PASIG CITY SCIENCE HIGH SCHOOL

Rainforest Park, Maybunga, Pasig City, Philippines 1600

Telefax # 628-2177

Chitosan-based Siomai Bioplastic Plates Extracted

from Periplaneta americana

In partial fulfillment of the requirements in Practical Research II

Kenneth L. Maque

Mr. Avel Alcalde

11-Patella
 -

CHAPTER I

Background of the Study

Plastic is any synthetic or semisynthetic organic polymer. In other words,

while other elements might be present, plastics always include carbon and hydrogen.

While it may be made from just about any organic polymer, most industrial plastic is

made from petrochemicals (Helmenstine, 2019) that play a crucial role in various

manufacturing sectors however, potential environmental hazards associated with this

substance have raised concerns for societies. The industry releases large deleterious

substances as effluents into the atmosphere and generates solid waste such as plastic that

is difficult both to treat and to dispose of (Sharma et al., 2017). 8.3 Billion metric tons of

plastic has been produced since the time it was introduced in the 1950’s (Gooldjar, 2018)

and virtually every piece of plastic that was ever made still exists in some shape and form

until this day (Gonzaga, 2017). Plastic waste is one of many types of wastes that take too

long to decompose. Normally, plastic items can take up to 1,000 years to decompose in

landfills and even plastic bags that we use in our everyday life take anywhere from 10 to
1,000 years to decompose, and plastic bottles can take 450 years or more (Leblanc, 2019)

however, it spells big problem for human health. Ubiquitous marine plastic, for instance,

degrades and fragments into microplastics that can seep into the food chain and end up in

our bodies (Rosa-Aquino, 2019). According to National Geographic writer Laura Parker

(2018), “The miracle material that is plastic has made life possible. But more than 40% of

it is used just once, and it’s choking our waterways”.

Based on the study of Engler (2018), one of the ways to reduce plastic

pollution is plastic recycling, refers to the process of recovering waste or scrap plastic

and reprocessing the materials into functional and useful products (Leblanc, 2019)

however, encouraging individuals to recycle more will never solve the problem of a

massive production of single-use plastic that should have been avoided in the first place

(Wilkins, 2018). Many scientists have seen the potential in making plastic from

biodegradable objects and in 1926; Maurice Lemoigne invented the first bioplastic,

polyhydroxybutyrate that was made from the bacterium Bacillus megaterium (Barrett,

2018). Bioplastics simply refers to plastic made from plant or other biological material

instead of petroleum. It is also often called bio-based plastic. It can be either be made by

extracting sugar from plants like corn and sugarcane to convert into polylactic acids

(PLAs) that are commonly used in food packaging, or it can be made from

polyhydroxyalkanoates (PHAs) that are made from microorganisms that are used for

medical purposes (Gibbens, 2018). Recently, scientists at Harvard University’s Wyss

Institute for Biologically Inspired Engineering have developed a new biopolymer that

mimics the toughness, durability, and flexibility of natural insect cuticle. A biological
composite with a structure akin to plywood, insect cuticle consists of layers of the

polysaccharide polymer chitin and protein. The Wyss researchers extracted chitosan

sugar from discarded shrimp shells and engineered a thin, transparent film with a strength

equivalent to aluminium at only half the weight. The new Chitosan Bioplastic is

biocompatible, biodegradable, easily moulded into complex shapes, and inexpensive

because of the widespread availability of shrimp waste. Because of these beneficial

characteristics, the material is a potential replacement for conventional petroleum-based

plastic in applications ranging from consumer products to medical uses. Furthermore, the

production of chitosan from insect exoskeleton have drawn increased attention (Bardawy

and Mohammed, 2015) and in a study by Wanule et al., reveals that chitin and chitosan

can be extracted from American cockroach, Periplaneta americana. One of the most

common specie of cockroach, the American cockroach, according to Pest World.org, are

the largest house-infesting roaches and a major pest in the United States that can pose

severe threat when present at households.

Introduction

As the world becomes more populated, the usage of plastic greatly increases

since it is one of the primary materials that ease the work of individual in various ways.

Not to mention its overall impact for the revolution of society through the lifestyles of

peoples and contribution to innovation. Technology pays great debt to plastic because it

is one of the core materials that lessen the cost of certain products but keeps its durability
intact. Practically speaking, houses including its appliances are approximately 70-80%

made from synthetic materials however; it is also one of the main contributors of

pollution in our world. Plastic wastes is an important issue that every country faces in the

last decades due to its harmful effect in health, overall cleanliness, and even financial

concern that is why no matter how useful plastic can be, the amount of it as wastes

massively disturbs the ecosystem and the government priorities. The motto, REDUCE,

REUSE, RECYCLE might be one of the earliest way to inform people that they must

responsible when using plastics but it did not help the society to eradicate plastic

pollution absolutely and even if people will proceed on erasing plastic in their lives, it

will be a big loss due to its tremendous contribution. Luckily, scientist found a way to

create plastic that mimics the characteristics of petroleum-based or ordinary plastic. Made

from different easily decomposing materials, bioplastic became a trend to a lot of people

and could be the best alternative move to plastic pollution. Several studies were made

about the bioplastic and its sources like starch used or decayed plants and even animals

that possess certain substance capable of creating tangible and durable materials. In this

research, extracted chitosan from Periplaneta americana will be used as the main

component for the production of siomai bioplastic plates.

