Muntingia Calabura

1
Characterization of Powdered Muntingia calabura L. (Sarisa) Leaves as Potential
Biosorbent for the Removal of Copper sulfate and Zinc sulfate from Aqueous
Solutions
ASHLIE JANE B. BALIDIO
MARIA ISABEL T. GARCIA
JB SHIELA I. MUNIEZA
Student Researchers
PHILLIP RAYMUND DE OCA
Project Adviser
BACOLOD CITY NATIONAL HIGH SCHOOL
Bacolod City, Negros Occidental
Region VI, Western Visayas

2
Abstract
Heavy metal pollution is an ecological threat that affects water quality for viable
use. Freshwater sources significantly decreased over the years while the demand for
water-intensive goods continually rise. Heavy metals are characterized as natural
components of the earth that induce toxicity even at low concentrations. Consequently,
there is a need to come up with novel methods of heavy metals remediation which are
more effective and reliable. Therefore, the present study sought to determine the rate of
biosorption of copper sulfate and zinc sulfate from wastewater using powdered
Muntingia calabura Linn (Sarisa) leaves in aqueous solutions prepared in different
biosorbent doses and contact times in terms of: pH level, turbidity, and Total Dissolved
Solids. The leaves of the plant were oven dried and grinded to obtain its powdered state.
Varied amount of biosorbent doses such as 0.2, 0.5 and 0.8 grams were subjected with
different contact times of 5, 15 and 25 minutes for each chemical solution.Between
varied biosorbent doses, insignificant differences were obtained in the results based on
themajority of set-ups prepared.On the other hand, significant relationships were obtained
between pH level and TDS. It is also the same with pH and turbidity having an inverse
proportion. The removal capacity increases as the biosorbent dose rises. Hence, the initial
and shortest exposure time showed optimum results and the rest showed declining rates
as time was added. In conclusion, Muntingiacalabura is a low-cost biosorbentexhibiting
low biosorptive potential against copper and zinc sulfate from aqueous solutions.
3
Table of Contents
Title Page 1
Official Abstract Form 2
Table of Contents 3
List of Figures 4
List of Tables 8
Research Plan 10
Introduction 16
Methodology 21
Results and Discussion 27
Summary of Findings 69
Conclusion 72
Recommendation 73
Acknowledgment 74
References 75
Appendices Cost Analysis 77
Documentations 78
Curriculum Vitae 81
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List of Figures
Figure 1. Effect of Varying Biosorbent Dose with 5 minutes Contact on pH Level for
Copper sulfate
Figure 2. Effect of Varying Biosorbent Dose with 5 minutes Contact on Turbidity
for Copper sulfate
Figure 3. Effect of Varying Biosorbent Dose with 5 minutes Contact on TDS for
Copper sulfate
Figure 4. Effect of Varying Biosorbent Dose with 15 minutes Contact on pH Level
for Copper sulfate
for Copper sulfate
Copper sulfate
for Copper sulfate
for Copper sulfate
Copper sulfate
for Zinc sulfate

5
for Zinc sulfate
Zinc sulfate
for Zinc sulfate
for Zinc sulfate
Zinc sulfate
for Zinc sulfate
for Zinc sulfate
Zinc sulfate
Figure 19. PH Level Mean Comparisons with Varying Biosorbent Dose and Contact
Time for Copper sulfate
Figure 20. Turbidity Level Mean Comparisons with Varying Biosorbent Dose and
Contact Time for Copper sulfate

6
Figure 21.TDS Level Mean Comparisons with Varying Biosorbent Dose and
Time for Zinc sulfate
Contact Time for Zinc sulfate
Figure 24.TDS Level Mean Comparisons with Varying Biosorbent Dose and
Figure 25. Gathering and weighing of Muntingia calabura leaves
Figure 26. Washing of Muntingia calabura leaves
Figure 27. Weighing of washed Muntingia calabura Leaves
Figure 28. Drying of Muntingia calabura leaves
Figure 29. Weighing of dried Muntingia calabura leaves
Figure 30. Pulverizing Muntingia calabura leaves
Figure 31. Muntingia calabura leaves in powdered form
Figure 32. Oven-drying of powdered Muntingia calabura leaves
Figure 33. Weighing of oven-dried powdered Muntingia calabura leaves
Figure 34. Weighing of 0.2 g of powdered Muntingia calaburaleaves
Figure 35. Weighing of 0.5g of powdered Muntingia calabura leaves

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Figure 36. Weighing of 0.8g of powdered Muntingia calabura leaves
Figure 37. Weighing of 30g Copper sulfate
Figure 38. Weighing of 30g Zinc sulfate
Figure 39. Distilled water
Figure 40. Mixing of 300mL Copper sulfate solution
Figure 41. Mixing of 300mL Zinc sulfate solution
Figure 42. Gathering of Materials and Equipment
Figure 43. Preparation of samples
Figure 44. Centrifugation of prepared samples
Figure 45. Placing sample solutions on the test tube racks
Figure 46. Filtering Samples
Figure 47. Transferring samples to bottle containers
Figure 48. Analyzing water of samples and recording results

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List of Tables
Table 1. Amount of Muntingia calabura powder or biosorbent dose and Contact time
on Copper sulfate in Treatments A, B, and C
on Zinc sulfate in Treatments A, B, and C
Table 3.Analysis of Variance for Copper sulfate with 5 minutes contact time
Table 4. Correlations Between pH, Turbidity and TDS of Copper sulfate with 5
minutes Contact Time
Table 5. Analysis of Variance for Copper sulfate with 15 minutes contact time
Table 6. Correlations Between pH, Turbidity and TDS of Copper sulfate with
15minutes Contact Time
Table 8. Correlations Between pH, Turbidity and TDS of Copper sulfate with
Table 9. Analysis of Variance for Zinc sulfate with 5 minutes contact time
Table 10. Correlations Between pH, Turbidity and TDS of Zinc sulfate with 5
Table 12. Correlations Between pH, Turbidity and TDS of Zinc sulfate with
9
Table 14. Table 7. Multiple Comparisons for Turbidity

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Characterization of Powdered Muntingia calabura Linn (Sarisa) Leaves as Potential
Biosorbent for the Removal of Copper sulfate and Zinc sulfate from Aqueous
Solutions
Proponents: Ashlie Jane B. Balidio, Maria Isabel T. Garcia, JB Shiela I. Munieza
Gr. 10-Newton
A. Research Plan/Question or Problem being addressed
Heavy metals are dense metals that can be very toxic even at low concentrations.
They cannot be degraded easily and poses a threat for bioaccumulation; they are natural
occurring elements but have drastically increased when urbanization and industrialization
started (Lenntech n.d.). Two of the most common heavy metals are Copper (II) and Zinc
(II) which are used in wide scale applications for industrial and pharmaceutical purposes
(Thankur, 2013). However, excess copper like copper sulfate leads to nausea and
gastrointestinal problems while zinc in the form of zinc sulfate is a gastrointestinal and
respiratory irritant and overexposure is likely to cause death.
Adsorption for removing heavy metal from wastewater has become one of the
technically important and effective mechanisms used. Nevertheless, its application fields
are restricted due to its expensiveness and limited scope (Arvanitoyannis and Varzakas
2008). For this reason, the researchers will apply the use of agricultural wastes such as
Muntingia calabura Linn (Sarisa) leaves as a novel sorbent in the removal of copper
sulfate and zinc sulfate from aqueous solution.

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B. Goals / Expected Outcomes/ Hypothesis
Generally, the main objective of this study is to determine the Copper Sulfate and
Zinc Sulfate Biosorbent potential of powdered Muntingia calaburaLinn (Sarisa) leaves in
Aqueous Solutions.
Specifically, it seeks to answer the following objectives:
1. What is the Copper sulfate and Zinc sulfate biosorbent potential of powdered
Muntingia calaburaLinn (Sarisa) leaves in aqueous solutions prepared in different
biosorbent dose and contact time and tested in terms of pH level, turbidity, and
TDS (total dissolved solids)?
2. Is there a significant difference in the Copper sulfate and Zinc sulfate biosorbent
potential of powdered Muntingia calabura Linn (Sarisa)leaves in aqueous
solutions prepared in different biosorbent dose and contact time and tested in
terms of pH level, turbidity, and TDS (total dissolved solids)?
3. Is there a significant relationship between the varying biosorbent dose and contact
time of Powdered Muntingia calabura Linn (Sarisa) leaves and its biosorbing
potential of Copper sulfate and Zinc sulfate in terms of pH level, turbidity and
TDS?
C. Procedures
C.1 Gathering of Materials
● The researchers will gather the materials needed in making the powdered
Muntingia calabura Linn (Sarisa) leaves: 600g of Muntingia calabura (Sarisa)
leaves, grinder or blender, oven, mortar and pestle, airtight container.

