INTRODUCTION

Background of The Study

The depletion of fossil fuels has caused a significant increase in the oil prices since last

two decades. The energy crises are distinct threat to the sustainable development in third world

countries. Fuel prices in the Philippines are among the most expensive in Southeast Asia

(Philippine Daily Inquirer, 2018) and the fossil fuels energy consumption in the Philippines was

reported at 61.99% in 2014, according to the World Bank collection of development indicators.

Since fuels aren’t getting any cheaper, some people can’t even afford it. The global economy can

no longer depend on fossil fuels and non-renewable carbon and with reference to current energy

scenario, scientific community needs to pay special attention on alternative or renewable fuels,

such as biofuels as the feedstock is widely available thus in the last two decades, global research

programs on alternative energy have been directed towards discovering new and sustainable

energy sources. To settle those setbacks, humans need to find more abundant and renewable

source of fuel that is cost effective and an alternative source that could keep the world running

on its wheel.

The energy system is very unstable because the energy resources on earth are limited, and

humans have utilized the conventional technologies to fulfill our needs. Consumption of energy

by humans increased gradually as human needs also increase. Energy resources mainly include

fossil fuels, but research shows that these fuels source will be consumed completely in

approximately 20-25 years. This could cause major problems around the world. As a result, we

need to discover and produce an alternative source of fuel that could keep the world running on

its wheel. One possibility is ethanol.

1
 Ethanol is also referred as ethyl alcohol, grain alcohol or ETOH, that has a characteristic

of clear liquid with an agreeable odor. Ethanol can be made basically in two methods which are

by fermenting and distilling simple sugars. Producers of ethanol mostly benefit upon agricultural

setting which is most commonly practiced in tropical countries. Most fuel ethanol produced in

the U.S. is derived from corn. Latest figures indicate that almost 40 percent of the U.S. corn crop

is dedicated to ethanol production (Red Field Energy, LLC, 2018). One of the most top

producers of ethanol of Brazil using sugar being one of their feedstock. Since it is produced from

crops or plants that harness the power of the sun, ethanol is considered a renewable fuel.

According to Statistics MRC, the global Bioethanol market is valued at $48.7 billion in 2016 and

is expected to grow at a CAGR of 6.4% to reach $75.5 billion by 2023. The global market for

bioethanol derives much of its growth from the increasing demand for renewable energy sources

(Herald Keeper via COMTEX, 2015). North America has been leading the global market for the

source of bioethanol. The growing usage of these products in the transportation industry and

growing government & environment regulations are the factors anticipated to fuel the demand

for bioethanol in North America over the future, ensuring its popularity. While the Latins and

Asians are able to witness the strong growth due to the availability of arable land in India, China

and Brazil.

Ethanol is currently produced from the easily fermented sugar and starch in grain and

food processing wastes. There will be many benefits by connecting the established corn ethanol

industry with the emerging technologies that produce ethanol from agricultural wastes and other

types of biomass. Currently, the United States produces approximately three billion gallons of

ethanol from corn annually (Potera, 2004). The sugar beets and sugar cane are very good

examples of these feed stocks that contain the sugar. Bioethanol is mainly produced by the sugar

2
fermentation process. Saccharomyces cerevisiae are microorganisms that cannot be seen with the

naked eye, they convert carbon sources (Sugar) to bioethanol during fermentation process,

thereby meeting energy demand (Ojewumi, 2017). In the breweries, Saccharomyces cerevisiae is

used in alcohol production and this occurs by converting sugar to energy. Saccharomyces

cerevisiae is also used in the bakeries for raising dough. Other enzyme uses dextrin to make

glucose especially during saccarification. As compared to Saccharomyces cerevisiae,

Zymomonas mobilis can also ferment but it is limited to glucose, fructose and sucrose. (Cardona,

2008).

Ethanol is blended in all the gasolines in the country today especially in the Philippines.

The Biofuels Act of 2006 requires all companies in the Philippines to mix in 10% of ethanol in

all their gasoline. Ethanol (or E10), which is made from sweet sorghum, corn, and cassava, also

uses our abundant sugarcanes (primarily in Mindanao, Luzon, Negros and Panay) for its

production (Carmudi Philippines, 2018). There are five reasons why they are included in our

gasoline today because of environment-friendly factor, the support sugar and other crop farmers

in the country, it help lessen the dependence on other countries for fuel, it is safe to use in cars;

and cost-effective.

