USING SULFURIC ACID TO PRODUCE BIOETHANOL FROM FRUIT WASTE:

BANANA PEELS

ABSTRACT
Agricultural waste made of lignocellulosic materials, such as banana peels, has the potential to yield bioethanol, a
sustainable energy source. Our investigation verified the effectiveness of the acidic pretreatment method as a pre-
treatment for bioethanol production. We used the same H2SO4 hydrolysis method for the pretreatment method. Our
findings indicated that pretreating banana peels with acid reduced sugar content the most. The reducing sugars
were fermented for three days using an active strain of S. cerevisiae to produce bioethanol. The maximum absorp-
tion spectrum for the bioethanol was found at 280 nm using the potassium dichromate technique. The band in FT-
IR spectra confirmed the υ(OH) stretching vibration of ethanol.
Keywords - Bioethanol, Waste Banana peels, UV and IR.

1. GENERAL INFORMATION
The primary energy source in the world is fossil fuels, which are linked to of energy-related issues, in-
cluding climate change, global warming, uneven distribution within countries, and non-renewability. Bi-
oethanol, a clean fuel for combustion engines that is derived from plant materials, is a readily available
substitute; in 2008, its total consumption was over 65,000 million liters, and its use is growing rapidly, as
it replaced 5.4% of gasoline consumption in 2013 [1, 2]. However, deriving bioethanol from food sources
is not a viable alternative, because we have to choose between food and ethanol [3]. The chemical formula
of ethanol, generally known as alcohol, is C2H6O. It is a colorless, flammable, volatile liquid. Its density
is 0.789 g/cm3, its melting point is −114°C, its boiling point is 78.37°C, and its molar mass is 46.07
g/mole. It finds extensive usage in the industry as a fuel, solvent, and raw material for the synthesis of
other beneficial compounds. In addition, it is used as an antiseptic, heated in homes, and drunk as an
alcoholic beverage [4, 5]. It is an effective, economical, and secure food substitute. Dedicated energy
crops, industrial and municipal wastes, and agricultural or forestry leftovers are examples of this type of
biomass. The items that have been utilized earlier to create bioethanol include sugarcane bagasse, corn
curb, newspapers, sawdust, rice straw, wood, wheat, etc. One of the key characteristics of lignocellulosic
biomass is its carbohydrate content, which is necessary for the manufacture of ethanol. Cellulose (40–
50%), hemicellulose (25–35%), and lignin (15–20%) make up the biomass [6]. Out of the three compo-
nents, cellulose is the predominant polymer. Effective lignocellulosic biomass conversion to fermentable
sugars is the rate-limiting stage in the generation of ethanol [6, 7]. The kind of plant substantially deter-
mines the quantity of hemicellulose to be detected. Lignin, a complex polymer of aromatic alcohols known
as monolignols that sustain the plants, is the final part of the lignocellulosic biomass. The ultimate goal of
all pretreatments and hydrolysis procedures is to overcome the strength and compactness of the lignocel-
lulosic biomass in the ethanol production process [8]. There are two distinct steps involved in turning
cellulose polymer into ethanol. 1. Hydrolyzing cellulose to create glucose units; 2. Fermenting the sugars
obtained from the hydrolysis process with Saccharomyces cerevisiae to make ethanol. To produce the
sugars required for the hydrolysis and fermentation processes, pre-treatments are often used to separate
the mixed polymers of cellulose, hemicellulose, and lignin [9]. 15 billion metric tons of fossil fuels are
used annually. The three nations that utilize the most fossil fuels are China, the United States, and India.
According to the Global Material Flow Database (developed by UN Environment Program) By weight,
these nations use 54% of the fossil fuel produced worldwide. Bioethanol may be made inexpensively
available to consumers, boosting rural jobs and farmer income while lowering carbon emissions [10]. We
attempted to produce bioethanol from banana peels by pulping them using diluted sodium hydroxide,
hydrolysis, and acidic pretreatment with sulfuric acid. We chose banana waste because it was easily ac-
cessible and readily available, but we only used acid hydrolysis for the hydrolysis, which was then fol-
lowed by fermentation with S. cerevisiae. Banana peels are a great source of bioethanol, although the yield
depends on pre-treatment. This was determined by estimating the sugar content of the peels using Bene-
dict's solution [11].