12
● Muntingia calabura Linn (Sarisa) Leaves will be collected from the residence of
Mr. Ricardo H. Gonzales at corner Handumanan access Rd. and Jupiter Streets,
Hillside Subdivision, Brgy. Mansilingan, Bacolod City. It will be verified at
Bureau of Plant Industry and Quarantine Services, Bacolod City.
● The materials needed to make the Copper sulfate solution (100,000 ppm) and
Zinc sulfate solution (100,000 ppm): 1 gram of copper sulfate will be dissolved in
10 ml distilled water and 1 gram of zinc sulfate will be dissolved in 10 ml distilled
water and will be used for all experiments with required dilution.
● The researchers will use 600mL of distilled water for this set-up.
● During the experiments the researchers will gather the things needed such as:
eighteen (18) 0.2g, 0.5g and 0.8g of Muntingia calabura Linn (Sarisa) Leaves,
fifty-four (54) 10 mL test tubes, two (2) 500 ml beaker, two (2) graduated
cylinders, centrifuge, two (2) stirring rods, filter papers, funnels, test tube racks,
an oven and a digital weighing scale.
C.2 Preparation and Carbonization of Muntingia calabura Linn (Sarisa) Leaves
● In this study, Muntingia calabura Linn (Sarisa) leaves will be used as the sorbent
for the removal of copper and zinc. The mature Muntingia calaburaLinn leaves
will be collected. They must be washed thoroughly with clean water and be rinsed
with distilled water to remove impurities.
● The leaves will then be air-dried for 20-30 days
● The dried leaves will be crushed and be formed to powder by using a blender or
grinder. It will again be pounded using mortar and pestle. It will be sieved and
13
oven-dried to remove additional moisture. It will then be stored in an airtight
container for further use to avoid contact with moisture from the atmosphere.
C.3 Preparation of Copper solution and Zinc Solution (100,000 ppm)
● Sample Preparation of Copper solution (100,000 ppm) will use 30 grams of
copper sulfate in 300 ml distilled water
● Sample Preparation of Zinc solution (100,000 ppm) will use 30 grams of zinc
sulfate dissolved in 300 ml distilled water.
C.4 Testing the Biosorptive Potential of Powdered Muntingia calabura Leaves
C.4.1 Effect of Biosorbent dose
● To find out the effect of biosorbent dose for the biosorption of Copper Sulfate and
Zinc Sulfate from aqueous solution, biosorption studies will be carried out by
varying the amount of biosorbents (0.2, 0.5, 0.8g) while using initial metal
concentration (100,000 ppm) of 10ml Copper Sulfate and Zinc Sulfate and
centrifuge with varied contact times replicating three (3) times for each unit.
C.4.2 Effect of Contact time.
● The effect of contact time on biosorption will be studied between 10 min., 15
min., and 20 min. of centrifugation. The experiment will be carried out using
10ml of Copper Sulfate solution (100,000 ppm) and 10ml of Zinc Sulfate
(100,000ppm) solution containing varied biosorbent doses for each treatment,
replicating three (3) times for each unit.

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C.5 Analysis
● After centrifugation, the samples will be filtered on another flask or beaker and
will be transferred to clean bottles to be analyzed in BACIWA Laboratory.
● The samples obtained from the different treatments will be brought to BACIWA
Laboratory to analyze pH level, turbidity and Total Dissolved Solids using pH
meter for pH level, turbidimeter for turbidity and cond meter for TDS.
● For convenient sampling, each sample of 10 mL treated solution will be mixed
with distilled water to reach the amount of 500 mL water sample needed to be
tested in BACIWA. Therefore, will be submitting with a 2000 ppm chemical
concentration.
C.6 Statistical Tool
● Mean and Standard Deviation will be used to determine the Copper sulfate and
Zinc sulfate biosorbent potential of powdered Muntingia calabura Linn (Sarisa)
Leaves in aqueous solutions prepared in different biosorbent dose and contact
time and tested in terms of pH level, turbidity, and TDS (total dissolved solids).
● ANOVA will be used to determine if there is a significant difference in the
Copper sulfate and Zinc sulfate biosorbent potential of powdered Muntingia
calabura Linn (Sarisa) Leaves in aqueous solutions prepared in different
biosorbent dose and contact time and tested in terms of pH level, turbidity, and
TDS.
● Pearson's r will be used whether there is a significant relationship between the
varying biosorbent dose and contact time of Muntingia calabura Linn (Sarisa)
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Leaves and its biosorptive potential of Copper sulfate and Zinc sulfate in terms of
pH level, turbidity and TDS?
D. References
Arvanitoyannis, Ioannis S., and Theodoros H. Varzakas. Activated Carbon. 2008.
https://www.sciencedirect.com/topics/agricultural-and-biological-
sciences/activated-carbon (accessed September 12, 2018).
Jena, Swarnabala, and Rajesh Kumar Sahoo. “Removal of Pb(Ii) From Aqueous Solution
Using Fruits. 2017.
https://pdfs.semanticscholar.org/f7c3/06d9bb510725c7cabaa0819ca6f2556fa7cb.
pdf (accessed September 2018).
Lenntech. n.d. https://www.lenntech.com/processes/heavy/heavy-metals/heavy-
metals.htm (accessed 2019).
Suhadiyah, Sri, Djamal Sanusi, Samuel Paembonan, and Roland A. Barkey.ijstr.org.
March 2013. http://www.ijstr.org/final-print/mar2013/Lead-Accummulation-
Potential-By-Leaves-With-Abundant--Trichomes-Muntingia-Calabura-L.-And-
Rare-Trichomes-Mimusops-Elengi-L.-In-Makassar-Indonesia.pdf (accessed
September 1, 2018).
Thankur, Lokendra Singh. researchgate.net. 2013.
https://www.researchgate.net/publication/271281628_Heavy_metal_Cu_Ni_and_
Zn_Toxicity_health_hazards_and_their_removal_techniques_by_low_cost_adsor
bents_A_short_overview.
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Introduction
Heavy metal pollution is an ecological threat that affects water quality for viable
use. Heavy metals are relatively high density metals that induce toxicity even at low
concentrations (Lenntech,n.d.). It is one of major factors in the rising ecological threat on
water pollution and has been a subject for global public concerns. These are naturally
occurring elements in Earth’s crust but have drastically increased due to the discharged
from anthropogenic sources such as residential, industrial, metallurgical and agricultural
activities[ CITATION Bra07 \l 13321 ]. Since they are elements, they cannot be neither
degraded nor destroyed easily and possess bio-accumulative properties that affect the
environment and living organisms as well. According to the Environmental Management
Bureau out of 127 freshwater sources in the Philippines, 47% retain good quality and
58% was reported to be of coliform contamination. The State of water Environmental
Issues: Philippines states that the Domestic sector contributes 48% of pollution followed
by agricultural (37%) and industrial (15%) sectors respectively. [ CITATION Sta19 \l
13321 ]
Two most common heavy metals are Copper and Zinc which are used in wide-
scale applications for industrial and pharmaceutical purposes. Copper is a metallic
element that is ductile, malleable and commonly used for electrical applications due to its
high conductivity.[ CITATION Ack77 \l 13321 ] Specifically, copper sulfate is a bluish-white
crystalline powder highly soluble compound used as an algaecide, pesticide and

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molluscicide. It causes eye and skin irritation upon exposure. Inhalation and ingestion of
copper sulfate causes gastrointestinal problems and severe organ damages which later on
may lead to death.[ CITATION Nat191 \l 13321 ] Zinc is also of great industrial importance
and has been extensively mined for galvanizing purposes, protective coating and
component for alloys[ CITATION Ack77 \l 13321 ] . Particularly, zinc sulfate is a white
crystalline powder that is highly soluble in water, as a supplement it helps maintain zinc
levels in human body and herbicide since it inflicts damage in plant cells. Zinc sulfate is a
gastrointestinal and respiratory irritant; it causes alterations in blood chemistry and linked
with depression.[ CITATION Nat19 \l 13321 ] These heavy metals enter water through; waste
streams from metal-manufacturing facilities, corrode pipes, mining discharges, which is
absorbed by the soil, and causes groundwater contamination[ CITATION Ack77 \l
13321 ].These are industrial wastes proven to be toxic especially in aquatic life forms. The
presence of heavy metal ions in water induces bitter-metallic taste, acidic pH level, low
total dissolved solids and high turbidity which adversely affect the ecosystem.
The researchers were mainly influenced by the increasing accumulation levels of
heavy metals in water particularly in freshwater which is necessary for plants, animals
and human survival. Moreover, the rising trend in the global population increases water-
intensive goods demand. Countries classified as under-developed and developing,
experience inadequate supply of safe drinking water for use and for vegetation. Various
approaches have been studied and applied in resolving the issue. Major drawbacks from
conventional treatments were production of toxic sludge, restriction of application fields
due to limited scope and expensiveness of material. Adsorption for removing heavy metal
18
from wastewater has become one of the technically important and effective mechanisms
used. In recent years, demand for economic and inexpensive generation of sorbents from
agro-based products has been widely studied for efficiency [CITATION Placeholder1 \l
13321 ].A process called biosorption is a subcategory of adsorption, characterized by fast
binding of ions in an aqueous solution unto the biomass surface with functional groups
present. It is an efficient, economical and eco-friendly approach on removal of harmful
contaminants based on state of the Art Biosorption Process.
Muntingia calaburaLinn or Sarisa in Visayan dialect is a fast growing tree native in
neo-tropical regions. It is considered a pest in Western countries due to its ability to adapt
on many conditions. It has white flowers, red berry fruit, which is many seeded, and
abundant glandular trichomes in the leaves. Leaf trichomes are hair outgrowth from the
epidermis of a plant. They reflect radiation, lower plant temperature, reduce water loss
and protect plant from pollution including heavy metal toxicity. Nullification and
detoxification of heavy metal on plants happens in the trichomes. Various studies cited
that Muntingiacalabura L. showed remarkable results in uptake of Lead (Pb) in air
[ CITATION Suh13 \l 13321 ] , Chromium (Cr) [ CITATION Aat14 \l 13321 ] and cationic dyes
(methylene blue, methylene red and malachite green) from aqueous solutions.
Consequently, there is a need to come up with novel methods of heavy metals
remediation which are more effective and reliable. For this reason, the researchers studied
and determined the biosorption potential of Muntingiacalabura Linn or Sarisa leaves as a
novel biosorbent in the removal of copper sulfate and zinc sulfate from aqueous solution.
To attest its potential the researchers set pH Level, Turbidity and Total Dissolved
Solids as parameters for the study. The pH level is measured to determine the activity of
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hydrogen ions in an aqueous solution, acidity and solution reaction extant is measured by
pH level. Organic materials are less soluble and more ready for adsorption and pH level
is a good indication that it is getting less of organics because as pH increases the capacity
for removal is getting slower. (Activated Carbon Filtration, 2000) The standard pH levels
of water are those from the range of 6.5 to 8.5. Turbidity is a measure of the degree to
which the water loses its transparency due to the presence of suspended particulates. The
more total suspended solids in the water, the murkier it seems and the higher the
turbidity. Turbidity is measured in NTU or Nephelometric Turbidity Units, The
instrument used for measuring it is called nephelometer or turbidimeter, which measures
the intensity of light scattered at 90 degrees as a beam of light passes through a water
sample. Total dissolved solids (TDS) is a measure of the combined inorganic and organic
substances dissolved in water, which is directly related to the electrical conductivity of
water. It can also be used as an indication of change. Maximum contamination level
(MCL) of 500mg/L (500 ppm) is advised by the EPA Secondary Regulations advise a for
TDS. It is considered unfit for human consumption when it exceeds 1000mg/L. TDS
levels in water can make the water taste salty or bitter and can also cause corrosion of
metal equipment. It also can contribute to skin and eye irritations.
Generally, the main objective of this study is to determine the Copper Sulfate and
Zinc Sulfate Biosorbent potential of powdered Muntingiacalabura Linn (Sarisa) leaves in
Aqueous Solutions.
Specifically, it seeks to answer the following objectives:

