Utilizing Moringa Seeds (Moringa oleifera), Jute Leaves

(Corchorus olitorius), and Guava Leaves (Psidium guajava)

for Portable Water Purification: LeafFusion Pouch

A Science Investigatory Project (SIP)

Presented to the Faculty of Senior

High School Department

St. Therese Educational Foundation of Tacloban, Inc.

In Partial Fulfillment of the

Requirements for the Subject

Practical Research I

By

Mary Julianne B. Asid

Jovette Nicole G. Candido

Sef Titus D. Dacoycoy

Euville Niña A. Tabuyan

Cedrick L. Senal

November 2024

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 TABLE OF CONTENTS
TITLE PAGE.............................................................................................................................i

TABLE OF CONTENTS...............................................................................................…...…ii

CHAPTER I.
INTRODUCTION.....................................................................................................................1

Background of the study...................................................................................................….......1

Objective of the Study........................................................................................................…….2

Statement of the Problem............................................................................................................2

Null Hypothesis & Alternative Hypothesis….................................................................…........3

Significance of the Study.............................................................................................................4

Conceptual Framework......................................................................................................….....4

Scope and Limitations.................................................................................................................5

CHAPTER II.
REVIEW OF RELATED LITERATURE...............................................................................6

Related Literature and Studies.................................................................................................…6

CHAPTER III.
METHODOLOGY.....................................................................................................................9

Materials and Equipment ........................................................................................……........…9

Set-up………………………………….............................................................................…......9

Flowchart of the procedure..........................................................................................................10

Experimental Procedure...............................................................................................................10

Data Collection ans Analysis.......................................................................................................14

Timeline.......................................................................................................................................15

REFERENCES...........................................................................................................................16

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 CHAPTER I

INTRODUCTION

Background of the Study

Access to clean and safe water is a significant global challenge, especially in rural and

underserved areas where infrastructure and resources for modern water purification systems are

limited. Contaminated water sources are the major cause of waterborne diseases like diarrhea,

cholera, and typhoid, which are common in communities that have restricted access to affordable

purification methods. This requires innovative, low-cost, and sustainable solutions that are

practical for widespread use in these areas.

Plant-based materials that possess intrinsic coagulating and antimicrobial features are

considered promising candidates for water purification. This research aims to determine the

possibility of employing Moringa seeds (Moringa oleifera), Jute leaves (Corchorus olitorius),

and Guava leaves (Psidium guajava) as sources of natural purifying substances. Moringa seeds

have antibacterial properties and, due to their ability to bind impurities, act as a natural clarifier.

In the case of Jute leaves, they have in-built flocculation properties that help in aggregation,

followed by removal of the suspended particles. Guava leaves possess coagulant properties with

successful results in reducing the turbidity and capturing dangerous microorganisms.

These plant-based materials would then be encapsulated within the portable, teabag-like

innovation that is called the LeafFusion Pouch. Designed to purify water for purposes other than

drinking, including irrigation, sanitation, and various household uses, the LeafFusion Pouch has

compounds that synergistically work upon being placed in contaminated water to bind

contaminants, reduce microbial loads, and improve the clarity and quality of the water.

This research will evaluate the effectiveness of the LeafFusion Pouch in reducing

contamination and enhancing water safety, with an ultimate goal of developing a low-cost,

sustainable, and locally accessible solution for resource-limited communities. In doing so, this

study attempts to contribute to the efforts in mitigating waterborne diseases and improving water

quality in resource-limited settings through the potential of these natural materials.

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 Objectives of the Study

This study focuses on developing a practical solution that utilizes Moringa seeds, Jute

leaves, and Guava leaves in the form of a "LeafFusion Pouch" to effectively purify unfiltered

water. Specifically it aims:

1. To explore if the blend of three natural coagulants with a mixture of variables will

be more effective on lowering the turbidity levels of unfiltered water.

2. To provide a sustainable, low-cost solution of unfiltered water in the rural areas.

3. To discern and recommend the appropriate ratio of Moringa oleifera seeds,

Corchorus olitorius, and Psidium guajava leaves pouch for unfiltered water.