The purpose of this study is to produce a siomai bioplastic plates that can

mimic the function of the traditional ones and further help the environment through

degradation of its harmful alter-ego due to its drastic increase as solid waste.
Statement of the Problem

1. What are the differences of the chitosan-based Siomai Bioplastic Plates that are
extracted from Periplaneta americana to the ordinary siomai plates?
2. What are the benefits of using Chitosan-based Siomai Bioplastic Plates extracted
from Periplaneta americana in terms of the following fields:
a. Quality of the product
b. Eco-friendliness
c. Cost-efficiency
3. What are the advantages and disadvantages of using Periplaneta americana as
primary source of chitosan for the bioplastic?
4. What are the benefits of Chitosan-based Siomai Bioplastic Plates extracted from
Periplaneta americana for the industrial and commercial sector?

Hypothesis

There are no benefits of Chitosan-based Siomai Bioplastic Plates extracted

from Periplaneta americana for the industrial and commercial sector.

Scope and Delimitation

Producing chitosan based siomai bioplastic plates for the commercial and

industrial sector is what this study aims to establish. In this research, chitosan will be

examined to determine the quality of the product in comparison with other bioplastics

that were made of different substances. Extracted chitosan from Periplaneta americana
that will be processed in the laboratory of the Department of Science and Technology

shall be used in this study.

The classification of other raw materials that can be extracted from

Periplaneta americana will not be used for this research. Other changes in the materials

that will be used shall be disregarded and will not be evaluated in this study. Chitosan

from other arthropods shall not be of any concern in the proceeding of this research thus

limiting to only chitosan from Periplaneta americana due to multiple variation and

irrelevant discrepancy for the experimentation. Statistical records will not be employing

other types of test subjects.

Significance of the Study

This research aims to help the government’s agency that focuses on

environmental rehabilitation, plastic industry, conmercial businesses, product consumers,

and future researchers.

Government Agency. Plastic is one of the main sources of pollution in any

communities and the horrific impact it has proven that government agencies should make

a way to eradicate plastic pollution. However, the usage of synthetic material drastically

increases in the last decade. The biodegradable siomai plastic plate will play a vital role

in replacing its commonly used alter ego that disturbs the environmental stability of any

country.
 Plastic Industry. The success of plastic production have brought prosperity

to people who was behind of their creation but it also brought a mindful conflict to its

industry that could influence the masses and decrease their overall sales. Luckily the

invention of biodegradable siomai plastic plates can be one of the products that the plastic

industry can feature and at the same time contribute to the rehabilitation of pollution.

Commercial Businesses. Siomai businesses are basically the most affected

businesses in this study and the usage of conventional siomai plates are fundamental

materials to ease the services for their costumer however; they are obviously the main

pollutants of siomai plates that is why the biodegradable version of this product can help

them minimize waste.

Product Consumers. Siomai is locally renowned as a street food that is

adored by many people and the increase in number of siomai production also meant tan

increase in number of its container, the siomai plates. However, these containers can only

be used once that’s why they can easily turn into waste. Through the use of biodegradable

siomai plastic plates consumers can dispose the container easily due to its ability to

decompose.

Future Researchers. The evolution of plastic became an integral part of

society for centuries that is why providing a wider scope for the sole purpose of more

refined discovery. The biodegradable siomai plastic plates will be a foundation to more

researches about the use of organic components for the production of one of the most

utilized material of the society.

Definition of Terms

1. Biodegradation- is the decomposition of the bioplastic material made from

chitosan.

2. Bioplastic- the product of the research; also called Bioplastic.

3. Chitosan- the sugar obtained from the Periplaneta americana; the main component

for the production of siomai bioplastic plates.

4. Periplaneta Americana- the primary source chitosan that will be used in this

research.

5. Plastic Waste- solid wastes the serves as pollution in the environment; the

problem that will be fixed in the research.

6. Siomai Plates- the traditional product that will be replicated in this study.
 CHAPTER II

Review of Related Literature

2.1 Bioplastic

According to Gibbens (2018), bioplastic simply refers to plastic made from

plant or other biological material instead of petroleum. Examples of these materials are

corn oil, sugar beet, sugar cane, grass and plants. There has been extensive research and

publicity around the damage traditional plastics are doing to the environment so the fact

that bioplastics are produced from renewable resources and degradable materials means

that can reduce pollution in a very real way (Key Plastic, 2018). Today, bioplastics can be

found in the following market segments: Packaging; Food-services;

Agriculture/horticulture; Consumer electronics; Automotive; Consumer goods and;

household appliances (European Bioplastic, 2019).

 According to European Bioplastic (2016), the global bioplastics production

capacity is set to increase from around 4.2 million tonnes in 2016 to approximately 6.1

million tonnes in 2021. Packaging remains the largest fields of application for bioplastics

with almost 40 percent (1.6 million tonnes) of the total bioplastics market in 2016. The

data also confirms a decisive increase in the uptake of bioplastics materials in many other

sectors, including consumer goods (22 percent, 0.9 million tonnes) and applications in the

automotive and transport sector (14 percent, 0.6 million tonnes) and the construction and

building sector (13 percent, 0.5 million tonnes), where technical performance polymers

are being used.

