IOSR Journal of Applied Chemistry (IOSR-JAC)

e-ISSN: 2278-5736.Volume 17, Issue 5 Ser. 1 (May 2024), PP 07-13

www.iosrjournals.org

Production And Characterization Of Bioethanol From

Cassava Peels
Irene Akinyi Odongo1, Elly Tetty Osewe1, Sylvia Opiyo1.
1
(Department Of Physical And Biological Sciences, Murang’a University Of Technology, Kenya)

Abstract
Majority of rural Africans use inefficient energy sources like wood and fossil fuels, which are not environmentally
friendly and hazardous to human health. Bioethanol is an alternative clean source of energy that can be obtained
from biomass and other agricultural wastes. Producing bioethanol from agricultural wastes cleans up the
environment as it prevents the cutting of trees for wood fuel, thus solve the problem of de- forestation. It also
gives farmers an additional source of income from otherwise wasted agricultural materials, reduces indoor
pollution, and is more cost-effective due to its higher calorific value. This research focused on characterization
of cassava (Manihot esculenta Crantz) peel bioethanol . Bulk fermentation of bioethanol was conducted under
optimum fermentation conditions, which were determined through Taguchi optimization. The experimentation
involved the utilization of cassava (Manihot esculenta Crantz) peel pretreated with 0.5 M sulphuric acid.
Saccharomyces cerevisiae were employed as the yeast source to ferment the acid-pretreated cassava peels. Bulk
production was conducted to ensure a sufficient quantity of bioethanol was generated for characterization . The
volume of the substrate was raised to 500 ml, and 45 g of yeast was added to the substrate media to start the
scaled-up production. In addition, the fermentation period was 18 hours, precise temperature of 40 oC and a pH
of 4.5. After the production the produced bioethanol was characterized to establish the following parameters i.e.
the density, specific gravity, ethanol concentration, ash content sulphur content, calorific value, pH and the
electrical conductivity.The bioethanol produced was characterized and the following parameters registered; The
bioethanol yield was 200 L/ton of cassava peels bioethanol, density of 0.8763 g/cm3, specific gravity of 0.8779
g/cm3, ethanol concentration of 73.63 %v/v, ash content of 0.005, sulphur content of 0.0788, flash point of 17 oC,
kinematic viscosity of 3.677 centistrokes (cst), the calorific value of 21.89 MJ/kg, pH of 4.286, and conductivity
of 8.81 𝜇s/m.
Keywords: Cassava peel; Bioethanol; Taguchi; Fermentation; Characterization; Saccharomyces cerevisiae
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Date of Submission: 24-04-2024 Date of Acceptance: 04-05-2024
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I. Introduction
The reliance on fossil fuels as the dominant source of energy has resulted in many detrimental
environmental consequences1. Emissions of hazardous particles and gases play a significant role in climate
change. Pollutant gases cause a variety of health and developmental defects in children 2. Nowadays, bioethanol
is the most utilized liquid biofuel on the market. 3. Therefore, biofuels are alternative sources of clean and
affordable energy4. The manufacture of bioethanol uses many biomass sources 5. Many countries have set lofty
goals for replacing petroleum-based fuels with them 6. Before fermentation, lignocellulose must undergo
pretreatment. Various methods are available such as using physical, chemical, and physicochemical methods and
biological agents like Aspergillus niger, A. oryzae, Humicola insolens and Trichoderma reesei, which deconstruct
the lignocellulose structure 7,8. Although fermentation is a natural process, microorganisms are still needed for
the conversion of C6H12O6 into C2H5OH or lactic acid, or many other products, where numerous industrial yeasts,
including, Saccharomyces cerevisiae are employed for these purposes 9. It is the primary fermentative strain
employed in industries producing sugar-based biofuels. Pretreatment transforms the cellulosic material or slurry
into fermentable sugars, making it suitable for either enzymatic or acidic hydrolysis. Saccharomyces cerevisiae
is used in fermentation to convert C6H12O6 into C2H5OH under anaerobic conditions and at a specific temperature
9
. The yeast cells directly metabolize a sugar like glucose through the glycolysis pathway in order to produce
energy for biosynthesis. For every mole of glucose consumed, the fermentation process' total reactions provide
two moles of ethanol and carbon dioxide 10,11

