1 CHAPTER I

2 INTRODUCTION

3 Bananas originated in the Indo-Malaysian region and are grown in every tropical area in

4 the world. Because of the favorable climate, the Philippines produces about 3.5 metric tons of

5 bananas yearly. Banana is also considered as the most important fruit crop in the country in

6 terms of volume of production and export earnings which results to a lot of banana wastes that

7 can eventually cause pollution and other environmental problems. About 382,491 hectares and

8 half of the Philippines' land area are planted with bananas and because of their popularity in the

9 Philippines, it is undeniable that there is a huge amount of banana waste in the country

10 (PBWorks, 2008).

11 The Philippines is the second largest exporter of bananas after Ecuador, with some 2.6

12 metric tons exported in 2012. That year, the exports from the Philippines (essentially Cavendish

13 cultivars) made up 98% of the Asian banana trade. Two-thirds of the exported volumes were

14 shipped to Japan, China, and South Korea. In 2015, the country produced nearly 9.1 metric tons

15 of bananas (Vézina, 2016).

16 This research aims to reduce the amount of banana waste in the Philippines and to know

17 if alcohol can be produced (and its concentration) from banana peelings through the process of

18 fermentation. Fermentation is an anaerobic process that takes place in the absence of any

19 external electron acceptor (Encyclopaedia Britannica, n.d.). Fermentation usually produces lactic

20 acid, hydrogen and ethanol. This process often uses sugar as its common substrate. The

21 hydrolysis of starch to maltose comes first during fermentation with the help of the enzyme

22 called diastase. This enzyme can be obtained by propagating malt or barley seeds. Next, the

23 enzyme maltase converts maltose to glucose. Zymase, another enzyme, then ferments glucose to

1
 1 ethanol. Through this enzyme, one can be able to save money by converting banana peels to

2 produce multipurpose natural alcohol while minimizing the amount of banana peel waste in the

3 process.

4 A few studies have been found that resemble the production of alcohol from bananas.

5 Lakshmi (2017) conducted a research in which waste banana plant parts (leaf, stem) were

6 enzymatically treated with cellulase isolated from fungal stains. He suggested that waste banana

7 plantation destroyed after the banana yield is obtained can be used for the production of ethanol.

8 This study used gas chromatography and chromatic acid test to analyze the ethanol concentration

9 of the sample.

10 On the other hand, Chongkhong (2016) aimed to produce ethanol from banana peels by

11 using a two-step response surface methodology. First, the banana peels were subjected to vinegar

12 hydrolysis using microwave heating and the second process was using commercial baker’s yeast

13 to ferment the peel hydrolysate. This study concluded that the production of bio-ethanol from

14 banana peels by such two-step process appeared technically feasible, and might provide

15 economic advantages. Gas chromatography was used to determine the ethanol content in the

16 fermentation broth.

17 The study by Jagessar and Fraser (2015) used the fermentation of fruit mash of banana,

18 pineapple, and mango to produce ethanol. In over three days, fermentation was done in

19 triplicates and each fruit was subjected to the same experimental conditions. Fehling’s solution

20 was used to test sugar presence after filtering the mixture at the specified time. After the

21 distillation of the filtrate, a Pictometer was used to test alcohol strength. This study concluded

22 that the pulp of bananas, pineapples, and mangoes could be used to produce ethanol for

23 commercial and industrial applications.

2
 1 Manikandan et al. (2007) studied ethanol production employing five mutant strains of

2 Saccharomyces cerevisiae using banana peel as a substrate. The effect of temperature, pH and

3 initial substrate concentration on ethanol production was studied. The ethanol concentrations of

4 the samples in this study were measured using a gas chromatograph and flame ionization

5 detector. It also concluded that ethanol could be produced efficiently by controlled fermentation

6 techniques from agro-residues particularly banana peel waste using mutant strains of

7 Saccharomyces cerevisiae.

8 The study by Ali et al. (2014) also aimed to produce ethanol from plant wastes such as

9 banana peels, which are abundant and do not interfere with food safety using co-culture of

10 baker’s yeast or Saccharomyces cerevisiae. In this study, the instrument called Dujardin-Salleron

11 Ebulliometer was used to quantify the ethanol content. The difference in boiling points between

12 the solution and pure water is integral in finding the ethanol content using the Ebulliometer. The

13 percentage of ethanol can be found based on a comparison using a calculating dial. This study

14 concluded that ethanol could be produced efficiently by controlled fermentation technique from

15 banana peel waste using Saccharomyces cerevisiae.