According to Ramkumar Nair of University of Boras (2017), they have been verifying a

process that they hope will work in an industrial scale, when it comes to using existing ethanol

factories. Through that process, the industry can become more sustainable and use agricultural or

industrial waste for the production of bioethanol. Bioethanol is used for fuel for ethanol cars,

among other things. Usually, wheat, sugar canes, or corn are used for ethanol production. In

Sweden, wheat is the most common. "But these are crops that could be used as human nutrition,"

Ramkumar Nair says. “It is more sustainable if we could use waste to create fuel, and this is

3
something we have been working on in several projects here at the University of Borås.”

Ramkumar Nair tells us that these research and pilots project where waste is used are called the

second generation ethanol processes, whereas the current industrial production is called the first

generation ethanol process. Ramkumar has now verified a process that integrates the first and the

second generation ethanol processes.

Thus, the researchers came up with an idea of using coconut (Cocos nucifera) shell as an

alternative source of second generation bioethanol by using Saccharomyces cerevisiae as its

reagent. The coconut shell has a high calorific value of 20.8MJ/kg and can be used to produce

steam, energy-rich gases, bio-oil, biochar and etc. Coconut shell contains lower ash content, high

volatile matter and mostly available in triopical countries which is at a cheap cost. It is to be

noted that coconut shell and coconut husk are solid fuels that are usually prominent in making

fuel due to their potential characteristics (Salman Zafar, 2020). By fermenting and distilling the

ground coconut shell to produce an alternative source of bioethanol will be a great help to solve

the problem because coconut is renewable, abundant, and locally available.

4
Statement of The Problem

This study aims to utilize shell of coconut (Cocos nucifera) to find out if it is a possible

source of bioethanol using Saccharomyces cerevisiae.

This study aims to find out:

1. Is there any significant difference on the amount of ethanol produced from coconut

shell (Cocos nucifera) using Saccharomyces cerevisiae based on the fermentation

time?

2. Is there any significant difference between the commercial ethanol and the ethanol

produced from coconut shell (Cocos nucifera) in terms of:

a. Combustibility

b. Color of flame

3. Is coconut shell (Cocos nucifera) effective in producing ethanol?

Null Hypotheses

Ho : There is no significant difference on the amount of ethanol produced using coconut shell

(Cocos nucifera) using Saccharomyces cerevisiae based on the fermentation time.

Ho : There is no significant difference between the commercial ethanol and the ethanol produced

from coconut shell (Cocos nucifera) in terms of:

a. Combustibility

b. Color of flame

Ho : Coconut shell (Cocos nucifera) is not effective in producing ethanol.

5
Significance of The Study

This study will be conducted to derive bioethanol from coconut shell. It aims to reduce

the use of commercial ethanol which causes pollution and loss of non-renewable resources. It

presents a natural, alternative, cheaper, human and environmentally friendly bioethanol. Thus,

the making of ethanol makes transportation cheaper than before. This research can also give

knowledge to the future researchers in producing more bioethanol in the future.

The significance of this study is when the researchers use the shell of coconut for

producing a bioethanol gas; the researchers are not just helping the environment to be clean, we

are also relating the matter of reusing the unused things that we can see in our environment.

Coconuts are commonly seen in our country, and sometimes, some parts of it are wasted. Ethanol

is much less likely to catch fire and less possibility to explode in case of fuel leakage. It has also

low green house gases emission.We can avoid fires that may occur while using bioethanol gas

and it is less expensive than the commonly used gas. This is why the idea of using coconut shell

for bioethanol production came up on the researchers.