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 Production And Characterization Of Bioethanol From Cassava Peels

12 11
Scheme 1: Yeast fermentation of glucose into ethanol ,

The summary of the fermentation process is shown in equation (1) below;

(1)
C6 H12 O6 (l) → 2C2 H5 OH(l) + 2CO2 ( 𝑔)
Factors that affect bioethanol yield include; pHs, fermentation time, yeast: substrate ratio, and
temperature. Based on the results, the greatest bio-ethanol yield was 45.50 % at the same time the fermentation
temperature was 35 oC, the yeast concentration was 10 %, the C6H12O6 concentration was 100 g/L, and the
fermentation time was 6 days 13. Eighty three percent of C2H5OH was obtained from cassava peel using Zymonas
mobilis14. Egbosiuba 13 characterized the obtained bioethanol and obtained that the water (H 2O) content of the
bioethanol produced was 0.0021 ppm, sulphur content was 0.0003 %, which was lower than the ASTM maximum
standard fuel sulphur value of 0.05 %.The flash point of the bioethanol produced was 14.2 oC. Bioethanol
generated had a specific gravity of 0.785 kg/L at room and at standard temperature was 0.750 kg/L.

II. Material And Methods

Yeast (Saccharomyces cerevisiae) was purchased from a local supermarket. Pearl millet was obtained
from a local market, Buffer tablets pH 4 and 7, NaOH 99.9%, and H2SO4 98% of analytical grades were obtained
from Sigma Aldrich Chemical distributors, Nairobi Kenya.

Sample collection and preparation

The cassava used in this study was obtained from a farmer in Kodiaga and its geographical coordinates
is 0°05'55.7" North and 34°26'14.8" East situated in North Gem, Siaya County, Kenya. The cyanic cassava was
washed with water to remove soil pollutants, fertilizers and other foreign substances associated with biomass
waste and then peeled off. The peels were sun-dried for one week to remove the moisture and finally milled to
obtain cassava peel powder.

Sulphuric acid pretreatment

Using Yussouff 15 method with some modifications, acid hydrolysis was carried out by adding 200 g of
cassava peels powder to 1000 ml of 0.5 M sulphuric acid in a 1000 ml beaker. The samples were heated for 2
hours in a water bath at 98 °C, sterilized in an autoclave for 15 minutes at 121°C, allowed to cool, and then filtered

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 Production And Characterization Of Bioethanol From Cassava Peels

through No. 1 whatman filter paper using vacuum filtration method. The sample was stored in the refrigerator at
4 oC and later used for bioethanol production.

Determination of the percentage sugar content

After cassava was hydrolyzed, the quantity of total sugar in the hydrolysate was established using
refractive index. To do this, a drop of cassava hydrolysate was applied to the glass slide a graduated hand
refractometer model REF107 and the brix reading was expressed as a percentage. Following AOAC (2000), a
hand refractometer model REF107 was used to calculate the brix (%) 16.

Production of bioethanol
For the scaled production, 1200 ml of sulphuric pretreated cassava filtrate was poured into a 2000 ml
beaker. The sample was then placed on a magnetic stirrer and the filtrate pH was adjusted to 4.5 using 825 ml of
sodium hydroxide. Subsequently, 500 ml of the adjusted filtrate was poured into four 500 ml Erlenmeyer flasks
with 45 g of yeast added to each flask. These flasks were then placed in a water bath set at 40 oC for six hours.
Overnight, the samples were placed in an incubator set at 40 oC to continue fermenting for the next 12 hours,
totaling the fermentation time to 18 hours. Following this, the samples were immediately distilled off for further
processing. The fermented liquid was poured into a round-bottomed flask, placed on a heating mantle connected
to a distillation column, and immersed in running water. The distillate was collected in a vial placed to the opposite
end of the distillation column at 78 oC. This was done for each fermented broth using the method indicated by
Oyeleke 17.