16 Gebregergs et al. (2016) made industrial ethanol from banana peels for developing

17 countries by using surface response methodology where they dried and ground banana peels

18 before proceeding to the fermentation process which lasted for three days. They used the

19 pycnometer method to determine alcohol concentration. This study also discussed the effects of

20 acid concentration, time, temperature, and the benzathine penicillin to the concentration of

21 alcohol.

22

3
 1 Theoretical Framework

2 Sugar composition in bananas

3 Bananas are an abundant source of carbohydrates. Unripe bananas contain starch and ripe

4 bananas contain sugars. On a dry weight basis, green bananas contain up to 70-80% of starch.

5 During ripening, the carbohydrate composition of bananas changes considerably (Arnarson,

6 2014).

7 The starch is converted into sugars during ripening. When the banana is fully ripe, its

8 starch content ends up being less than 1%. Sucrose, fructose, and glucose are the most common

9 kinds of sugar found in ripe bananas.

10 In this study, starch from bananas were used in the fermentation process which was

11 instrumental to the production of alcohol.

12 Ethanol production & fermentation

13 Alcohol which is also known as ethanol, is a colorless, combustible, hazardous liquid

14 with a molecular formula of C2H6O. It has wide applications in the industry, including its uses as

15 fuel, solvent, beverage, antiseptic, and raw material for the production of other useful chemicals.

16 It is produced from fermentation of starch, sugars, hydrocarbon-based ethanol production, or

17 lignocellulosic materials. One efficient and cost-effective alternative that is still under research is

18 the use of lignocellulosic biomass for bioethanol production. This biomass inludes agricultural

19 remains or municipal and industrial wastes. Sawdust, wood, wheat, sugarcane, bagasse,

20 newspaper, corn curb, and rice straw are some substances that have been used to produce

21 bioethanol (Danmaliki et al., 2016).

22

4
 1 Alcoholic fermentation is an older biochemical pathway which makes use of a crew of

2 bacteria and yeast to make ethanol. Sugars like glucose, sucrose, and fructose are converted into

3 small amounts of ATP to produce ethanol and carbon dioxide during fermentation (ck12.org,

4 n.d.).

5 In the study of Shang-Tian & Yali (2007), many microorganisms, including bacteria and

6 yeasts, can produce ethanol as the major fermentation product from carbohydrates. Current

7 industrial ethanol fermentation is mainly carried out with the yeast Saccharomyces cerevisiae

8 because of its hardiness (low pH and high ethanol tolerance), although the bacterium Zymomonas

9 mobilis has a higher specific ethanol productivity and yield from glucose and sucrose.

10 In this modern era, attention has been paid to the production of ethanol for chemical and

11 fuel purposes by microbial fermentation. Ethanol is nowadays produced by using sugar beet,

12 potatoes, corn, cassava, and sugar cane. Both yeasts (Saccharomyces cerevisiae, S. uvarum S.

13 carlsbergensis, Candida brassicae, C. utilis, Kluyveromyces fragilis, K. lactis) and bacteria

14 (Zymomonas mobilis) have been employed for ethanol production in industries

15 (biologydiscussion.com, n.d.).

16 In this study, yeast will be used. Specifically, baker’s yeast or Saccharomyces cerevisiae

17 will be significant in the fermentation process. Although bacteria can produce ethanol, yeast is

18 more accessible to the researchers which makes it a more desirable and convenient organism to

19 use in this research.

20 In chemical terms, alcoholic fermentation is made up of two phases: first, the sugar is

21 converted into pyruvate and the second phase concerns the conversion of pyruvate into alcohol.