2.1.1Types of Bioplastic

Based on UrthPact (2018), bioplastic refers to a large category of bio-based

polymers with a variety of unique attributes and applications. European Bioplastic (2016)

classified them into three main groups: Bio-based (or partially bio-based), durable

plastics such as bio-based polyethylene (PE), polyethylene terephthalate (PET) (so-called

drop-in solutions), bio-based technical performance polymers, such as numerous

polyamides (PA), or (partly) bio-based polyurethanes (PUR); Bio-based and

biodegradable, compostable plastics, such as polylactic acid (PLA),

polyhydroxyalkanaoates (PHA), polybutylene succinate (PBS), and starch blends;

Plastics that are based on fossil resources and are biodegradable, such as PBAT and PCL,

but that may well be produced at least partly bio-based in the future.

2.1.3 Composition of Bioplastic

According to Solomonides (2016), the composition of a bioplastic is

comprising of at least one starch, at least one plasticizer, and at least one acid, wherein at

least one starch is between 2 wt. % and 25 wt. %, the at least one plasticizer is between

40 wt. % and 65 wt. %, and the at least one acid is between 1 wt. % to 10 wt. %.

2.1.4 Difference between Bio-based and Biodegradable Plastic

 The term “bioplastics” is actually used for two separate things: bio-based

plastics (plastics made at least partly from biological matter) and biodegradable plastics

(plastics that can be completely broken down by microbes in reasonable timeframe, given

specific conditions). Not all bio-based plastics are biodegradable, and not all

biodegradable plastics are bio-based. And even biodegradable plastics might not

biodegrade in every environment (Krieger, 2019). According to European Bioplastic

(2019), biobased or partially biobased durable plastics, such as biobased or partially

biobased PE, PET or PVC, possess properties, which are identical to their conventional

versions. These bioplastics are technically equivalent to their fossil counterparts; yet, they

help to reduce a product’s carbon footprint. Moreover, they can be mechanically recycled

in existing recycling streams while plastic defined as “biodegradable” is made of

molecules that can break down naturally, but there is no particular timescale specified for

this degradation – under some conditions it can take many years (Thomlinson, 2019),

biodegradable plastics degraded under certain conditions by microorganisms existing in

nature, such as bacteria, molds and algae, and eventually become water or carbon dioxide

and other small molecules into the natural circulation (BAM, 2019)

Biobased does not equal biodegradable. Indeed, even some oil based plastics

can become oxo-degradable or UV-degradable by introducing special additives;

conversely, some polymers of natural origin are not biodegradable. Then there are hybrid

polymeric materials, formed in part by

 natural resources and in part by synthetic polymers. Then, too, there are

polymers with the same molecular chain – and thus identical – that can be produced from

either oil or from renewable raw materials, such as synthetic polyethylene and natural

polyethylene obtained from sugars (Cecchini, 2017).

2.1.5 Biodegradation

Biodegradation is a chemical process during which microorganisms available

in the environment convert materials into natural substances such as water, carbon

dioxide, and compost (no artificial additives required). The biodegradation process

depends on the surrounding environmental conditions (e.g. location or temperature), on

the material, and on the application (European Bioplastic, 2018). A study by Bio-Tec

Environmental clearly stated that biodegradation is nature’s waste management and

recycling system. It breaks down everything from yard waste to crude oil and it is a

natural process necessary to keep our planet clean and healthy. According to Creative

Mechanisms Stuff (2016), the term “bioplastic” represents a plastic substance that is

based (wholly or in part) on organic biomass rather than petroleum. The biodegradability

of a plastic is due to chemical structure of the polymer and not based on the source of

carbon (Darby, 2019).

In the study conducted by Isroi, Supeni, Eris, Cahyaningtyas (2018),

biodegradability of edible bioplastic made from cassava starch by using two natural

inoculums i,e landfill and plantation soil was evaluated. The edible bioplastics used in

this study were bioplastic with and without hydrophobic liquid. Biodegradation test of the

bioplastic was then compared to HDPE plastic. The test was conducted in the glass jar for

60 days. The carbon dioxide generated from the biodegradation process was absorbed by

0.1 N sodium hydroxide and titrated with 0.1 N HCl. There was no carbon dioxide

released from the HDPE plastic during the test. Biodegradation of the bioplastic in

plantation soil was higher than in landfill soil. Biodegradation rate of the bioplastic in

landfill soil was 0.201 mg CO2/day and 0.249 mg CO2/day for bioplastic without and

with hydrophobic liquid, respectively. Biodegradation rate of the bioplastic in plantation

soil was 0.604 mg CO2/day and 0.424 mg CO2/day for bioplastic without and with

hydrophobic liquid, respectively. Complete biodegradation of the bioplastic in landfill

soil was predicted in 431 days and 366 days for bioplastic without and with hydrophobic

liquid, respectively. In another hand, complete biodegradation of the bioplastic in

plantation soil predicted in 151 days and 201 days for bioplastic without and with
hydrophobic liquid, respectively. Microbes population in the soil could be affected the

biodegradation rate of the bioplastics.