20
1. What is the Copper sulfate and Zinc sulfate biosorbent potential of powdered
Muntingiacalabura Linn (Sarisa) leaves in aqueous solutions prepared in different
biosorbent dose and contact time and tested in terms of pH level, turbidity, and TDS
(total dissolved solids)?
2. Is there a significant difference in the Copper sulfate and Zinc sulfate biosorbent
potential of powdered Muntingiacalabura Linn (Sarisa) leaves in aqueous solutions
prepared in different biosorbent dose and contact time and tested in terms of pH level,
turbidity, and TDS (total dissolved solids)?
3. Is there a significant relationship between the varying biosorbent dose and contact
time of Powdered Muntingiacalabura Linn (Sarisa) leaves and its biosorbing potential of
Copper sulfate and Zinc sulfate in terms of pH level, turbidity and TDS?
From the objective raised, the following hypotheses are the Null hypothesis of the
study:
1. There is no significant difference in the Copper sulfate and Zinc sulfate
biosorbent potential of powdered Muntingiacalabura Linn (Sarisa) leaves in aqueous
solutions prepared in different biosorbent dose and contact time and tested in terms of pH
level, turbidity, and TDS (total dissolved solids).
2. There is no significant relationship between the varying biosorbent dose and
contact time of Powdered Muntingiacalabura Linn (Sarisa) leaves and its biosorbing
potential of Copper sulfate and Zinc sulfate in terms of pH level, turbidity and TDS.
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Methodology
A. Research Design
The experiment was laid with eighteen (18) experimental groups (varying dose
and contact time of sorbents): replicated three (3) times; for a total of seventy-four (54)
experimental units. The pH level, turbidity and TDS (Total Dissolved Solids) of treated
Copper sulfate and Zinc sulfate solutions with varying dose and contact time of
Muntingia calabura Linn powder were analyzed and recorded after the application of
intervention. Then, comparisons were made and group differences were used to
determine the effects of treatment.
B. Materials
The researchers manually collected 600g of Muntingia calaburaleaves. Before
experimentation, 30g of Copper Sulfated and 30g of Zinc Sulfate was prepared for 600ml
distilled water, a digital weighing scale andeighteen (18) 0.2g, 0.5g and 0.8g was
prepared. Equipment and materials for making of Muntingia calabura Powder such as:
grinder or blender, mortar and pestle, oven, airtight container; and for making Copper
Sulfate and Zinc Sulfate stock solutions:two (2) 400 ml beaker, stirring rod and digital
weighing scale were prepared. For the experimentation, the researchers gathered the
following: forty-five (45) 10 mL test tubes, (54) solution containers, two (2) graduated
22
cylinders, centrifuge, two (2) stirring rod, filter paper, test tube rack, oven and digital
weighing scale for the experimentation. During water analysis, they used the following
equipment: pH meter, turbidimeter and an electrical conductivity meter (with the function
to also determine the TDS).
C. Location and Duration of the Study
The Muntingia calabura Linn (Sarisa) leaves were collected from the residence of
Mr. Ricardo H. Gonzales at corner Handumanan access Rd. and Jupiter Streets, Hillside
Subdivision, Brgy. Mansilingan, Bacolod City. The leaves were verified at Bureau of
Plant Industry and Quarantine Services, Bacolod City. The stock solution of Copper
Sulfate and Zinc Sulfate were made and were taken from the DOST Building-Stock
room, of Bacolod City National High School, P. Hernaez Extension St., Brgy. Taculing,
Bacolod City, Negros Occidental.
At Colegio San Agustin – Bacolod under the supervision of a laboratory technician,
was where: the powdered Sarisa (Muntingia calabura L.) was oven-dried and weighed;
the Copper Sulfate and Zinc Sulfate was weighed; and it was where the experimentation
was continued by the researchers.
After conducting the experiment, the precipitates were measured and the researchers
went to Bacolod City Water District to physically and chemically characterize the
properties of water treated for its pH, TDS, turbidity and the amount of chemicals present
in the solution (ppm).

23
D. Flowchart of Procedures
Collection of Muntingiacalabura Leaves
Preparation of Muntingiacalabura Leaves
Preparation of Treatments by varying Dosages
Treatment A Treatment B Treatment C

0.2 grams of 0.5 grams of 0.8 grams of
Powdered Powdered Powdered
MuntingiacalaburaLea Muntingiacalabura MuntingiacalaburaLe
ves Leaves aves
Preparation of Aqueous Solutions
Adding Copper sulfate Adding Zinc sulfate into

into Distilled Water Distilled Water
Mixing of the Solutions
Experimentation
Testing Capacity of Treatments for Varying Contact Time
Set-up 1 (5 min) Set-up 2 (15 min) Set-up 3 (25 min)

Trial 1 Trial 1 Trial 1
24
Analysis of Water
pH Level Total Dissolved Solids Turbidity
E. General Procedure
Gathering of Results and Statistical Analysis
E.1 Gathering of Materials
The researchers gathered the materials needed to make the powdered Muntingia
calaburaleaves, eighteen (18) 0.2g, 0.5g, 0.6g were needed. Stock solutionsof 30g
Copper Sulfate and 30g Zinc Sulfate with (100,000 ppm) or 100,000 mg/L concentration
was mixed to 300mL distilled water for each solution. Then, the materials and equipment
needed were gathered for experimentation.
E.2 Preparation and Carbonization of PowderedMuntingia calabura (Sarisa) Leaves
In this study, Muntingia calabura Linn (Sarisa) leaves was used as the bioorbent for
the removal of copper and zinc. The mature Muntingia calabura leaves were collected.
They were washed thoroughly with clean water and rinsed with distilled water to remove
impurities. The leaves were then be air-dried for 20-30 days
The dried leaves were crushed and formed to powder by using a blender and grinder.
It was again be pounded using mortar and pestle. It was sieved and oven-dried to remove
additional moisture. It was then stored in an airtight container for further use to avoid
contact with moisture from the atmosphere.
E.3 Preparation of Copper solution and Zinc Solution (100,000 ppm)