22 Simply, the fermenting step of the chemical reaction is the second phase (Khurana et al., 2017).

5
 1 Nelson & Cox (2008) explained the process of glycolysis in their study. The first major

2 step in fermentation is glycolysis which is the metabolic pathway that converts glucose to

3 pyruvate. This metabolic pathway is considered as ancient because it has probably been

4 developed when no oxygen was present in the environment 3.5 billion years ago. Glycolysis not

5 only occurs in microorganisms like bacteria because it happens in every living cell.

6 In ethanol fermentation, two important steps are needed to convert the pyruvate produced

7 through glycolysis to ethanol and carbon dioxide. First, acetaldehyde, a two-carbon compound, is

8 formed because pyruvate released carbon dioxide. Next, nicotinamide adenine dinucleotide

9 (NADH) reduces acetaldehyde to ethanol which regenerates the NAD+ for use in glycolysis. As

10 a whole, one molecule of glucose is converted into two molecules of ethanol and two molecules

11 of carbon dioxide (biologydictionary.net, n.d.).

12 In anoxygenic conditions (absence of oxygen), pyruvic acid can follow two different

13 paths, depending on the cell type. Through the the alcoholic fermentation pathway, it can be

14 converted into ethanol (alcohol) or it can be converted into lactate through the lactic acid

15 fermentation pathway (Alba-Lois & Segal-Kischinevzky, 2010).

16

6
 1 Conceptual Framework

2 Input Process Output

3
 Gather the banana
peelings.
4  Dry the banana
peelings under the sun.
Put them in the oven to
5 dry further if needed.
 Grind the dried banana
 Banana peelings peelings.
6 The concentration of
 Three containers  The fermentation
 Knife process will follow. ethyl alcohol from
 Water  Filter unnecesary banana peelings
7
 Yeast particles before
distillation.
8  Get alcohol through
distillation.
 Investigate alcohol
9 yield and determine the
alcohol presence,
volume, and
10 concentration.
 Analyze data.
11

12 Figure 1. The Research Conceptual Framework

13
14
15
16 Statement of the Problem

17 The researchers would like to find out if ethyl alcohol can be produced from banana

18 peelings. Specifically, this research aims to answer these following questions:

19 1. Can alcohol be produced from banana peelings of:

20 a.) Musa sapientum (latundan)

7
 1 b.) Musa acuminata x balbisiana (saba)

2
3
4
5 Research Hypotheses

6 Null hypothesis: Alcohol cannot be produced from the banana peelings of Musa

7 sapientum (latundan) and Musa acuminata x balbisiana (saba).

8 Alternative hypothesis: Alcohol can be produced from the banana peelings of Musa

9 sapientum (latundan) and Musa acuminata x balbisiana (saba).

10
11
12
13 Significance of the Study

14 This study is significant because it can help in managing waste through using banana

15 peels as a main material for the production of alcohol. Therefore, this study will be significantly

16 beneficial to the following:

17  Community: This can be beneficial to the community for it can lessen the amount of

18 garbage from banana peels which can bring about pollution.

19  Clinics and hospitals: It can offer a cheaper alternative to the commercial ethyl alcohol

20 used by clinics and hospitals for the sanitation of medical instruments.

21  Future Researchers: This study can shed light on the other uses of banana peels which

22 will be of use to future researchers.

23

8
 1 Scope and Delimitation

2 This study aims to know if alcohol can be produced from banana peelings of Musa

3 sapientum (latundan) and Musa acuminata x balbisiana (saba). It also intends to know the

4 concentration and the volume of alcohol produced from the banana peels of the aforementioned

5 banana varieties. To achieve the desired results, laboratory tests were necessary. Distillation was

6 done to get the alcohol after fermentation. These laboratory tests were conducted inside the

7 Palompon Institute of Technology Chemistry Laboratory.

8 It needs to be noted that only first distillation of the samples was made because the

9 laboratory equipment only allowed a small volume of the samples to be distilled which resulted

10 in a smaller yield. Also, because there was no available pycnometer in the chemistry laboratory,

11 the density of the distillate was determined using a graduated cylinder and an electronic

12 weighing scale.

13 This study was conducted within the first semester of S.Y. 2018-2019.

14
15
16
17 Definition of Terms

18 Alcohol. This is the end goal of the researchers, the proposed product to be made using

19 banana peel as the main ingredient.