2. MATERIALS AND METHODS

Banana peel waste is gathered from the neighbourhood fruit juice facility. The fruit skins were chopped
into little pieces to make it simpler to handle. They were exposed to both solar and overnight 60ºC oven
drying. For further examination, the dry substrate was pulverized, placed in a polyethylene bag, and kept
at 0°C.

2.1 PRETREATMENT OF WASTE BANANA PEEL

Pretreatment aims to lessen the fiber's crystal nature, compactness, and strength (cellulose). 200 ml solu-
tion of banana peel powder was steeped for 30 to 180 minutes, and the mixture was then autoclaved for
15 minutes at 120°C. The pretreated waste was filtered and filtrate was obtained.

2.2 HYDROLYSIS
Hydrolysis aims to further decompose polysaccharides into constituents of monosaccharides. A solution
that had been pretreated was combined with 10% sulfuric acid. The filtrate was mixed with sulfuric acid
in a 6:1 ratio, and the mixture was heated to 120°C for approximately 6 hours before being allowed to
cool. Before the fermentation process begins, the Benedict test is used to check the presence of reducing
and no reducing sugars.

2.3 BENEDICT TEST

It saves a great deal of time and work to confirm whether sugar is present before fermentation. The Ben-
edict test was used to figure out how much sugar was produced. 1 ml of the sample solution and 2 ml of
Benedict reagents were completely mixed in the test tube, and it was left in a boiling water bath for about
15 minutes. The presence of decreasing sugar is shown by the color changing from blue to green. The last
step in the synthesis of bioethanol is fermentation. With the aid of the enzymes zymase and invertase,
saccharomyces cerevisiae was utilized to hydrolyze the mono- and disaccharides generated into bioetha-
nol. It has been discovered that Saccharomyces cerevisiae has these enzymes. The hydrolyzed solution is
supplemented with yeast cells containing glucose. After the sample was centrifuged and the fermentation
process was allowed to continue for eight or nine days, the fermented bioethanol was distilled.

2.4 SAMPLE CHARACTERIZATION

To determine its unique qualities, the produced bioethanol was subjected to characterization procedures.
To determine the surface plasmon resonance (SPR) band for optical qualities, the absorption spectra was
recorded using a UV-Vis spectrometer. The method of FTIR Spectroscopy involves determining how an
infrared radiation interacts with a sample, which may be solid, liquid, or gaseous. It detects the strengths
of these absorptions as well as the frequencies at which the sample absorbs. Because various frequencies
of radiation are absorbed by chemical functional groups, the frequencies may be used to determine the
chemical composition of the sample.
 3. RESULTS AND DISCUSSION
The investigation's findings demonstrated that a sizable amount of ethanol was created by the fermented
banana peels. To producing ethanol, the solution was stored for three to four days. Regularly record any
changes. As the quantity of yeast increases, the amount needed for the fermentation process drastically
lowers. The Bioethanol estimation done by potassium dichromate method (Five different test tubes were
taken and to each test tube ethanol is added in the increasing concentration and then 2ml of potassium
dichromate solution is added to each test tube and shaked well. UV-Vis Spectroscopy. the absorption peak
or biodiesel is around 280 nm this is the value of λ max of bioethanol as shown in Figure 1.

Figure 1 – The absorption peaks for bioethanol

3.1 INFRARED SPECTRAL ANALYSIS

The infrared absorption spectra of the investigated bioethanol are shown in figure 2. Inspection of the IR
spectra and the bands frequencies data shows the presence of a broad band at 3498-3496 cm-1 which
corresponds to the stretching vibration of the OH group. The broad shape is usually attributed to the fact
that the molecules containing the OH group. The bands in the 2821–2819 cm-1 region are caused by the
stretching vibration of aliphatic C–H bonds (alkyl group) [1212].