25
The sample preparation of Copper solution (100,000 ppm) used 30 grams of
Copper sulfate in 300 ml distilled water.
Then the sample preparation of Zinc solution (100,000 ppm) used 30 grams of
Zinc sulfate dissolved in 300 ml distilled water.
E.4 Testing the Biosorptive Potential of Powdered Muntingia calabura (Sarisa)
Leaves
E.4.1 Effect of Biosorbent dose
To find out the effect of biosorbent dose for the biosorption of Copper Sulfate and
Zinc Sulfate from aqueous solution, biosorption studies were carried out by varying the
amount of biosorbents (0.2, 0.5, 0.8g) while mixing it to a metal concentration (100,000
ppm) of 10ml Copper Sulfate and Zinc Sulfateand centrifuge while using its’ assigned
contact time between 10 min, 15 min, 20 (Jena and Sahoo 2017).
E.4.2 Effect of Contact time.
The effect of contact time on biosorption was studied between 10 min., 15 min.,
and 20 min. of centrifugation. The experiment was carried out using 10ml of Copper
Sulfate solution (100,000 ppm) and 10ml of Zinc Sulfate (100,000ppm) solution by
mixing the assigned biosorbent dose between 0.2, 0.5 and 0.8 on the chemical solutions
(Jena and Sahoo 2017).
E.5 Water Analysis
After centrifugation, the samples were filtered on another flask or beaker and were
transferred to clean bottles to them in BACIWA.The samples obtained from the different
26
treatments were brought to BACIWA Laboratory to test its pH level, turbidity and Total
Dissolved Solids using different laboratory apparatuses such as pH meter for pH level,
turbidimeter for turbidity and conductivity meter for TDS(Total Dissolved Solids).For
convenient sampling, each sample of 10 mL treated solution were mixed with distilled
water to reach the amount of 500 mL water sample needed to be tested in BACIWA.
Therefore, submitted with a 2000 ppm chemical concentration.
E.6 Analysis of Statistical Data
Mean and Standard Deviation were used to determine the Copper sulfate and Zinc
sulfate biosorbent potential of powdered Muntingia calabura Linn (Sarisa) leaves in
aqueous solutions prepared in different biosorbent dose and contact time and tested in
terms of pH level, turbidity, and TDS (total dissolved solids).
ANOVA was used to determine if there is a significant difference in the Copper
sulfate and Zinc sulfate biosorbent potential of powdered Muntingia calabura Linn
(Sarisa) leaves in aqueous solutions prepared in different biosorbent dose and contact
time and tested in terms of pH level, turbidity, and TDS (total dissolved solids).
Pearson's r was used whether there is a significant relationship between the varying
biosorbent dose and contact time of Powdered Muntingia calabura Linn (Sarisa)leaves
and its biosorbing potential of Copper sulfate and Zinc sulfate in terms of pH level,
turbidity and TDS.

27
Results and Discussion
This section presents the data gathering procedure in connection with the purpose
of the experimentation and for the investigation of the researchers, and the descriptive
presentation of data based on its turbidity, pH level and total dissolved solids.
Table 1 below shows the amount of biosorbent dose, water (ml) used, the amount
of copper sulfate (g) and contact time in treatments A, B and C.
on Copper sulfate in Treatments A, B, and C
Amount of Amount of Amount of Contact
Biosorbent Dose Water Copper sulfate Time
(g) (mL) (g) (min)

Treatment A.1 0.2 10 1 5
Treatment B.1 0.5 10 1 15
Treatment C.1 0.8 10 1 25
28
Table 2 below shows the amount of biosorbent dose, water (ml) used, the amount
of zinc sulfate (g) and contact time in treatments A, B and C.
on Zinc sulfate in Treatments A, B, and C
Amount of Amount of Amount of Contact
Biosorbent Dose Water Zinc sulfate Time
(g) (mL) (g) (min)

Figure 1 below, shows the effect of varying biosorbent dose with 5 minutes contact
time on pH Level per trial. The neutral pH of water is 7, value lesser than 7 is considered
acidic and value greater than 7 is considered basic. The determination of pH Level use
Electrometric method through pH meter device. Samples with 0.2g of powdered
Muntingia calabura exhibited a mean of 4.23, 0.5g of powdered Muntingia calabura has
29
a mean of 4.45 and 0.8g has a mean of 4.47 and as the grams increases the pH level also
increases.
Figure 1. Effect of Varying Biosorbent Dose with 5 minutes Contact on pH Level for
Copper sulfate
Copper sulfate 5mins

4.80
4.70
4.60
4.50
pH Level
4.40
4.30
4.20
4.10
4.00
3.90
0.2g 0.5g 0.8g
30
time on Turbidity per trial. The results were determined with the use of turbidimeter. The
sample with 0.2g biosorbent dose had a mean result of 0.64 as the highest while the
second sample with 0.5g obtained 0.36 mean which was the lowest and the trials
containing 0.8g garnered 0.55.

31
for Copper sulfate

1.20
1.00
0.80
Turbidity
0.60
0.40
0.20
0.00
0.2g 0.5g 0.8g
32
Figure 3 below, shows the effect of varying biosorbent dose with 5 minutes
contact time on TDS (Total Dissolved Solids). The results were measured through the use
of Electrical Conductivity (EC) meter. There was a decreased in amount of TDS from the
sample containing 0.2g of biosorbent dose which had a mean of 361.33, 0.5g sample
obtained 281.93 mean and 0.8g had the lowest result of 255.27.
Copper sulfate

450.00
400.00
Total Dissolved Solids
350.00
300.00
250.00
200.00
150.00
100.00
50.00
0.00
0.2g 0.5g 0.8g
33
Table 3 shows the Analysis of Variance for copper sulfate with Five (5) minutes
contact time. The significance level is set to p ≤ 0.05, value lower or exact to 0.05 implies
rejection of null hypothesis and value higher than 0.05 implies that results failed to reject
null hypothesis. The pH level has a significance of 0.169 which is non-significant.
Turbidity has a significance of 0.605 which is also nonsignificant. Total Dissolved Solids
has a significance value of 0.684 which is also non-significant.
Table3. Analysis of Variance for Copper sulfate with 5 minutes contact time
ANOVA
Sum of Squares Mean Square F Sig.
Between Groups .102 .051 2.431 .169
pH Within Groups .126 .021
Total .228
Between Groups .124 2.882 .546 .605
Turbidity Within Groups .684 2.149
Total .808
Between Groups 18265.609 9132.804 .405 .684
TDS Within Groups 135397.520 22566.253
Total 153663.129
Table 4 shows the correlations for copper sulfate with 5 minutes contact time. The
significance level is set to p ≤ 0.01, value lower or exact to 0.01 implies rejection of null
hypothesis and value higher than 0.01 implies that results failed to reject null hypothesis.
34
Only between pH level and TDS obtained a p-value of 0.001 which is lower than the
alpha value of 0.01, implicating that it has a significant relationship. The rest showed
insignificant values such as: 0.083 between TDS and turbidity; and 0.080 between pH
and turbidity.
Correlations
pH Turbidity TDS
pH Pearson Correlation 1 -.612 -.899**
Sig. (2-tailed) .080 .001
N 9 9 9
Turbidity Pearson Correlation -.612 1 .607
Sig. (2-tailed) .080 .083
N 9 9 9
**
TDS Pearson Correlation -.899 .607 1
Sig. (2-tailed) .001 .083
N 9 9 9
**. Correlation is significant at the 0.01 level (2-tailed).
contact time on pH Level per trial. The neutral pH of water is 7, value lesser than 7 is
considered acidic and value greater than 7 is considered basic. The determination of pH
Level used Electrometric method through pH meter device. Samples with 0.2g of
powdered Muntingia calabura exhibited a mean of 4.19 as the lowest, 0.5g of powdered
Muntingia calabura has a mean of 4.29 and 0.8g has a mean of 4.30as the highest and as
the biosorbent dose increases the pH level also increases.

35
for Copper sulfate

4.35
4.30
4.25
4.20
pH Level
4.15
4.10
4.05
4.00
0.2g 0.5g 0.8g
on Turbidity for Copper Sulfate per trial. The results were determined with the use of
turbidimeter. The sample with 0.2g biosorbent dose had a mean result of 0.50, the second
sample with 0.5g obtained 0.33 mean which was the lowest and the samples containing
0.8g garnered 1.00 which is the lowest.
for Copper sulfate

36

2.00
1.80
1.60
1.40
1.20
Turbidity
1.00
0.80
0.60
0.40
0.20
0.00
0.2g 0.5g 0.8g
contact time on Total Dissolved Solids (TDS). The results were measured through the use
of Electrical Conductivity (EC) meter. There was a decreased amount of TDS from the
sample containing 0.2g biosorbent dose which had a mean of 380.67, 0.5g sample
obtained 342.33 mean and increased of 347.33 mean on the 0.8g sample.
Copper sulfate
37

450.00
400.00
Total Dissolved Solids 350.00
300.00
250.00
200.00
150.00
100.00
50.00
0.00
0.2g 0.5g 0.8g
Table 5 shows the Analysis of Variance for copper sulfate with fifteen (15) minutes
Turbidity has a significance of 0.279 which is also non-significant. Total Dissolved
Solids has a significance value of 0.572 which is also non-significant.
38
ANOVA

pH Between Groups .022 .011 2.622 .152
Within Groups .025 .004
Total .048
Turbidity Between Groups .730 .365 1.592 .279
Within Groups 1.376 .229
Total 2.106
TDS Between Groups 2605.556 1302.778 .615 .572
Within Groups 12720.000 2120.000
Total 15325.556
Table 6 shows the correlations for copper sulfate with 15 minutes contact time.
The significance level is set to p ≤ 0.01, value lower or exact to 0.01 implies rejection of
null hypothesis and value higher than 0.01 implies that results failed to reject null
hypothesis. Only between pH level and TDS obtained a p-value of 0.008 which is lower
than the alpha value of 0.01, implicating that it has a significant relationship. The rest
showed insignificant values such as: 0.885 between TDS and turbidity; and 0.405
between pH and turbidity.
Correlations
pH Turbidity TDS
pH Pearson Correlation 1 .317 -.812**
Sig. (2-tailed) .405 .008
N 9 9 9
Turbidity Pearson Correlation .317 1 .056
39
Sig. (2-tailed) .405 .885

N 9 9 9
**
TDS Pearson Correlation -.812 .056 1
Sig. (2-tailed) .008 .885
N 9 9 9
acidic and value greater than 7 is considered basic. The determination of pH Level used
Muntingia calabura exhibited a mean of 4.38 as the highest, 0.5g of powdered Muntingia
calabura has a mean of 4.28 and 0.8g has a mean of 4.21as the lowest. Then, as the
biosorbent dose increases the pH level decreases with 25 minutes contact time.
for Copper sulfate

