Potentials of Different Fruit Peelings as Substrate for Ethanol Production

A Science Investigatory Project

Presented to the

14th BIOTA Regional Student Research Fair

Naoe, Adrianne Paul

Sapeda, Shiela Mae
Flores, Michelle
Brillo. Mercel
Sare, Arjey
Gantinao, Juan Luis
Alvarez, Sofia Anne

Filamer Christian University

Senior High School Department

February 2018
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Potentials of Different Fruit Peelings as Substrate for Ethanol Production

Abstract

This study seeks to find out the potentials of different fruit peelings for the production of ethanol in terms
of properties such as flammability, volatility, boiling point, and density. The treatments prepared were:
Treatment A- 50 grams Banana (Musa acuminata) Peelings, 150 mL distilled water, 10 grams of Active-
Dry Yeast; Treatment B - 50 grams Mango (Mangifera indica var. carabao) Peelings, 150 mL distilled
water, 10 grams of Active- Dry Yeast; Treatment C - 50 grams Chico (Manilkara zapota) Peelings, 150
mL distilled water, 10 grams of Active- Dry Yeast; and Positive Control (Commercial Ethanol). The raw
materials were gathered and prepared by sun drying and blending. The peelings were then pre-treated
with 2% Hydrogen Peroxide solution for 8 hours. The pre-treated fruit peelings were rinsed three times
and fermented using Active- Dry Yeast for 12 days. Eventually, the treatments were filtered and distilled
until the required amount for experimentation was produced. The produced ethanol was then tested for its
properties in terms of Flammability, Boiling Point, Density, and Volatility. The results of the
flammability test showed that all treatments produced a bluish white flame except for treatment A that
produced a red orange flame. Furthermore, tests for Ethanol properties showed that Treatment C (Chico
Peelings)

substrate for Ethanol Production.

 3

Acknowledgement

The researchers would like to express their gratitude to all the people who inspired them
from the first day until they finished their manuscript. They sincerely express their gratitude to
the following:

First and foremost, the Almighty God for the unconditional guidance, wisdom and
perseverance that He has bestowed to the researchers,

Mr. Jorgen M. Gregorio, High School Principal, for his favorable response and approval
for to conduct this study;

Mrs. Sheila C. Suresca, Senior High School Coordinator, for her warm encouragement,
motivation and trust to the researchers that they can complete this study.

Dr. Erlyn C. Beup, Research Director, for her encouragement, support and for letting the
Researchers use the resources available in the Research Office;

Mr. Leo Art Diosep E. Borres, Research/Capstone Adviser, for his patience and
wholehearted guidance for the improvement of the research paper, enthusiastic encouragement,
help in statistical analysis and useful critiques for this research endeavor;

Mr. Edwin Borja, DENR Officer, for the identification of the fruit peelings;

Ms. Retchel A. Benliro, Class Adviser, for her great moral support and encouragement;

Mr. Fermin John Blanco, Laboratory Custodian, for the approval to use the laboratory;

Paul Christian M. Alado and his group members, for the sharing of ideas as well as
support for the conduct of the study;

To all the friends, classmates and special love ones of the researchers for their undying
love, concern, and support;

And lastly, the researchers own their deepest gratitude to their parents for their
unconditional support, financially, spiritually and emotionally. This accomplishment would not
have been possible without them. Thank you very much.
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Introduction

Background of the Study

Global warming, urban pollution and reserves depletion of fossil fuels have been the
driving forces for current research on the use of alternative energy sources, especially those
derived from biomass. Over the past two decades, the environmental impacts of producing
ethanol have been greatly reduced. Meanwhile, the ecological impacts of petroleum extraction,
refining and use continue to worsen (Renewable Fuels Association, 2018).

Using ethanol as a fuel decreases fossil fuel consumption and increases energy supply
security. It can be used neat or blended with gasoline. Additionally, it is considered
biodegradable and sulphur free. Ethanol usage also resulted to a smaller carbon footprint and an
increase in energy efficiency. Ethanol use reduces greenhouse gas emissions by 43% compared
to gasoline. Ethanol also reduces emissions of particulate matter, carbon monoxide, and volatile
organic compounds, displacing toxic aromatics such as benzene and toluene (Renewable Fuels
Association, 2018).