20 Amount of alcohol. This refers to the amount of alcohol obtained from banana peelings

21 and the process of fermentation.

9
 1 Banana peelings. This is the outer cover of the banana fruit. It is the main ingredient of

2 the study and will be used to produce alcohol.

3 Concentration. The percentage of alcohol present in the alcohol sample made from

4 banana peelings.

5 Distillation. This is a process of boiling and condensing liquid. This will be done in this

6 study to obtain alcohol after fermentation.

7 Fermentation. This process is the breakdown of carbohydrates (e.g. starch) by

8 microorganisms such as yeast. In this study, fermentation will be done to produce alcohol from

9 banana peels.

10 Starch. It is a carbohydrate substance that is present in banana peels. It is significant in

11 this study because starch is one of the necessary ingredients for fermentation.

10
 1 CHAPTER II

2 METHODOLOGY

3 This chapter presents aspects of the study considered very important, such as: research

4 design, locale of the study, research instruments and experiment procedure.

5 Research design

6 This study adopted a tru experimental research design to produce ethyl alcohol from

7 banana peelings. It aims to determine its concentration and the amount of alcohol produced.

8
9
10
11 Locale of the Study

12 Laboratory experiments were necessary to make this study possible. Experiments were

13 done in Palompon Institute of Technology. The researchers made use of the school’s chemistry

14 laboratory and its equipment to be able to achieve the desired results and produce alcohol from

15 peelings of Musa sapientum (latundan) and Musa acuminata x balbisiana (saba).

16
17
18
19 Research Instruments

20 One important material used in this study were the banana peelings. The banana peelings

21 used in this study were of two varieties with the same level of ripeness: Musa sapientum

22 (latundan) and Musa acuminata x balbisiana (saba). These peels were used within 24 hours of

23 collection to utilize the peels before they became overripe and deteriorated. Other important

11
 1 materials used were containers where the samples were stored, distilled water which was

2 essential in fermentation, and baker’s yeast (Saccharomyces cerevisiae) which was also

3 important in the fermentation process. Laboratory equipment such as a 250 mL Erlenmeyer

4 flask, hot plate, condenser, 50 mL beaker, thistle tube, alcohol lamp, rubber stopper with holes,

5 rubber tube, test tube, iron ring, iron stand, wire gauze, and small glass tubes were also used in

6 the simple distillation process.

7
8
9
10

Distilled water
11

12 Banana peelings

13

14

15

16
Laboratory equipment used

17 Container Laboratory equipment used

18

19

20

21 Dried banana peelings

Baker’s yeast
Laboratory equipment used 12
 1 Experiment Procedure

2 1. Preparation of banana peelings

3 The first step in producing ethyl alcohol from banana peels was to get starch from the

4 peels of the two varieties of banana. The peels were of the same mass and underwent the same

5 process. The peelings were sundried for three days and were dried further inside an oven for 15

6 minutes at 60°C. Next, the researchers ground the dried banana peelings until they became

7 pulverized.

8
9
10
11
12
13
14 2. Fermentation

15 The next step was the fermentation process. Three experimental samples were prepared

16 for every banana variety. The samples had the same mass of starch (100 grams), the same mass

17 of yeast (10 grams), and the same volume of distilled water (1 liter). Baker’s yeast,

18 Saccharomyces cerevisiae, was used as the microorganism for alcohol fermentation. It is

19 important to note that fermentation of the samples was done in different durations (e.g., one

20 week, two weeks, one month) to determine the effect of fermentation length on the alcohol

21 concentration. The samples were closely observed and examined during fermentation. The

22 researchers also filtered out unnecessary particles before proceeding to the distillation process.

23

24

13
 1 3. Distillation

2 After fermentation, the alcohol yield from the banana peelings was investigated. The

3 alcohol product from the banana peels was yielded by a simple distillation method. In this

4 process, alcohol was separated from water and other substances. Only first distillation of the

5 samples was made due to the low distillate yield.

10 4. Determining alcohol concentration

11 Alcohol concentration was determined using a graduated cylinder and an electronic

12 weighing scale. After distillation, the mass and volume of what was yielded were recorded. The

13 density of the yield was then computed and compared with the table showing the concentration

14 of ethanol-water mixture in relation to specific gravity at various temperature (see Appendix A).

