(Final Manuscript) BIo-Piezoelectric Nanogenerator - Harnessing Electrical Energy From The Fishbones of Tilapia (Oreochromis Mossambicus)

Senior High School Department
Laguna Campus
Bio-Piezoelectric Nano generator: Harnessing Electrical Energy
From Fish Bones of Tilapia (Oreochromis mossambicus)
Baliwag, Jesse Lambert R.
Brazil, Gian Miguel C.
Perez, Gabriel Francis D.
Rufino, Arvin Gabriel D.
Sambrano, Alphonse Rossaint V.
3rd Trimester, A.Y. 2018-2019
April 10, 2019

2019. All Rights Reserved. No part of this publication may be reproduced, stored in an
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mechanical, by photocopying, scanning, recording, or otherwise, except under the terms of the
applicable law.
Running Head: BIO-PIEZOELECTRIC NANO GENERATOR i
Acknowledgement
We, the researchers would like to thank a handful of people for their generous
contributions that has led to the accomplishment of this paper. We would like to thank Lydia
Althea C. Brazil, RPh and Gruppo Medica for their assistance with the chemicals needed,
Engr. Antonio P. Rufino for his financial assistance and overall transportation, and the
University of the Philippines Los Baños, Department of Biotechnology in helping us with
their various services. We would also like to thank Ms. Phebe Mendoza, MSc, Ms. Leah C.
Madrazo, Doc. Kerry Cabral, and De La Salle University Integrated School, Chemistry
Department Research Laboratory, whom without their guidance, this research would not
have been possible.

Running Head: BIO-PIEZOELECTRIC NANO GENERATOR ii
Abstract
Bio-piezoelectric nanogenerators have been one of the greatest innovations in the
biomedical field due to their ability to produce and generate significant power through the
use of mechanical energy without inflicting damage to the environment. Highly efficient
piezoelectric materials in the current generation contribute various unavoidable problems to
the environment due to high toxicity and complexity in synthesis. In this study, the
researchers developed a potential biocompatible yet cost-efficient nanogenerator that could
provide sufficient electrical energy using extracted collagen from the fish bones of tilapia
(Oreochromis mossacimbus), called the Fish Bone Bio-piezoelectric Nanogenerator
(FBBPNG). The highest generated output voltage of the FBBPNG was ~6.5V through
bending movements, with a steady voltage of ~3.06V under mechanical stress of 5.32kPa.
When 0.76kPa was applied onto the FBBPNG, it was able to generate 0.560V, and at
4.56kPa, it was able to generate 2.57V. With this, it can be observed that the pressure
applied onto the device and the voltage output has a direct relationship, however, there is a
certain pressure threshold that the device can handle before the generated output voltage
begins decreasing. Consequently, FBBPNG is also sensitive on several body motion
activities such as heel pressing, foot pressing from the heel side, and wrist movement which
can evidently be a source of electricity. Due to its biocompatibility, the FBBPNG would be
of great interest in the medical field.
Keywords: Bio-Piezoelectricity, Fish Bones, Nano-Generator
,
Running Head: BIO-PIEZOELECTRIC NANO GENERATOR iii
Running Head: BIO-PIEZOELECTRIC NANO GENERATOR iv
Table of Contents
Acknowledgment i
Abstract ii
Table of Contents iii
Chapter 1 INTRODUCTION 1
1.1 Background of the Study 1
1.2 Problem Statement 5
1.3 Objectives 6
1.4 Significance of the Study 7
1.5 Scope and Delimitations 7
1.6 Definition of Terms 9
Chapter 2 LITERATURE REVIEW 13

2.1 Energy 13
2.2 Types of Energy 14
2.3 Electromechanical Conversion 16
2.4 Piezoelectricity 17
2.5 Piezoelectric Materials 18
2.6 Collagen Extraction 21
2.7 Demineralization or Desalination 22
2.8 Energy Harvesting System 24
2.9 Overall Synthesis 28
Chapter 3 METHODOLOGY 31
3.1 Materials/Equipment 31
3.2 Procedure 33
3.3 Safety Precautions 35
3.4 Research Methodology Flowchart 35
3.5 Equations for Data Collection 36
3.6 Research Design 37
3.7 Data Analysis Strategy 38
Chapter 4 RESULTS AND DISCUSSION 40

4.1 Energy Harvesting Performance and Working Mechanism 40
4.2 Human Locomotion and Human Activities 43
Chapter 5 CONCLUSION AND RECOMMENDATIONS 45
References 47
Running Head: BIO-PIEZOELECTRIC NANO GENERATOR v
LIST OF TABLES
4.2 Table 1 .................................................................................................................... 31

4.3 Table 2 .................................................................................................................... 32
4.4 Table 3 .................................................................................................................... 33
4.5 Table 4 ..................................................................................................................... 35
4.6 Table 5 ..................................................................................................................... 41
LIST OF FIGURES
4.7 Figure 1 ................................................................................................................... 34

4.8 Figure 2 ................................................................................................................... 36
4.9 Figure 3 ................................................................................................................... 40
4.10 Figure 4 .................................................................................................................... 41
4.11 Figure 5 .................................................................................................................... 42
4.12 Figure 6 ................................................................................................................... 44
Running Head: BIO-PIEZOELECTRIC NANO GENERATOR 1
Bio-Piezoelectric Nano generator: Harnessing Electrical Energy
from Fish Bones of Tilapia (Oreochromis mossambicus)
1.1. Background of the Study
With increasing amounts of technology comes increasing amounts of consumed
electricity. Given that the world is an ever-evolving place, many aspects of it may both
devolve or evolve. However, focusing on the aspects which humanity has taken advantage
of, such as electricity, we have learned how to utilize various objects in order for us to
advance. According to the Global Energy Statistical Yearbook (2017), electricity
consumption has increased dramatically over the span of 13 years, from 2004’s 15,037
Terawatts per hour (TWh) to 2016’s 21,191 TWh. Following this, it is known that Asia has
also observed the largest increase in electricity consumption, from 4,433 TWh to 9,053 TWh
in 2004 and 2016 respectively. With this knowledge, it can be determined that Asia uses
more and more electricity for each year that passes, therefore, electricity generation needs to
keep up with the demand of electricity consumption.
This consumption of electricity will continue to proportionally increase as the
demand of innovation persists. Consequently, there is a need for us to keep up with the
demands of electrical consumption through various methods of generating electricity. The
world is using large amounts of energy every day, and sources of energy are limited which is
feared to be gone completely if the world continues to utilize them, particularly the
nonrenewable ones called fossil fuels, such as coal, oil, and wood.
Prior to the start of the industrial age, which was around 200 to 300 years ago,
humanity has met the vast majority of their energy needs with various renewable energy
resources such as wood for heat, water mills for grinding crops, or the wind to propel sailing
vessels. Through the discovery of fossil fuels cut as oil, natural gas, and coal, we have found
new sources of energy that have been pivotal in the establishment of industrialized
economies. Modern society has been fundamentally shaped by the use of fossil fuels, and we
are still very much dependent on these fuels for our basic energy needs. Essentially, fossil
fuels are the remains of ancient organisms, such as old plants and bacteria. Following
changes in the Earth’s climate and geology, a portion of this organic matter was sequestered
below the Earth’s surface under multiple layers of sediment and rock and has been exposed
to immense heat and pressure given off by the Earth’s interior for millions of years. This
process created the fossil fuels that we use today, resulting from the originally decaying
biomass. As a result, natural oil, gas, and coal deposits can be found in specific geological
sediments throughout the world (Missouri Department of Natural Resources [DNR], 2017).
Given that these fossil fuels are the main sources of heat and electrical energy, all
these fuels contain materials including metal, sulfur, and nitrogen compounds. When we
burn gasoline in our cars, we are, in essence, burning ancient plants. The chemical process is
known as combustion, and burning fossil fuels is the equivalent of burning a piece of
firewood from a tree that existed several million years ago. During combustion, pollutants
such as fly ash, sulfur oxides, nitrogen oxides, and volatile organic compounds are emitted.
Gross emission of pollutants is tremendous all over the world, and these pollutants are
present in the atmosphere in such conditions that they can affect both man and the
environment (Chmielewski, 2005).
It is known that majority of the world’s energy needs are currently being met by the
use of fossil fuels. Fossil fuels have proven to be an abundant source of energy and have
been critical in the development of our modern world. However, the use of fossil fuel also
raises serious environmental issues such as acid rain. Acid rain has been shown to have a
significant impact on both aquatic and forest ecosystems. Mercury emissions and nitrogen
oxide contributes to various ozone pollution and serious health issues. The mining,
extraction, and transportation of fossil fuels can cause various types of environmental
damage ranging from habitat loss to pollution and the contamination of ecosystems. In
addition to the negative effects of fossil fuels to the environment, they are a limited resource;
the world will not only require energy in the future, but the energy needed in the future is
destined to grow. This is why we must find other sources of power, such as renewable
energy, as we will eventually run out of fossil fuels (DNR, 2017).
Renewable energy utilizes natural resources such as the sun, the wind, the plants, and
moving objects. By developing technology that can harvest these natural resources, these
can be turned into energy that would serve as electricity. With refinements in technology,
the feasibility and cost of solar and wind power systems have become affordable. Along
with these refinements in technology, market systems are rapidly evolving in favor of
renewable energy system. Major contributions are also obtained from the use of large
hydropower and solar energy, wind energy, modern bio-energy, geothermal energy, and
small hydropower energy sources are being increasingly tapped. Such a situation calls for an
implementation of aggressive long-term renewable energy programs for domestic and
commercial purposes.
Moreover, previous researchers have applied the concept of piezoelectricity in
energy harvesting systems. The piezoelectric plate is a device that holds within it
piezoelectric materials such as Quartz, Barium Titanate, and PZT, that produces a particular
kind of electricity--piezoelectricity through “squeezing” the piezoelectric material inside the