Nowadays, crops and biomasses are the main sources used for ethanol production.
Ethanol is produced from biomass mostly via a fermentation process using glucose derived from
sugars, starch or cellulose as raw materials (Bioenergy Australia, 2016). To achieve significant
economic and environmental benefit, large amounts of food wastes can be utilized to produce
ethanol. Utilization of discarded materials such as fruit peelings for bioethanol production is one
of the best options.

With the premise presented, the fruit peelings of Banana (Musa acuminata), Chico
(Manilkara zapota) and Mango (Mangifera indica var. carabao) which are widely available in
Capiz but are often discarded contain valuable components such as sucrose, glucose, fructose
and other nutrients that are ideal for fermentation and ethanol production which prompted the
researchers to begin investigating the potentials of different fruit peelings as substrates for
alcohol production.
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Objectives of the Study

Generally, this study seeks to find out the potentials of different fruit peelings as substrate
for ethanol production.

Specifically this study aims to seek out the following:

1. Determine the potentials of different fruit peelings as substrate for as substrate for
ethanol production in terms of flammability, volatility, density and boiling point.

2. Determine if there is a significant difference between the potentials of different fruit

peelings as substrate for ethanol production in terms of flammability, volatility,
density and boiling point.

Hypothesis

Based on the problems presented, the following hypothesis is assumed:

1. There is no significant difference on the potentials of different fruit peelings as

substrate for ethanol production in terms of Flammability, Volatility, Boiling point,
Density,

Significance of the study

Since the fruits peelings used are widely spread throughout the country and are readily
available, the utilization of these peelings would be easy. Economy today is life-lined by
transport, and fuel is the blood that powers our economy.

This study would give a great impact on the fuel crisis that we are facing. The usage of
fruit peelings as substrates for ethanol production is environmentally friendly since peelings are
biodegradable and are considered waste materials.

The amount of disregarded fruit peelings that eventually become pollutants if

accumulated will be decreased and will be put to good use. Converting a renewable non-fossil
carbon, such as organic wastes and biomass consisting of all growing organic matter (plants,
grasses, fruit wastes and algae) to fuel would assure a continua l energy supply (Wyman, 1996).
 6

Scope and Delimitation of the Study

This experimental research was conducted to determine the potentials of different fruit
peelings as substrate for ethanol production. The study is only limited to comparing the
flammability (by determining the color of the flame produced), volatility, boiling point and
density of the produced ethanol from the different treatments.

This study applied the Completely Randomized Design and was conducted at the
Chemistry Laboratory of Filamer Christian University from February 7 to February 21, 2018
under the supervision of Mr. Leo Art Diosep E. Borres, Research Adviser.

Three treatments with three replicates were prepared using the fruit peelings of Banana
(Musa acuminata), Chico (Manilkara zapota) and Mango (Mangifera indica var. carabao) as
substrate for ethanol production. Commercial ethanol was used as the control set-up.

The peelings were pre-treated with 2% Hydrogen Peroxide solution for 8 hours,
fermented using Active- Dry Yeast for 12 days, filtered and distilled until enough amount of
ethanol for experimentation was produced. The products were then tested in terms of
flammability, volatility, boiling point, and density.

Results were then gathered, recorded and analyzed using Mean and One-Way Analysis of
Variance set at 5% alpha level of significance to determine the significant difference between the
potentials of different fruit peelings as substrate for ethanol production in terms of flammability,
volatility, boiling point, and density.
 7

Review of Related Literature

Banana (Musa acuminata)

Banana peel is a rich source of starch (3%), crude protein (6-9%), crude fat (3.8-11%),
total dietary fibre (43.2- 9 % α-
linolenic acid, pectin, essential amino acids (leucine, valine, phenylalanine and threonine) and
micronutrients (K, P, Ca, Mg). Banana peels are also a good source of lignin (6-12%), pectin
(10-21%), cellulose (7.6-9.6%), hemicelluloses (6.4-9.4%) and galactouroninc acid. Banana peel,
an agro waste can be used as a substrate for ethanol production owing to its rich carbohydrate,
crude proteins and reducing sugars. Since banana peels contain lignin in low quantities
(Hammond et al., 1996), it could serve as a good substrate for production of value-added
products like ethanol.

