Research Methodology Project 2019

MEKELLE UNIVERSITY
EIT-M
DEPARTMENT OF CHEMICAL ENGINEERING
STREAM: BIOCHEMICAL ENGINEERING
COURSE TITLE: RESEARCH METHODS AND PROPOSAL WRITING
COURSE CODE: CHEG251
PROJECT TITLE: PRODUCTION OF BIOETHANOL FROM CORN COB
PREPARED BY:
NAME ID NUMBER
1) Teklemariam Negash Eitm/ur83233/07
2) Marta Tesfaye Eitm/ur82728/07
3) Abraham Legesse EIT-M/UR/161456/2006
4) Haftom Mezgobo Eitm/ur82426/07
5) Firaol Kasaye EIT/UR1511/05
6) Yordanos Alem EIT-M/UR/263248/2006
7) Shewit Awetehey EIT-M/UR/262941/2006
8) Ytbarek Shshaye Eitm/ur83450/07
9) Asefa Abraha EIT-M/UR/161603/06
10) Wolega Wodamo Eitm/ur83356/07
11) Brhanu Belay Eitm/ur81905/07
12) Fana Teka EIT-M/UR/261966/2006
13) Meseret Eshetu Eitm/ur82816/07
Submission Date: 12/06/2011E.C

Submitted to: Inst.YALEMBRHAN D. (MSc) MEKELLE, ETHIOPIA
PRODUCTION OF BIOETHANOL FROM CORN COB
ACKNOWLEDGMENT
First of all we would like to thank the Almighty God for giving us the strength and patience for
the successful accomplishment of this project.
Next we would like to take this opportunity for expressing our deep and sincere gratitude and
thankfulness to our dearest inst. Yalembrhan D. (MSc), for his advices, suggestions, guidance
and giving enough time to complete our project.
In closing, we are deeply grateful to our class mate for their willingness and kindness to share
ideas with us about their and ours projects.
Department of Chemical Engineering Page i

TABLE OF CONTENTS
ACKNOWLEDGMENT ............................................................................................................. i
LIST OF FIGURES .................................................................................................................. iv
LIST OF TABLES ......................................................................................................................v
LIST OF ABRREVATIONS AND ACRONYEMS .................................................................. vi
ABSTRACT ............................................................................................................................ vii
1 INTRODUCTION....................................................................................................................1
1.2 Background .......................................................................................................................1
1.2 Statement of the Problem ...................................................................................................2
1.3 Objective ...........................................................................................................................3
1.3.1 General objective.........................................................................................................3
1.3.2 Specific objective ........................................................................................................3
1.4 Significance of the project .................................................................................................3
2 LITERATURE REVIEW .........................................................................................................4
2.1 Background .......................................................................................................................4
2.1.1 Overview of maize production in Ethiopia ...................................................................4
2.2 Bioethanol and its characteristics .......................................................................................6
2.3 Bioethanol feedstock’s .......................................................................................................7
2.3.1 First generation feedstocks ..........................................................................................7
2.3.2 Second generation feedstocks ......................................................................................8
2.3.3 Third generation feedstocks ....................................................................................... 11
2.4 Corncobs as a Feedstock of Bioethanol ............................................................................ 11
2.5 Bioethanol Production Process ......................................................................................... 12
2.5.1 Biochemical Conversion ............................................................................................ 12
2.5.2 Thermochemical Process ........................................................................................... 18
2.6 Factors Affecting Fermentation ........................................................................................ 19
3 METHODOLOGY ................................................................................................................. 21
3.1 Materials .......................................................................................................................... 21
3.1.1 Characterization of corncob ....................................................................................... 21
3.2 Corn cob Preparation ....................................................................................................... 22
3.3 Acid pretreatment ............................................................................................................ 23
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3.3 Dilute Acid Hydrolysis .................................................................................................... 23