piezoelectric plate (Woodford, 2017).
Though it is considered that multiple piezoelectric materials address the problems
concerning our environment, they also produce and bring problems of their own. Lead has
been expelled from many commercial applications and materials, but this is one of the many
common components that is most commonly used in piezoelectric ceramics such as Lead
Zirconate Titanate (PZT). According to Saito et al. (2004), these materials contain almost 60
percent lead, owing its concerns to toxicity.
Bio-piezoelectric materials are discovered as a response to the adverse effects of
man-made piezoelectric materials. Bio-piezoelectricity utilizes materials that are rich in
cellulose, collagen fibrils, silk, peptide fibrils and bacteriophages. Among these biological
piezoelectric materials, the most abundant is collagen, which is present in animal tissues
such as skin, tendon, cartilage, and bones. In recent years, researchers have been studying on
collagen from aquatic animals compared with livestock sources because of the safety
brought by the collagen on aquatic animals. However, because of the very low populations
of several aquatic animals, studies from these sources cannot be supported. Because of this,
tilapia is a good candidate for farming as it is considered as the second most important farm-
raised fish (Liu & Huang, 2016). In the Philippines, tilapia is considered as the top species
for inland fish production (Philippine Statistics Authority, 2016). Although waste products
from tilapia processing have been used initially, large amounts of fish heads and skeletons
are disposed of every year.
Tilapia bone is a good source of collagen for piezoelectricity. Bones are rich in type I
collagen which can cause spontaneous polarization and piezoelectricity (Ghosh & Mandal,
2016; Karan et al., 2018). Collagen fibers can be extracted through the process of
demineralization or desalination.
One application of this concept is the Bio-Piezoelectric Nano generator (BPNG)
made with fish scales (FSC) that was developed by Ghosh and Mandal (2016). The
researchers were able to develop a high-performance BPNG with FSC as its main
component, as FSC is composed of self-assembled and ordered collagen nano-fibrils. As a
power source, it is able to generate an output voltage of 4 V, but when four BPNGs are
assembled, they are able to cumulatively generate an output of 14 V that is able to power
over 50 blue LEDs instantaneously. Proving itself as a sustainable green power source for
the next generation.
1.2. Problem Statement
In modern times, almost all of our electricity is generated from fossil fuels such as
coal, oil and, natural gas. However, these sources of nonrenewable energy diminish over
time and can bring several disadvantages to the environment. Therefore, we need a source of
renewable energy that would have little to no impact to the environment. With that said,
several studies show the conversion of mechanical energy into electricity. Piezoelectricity is
the electricity converted from the mechanical stress placed on piezoelectric materials.
Different studies have already tested the efficiency of this process and have used different
piezoelectric materials, particularly lead zirconate titanate (PZT) for energy harvesting
devices. However, these materials have a very complex synthesis method, and are
considered as brittle, toxic and non-biodegradable which resist them for real life
sensible/potential applications, especially in healthcare and biomedical point, despite of their
high output abilities. In response to this, the researchers considered the utilization of
biodegradable and biocompatible piezoelectric materials that can be used for energy
harvesting. Biological piezoelectric materials rich in collagen fibrils such as fish scales were
utilized in many researches, but the researchers are proposing another source of collagen to
be used in generating piezoelectricity; fish bones. However, because of the very low
populations of several aquatic animals, studies from these sources cannot be supported
which makes Tilapia a good candidate for this research as it is considered as the second most
important farm-raised fish around the world and even here in the Philippines. In addition to
that, large amounts of fish heads and skeletons are disposed of every year which makes this
research also helpful to waste disposal. In this research, the researchers have explored the
potentiality of an inexpensive bio-waste, Tilapia (Oreochromis mossambicus) bones, as an
efficient piezoelectric material.
1.3. Objectives
The study aims to generate a Biological Piezoelectric Nano generator which utilizes
fish bones from Tilapia (Oreochromis mossambicus) as a source of collagen which can
convert mechanical stress into piezoelectricity.
Specifically, this study endeavors the achieve the following:
a) To fabricate the Fish Bone Bio-Piezoelectric Nanogenerator (FBBPNG)
b) To determine the electrical output and the pressure-voltage relationship of the
FBBPNG.
c) To apply the FBBPNG for human locomotion and different activities.

1.4. Significance of the Study
This technology would benefit the environment, different fields of research, and
other researchers. Our world is in dire need of another source of energy as the non-
renewable resources continues to diminish. At the same time, these resources contribute to
the carbon footprints in our environment. As this can be applied to energy harvesting
devices, this research will be able to give another source of electricity, which are fish bones
and at the same time, provide environmental sustainability as this new source will not
contribute to the problems in our environment. In addition to that, as the demand for the
small power that can be produced by piezoelectric generators increases, the production of
electricity from vibrations is pertinent for energy harvesting. Although there have been
previous studies regarding piezoelectric generators, used materials in these researches cause
various implications in our environment such as toxicity, limiting their application to other
fields. In line with this, this research provides a biodegradable source of piezoelectricity and
may be utilized for other fields of research, particularly in the medical field, as this is a
biodegradable, biocompatible, and inexpensive source of electricity. Moreover, this research
would generate a great deal of interest, not only among fellow researchers, but also the
community market that uses different energy harvesting devices as this research introduces
another source of piezoelectricity.
1.5. Scope and Delimitations
The main focus of this research is to generate a biological piezoelectric material
which utilizes fish bones as a source of collagen which can convert mechanical stress into
piezoelectricity. This conversion will be done through the use of a Bio-Piezoelectric