Estimation of bioethanol yield

The yield of bioethanol can be stated in a variety of ways, such as mL/g of substrate, g/g of substrate,
mL/L of solution, g/L of solution, etc. In order to improve comparability, the bioethanol yield unit was changed
to the same unit. The unit was changed in this study into L bioethanol/ton for cassava peel substrate based on the
following assumptions in the equation(2) and (3) 18.

𝑔 𝑏𝑖𝑜𝑒𝑡ℎ𝑎𝑛𝑜𝑙 𝑔 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒
=
1 𝑙𝑖𝑡𝑟𝑒 𝑓𝑖𝑛𝑎𝑙 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 1 𝑙𝑖𝑡𝑟𝑒 𝑖𝑛𝑖𝑡𝑖𝑎𝑙𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 (2)

𝐿 𝑏𝑖𝑜𝑒𝑡ℎ𝑎𝑛𝑜𝑙
= (3)
𝑡𝑜𝑛 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒

Determination of density, specific gravity and ethanol concentration

The density meter model DMATM 4100 Instrument was calibrated using water as calibration standards.
Using a syringe the sample was slowly injected into the glass oscillator until it was completely filled and care
was taken to ensure that no air bubbles was present. The readings were taken once constant temperature of 20.0
±0.3oC was reached.

Determination of ash content

The ASTM D1998 method was used to establish the amount of ash in the biomass samples 1g of each
sample was placed in a pre-weighed crucible, which was then placed into a desiccator to dry after being burned
at 550 oC for 1 hour in a muffle furnace for ashing process.

Ash content was calculated using Equation (4):

𝑊2 − 𝑊0
𝐴𝑠ℎ 𝑐𝑜𝑛𝑡𝑒𝑛𝑡 (%) = × 100%
𝑊1 − 𝑊0 (4)

Where: W1 = weight of the container + biomass sample before burning

W0 = Weight of the container
W2 = weight of the container + biomass sample after burning

Determination of sulphur content

Using Ismail 19 methods with some modifications 20 ml of 1:1 hydrochloric acid was added to 20 ml of
bioethanol in a beaker, and the mixture was stirred. This step converted the sulfur in the bioethanol to sulphuric
acid. The beaker was then placed on a sand bath and heated for 30 minutes, evaporating the excess water and
ethanol and concentrating the sulphuric acid. The solution was filtered using a filter paper, and the filtrate was
collected in another beaker, while the residue on the filter paper was discarded. 20 ml of barium chloride solution
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 Production And Characterization Of Bioethanol From Cassava Peels

was added to the filtrate, and the mixture was stirred well, precipitating the sulphuric acid as barium sulfate. The
mixture was allowed to stand until the precipitate settled at the bottom of the beaker. If no precipitate formed, it
indicated that the sulfur content was very low or negligible. The mixture was filtered again using a filter paper,
and the precipitate was collected on the filter paper. The precipitate was washed with distilled water to remove
any impurities. The filter paper with the precipitate was transferred to a pre-weighed crucible or beaker.
Additionally, an empty filter paper was weighed to use as a blank. The filter paper with the precipitate and the
blank were ashed until they turned white, removing the organic matter and leaving only the barium sulfate. The
ashed samples and the blank were weighed, and the weight of the blank was subtracted from the weight of the
sample to determine the weight of the barium sulfate. The weight of sulfur in the sample was calculated using
equation (5):
𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑢𝑙𝑝ℎ𝑢𝑟 = 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑏𝑎𝑟𝑖𝑢𝑚 𝑠𝑢𝑙𝑝ℎ𝑎𝑡𝑒 × 233.3906 (5)
Where,
32.06 is the atomic weight of sulphur in g/mol
233.39 is the molecular weight of barium sulphate in g/mol
The percentage of sulfur in the sample was calculated using equation (6)
𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑢𝑙𝑝ℎ𝑢𝑟
𝑃𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑜𝑓 𝑠𝑢𝑙𝑝ℎ𝑢𝑟 = × 100%
𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑏𝑖𝑜𝑒𝑡ℎ𝑎𝑛𝑜𝑙 𝑠𝑎𝑚𝑝𝑙𝑒 (6)