Statement of the Problem

This research aims to investigate the capability of this water filtration pouch, in

developing a portable and affordable water purification system. The following are the main

research questions of the study:

General Problem:

What is the effectiveness of Moringa seeds, jute leaves, and guava leaves

in removing various water contaminants, including pathogens and turbidity, and

how do their purification efficiencies compare under different conditions?

Sub-Problem:

1. What is the optimal ratio of Moringa oleifera seeds, Corchorus olitorius leaves,

and Psidium guajava leaves in a composite pouch for effective purification of

unfiltered water?

i. 1:1 coagulant ratio

ii. 1:2 coagulant ratio

iii. 1:3 coagulant ratio

iv. 2:1 coagulant ratio

v. 3:1 coagulant ratio

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 2. How do variations in the ratio of these materials affect key water quality

parameters, such as contaminant removal efficiency and turbidity reduction?

i. Biochemical Oxygen Demand (BOD)

i. 1:1 coagulant ratio

ii. 1:2 coagulant ratio

iii. 1:3 coagulant ratio

iv. 2:1 coagulant ratio

v. 3:1 coagulant ratio

ii. Chemical Oxygen Demand (COD)

i. 1:1 coagulant ratio

ii. 1:2 coagulant ratio

iii. 1:3 coagulant ratio

iv. 2:1 coagulant ratio

v. 3:1 coagulant ratio

Null Hypothesis

The LeafFusion Pouch, composed of Moringa seeds (Moringa oleifera),

jute leaves (Corchorus olitorius), and guava leaves (Psidium guajava), does not

significantly improve water quality parameters (e.g., turbidity, microbial load, and

chemical contaminants) compared to untreated water.

Alternative Hypothesis

The LeafFusion Pouch, composed of Moringa seeds (Moringa oleifera),

jute leaves (Corchorus olitorius), and guava leaves (Psidium guajava), significantly

enhances water quality by reducing turbidity, lowering microbial load, and minimizing

chemical contaminants, making it a viable option for portable and affordable water

purification compared to untreated water.

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Significance of the Study

The result of this study will benefited the following:

Students. This study will teach students about the purification of water through proving how

unfiltered water can still be purified, as well as the importance of conducting research in gaining

a deeper understanding of how nature-gifts can contribute to societal improvement.

Rural Communities. Many rural areas especially in eastern samar with almost no access to clean

water due to lack of resources. This research offers a sustainable, low-cost solution for water

purification using locally available natural resources (Moringa seeds, Jute leaves, and Guava

leaves).

Local Farmers and Herbalists. This method of water purification promotes the use of locally

available, sustainable materials, encouraging rural farmers to engage in eco-friendly practices for

a more sustainable agriculture.

Conceptual Framework
Figure 1. Conceptual Framework on Utilizing Moringa Seeds (Moringa oleifera), Jute
Leaves (Corchorus olitorius), and Guava Leaves (Psidium guajava) for Portable Water
Purification: LeafFusion Pouch.

Figure One illustrates the independent and dependent variables and their flow in

relation to the study. The first block indicates the types of natural-plant-based coagulants

with subcategories: Moringa oleifera (Malunggay) seeds and Corchorius Olitorius (Jute) leaves.

The second block indicates the natural-plant-based anti-bacteria material with sub category:

Psidium Guajava (Guava) Leaves. The two stated natural-plant-based coagulants and plant-based

anti-bacterial material will be blended and will be utilized as independent variables to determine

their effects as coagulants and purifiers on unfiltered water. The two natural-plant-based

coagulants will significantly impact the dependent variable, the turbidity state of the wastewater,

accordingly manifesting significant results.

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Scope and Limitations

In the context of this study, we acknowledge that this research is not without its

limitations. Even if there is an existing limitations, we came up with the solutions for it to be

resolved and these are how moringa seeds, jute leaves, and guava leaves can be used to filter and

clean dirty water, especially in areas near rivers or places with polluted water, to keep the

citizens safe with good surroundings. We’ll test how well these plant materials work to remove

things like dirt, bacteria that can cause severe diseases on the citizens near the areas.

The study will examine several plant material preparation techniques, including crushing,

drying, and how these techniques might be applied to water purification. Additionally, it also

compares how well these plants remove dirt, mix particles together, and filter water. The study

will also look at how economical and environmentally beneficial these plant materials are in

comparison to treating water with modern filtration using chemicals.