Nanogenerator. This technology can only generate one kind of electricity, piezoelectricity.
For the fabrication of the FBBPNG, the study utilized 1 mm thick copper sheets and
laminated polypropylene (PP) films with approximately 125 µm thickness and copper wires
attached on both sides of the copper sheet. This project also focused on the measurement of
the voltage output of the FBBPNG and the devices’ application to different human
locomotion. The researchers utilized a recycled bio-piezoelectric material from the bones of
Tilapia (Oreochromis mossambicus). Fish bones contain hydroxyapatite and type 1 collagen
which are responsible for the piezoelectric reaction from the mechanical stress exerted by
the external force. Due to possible chemical changes, the fish bones that will be used in this
research will be from fishes that are uncooked.
This study did not include the production of other types of electricity, such as current
electricity, hydroelectricity, static electricity, and solar electricity. The study did not tackle
other piezoelectric materials, such as Quartz, Gallium Orthophosphate, Barium Titanate,
Lead Zircon Titanate, Berlinite, and Aluminum Nitride, since collagens are from biowaste
which are highly available and possesses longitudinal piezoelectric sensitivity, making them
useful in this study (Karan et al., 2018). Other sources of collagens such as fish scale, fish
muscle, gelatin, egg white, egg shell membrane, etc. were disregarded. Fish bones coming
from the leftovers from food were utilized in this study. Although the researchers had
studied different factors that may affect the output performance of the device, the
researchers did not conduct a structural analysis and a morphological study on the
Piezoelectric Nano generator for the study of the crystalline nature and existence of highly
ordered protein collagen fibrils inside it that gives its piezoelectric effect. This study did not
measure the generated current of the fish bones, the piezoelectric coefficient of the
fishbones, and the electric storage potential of the device because of the availability and
resources of the researchers.
1.6. Definition of Terms
Acidification/Acidify. the process of converting or turning something into acid.
Antiresorptive. the tendency of slowing or blocking the resorption of the bone.
Battery. any of a class of devices that convert chemical energy directly into
electrical energy.
Biomass. organic matter, especially plant matter, that can be converted to fuel and is
therefore regarded as a potential energy source.
Bio-piezoelectricity. piezoelectricity generated using organic substances.
Collagen. the main structural protein found in skin and other connective tissues.
Combustion. chemical combination attended by production of heat and light.
Current. the rate of flow of electricity.
Deformation (Material Science). any changes in the shape or size of an object due
to an applied force.
Dipole. a molecule in which a concentration of positive electric charge is separated
from a concentration of a negative charge.
Efficiency (Economics). a measure of the input a system required to achieve a
specified output. A system that uses few resources to achieve its goals is efficient, in contrast
to one that wastes much of its input.
Electricity. a fundamental form of energy observable in positive and negative forms
that occurs naturally or is produced, and that is expressed in terms of the movement and
interaction of electrons.
Electroneutrality. the neutrality of electricity, especially as applied to the principle
that pure ionic substances are electrically neutral.
Endogenous. not attributable to any external or environmental factor.
Energy (Physics). the capacity for doing work. It may exist in potential, kinetic,
thermal, electrical, chemical, nuclear, or other various forms.
Equilibrium. a phenomenon in which all the forces that act upon an object are
balanced.
Kinetic Energy. a type of energy that is transferred from one object to the other
which causes its motion.
Mechanical Stress/Stress. a physical quantity that expresses the internal forces that
neighboring particles of a continuous material exert on each other, while strain is the
measure of the deformation of the material.
Nanogenerator. a type of technology that converts mechanical/thermal energy as
produced by small-scale physical change into electricity.
Osteoclast. a large multinucleate bone cell that absorbs bone tissue during growth
and healing.
Piezoelectricity. the appearance of an electrical potential (a voltage, in other words)
across the sides of a crystal when you subject it to mechanical stress (by squeezing it).
Polar. being or having a molecule in which there is an uneven distribution of
electrons and thus a permanent dipole moment.
Polarization. property of certain electromagnetic radiations in which the direction
and magnitude of the vibrating electric field are related in a specified way.
Polypeptide Chains. a chain that consists of smaller sub-units or amino acids that
are linked together in order to form various proteins.
Potential Energy. stored energy that depends upon the relative position of various
parts of a system.
Protease. an enzyme that breaks down proteins and peptides.
Renewable Energy. any naturally occurring, theoretically inexhaustible source of
energy, as biomass, solar, wind, tidal, wave, mechanical, and hydroelectric power, that is not
derived from fossil or nuclear fuel.
Resorptive/Resorption. the destruction, disappearance, or dissolution of a tissue or
part by biochemical activity, as the loss of bone or of tooth dentin.
Sequestered. to trap (a chemical in the atmosphere or environment) and isolate it in
natural or artificial storage area.
Triboelectric Effect (Triboelectricity). a type of contact electrification on which
certain materials become electrically charged after they come into frictional contact with a
different material.
Uniaxial. Having only one direction along which double refraction of light does not
occur.
Vibration. mechanical phenomenon wherein there is a back and forth movement that
occurs about an equilibrium point. It is also referred to as shaking, trembling, and
brandishing.
Voltage. force of an electrical current that is measured in volts.

CHAPTER 2
REVIEW OF RELATED LITERATURE

s
2.
2.1. Energy
2.1.1. Description.
It is evident that in our world today, almost everything works because of energy.
Energy, in physics, is defined as the capacity for doing work (Energy, 2018a), but this
definition of energy was not applied until rather late in the development of the science of
mechanics (Energy Conversion, 2018). Although this definition is widely used, somehow, it
is incomplete and misleading since this implies that all energy produces work, but in reality,
not all. There is really no unique, absolute or universal concept of energy leaving it with no
simple definition (Sefton, 2004). However, we can say that energy is a fundamental entity
whose availability and flow is needed for all natural or artificial phenomena (Carnegie
Mellon University, 2001). Energy is also defined as power derived from the utilization of
physical or chemical resources, especially to provide light and heat or to work machines
(Energy, 2018b). Which means it can be used to power up machines in our current
technology.
2.1.2. Forms of energy.
Energy exists in many different forms and can change or be converted from one
form to another, and this process is called energy conversion. These different forms include
gravitational, kinetic, thermal, elastic, electrical, chemical, radiant, nuclear, and mass
energy. Although energy can be converted to these different forms, it is within the constraint
of the conservation law. This is a fundamental law that holds for all natural phenomena
which states that the total energy remains constant regardless of the changes and the
transformation going on in nature. Although the total amount of energy in an isolated
system remains unchanged, there may be a great difference in the quality of different forms
of energy (Energy Conversion, 2018).
Forms of energy include mechanical energy and electrical energy. Mechanical
energy is the energy associated with the motion and position of an object (Nuclear Power,
2018). It is defined as the sum of the kinetic and potential energy (Mechanical Energy,
2015) and can be separated into transient and stored energy. Transient mechanical energy is
commonly referred to as work, which is the energy in motion or energy being transferred
from one place to another. On the other hand, stored mechanical energy is the stored energy
or energy contained within a substance which can exists in one of two forms, kinetic and
potential. Potential energy is defined as the energy stored in object subjected to a
conservative force, while kinetic energy is defined as the energy stored in an object due to
its motion (Nuclear Power, 2018). Meanwhile, electrical energy is the energy of moving
electrons. The protons and electrons of an atom carry an electrical charge; protons having a
positive charge and electrons having a negative charge. Opposite charges attract each other
but the electrons on the outermost shell of the atom does not form a strong connection to the
protons which allows it to move from one atom to another, creating electricity (Demirel,
2012).
2.2. Types of energy.
The two main types of energy are primary and secondary. Primary energy is
captured directly from the environment while secondary energy is converted from the
primary energy to electricity or fuel. Primary energy can be classified into three different
groups, namely, non-renewable energy, renewable energy, and waste. Non-renewable
energy are those that diminish over time, not being able to replenish themselves. Examples
of these energy are fossil fuels, which include coal, nuclear, oil and, natural gas (Demirel,
2012). From the statistics of the International Energy Agency (2016), coal provided almost
majority of the energy used in our world, as well as coal and natural gas. Although it
benefits us as it is a valuable source of energy, the use of non-renewable energy has its
disadvantages regarding its effect on the environment and its sustainability. The use of fossil
fuels upsets the Earth’s balanced carbon budget, which contributes to temperature rising
faster than organisms can adapt (Morse, 2013). As a result, renewable energy was
discovered as an alternative source of energy. Renewable energy comes from sources that
naturally renew and regenerate. Examples of these energy are biomass, hydropower,
geothermal, wind and solar energy. The use of these kind of electricity reduces carbon
impact in the environment as these sources of energy do not directly emit greenhouse gases.
But as it is positive for our environment, it is also quite disadvantageous due to its high
price and its inefficiency (U.S. Energy Information Agency [EIA], 2017).
Secondary energy, on the other hand, are those energy created from primary sources
through energy conversion. Electricity, which we commonly use today to use devices like
televisions, computers, and cell phones which cannot be directly powered with other energy
sources, is considered as a secondary energy. Electricity is transformed from primary
sources such as coal, raw oil, fuel oil, natural gas, wind, sun, streaming water, nuclear
power, gasoline, and etc. Other examples of secondary energy are refined fuels such as
gasoline and synthetic fuels like hydrogen gas (EIA, 2017).