Determination of flash point

The flash point of bioethanol was determined using the Pensky Martens (ASTMD93) method. The test
sample was placed in a metal cup in such a way that it just touched the prescribed mark on the inside of the cup.
The cover was then be positioned on the cup. The Bunsen burner was used to deliver heat to the apparatus's lower
side. The heating was set to provide a 7 0F per minute temperature rise, and the sample was constantly agitated.
As the temperature of the flash approaches, the injector burner lights up and then injected in the sample at
approximately 12-second intervals until a distinct flash is visible within the container and the injector burner is
turned off. A thermometer was used to determine the close flash point at this moment 20.

Determination of viscosity
The Ostwald viscometer was first taken and thoroughly rinsed with water. Subsequently, the apparatus
was dried in the oven. It was filled with the test then partially sample (bioethanol) from the lower end of the tube.
The liquid was introduced into the viscometer and drawn up to the upper bulb of the tube. The level was
maintained stationary by holding it with a finger. The time taken by the sample to descend from the upper marked
level to the lower marked level was carefully observed. This process was repeated twice more, and readings were
recorded each time 21.

Determination of calorific value

Calorimeter model BCY-1A was used to measure the calorific value of the fuel. A crucible was inserted
into the bomb after adding of 1g of the fuel. The bomb was filled with oxygen and placed inside the jacket before
the fuse was connected with a nichrome wire. The jacket was filled with 2000 ml of water and the calorimeter
was put together. A thermometer was used to check the temperature once every minute until it reached a steady
value 22.

Determination of pH
The bioethanol was tested using a pH meter biobase model 930 by dipping the pH meter electrode into
the sample until it showed a stable scale reading on the pH meter 23.

Determination of electrical conductivity

The instrument utilized was a digital electrical conductivity meter model 950. 30 ml of bioethanol was
transferred into a 100 ml beaker. The electrode was submerged up to the point of censorship. After a stable reading
was acquired, the meter was turned on to the necessary calibrated units, and readings was done in triplicate 24.

III. Results And Discussion

Characterization of cassava peel bioethanol
In this study, instead of the typical expression of bioethanol yield as grams of bioethanol per gram of
substrate, the yield was measured as liters of bioethanol per fresh cassava, to facilitate comparison with previous
research findings. In this study, the yield of bioethanol was established using equation 2 and 3 above. The yield
of bioethanol was 200L/ton of cassava peels bioethanol was close to 160 L/ton reported by Dinata and Kartawiria
18
. Density is a crucial characteristic of fuel that plays a significant role in determining its energy content and

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 Production And Characterization Of Bioethanol From Cassava Peels

practical utility. Ethanol combustion efficiency is crucial for optimizing the quantity of energy converted from
the fuel to heat. Incomplete combustion reduces cooking efficiency and results to energy loss. The key to burning
ethanol efficiently is to achieve full combustion, which reduces the amount of unwanted byproducts like carbon
monoxide produced and instead converts the fuel to CO 2 and H2O. The density of the produced bioethanol was
0.8763g/cm3 with a specific gravity of was found to be 0.8779g/cm3 (Table 1), which was lower than 0.9970
g/cm3 and 0.955 g/cm3 specific gravities reported by Omoruyi 25 and Ogunsuyi 26 respectively.