3.4 Fermentation .................................................................................................................... 24
3.5 Bioethanol Separation ...................................................................................................... 24
4 RESULTS AND DISCUSSION ............................................................................................. 26
4.1 Characterization of corncob ............................................................................................. 26
4.2 Effect of Acid hydrolysis on bioethanol yield................................................................... 27
5 MATERIAL AND ENERGY BALANCE .............................................................................. 29
5.1 Material and energy balance overview ............................................................................. 29
5.2 Material Balance .............................................................................................................. 29
5.3 Energy balance ................................................................................................................ 36
6 SIZING OF MAJOR EQUIPMENTS ..................................................................................... 42
7 PRELIMINARY ENGINEERING ECONOMIC ANALYSIS ................................................ 50
7.1 Total Capital Investment .................................................................................................. 50
7.2 Total production cost ....................................................................................................... 51
7.3 Economic evaluation ........................................................................................................ 53
8 CONCLUSION AND RECOMMENDATIONS .................................................................... 55
8.1Conclusion........................................................................................................................ 55
8.2 Recommendation ............................................................................................................. 56
REFERENCE ........................................................................................................................... 57
APPENDIX .............................................................................................................................. 59
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LIST OF FIGURES
Figure 2.1 Lignocellulose structure, showing cellulose, hemicellulose and lignin fractions ..........9
Figure 3.1 corn cob and grounded corn cob ............................................................................... 23
Figure 3.2 Production process of bioethanol from corn cob ....................................................... 25
Figure 4.1 Concentration of standard glucose and its absorbance ............................................... 27
Figure 4.2 Total reducing sugar and the bioethanol yield from corn cob ................................... 28
Figure 5.1 Production flow diagram of bioethanol from corn cob .............................................. 30
Figure 5.2 Material balance on molecular sieve ........................................................................ 31
Figure 5.3 Material balance on distillation ................................................................................ 32
Figure 5.4 Material balance on fermenter .................................................................................. 33
Figure 5.5 Material balance on filter ......................................................................................... 34
Figure 5.6 Material balance on hydrolysis reactor...................................................................... 35
Figure 5.7 Material balance on dryer ........................................................................................ 36
Figure 5.8 Energy balance on dryer ........................................................................................... 37
Figure 5.9 Energy balance on hydrolysis reactor ....................................................................... 37
Figure 5.10 Energy balance on fermenter .................................................................................. 38
Figure 5.11 Energy balance on distillation column .................................................................... 39
Figure 5.12 Energy balance on molecular sieve ......................................................................... 40
Figure 6.1 McCabe-Thiele diagrams to determine number of stage ........................................... 46
Figure 6.2 Flow sheet of bioethanol production from corn cob .................................................. 49
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LIST OF TABLES
Table 2.1 Major Maize Producing Administrative Zones of Ethiopia ...........................................6

Table 2.2 Physico-chemical characteristics of bioethanol ............................................................6
Table 4.1 Proximate analyses of the corncob sample ................................................................. 26
Table 4.2 chemical composition of corncob sample ................................................................... 26
Table 6.1 Vapor liquid equilibrium data of bioethanol water solution ........................................ 45
Table 7.1 purchased cost of equipment ...................................................................................... 50
Table 7.2 estimation of fixed capital investment ........................................................................ 51
Table 7.3 Total cost of raw materials ......................................................................................... 52
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LIST OF ABRREVATIONS AND ACRONYEMS
Cp Specific heat capacity

E Purchased Equipment cost
DC Direct cost
FCI Fixed capital investment
IC Indirect cost
MPa Mega Pascal
PH potential of hydrogen
S. cerevisiae Saccharomyces cerevisiae
TCI Total Capital Investment
TDC Total direct cost
TPC Total production cost
TRS Total Reducing Sugar
WC Working Cost
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ABSTRACT
The objective of the study was production of bioethanol from corncob which in effects to
minimize energy cost and substituting nonrenewable energy by using renewable resources. The
production process was carried in four main stage such as pretreatment, hydrolysis (first and
second stage), fermentation and distillations. The first stage diluted acid hydrolysis was used as
chemical pretreatment stage and the process variables were fixed at the best optimum condition.
Before the first stage hydrolysis process was carried out, the corn cob was dried by dryer at 70oC
temperature for 24 hr. After hydrolysis process sugar content of the hydrolyzate was quantified
using quantitative benedict reagent solution. Fermentation of the hydrolyzate was performed
using Saccharomyces cerevisiae at 30oC temperature, pH5.0 and 72hr fermentation time. In the
process, it was observed that bioethanol concentration decreased with increase in acid
concentration, hydrolysis time and fermentation time. The maximum result was obtained with
distilled water hydrolysis for 4 h and 24 h fermentation. Under this condition maximum
bioethanol concentration production was 7.398g/L, a satisfactory result as compared with
literature data.
Keywords: Bioethanol, corn cob, diluted-acid hydrolysis, and Saccharomyces cerevisiae
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1 INTRODUCTION
1.2 Background
Energy consumption has increased during the last century due to the world population
development and growth. Nowadays, the increasing problem of the CO2 emissions due to energy
consumption, besides to the future petroleum scarcity, has strengthened the interest in
alternative, nonpetroleum-based sources of energy (Montoya G, 2014).
One of the potential options to solve the environmental and energetic problems is the use of bio-
ethanol. This is a renewable fuel, which avoids the negative environmental impacts generated by
petroleum-based fuels. (Montoya G, 2014).
The importance of ethanol as a clean and safe transportation fuel has increased with the
anticipated shortage of fossil fuel reserves and increased air environmental sustainability.
However a dramatic increase in ethanol production using the current starch-based technology
may not be practical because it will compete for the limited agricultural land needed for
food and feed production, thus affecting food security.
Lignocellulosic biomass is a cheap, renewable, abundantly available resource (Hansen et