2.3. Electromechanical Conversion
2.3.1. Description and process.
The human, first of all, is a mechanical entity that lives in a mechanical environment.
From there, we can derive that man’s need for power of machines and industrial processes is
satisfied greatly by the conversion of mechanical energy to electric energy or vice versa.
Electromechanical conversion exists because of man’s basic need of mechanical assistance.
As long as it concerns the use of electrical sciences to serve basic human needs, it would fall
under the scope of electromechanical phenomena (Craig, 2002). When defined in
engineering, electro mechanics involves several processes and procedures drawn from
electrical and mechanical engineering. The process of electromechanical conversion
involves the interchange of energy between mechanical and an electrical system
(Electromechanical, 2018). This process is considered as reversible, as it can be transferred
back and forth between the electrical and mechanical system. However, as the conversion
occurs, some of the energy will be converted into heat and will be lost from the system
forever (Hiziroglu, 2010).
In a magnetic field, conductors are subjected to the action of electromagnetic forces
causing them to move. The motion of a conductor in a magnetic field causes induction of
electrical field and the electrical current existing in the conductor is a directed motion of
electrical charges. There are many ways to convert mechanical energy to electrical energy,
one of which is the application of pressure to a certain surface. This is called the
piezoelectric effect, or the induction of charges on the surface of a crystal by the application
of mechanical stress (Vukosavic, 2012). According to Ledoux (2011), this phenomenon is
generally referred to as the appearance of an electrical potential across the sides of a crystal
when you subject it to mechanical stress. By applying pressure on a surface of a crystal, it
causes variable forces to act on its surfaces, giving rise to voltage between surfaces,
producing piezoelectricity (Vukosavic, 2012).
2.4. Piezoelectricity
2.4.1. Description and process.
Piezoelectricity is the product of a phenomenon called piezoelectric effect.
Piezoelectricity can be generated by squeezing certain crystals, such as quartz, and make
electricity flow through them. The reverse is usually true as well: if you pass electricity
through the same crystals, they “squeeze themselves” by vibrating back and forth. These
reactions, electrical field and mechanical behavior, can be in either direction. The direction
of the electrical field may lead to a mechanical reaction depending on the material
(Woodford, 2017). In order to produce the piezoelectric effect, the polycrystalline is heated
under the application of a strong electric field. The heat allows the molecules to move freely
and the electric field forces all of the dipoles in the crystal to line up and face in nearly the
same direction.
In 1880, Pierre Curie and Jacques Curie discovered the piezoelectric effect through
the joint knowledge of what they knew about pyro electricity and about structures of crystals
to demonstrate the effect with tourmaline, quartz, topaz, cane sugar, and Rochelle salt. The
converse effect of producing electricity through the application of mechanical stress to the
crystals was discovered lately by Gabriel Lippmann in 1881 through the mathematical
aspect of the theory. The first applications were made during World War I with piezoelectric
ultrasonic transducers (Leodux, 2011).

2.4.2. Related studies of powering gadgets using piezoelectricity.
There are all kinds of situations where people need to convert mechanical energy
into electrical signals or vice-versa. In ultrasound equipment, a piezoelectric transducer
converts electrical energy into extremely rapid vibrations which can be used for scanning
and cleaning. In a microphone, sound energy is being converted into electrical energy by
piezoelectric crystals. This is through simply sticking the vibrating part of the microphone to
a crystal and, as pressure waves from one’s voice arrive, it will make the crystals move back
and forth, generating electrical signals. In a quartz clock or watch, the reverse-piezoelectric
effect is used to keep time precisely. Electrical energy from a battery is fed into a crystal to
make it oscillate thousands of times a second. Piezoelectricity is also used in spark lighters
for gas stoves and grills. When one presses the switch, what the individual is doing is
squeezing a piezoelectric crystal, generating voltage, and making a spark fly across a small
gap (Woodford, 2017).
2.5. Piezoelectric materials.
2.5.1. Description and examples.
There are all kinds of situations where people need to convert mechanical energy
into electrical signals or vice-versa. In ultrasound equipment, a piezoelectric transducer
converts electrical energy into extremely rapid vibrations which can be used for scanning
and cleaning. In a microphone, sound energy is being converted into electrical energy by
piezoelectric crystals. This is through simply sticking the vibrating part of the microphone to
a crystal and, as pressure waves from one’s voice arrive, it will make the crystals move back
and forth, generating electrical signals. In a quartz clock or watch, the reverse-piezoelectric
effect is used to keep time precisely. Electrical energy from a battery is fed into a crystal to
make it oscillate thousands of times a second. Piezoelectricity is also used in spark lighters
for gas stoves and grills. When one presses the switch, what the individual is doing is
squeezing a piezoelectric crystal, generating voltage, and making a spark fly across a small
gap (Woodford, 2017).
2.5.2. Effects of man-made piezoelectric materials.
Although piezoelectric materials are considered to addressing the problems
concerning our environment, they bring problems of their own. Lead has been expelled from
many commercial application and materials but this is one of the common components in
most commonly used piezoelectric ceramics, lead zirconate titanate. These materials contain
almost 60 weight per cent lead, owing its concerns to its toxicity (Saito et al., 2004).
Because of this, the possible applications of piezoelectric materials have been limited and
not mostly used for medical purposes, since even the existence of a minimum amount of
toxicity can be dangerous to patients which can cause poisoning. Which has an adverse
effect on intellectual and neurological development, and can even lead to death (Gordon,
Taylor, & Bennett, 2002). In addition to that, piezoelectric materials are fabricated or made
in a very precise and in a complex way (Ledoux, 2011). Because of this, its cost factor also
needs to be addressed.
2.5.3. Bio-piezoelectric materials.
2.5.3.1. Description.
With the word ‘bio’ which means life, bio-piezoelectric materials are composed of
parts of living organisms such as animals and plants. These materials can still generate
electricity due to the presence of electric dipoles. Because of the adverse effects of man-
made piezoelectric materials, such as lead zirconate titanate (PZT) and at the same time,
fully addressing the environment problems that we are facing today, bio-waste materials are
used as piezoelectric materials. The most easily available nature driven biodegradable and
biocompatible materials such as cellulose, collagen fibrils, silk, peptide fibrils and
bacteriophages are used as a green piezoelectric material. Due to its availability and its
sensitivity when exposed to mechanical stress, collagen-based materials are the ones mostly
used for various researchers in the present time (Karan et al., 2018).
2.5.3.2. Collagen.
Among these biological piezoelectric materials, the most abundant is collagen, which
is present in animal tissues such as skin, tendon, cartilage, and even in bones (Ghosh &
Mandal, 2016). There are many different types of collagen but the researchers will focus on
type I collagen which are commonly present in bones. Present in this type of collagen are
two similar alpha-1 (I) chains and one dissimilar alpha-2 (I) chain are present, wherein each
contains repeated triplet amino acid motif sequences of Gly (Glycine)-X-Y, where X and Y
are frequently proline (Pro) and hydroxyproline (Hyp), respectively. By twisting the alpha
chains together, a triple-helical structure will be formed. The quarter staggered arrangement
of collagen molecules gives collagen fibrils further assemble into a collagen fiber and gives
it an axial periodic structure (Bou-Gharios & Crombrugghe, 2008). This intrinsic presence
of polar uniaxial orientation of hydrogen bonding motifs between the polypeptide chains
acts as molecular dipoles which cause spontaneous polarization and piezoelectricity (Ghosh
& Mandal, 2016; Karan et al., 2018).
2.5.3.3. Fish bones.
The majority of bones consist of bone matrix which are both organic and inorganic in
nature. Forming the inorganic part of the bone matrix is the crystalline structure called
hydroxyapatite. On the other hand, the organic part of the bone matrix is called Type 1
collagen. Both of these components have been discovered to be responsible for
piezoelectricity in bones. When mechanical stress is applied, the collagen molecules which
consist of charge carriers move from the inside to the surface of the specimen. This
movement across the bone produces piezoelectricity. According to Muralidharan, Shakila,
Sukumar, and Jeyasekaran (2011), high collagen yield could be obtained from trash fish,
leather jacket (Odonus niger) through the process of extraction which uses acetic acid with
pepsin. The collagen that is most abundant from the leather jacket was Type I collagen
which is said to be one of the parts that is responsible for the generation of electricity. Since
the production of piezoelectricity using bones entails the need of Type 1 collagen, fish
bones, on the other hand, are also rich in Type 1 collagen which can then be used as a
piezoelectric material.
2.6. Collagen Extraction
2.6.1. Description.
Muralidharan et al. (2011) conducted a study focusing on the extraction of collagen
contents from skin, bones, and muscles from trash fish, leather jacket (Odonus niger).
Before the extraction, the skin, bones, and muscles of the fish were cut into small pieces then
treated with 0.8M sodium chloride (NaCl) at a ratio of 1:6 (w/v) for ten minutes in order to
remove the impurities, repeating the process for three times and then cleaned with cold
distilled water. After finishing the process, the fish should now be treated with 0.1M sodium
hydroxide at a ratio of 1:10 (w/v) for three days in order to remove the non-collagenous
proteins and to prevent the effect of endogenous proteases on the collagen contents,
changing the solution every day and then washing it with cold distilled water as a final step.
For the collagen extraction proper, the collagen was extracted using acetic acid following
three different methods. In the first method, 10 volumes of 0.5M acetic acid for extracting
acid soluble collagen (ASC) twice. In the second method, ASC was extracted first and an
addition of 0.1% (w/v) pepsin to 0.5M acetic acid was made to extract pepsin soluble
collagen contents (PSC). The purpose of adding pepsin was to split the helical region,
telopeptide. In the third method, PSC was extracted twice. After each extraction, the solution
obtained was centrifuged at 9,000 g for twenty minutes at 4 ° C. After that, the supernatant
was salted out with 2M NaCl for 24 hours at 4 ° C. Then, the precipitated collagen will be
centrifuge again at 9,000 g for 20 min at 4 °C. The residue was place into the dialysis
membrane-110 bags and then dialyzed against 0.02M phosphate buffer (pH 7.2) for 24 hours
at 4 °C. The dialyzed samples were then lyophilized and frozen at −20 °C for analysis. The
yield of collagen was calculated based on the hydroxyproline content in extracted collagen
to that of the raw material in which results from the yield of collagen acquired was varying
from 46.48% to 70.94% depending on the method used.
2.7. Demineralization or Desalination.
2.7.1. Description.
Demineralization is the process of putting away the minerals of the hard tissues of
living organisms. These hard tissues are composed of insoluble calcium salts of carbonate,
silicate, and phosphate ions. Thermodynamics and accurate measurements of the kinetics is
needed for drawing mechanistic conclusions concerning demineralization. The rate of