In the other hand, biodegradation of the bioplastic composite based on

cellulose from oil palm empty fruit bunches was investigated in the study of Rahman,

Syamsu, & Isroi (2019). Microbes consortium from landfill soil collected from top soil

were used as the inoculums for the biodegradation process. Biodegradation test of the

bioplastic from oil palm empty fruit bunch samples compared with oxodegradation and

conventional plastic samples were conducted in the glass jar. The biodegradation rate was

evaluated from CO2 generated from the biodegradation process and absorbed by 0.1 N

sodium hydroxide solutions. The generated CO2 was titrated with 0.1 N HCl and using

phenol-phthalein (PP) followed by methyl oranges indicator. The results showed that the

highest CO2 production on landfill soil indicating the highest rate of biodegradation was

found on bioplastic from oil palm empty fruit bunch followed by oxodegradable plastic

and conventional plastic. The rate of biodegradation for bioplastic from oil palm empty

fruit bunch, oxodegradable plastic and conventional plastic was 0.067, 0.052 and 0.000

mg CO2/day, respectively.

Both Isroi et al. (2018) and Rahman et al. (2019) concluded that

biodegradation test uses soil as the primary material for the examination of a bioplastic’s

biodegradability. Due to the presence of microbes on the soil, it became one of the main

factors that biodegrades the material. Both research also calculated the CO2 released as

the proper quantitative measure for biodegradation. However, Kemasan (2018) compared
the most essential soil to use for the test between plantation and landfill soil and addition

to that, he also added hydrophobic liquid that became a catalyst as analysed in the data.

Rahman et al. (2019) compared the biodegradation of bioplastic made from cellulose of

oil palm empty fruit bunches, oxodegradation and conventional plastic with the use of

landfill as material of biodegradation. Based on the result, the bioplastic is the fastest

specimen to biodegrade then the oxodegradation plastic and the slowest is the

conventional one.

2.1.5 Chitosan- Based Bioplastic

Scientists at Harvard University’s Wyss Institute for Biologically Inspired

Engineering have developed a new biopolymer that mimics the toughness, durability, and

flexibility of natural insect cuticle. A biological composite with a structure akin to

plywood, insect cuticle consists of layers of the polysaccharide polymer chitin and

protein. The Wyss researchers extracted chitosan sugar from discarded shrimp shells and

engineered a thin, transparent film with a strength equivalent to aluminum at only half the

weight. The new Chitosan Bioplastic is biocompatible, biodegradable, easily molded into

complex shapes, and inexpensive because of the widespread availability of shrimp waste.

Because of these beneficial characteristics, the material is a potential replacement for

conventional petroleum-based plastic in applications ranging from consumer products to

medical uses.

2.2. Periplaneta Americana

 American cockroach, Periplaneta americana (Linnaeus), is the largest of the

common peridomestic cockroaches measuring on average 4 cm in length. It occurs in

buildings throughout Florida, especially in commercial buildings. In the northern United

States, the cockroach is mainly found in steam heat tunnels or large institutional

buildings. It is second only to the German cockroach in abundance (Barbara, 2017). The

American cockroach is perhaps the most cosmopolitan peridomestic pest species.

Together with other closely related Periplaneta species, P. americana is believed to have

spread from tropical Africa to North America and the Caribbean on ships engaged in

slave trading (Mullen & Durden, 2018). Based on the study of Ramazani, Saghafipour &

Vatandoost (2018), the American cockroach lives in hot areas of buildings like the

kitchens, heating rooms, warehouses and sewage systems They usually come out of their

hiding places at night for feeding and other activities. The adult cockroaches are long-

lived and can live for as long as one year or more producing large number of egg

capsules during this period, depending on food availability.

According to Jacob (2013), when indoors, the nymphs and adults are usually

found in dark, warm and moist areas of basements and crawl spaces, and in and around

bathtubs, clothes hampers, floor drains, pipe chases, and sewers. They are also common

around the manholes of sewers, and on the undersides of metal covers over large sump

pumps. In the north, this cockroach is often associated with steam heat tunnels. Where

steam heat tunnels are not found, the American cockroach is restricted primarily to large

institutional buildings. It has also been observed migrating from one building to another

during warm months in the north. Occasionally, the cockroaches infest sanitary landfills
and can survive Pennsylvania winters because of the warmth generated within the piles of

trash. The adults can survive two or three months without food but only about a month

without water.

American cockroaches can become a public health problem due to their

association with human waste and disease and their ability to move from sewers into

homes and commercial establishments (Barbara, 2017). Based on the article of Ray

(2018), the proven health risks of cockroach infestation seem to be predominantly those

of filth, food contamination and allergic asthma rather than direct transmission of disease,

according to an extensive review of public health risks from all kinds of pests conducted

by the World Health Organization. Because many studies have found that cockroaches

feed on human excrement and transfer or excrete pathogens, they have a strong secondary

role in the spread of some diseases. In the other hand, Flowers (2015) specifically studied

the possibility that cockroaches like P. americana can cause allergies. Wherever

cockroaches go, they leave behind traces of their existence. Signs that you have a

cockroach infestation include feces, saliva, and parts of their bodies that they shed or that

fall off. Similar to dust mites, these parts of a cockroach contain specific proteins or

allergens that may cause allergies or can even trigger asthma symptoms.

Flowers (2015) also added that the proteins that cockroaches naturally carry,

there have been tests done that examine the pathogens in their bodies. The scary thing is

that cockroaches can carry some serious diseases. Salmonella Typhi, which causes

Typhoid, has been found in cockroaches. Poliomyelitis, which causes Polio, has also been
found in these insects. They can also cause Dysentery, a disease that causes severe

diarrhea that may include bleeding.