al., 2005), and its conversion to glucose and other fermentable sugars has been
considered to be an attractive route for ethanol production (Cao et al., 1996; Cazetta et al.,
2007). In Nigeria, corn is processed to a variety of diets including pap which is a major diet used
for weaning, and the capacity for corn production in Nigeria is high. Corn cobs form about 30 %
of maize agro-wastes of which application in bioethanol industry are the focus of many
researches aimed at achieving an effective and efficient waste management scheme (Olsson
and Hahn-Hägerdal, 1996).
Bio-ethanol can be produced by using different technologies. One of the most important
technology, fermentation, produces bio-ethanol by means of biological transformation of natural
starch and sugars resources such as energy-rich crops, (first-generation biofuels) and
lignocellulosic biomass (second-generation biofuels).
Most current bioethanol production processes (1st generation) utilize more easily degradable
biomass feedstock’s such as cereals (corn or grain) and sugarcane juice. However, the utilization
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of these agricultural crops exclusively for energy production is heavily conflicting with food and
feed production (Ikechukwu, 2012). Great effort is enforced on advancing a cellulosic bioethanol
concept (2nd generation) that utilizers lignocellulosic biomass.
Ethiopia, one of the world’s centers of genetic diversity in crop germplasm produces more of
maize than any other crop (CSA 2010). The area under maize cultivation in 2009/2010 was 1.69
hectares from which 37.8 million quintals of maize were produced which was higher than that of
any other cereal crop (Geta, 2013).
Corncob, a waste product of corn contains large amount of sugars that can be further utilized to
produce various compounds (Yah et al., 2010). The bioconversion of lignocellulosic to biofuel
from cheap non-edible materials such as corncob for renewal energy is imperative. Corncobs
contains sufficient amount of cellulosic material, which is the best source of fermentable sugars.
Corncob consists of polymers of mainly two types of sugars: glucose and xylose (Hsu, 2008).
The use of cobs in cellulosic ethanol production creates an identical alternative to grain produced
ethanol and reduces dependence on corn grain (Zych, 2008). Agricultural residue is gaining
much importance in these days because of its abundance, low cost, whole year decentralized
availability for the biological production of industrial chemicals such as glucose, furfural,
hydroxymethyl furfural, levulenic acid, lactic acid, acetic acid, propionic acid and fuels.
1.2 Statement of the Problem
In recently, due to the environmental concerns about air pollution caused by the combustion of
fossil fuels, thus an alternative energy sources need to be renewable, sustainable, efficient, cost-
effective, convenient and safe. The use of food crops (like corn, maize) for biofuels production
may cause inflation of cost of these crops leading to food insecurity. To alleviate such problems,
alternative and non-edible agricultural products must be investigated. One of the motivations for
developing biofuels is to lower carbon emissions. Thus, corncob can be used for the production
of second generation biofuel. In addition to environment benefit, bioethanol production from
corncob can stimulate community based jobs and economic growth. This project aims on
production of the bioethanol from corn cob which fills the energy gap and environmentally
friendly alternative source of energy.
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1.3 Objective
1.3.1 General objective
The general objective of this study is production of bioethanol from corn cob.
1.3.2 Specific objective
The specific objectives of the project are the following;

 To know the different methods of production process of bioethanol.
 To know the compositions of corn cob.
 To know the different types of bioethanol feedstock.
 To compute material and energy balance.
 To size the major equipment’s that are involved during the production process.
 To investigate the factors affecting yield of bioethanol.
1.4 Significance of the project
This project has great significance in terms of assuring the production of an alternative form of
energy from corncobs; which is locally available, and abundant. This project also highly
contributes in the substitute fossil fuel by biofuel. Fossil fuels are quickly being depleted due to
extensive and continuing over-utilization. If consumption goes in this rate the fossil fuel reserve
will be depleted completely within short period of time. In addition to this, continuous burning of
fossil fuel increases emission of greenhouse gasses to the atmosphere and causes global
warming. As , a renewable and non-food competitive feedstock raw material is desirable for the
production of alternative fuel oil such as bioethanol; corncob is one of such renewable and non-
food competitive raw material.

2 LITERATURE REVIEW
2.1 Background
Bioethanol is biofuel made through the fermentation of plant sugars from agricultural crops and
biomass resources (NEVC, 1998). With rapid depletion of the world reserves of petroleum,
bioethanol in recent years has emerged as one of the alternative liquid fuel and has generated
immense activities of research in the production of ethanol and its environmental impact.
Production of alcoholic beverages is in fact as old as human civilization. The production of pure
ethanol apparently begins in the 12-14th century along with improvement of distillation. During
the middle ages, alcohol was used mainly for production of medical drugs but also for the
manufacture of painting pigments. The knowledge of using starchy materials for ethanol
production was first employed in the 12th century in typical beer countries like Ireland.
Due to the tax imposition on bioethanol to assist in financing the civil war and the cheaper price
of kerosene, it quickly replaced bioethanol in 1861 (Morris, 1993). It was only in the 19th
century that this trade became an industry with enormous production figures due to the economic
improvements of the distilling process. It was at the beginning of the20th century that it had
become known that alcohol might be used as fuel for various combustion engines, especially for
automobiles. In the 1970‟s, the interest in fuel ethanol was renewed due to the oil crisis.
Nearly 25 federal agencies administered various ethanol programs and the National Alcohol
Fuels Commission was established to study the potential for alcohol based fuels (Lansing, 1983).
Ethanol gained further support in 1980 when Chrysler, Ford and General Motors released
statements that ethanol with blends of up to 10% would be covered in their vehicle warranties
(RFA, 1998). Its market grew from less than a billion liters in 1975 to more than 39 billion liters
in 2006 and is expected to reach 100 billion liters in 2015 (Licht, 2006). Interest in the use of
biofuels worldwide has grown strongly in recent years due to the limited oil reserves, concerns
about climate change from greenhouse gas emissions and the desire to promote domestic rural
economies.
2.1.1 Overview of maize production in Ethiopia
Ethiopia is among the major maize producers in Sub Saharan African countries, where small
holder farmers dominate the major share of production. Maize, which is originated from South