Figure 2. The Infrared of bioethanol

4. CONCLUSION
To determine whether it would be feasible to produce bioethanol from banana peels as a source of ligno-
cellulosic biomass, this study used three different pretreatment techniques as the first step in the experi-
mental design: acidic, alkaline, and water pretreatments. These techniques produced varying concentra-
tions of reducing sugar after hydrolysis with sulfuric acid; Benedict's test confirmed that pretreatment with
acid produced the highest reducing sugar concentration. However, alkaline pretreatment achieved the best
conversion of reducing sugars to ethanol. The outcome unequivocally shows that every pretreatment
method has benefits of its own. Pretreatment with water alone, without any chemical augmentation, often
produced poor results in the glucose and ethanol studies for Bioethanol Production from Banana Peels.
This study has established that the most effective way to make ethanol from banana waste is to use an
alkaline pretreatment, followed by acid hydrolysis, and then S. cerevisiae for the fermentation process. In
general, additional enzymatic approaches should be applied for the hydrolysis because acid hydrolysis is
quite extremely costly. More particular ways to increase the production of bioethanol are therefore neces-
sary.

REFERENCES
[1] T. H. Kim, F. Taylor, and K. B. Hicks, "Bioethanol production from barley hull using SAA (soaking
in aqueous ammonia) pretreatment," Bioresource technology, vol. 99, pp. 5694-5702, 2008.
[2] W. K. El-Zawawy, M. M. Ibrahim, Y. R. Abdel-Fattah, N. A. Soliman, and M. M. Mahmoud, "Acid
and enzyme hydrolysis to convert pretreated lignocellulosic materials into glucose for ethanol produc-
tion," Carbohydrate polymers, vol. 84, pp. 865-871, 2011.
[3] S. Sakai, Y. Tsuchida, S. Okino, O. Ichihashi, H. Kawaguchi, T. Watanabe, et al., "Effect of ligno-
cellulose-derived inhibitors on growth of and ethanol production by growth-arrested Corynebacterium
glutamicum R," Applied and environmental microbiology, vol. 73, pp. 2349-2353, 2007.
[4] N. Curreli, M. B. Fadda, A. Rescigno, A. C. Rinaldi, G. Soddu, F. Sollai, et al., "Mild alkaline/oxi-
dative pretreatment of wheat straw," Process biochemistry, vol. 32, pp. 665-670, 1997.
[5] B. Yang and C. E. Wyman, "Effect of xylan and lignin removal by batch and flowthrough pretreat-
ment on the enzymatic digestibility of corn stover cellulose," Biotechnology and bioengineering, vol. 86,
pp. 88-98, 2004.
[6] K. A. Gray, L. Zhao, and M. Emptage, "Bioethanol," Current opinion in chemical biology, vol. 10,
pp. 141-146, 2006.
[7] A. E. Farrell, R. J. Plevin, B. T. Turner, A. D. Jones, M. O'hare, and D. M. Kammen, "Ethanol can
contribute to energy and environmental goals," Science, vol. 311, pp. 506-508, 2006.
[8] C. E. Wyman, "Potential synergies and challenges in refining cellulosic biomass to fuels, chemicals,
and power," Biotechnology progress, vol. 19, pp. 254-262, 2003.
[9] A. Endo, T. Nakamura, A. Ando, K. Tokuyasu, and J. Shima, "Genome-wide screening of the genes
required for tolerance to vanillin, which is a potential inhibitor of bioethanol fermentation, in Saccharo-
myces cerevisiae," Biotechnology for Biofuels, vol. 1, pp. 1-6, 2008.
[10] A. Hendriks and G. Zeeman, "Pretreatments to enhance the digestibility of lignocellulosic biomass,"
Bioresource technology, vol. 100, pp. 10-18, 2009.
[11] C. Cardona, J. Quintero, and I. Paz, "Production of bioethanol from sugarcane bagasse: status and
perspectives," Bioresource technology, vol. 101, pp. 4754-4766, 2010.
[12] C. K. Phwan, K. W. Chew, A. H. Sebayang, H. C. Ong, T. C. Ling, M. A. Malek, et al., "Effects of
acids pre-treatment on the microbial fermentation process for bioethanol production from microalgae,"
Biotechnology for biofuels, vol. 12, pp. 1-8, 2019.