2.2.1. Physiology and Anatomy

Based on PSON Pest Service (2016), each cockroach has eyes, a mouth,

salivary glands, antennae, brain, heart, colon, reproductive system, mid-guts, legs,

esophagus, gastric caecea, fat bodies and malpighian tubules. Most people can recognize

cockroaches instantly. They're brown or black insects that are usually between half an

inch and two inches long (12-50 millimeters), minus their long antennae. Their heads

point downward, almost as if they're built for ramming. Males usually have wings, but

females often don't. Those that do usually have vestigial wings -- small, undeveloped

wings that often don't allow the roach to fly (Wilson, 2020). According to Orkin.com,

cockroach legs are exceptionally sensitive when touched. Their antennae, also known as

feelers, are responsible for their sense of smell. Cockroaches have two small appendages

on their abdomens, known as the cerci, which act as sensors. Cerci give them an

advantage over predators, as they seem to be sensitive to slight air movements around

them.

Based on the study of Fathima (2019), the mouthparts of the cockroach

include labrum, mandibles, maxillae and labium. Labrum or the upper lip is a broad and

roughly rectangular shaped structure. It hangs from the front edge of the head on the

lower side. Hypo pharynx is a tongue like structure in the floor of the mouth. Mandibles
are a pair of hard, strong, large, dark colored structures found one on either side with

jagged inner edges. They move in horizontal motion and crush the food present in-

between them.

Cockroaches are also equipped with salivary glands and an esophagus, which

assist in digestion. At the base of the esophagus, food is temporarily located in the crop.

After entering the stomach of the cockroach, food is broken down by enzymes present

within the gastric caecea and in the middle of the intestines is the mid-gut, which is

responsible for nutrient absorption (PSON Pest Service, 2016). In a study made by

Oyebanji, Soyelu, Bamigbade & Okonji (2014), adult male and female cockroaches,

Periplaneta americana (L.) were assayed for the presence of digestive enzymes in the

fore-, mid- and hindgut regions. Activities of α-amylase, β-amylase, γ-amylase,

proteinase and lipase were detected in the three gut regions except for the absence of γ-

amylase and lipase in female hindgut. The presence of these enzymes partly explains the

polyphagous feeding habit of P. americana, enabling the insect species to digest a wide

variety of food substances. In some instances, significant differences in enzyme activities

were observed between sexes and among gut regions. Generally, enzyme activity was

highest in the midgut followed by the fore- and hindgut in descending order. Despite this

trend, a considerable level of proteinase and male lipase was observed in the hindguts

suggesting that it might be necessary to give extra attention to hindgut activities in future

studies.
 Tanner (2020) stated that male cockroaches have colorless blood, while the

female cockroaches may occasionally have orange blood. This is due to the absence of

hemoglobin in their blood (the same cell that makes human blood red). Human beings

use hemoglobin to carry oxygen, but cockroaches do not breathe the way we do.

Cockroaches use a series of tubes called tracheae for breathing (and exhaling) and in fact

they can breathe even in absence of a head. Cockroaches, like other insects, have an open

circulatory system and their blood is also known as hemolymph (or haemolymph). It

flows freely inside the body, touching all the internal organs and tissues. About 90% of

this blood is watery fluid and the remaining 10% is made up of hemocytes. The oxygen is

delivered by the tracheal system and not the circulatory system in a cockroaches (or most

of the other insects).

2.2.2. Taxonomy

Accordimg to PSON Pest Service (2016), under the Kingdom Animalia,

cockroaches belong to the Phylum Arthropoda, Class Insecta, and Order Blattodea. The

Order name is derived from the Greek blatta. Some of the scientific names of the
different species of cockroaches are the American cockroach (Periplaneta americana), the

Florida woods cockroach (Eurycotis floridana), the Oriental cockroach (Blatta orientalis),

the German cockroach (Blattella germanica), the Asian cockroach (Blattellaasahinai), the

Surinam cockroach (Pycnoscelus surinamensis), the brown-banded cockroach (Supella

longipalpa), the Australian cockroach (Periplaneta australasiae), the smoky brown

cockroach (Periplaneta fuliginosa), the Pennsylvania woods cockroach (Parcoblatta

pennsylvanica), the brown cockroach (Periplaneta brunnea), and the Madagascar hissing

cockroach (Gromphadorhina portentosa). Based on the ITIS Report (nd), the taxonomic

Hierarchy of P. americana is as follows:

Kingdom Animalia
Subkingdom Bilateria
Infrakingdom Protostomia
Superphylum Ecdysozoa
Phylum Arthropoda
Subphylum Hexapoda
Class Insecta
Subclass Pterygota
Infraclass Neoptera
Superorder Polyneoptera
Order Blattodea
Superfamily Blattoidea
Family Blattidae
Subfamily Blattinae
Genus Periplaneta
 Species Periplaneta americana (Linnaeus, 1758)

2.2.3. Extracting Chitosan from P. americana

Chitosan from the exoskeleton of the cockroach (Periplaneta americana L.)

were extracted using the following chemical treatment with an acid and alkali: For chitin

extraction, 2 N HCl and 1.25 N NaOH solutions were used to achieve demineralization

and deproteinization, respectively. To obtain chitosan, chitin particles from cockroaches

were boiled at different NaOH concentrations and temperatures. The relative average

yields of chitin and chitosan from cockroaches were 3.36% and 2.08%, respectively, on a

fresh-weight basis. The relative average yield of chitin and chitosan was 12.17% and