America, is first introduced in Ethiopia in the 16th to 17th Century (ECEA, 2009). It is classified
as one of ’’warm weather cereal crop” and widely cultivated at altitudes ranging from 1500 -
2200 meters above sea level of Western, Southwestern, and Southern parts of the country.
According to the data obtained from CIMMYT, Ethiopia is the third largest producer of maize in
Eastern and Southern Africa, following South Africa and Tanzania. It accounts for about 10% of
the area and 12% of the production of the region. Maize yield levels are also slightly above the
regional average-about 1.7 metric tons/ha compared to 1.5 metric tons/ha for the whole region.
In fact, yield of maize is the second highest following South Africa, which is about 2.3 metric
tons/ ha (CIMMYT, 1999/2000).
In Ethiopia small-scale subsistence farmers, private commercial farmers and state farms grow
maize. Small holders plant maize mainly as a subsistence crop while the large modern farms
mainly produce for the market. According to CSA data, the average area planted for maize
during 1997/98-2001/02 was about 1,497,300 hectares and during 2003/04 - 2007/08 was
1,549,613 hectare. The share of the smallholder sector was about 95% of total maize production.
Maize is widely grown in Ethiopia; only three regional states contribute to 94% of the total
annual production. These regions are Oromia, Amhara and SNNP. According to a five years
(2003/04 - 2007/08) CSA data, the share of Oromia region was on the average, 60% of the total
Maize production in the country. This was followed by Amhara with 21.67% and SNNP with
12.55%. Thus the trend of the National maize production was totally dependent on the
production field of the three regions. Accordingly, 16 zones from Oromia, 5 zones from Amhara
and 7 zones from SNNP region are found to be producers of more than 100,000 quintals per year
in all the years from 2003/04 - 2007/08. In Oromia region, 11 of the 16 zones on average
produce more than 1,000,000 quintals annually. The major maize producing zones of Ethiopia
and their relative share of the national maize production is shown table 2.1 below (ECEA, 2009).

Table 2.1 Major Maize Producing Administrative Zones of Ethiopia
Zone Average production (in quintals)

West Gojam 3,209,274
Jimma 2,128,619
West Welega 1,795,239
East Welega 1,790,953
South Gonder 1,561,297
Arsi 1,460,934
Source: (CSA, 2003/04 – 2007/08)
2.2 Bioethanol and its characteristics
Bioethanol or fuel alcohol refers to ethyl alcohol produced by microbial fermentation (as
opposed to petro chemically-derived alcohol) that is used as a transportation biofuel. It is
produced through distillation of the ethanol wash emanating from fermentation of biomass
derived sugars and can be utilized as a liquid fuel in internal combustion engines, either neat or
in petrol blends (Walker, 2011). Table 2.1 summarizes some of the important characteristics of
bioethanol as a fuel source.
Table 2.2 Physico-chemical characteristics of bioethanol
No. parameter Characteristic properties

1 Molecular formula C2H5OH
2 Molecular mass 46.07 g/mol.
3 Appearance Colorless liquid
4 Water solubility (between –117°C and 78°C)
5 Density 0.789 kg/l
6 Boiling temperature 78.5°C (173°F)
7 Freezing point –117°C
8 Ignition temperature 425°C
9 Acidity (pKa) 15.9
Source: (Walker, 2011)

2.3 Bioethanol feedstock’s
There is a growing interest worldwide to find out new and cheap carbohydrate sources for
production of bioethanol (Mohanty et al., 2009). For a given production line, the comparison of
the feedstock’s includes several issues (Gnansounou et al., 2005) (1) chemical composition of
the biomass (2) cultivation practices (3) availability of land and land use practices (4) use of
resources (5) energy balance (6) emission of greenhouse gases, acidifying gases and ozone
depletion gases (7) absorption of minerals to water and soil (8) injection of pesticides (9) soil
erosion (10) contribution to biodiversity and landscape value losses (11) farm-gate price of the
biomass (12) logistic cost (transport and storage of the biomass) (13) direct economic value of
the feedstocks taking into account the co-products (14) creation or maintenance of employment
and (15) water requirements and water availability. Bioethanol feedstocks can be divided into
three major groups: (1) First generation feedstocks, (2) Second generation feedstocks and (3)
third generation feedstocks.
2.3.1 First generation feedstocks
First generation bioethanol feedstocks come from agricultural cereal and sugar crops that are also
sources of human (and animal) food. The bioethanol produced by fermentation of sugars such as
sugarcane (Macedo et al., 2008; Leite et al., 2009), sugar beet (Ogbonna etal. 2001; Icoz et al.,
2009), sorghum (Mamma et al., 1995; Prasad et al., 2007a; Yu et al.,2008), whey (Domingues et
al., 2001; Gnansounou et al., 2005; Silveira et al., 2005; Dragoneet al., 2009) and molasses
(Roukas, 1996) and starchy feedstocks such as grains viz. maize(Gaspar et al., 2007; Persson et
al., 2009), wheat (Nigam, 2001), root crops such as cassava(Amutha and Gunasekaran, 2001;
Kosugi et al., 2009; Rattanachomsri et al., 2009) are commonly known as first generation
bioethanol.
Sugar crops need only a milling process for the extraction of sugars to fermentation (not
requiring any step of hydrolysis), becoming a relatively simple process of sugar transformation
into ethanol. In this process, ethanol can be fermented directly from cane juice or beet juice or
from molasses generally obtained as a byproduct after the extraction of sugar (Icoz et al., 2009).
In processes that use starch from grains like corn, saccharification is necessary before
fermentation. In this step, starch is gelatinized by cooking and submitted to enzymatic hydrolysis