Mango (Mangifera indica var. carabao)

Mango waste is either used as cattle feed or dumped in open areas, where it adds to
environmental pollution. Mango is processed to a maximum extent, thereby pro-ducing high
quality of solid and liquid wastes. Solid wastes, stones, stalks, trimmings and fibrous
materials are obtained during the preparation of raw material. This contributes about 40 to
50% of total fruit waste out of which, 5 to 10% is pulp waste and 15 to 20% is kernel
(Anonymous, 2004; Madhukara et al., 1993; Maini et al., 2000; Pandey et al., 2000). Since
mango is mass produced a lot of waste materials such as peelings can be utilized in ethyl alcohol
production to not only help in the fuel crisis butr to also reduce pollution.

The use of mango peel as a source of pectin and fibre production also has been reported
(Pandia et al., 2004) It also contains high concentrations of sugars which is ideal in ethanol
production.
 8

Chico (Manilkara zapota)

Manilkara zapota, commonly known as Sapodilla, chickoo or sapota, one of the wonders
of nature belongs to family Sapotaceae including about 65 genera and 800 species. The name
Sapodilla is taken from the Spanish word zapotilla which means sapote (a soft edible fruit).
Being a very popular fruit crop, grows well in tropical conditions and cultivated world over in
tropical countries for various benefits like edible fruits, timber, latex,. Sapodilla has its origin in
Mexico and is native to Central America, although it is also cultivated in Asian countries
including India. Sapodilla fruit holds great nutritional value because of good source of sugar
(fructose and sucrose) varying from 12 to 14%. Bose & Mitra reported that a 100 g of edible fruit
contains moisture (73.7 g), carbohydrates (21.49 g), protein (0.7 g), fat (1.1 g), calcium (28 mg),
phosphorus (27 mg), Iron (2 mg) and ascorbic acid (6 mg) (Bano and Ahmed, 2017).

Ethanol

Ethyl Alcohol (or ethanol) is a clear, colorless liquid with a characteristic, agreeable odor.
In dilute aqueous solution, it has a somewhat sweet flavor, but in more concentrated solutions it
has a burning taste.

Ethanol melts at –114.1°C, boils at 78.5°C, and has a density of 0.789 g/mL at 20°C. Its
low freezing point has made it useful as the fluid in thermometers for temperatures below –40°C,
the freezing point of mercury, and for other low-temperature purposes, such as for antifreeze in
automobile radiators (Prof. Shakhashiri, 2009). Ethanol is used as a common solvent in
cosmetics such as perfumes, food colourings and flavourings such as Vanilla essence, medicinal
preparations such as antiseptics, and some cleaning agents. Plants like corn or sugarcane
produced ethanol by utilizing enzymes to convert to simple sugars and yeast to ferment the
sugars.
 9

Methodology

Materials and Tools

This study utilized 50 grams of Mango Peelings, 50 grams of Banana Peelings, 50 grams
of Chico peelings and 30 grams of Active Dry Yeast. The tools used are Erlenmeyer flask,
grammer, digital grammer, beakers, rubber bands, cheese cloth, 4 corks, laboratory thermometer,
watch glass, test tube, iron stand with iron ring, wire gauze, alcohol lamp, glass tubing,
condenser, distilling flask, multi clamp, and universal clamp.

Research Design

This study utilized the Completely Randomized Design. In a completely randomized

design, treatment levels or combinations are assigned to experimental units at random. This is
typically done by listing the treatment levels or treatment combinations and assigning a random
number to each (PennState, 2010).