40

4.70
4.60
4.50
4.40
pH Level
4.30
4.20
4.10
4.00
3.90
0.2g 0.5g 0.8g
time on Turbidity per trial. The results were determined with the use of turbidimeter. The
sample with 0.2g biosorbent dose had a mean result of 0.35 as the highest while the
second sample with 0.5g obtained 0.20 mean which was the lowest and the samples
containing 0.8g garnered same 0.35 mean as samples with 0.2g.
Figure 8. Effect of Varying Biosorbent Dose with 25 minutes Contact on
Turbidityfor Copper sulfate

41

0.70
0.60
0.50
0.40
Turbidity
0.30
0.20
0.10
0.00
0.2g 0.5g 0.8g
time on TDS (Total Dissolved Solids). The results were measured through the use of
Electrical Conductivity (EC) meter. There was an increased amount of TDS from the
sample containing 0.2g of biosorbent dose which had a mean of 299.67, 0.5g sample
obtained 339.67 mean and 0.8g had the highest result of 354.00.
Copper sulfate
42

500.00
450.00
Total Dissolved Solids 400.00
350.00
300.00
250.00
200.00
150.00
100.00
50.00
0.00
0.2g 0.5g 0.8g
Table 7 shows the Analysis of Variance for copper sulfate with Twenty-five (25)
minutes contact time. The significance level is set to p ≤ 0.05, value lower or exact to
0.05 implies rejection of null hypothesis and value higher than 0.05 implies that results
failed to reject null hypothesis. The pH level has a significance of 0.426 which is non-
significant. Turbidity has a significance of 0.511 which is also nonsignificant. Total
Dissolved Solids has a significance value of 0.878 which is also non-significant.
43
ANOVA
Sum of
Squares Mean Square F Sig.

pH Between Groups .042 .021 .986 .426
Total .169
Turbidity Between Groups .048 .024 .751 .511
Total .240
Within Groups 107111.333 17851.889
Total 111868.889
Table 8 shows the correlations for copper sulfate with 25 minutes contact time.
The significance level is set to p ≤ 0.01, value lower or exact to 0.01 implies rejection of
null hypothesis and value higher than 0.01 implies that results failed to reject null
hypothesis. Only between pH level and TDS obtained a p-value of 0.001 which is lower
showed insignificant values such as: 0.669 between TDS and turbidity; and 0.942
between pH and turbidity.
Correlations
pH Turbidity TDS
pH Pearson Correlation 1 -.028 -.915**
Sig. (2-tailed) .942 .001
N 9 9 9
Turbidity Pearson Correlation -.028 1 -.166
44
Sig. (2-tailed) .942 .669

N 9 9 9
TDS Pearson Correlation -.915** -.166 1
Sig. (2-tailed) .001 .669
N 9 9 9
time on pH Level per trial. The neutral pH of water is 7, value lesser than 7, is considered
Muntingia calabura exhibited a mean of 4.87 as the lowest, 0.5g of powdered Muntingia
calabura has a mean of 5.13 and 0.8g has a mean of 5.15as the highest and as the
biosorbent dose increases the pH level also increases.
for Zinc sulfate

45
Zinc sulfate 5mins

5.30
5.20
5.10
5.00
pH Level
4.90
4.80
4.70
4.60
4.50
4.40
0.2g 0.5g 0.8g
time on Turbidity for Zinc Sulfate per trial. The results were determined with the use of
turbidimeter. The sample with 0.2g biosorbent dose had a mean result of 2.63 as the
highest while the second sample with 0.5g obtained 1.61 mean which was the lowest and
the trials containing 0.8g garnered 1.76.
for Zinc sulfate

46
Zinc sulfate 5mins

4.50
4.00
3.50
3.00
Turbidity
2.50
2.00
1.50
1.00
0.50
0.00
0.2g 0.5g 0.8g
of Electrical Conductivity (EC) meter. There was a decreased in amount of TDS from the
sample containing 0.2g of biosorbent dose which had a mean of 261.63 to 207.67 mean
obtained from 0.5g then increased in amount of 293.67 mean at 0.8g sample.
Zinc sulfate
47
Zinc sulfate 5mins

450.00
400.00
350.00
300.00
250.00
200.00
150.00
100.00
50.00
0.00
0.2g 0.5g 0.8g
Table 9 shows the Analysis of Variance for zinc sulfate with Five (5) minutes contact
time. The significance level is set to p ≤ 0.05, value lower or exact to 0.05 implies
has a significance value of 0. 710 which is also non-significant.
48
ANOVA
pH Between Groups .143 .071 4.910 .055

Total .230
Turbidity Between Groups 1.820 .910 .461 .651
Within Groups 11.847 1.974
Total 13.667
Within Groups 93800.940 15633.490
Total 105135.476
Table 10 shows the correlations for zinc sulfate with 5 minutes contact time. The
Only between pH level and TDS obtained a p-value of 0.127 which is higher than the
alpha value of 0.01, having an insignificant relationship. The rest showed significant
values such as: 0.005 between TDS and turbidity; and 0.013 between pH and turbidity.
Correlations
pH Turbidity TDS
pH Pearson Correlation 1 -.782* -.548
Sig. (2-tailed) .013 .127
N 9 9 9
Turbidity Pearson Correlation -.782* 1 .836**
Sig. (2-tailed) .013 .005
49
N 9 9 9
TDS Pearson Correlation -.548 .836** 1
Sig. (2-tailed) .127 .005
N 9 9 9
*. Correlation is significant at the 0.05 level (2-tailed).
contact time on pH Level per trial. The neutral pH of water is 7, value lesser than 7, is
considered acidic and value greater than 7 is considered basic. The determination of pH
Level used Electrometric method through pH meter device. Samples with 0.2g of
powdered Muntingia calabura exhibited a mean of 5.22 which is the same with the mean
pH level value of 0.8g, while 0.5g of powdered Muntingia calabura has a mean of 5.21as
the lowest. Thus from the 0.2g biosorbent dose, the pH level decreased at 0.5g dose and it
increased back again to the first mean value at 0.8g dose.
for Zinc sulfate

50
Zinc sulfate 15mins

5.35
5.30
5.25
5.20
pH Level
5.15
5.10
5.05
5.00
0.2g 0.5g 0.8g
time on Turbidity for Zinc Sulfate per trial. The results were determined with the use of
turbidimeter. The sample with 0.2g biosorbent dose had a mean result of 0.84 as the
highest while the second sample with 0.5g obtained 0.99 mean and the samples
containing 0.8g garnered 0.62 which was the lowest.
for Zinc sulfate

51
Zinc sulfate 15mins

2.50
2.00
1.50
Turbidity
1.00
0.50
0.00
0.2g 0.5g 0.8g
of Electrical Conductivity (EC) meter. There was an increased amount of TDS from the
sample containing 0.2g biosorbent dose which had a mean of 224.33 to the 267.33 result
obtained from 0.5g sample then decreased in amount of 224.67 at 0.8g dose.
Zinc sulfate
52
Zinc sulfate 15mins

400.00
350.00
300.00
250.00
200.00
150.00
100.00
50.00
0.00
0.2g 0.5g 0.8g
Table 11 shows the Analysis of Variance for zinc sulfate with Fifteen (15) minutes
has a significance value of 0.788 which is also non-significant.
53
ANOVA
Total .029
Turbidity Between Groups .207 .103 .257 .782
Within Groups 2.420 .403
Total 2.627
Within Groups 44366.000 7394.333
Total 48035.556
Only between turbidity and TDS obtained a p-value of 0.623 which is higher than the
alpha value of 0.01, having an insignificant relationship. The rest showed significant
values such as: 0.034 between pH level and turbidity; and 0.002 between pH and TDS.
Table 12. Correlations Between pH, Turbidity and TDS of Copper sulfate
with 15 minutes Contact Time
Correlations
pH Turbidity TDS
pH Pearson Correlation 1 -.705* -.881**
Sig. (2-tailed) .034 .002
N 9 9 9
*
Turbidity Pearson Correlation -.705 1 .623
Sig. (2-tailed) .034 .073
54
pH Turbidity TDS
pH Pearson Correlation 1 -.705* -.881**
N 9 9 9
TDS Pearson Correlation -.881** .623 1
Sig. (2-tailed) .002 .073
N 9 9 9
Muntingia calaburaexhibited a mean of 5.17, the value then decreased at 0.5g of
powdered Muntingia calabura with a mean of 5.16as the lowest. Then at 0.8g biosorbent
dose it obtained a mean of 5.19as the highest pH level mean value.
for Zinc sulfate

55
Zinc sulfate 25mins

5.24
5.22
5.20
5.18
pH Level
5.16
5.14
5.12
5.10
5.08
0.2g 0.5g 0.8g
contact time on Turbidity for Zinc sulfate per trial. The results were determined with the
use of turbidimeter. The sample with 0.2g biosorbent dose had a mean result of 1.66 as
the highest while the second sample with 0.5g obtained 0.37 mean and the samples
containing 0.8g garnered 0.21 which was the lowest.
for Zinc sulfate

56
Zinc sulfate 25mins

2.50
2.00
1.50
Turbidity
1.00
0.50
0.00
0.2g 0.5g 0.8g
The Figure 18 below, shows the effect of varying biosorbent dose with 25 minutes
of Electrical Conductivity (EC) meter. There was a decreased amount of TDS from the
sample containing 0.2g biosorbent dose which had a mean of 324.33, 0.5g sample
obtained 267.33 result then 0.8g sample had the lowest result of 224.67.
Zinc sulfate
57
Zinc sulfate 25mins

400.00
Total Dissolved Solids(Mean) 350.00
300.00
250.00
200.00
150.00
100.00
50.00
0.00
0.2g 0.5g 0.8g
Table 13 shows the Analysis of Variance for zinc sulfate with Twenty-five (25)
minutes contact time. The significance level is set to p ≤ 0.05, value lower or exact to
0.05 implies rejection of null hypothesis and value higher than 0.05 implies that results
failed to reject null hypothesis. The pH level has a significance of 0.517 which is non-
significant. Turbidity has a significance of 0.003 which is significant. Total Dissolved
Solids has a significance value of 0.061 which is also non-significant.
58
ANOVA
Sum of
Squares Mean Square F Sig.