to form glucose monomers, which can be fermented by microorganisms (Mussattoet al., 2010).
First generation bioethanol have played an important role in establishing the infrastructure and
policy drivers, required to support renewable transport fuels in the international market place
(EIA, 2008). There are examples of various first generation crops having various amount of
ethanol production. However its growth and development is limited due to (i) competition with
food and fibre production for the use of arable land (ii) regionally constrained market structures
(iii) lack of well managed agricultural practices in emerging economies (iv) high water and
fertilizer requirements and (v) a need for conservation of bio-diversity.
2.3.2 Second generation feedstocks
Exploitation of first generation feedstocks for future bio-fuel production is ultimately

unsustainable due to food security and land-use issues. Second-generation bioethanol refers to
fuel alcohol produced from non-food biomass sources, such as lignocellulose, the most abundant
form of carbon on the earth. The various forms of lignocellulosic feedstocks can be grouped into
six main categories.
They account for nearly 50% of world biomass with an estimated annual production of 10 to 50
billion tons, making lignocellulose arguably the most abundant and renewable organic
component of the biosphere (Claassen et al., 1999). Lignocellulosic biomass in the form of wood
and agricultural residues is virtually inexhaustible, since their production is based on the
photosynthetic process which is about 60% of the total biomass produced (Kuhad et al., 1997). It
was estimated that terrestrial plants produce about 1.3×1010 metric tons per annum which is
energetically equivalent to about two-thirds of the world‟s energy requirement (Kim and Yun,
2006).
Moreover, agricultural residuals or by-products are annually renewable, abundantly available and
account for more than 180 million tons per year (Kapdan and Kargi, 2006). These lignocellulosic
biomass includes woody material (Ballesteros et al., 2004), straws (Huang et al., 2009; Silva et
al., 2010), agricultural waste (Lin and Tanaka, 2006) and crop residues (Hahn-Hagerdal et al.,
2006). Second-generation biofuels are expected to reduce net carbon emission, increase energy
efficiency and reduce energy dependency, potentially overcoming the limitations of first-
generation biofuels (Antizar-Ladislao and Turrion-Gomez, 2008).

The other major benefits of switching to cellulosic ethanol are its renewable nature, long term
sustainability, low net carbon emission, high energy efficiency, low energy dependency, increase
in national security and diversifying rural economies (IEA,2008b). Polysaccharides present in
lignocellulosic materials including cellulose and hemicellulose are of great interest as feedstocks
for second generation bioethanol production.
Lignocellulosic biomass
Lignocelluloses are complex substrates composed of a mixture of carbohydrate polymers i.e.
cellulose and hemicelluloses tightly bound to lignin, a complex aromatic polymer, mainly by
hydrogen bonds but also by some covalent bonds. Lignin interferes with cellulose hydrolysis
because it acts as a physical barrier that prevents the contact of cellulase to cellulose. The first
step in biofuels production from lignocelluloses therefore is delignification to liberate cellulose
and hemicelluloses from their complex with lignin. It is a very crucial, rate limiting and difficult
task (Kudirat, 2012).
The chemical compositions (cellulose, hemicellulose and lignin content) of various feedstocks
for bioethanol production on average, agricultural wastes with the highest cellulose content are
corn cob, softwood stems, cotton seed hairs and saw dust. Forest residues comprise about 80% of
the world’s biomass (Demirbas, 2005) and in the United States alone 33.5-44.6 million metric
tons of corn cob are available for harvest each year (Zych, 2008).
Composition of Lignocellulosic biomass
Lignocellulosic materials do not contain monosaccharide’s that are readily available for
bioconversion but polysaccharides such as cellulose and hemicellulose, lignin, extractives, and
ashes. The polysaccharides need to be hydrolysed by means of either enzymes or acids to
fermentable sugars (Talebnia, 2008).
Figure 2.1 Lignocellulose structure, showing cellulose, hemicellulose and lignin fractions
A. Cellulose