This study is limited to laboratory tests, so it might not fully represent real-life situations

where the water conditions can change a lot (like dirt levels). We’re mainly testing if the plants

can remove dirt and bacteria, further research is needed to determine if they can get rid of other

pollutants like heavy metals or chemicals.

This study is managed on a small scale, so the findings may not apply to large-scale water

treatment. The effectiveness of the plants depends on the preparation method. Lastly, the water

purified by these plants in this study may not be considered safe for drinking, as it might put

everyone’s life in danger as this study is not suitable for reconsidering the health of the citizens

as it might be dangerous for everyone.

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 CHAPTER II

REVIEW OF RELATED LITERATURE

Presented in this chapter is an in-depth review of the literature and studies relevant

to the researchers' investigation of how a natural plant-based blend of Moringa oleifera

seeds, Corchorus olitorius leaves, and Psidium guajava leaves can be used to treat

unfiltered water as a coagulant. This review encompasses scholarly articles and studies

that examine the properties, applications, and potential of these plants, with a particular

emphasis on their role and efficacy in water purification processes.

Studies of Moringa Seeds

It is common these days that there are issues of how many people are exposed to

dangerous levels of chemical and contaminants water pollution in water used for daily life

particularly in rural communities due to lack of modern filters/water filtration. It has been found

that Moringa seeds is the best natural coagulant discovered yet, that can replace aluminium

sulfate (Alum) which is used widely all around the world for purification. This will be of great

help as an alternative and is locally grown in the Philippines and other tropical countries where

one can easily obtain and make use of it.

The application of Moringa Oleifera seed extracts as a natural coagulant has been

investigated recently, and the results indicate great promise to improve the quality of water. The

seeds of Moringa oleifera possess antimicrobial and natural coagulant properties, which make

them potential candidates for water treatment. Water treated with both paste and a 20mL/L

solution of Moringa Oleifera seed extract was clarified to high quality with low turbidity. The

cationic proteins from the seed coat help by binding to negatively charged particles, improving

the seeds' natural ability to remove impurities. Water containing 35mL/L of Moringa seed extract

has proven to be exceptionally capable of significantly effective pollutant removal due to optimal

pH levels within the 6.5-7.5 pH range which are necessary for coagulant activity. Therefore, by

conserving Moringa seeds, which contain nutrients such as phosphate, iron, and fluoride, allows

the seeds to be valuable resources in areas where conventional water treatment is a challenge.

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 The study focuses on “Water purification improvement using moringa Oleifera seed

extract pastes for coagulation follow scoria filtration” this study uses a Volcanic Rocks as filter

medium and including Moringa Oleifera Seeds which is from a natural plant as

coagulant/flocculant. This study is somewhat similar to our own study since we’re also using

Moringa seeds as water purifiers including Jute Leaves and Guava leaves as an anti-bacterial.

Instead of using Volcanic Rocks we're using a combination with 3 different plants (Jute & Guava

leaves and Moringa Seeds) and creating a tea bag. Moreover, the aim of these studies is to

enhance the water purification process to make a cleaner environment.

Studies of Guava Leaf

In this study, guava leaf extract is investigated as a potential wastewater treatment

product, focusing on common water quality issues, such as turbidity and different bacteria. Water

samples containing guava leaf extract have been found to contain reduced turbidity and a

reduction in suspended solids. This effect is likely attributed to guava leaves' high levels of

coagulating agents, including polyphenols, tannins, and flavonoids. Compounds like these

destabilize colloidal particles in water, causing them to aggregate into larger particles that can be

filtered out easily.

This study states that Guava leaf extract is not only known for its coagulating abilities but

also its notable antibacterial properties. This is mainly due to the presence of bioactive

compounds like quercetin, gallic acid, eugenol, and tannins. Research has indicated that these

elements can hinder the proliferation of various bacterial strains. Quercetin and gallic acid

possess antioxidant and antimicrobial properties that affect bacterial cell membrane structures,

diminish bacterial adhesion, and block the expression of virulence factors. On the other hand,

eugenol, which is a phenolic compound, exhibits robust antibacterial effects by compromising

bacterial cell walls and inhibiting the synthesis of proteins.