Scope and Delimitation

This study focuses on the utilization of waste materials produced into a useful one. This

study will be conducted and use coconut shell as an alternative source of bioethanol for industrial

uses. The coconut shells will be collected in the the vicinity of Concepcion, Tarlac and the

experimentation will be conducted at St. Jude Village, Concepcion, Tarlac and the school

laboratory of Benigno S. Aquino National High School, Senior High School Department, San

Nicolas Poblacion, Concepcion, Tarlac. The coconut shells will undergo pretreatment, where the

recalcitrant structure of lignocellulose is disrupted resulting in breakage of lignin sheath,

6
degradation of hemicellulose and reduction in crystallinity and degree of polymerization of

cellulose (Loow et al., 2015; Chen et al., 2017) and alkaline pretreatment which is a widely

studied chemical pretreatment method which is based on the solubilization of lignin in the

alkaline solution. Among the various alkaline reagents used commonly for alkaline pretreatment,

sodium hydroxide was found to be the most effective (Kim et al., 2016) that’s why it was the

alkaline reagent used in this research. The samples will also undergo acid hydrolysis in order to

break down the cellulose and hemicellulose polymers in lignocellulosic biomass to form

individual sugar molecules which can be fermented into ethanol. Fermentation was also

conducted where yeast (Saccharomyces cerevisiae) is added to the solution, and then fermented

for days (5, 7, and 9 days). The yeast (Saccharomyces cerevisiae) contains an enzyme called

invertase, which acts as a catalyst and helps to convert the sucrose sugars into glucose and

fructose. The last procedure is distillation which is the most dominant and recognized industrial

purification technique of ethanol. Distillation setup will be used in order to produce the

bioethanol by distillation process conducted in the laboratory.

The fermented samples will be compared in terms of the volume of ethanol produced

during the distillation process to determine any significant difference in terms of the

fermentation time on ethanol production. The fermented samples will be tested in terms of their

color and combustibility and were compared to the commercial one. The color of the flame,

smoke and the duration of the flame between the fermented samples will be observed. The

commercial ethanol, the controlled variable is to compare the results of the alternative ethanol.

The samples will undergo combustion test to measure how easily the samples ignite, how

quickly they burn and how they react when burned.

7
 The data to be gathered during the experimentation would be presented using series of

tables which will indicate any significant difference on the amount of volume produced during

the ethanol production based on the fermentation time of the different fermented samples and to

compare the produced ethanol from the commercial one. To interpret the data effectively, the

researchers will employ the Analysis of Variance (ANOVA).

8
REVIEW OF RELATED LITERATURE

Bioethanol

Alternatives to petroleum-derived fuels are being sought in order to reduce the world's

dependence on non-renewable resources. The most common renewable fuel today is ethanol

derived from corn grain (starch) and sugar cane (sucrose). It is expected that there will be limits

to the supply of these raw materials in the near future, therefore lignocellulosic biomass is seen

as an attractive feedstock for future supplies of ethanol. However, there are technical and

economical impediments to the development of a commercial processes utilizing biomass.

Technologies are being developed that will allow cost-effective conversion of biomass into fuels

and chemicals. These technologies include low-cost thermochemical pretreatment, highly

effective cellulases and hemicellulases and efficient and robust fermentative microorganisms.

Many advances have been made over the past few years that make commercialization more

promising. (Gray et al., 2006).

According to Ramkumar Nair of University of Boras (2017), they have been verifying a

process that they hope will work in an industrial scale, when it comes to using existing ethanol

factories. Through that process, the industry can become more sustainable and use agricultural or

industrial waste for the production of bioethanol. Bioethanol is used for fuel for ethanol cars,

among other things. Usually, wheat, sugar canes, or corn are used for ethanol production. In

Sweden, wheat is the most common. "But these are crops that could be used as human nutrition,"

Ramkumar Nair says. “It is more sustainable if we could use waste to create fuel, and this is

something we have been working on in several projects here at the University of Borås.”

Ramkumar Nair tells us that these research and pilots project where waste is used are called the

second generation ethanol processes, whereas the current industrial production is called the first

9
generation ethanol process. Ramkumar has now verified a process that integrates the first and the

second generation ethanol processes.

Ethanol

Also referred to as ethyl alcohol, grain alcohol or ETOH, ethanol is a clear liquid with an

agreeable odor. Ethanol is made by fermenting and distilling simple sugars. Today, ethanol is a

fuel produced from crops such as corn, grain sorghum, wheat, sugar and other agricultural

feedstocks. Most fuel ethanol produced in the U.S. is derived from corn. Latest figures indicate

that almost 40 percent of the U.S. corn crop is dedicated to ethanol production. In Brazil, the

world’s top producer of ethanol, sugar is the primary feedstock. Since it is produced from crops

or plants that harness the power of the sun, ethanol is considered a renewable fuel. Pure fuel

ethanol contains chemical properties identical to that of other grain alcohol. As such, it must be

denatured or made unfit for human consumption. In the U.S., the most common denaturant is

gasoline. (Red Field Energy, LLC, 2018).