7.28%, respectively, on a dry-weight basis. The deacetylation percentage of chitosan

from exoskeletons of cockroaches were 90.03%, 91.86%, and 89.4% in Procedures B, C,

and D, respectively. The viscosity of chitosan extracted from exoskeletons in Procedures

B, C, and D was 48.0, 69.6, and 36.0 cP, respectively. The molecular weight of the

obtained chitosan was 230,300 Da in Procedure C and 212,100 Da in Procedure D. Our

results indicate that exoskeletons of cockroaches may serve as a source of chitin and

chitosan for livestock and cultured fish industry and for preparation of elicitors for

improvement of resistance to phytopathogens in agriculture (Wanule & Balkhande,

2019).
2.3. Chitosan

Innace & Di Maio (2014), defined chitosan (CS) as a biodegradable cationic

aminopolysaccharide derived from chitin (N-deacetylated derivative), the second most

abundant polysaccharide after cellulose. Chitosan is an aminated polysaccharide

abundant in nature. Its chemical characteristics provide chitosan with a unique set of

functional properties. The main characteristics of chitosan are its degree of acetylation

and molecular weight. These have a determining effect on the chitosan functional

properties, from its solubility and materials-forming capacity to biodegradability and

diverse bioactive attributes. The great potential of use of chitosan in diverse fields has

long been recognized. The biological related properties of chitosan make it particularly

suitable for applications related with agricultural, food, and environmental engineering

uses.

Chitin is generally found as ordered crystalline micro fibrils in the structural

component of crustaceans and insects, and is also found in the cells of fungi and

microorganisms (Verma & Fortunati, 2019). Currently, alternative sources are used such

as seafood waste (Yadav, Goswami, Paritosh, Kumar, Pareek, Vivekand, 2019) and larval

exoskeleton (Hahn, Roth, Ji, Schmitt, & Zibek 2019).

Chitosan and chitin have a variety of uses and CuanTec (2018) made a list of

significant sectors where they can contribute:

In agriculture chitosan can be used to be a fertilizer and pesticide as a

clarifier and a water filtering aid, both chitin and chitosan both bond easily with metals
and micro contaminants under water, while being non-allergenic, non-toxic and

biodegradable, making the water clearer. It is usable in areas like lakes, since they are

totally natural, making them viable alternatives to chemicals water treatments.

They are great for the creation of Scaffold in tissue engineering because of:

their high porosity, biodegradability, non-toxicity, and high biocompatibility. Chitosan

are also great for wound healing, thanks to their blood blotting ability and antimicrobial

activities.

Chitin is used in cosmetic because of its hypoallergenic characteristic,

making it highly tolerant. It is used as a moisturizing agent for two reasons: it supplies

water; and retain water, avoiding dehydration. It is used in hairspray: to increase stiffness,

make curls last longer. Chitin can be used as absorbable surgical suture to replace

synthetic polymer currently having a monopoly on the area.

Bibliography

Barabara, K.A. (2017). American Cockroach, Periplaneta americana (Linnaeus) (Insecta:

Blattodea: Blattidae). University of Florida: IFAS Extension. Retrieved

from: https://edis.ifas.ufl.edu/in298
Cecchini, C. (2017). Bioplastics made from upcycled food waste. Prospects for their use

in the field of design. The Design Journal. Retrieved from:

https://www.tandfonline.com/doi/abs/10.1080/14606925.2017.1352684

Creative Mechanisms Stuff. (2016). Everything You Need to Know About Bioplastics.

Retrieved from: https://www.creativemechanisms.com/blog/everything-you-

need-to-know-about-bioplastics

CuanTec.(2018). Applications and uses of Chitosan and Chitin. Retrieved from:

https://www.cuantec.com/blog-and-news/blog/108-applications-and-uses-

of-chitosan-and-chitin-2

Darby, D. (2019). Bioplastcs and Biodegradability. North East Recycling Council.

Retrieved from: https://nerc.org/news-and-updates/blog/nerc-

blog/2019/08/20/bioplastics-and-biodegradability?

fbclid=IwAR0h3INoB3KTQ1P2pmDo25YolJDtMlk1MSAjjlIdUfvkYk45B

QUrcXfrh8I

European Bioplastic. (2016). What types of bioplastics do exist and what properties do

they have? Retrieved from: https://www.european-bioplastics.org/faq-

items/what-types-of-bioplastics-do-exist-and-what-properties-do-they-have/

European Bioplastic. (2018). What are bioplastics? Retrieved from:

https://www.european-bioplastics.org/bioplastics/
European Bioplastic. (2019). Applications for bioplastics. Retrieved from:

https://www.european-bioplastics.org/market/applications-sectors/

Flowers, J. (2015). How Cockroaches Can Make You Sick (And What You Can Do

About It). Retrieved from: https://learn.allergyandair.com/cockroach-

allergies/

Gibbens, S. (2018). What you need to know about plant-based plastics. National

Geographic. Retrieved from:

https://www.nationalgeographic.com/environment/2018/11/are-bioplastics-

made-from-plants-better-for-environment-ocean-plastic/

Hahn, T., Roth, A., Ji, R., Schmitt, E. & Zibek, S. (2019). Chitosan Production with