Total .008
Turbidity Between Groups 3.818 1.909 16.946 .003
Total 4.494
TDS Between Groups 10204.667 5102.333 4.618 .061
Within Groups 6629.333 1104.889
Total 16834.000
Table 14 shows the Multiple Comparisons for Turbidity by Zinc sulfate with
Twenty-five (25) minutes contact time. Tukey HSD was used (description) and illustrated
due to the significant value exhibited during the analysis of variance. Samples represent
the different treatments with varying dosages such that: sample 1 is contains 0.2g of
biosorbent dose; sample 2 contains 0.5g dosage; and sample 3 has 0.8g amount of dose.
Between sample 1 and sample 2, a p-value of 0.008 was shown, which is lower than the
alpha value of 0.05 a significant value of 0.004 was also obtained between sample 1 and
sample 3 which is also lower than the alpha value. Then, the significance value between
samples 2 and 3 garnered 0.840 which is higher than the alpha value of 0.05. Thus,
between samples 1 and 2 then on samples 1 and 3, showed significant differences while
between samples 2 and 3 their p-value implies that there is no significant difference.
Table 14. Multiple Comparisons for Turbidity

59
Multiple Comparisons (Tukey HSD)
Dependent (I) (J) Mean Difference
Variable Sample Sample (I-J) Std. Error Sig.

Turbidity 1.00 2.00 1.29667* .27405 .008
3.00 1.45333* .27405 .004
2.00 1.00 -1.29667* .27405 .008
3.00 .15667 .27405 .840
*
3.00 1.00 -1.45333 .27405 .004
2.00 -.15667 .27405 .840
Between turbidity and TDS obtained a p-value of 0.032 which is lower than the alpha
value of 0.05, implicating that it has a significant relationship. The rest showed
insignificant values such as: 0.209 between TDS and pH; and 0.854 between pH and
turbidity.
Correlations
pH Turbidity TDS
pH Pearson Correlation 1 -.072 -.464
Sig. (2-tailed) .854 .209
N 9 9 9
Turbidity Pearson Correlation -.072 1 .711*
Sig. (2-tailed) .854 .032
N 9 9 9
TDS Pearson Correlation -.464 .711* 1
60
Sig. (2-tailed) .209 .032

N 9 9 9
Figure 19 below, shows the pH Level means in varying contact time and varying
biosorbent dose. For 5min and 15min samples with 0.2g of powdered Muntingia
calabura exhibited values of 4.23 and 4.19 mean. The values then increased with ones
having 0.5g, which are4.45 and 4.29 and they again increased with samples containing
0.8g, with values of 4.47 and 4.30. Hence, 5 and 15 min contact time showed inclining
result as the biosorbent dose increases. As for 25 minutes contact time from 0.2g
biosorbent dose with 4.38 mean value, it decreased to 4.28 when the dose was increased
to 0.5g, and then it again decreased to 4.21 with 0.8g of biosorbent dose which showed
declining results as biosorbent dose increases. Thus, 15min of contact time with 0.2g of
biosorbent dose obtained the lowest pH level mean values while the highest was
exhibited during 5min contact time with 0.8g biosorbent dose and as the contact time
increases the pH level rises.
Time for Copper sulfate

61
Copper sulfate
4.50
4.45
pH Level (Mean) 4.40

4.35
4.30
4.25
4.20
4.15
4.10
4.05
4.00
0.2g 0.5g 0.8g
Figure 20 below, shows the turbidity means for varying contact time and
biosorbent dose.Varied biosorbent doses with 5 minutes contact time shows decrease
from 0.64 to 0.36 and increase to 0.55.Treatments subjected to 15 minutes contact time
also showed decrease from 0.50 to 0.33 and increase to 1.00, exhibiting the highest mean.
Treatments with 25 minutes time exhibited decrease from 0.35 to 0.20 being the lowest
mean and increase to 0.35. The comparison of varied biosorbent dose shows declining
trends from 0.2g treatments to 0.5g treatment and increasing trend with 0.8g treatments.
It was observed in Figure 20 that copper sulfate absorption rate showed best results with
treatments with 0.5g of powdered Muntingia calabura leaves subjected to 25 minutes
contact time.
62
Copper sulfate
1.20
1.00
Turbidity (Mean)
0.80
0.60
0.40
0.20
0.00
0.2g 0.5g 0.8g
Figure 21 below, shows the effect of biosorbent dose with varying contact time on
TDS (Total Dissolved Solids). There was a decreased in amount of TDS on the varied
absorbent doses with 5minutes contact time from 361.33 to 281.93 and lowest mean of
63
255.27. Treatments with 15 minutes showed a decrease mean from 380.67 to 342.33
being the lowest and increase to 347.33. Treatments with 25 minutes showed an inclining
result obtained from the first sample which had a mean of 299.67 to 339.67 and to the last
sample which had the highest mean of 354.00. Thus, treatments with 5 minutes had a
declining results from 0.2g to 0.8g, treatments with 15 minutes had a decreasing result
from 0.2g to 0.5g and increase at 0.8g and treatments with 25 minutes had inclining
results from 0.2g to 0.8g. It was observed in Figure 21 that copper sulfate biosorption rate
showed effective results with treatments with 0.8g of powdered Muntingia calabura
leaves subjected to 5 minutes contact time.

64
Figure 21. TDS Level Mean Comparisons with Varying Biosorbent Dose and
Copper sulfate
400.00
Total Dissolved Solids (Mean)
350.00
300.00
250.00
200.00
150.00
100.00
50.00
0.00
0.2g 0.5g 0.8g
Figure 22 below, shows thepH level mean comparisons with varying biosorbent
dose and contact time for Zinc sulfate. For 15min and 25min samples with 0.2g of
powdered Muntingia calabura exhibited values of 5.22 and 5.17 mean. The values then
65
decreased with ones having 0.5g, which are 5.21 and 5.16, and then they increased with
samples containing 0.8g, with values of 5.22 and 5.19. As for 5 minutes contact time
from 0.2g biosorbent dose with 4.87mean value, it increased to 5.13 when the dose was
increased to 0.5g, and then it again increased to 5.15 with 0.8g of biosorbent dose which
showed inclining results as biosorbent dose rises. Thus, 5min of contact time with 0.2g of
biosorbent dose obtained the lowest pH level mean values while the highest was
exhibited during 15minutes contact time with 0.2g and 0.8g biosorbent dose. It was
observed in Figure 22 that biosorption rate of copper sulfate showed effective results with
15 minutes contact time in varying biosorbent doses and it also exhibited optimum results
with 0.8 biosorbent dose in varying contact time.
Figure
Zinc sulfate
5.30
22. PH
5.20
pH Level (Mean)
5.10 Level
5.00
4.90
Mean
4.80
4.70
4.60
0.2g 0.5g 0.8g
Comparisons with Varying Biosorbent Dose and Contact Time for Zinc sulfate
66
Figure 23 below, shows the turbidity means for varying contact time and
biosorbent dose. Varied biosorbent doses with 5 minutes contact time shows decrease
from 2.63 to 1.61 and increase to 1.76 in which the treatment exposed to 0.2g biosorbent
dose exhibited the highest mean. Treatments subjected to 15 minutes contact time
showed increase from 0.84 to 0.99 and decreases to 0.62. Treatments with 25 minutes
time exhibited decrease from 1.66 to 0.37 and decrease to 0.21, being the lowest. The
comparison of varied biosorbent dose shows declining trends from 0.2g treatments to
0.5g treatments with 5min and 15min contact time and they again decreased with 0.8g
treatments with 15min and 25 min contact time. It was observed in Figure 23 that Copper
sulfate absorption rate showed best results with treatments with 0.8g of powdered
Muntingia calabura leaves subjected to 25 minutes contact time.

67
Figure 23.Turbidity Level Mean Comparisons with Varying Biosorbent Dose and
Zinc sulfate
3.00
2.50
2.00
Turbidity (Mean)
1.50
1.00
0.50
0.00
0.2g 0.5g 0.8g
Figure 24 below, shows the effect of biosorbent dose with varying contact time on
TDS (Total Dissolved Solids). There was a decreased in amount of TDS on the varied
biosorbent doses with 5minutes contact time from 261.63 to 207.67 and increased to
68
293.67. Treatments with 15 minutes showed an increasing mean from 224.33 to 267.33
being the highest and decrease to 224.67. Treatments with 25 minutes showed a declining
results obtained from the first sample which had a mean of 324.33 to 303.00 and to the
last sample which had the lowest mean of 244.07. Thus, treatments with 5 minutes had a
declining results from 0.2g to 0.5g and incline at 0.8g, treatments with 15 minutes had an
increasing result from 0.2g to 0.5g and decrease at 0.8g and treatments with 25 minutes
had declining results from 0.2g to 0.8g. It was observed in Figure 24 that zinc sulfate
biosorption rate showed effective results with treatments with 0.5g of powdered
Muntingia calabura leaves subjected to 5 minutes contact time.