Fermentation

Fermentation is one important biological technology that can synthesize various valuable

compounds to satisfy the requirement of chemical industry, energy, food, etc. (Bio resource

Technology, 2017). Yeast and certain bacteria perform ethanol fermentation where pyruvate

(from glucose metabolism) is broken into ethanol and carbon dioxide. The net chemical

equation for the production of ethanol from glucose is:

C6H12O6 (glucose) → 2 C2H5OH (ethanol) + 2 CO2 (carbon dioxide)

10
 Ethanol fermentation has used the production of beer, wine, and bread. It's worth noting

that fermentation in the presence of high levels of pectin results in the production of small

amounts of methanol, which is toxic when consumed. (Anne Marie Helmenstine, 2019)

Hydrolysis

In its simplest definition, hydrolysis is a chemical reaction in which water is used to

break down the bonds of a particular substance. In biotechnology and as far as living organisms

are concerned, these substances are often polymers (simply put, many similar molecules can that

join together). Hydrolysis can also be thought of as the exact opposite reaction to condensation,

which is the process whereby two molecules combine to form one larger molecule. The end

result of this reaction is that the larger molecule ejects a water molecule. (Theresa Philips, 2019).

Coconut Palm (Cocos nucifera)

The coconut is the most cultivated nut in the world and a very important commercial

crop in many tropical countries. The nut meat, coconut oil from the dried meat and milk are

used for drinks, cooking, cosmetics and numerous other types of products. Unopened flowers can

potentially produce coconut molasses and the hard-grained shell of the nut is carved into

practical utensils and ornamental objects. Coconut palms are propagated only from seed. The

seeds, or coconut nuts, are ready for planting when they make a sloshing sound upon shaking.

Following soaking in water for two or three days, the nut is placed on its side or with the pointed

end down in a well-drained site or container with well-drained growing medium and covered

about halfway with sand or mulch. Germination occurs best with regular moisture and

temperatures between 90 and 100 degrees Fahrenheit and takes three to six months. Coconut

11
palms grow rapidly and can begin to bear fruit after as few as five years. (Angela Ryczkowski,

2018)

Coconut Shell

The coconut shell has a high calorific value of 20.8MJ/kg and can be used to produce

steam, energy-rich gases, bio-oil, biochar etc. It is to be noted that coconut shell and coconut

husk are solid fuels that are usually prominent in making fuel due to their potential

characteristics. Coconut shell contains lower ash content, high volatile matter and mostly

available in triopical countries which is at a cheap cost. The higher fixed carbon content leads to

the production to a high-quality solid residue which can be used as activated carbon in

wastewater treatment. Coconut shell is more popular in tropical countries which the production

of coconut is plenty. (Salman Zafar, 2020)

12
RELATED STUDIES

Due to the loss of fuel resources the prices of fuels are continuously rising. Fuel prices in

the Philippines are among the most expensive in Southeast Asia (Philippine Daily Inquirer,

2018). And the fossil fuels energy consumption in the Philippines was reported at 61.99% in

2014, according to the World Bank collection of development indicators. Since fuels aren’t

getting any cheaper, some people can’t even afford it. To settle those setbacks, humans need to

find more abundant and renewable source of fuel that is cost effective and an alternative source

that could keep the world running on its wheel.

Bioethanol

Bioethanol is known as ethylic alcohol, it is typically produced from the fermentation of

various feedstock that contain mainly fermentable sugar or rich in carbohydrates. Its molecular

formula is CH3CH2OH same organic compound that is used in alcoholic beverages. Bioethanol is

commonly used liquid biofuel in the world. Raw feedstock usually has lignin content. It can be

produced from fermentation vegetable waste.

Ethanol

Ethanol is often used as topical disinfectant. It pharmaceutical preparations is used as

solvent and preservatives as well as serving as the primary ingredient in alcoholic beverages.