Cellulose is an unbranched homo polysaccharide composed of β-D glucose units linked by (1,4)
glycosidic bonds. However, the basic building block of cellulose is a dimer of two glucose units
known as cellobiose (See Figure. 2.3). Cellulose is the most abundant material on Earth, and it is
the main constituent of plants. It is also present in bacteria, fungi, algae and even in animals. In
nature, cellulose chains have a degree of polymerization (DP) of approximately 10,000 and
15,000 glucopyranose units in wood and native cotton celluloses, respectively (Talebnia, 2008).
Figure 2.2 Schematic illustration of the cellulose chain
B. Hemicellulose
Hemicellulose or polyose is a mixture of polymers comprising pentoses, hexoses hexuronic acids

and deoxy-hexoses. Hemicelluloses differ from celluloses by composition of various sugar units
and by much shorter and branched molecular chains. In contrast to cellulose which is crystalline,
strong, and resistant to hydrolysis, hemicellulose has a random, amorphous structure with little
strength. Therefore, it is easily hydrolyzed by dilute acid or base, as well as hemicellulose
enzymes (Talebnia, 2008).
C. Lignin
Lignin is a complex, hydrophobic, cross-linked, three-dimensional aromatic polymer of phenyl

propane building blocks. The mechanical strength properties of plants are mainly due to
incorporation of lignin into their cell walls, whereby huge plants such as trees can remain
upright. Lignin is one of the most complicated natural polymers with respect to its structure and
heterogeneity, which make it extremely resistant to chemical and biological degradation
(Talebnia, 2008).

2.3.3 Third generation feedstocks
Third-generation biofuels are produce from algal biomass, which has a very distinctive growth
yield as compared with classical lignocellulosic biomass (Brennan and Owende,2010).
Microalgae have broad bioenergy potential as they can be used to produce liquid transportation
and heating fuels, such as biodiesel and bioethanol. Microalgae provide carbohydrates (in the
form of glucose, starch and other polysaccharides), proteins and lipids for the production of
biofuels.
2.4 Corncobs as a Feedstock of Bioethanol
The maize plant comprise of the stalks, husks, shanks, silks, leaf blades, leaf sheaths, tassels and
cobs. The corn cob carries the grain and together with associating husks, shanks and silks are
harvested from the farm. The other parts are left on the farm to rot. Corn cobs form about 30% of
maize agro-wastes (Kudirat, 2012).The agricultural residues from maize production are potential
sources of sugar for bioethanol production.
Before its use as a substrate for fermentation processes, the raw material has to be pretreated.
Pretreatment is one of the many steps in the cellulose-to-ethanol process, but represents a
currently critical step for hydrolysis. An effective pretreatment is performed at conditions that
avoid degradation of pentose from hemicellulose, or glucose from cellulose, and limit formation
of degradation products that inhibit the growth of fermentative microorganisms.
Corncobs are a lignocellulosic material composed of cellulose, hemicellulose and lignin. These
polymeric fibres consist of monomeric molecules. Cellulose is built of C6 sugars; hemicellulose
mainly of the C5 sugars xylose and arabinose. Lignin consists of phenolic macromolecules. Corn
cobs contain 32.3-45.6% cellulose, 39.8% hemicelluloses – mostly composed of pentoses and
6.7-13.9% lignin (Kudirat, 2012; Sun and Cheng, 2002) and can be converted to fermentable
sugar for ethanol production.
The cobs produced from corn are underutilized, being mostly used as manure for agricultural
production or burnt as fuel in households (Yah et al., 2010). Utilization of corncobs for second
generation bioethanol production has been reported in several studies and represents a valid
opportunity for the utilization of such feedstock.

2.5 Bioethanol Production Process
Ethanol can be produced in two different ways. Either chemically, by hydration of ethylene,
which is derived from crude oil or natural gas, or by fermentation of sugar containing feeds,
starchy feed materials or lignocellulosic materials. About 5% - 10% of the ethanol produced in
the world is a petroleum product. Petroleum ethanol product is made by the catalytic hydration of
ethylene with sulfuric acid as the catalyst. It can also be obtained via ethylene or acetylene, from
calcium carbide, coal, oil gas, and other sources.
The two primary ways of producing bioethanol from cellulosic feedstock are:
 Biochemical conversion process.
 Thermo chemical conversion process.
2.5.1 Biochemical Conversion
Biological conversion consists of exposing biomass to certain microorganisms. The secondary

fuels produced are the result of metabolic activity of the microorganisms. Production of ethanol
through fermentation through anaerobic digestion is the most common biological conversion
processes. Ethanol fermentation from carbohydrates is probably one of the oldest processes
known to man. Today, it is widely regarded as an important potential alternative source of liquid
fuels for the transport sector (NUEP, 2013).
Typical lignocellulose-to-ethanol processes consist of at least four steps. These are pretreatment
to enhance biomass digestibility, hydrolysis of cellulose to sugar monomers, fermentation of
sugars to ethanol, and recovery of ethanol by distillation/evaporation from process stream
(Ayele, 2011).
A. Pretreatment
Pretreatment is required to alter the biomass macroscopic and microscopic size and structure as
well as its submicroscopic chemical composition and structure so that hydrolysis of carbohydrate
fraction to monomeric sugars can be achieved more rapidly and with greater yields (Sun and
Cheng, 2002;Ikechukwu, 2012).
The effect of pretreatment of lignocellulosic materials has been recognized for a long time. The
purpose of the pretreatment is to remove lignin, reduce cellulose crystalline and increase the
porosity of the materials. Pretreatment must meet the following requirements: Improve the