15 The temperature was also taken into consideration in determining the alcohol concentration.

16

14
 1 CHAPTER III

2 RESULTS AND DISCUSSION

3 This chapter gives a discussion of the study’s results. It also sheds light on the data

4 gathered from the experiment done by the researchers and their corresponding interpretation. The

5 effects of fermentation length, temperature, and other factors to the alcohol presence and

6 concentration are discussed in this chapter as well.

7 General Discussion

8 Throughout this study, the researchers fermented the peelings of two banana varieties—

9 the Musa acuminata x balbisiana (saba) and the Musa sapientum (latundan)—in triplicates. First,

10 the researchers fermented three samples of the Musa acuminata x balbisiana (saba) peelings.

11 Simple distillation of one of the samples came next after one week. The mass and the volume of

12 the yield were noted, and then the researchers computed the density of the yield to determine the

13 alcohol concentration. The density was known to be approximately 0.86 g/cm3. Referring to the

14 table showing the concentration of ethanol-water mixture in relation to specific gravity at various

15 temperature (see Appendix A), the yield turned out to have 70% ethanol concentration. However,

16 the researchers failed to perform the distillation process on the second and third samples of the

17 Musa acuminata x balbisiana (saba) because, by the second week, they emitted a very foul odor.

18 Both samples became spoiled and were discarded.

19 Next, the researchers fermented the dried peelings of the Musa sapientum (latundan).

20 They made three samples for this with each sample fermented for two weeks, three weeks, and a

21 month, respectively. They performed the simple distillation process on one sample first which

22 they fermented for two weeks and determined the density of the yield. This time, the density was

15
 1 approximately 0.91 g/cm3, and when compared to the table, the ethanol concentration was

2 determined to be 48% at 30 degrees temperature. However, when the yield cooled down, the

3 researchers determined its density again and found it increased to 0.96 g/cm3. When it cooled

4 down further, the density increased to more than 1.0 g/cm3 which indicated that temperature

5 affected the concentration of the yield. Other factors that affected the presence of alcohol in the

6 yield will be further discussed below.

7 The second sample of the Musa sapientum (latundan) was distilled after three weeks of

8 fermentation. The same process of determining the density of the yield was done, and it was

9 determined that it exceeded 1 g/cm3 which meant that no alcohol was present in the yield. The

10 same thing happened to the third sample of the Musa sapientum (latundan) when it was distilled

11 after four weeks. The density was determined to be more than 1 g/cm3 as well.

12 It is important to note that the researchers performed the distillation process on various

13 samples over and over, but most of the distillates had a density of 1 g/cm3 or more which

14 indicated that they didn’t yield any alcohol.

15

16
 1 Table 1. Alcohol concentration based on banana variety

Alcohol Concentration
Banana Variety Density

Musa acuminata x balbisiana (saba)

 Sample 1 ~0.86 g/ml 70%

Musa sapientum (latundan)

 Sample 1 ~0.91 g/mL 48%

 Sample 2 ~1.0 g/mL 0

 Sample 3 ~1.2 g/mL 0

2
3
4
5 Effects of fermentation length to alcohol presence and concentration

6 Based on the data above, the researchers found out that fermentation length can affect the

7 presence and concentration of alcohol. Similar to the findings of this study, Hashem et al. (2013)

8 observed the effect of varying fermentation periods on the efficiency in ethanol production of

9 very high ethanol producers and thermotolerant strains, and revealed that the ethanol yield

10 gradually increased with time but recorded optimal results between 60 and 72 hours with a

11 dramatically decreased ethanol production with further extension of fermentation periods. The

12 Musa acuminata x balbisiana (saba) peelings that were distilled for a week yielded the highest

13 alcohol concentration while the Musa sapientum (latundan) that were fermented for a longer

14 amount of time yielded the least alcohol concentration, and the last two samples did not yield

15 any alcohol at all. One factor that could have affected this is the conversion of alcohol to acetic

16 acid. When fermentation is done for too long, the sample can lose its ethanol content because of

17
 1 the growth of acetic acid bacteria. According to ScienceDirect (2011), the completion of

2 alcoholic fermentation may lead to oxygen enrichment which increases the likelihood that acetic

3 acid bacteria will grow because it may activate the metabolism of acetic acid bacteria. It is better

4 to distill the samples before the aforementioned bacteria grows. The data gathered from this

5 study indicate that a week of fermentation is the optimum fermentation length for the production

6 of ethanol.