demineralization depends mainly on the temperature and concentration of the acid, but since
cellular definition and enzymes are adversely affected if the temperature is raised, or when
the concentration of the acid is increased, demineralization is usually carried out below or at
a room temperature and a concentration of mineral acid above 5 percent of commercial nitric
acid (55 percent) is seldom used. Under these conditions, even small specimens of bone
must be immersed in acid for days so that they will be completely demineralized. The
mechanism of demineralization is complex and efforts are needed towards obtaining a
thorough understanding of the process (Ehrlich et al., 2008). The process of demineralization
can actually be categorized into two main groups; natural and artificial. Natural
demineralization is the erosion of certain biominerals such as bacteria, fungi, algae,
foraminifera, sponges, polychaetes, and mollusks which results from the production of
mineral and organic acids, acidic polysaccharides, and enzymes. On the other hand,
examples of artificial demineralization include the demineralization of bone, dentin and
enamel, and skeletal formations of corals and crustacean (Ehrlich et al., 2009).
2.7.2. Bone Demineralization.
A bone is a highly structural composite material and its demineralization dated back
to 1889 upon exploration of using demineralized bone for surgical implantation (Ehrlich et
al., 2009). This process involves the acidification of the isolated extracellular
microenvironment, mediated by a vacuolar enzyme in the cell’s ruffled membrane known as
H+ -adenosine triphosphatase (H+ -ATPase). An energy-independent Cl/HCO3 2 exchange
on the cell’s antiresorptive surface maintains the intraosteoclastic pH in the face of abundant
proton transport. In the end, a ruffled membrane Cl channel, charge-coupled to the H+ -
ATPase, preserves its electroneutrality resulting in the secretion of HCl into the resorptive
microenvironment, generating a pH of approximately 4.5. This acidic environment prepares
the bone mineral to demineralize the organic component of bone which is subsequently
degraded by a lysosomal protease (Ehrlich et al., 2008).
In a study conducted by Liu and Huang (2016), they stated that desalination or
demineralization is an important process of bone collagen extraction. Their process of
desalination involves ethylenediaminetetraacetic (EDTA) acid solution and hydrochloric
acid in which they soaked the collagen from tilapia bones. Results showed that desalination
with HCl was fast, but collagen yield was low. In comparison, collagen yield in the EDTA
solution was higher than those from the HCl-treated fish bone, making it more effective in
bone collagen extraction. The same process was used in Ghosh and Mandal’s research about
the use of fish scales for a nanogenerator. The fish scales were also immersed in a solution
of EDTA to obtain transparent and flexible demineralized fish scales. After demineralizing
the fish scales, the collagen layers present in it serve as active piezoelectric element in the
resulting fish scale which made it transparent and roll able for a flexible Piezoelectric Nano
generator (Ghosh & Mandal, 2016).
2.8. Energy Harvesting System
2.8.1. Description.
Energy harvesting is the conversion of ambient energy present in the environment
into electrical energy. It basically has an identical concept when compared to large-scale
renewable energy generation, such as solar or wind power, but very different in scale. When
we say large-scale power generation, this typically refers to generating power for a city, a
town, an entire village. But when we refer to energy harvesting, we focus on little things,
from the large-scale megawatts of power, to the small-scale micro- to milli-watts
(Ka'zmierski & Beeby, 2010).
Energy harvesting development has been inspired by the rapid increase in numbers
of various autonomous wireless electronic systems. Examples of these systems are wireless
sensor nodes that combine together to form wireless sensor networks. By definition, the
system being talked about is wireless, and cannot be plugged into a main supply, power has
to be provided locally, but no direct wired connection.
2.8.2. Kinetic energy harvesting.
The human body is full of potential energy and when used, becomes kinetic or
mechanical energy. Mechanical energy can be found virtually anywhere, as all you need for
mechanical energy is something that moves, something that vibrates. Wireless sensor
networks may be installed on these sources of mechanical energy, which makes converting
mechanical energy from ambient vibration into electrical energy quite enticing. The
frequency of mechanical excitation depends on the source: less than 10 Hz for human
movements, and over 30 Hz for machinery vibrations, these devices are what are known as
kinetic energy harvesters, or vibration power generators (Zhu & Beeby, 2011, in Kazmierski
& Beeby, 2011).
2.8.3. Examples of energy harvesting systems using piezoelectric materials.
There are multiple energy harvesting systems which have one thing in common:
renewable energy. In order to overcome our power needs, we have to generate electricity in
every way possible, and one of these ways is through stepping on stairs. One design of an
energy harvesting system is a “Staircase Power Generation System” as invented and
designed by Ramesh and Mathew (2014). Whenever a person would step on the stairs, that
step would actually go down, and spring back up, but this spring would send the mechanical
energy to a flywheel through a connecting rod, then the crank shift would then help the
flywheel rotate the belt, thus causing the 12V DC motor to collect and harvest the
mechanical energy, now becoming piezoelectricity.
2.8.4. Examples of energy harvesting systems using bio-piezoelectric materials.
Multiple bio-piezoelectric energy harvesting has been developed, using different
kinds of bio-piezoelectric materials such as fish scales, eggshell membranes, and onion skin.
These materials are all rich in collagen. The highly stable crystalline structure of the triple
helix polypeptide chains of the collagen nano-fibrils, along with the amino acids that come
with it, is what allows for these materials to produce the piezoelectric effect (Ghosh &
Mandal, 2016).
2.8.4.1. Fish scales.
Ghosh and Mandal (2016) conducted a study wherein they developed an efficient
flexible high-performance Bio-Piezoelectric Nano generator (BPNG) with bio-waste
transparent fish scales (FSC) as their main bio-piezoelectric material. In this study, it is
demonstrated how the FSC that is composed of self-assembled and ordered collagen nano-
fibrils can serve as a self-poled piezoelectric component, which exhibits an intrinsic
piezoelectric strength of -5.0 pC/N. Just like regular piezoelectric energy harvesting systems,
the BPNG is able to sense several types of ambient mechanical energies such as human
locomotion, machine and sound vibrations, as well as the flow of the wind.
The FSCs were collected from the local fish processing market and washed
thoroughly with deionized water followed by a solvent system of NaOH, 1.0 M NaCl, 0.05
M Tris HCl, and 20.0mM EDTA. The FSCs were then immersed in a solution of 0.5M
EDTA to obtain transparent and flexible demineralized FSCs. This causes the collagen
layers to serve as an active piezoelectric element in the FSC which is both transparent and
roll able, allowing for a BPNG to be synthesized. To develop the said BPNG, gold
electrodes of 90nm thickness have been made on both sides of FSC by the process of
sputtering, followed by lamination with the PP film with a thickness of around 125 µm.
Under a constant compressive normal stress of 0.17 Mpa, the BPNG is able to generate an
output voltage of 4V, a circuit current of 1.5µA, and a maximum output power density of
1.14µW/cm2. Integrating four BPNGs produces an output voltage of 14V, powering up to
more than 50 blue light emitting diodes instantaneously, thus proving to be an effective
sustainable green poawer source.
2.8.4.2. Eggshell membranes.
As bio-piezoelectric materials have been considered as excellent energy harvesting
sources due to their non-toxic and biocompatibility nature, Karan et al. (2018) have
developed an eggshell membrane Bio-Piezoelectric Nano generator (ESMBPNG). In this
study, the researchers have explored the potentiality of an inexpensive and bio-waste porous
eggshell membrane (ESM) as an efficient piezoelectric material which has a piezoelectric
strength of 23.7pC/N.
Fresh chicken eggs were broken and emptied, the eggshells were then washed with
deionized water, and the shell membrane was peeled out from the eggshell. The ESM was
dried at 60°C in an air-oven for an hour. Both sides of the ESM were attached with gold
fabrics serving as conductive wiring, followed by the ESM being covered by a copper tape
serving as top and bottom electrodes, respectively. PP tape was used to cover it, relieving it
from any triboelectric that may be caused from single electrodes. The ESMBPNG was then
encapsulated by polydimethylsiloxane (PDMS: Sylgard, 184 Silicone Elastomer) and a
curing agent (10:1 wt/wt), and is then dried at 60°C for 30 minutes. The PDMS layer will
protect the device from any external environmental hazards such as mechanical stress,
humidity, and temperature during experiments. The ESMBPNG alone is able to produce
26.4V, with a current of 1.45µA and a maximum instantaneous power density of around
238.17µW/cm3. When five ESMBPNGs are assembled, it provides an output voltage of
approximately 131V, lighting up more than 90 green LEDs.
2.8.4.3. Onion skin.
In this study, Maiti et al. (2017) tackled the development of non-toxic, ultrasensitive,
and flexible bio-inspired piezoelectric nanogenerators. As high-performance
organic/inorganic materials based piezoelectric nanogenerators suffer from several
unavoidable problems such as complex synthesis and high toxicity, a bio-piezoelectric
nanogenerator (BPNG) using naturally abundant cellulose fibrous untreated onion skin (OS)
has been developed. The OSBPNG generated an output voltage of 18 V, and a current of
166 µA, through a biomechanical force acting vertically on the OSBPNG. Amassing 6
OSBPNGs will grant an output voltage of 106 V and grant a 900 µA current, spontaneously
powering 73 LEDs that consisted of 30 greens, 25 blues, and 18 reds.
2.9. Overall Synthesis
The use of non-renewable resources such as fossil fuels as a source of energy has
been a part of our everyday living but are considered as not sustainable (EIA, 2018; Demirel,
2012). In addition, it contributes to the carbon footprints in our environment (Morse, 2013).
As a result, the use of renewable energy as a source of energy has been discovered as an
alternative to the non-renewable resources (EIA, 2017). Renewable energy consists of the
process of converting one form of energy to another, or what we call energy conversion.
Energy exists in many different forms and examples of these are mechanical and electrical
energy (Energy Conversion, 2018). These two forms of energy are commonly used in
renewable energy in a process called electromechanical conversion or the process of
converting mechanical energy into electrical energy (Electromechanical, 2018). There are
many ways of electromagnetic conversion and one of these processes is the application of
pressure in a surface, also called as the piezoelectric effect. The electricity generated from
this process, also called as the piezoelectricity, can be generated by squeezing certain
crystals, commonly known as piezoelectric materials (Vukosavic, 2012; Ledoux, 2011).
Different piezoelectric materials exist in our world today and can occur either man-made or
natural. Man-made piezoelectric materials, such as lead-zirconate titanate, are mostly used in
researches particularly in different energy harvesting system (Ledoux, 2011). However,
these materials have a very complex synthesis method, and are considered as brittle, toxic
and non-biodegradable which resist them for real life sensible/potential applications,
especially healthcare and biomedical point, despite of their high output abilities (Ledoux,
2011; Saito et al., 2004; Gordon, Taylor, & Bennett, 2002). In response to this, bio-
piezoelectric materials were used to counter the adverse effects of man-made piezoelectric
materials. These materials are rich in biodegradable materials and an example would be
collagen fibers, which are considered as the most abundant piezoelectric biomaterial present
mostly in animal tissues such as skin, tendons and especially, bones (Karan et al., 2018).
Bones consist of bone matrix which are both organic and inorganic, and the organic part of
the bone matrix is called Type 1 collagen (Wang, 2013). Collagen can be extracted from
bones through the process of demineralization or desalination. Present in collagen are polar
uniaxial orientation of hydrogen bonds between polypeptide chains which acts as molecular
dipoles resulting to polarization and piezoelectricity (Bou-Gharios & Crombrugghe, 2008;
Ghosh & Mandal, 2016; Karan et al., 2018). Because of this, Bio-Piezoelectric Nano
generator are now utilized in energy harvesting (Ghosh & Mandal, 2016; Maiti et al., 2017;
Karan et al., 2018). In line to this, the researchers are proposing the utilization of another
source of collagen, fish bones, in Bio-Piezoelectric Nanogenerator.