69
Figure 24. TDS Level Mean Comparisons with Varying Biosorbent Dose and
Zinc sulfate
350.00
300.00
Total Dissolved Solids (Mean)
250.00
200.00
150.00
100.00
50.00
0.00
0.2g 0.5g 0.8g
Summary of Findings
Experimental studies from past researches showed that biosorption is dependent
on different time intervals. Study agrees with previous experiments showing that
biosorption takes place phases such that initial contact time of 25 min made rapid surface
70
biosorption and it becomes slow after succeeding time with its biosorption rate in which
then (Rao et. al, 2003 as cited in Reena, Malik and Singhal, 2015, p. 11, par. 2). Jena and
Sahoo (2017, p. 7, par. 4) adds that the equilibrium was reached within 40 minutes and
thereafter the rate of metal removal remained almost stable with a slower rate. A
significant number of vacant active binding sites was available at the first phase of
experiment but the binding site becomes limited and the remaining vacant surface sites
were difficult to be occupied by metal due to formation of repulsive forces. Hence,
Muntingia calaburaleaves during experimentations exhibited lowest TDS values during
5 min of biosorbent contact time for both Copper and Zinc solutions.
In line with the data of Jena et.al (2017, table 4) with increasing biosorbent dose,
shows optimum percentage of inhibition during the increase of biosorbent concentration.
They added that it is attributed due to more binding sites or increasing surface area with
free ions remaining constant for higher chemical biosorption. Additionally, the Total
Dissolved Solids (TDS) values of copper and zinc solutions also decrease as the
Muntingia calaburapowder increases and mostly efficient during 0.8g dosage as
compared to 0.2g and 0.5g doses.
In proportion to the data of Geoffrey et.al, (2018, Fig.1) article no. CSIJ 39672
(Chemical Science International Journal), the effect of turbidity and contact time on the
biosorption of the heavy metals by the Moringaoleiferabiosorbent showed that increasing
of biosorbent dose, the percentage of sorption of the metal ions decreased with increase
in turbidity. In this case, the turbidity values of copper solutions at 5 minutes contact time
and all of the zinc solutions on varying contact time also decrease as the
71
Muntingia calaburapowder increases and is mostly effective during 0.8g dosage at 25
minutes contact time as compared to 0.2g and 0.5g doses with 5 and 15 minutes.
Based on the results of Salezadeh (2013, p. 101, table 1), the removal percentage
was observed on between different heavy metals, which are commonly hazardous and
abundant. During the experimentation for the increase of adsorbent dose, not only that the
increase in adsorbent dose showed increasing percentage removal, it also showed that
higher adsorption performance was shown for copper than as compared to zinc. The
study also supports the current results of the researchers using Muntingiacalaburaon
copper sulfate and zinc sulfate, for results showed more significant biosorption
performance on copper than on zinc solutions.
The researchers found that as the biosorbent dose increases the pH level also
increases both on the copper and zinc solutions. The pH level and TDS, showed
significant relationships where it implicates that as the biosorbent dose increases, the pH
level increases and the TDS levels on the other hand decreases in value. There was also a
significant relationship obtained between pH level and turbidity in which as the
biosorbent dose increases, pH increases and turbidity decreases.
In addition to this, on the correlation between the pH, Turbidity, and TDS of
copper sulfate at 5, 15, and 25 minutes contact time, only between pH level and TDS
obtained a p-value of 0.001at 5 and 25 minutes and 0.008 at 15 minutes which is lower
showed insignificant values such as between TDS and turbidity. However, between pH
and turbidity on the zinc sulfate, the correlation between each parameters showed
significant values at 15 minutes contact time such as: 0.034 between pH level and
72
turbidity, 0.002 between pH and TDS at 25 minutes and 0.013 between pH and turbidity
at 5 minutes contact time implicating that it has a significant relationship. The rest
showed insignificant values between pH and turbidity with 25 mins and between pH and
TDS at 5 minutes contact time. Hence, it showed no significant difference in varying
biosorbent doses and contact time on almost all the prepared treatments, except for the
turbidity of Zinc sulfate exposed to 25 min in terms of varying biosorbent dose.
Thus, positive potential implications was inferred based on the descriptive data,
due to the optimum results exhibited on the highest dosage of biosorbentin terms of Total
Dissolved Solids and pH level of the solution. As for turbidity, better results were mostly
found with 0.5g dosage because less amount of dose, leaves copper sulfate and zinc
sulfate levels high. Although,too much biosorbent dosage exhibits higher removal
capacity, it leaves biomass stains on the solution. This observation was supported by a
review entitled State of the Art Biosoprtion Process, it stated that one of the major
disadvantages of biosorption is the difficulty of separating the sorbent from the solution.
Conclusion
Muntingia calabura Linn (Sarisa) powder proved to be a potential biosorbent on
Copper sulfate and Zinc sulfate aqueous solutions in terms on its effects on the significant
73
relationships between pH level with Total Dissolved Solids and its proportion on
turbidity. It also showed potential to be an eco-friendly and abundant source of
biosorbent for certain chemical contaminations, though it showed no significant
difference on the variation of biosorbent doses.
The first null hypothesis stating that there is no significant difference
difference in the Copper sulfate and Zinc sulfate biosorbent potential of powdered
Muntingia calabura Linn (Sarisa)leaves in aqueous solutions prepared in different
biosorbent dose and contact time and tested in terms of pH level, turbidity, and TDS
(total dissolved solids) is accepted.
Based on the results between pH level and Total Dissolved Solids the
second null hypothesis stating that there is no significant relationship between the varying
biosorbent dose and contact time of Powdered Muntingia calabura Linn (Sarisa) leaves
and its biosorbing potential of Copper sulfate and Zinc sulfate in terms of pH level,
turbidity and Total Dissolved Solids is rejected since it showed significant relationships.
In conclusion, Muntingia calabura is a low-cost biosorbent, exhibiting low biosorptive
potential against copper sulfate and zinc sulfate from aqueous solutions.
74
Recommendations
The researchers observed fluctuating results during the trials thus it is
recommended that the trials should be increased to 5-20 trials in order to get more
accurate and legitimate results. Researchers also wanted to include more parameters such
as: effect of chemical or biosorbate concentration; effect of temperature; and effect of pH
level and work on the experiments with optimum conditions. The use of much more
precise devices and additional materials: such as a rotary shaker than using a centrifuge;
mixing gel beads to the biosorbents for more efficient distribution in the solution;
vacuum analytical balance; Fourier Transform Infrared Spectroscopy for scanning test
samples and chemical properties; and Microfibrous Entrapped Sorbent (MFES)
Technology; is very much encouraged for next researches. It is also highly recommended
to make and characterize an activated carbon using Muntingia calabura Linn (Sarisa)
leaves for heavy metal adsorption. More studies are also suggested using different
chemicals like Arsenic, Cadmium, Manganese and Molybdenum which are also
alarmingly hazardous to water supplies and agricultural land areas within and out of the
country.
75
Acknowledgment
The researchers would like to thank all the people who took part in the study. First
are all the teachers who supported them, especially the following: Mr. Nathan Fernandez
our co-adviser in research, for giving the primary lessons that was very crucial before
beginning the research study and for being a very supportive teacher; Ms. Mercuria
Recto, the laboratory head Bacolod City National High School for guiding and providing
the materials needed during the experimentation; Mr. Villaruz for helping to solve the
research’s statistical results and analysis; and last but not the least, Mr. Phillip Raymund
De Oca for supporting the researchers in many aspects and for almost being there every
step of the way. The help Mrs. Arlene Dela Rama as a qualified scientist for verifying the
steps and the study itself is also highly recognized. Mr. John A. Calumpiano for the plant
verification, Mr. Churchill Lacuesta for the moral support and other personnel from the
Bureau of Plants Industry Quarantine Services are highly appreciated. Laboratory
technicians from San Agustin College, Mr. Xerxes Hinolan and his co-staff are very
much thanked for providing crucial help during the experimentation of the study. People
from BACIWA laboratory especially Ms. April Rose P. Regalado, Ms. Gladys M.
Taisipit and Ms. Daisy V. Desengano for providing assistance to make the analysis of
results for the study, possible. The researchers would like to thank also their parents and
guardians who supported by providing their needs such as: financial, mental and
emotional support. They also thank all their classmates and friends for the undying moral
support. Lastly, the researchers would like to praise and thank the Lord for the love and
blessings of knowledge, wisdom and strength to do their best and to finish the study.
76
References
Aathithya, R., Rajani J. Sowparnika, and V. Balakrishnan. 2014. "Determination of
adsorption isotherm models for the biosorption of chromium using cherry leaves
(Muntingia calabura L.)." Accessed December 2018.Arvanitoyannis, Ioannis S.,
and Theodoros H. Varzakas. Activated Carbon. 2008.
https://www.sciencedirect.com/topics/agricultural-and-biological-
sciences/activated-carbon (accessed September 12, 2018).
Department of Health. 2010.
http://www.health.gov.au/internet/publications/publishing.nsf/Content/ohp-
enhealth-manual-atsi-cnt-l~ohp-enhealth-manual-atsi-cnt-l-ch6~ohp-enhealth-
manual-atsi-cnt-l-ch6.1.
Geoffrey, Chavaregi, J. L. Kituyi, and S. Lutta. Chemical Science International Journal.
June 18, 2018.
http://www.journalrepository.org/media/journals/CSIJ_53/2018/Jun/Geoffrey232
2018CSIJ39672.pdf (accessed February 2019).
Jena, Swarnabala, and Rajesh Kumar Sahoo. “Removal Of Pb(Ii) From Aqueous Solution
Using Fruits. 2017.
https://pdfs.semanticscholar.org/f7c3/06d9bb510725c7cabaa0819ca6f2556fa7cb.
pdf (accessed September 2018).
Lenntech. n.d. https://www.lenntech.com/processes/heavy/heavy-metals/heavy-
metals.htm (accessed 2019).