Larval Exoskeletons derived from the Insect Protein Production. Retrieved

from:

https://www.sciencedirect.com/science/article/pii/S0168165619309514

Isroi, I., Supeni, G., Eris, D. & Cahyaningtyas, A. (2018). Biodegradability of Cassava

Edible Bioplastic in Landfill Soil and Plantation Soil. Retrieved from:

http://ejournal.kemenperin.go.id/jkk/article/view/3596

ITIS Report. (nd.). Periplaneta americana (Linnaeus, 1758). Retrieved from:

https://www.itis.gov/servlet/SingleRpt/SingleRpt?

search_topic=TSN&search_value=102406#null
Jacobs, S. (2013). American Cockroaches. Penn State Extension. Retrieved from:

https://extension.psu.edu/american-cockroaches

Iannace, S. & Di Maio, E. (2014). Biomedical Foams for Tissue Engineering

Applications. Retrieved from:

https://www.sciencedirect.com/science/article/pii/B9780857096968500061#

Key Plastic. (2018). 6 Common Applications of Bioplastics, Retrieved from:

http://www.keyplastics.ie/6-common-applications-bioplastics/

Krieger, A. (2019). Are Bioplastic better for the Environment than Conventional Plastics.

Retrieved from: https://ensia.com/features/bioplastics-bio-based-

biodegradable-environment/

Mullen, G. & Durden, L. (2018). Medical Veterinary Entomology- 3rd Edition.

PSON Pest Service. (2016). Cockroach Anatomy. Retrieved from:

https://www.psonpest.com/%E0%B8%A3%E0%B8%B2%E0%B8%A2%E

0%B8%A5%E0%B8%B0%E0%B9%80%E0%B8%AD

%E0%B8%B5%E0%B8%A2%E0%B8%94/583e45a668cfd90001f6003e/C

ockroach%20Anatomy/5843d15268cfd90001f667c3/langEN?

fbclid=IwAR1L76JrcKe_pVXRkIweBlqQ6uz5jwuKV74D01Ac3AOrUD42

f8_SSCoQT8o

Rahman, A., Syamsu, K. & Isroi, I. (2019). Biodegradability of Bioplastic in Natural

Environment. Retrieved from:

 https://www.researchgate.net/publication/334399882_BIODEGRADABILI

TY_OF_BIOPLASTIC_IN_NATURAL_ENVIRONMENT

Ramazani, A., Saghafipour, A. & Vatandoost, H. (2018). Control of American Cockroach

(Periplaneta americana) in Municipal Sewage Disposal System, Central

Iran. Retrieved from:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6091800/

Ray, C. (2018). Do Cockroaches Carry Diseases? New York Times. Retrieved from:

https://www.nytimes.com/2018/08/03/science/cockroach-diseases.html

Solomonides, E. G. (2016). Biodegradable Bioplastic Compositions and Methods of

Making and Using the Same. United States Patent Application

20160369085. Retrieved from:

http://www.freepatentsonline.com/y2016/0369085.html

Thomlinson, I. (2019). When biodegradable plastic is not biodegradable. Retrieved from:

http://theconversation.com/when-biodegradable-plastic-is-not-

biodegradable-116368

UrthPact. (2018). Basics of Bioplastic. Retrieved from:

https://www.urthpact.com/tag/what-are-bioplastics-made-of/

Verma, D., Fortunati, E. (2019). Biomass, Biopolymer-Based Materials, and Bioenergy.

Retrieved from:
 https://www.sciencedirect.com/book/9780081024263/biomass-biopolymer-

based-materials-and-bioenergy

Wanule, D. & Balkhande, J.V. (2019). Extraction and FTIR Analysis of Chitosan from

American cockroach, Periplaneta Americana. Retrieved from:

https://www.researchgate.net/publication/269285580_Extraction_and_FTIR

_Analysis_of_Chitosan_from_American_cockroach_Periplaneta_americana

Wyss Institute. (2011). Inspired by Insect Cuticle, Wyss Researchers Develop Low-Cost

Material with Exceptional Strength and Toughness. Retrieved from:

https://wyss.harvard.edu/news/inspired-by-insect-cuticle-wyss-researchers-

develop-low-cost-material-with-exceptional-strength-and-toughness/

Yadav, M., Goswami, P., Paritosh, K., Kumar, M., Pareek, N. & Vivekanand, V. (2019).

Seafood waste: a source for preparation of commercially employable

chitin/chitosan materials. Retrieved from:

https://link.springer.com/article/10.1186/s40643-019-0243-y
 CHAPTER III

Methodology

3.1 Experimental Design

The purpose of this study is to create a cheap, high-quality but eco-friendly

bioplastic product that can replicate and eventually, replace the ordinary siomai plates

that are widely used in the commercial sector. The objectives of this experiment are to

determine the relationship of the amount of chitosan extracted from Periplaneta

americana incorporated in the bioplastic with its breaking strength, solvent uptake and

biodegradation. This will be proven in different ratios of chitosan that will set significant

differences between treatments A, B, C, D, and E (where E is the original siomai plates).