Indeed, ethanol has widespread use as a solvent of substances intended for human contact or

consumption, including scents, flavorings, coloring, and medicines. Ethanol is the leading liquid

biofuel used for transportation. First-generation ethanol has a simple production process using

sugar or grain as raw material (sugarcane juice in Brazil and corn in the USA and EU, for

13
example), while 2G ethanol (bioethanol) has more complex steps of production and uses

lignocellulosic material as a substrate (Dos Santos et al., 2017)

Fermentation

Fermentation is a metabolic process that converts sugar to acid derivatives, various gases,

or alcohol. Another definition is that fermentation is the chemical process by which molecules

such as glucose (C6H12O6) are decomposed anaerobically. The process can involve complete

decomposition of the glucose to carbon dioxide and water (+ energy) or can be adapted to

produce ethanol (ethyl alcohol + energy). Fermentation offers a pathway for the use

of biomass as a means of producing organic chemicals either directly or organic chemicals as

feedstocks for the production of other chemicals. (Speight, 2017).

Saccharomyces cerevisiae are microorganisms that cannot be seen with the naked eye,

they convert carbon sources (Sugar) to bioethanol during fermentation process, thereby meeting

energy demand (Ojewumi, 2017). Saccharomyces cerevisiae has traditionally been used to

produce alcohol in brewing and wine industries (Azhar et al., 2017). In the breweries,

Saccharomyces cerevisiae is used in alcohol production and this occurs by converting sugar to

energy. . This yeast produces high yields of ethanol with high productivity (Lama et al., 2012).

Saccharomyces cerevisiae is also used in the bakeries for raising dough. Other enzyme uses

dextrin to make glucose especially during saccarification. As compared to Saccharomyces

cerevisiae, Zymomonas mobilis can also ferment but it is limited to glucose, fructose and sucrose.

(Cardona, 2008).

14
Hydrolysis

Hydrolysis is the process of breaking cellulose and hemicellulose into simple sugar

monomers that can be fermented (Sarkar et al. 2012). There are different hydrolysis strategies

like dilute and concentrated acid, alkaline, hot compressed water, and enzymatic (Yu et al.,

2008). Enzymatic hydrolysis is the most widely used as it is the most eco-friendly. On the other

hand, alkaline and acid hydrolysis present high toxicity, high utility cost, lower sugar yields, and

corrosion, along with the formation of inhibitors (Sarkar et al. 2012).

In chemistry, hydrolysis is a chemical process in which a molecule of water is added to a

substance. Sometimes this addition causes both substance and water molecule to split into two

parts. In such reactions, one fragment of the target molecule (or parent molecule) gains a

hydrogen ion.

Coconut Palm (Cocos nucifera)

Coconut palm (Cocos nucifera) is an important commercial crop in many tropical

countries, but its industry generates large amounts of residue. One way to address this problem is

to use this residue, coconut husk, to produce second-generation (2G) ethanol (Telleria et al.,

2018).

Coconut palms usually have just one upright trunk with non-showy, light tan bark. They

can become up to 60 feet tall and 25 feet wide at the top. Cocos nucifera also comes in smaller

dwarf varieties that grow just 10 to 15 feet tall. Coconut harvesting time is determined by its

purpose and is usually carried out in two stages of ripening. The green fruits are destined to the

coconut water market, while mature fruits are destined to the dry coconut market (for meat, milk

and oil) (Martins et al). The estimated annual worldwide coconut production in 2015 was around

15
55 million tons and the main producing countries are Brazil, India, Indonesia, the Philippines,

and Sri Lanka (FAOSTAT, 2013). However, 80 to 85% of the weight of the fruit is not used and

is simply discarded, resulting in large amount of waste (Montenegro et al., 2001). Possible

solution from the coconut husk residue is the production of 2G bioethanol.

16
METHODOLOGY

Research Design

The researchers will use experimental design in the study. The experimental design is

regarded as the most accurate form of experimental research; it mathematically proves or

disproves a hypothesis with statistical analysis (Antonio, et al 2015). This study will use

experimental research design to determine the effectiveness of the proposed materials as an

alternative main ingredient in making bioethanol and to show the interpretation of data for the

variable being manipulated.

Materials and Procedures

The researchers will adapt a procedure that will be used for the conduction of their

experiment. All the methods were validated and adapted from previous studies and literature.