CHAPTER 3
METHODOLOGY
3.
3.1. Materials/Equipment
A list of materials and equipment that were used in the research is presented by the
following tables and is classified based on the four stages of our research procedure. Table 1
shows the materials, chemicals, and equipment that were used for the preparation and
extraction of the collagen from the tilapia bones. Table 2. shows the materials that were used
during the fabrication of the FBBPNG. Table 3. shows the materials and equipment that
were used for the data collection stage from the FBBPNG device.
Table 1
List of Materials, Chemicals and Equipment to be Used During the Preparation and
Extraction of Collagen from Tilapia (Oreochromis mossambicus) Bones
Materials, Chemicals, and Function/Use

Equipment
Materials
40-mesh screen Used for the filtration of crushed bones
whose diameters would be 0.0164 inches
and above
Mortar and Pestle Used to deteriorate the bone structure
while still keeping the collagen amount
intact
Chemicals
Sodium Hydroxide (NaOH) Used for cleaning the bones and remove
the non-collagenous proteins present.
Ethylenediaminetetraacetic acid Essential for the desalination or the
(EDTA) process in which excess sodium chloride,
minerals, and impurities would be

removed.
Acetic Acid (CH3COOH) Used for extracting and solubilizing the

collagen from the dry, desalinated bone
powder
Pepsin Used to break down the proteins into
smaller peptides, allowing the collagen to
be more prevalent.
Equipment
Freeze Dryer (Virtis Freezemobile Used to freeze-dry the fishbones,
25SL) removing excess water from the dry
bones without excessive heating, while
enhancing its stability for preservation.
Centrifuge (Hettich EBA-20) Used to separate liquids from solids and
even liquids at different densities by
using centrifugal force made from
spinning a vessel at high speed.
Table 2
Materials to be Used During the FBBPNG
Materials Function/Use
Copper Wire Functions as conducting wires for the
bio-piezoelectric material
Copper Tape Functions for conduction and attachment
of wires to the bio-piezoelectric material
Polypropylene (PP) Film (125 µm) Functions for protection from any
triboelectric effect caused by different
environmental hazards
Table 3
Materials and Equipment to be Used for the Data Collection from the FBBPNG
Equipment Function/Use
Oscilloscope (Agilent InfiniiVision Used to measure the electrical output of
DSO-X 2002A) the device; used to measure the charging
voltage of the device
Multimeter Used to measure the charging voltage of
the device
3.2. Procedure
The experimental procedure was categorized into three parts; the preparation and
extraction of collagen from of tilapia (Oreochromis mossambicus) bones, the fabrication of
the FBBPNG, and the collection of data from the FBBPNG.
3.2.1. Preparation and extraction of collagen from the tilapia.
The tilapia skeletons were collected which contained residual bones and meat from a
fish market and stored at-18 °C in a freezer for 1-3 d to preserve its freshness. It was then
thawed at room temperature (23 °C) before scraping residual fins and small portions of skin
from the bones using the scalpel. Afterwards, the bones were cleaned from different non-
collagenous proteins using 0.1 M NaOH, and water, filtered, washed with water, dried at
room temperature, crushed to obtain a small residue with a crusher, and then filtered using
40-mesh screen. In preparation for the collagen extraction, the residue was softened by
decalcification, using 0.5 M EDTA for 9 days, changing the EDTA solution per day. The
decalcified bones were washed with distilled water at 4 °C and then lyophilized for 1-2 d,
removing water from the dry bones without excessive heating, enhancing its stability for
preservation. The dry bones were extracted with 200 mL of 0.5 M acetic acid, having 20 mL
acetic acid per gram of bones, for 24 h. To separate the solid from the liquid substance, the
solution was centrifuged at 5,000 rpm for 20 m at 4°C. The residues were then re-extracted
and mixed again with the same amount and concentration of acetic acid, and centrifuged at
the same conditions. After that, the residue was re-dissolved in 0.5 M acetic acid, salted out
and centrifugated thrice and subsequently re-dissolved in 0.5 M acetic acid again with
1000U pepsin at 4 °C for 96 h. Again, the viscous solution was centrifuged at 10,000 rpm
for 20 m at 4 °C, separating the collagen from the aqueous solution. The collagen was
dialyzed against distilled water and then lyophilized. After demineralization, the collagen
layers collected served as an active piezoelectric element that makes it suitable for a
FBBPNG.
3.2.2. Fabrication of the fish bones bio-piezoelectric nanogenerator.
Copper sheets were attached on both sides of the Fish bone collagen (FBC), with a
1”x1” dimension followed by attaching copper wires (serving as a substitute for gold
electrodes) on both sides of the copper sheets. The device was covered using PP film. Figure
1 shows the design of the FBBPNG.
Figure 1. Fabrication of the Fish Bone Bio-Piezoelectric Nanogenerator (FBBPNG)