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 The effectiveness of guava leaf extract in combating bacteria has been evaluated against

several harmful bacterias that can be found in unfiltered bodies of water, such as Coliform

bacteria and E. coli bacteria are the two most common water tests done on wells and springs, to

determine their microbiological safety for drinking. Total coliform bacteria are a large group of

bacteria that are common in surface water, while E. coli bacteria come from the feces of animals

or humans(Bryan Swistock., Jan 30 2019). The extract’s capacity to eradicate or diminish these

dangerous bacteria positions it as a strong candidate for implementation in water purification

systems, particularly in areas with limited access to clean water.

In particular, guava leaves contain a high amount of natural antioxidants, which enhance

their antimicrobial properties and provide extra health benefits when utilized in unfiltered water

of wells and springs. The non-toxic and safe characteristics of guava leaf extract make it a

compelling choice for purifying water, especially in regions where the use of synthetic chemicals

is not preferred because of their possible detrimental impacts on both the environment and

human health.

Studies of Jute Leaves

In the study of Baldoza et al., (2018), it is pointed out that the flocculating ability of

Saluyot (Corchorus olitorius) can be a significant tool to increase the sustainability of treating

turbid river water, particularly the water from Davao River (specifically Bankerohan area) which

is particularly known to be high in pathogenic organisms, turbidity and harmful chemicals due to

the dense population and industrialization around it. Corchorus olitorius (Jute) leaves are

beneficial for water coagulation processes because they have a high content of water-soluble

mucilage, which acts as a natural coagulant and enables them to effectively clump together

suspended particles in water (Lacang et al., 2023; Banerjee et al., 2005). Jute leaf extracts can be

used to treat turbid water by adding them to the water, allowing the mucilage to bind to

suspended particles, which then settle to the bottom as flocs (Yamazaki et al., 2009)

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 Jute leaves have natural coagulation, flocculation and purifies of the pollutants, jute

leaves can help to detoxify the water, clearing the unfiltered water and improves the water

quality. Their leaves are consumed for their nutritive and medicinal values. Several studies

reported that Tossa jute had antiviral, antibacterial and antioxidant activities (Ramadevi, &

Ganapaty, 2011; Barku et al., 2013; Ben Yakoub et al., 2018), due to its high amounts of vitamin

E, β-carotene, ascorbic acid, α-tocopherol, glutathione and phenolic compounds (Furumoto et al.,

2002; Zeghichi et al., 2003; Azuma et al., 1999). The purpose of this study is to show how jute

leaf polysaccharides may be important to health, especially when used to treat infectious bacteria

or in water purification applications where they may lessen bacterial contamination. This

supports our study since Jute leaves has natural coagulants that can help to purify water.

Hypothesis

This study proposes that the LeafFusion Pouch, by combining Moringa seeds, Jute

leaves, and Guava leaves, can effectively purify contaminated water by removing microbial

contaminants, reducing turbidity, and improving overall water quality, making it safe. The

combined plant components will have a synergistic effect that increases the overall water

purification process, out–performing the use of each plant component alone. Specifically, this

study proposes that Moringa seeds, as demonstrated in previous studies (e.g., "Water purification

improvement using Moringa Oleifera seed extract pastes for coagulation followed by scoria

filtration"), will act as an effective natural coagulant, flocculating suspended particles and

microbial agents, thereby significantly reducing turbidity and microbial load in contaminated

water.

Also, Guava leaves, as detailed in the second article ("The Antibacterial and Coagulant

Activity of Psidium Guajava Leaves Extract in Purification of Wastewater"), will contribute both

coagulant and antibacterial properties, further enhancing the removal of pathogens and

improving the microbial safety of the treated water. Lastly, Jute leaves, which have shown

promise in previous studies for their potential coagulant properties, will add to the overall

coagulation process, further supporting the aggregation and removal of impurities in the water.

With these research findings, we may conclude that our study is effective.

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 CHAPTER III

METHODOLOGY

This chapter talks about the methodologies that will be used in the scope of the study. It

exhibits the techniques and identifying procedures involved in collecting data.