The coconut shells that will be used will be collected in the the vicinity of Concepcion,

Tarlac. The coconut shells will undergo pretreatment, where the recalcitrant structure of

lignocellulose is disrupted resulting in breakage of lignin sheath, degradation of hemicellulose

and reduction in crystallinity and degree of polymerization of cellulose and alkaline

pretreatment which is a widely studied chemical pretreatment method which is based on the

solubilization of lignin in the alkaline solution. The samples will also undergo dilute acid

hydrolysis in order to break down the cellulose and hemicellulose polymers in lignocellulosic

biomass to form individual sugar molecules which can be fermented into ethanol. (Sidiras and

Koukios, 2004). Fermentation will also be conducted where yeast (Saccharomyces cerevisiae) is

added to the solution, and then fermented for days (5,7 and 9 days) (Oyeleke and Jibrin, 2009).

The last procedure is distillation which is the most dominant and recognized industrial

17
purification technique of ethanol (Oyeleke and Jibrin, 2009). Distillation setup will be used in

order to produce the bioethanol by distillation process that will be conducted in the laboratory.

Locale

The experimentation will be conducted at St. Jude Village, Concepcion, Tarlac and the

school laboratory of Benigno S. Aquino National High School, Senior High School Department,

San Nicolas Poblacion, Concepcion, Tarlac

Materials

Coconuts shells will be collected in the vicinity of Concepcion, Tarlac. Baking yeast

(Saccharomyces cerevisiae) will be used in this study. All chemicals that will be used in this

study are all technical grade and from Dalkem Corporation.

Pretreatment

The pretreatment of the samples will be carried out according to the methods described

by Jannah and Asip, (2015). 1500g of coconut shell will be cleaned until the excess coconut

fiber and coconut will be removed and will be washed with tap water in order to remove any

gross dirt and contamination. The samples will be washed again to ensure complete washing then

the samples will be gradually collected from the top until there was only a small fraction left in

the dirty water. The washed samples will then be sun-dried for 6 hours daily for a period of 1

week to reduce its moisture content and will be crushed and grounded into small pieces using a

mallet.

18
Alkaline Pretreatment

The 1500g of dried grounded coconut shells will be immersed for 3 hours in solution of

Sodium Hydroxide (NaOH) with 35% concentration (Jannah and Asip, 2015). After that, the

coconut shells will be rinsed with distilled water and then sun-dried for 6 hours.

Acid Hydrolysis

100g of each dried pretreated coconut shell samples will be diluted with 200mL aqueous

into a glass beaker, will be adjusted 150mL of Sulfuric acid solution at concentration of 4%

(v/w) and will be heated to a temperature of 90℃ for 60 minutes with continuous stirring to

effect hydrolysis. Reaction of slurry with acid solution will be accelerated by additional heating.

(Jannah and Asip, 2015)

Fermentation

Fermentation of the hydrolyzed samples will be carried out according to the methods

described by Oyeleke and Jibrin, (2009). 50g of yeast (Saccharomyces cerevisiae) will be added

each into the slurry and stirred. Fermentation process will be done with various times (5, 7, and

9 days). The samples will be kept at a temperature of 27℃ which is the standard room

temperature.

Distillation

The slurry will be filtered out using a sieve to separate large particles from the solution

and will be filtered out again using a filter paper. The slurry will then be purified and distilled

19
using the distillation setup that will be conducted at the laboratory to separate the ethanol.

(Oyeleke and Jibrin, 2009)

TESTING

Four sets of samples will be prepared. The three sets will be the distillate of the samples

fermented for 5,7, and 9 days and the fourth set is the ethanol labeled as the commercial one. The

samples will be compared in terms of the volume and density of ethanol produced during the

distillation process to determine any significant difference based on the fermentation time on

ethanol production. (Humphrey and Okafogu, 2007). The samples will also be tested in terms of

their color and combustibility and will be compared to the commercial one. (Pereira, Aleiferis

and Richardson, 2012). The color of the flame, smoke and the duration of the flame between the

fermented samples will be observed.

Determination of Quantity of Bioethanol Produced

The distillates that will be collected will be measured using a measuring cylinder, and

expressed as the quantity of bioethanol produced in g/L by multiplying the volume of the

distillates collected by the density of ethanol (0.8033g/mL) (Humphrey and Okafogu, 2007).