3.2.3. Collection of the data from the fish bones bio-piezoelectric nanogenerator.
For the collection of data from the FBBPNG, the device was tested using a
multimeter (U1230 Series, Synergy Technologies) and an oscilloscope (DSO-X 2002A,
InfiniiVision) for measuring its output performance and working mechanism from human
locomotion and activities, which is guided by the methodology of Maiti et al. (2017) and
Karan et al. (2018).
3.3. Safety Precautions.
In conducting this research, different safety precautions were followed especially
when handling the chemicals. Table 4. shows the list of chemicals that were used in the
experiments, their safety precautions and recommended personal protective equipment.
Table 4
List of Chemicals that were used, Their Safety Precautions and Recommended Personal
Protective Equipment (PPE)
CHEMICALS SAFETY PRECAUTIONS PERSONAL PROTECTIVE

EQUIPMENT (PPE)
Sodium Keep container dry. Avoid Gloves, Synthetic apron, Approved
Hydroxide contact with eyes and skin. and certified vapor and dust
(NaOH) Never add water to it. respirator, Splash goggles
Acetic Acid Keep away from heat. Do not Gloves (impervious), Synthetic
(CH3COOH) ingest. Avoid contact with eyes apron, Approved and certified
and skin. Keep in a cool area. vapor respirator, Splash goggles
Water (H2O) Quickly mop up if spilled. N/A
Absorb it with inert material.
Ethylenediamine Keep away from heat. Do not Gloves, Lab coat, Approved and
tetraacetic Acid ingest. Keep container tightly certified dust respirator, Safety
(EDTA) closed. Keep in cool area. glasses
Pepsin Avoid eye and skin contact. Do Gloves, Lab coat, Approved and
not breathe. Beware of the certified dust respirator, Safety
possible allergies involved. glasses
3.4. Research Methodology Flowchart
To have a graphical representation of the experimental procedure of this research, a
flowchart was created. Figure 2. shows the flowchart of the research procedure for the
FBBPNG.
Figure 2. Flowchart of the Research Procedure for the Fish Bone Bio-Piezoelectric
Nanogenerator (FBBPNG)
3.5. Equations for data collection.
3.5.1. Force from Mass and Acceleration.
Newton's second law of motion pertains to the behavior of objects for which all
existing forces are not balanced. The second law states that force is dependent on the mass
and the acceleration of an object, given by the equation, F = ma. Force is measured in
Newtons (N), mass in kilograms (kg), and acceleration in meters per second squared (m/s2).
This equation was used to find the force applied on the FBBPNG to be used in solving the
pressure applied on it.
3.5.2. Pressure from Force and Area.
Pressure and force are related, and so you can calculate one if you know the other by
𝐹
using the physics equation, 𝑃 = 𝐴. Because pressure is force divided by area, its meter-
kilogram-second (MKS) units are newtons per square meter, or N/m2. This equation was
used to identify the pressure-voltage relationship of the FBBPNG.
3.6. Research Design
The purpose of this study is to generate a Biological Piezoelectric Nano generator
which utilizes fish bones from Tilapia as a source of collagen which can convert mechanical
stress into piezoelectricity. To be able to construct a biological piezoelectric nanogenerator
which uses fish bones as the source of collagen, an experimental research design was
utilized. This research design enabled the researchers to design and build a nanogenerator
using a bio-waste piezoelectric material. An experimental design also aids in the
determination of the efficiency of the designed FBBPNG. To be able to gather data,
observations and calculations are both employed.

The participants of the study will be the collected bones of tilapia. Of all the
vertebrate fishes available in the market, the researchers used the Tilapia due to its
abundance and availability; thus, researchers used convenience sampling. The different
variables that were tested in this research were the energy harvesting performance of the
device through its output voltage, the energy harvesting performance of the device without
the PP tape, the output performance of the device under constant pressure using metal
weights, and its output performance under different human locomotion. The independent
variable in this study that will be varied by the researchers is the fish that will be used as the
source of collagen, Tilapia.
The aforementioned instruments that were used for testing the different variables are
only valid for a certain degree. Although there are better instruments that can be used to
gather data, these instruments were chosen by their availability and ease in operating.
3.7. Data Analysis Strategy
The data were analyzed based on the objectives of the research.
3.7.1. Polarity Reversion Test.
The voltage output of the device was measured with the reversion of the connection
of the wires. Different signs in the output voltage between the two trials would determine the
self-polarization of the FBBPNG. The data collected was presented using the time-voltage
graph displayed on the oscilloscope.
3.7.2. Electrical output performance with and without PP tape.
Initially, the voltage output of the fabricated FBBPNG with and without the PP tape
was measured and analyzed. An increase in output voltage without the PP tape would
determine that there has been an occurrence of triboelectric effect. The data collected was
presented using the time-voltage graph displayed on the oscilloscope.
3.7.3. Pressure-Voltage Relationship.
The FBBPNG was exposed to different amounts of pressure to measure the amount
of generated output voltage per certain amount pressure exerted. An increase or decrease in
output voltage as the applied pressure is increase would determine the pressure-voltage
relationship of the FBBPNG. The data collected was presented in tabular format and using a
line graph to show the relationship between the amount of electricity generated and amount
of pressure exerted for easier analyzation.
3.7.4. Harvesting performance in different human locomotion.
The feasibility of the fabricated FBBPNG to harvest the biomechanical energy for
developing human-based self-powered devices was clarified. This was shown through
exposing the device under different human locomotion such as heel pressing, foot pressing,
heel side pressing, below the shoe, wrist movement, neck movement, coughing, and
shouting. The data collected was presented using the time-voltage graph displayed on the
oscilloscope.
CHAPTER 4
RESULTS AND DISCUSSION

4.
4.1. Energy Harvesting Performance and Working Mechanism
In determining the energy harvesting performance and working mechanism of the
fabricated FBBPNG, the output voltage generated from simple motions applied on the
device, such as stretching and bending, was considered. Also, the relationship between the
applied pressure on the FBBPNG and its output voltage was studied.
4.1.1. Polarity Reversion Test.
Fig. 3. Output voltage from the polarity reversion test
Figure 3 shows the output voltage measured doing the connection polarity reversion
test. Two output voltages was measured, where the second output voltage was measured
with the reverse of the wires of the first one. Doing so, the output voltage generated form the
reversed polarity was negative, showing the dipole reversibility of the device. This proves
that the output voltage measured from the FBBPNG was from the FBC and not from any
electrostatic interactions. Although this was proven to be true, the self-polarization
mechanism in piezoelectric materials are yet to be established for their complex nature
(Saravanakumar, Soyoon, and Kim, 2014).