Materials

The materials that will be utilized in this research are the following: Moringa oleifera

seeds, Corchorus olitorius leaves, and Psidium guajava folium leaves, mortar and pestle ,

unfiltered water, cloth pouch, thread, beakers (at least three), pH test strips to measure pH levels,

spectrophotometer for turbidity measurement, Microbial load testing (ddPCR for more precise

measurement DNA extraction kit, primers specific to target microorganisms, fluorescent probe,

ddPCR reaction mix, a droplet generator, a ddPCR amplification system, PCR tubes or plates,

pipettes and tips, a thermal cycler, and positive and negative controls.). Vacuum Filtration

[Equipment: Vacuum pump, Vacuum flask, Buchner funnel, Rubber tubing, Filter paper or cloth

(if additional filtration is needed)]. Personal Protective Equipment (PPE).

Set-up

In this experiment, student researchers will utilize the necessary materials and laboratory

equipment in testing the water quality parameters so that accurate results can be achieved. Such

water samples will be collected from locations which have unfiltered water like wells and label

containers for easy tracking. Researchers will follow standard procedures, wear appropriate

protective gear or Personal Protective Equipment(PPE) and ensure safety guidelines are

followed. All data will be recorded carefully, equipment will be cleaned after use or thrown if

necessary, and any remaining samples will be stored properly.

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Flowchart of Experimental Procedure

Figure 2. Procedure Flowchart for treating unfiltered water using plant-based materials

such as Jute Leaves, Guava Leaves, and Moringa Seeds.

Experimental Procedure:

In preparation of materials, collect Moringa Seeds, Jute Leaves, and Guava Leaves. Then,

place the organic material in a clean container and dry under the sun. Next, Crush the Moringa

seeds into a slightly fine powder, and crush the Guava Leaves and Jute Leaves into small pieces.

Lastly, place the prepared materials into an empty cheesecloth pouch and fill it and form it into a

pouch -like shape.

1. Gathering of Plants

The researchers will gather plants for this study, this includes: Moringa Seeds

(Moringa oleifera), Jute Leaves (Corchorus olitorius), and Guava Leaves (Psidium

guajava) through known gardens locally in Tacloban City. For Moringa seeds, the

students will need one hundred and fifty moringa seeds in 1 ounce, which is about one

hundred fifty grams locally. For Jute leaves, the student researchers will need at least

hundred fifty grams. Lastly for Guava leaves, the student researchers will obtain at least

hundred and fifty grams as well.

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2. Acquiring Water Samples

For acquiring water samples, first, the students researchers will collect water

samples from ponds, or wells located in rural areas. Separate water into two different

containers and in equal parts for control and experimental setups.

3. Moringa Seeds, Jute Leaves, and Guava Leaves Preparations

The student researchers will first collect the three types of leaves: moringa seeds,

jute leaves, and guava leaves, then thoroughly wash them to remove any dirt or

impurities, ensuring the leaves are clean. The leaves will then be dried for 1 to 2 days in a

clean area. After the initial drying period, the leaves will be washed again to remove any

remaining residues and then dried for an additional period. Once fully dried, the leaves

will be crushed separately, and the crushed materials will be carefully combined. The

three types of leaves will then be placed into a cloth, which will be securely tied to form

the LeaFusion Pouch, ready for use in water filtration.

4. LeaFusion Pouch Filtration Process

a. Submerge the LeaFusion pouch in the water sample to use it to steep in the water and stir

in a gentle way for uniform mixing and ensure that the filtration pouch has consistent

contact with the water.

b. Allow the pouches to steep in the water for a small to moderate volume of water, from

1-4 hours. During this time, the natural materials in the pouches start to act and filter out

suspended particles and microorganisms.

c. For the vacuum filtration process, set up the necessary equipment by placing the water

sample into a vacuum filtration flask. Position a Buchner funnel on top of the flask and

insert the filtration pouch containing Moringa, Jute, and Guava materials into the funnel.

Connect a vacuum pump to the filtration flask, then activate the vacuum pump to create

suction.This suction will draw the water through the filtration pouch more quickly,

significantly speeding up the filtration process compared to gravity filtration

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d. Observe and record the differences in any sedimentation, water clarity, and odor.

e. Once the filtration period is done, remove the pouch from the container and discard the

materials. Then, filtered water will be ready for quality testing.