formation of sugars or the ability to subsequently form sugars by acidic or enzymatic hydrolysis;
avoid the degradation or loss of carbohydrate; avoid the formation of byproducts inhibitory to the
subsequent hydrolysis and fermentation processes.
Physical, chemical, Physico-chemical, and biological processes have been used for pretreatment
of lignocellulosic materials (Sun, Y. and Cheng, J. 2002).The purpose of the pretreatment was to
reduce cellulose crystallinity and increase the porosity of the materials. Pretreatment must meet
the following requirements: improve the formation of sugar, avoid the degradation or loss of
carbohydrate, avoid the formation of by-product inhibitors and must be cost effective.
Dilute acid pretreatment
Acid pretreatment firstly developed in Germany in 1898. In this method concentrated or dilute
mineral acids like sulfuric acid are used in order to break down hemicelluloses into monomeric
sugars and simultaneously removing part of the lignin. This method needs a small amount of
water since a small amount of energy is required to get an optimum temperature (Dehnavi,
2009). Dilute acid hydrolysis is the most employed technique for the hemicellulose breakdown.
In this process, the use of diluted acids (1 to 4%) under moderate temperatures (120 to 160°C)
has proven to be adequate for hemicellulose hydrolysis while promoting little sugar
decomposition. (Mussatto, 2010: Vincent, 2010).
Another study carried by (Bensah, et al., 2013) showed that under dilute acid (0.2–2.5% w/w)
processes, high temperatures (120–210°C) and pressures are used to achieve reaction times in
seconds or minutes and are thus suitable for continuous operations. The low acid consumption is
a major advantage in terms of cost and process severity. Moreover, low acid concentrations
(<1% w/v sulphuric/phosphoric) release essential nutrients (S and P) that enhance downstream
fermentation.
Advantages
 High yield of hemicelluloses sugar,

 Remove of lignin and hemicelluloses in this method increases exposing of cellulose to
enzyme,
 Remove of heavy metals in the raw materials (Dehnavi, 2009).

B. Hydrolysis
Hydrolysis is the unit operation that depolymerizes the polysaccharide chains of cellulose and
hemicellulose into fermentable oligosaccharides and/or monosaccharide’s (Alicia, 2013). After
the pretreatment process, there are two types of processes to hydrolyze the feedstocks for
fermentation into ethanol, most commonly used are acid (dilute and concentrated) and enzymatic
hydrolysis. In addition, there are some other hydrolysis methods in which no chemicals or
enzymes are applied. For instance, lignocellulose may be hydrolyzed by gamma-ray or electron-
beam irradiation, or microwave irradiation. However, those processes are commercially
unimportant.
Hydrolysis of cellulosic materials includes the processing steps that convert the carbohydrate
polymers e.g. cellulose and hemicellulose into monomeric sugars. Cleavage of these polymers
can be catalyzed enzymatically by cellulases or chemically by acids such as sulfuric acid (Mosier
et al., 2005). The factors that have been identified to affect the hydrolysis of cellulosic biomass
include porosity or accessible surface area, cellulose fiber crystallinity, and the content of lignin
and hemicellulose (Talebnia, 2008). Both enzymatic and chemical hydrolyses require a
pretreatment to increase the susceptibility of cellulosic materials (Demirbas, 2005).
Dilute Acid Hydrolysis
The dilute acid process is conducted under high temperature and pressure, and has a reaction
time in the range of seconds or minutes, which facilitates continuous processing. The
combination of acid and high temperature and pressure dictate special reactor materials, which
can make the reactor expensive. The first reaction converts the cellulosic materials to sugar and
the second reaction converts the sugars to other chemicals. Unfortunately, the conditions that
cause the first reaction to occur also are the right conditions for the second to occur (Demirbas,
2005).
The principle of acid hydrolysis is to apply temperature and pressure in order to soften
lignocellulosic providing better penetration of the acid, and then degrade carbohydrate part of
wood into monosaccharide’s. During treatment various products are formed: monosaccharide’s
(xylose, arabinose, mannose etc.), some sugar-dehydration products (furfural, hydroxyl methyl
furfural), while lignin and part of cellulose remain as solid residue. Research works on the dilute
acid hydrolysis of different lignocellulosic materials have defined optimal process conditions:

