Multum ,

Retrieved September 1, 2019

https://www.drugs.com/mmx/kalium-durules.html

 Hecht, M., Choi M.D., Justin, Retrieved June 15, 2019

https://www.healthline.com/health/potassium-sorbate#source
62 | P a g e
 Nall, Rachel, Retrieved August 16, 2019 from

https://www.medicalnewstodsy.com/articles/325065.php

 Rogers M. D., Graham, Retrieved August 31, 2019

https://www.healthline.com/health/hypokalemia

63 | P a g e
 APPENDICES

APPENDIX A

2nd
5.0 5.7 4.6 5.3 6.2
READING
3RD
5.5 6.0 5.3 6.2 6.4
READING
RDIFFERENCE 0.5 0.3 0.7 0.9 0.2

4.6 5.7 4.5

5.0 5.8 4.9

0 .4 0.1 0.4 = 3.5

64 | P a g e
 APPENDIX B

d ̅
𝒅 ̅ )2
(d - 𝒅

0.5 0.4375 0.004

0.3 0.4375 0.019

0.7 0.4375 0.069

0.9 0.4375 0.214

0.2 0.4375 0.056

0.4 0.4375 0.001

0.1 0.4375 0.114

0.4 0.4375 0.001

̅ = 0.4375
𝒅 𝜮= 0.48

Degrees of Freedom= 7

65 | P a g e
 APPENDIX C

̅ )2
∑ (x-x 0.48
Sd2= Sd2= √Sd2 = √𝟎. 𝟎𝟕
n-1 7

0.48
Sd2= Sd2= 0.07 Sd= 0.26
8-1

̅ √n
d
t= sd

0. 4375√8
t= 0. 26

(0.4375)(2.83)
t= 0.26

1.24
t= 0.26

t= 4.77

66 | P a g e
 APPENDIX D

X SUM AVE. VARIANCE

A 8 45.1 5.64 0.31

B 8 51.6 6.45 0.21

C 8 43.3 5.41 0.15

𝐱 𝐱̃ (𝐱 − 𝐱̃)𝟐 𝐱 𝐱̃ (𝐱 − 𝐱̃)𝟐

5.5 5.64 0.02 6.6 6.45 0.02

6.0 5.64 0.13 6.9 6.45 0.20

5.3 5.64 0.12 6.5 6.45 0.002

6.2 5.64 0.31 6.8 6.45 0.12

6.4 5.64 0.58 5.5 6.45 0.90

5.0 5.64 0.41 6.8 6.45 0.12

5.8 5.64 0.03 6.1 6.45 0.12

4.9 5.64 0.55 6.4 6.45 0.002

67 | P a g e
 APPENDIX E

𝐱 𝐱̃ (𝐱 − 𝐱̃)𝟐

5.6 5.41 0.04

5.5 5.41 0.008

6.0 5.41 0.35

5.1 5.41 0.10

4.9 5.41 0.26

5.8 5.41 0.15

5.0 5.41 0.17

5.4 5.41 0.001

TSS= 9.47

SSB= 4.76

SSW= 4.71

68 | P a g e
 APPENDIX F

Degrees of Freedom

DFT= n- 1 DFB= k- 1 DFW= DFT- DFB

= 24- 1 = 3- 1 = 23- 2

= 23 =2 = 21

Mean of Sum of Squares

A. Sum of Squares Between Column

ssB
MSSB= MSSB= 2.38
DFB

MSSB= 4.76
2

B. Sum of Squares Within Column

ssW
MSSW= MSSW= 0.22
DFW

MSSW= 4.71
21

69 | P a g e
 APPENDIX G

Tab Value

𝑫𝑭𝑩
FT= FT= 0.10
𝑫𝑭𝒘

FT= 2
21

Value of Statistical Test

𝐌𝐬𝐬𝐁
FC= FC= 10.82
𝐌𝐬𝐬𝐰

FC= 2.38
0.22

70 | P a g e
 APPENDIX H

Pictorials

71 | P a g e
72 | P a g e
 APPENDIX I

Republic of the Philippines

DEPARTMENT OF EDUCATION
Region I
SCHOOLS DIVISION OFFICE I PANGASINAN

SUAL NATIONAL HIGH SCHOOL

Sual

October 7, 2019

CERTIFICATION

This is to certify that ADRIENNE DE VERA, RUSSEL GEN J.

SEVILLENA and SAMANTHA VALERIE ROMASOC are bonafide Grade 9

students of Sual National High School. Their Science Investigatory Project (SIP) entitled

Anti-Hypokalemia Properties of Goose Grass (Eleusine indica) is an official entry in the

2019 Division Science and Technology Fair.

MARIELA N. CALIMA, Ed.D.

Principal IV

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