Experimental Layout

Table 1. Different Treatments and Replicates

Replicates
Treatments R1 R2 R3
A- 50 grams Banana (Musa acuminata) Peelings,150 mL
distilled water, 10 grams of Active- Dry Yeast;
B - 50 grams Mango (Mangifera indica) Peelings, 150
mL distilled water, 10 grams of Active – Dry Yeast;

C – 50 grams Chico (Manilkara zapota) Peelings , 150

mL distilled water, 10 grams of Active – Dry Yeast;
Positive Control –
Commercial Ethanol
 10

Procedural Design

Gathering and Preparing of Materials and Tools

Removing the Peelings from the Fruit Weighing 10 g of Active-Dry Yeast

Drying the Fruit Peelings

Blending the Fruit Peelings

Weighing the Fruit Peelings

Alkaline Pretreatment of Peelings

Rinsing the Treated Fruit Peelings

Placing the Fruit Peelings in Beakers

Adding the Water and Yeast to the Fruit Peelings

for Fermentation

A- 50 grams Banana B - 50 grams Mango C – 50 grams Chico

(Musa acuminata) (Mangifera indica) (Manilkara zapota)
Peelings,150 mL Peelings, 150 mL Peelings, 150 mL
distilled water, 10 grams distilled water, 10 grams distilled water, 10 grams
of Active- Dry Yeast; of Active – Dry Yeast; of Active – Dry Yeast;

Filtering the Fermented Treatments

Distilling the Fermented Treatments

Testing the Collected Ethanol from the Treatments

Flammability Flammability Flammability Flammability

 11

General Procedure

Gathering and Preparation of Materials

The fruits were gathered from the local fruit market. Slightly rotten and discolored fruits
were selected. Then the fruits were washed thoroughly as well as the laboratory tools used. The
fruits were then peeled and the peelings were sun dried. After dying, the fruits were weighed,
blended and placed on beakers according to the treatments specified.

Preparation of Materials and Tools

Photo by: Sofia Anne Alvarez

Alkaline Pre-treatment

The prepared fruit peelings were pretreated with 2% Hydrogen Peroxide (H2O2)
solution. The fruit peelings were soaked for 8 hours at room temperature. After 8 hours the
solutions were drained and the peelings were rinsed three times using distilled water. The residue
was then placed again inside the cleaned beakers. The purpose of the alkaline pretreatment was
delignification. The removal of lignin is necessary for cellulose to become readily available for
the enzymes, which permit the yeast to convert the glucose into ethanol (Wyman, 1996).

Alkaline Pretreatment
Photo by: Mercel Brillo
 12

Fermentation of Fruit Peelings

After the pre-treatment, 10 grams of active-dry yeast and 150 mL distilled water was
added to each set-up and stirred until thoroughly mixed. The treatments were then covered with
cheese cloth and left to ferment for 12 days at room temperature.

Distillation

After 12 days, the fermented fruit peelings were filtered and placed on a distilling flask
covered by a rubber stopper with the laboratory thermometer on its hole. The distilling flask was
arranged on an iron stand and connected with the condenser for efficient distillation, with
another flask at the end of the condenser to receive the distillate. The set-up was heated to 78 °C,
the boiling point of ethanol.

Fermentation of Treatments Distillation of Treatments

Photo by: Shiela Mae Sapeda Photo by: Adrianne Paul Naoe

Testing the Product

After collecting enough amount of ethanol for experimentation, the following tests were
conducted:

Volatility. 1 mL of bioethanol produced was placed in a watch glass. The time it takes to
evaporate was then recorded.

Boiling Point. 1 mL of the bioethanol produced was placed in a test tube. The
temperature of its boiling point was then recorded.

Density. 1 mL of mixture from each treatment was used to obtain its density. Each was
weighed in an electronic balance. The data was measured using the formula: Density=
Mass/Volume

Flammability. 1 mL bioethanol produced was placed in a watch glass. The color of the
flame was then recorded.
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Volatility Test Test for Boiling Point

Photo by: Paul Christian Alado Photo by: Alaiza Kaye Dordas

Flammability Test Density Test

Photo by: Paul Christian Alado Photo by: Alaiza Kaye Dordas

Data Analysis

The data were gathered, recorded and tabulated. The potentials of different fruit peelings
as substrate for ethanol production were determined by comparing the results to the standard
properties of commercial ethanol in terms of volatility, boiling point, density and flammability.
After which, the results were ranked for each treatment. The total rank (accumulated rank for
each property) with the least number was computed to determine the treatment with the greatest
potential to be used as substrate for ethanol production (Frenila, 2011).