Table 1: Properties of bioethanol produced from sulphuric acid pretreated cassava peels
ASTM Standards for
S/N parameter Units Experimental results
Bioethanol
1 Density @ 20 OC g/cm3 0.8763 0.750-850
2 Specific gravity g/cm3 0.8779 0.87
3 Concentration of EtOH % v/v 73.63 -
4 Ash 0.005 -
Kinematic Viscosity @ 20
5 O Cts/sec 3.6765 5.0 max
C
7 Sulphur content % wt 0.0788 0.05
O
8 Flash point (closed cup) C 17 18.60
9 pH a.u 4.286 -
10 Calorific value MJ/kg 26.89-29.70 21.89 MJ/kg
11 Electrical conductivity µS/m 8.81 500max

The concentration of ethanol refers to the amount or proportion of ethanol present in a solution. The
concentration of ethanol in the produced bioethanol was 73.63 %v/v, which was higher than 45.50 % reported by
Egbosiuba 13.The ash content of the produced bioethanol was 0.005, which can be considered very negligible.
The ash content value was comparable to that previously reported 27. Ash buildup can decrease airflow and impair
efficient combustion. Ash and other byproducts may be produced when ethanol is not completely burned. A
portion of these byproducts might contain contaminants that worsen indoor air quality, which has an impact on
the environment and public health
Viscosity is a measure of a fluid's resistance to progressive deformation brought on by shear or tensile
stress. When adjacent particles in a fluid collide while moving at different speeds, viscosity is produce 21. The
atomization of the fuel during combustion is impacted by the fuel's viscosity. Increased viscosity can lead to
inefficient combustion and poor atomization, which would reduce the burning efficiency and increase emissions.
Low kinematic viscosity of ethanol produces less emission during combustion, especially when it comes to
particle matter. Reduced production of smoke and other pollutants as well as more thorough combustion can result
from the fuel's fine atomization. The produced bioethanol had a kinematic viscosity of 3.6765 cst (Table 1) which
was lower but within the ASTM standards than the kinematic viscosity of 1.21×10 3 cst previously reported 13, but
was within the ASTM standards.
The amount of sulphur in the produced bioethanol was 0.0788012 % which was close to the one reported
by Umeuzuegbu 28 of 0.06 % but lower than the sulphur content of 1.58 reported by Suleiman 20. The amount of
sulphur in bioethanol can affect how well it burns. Incomplete combustion of sulphur can produce sulphur dioxide
(SO2). The sulphur level of bioethanol is typically a cause for worry. A high sulphur level may cause partial
combustion by interfering with the burning process. In addition to producing less energy, incomplete combustion
releases hazardous byproducts such carbon monoxide (CO) and particulate particles.
The flash point of the produced bioethanol was 17 oC which falls closely within the range of ASTM
standards for combustible and flammable liquids but lower than the flash point of 46 oC reported by Ogunsuyi 26.
Flame and combustible-grade liquids are fuels with a flash point below 37.8 OC (ASTM, 2011). Due to its volatile
nature, bioethanol vapor and oxygen may combine to create a combustible mixture. Because of this, bioethanol
evaporation into the air occurs more quickly at higher concentrations, leading to increased combustibility. Higher
burning efficiency is preferred to minimize fuel use, guarantee lower emissions, and enhance fuel’s efficiency.
The low flash point of 17 oC of ethanol implies that ethanol can quickly combine to create a combustible mixture,
which can ignite spontaneously.
The pH of the produced bioethanol was 4.286, which was higher and thus less acidic than a pH of 2.85
reported previously 29. In terms of acidity, ethanol is rather a weak acid. When contaminants are present in the
ethanol, they may behave as bases or acids, changing the solution's overall acidity. Pollutants or impurities can
significantly impact the suitability of fuel for various applications. Extremely low pH levels can corrode fuel
storage tanks and transportation infrastructure, potentially rendering them unfit for use in certain environment.
The calorific value of fuel bioethanol was 21.89 MJ/kg, which was a bit lower than the 29.78 MJ/kg
calorific value of ethanol reported Nwufo 30. However, the calorific value can be increase by adding the number
of Carboxy Methyl Cellulose (CMC) 31. The presence of carbon and hydrogen content elements in CMC has the
potential to enhance the fuel's calorific value. This is due to the combustion process, where carbon interacts with