7
8
9
10 Effects of temperature to alcohol presence and concentration

11 Temperature also affects alcohol presence and concentration. Togarepi et al. (2010)

12 found out that at a temperature of 30 ºC, the maximum rate of ethanol (alcohol) production was

13 achieved. From the experiments done by the researchers, they have learned that alcohol

14 concentration is greater when the temperature of the yield is higher. This is what happened to the

15 second sample of the fermented peelings of the Musa sapientum (latundan) when the alcohol was

16 yielded through distillation. When the yield cooled down, the density changed and increased. It

17 can be said that alcohol concentration is directly proportional to the temperature and inversely

18 proportional to the density. When the temperature of the yield is higher, the concentration is also

19 higher. When the density is lower, the alcohol concentration is greater.

20 Objects expand and become larger when temperature increases and therefore the density

21 decreases. When the temperature decreases, objects condense and become smaller so density

22 increases (sciencing.com, 2018).

23

18
 1 Other factors affecting alcohol presence and concentration

2 There are also more factors that can affect alcohol presence and concentration. One of

3 these is contamination. This may happen because of the constant transfer of the yield from one

4 container to another. The transfer may have lessened the alcohol present in the yield. Another

5 factor that could have affected alcohol presence and concentration is the evaporation of ethanol.

6 Ethanol vaporizes easily so some of the alcohol content from the yield may have evaporated

7 even before its density and concentration were determined (Petesch, 2015). Compared to water,

8 ethanol boils at a lower temperature, which generally means that it will evaporate quicker than

9 water. This is largely determined by attractive interactions between the liquid molecules.

10 Lack of enough equipment may also be one factor that can affect the production of

11 alcohol. In this study, the researchers mostly improvised laboratory equipment to make

12 distillation possible, and this may have had a hand in the contamination of the alcohol yield. The

13 variety and ripeness of banana is also a factor in this study. Musa acuminata x balbisiana yielded

14 a higher concentration of alcohol than Musa sapientum. According to Hammond et al. (1996),

15 the yield per unit weight of the original green bananas tends to be higher when the bananas were

16 fermented green. The yield of the overripe bananas is about 23% lower than that of green

17 bananas.

19
 1 CHAPTER IV

2 CONCLUSION AND RECOMMENDATIONS

3 In this chapter, conclusions are drawn from the data. Recommendations are also made for

4 future researchers and for other people who seek information about the production of alcohol

5 from banana peelings.

6 Conclusion

7 In conclusion, peelings from the banana varieties Musa acuminata x balbisiana (saba)

8 and Musa sapientum (latundan) are suitable to produce alcohol. The experimental results show

9 that alcohol concentration varies due to banana variety and is affected by factors such as

10 fermentation length, temperature, and contamination. The longer the fermentation, the lower the

11 possibility of yielding alcohol because acetic acid becomes more potent when time passes by.

12 The optimum fermentation time is one week. Ethanol content is also affected by temperature—

13 the higher the temperature of the yield, the higher the concentration. The highest alcohol

14 concentration obtained from the experiments conducted was 70% with a density of

15 approximately 0.86 g/cm3.

16
17
18
19 Recommendations

20 More experimental studies focused on fermentation length should be done to determine

21 the exact length of time needed to produce the highest amount of alcohol. Future researchers

22 should also consider studying more banana varieties to know which variety is best to use for

20
1 producing alcohol. The alcohol yield should also be handled more carefully, and it is advisable

2 not to transfer it from one container to another to avoid contamination. Moreover, further

3 research needs to be done for alcohol purification.

21
 1 BIBLIOGRAPHY

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31 bacteria
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38

23
1 APPENDIX A

2 Concentration (In Weight %) of Ethanol-Water Mixture in Relation to Specific Gravity at

3 Various Temperature

24
1 APPENDIX B

25