Determination of Density of Bioethanol

The measured volume from each sample will be weighed and divided by the volume

measured.

mass
Density 
volume

20
Combustion Test

The ethanol that will be separated during the distillation will be transferred to a large test

tube, added by a boiling chip, held with a test tube holder and will be heated until the liquid was

boiling. The open end of the test tube will be held to the flame and ignite the ethanol vapors.

Ethanol burns with a pale blue flame with no smoke. (American Herbal Products Association,

2017)

STATISTICAL TREATMENT

The data to be gathered during the experiment will be presented using series of tables

which will indicate any significant difference on the amount of volume produced during the

ethanol production based on the fermentation time from the 3 different fermented samples and to

compare the produced ethanol from the commercial one. To interpret the data effectively, the

researchers will employ the Analysis of Variance (ANOVA).

Analysis of Variance (ANOVA)

This statistical treatment will be used to test the differences between the means of amount

of volume produced during the ethanol production based on the fermentation time from the 3

different fermented samples. ANOVA is used to compare the means between two or more

groups of values (Frontline Solvers, 2019). The analysis will help the researchers in making

comparison of two or more means which will enable them to draw various predictions about the

set of data.

21
BIBLIOGRAPHY

L. Veeranjaneya Reddy and O. Vijaya Sarathi Reddy, (2007) Production of Ethanol from Mango

(Mangifera indica L.) Fruit Juice Fermentation. Research Journal of

Microbiology Volume 2

Mustafa Vohra, Jagdish Manwar, Rahul Manmode, Satish Padgilwarb, Sanjay Patila, (2014).

Bioethanol production: Feedstock and Current Technologies. Journal of Environmental

Chemical Engineering Volume 2

Kevin A Gray et al. (2006). Bioethanol. Retrieved from

https://www.sciencedirect.com/science/article/abs/pii/S1367593106000378

Renewable Fuels Association (2018). Ethanol and Marine Engines. Retrieved from

http://www.redfieldenergy.com/wp-content/uploads/2014/06/ethanol_and_marine.pdf

Bio resource Technology (2017) Science Direct: What is Fermentation. Retrieved from

https://www.sciencedirect.com/topics/earth-and-planetary-sciences/fermentation

Anne Marie Helmenstine, Ph.D. (2019). What is Fermentation Definition and Examples.

Retrieved from https://www.thoughtco.com/what-is-fermentation-608199#ethanol-

fermentation

Theresa Philips (2019). An Explanation to the Process of Hydrolysis. Retrieved from

https://www.thebalance.com/what-is-hydrolysis-375589

Angela Ryczkowski, (2018). Facts on Coconut Palm Tress. Retrieved from

https://homeguides.sfgate.com/coconut-palm-trees-51610.html

Ramkumar Nair, (2017). New method makes bioethanol from waste -- in existing plants.

Retrieved from https://www.sciencedaily.com/releases/2017/10/171030131243.htm

22
Ashok Pandey, (2015). Pretreatment of Biomass. Retrieved from

https://doi.org/10.1016/B978-0-12-800080-9.01001-X

ETIP (2014) Ethanol fermentation. Retrieved From

http://www.etipbioenergy.eu/value-chains/conversiontechnologies/ethanol-fermentation

Jannah and Asip, (2015) Bioethanol Production from Coconut Fiber using Alkaline

Pretreatment and Acid Hydrolysis Method Retrieved from

https://pdfs.semanticscholar.org/4a13/5aef5279f7def8e8854ae1eb123964b2b687.pdf

Pereira, Aleiferis and Richardson, (2012) An Analysis of the Combustion Behavior of Ethanol,

Butanol, Iso-Octane, Gasoline, and Methane in a Direct-Injection Spark-Ignition

Research Engine. Retrieved from.

https://www.nap.edu/read/20284/chapter/8#79

Frontline Solvers, (2019). ANOVA: SINGLE FACTOR. Retrieved From

https://www.solver.com/anova-single-factor

Hossain Zabed, (2013) Bioethanol Production from Fermentable Sugar Juice. Retrieved from.

https://www.hindawi.com/journals/tswj/2014/957102

American Herbal Products Association, (2017) Ethanol Identity Test Methods. Retrieved from.

http://www.ahpa.org/DesktopModules/EasyDNNNews/DocumentDownload.ashx?portali

d=0&moduleid=1244&articleid=875&documentid=369

23