4.1.2. Electrical output performance with and without PP tape.
Fig. 4. Output voltage generated from bending the FBBPNG
Furthermore, we have checked the output performance of the device without the PP
tape, generating an output voltage of 7.08 V as shown in Fig. 4. There is an observed slight
increase of voltage than that of the highest output performance of the PP integrated device
(6.5 V) when removing the PP tape, leading us to assume that the use of PP tape could
minimize any triboelectric effect occurring in the device due to single electrode type of
nature.
4.1.3. Pressure-Voltage Relationship
5. Table 5
6. Crystal Deformation Per Applied Weight and Pressure
Mass of Object (g) Applied Pressure (kPa) Voltage (V)
50 0.76 0.560
100 1.52 1.13
150 2.28 1.53
200 3.04 1.69

250 3.80 1.77
300 4.56 2.57
350 5.32 3.06
Fig. 5. Pressure-Voltage Graph and Relationship
Table 5 shows the masses of the metal object applied on the FBBPNG, the calculated
pressure from these masses, and the output voltage of the device upon the application of mass.
Figure 1 shows the relationship between the applied pressure on the FBBPNG and its output
voltage upon application. As shown in Figure 1, the applied pressure of the FBBPNG is directly
proportional to the output voltage generated by the device, meaning that as the applied pressure
increases, the output voltage also increases, and if the applied pressure decreases, the output
voltage also decreases. This proves that the crystal deformation of the piezoelectric material is
dependent on the applied pressure on the device, which also changes its output performance
(Ghosh & Mandal, 2016; Maiti et al., 2017; Karan et al., 2018).
6.1. Human locomotion and human activities
We clarify the feasibility of our fabricated FBBPNG to harvest the biomechanical
energy for developing human based self-powered devices. Several body locomotion (heel
pressing, foot pressing from heel side, below the shoe, wrist movement, and neck movement)
and different human activities (coughing and shouting) have minutely been studied for
biomechanical applications.
The fabricated FBBPNG generated an output voltage of 0.8 V during heel pressing
while generating an output voltage of 2.13 V during foot pressing as shown in Figure 6a and 6b.
The higher output voltage generated under foot pressing than heel pressing is due to a higher
applied pressure or more crystal deformation of the former case. FBBPNG is also able to
harvest energy by wrist movement, as shown in Figure 6c, generating 5.11 V. Further, it is
highly sensitive to bending and stretching as presented in the previous discussion shown in
Figure 6d and 6e. The sensitivity of the FBBPNG was also checked by single finger pressing
and hand punching as shown in Figure 6f and 6g, generating 1.21 V and 3.50 V, respectively.
Consequently, the device displayed an output performance of 2.57 V from neck movement
(Figure 6h.), 0.16 V from coughing (Figure 6i.), and 200 mV by shouting “Stop!” and “Go!”
(Figure 6j.). Despite its low overall output performance, it showed a higher sensibility to human
locomotion and activities compared with the other bio-piezoelectric devices. All these findings
clearly demonstrate that FBPPNG can harvest energy during real life motions and could be
extended to various fields.

Fig. 6. The changes in output voltage versus time for monitoring motions in: (a) heel
pressing, (b) foot pressing front side below, (c) wrist movement (up and down), (d) bending
by fingers, (e) stretching, (f) finger pressing, (g) hand punching, (h) neck movement, (i) neck
movement, (j) and shouting “Stop!” and “Go!”, respectively. The inset of Figure 6a-c, and
g show the real photo images of the OSBPNG attached with respective specified portion.
CHAPTER V
Conclusion
In conclusion, fish bones, a naturally abundant biodegradable material, has been
shown to be a possible bio-piezoelectric material for the development of efficient energy
harvesting nanogenerators. The design of the FBBPNG is proven to be able to generate a
high amount of electricity for it to be used in energy harvesting. FBBPNG requires no
complex process for fabrication and is best fabricated covered in PP tape to prevent any
other sources of electricity other than the bio-piezoelectric material. In terms of voltage
output, it produced a lower voltage when compared to the previous bio-piezoelectric
devices. The highest voltage output generate by the FBBPNG by simple applied motions is
6.5 V from bending the FBBPNG. Despite its low output performance (~6.5 V), it can still
be used in small scale energy harvesting and commercialization. In theory, multiple
FBBPNG devices may be connected in series to be able to generate a higher output
performance based from the previous studies on bio-piezoelectricity. Also, it was found that
there is a direct relationship between applied pressure and the output voltage of the
FBBPNG, which was observed by applying different masses of metal weights on the device,
ranging from 50 up to 300 grams, as you can see on the screen. Lastly, FBBPNG showed a
high sensitivity on human locomotions. The fabricated FBBPNG generated a voltage output
of 0.8 V during heel pressing, 2.13 V during foot pressing, 5.11 V by wrist movement and
3.50 V by hand punching. Consequently, the device displayed an output performance of 2.57
V from neck movement, 0.16 V from coughing, and 200 mV by shouting “Stop!” and “Go!”.
From this, FBBPNG shows its capability to be used for realistic applications particularly
fields involving human motions.

Recommendations
The research would be recommended to be used and applied in energy-harvesting as
it proves to be able to generate electricity from the application of pressure on the FBBPNG.
This device could be extended to various fields, like inside the soles of the shoes, wheels
from the car, and different machines for the conversion of mechanical energy into electrical
energy to power up LEDs, LCD screen, and if possible, for mobile charging. At the same
time, being biocompatible in nature, it may still be used in the medical field apart from its
broad range application of the usual piezoelectric materials. But despite these possible uses
and applications and added knowledge on bio-piezoelectricity, this research is still subjected
to future studies to widen the understanding and if possible, increasing its output
performance. Firstly, a structural investigation and morphological study on the bio-
piezoelectric material can be done to further explain how the process of conversion from
mechanical energy to electrical energy happens in the fish bones. Second, the piezoelectric
coefficient of the device may be measured to add to the accuracy of the data collected as it
would be a good basis for comparison on the generated output voltage of the FBBPNG.
Third, other factors may be measured that can affect or is being affected by the output
voltage of the FBBPNG such as the dimensions of the bio-piezoelectric material, the
durability of the device, and even the materials used for the fabrication of the device. Fourth,
the generated current on the device must be established so that the FBBPNG may be used in
different circuits and other applications. Lastly, the charging capability and energy storage
of the device can be measured to extend the application of the FBBPNG in energy storage
and batteries.
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Senior High School Department

Bio-Piezoelectric Nano generator: Harnessing Electrical Energy

From Fish Bones of Tilapia (Oreochromis mossambicus)

Baliwag, Jesse Lambert R.

Brazil, Gian Miguel C.

Perez, Gabriel Francis D.

Rufino, Arvin Gabriel D.

Sambrano, Alphonse Rossaint V.

3rd Trimester, A.Y. 2018-2019

April 10, 2019

University of the Philippines Los Baños, Department of Biotechnology in helping us with

have been possible.

Bio-piezoelectric nanogenerators have been one of the greatest innovations in the

piezoelectric materials in the current generation contribute various unavoidable problems to

researchers developed a potential biocompatible yet cost-efficient nanogenerator that could

(Oreochromis mossacimbus), called the Fish Bone Bio-piezoelectric Nanogenerator

begins decreasing. Consequently, FBBPNG is also sensitive on several body motion

of great interest in the medical field.

Keywords: Bio-Piezoelectricity, Fish Bones, Nano-Generator

Table of Contents iii

Chapter 2 LITERATURE REVIEW 13

Chapter 4 RESULTS AND DISCUSSION 40

Chapter 5 CONCLUSION AND RECOMMENDATIONS 45

4.2 Table 1 .................................................................................................................... 31

4.7 Figure 1 ................................................................................................................... 34

Bio-Piezoelectric Nano generator: Harnessing Electrical Energy

from Fish Bones of Tilapia (Oreochromis mossambicus)

1.1. Background of the Study

With increasing amounts of technology comes increasing amounts of consumed

advance. According to the Global Energy Statistical Yearbook (2017), electricity

keep up with the demand of electricity consumption.

This consumption of electricity will continue to proportionally increase as the

demands of electrical consumption through various methods of generating electricity. The

environment (Chmielewski, 2005).

energy, as we will eventually run out of fossil fuels (DNR, 2017).

implementation of aggressive long-term renewable energy programs for domestic and

Moreover, previous researchers have applied the concept of piezoelectricity in

kind of electricity--piezoelectricity through “squeezing” the piezoelectric material inside the

piezoelectric plate (Woodford, 2017).

Though it is considered that multiple piezoelectric materials address the problems

percent lead, owing its concerns to toxicity.

Bio-piezoelectric materials are discovered as a response to the adverse effects of

man-made piezoelectric materials. Bio-piezoelectricity utilizes materials that are rich in

are disposed of every year.

One application of this concept is the Bio-Piezoelectric Nano generator (BPNG)

component, as FSC is composed of self-assembled and ordered collagen nano-fibrils. As a

the next generation.

1.2. Problem Statement

sensible/potential applications, especially in healthcare and biomedical point, despite of their

potentiality of an inexpensive bio-waste, Tilapia (Oreochromis mossambicus) bones, as an

efficient piezoelectric material.

convert mechanical stress into piezoelectricity.

Specifically, this study endeavors the achieve the following:

a) To fabricate the Fish Bone Bio-Piezoelectric Nanogenerator (FBBPNG)

b) To determine the electrical output and the pressure-voltage relationship of the

c) To apply the FBBPNG for human locomotion and different activities.

1.4. Significance of the Study