5. Testing of Water Quality

a. Measuring the pH by using pH strips, Turbidity can be measured using Microbial Load

using test kits or laboratory assistance of the control and treated water samples.

b. After the filtration process is done, immediately conduct pH measurement as pH can

fluctuate overtime with microbial and chemical changes. The pH of the water samples

will be using commercially available kits like pH strips to measure pH meter (This

method is preferred due to its accuracy and precisions.). The meter will be calibrated

using standard buffer solutions before use to ensure reliable readings.

c. Then, the turbidity will be quantified using a spectrophotometer (measuring at a

wavelength of 680 nm, or as per the spectrophotometer manufacturer’s guidelines for

turbidity). If a spectrophotometer is unavailable, turbidity test kits that use nephelometric

principles can be used.

d. Quantifying microbial load in water samples using droplet digital PCR (ddPCR). DNA

will be extracted from both control and treated water samples, and a reaction mixture

with primers for target microorganisms (e.g., E. coli) and a fluorescent probe will be

prepared. The student will amplify the samples in a droplet generator, which partitions

the reaction into micro-droplets. After amplification, the ddPCR system will count

positive droplets to determine the presence of microbial DNA. The microbial load will be

expressed as copies of DNA per milliliter of water.

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Data Collection and Analysis

After the whole experimentation, the researchers will analyze the results of the

experiment and this will involve detailed comparison of before and after filtration using

the Leafusion Pouch whether the filtration successfully reduced the turbidity. The student

researchers will evaluate the effectiveness of the filtration process by comparing the

recorded data to clean water standards. Noting improvements in water clarity, microbial

depletion, and any additional observations . The findings of this study will help the

researchers better understand whether the Leafusion pouch can be a better and low-cost

solution to provide cleaner water for communities, especially in rural communities.

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Timeline:

1st Week During the first two days, the researchers will begin gathering the
items required for the experiment, which include a food processor,
unfiltered water, a cloth pouch, thread, at least three beakers, and a
spoon and set them aside to be used in the experiment proper.
Within three to four days of preparation, the researchers will gather
moringa seeds, jute leaves, and guava leaves. On days 5 to 7, after
collecting the leaves, the researchers begin to wash with plenty of
water. After rinsing and dipping for 5 minutes, transfer the leaves
to a tray and take it outside for drying. The leaves will be dried for
1-2 days. Repeat this method until the color and flavor of the
leaves are discarded.

2nd Week On day eight, the drying proceeds. The researchers will afterwards
prepare the pouch's materials, which include cloth and a rubber
band. On days 9 to 10, the dried leaves were washed on a tray with
constant pressure to remove its natural color and dry it for 5 more
hours. On day 11-14, the researchers will collect the dried leaves
and begin crushing them with a food processor. After crushing the
leaves, the researchers will place them in a cloth and bind it with a
rubber band to form the Leafusion Pouch.

3rd Week On days 15-16, the researchers will prepare for the experiment by
gathering the necessary items such as beakers, unfiltered water,
cloth, and a rubber band for cover. On days 17-21, the researchers
will conduct the experiment by placing the Leafusion Pouch in
water and wait for 5 days. During these days, the researchers will
monitor the progress of the experiment and assure its safety.

During the final week of conducting this research, the researchers

will focus thoroughly examining and documenting the changes of
the filtration process to assess the Leafusion pouch effectiveness to
4th Week lessen the turbidity of the water. This will involve detailed
comparison of before and after filtration using the Leafusion Pouch
whether the filtration successfully reduced the turbidity. The
findings of this study will help us better understand whether the
Leafusion pouch can be a better and low-cost solution to provide
cleaner water for communities especially in rural communities. In
this week, the researchers will be using pH strips for the pH level
of the water, a spectrophotometer for the water's turbidity and if a
spectrophotometer is not available, the researchers will be asking
for laboratory assistance. For microbial load, we will be asking for
laboratory assistance.

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Bloetscher, F., Meeroff, R., Roblyer, A., Prymes, J., Daniel, A., Julia, B., & Angela, C. (n.d.).
Algal control in warm weather ponds using EMOH device. Retrieved from
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Jung, A. -V., Le Cann, P., Roig, B., Thomas, O., Baurès, E., & Thomas, M. -F. (2014).
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Rocker Scientific. (2024). What is vacuum filtration? | Select the right vacuum filtration
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