Mean and One Way Analysis of Variance (ANOVA) set at 5% alpha level of significance
was used in analyzing the results and determining the significant differences between each
treatment.
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Results and Discussion

Table 2. Results for Volatility Test (in seconds)

Replicates
Treatment 1 2 3 Mean
A- 50 grams Banana Peelings,150
mL distilled water, 10 grams of 55 56 56.4 55.80
Active- Dry Yeast;
B - 50 grams Mango Peelings, 150
mL distilled water, 10 grams of
48 47 45 46.67
Active – Dry Yeast;
C – 50 grams Chico Peelings , 150
mL distilled water, 10 grams of
45.6 46 45.8 45.80
Active – Dry Yeast;
Positive Control –
Commercial Ethanol
43 44 42 43.00

Data for Table 2 shows that Treatment C obtained the closest time it takes for ethanol to
evaporate as compared to commercial ethanol with 45.80 seconds, followed by Treatment B with
46.67 seconds and lastly, Treatment A with 55.80 seconds. The positive control gained an
average time of 43.00 seconds.

Table 3. One-way ANOVA for Volatility Test

Source SS df MS F p-value
Treatment 276.970 3 92.3233 94.85 1.36E-06
Error 7.787 8 0.9733
Total 284.757 11

One Way Analysis of Variance for Volatility Test showed that p-value (1.36E-06) is less
than the alpha level of significance (0.05), thus rejecting the null hypothesis. Therefore, there is a
significant difference on the potentials of different fruit peelings as substrate for ethanol in terms
of volatility.
 15

Table 4. Results for Boiling Point (in °C)

Replicates
Treatments 1 2 3 Mean
A- 50 grams Banana Peelings,150
mL distilled water, 10 grams of 90 80 80 83.3
Active- Dry Yeast;
B - 50 grams Mango Peelings, 150
mL distilled water, 10 grams of 78 77 77 77.3
Active – Dry Yeast;
C – 50 grams Chico Peelings, 150
mL distilled water, 10 grams of 78 78 77 77.7
Active – Dry Yeast;
Positive Control –
Commercial Ethanol 78 78 78 78.0

Data for Table 4 shows that Treatment C obtained the closest boiling point temperature as
compared to commercial ethanol with 77.7 °C, followed by Treatment B with 77.3 °C and lastly,
Treatment A with 83.3 °C. The positive control gained a boiling point temperature of 78 °C.

Table 5. One-way ANOVA for Boiling Point Test

Source SS df MS F p-value
Treatment 72.92 3 24.306 2.86 .1044
Error 68.00 8 8.500
Total 140.92 11

One Way Analysis of Variance for Boiling Point Test showed that p-value (.1044) is
greating than the alpha level of significance (0.05), thus accepting the null hypothesis. Therefore,
there is no significant difference on the potentials of different fruit peelings as substrate for
ethanol in terms of boiling point.
 16

Table 6. Results for Density (in g/mL)

Replicates
Treatment 1 2 3 Mean
A- 50 grams Banana Peelings,150
mL distilled water, 10 grams of 0.75 0.74 0.76 0.750
Active- Dry Yeast;
B - 50 grams Mango Peelings, 150
mL distilled water, 10 grams of 0.69 0.68 0.69 0.687
Active – Dry Yeast;
C – 50 grams Chico Peelings, 150
mL distilled water, 10 grams of 0.75 0.76 0.77 0.760
Active – Dry Yeast;
Positive Control –
Commercial Ethanol 0.80 0.79 0.90 0.830

Data for Table 6 shows that Treatment C obtained the closest density as compared to
commercial ethanol with 0.760 g/mL, followed by Treatment A with 0.750 g/mL, and lastly,
Treatment B with 0.687 g/mL. The positive control gained a density of 0.830 g/mL.

Table 7. One-way ANOVA for Density Test

Source SS df MS F p-value
Treatment 0.0310 3 0.01033 10.51 .0038
Error 0.0079 8 0.00098
Total 0.0389 11

One Way Analysis of Variance for Density Test showed that p-value (.0038) is less than
the alpha level of significance (0.05), thus rejecting the null hypothesis. Therefore, there is a
significant difference on the potentials of different fruit peelings as substrate for ethanol in terms
of density.
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Table 8. Results for Flammability Test (Color of Flame)