77
Marcotullio, Peter J. "Urban water-related environmental transition in southeast asia."
Sustainability Science (Springer Japan) 2, no. 1 (April 2007): 27-54.
Michalak, Izabela, Katarzyna Chojnacka, and Anna Witek-Krowiak. State of the Art for
the Biosorption Process—a Review. May 12, 2013.
https://link.springer.com/article/10.1007/s12010-013-0269-0 (accessed February
2019).
N. D. Mahmood, N. L. M. Nasir, M. S. Rofiee, S. F. M. Tohid, S. M. Ching, L. K. 2014.
Muntingia calabura: A review of its traditional uses,chemical properties, and
pharmacological observations,.
Salehzadeh, Jaber. Leonardo Journal of Sciences. December 2013.
http://ljs.academicdirect.org/A23/097_104.pdf (accessed February 2019).
Santhi, T. , S. Manonmani, and S. Ravi. 2009. "Uptake of Cationic Dyes from Aqueous
Solution by." E-Journal of Chemistry 737-742.
Suhadiyah, Sri, Djamal Sanusi, Samuel Paembonan, and Roland A. Barkey.ijstr.org.
March 2013. http://www.ijstr.org/final-print/mar2013/Lead-Accummulation-
Potential-By-Leaves-With-Abundant--Trichomes-Muntingia-Calabura-L.-And-
Rare-Trichomes-Mimusops-Elengi-L.-In-Makassar-Indonesia.pdf (accessed
September 1, 2018).
Thankur, Lokendra Singh. researchgate.net. 2013.
https://www.researchgate.net/publication/271281628_Heavy_metal_Cu_Ni_and_
Zn_Toxicity_health_hazards_and_their_removal_techniques_by_low_cost_adsor
bents_A_short_overview.
78
Appendices
Cost Analysis
30L Distilled Water Php 350
(45) Test Tubes Php 600
(42) Bottle Containers Php 50
Statistician Php 500
Laboratory Fees Php 600
Miscellaneous Php 400
Php 2500
79
Documentation
80
Figure 25. Gathering and Figure 26. Washing of Figure 27. Weighing of
weighing of Muntingia Muntingia calabura leaves washed Muntingia calabura
calabura leaves Leaves
Figure 28. Drying of Figure 29. Weighing of Figure 30. Pulverizing

Muntingia calabura leaves dried Muntingia calabura Muntingia calabura leaves
leaves
Figure 31. Muntingia Figure 32. Oven-drying of Figure 33. Weighing of oven-
calabura leaves in powdered Muntingiac alabura dried powdered Muntingia
powdered form leaves calabura leaves
81
Figure 34. Weighing of 0.2 g of Figure 35. Weighing of 0.5g Figure 36. Weighing of 0.8g
powdered Muntingia calabura of powdered Muntingia of powdered Muntingia
leaves calabura leaves calabura leaves
Figure 39. Distilled water

82
Figure 37. Weighing of 30g Figure 38. Weighing of 30g

Copper sulfate Zinc sulfate
Figure 40. Mixing of 300mL Figure 41. Mixing of 300mL Figure 42. Gathering of
Copper sulfate solution Zinc sulfate solution Materials and Equipment
Figure 43. Preparation of samples Figure 44. Centrifugation of Figure 45. Placing sample
prepared samples solutions on the test tube racks
83
Figure 46. Filtering Samples Figure 47. Transferring samples Figure 48. Analyzing water of
to bottle containers samples and recording results
84
Curriculum Vitae
Name: Maria Isabel T. Garcia
Address: Ilang-ilang St., Brgy. Mansilignan, Bacolod City,Philippines
Age: 16
Civil Status: Single
Father’s Name: Glen Peter G. Garcia
Occupation: College Math Teacher
Mother’s Name: Jizza T. Garcia
Occupation: Homemaker
Educational Attainment:
Elementary: Antonio L. Jayme Elementary School
High School: Bacolod City National High School
Honor/Awards/Scholarship:
2014-2015: 1st Honorable mention
2015-2016: Math Awardee (Gold & Silver), Science Awardee (Silver)
2016-2017: With Honors, Math Awardee (Silver), Research Awardee (Silver)
2017-2018: With Honors
Course(s) to be taken: BS Mathematics

85
Name: Ashlie Jane B. Balido
Address: Blk.7 Lot 9, NEDF Village, Brgy. Handumanan, Bacolod City, Philippines
Age: 16
Father’s Name: Agustin T. Balidio
Occupation: Computer Technician
Mother’s Name: Jocelyn B. Balidio
Occupation: Housewife
Elementary: Handumanan Integrated Christian Academy
2014-2015: Valedictorian
2015-2016: 7th honor Science Awardee (Gold), Math Awardee (Gold)
2016-2017: With honors, Science Awardee (Silver), Research Awardee (Silver)
2017-2018: With high honors
Course(s) to be taken: Engineering

86
Name: JB Shiela I. Munieza
Address: Blk. 5, Lot 9, Garces-Garcia Village,Purok Paglaum,Brgy. Taculing,Bacolod
City,Philippines
Age: 15
Father’s Name: Bernardino T. Munieza
Occupation: Technician
Mother’s Name: Janice V. Isip
Occupation: Factory Worker
Elementary: Don Crispino V. Ramos Elementary School
2014-2015: 5th Honor
2015-2016:
2016-2017
2017-2018
Course(s) to be taken:
87
Letters

Muntingia Calabura

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1

Characterization of Powdered Muntingia calabura L. (Sarisa) Leaves as Potential

ASHLIE JANE B. BALIDIO

MARIA ISABEL T. GARCIA

PHILLIP RAYMUND DE OCA

BACOLOD CITY NATIONAL HIGH SCHOOL

Bacolod City, Negros Occidental

Region VI, Western Visayas

water-intensive goods continually rise. Heavy metals are characterized as natural

Muntingia calabura Linn (Sarisa) leaves in aqueous solutions prepared in different

different contact times of 5, 15 and 25 minutes for each chemical solution.Between

as time was added. In conclusion, Muntingiacalabura is a low-cost biosorbentexhibiting

Official Abstract Form 2

Results and Discussion 27

Appendices Cost Analysis 77

Figure 2. Effect of Varying Biosorbent Dose with 5 minutes Contact on Turbidity

for Copper sulfate

Figure 4. Effect of Varying Biosorbent Dose with 15 minutes Contact on pH Level

for Copper sulfate

Figure 5. Effect of Varying Biosorbent Dose with 15 minutes Contact on Turbidity

for Copper sulfate

Figure 7. Effect of Varying Biosorbent Dose with 25 minutes Contact on pH Level

for Copper sulfate

Figure 8. Effect of Varying Biosorbent Dose with 25 minutes Contact on Turbidity

for Copper sulfate

for Zinc sulfate

for Zinc sulfate

for Zinc sulfate

for Zinc sulfate

for Zinc sulfate

for Zinc sulfate

Time for Copper sulfate

Contact Time for Copper sulfate

Contact Time for Copper sulfate

Time for Zinc sulfate

Contact Time for Zinc sulfate

Contact Time for Zinc sulfate

Figure 25. Gathering and weighing of Muntingia calabura leaves

Figure 26. Washing of Muntingia calabura leaves

Figure 27. Weighing of washed Muntingia calabura Leaves

Figure 28. Drying of Muntingia calabura leaves

Figure 29. Weighing of dried Muntingia calabura leaves

Figure 30. Pulverizing Muntingia calabura leaves

Figure 31. Muntingia calabura leaves in powdered form

Figure 32. Oven-drying of powdered Muntingia calabura leaves

Figure 33. Weighing of oven-dried powdered Muntingia calabura leaves

Figure 34. Weighing of 0.2 g of powdered Muntingia calaburaleaves

Figure 35. Weighing of 0.5g of powdered Muntingia calabura leaves

Figure 36. Weighing of 0.8g of powdered Muntingia calabura leaves

Figure 37. Weighing of 30g Copper sulfate

Figure 38. Weighing of 30g Zinc sulfate

Figure 39. Distilled water

Figure 40. Mixing of 300mL Copper sulfate solution

Figure 41. Mixing of 300mL Zinc sulfate solution

Figure 42. Gathering of Materials and Equipment

Figure 43. Preparation of samples

Figure 44. Centrifugation of prepared samples

Figure 45. Placing sample solutions on the test tube racks

Figure 46. Filtering Samples

Figure 47. Transferring samples to bottle containers

Figure 48. Analyzing water of samples and recording results

on Copper sulfate in Treatments A, B, and C

on Zinc sulfate in Treatments A, B, and C