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 Production And Characterization Of Bioethanol From Cassava Peels

oxygen, resulting in the generation of heat 31. The calorific value of fuel bioethanol reveals the amount of energy
one can obtain by burning a specific mass of bioethanol. A higher calorific value implies that the fuel can release
more energy when combusted, making it more efficient for use as a source of heat.
The produced bioethanol had an electrical conductivity of 8.81𝜇s/m which was within the ASTM
standards (Table 1). The electrical conductivity was lower than the electrical conductivity of 329 𝜇s/m, 330
𝜇s/m, 230 𝜇s/m,300 𝜇s/m ,150 𝜇s/m obtained from the following varieties of cassava TMS 92B/00068 ,TMS
91/02324 ,TMS 92B/0061 ,TMS 98/0505 ,TMS 98/058 respectively reported by Ademiluyi 24. Fuel ethanol's
electrical conductivity is a crucial factor to consider when assessing the quality of the fuel. Electrochemical or
galvanic corrosion is more likely to occur in high-conductive fluids due to the correlation between conductivity
and the presence of corrosive ions like chlorine 32. Higher purity bioethanol typically has lower electrical
conductivity. Bioethanol boasts a commendably low electrical conductivity, attributed to its pure composition
resulting from fermentation, where the absence of ionic components prevents an increase in its conductivity.

IV. Conclusion
The cassava peels not only satisfies the ASTM legal requirements but also provide advantages for the
environment. Cassava peel bioethanol can therefore be suggested as a practical alternative fuel source for a range
of applications, supporting sustainable energy practices and lowering reliance on fossil fuels, based on the
empirical evidence provided in this work. The 200litres per tonne of bioethanol was obtained at the optimum pH
of 4.5, temperature of 40 oC, 45 g of yeast, glucose concentration of 14.5 %, fermentation period of 18 hours. The
properties of the cassava peels bioethanol had the following characteristics density of 0.8763 g/cm3, specific
gravity of 0.8779 g/cm3, ethanol concentration of 73.63 %v/v, ash content of 0.005, sulphur content of 0.0788,
flash point of 17 oC, kinematic viscosity of 3.677 centistrokes (cst), calorific value of 21.89 MJ/kg, pH of 4.286,
and conductivity of 8.81 𝜇s/m.

V. Recommendation
In order to enhance the production of bioethanol under the specified conditions of pH 4.5, yeast quantity
of 45 g, temperature of 40 °C, and fermentation duration of 18 hours, it is very important to maintain pH of the
fermentation media throughout the process this will ensure optimal activity of the yeast. Quality yeast strains
should be utilized, with careful attention to the viability of the yeast strain that used during fermentation.
Fermentation temperature to be strictly maintained at 40 °C for optimal activity of the yeast, and process of
fermentation to be monitored closely to adjust parameters as needed.

Future works
Bioethanol, while meeting biofuel requirements, there is needs to improve its calorific value.
Carboxymethyl Cellulose (CMC) can increase ethanol's calorific value by adding more flammable material and
increasing the carbon-to-hydrogen ratio. CMC also improves fuel atomization and combustion efficiency,
allowing for more thorough mixing of fuel with air .

Declaration of conflict of interest

The authors hereby declare no conflict of interest in publishing this paper.

Acknowledgement
I would like to extend my appreciation to CIST Africa Limited for providing financial support, which
significantly contributed to the successful completion of this work. I extend my heartfelt appreciation to Prof.
Crispin Kowenje of Maseno University Kenya, for his invaluable contributions to this work. My appreciation also
goes to Mr. Langat Johnston Kibet, a research scientist at Kenya Industrial Research and Development Nairobi
Kenya for offering me a laboratory space and guiding me during characterization.

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