Replicates
Treatments 1 2 3 Dominant
Color
A- 50 grams Banana Peelings,150
mL distilled water, 10 grams of Red-orange Red-orange Red-orange Red-orange
Active- Dry Yeast;
B - 50 grams Mango Peelings, 150
mL distilled water, 10 grams of Bluish-white Bluish-white Bluish-white Bluish-white
Active – Dry Yeast;
C – 50 grams Chico Peelings , 150
mL distilled water, 10 grams of Bluish-white Bluish-white Bluish-white Bluish-white
Active – Dry Yeast;
Positive Control –
Commercial Ethanol Bluish-white Bluish-white Bluish-white Bluish-white

Data for Table 8 shows that Treatment B, C and Positive Control has the same flame
color which is bluish-white whereas Treatment A produced a red-orange flame. The bluish flame
is an indication that it has a higher temperature compared with other flame colors (Frenila,
2011).

Table 9. Ranking of the Different Fruit Peelings (as compared to Commercial Ethanol) in
Terms of their Potential as Substrate for Ethanol Production in Terms of Volatility, Boiling
Point, Density and Flammability
Rank Rank
Treatment Volatility Boiling Density Flamm Rank in in Rank in Total Substrate
(s) Point (g/mL) -ability Volatility Boiling in Flamm Rank Potential
(°C) Point Density -
ability
A Red-
(Banana 55.80 83.3 0.750 3 3 2 3 11 3
orange
Peelings)
B Bluish
(Mango 77.3 0.687 -White
Peelings) 46.67 2 2 3 1.5 8.5 2
C Bluish
(Chico 77.7 0.760 -White
Peelings) 45.80 1 1 1 1.5 4.5 1
 18

Table 9 shows the results of the ranking of the different fruit peelings as potential
substrate for ethanol production considering the volatility, boiling point, density and
flammability. It is shown in the table that Treatment C (Chico Peelings) ranked first as the fruit
peelings with the greatest potential to be used as substrate for ethanol production with a total
rank of 4.5, followed by Treatment B (Mango Peelings) with a total rank of 8.5, and lastly,
Treatment A (Banana Peelings) with a total rank of 11.

Discussion
Based on the aforementioned results, Treatment C (Chico Peelings) displayed the greatest
potential as substrate for ethanol production as it ranked first, followed by Treatment B (Mango
Peelings) and lastly, Treatment A (Banana Peelings). Throughout the study, the results
unanimously showed that treatment C (Chico) was the closest to commercial ethanol with a
mean of 45.80 seconds in terms of volatility, 77.7 °C in terms of Boiling point, 0.760 g/mL in
terms of density, and a bluish-white flame color.
Furthermore, One-way Analysis of Variance shows that there is a significant difference
on the potentials of different fruit peelings when used as substrate for ethanol production in
terms of volatility (1.36E-06 < 0.05) and density (0.0038 < 0.05). However, statistical analysis
also showed that there is no significant difference when it comes to boiling point (0.1044 >
0.05). The significant difference of the results signifies that the set-up was controlled and the
results did not occur by coincidence.
The results were consistent on the findings of Bano and Ahmed (2017) that chemical
properties of Chico is composed of varying amounts of sugar (fructose and sucrose) from 12 to
14%. Lastly the results strengthen the concept that fruit peelings contain a considerable amount
of fermentable sugars and is an ideal substrate for ethanol production
 19

Conclusion and Recommendations

Conclusions
Based on the aforementioned findings, the following conclusions were drawn:
1. Chico peelings has the greatest potential to be used as substrate for ethanol production in
terms of volatility, boiling point, density and flammability, followed by mango peelings
and lastly, banana peelings.
2. There is a significant difference on the potentials of different fruit peelings as substrate
for ethanol production in terms of volatility and density. However, there is no significant
difference in terms of boiling point.

Recommendations
Based on the conclusions drawn, the researchers strongly recommend the following:
1. Promote the utilization of discarded fruit peelings, especially of Chico, as substrate to
produce ethanol since it was found out that the ethanol produced from Chico and other
fruit peelings used in this study is comparable to commercial ethanol.
2. Further studies and experimentation on the usability and application of ethanol produced
from fruit peelings as well as optimization of the process.
3. Further experimentation on other organic wastes or widely available yet discarded
materials as source or ethanol.
 20

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