The ordinary bioplastic plate is locally labelled as an infamous solid pollutant that are

found in almost every part of the community that is why substituting a bioplastic that will

not only mimic the traditional use of the old ones, products made from natural,

biodegrading, and eco-friendly substance will lessen the amount of solid waste in the

area. Chitosan are the latest but most efficient substance that can copy the characteristics

of plastic, so far.
3.2 Materials and Equipments

The materials that will be used in this study are chitosan that are extracted from

Periplaneta americana tested in DOST-PH, arrow-root starch, castor oil, ethanol, sodium

hydroxide pH 10, deionized water, hydrochloric acid pH 5, nutrients agar (NA), and

aluminum foil. The tools and equipments that will used in this research are glassware,

hotplate, analytical scales, petri dish, tensile test equipment (IK Force Tester MCT-2150),

needle, oven, spirit burner, laminar flow, and autoclave.

3.3 Methodological Framework

3.3.1 Preparation of material

Gathering of Materials Measuring of Ratio

Materials will be gathered, cleaned, and sorted in preparation for use during the

experiment. The materials are then measured and labelled to the amount needed in each

treatment. Chitosan powder is measured. Other ingredients are properly labelled and

measured as well
3.3.2 Extraction of Chitosan

Adult Periplaneta americana of sex will be procured from IDTI Laboratory

Animal House for the extraction of chitosan. They will be starved for 48hrs before

sacrificing and the sacrificed cockroaches will be dried in oven at 65°C for 48 hrs. The

samples will be powedered to extract the chitosan as per the standard method; 10 grams

of powder will be treated with 4% of NaOH for 1hr in order to dissolve protein and sugar

to isolate crude chitin. Samples will be boiled in 4% NaOH on hot plate and then it

allows cooling for 30 minutes at room temperature. After cooling each sample, it will be

washed thoroughly with deionized water thrice. Supernatant should be discarded and

residue will be demineralized by treating it with 20 ml of 1% HCl for 24 hrs. The

demineralized samples will then be washed with deionized water. After washing the

residue sample will be treated with 50 ml of 2% NaOH solution for 1 hr. It will

decompose albumin into water soluble amino acids. The supernatant should be discarded

and remaining chitin will be washed with deionized water. The chitin will be converted

into chitosan by process of deacetylation .The process will carried out by adding 50 ml of

50% NaOH to all samples and then boiled at 100°C for 2hr. on the hot plate. Samples

will then be cooled for 30 min at room temperature. After cooling, sample will be washed

continuously with 50 % of NaOH and filtered in order to retain the solid mater. This solid

mater is further washed thrice with deionized water. This solid mater is going to be dried

in oven at 120° C for 24hrs.

3.3.3 Bioplastic Synthesis

Bioplastic synthesis will be performed by weighing chitosan with varying

predetermined mass. Then the chitosan will dissolved into 5% acetic acid while stirring

with a magnetic stirrer. The same thing will also be applied on arrow-root starch until all

dissolves. Both solutions will be mixed in a beaker and added with 15% of castor oil.

After the sample begins to form the gel, the sample is poured into the platter covered with

aluminium foil and dried over the hotplate at 75 ° C until all the solvents evaporate and

plastic films are obtained.

3.3.4 Tensile test.

Bioplastic samples will be cut to lengths of 4 cm and 1 cm wide. The

measurement condition is a tensile velocity of 100 mm / min with a maximum load of

500 N. The sample will be clamped on a tensile test apparatus, and runs the apparatus

according to a predetermined condition until the sample is disconnected. Repeated

procedure above for each sample will be done 3 times. The magnitude of the breaking

strength of a material depends on the amount of loading applied and the cross section of

the material itself. Recorded tensile test results are presented in the form of tables

between tensile strength and extension.

3.3.5 Solvent uptake test.

This test will be based on the method performed by Pimpan, et al., 2001. Plastic

will be cut to the size of 1.0 cm x 1.0 cm then weighed with an analytical balance sheet.

The plastic is put into a 10 ml beaker filled with 5 ml solvent, then sterilized in room

temperature. Every minute, the plastic will be taken; the solvent on the plastic surface is

wiped with a tissue, and then weighed. The absorption capacity of the solvent is

calculated.

3.3.6 Biodegradation test.

Bioplastic samples will be cut to 1 x 1 cm size then sterilized using UV light in

the laminar flow for 20 minutes. Sterilized bioplastic samples will be incorporated into

the biodegradation medium then incubated for 5, 10, 15 and 20 days.

3.3.7 Shaping of Bioplastic

After doing all the test for the quality of the bioplastic, a new sample of the

solution will be shaped using an aluminium covered 18 cm x 5.5 cm x 1.5 cm siomai

plastic plates that will be placed on a hot plate at 75°C until all solvent are evaporated

and the product is acquired.

3.4 Statistical Analysis

Collection of Data

Statistical Test
(Line Regression)

Tabulate Results

The data confined will be interpreted using a Line Regression, according to

Devault (2020), linear regression models are used to show or predict the relationship

between two variables or factors. The factor that is being predicted (the factor that the

equation solves for) is called the dependent variable. The factors that are used to predict

the value of the dependent variable are called the independent variables. In linear

regression, each observation consists of two values. One value is for the dependent

variable and one value is for the independent variable. In this simple model, a straight

line approximates the relationship between the dependent variable and the independent

variable

In this study, the researcher will be using chitosan that is extracted from

Periplaneta Americana with varying measurements. The quantity of chitosan will be

correlated with the result of tensile test, solvent up take test, and biodegration test

wherein the independent variable, chitosan will be the main component tested to which in

may influence the quality of biodegradable siomai plastic plates.