temperature 80-200°C, sulfuric acid concentration 0.25–8 wt. %, and reaction time 10-2000 min.
Sulfuric acid is a commonly used acid due to low cost, non- volatilizes and affordable corrosion
strength (Gladyshko, 2011).
The biggest advantage of dilute acid processes is their fast rate of reaction, which facilitates
continuous processing. Since 5-carbon sugars degrade more rapidly than 6-carbon sugars, one
way to decrease sugar degradation is to have a two-stage process. The first stage is conducted
under mild conditions to recover the 5-carbon sugars while the second stage is conducted under
harsher conditions to recover the 6-carbon sugars (Demirbas, 2005).
C. Fermentation
Lignocellulose is often hydrolyzed by acid treatment. The hydrolysate obtained is then used for
bioethanol fermentation by microorganisms such as yeast. Because such lignocellulose
hydrolysate contains not only glucose, but also various monosaccharide’s, such as xylose,
mannose, galactose, arabinose, and oligosaccharides, microorganisms should be required to
efficiently ferment these sugars for the successful industrial production of bioethanol.
The fermentation of ethanol is the biological process that converts fermentable sugars such as
glucose and xylose to cellular energy with microorganisms which produce waste by-products
ethanol and carbon dioxide anaerobically by the metabolic pathways of sugars. This is unlike
yeast cells S. cerevisiae (baker‟s yeast), Schizo saccharomyces pombe be that prefer
fermentation even in the presence of oxygen and will produce ethanol given a suitable source of
nutrition. S. cerevisiae is yeast that is most widely used in the production of ethanol from
hexoses but cannot utilise pentoses.
In general, the conversion of lignocellulosic material to sugar and then ethanol is governed by
equation below:
(C6H10O5) n + nH2O nC6H12O6 + yeast 2nC2H5OH + 2nCO2 (2.1)
From the above equation the first step is hydrolysis and the second step is fermentation.
According to the reactions, the theoretical maximum yield is 0.51 kg bioethanol and 0.49 kg
carbon dioxide per kg of xylose and glucose. The overall reaction of this fermentation of hexose
sugar (glucose) by yeast has been expressed by Gay-Lussac which forms the basis of calculating
fermentation efficiency as:
3C5H10O5 5C2H5OH + 5CO2……………………………………………………...... (2.2)
C6H12O6 2C2H5OH + 2CO2 ……………………………………………………….... (2.3)

Saccharomyces cerevisiae is the most favored organism for ethanol production from hexoses. P.
stipites and Candidashehatae are capable of fermenting both hexose (glucose) and pentose
(xylose) sugars to ethanol (Joshi, 2011).
Xylose-fermenting microorganisms are found among bacteria, yeast and filamentous fungi.
Today, xylose fermenting bacteria include both native and genetically engineered organisms, and
many have characteristics useful for simultaneous saccharification and fermentation (Ayele,
2011).
One of the most effective bioethanol producing yeasts, S. cerevisiae, has several advantages
owing to its high bioethanol production from hexoses, high tolerance to bioethanol and other
inhibitory compounds in the acid hydrolysates of lignocellulosic biomass. However, because
wild-type strains of this yeast cannot utilize pentoses, such as xylose and arabinose bioethanol
production from a lignocellulose hydrolysate is inadequate. For xylose-using S.cerevisiae, high
bioethanol yields from xylose also require metabolic engineering strategies to enhance the xylose
(Ayele, 2011).
The optimal pH range for Saccharomyces cerevisiae varies between 4.0 and 6.0 depending upon
the fermentation medium. The pH affects the efficiency of ethanol fermentation by influencing
the activity of plasma proteins and intracellular enzymes. If enzymes are deactivated by pH < 4.0
the yeast will not be able to grow and produce ethanol efficiently (Uncu, 2009) observed an
increase in ethanol production as well as fermentation efficiency with an increase in pH from 4.0
to 5.0 and found the optimum pH for Saccharomyces cerevisiae species around pH 4.5. Another
study carried by (Yalçın and Özbaş, 2008 as cited in Uncu, 2009) showed that Saccharomyces
cerevisiae worked well between a pH range of 4.0-4.5 and yielded slightly better results at pH
4.0 with fermentation of Kalecik Karası and Narince types of grapes.
One of the most successful microorganisms for bioethanol production is Saccharomyces

cerevisiae. Although the wild-type strain has high bioethanol productivity and very tolerant to
high ethanol concentrations and inhibitory compounds, it is unable to ferment pentoses
(hemicelluloses). According to study (Sumphanwanich al, 2008) reported that 0.45 g ethanol/g
glucose. Pichia stipitis, Candida shekhatae and Pachysolan tannophilus are promising microbes

that are capable of fermenting both hexoses and pentoses. However, S. cerevisiae is still the most
commercialized and dominated strains for bioethanol production (Harun et al., 2009).
Microorganisms for bioethanol fermentation can best be described in terms of their performance
parameters and other requirements such as compatibility with existing products, processes and
equipment. The performance parameters of fermentation are temperature range, pH range,
alcohol tolerance, growth rate, productivity, osmotic tolerance, specificity, yield, genetic
stability, and inhibitor tolerance (Demirbas, 2004). All the recombinant strains are mesophilic
organisms and function best between 25℃, and 33℃.
C. Separation
Distillation
Distillation is one of the steps of the purifications. Distillation is the method used to separate two
liquid based on their different boiling points. However, to achieve high purification, several
distillations are required. This is because all materials have intermolecular interactions with each
other, and two materials will co-distill during distillation. (Onuki, 2005 as cited in Wondale,
2012).
Dehydration
After distillation, some amount of water remains in bioethanol. Especially, this water is a big
problem for fuel ethanol because the presence of this amount of water enhances the molecular
polarity of bioethanol when it is mixed with gasoline. Consequently, they separate into two
phases, ethanol phase and gasoline phase. It is easy to imagine that this inhomogeneous fuel is
not acceptable. Thus, dehydration can be another issue (Onuki, 2005).
For the bioethanol to be usable as a fuel, water must be removed. Most of the water is removed
by distillation, but the purity is limited to 95-96% due to the formation of a low boiling water-
ethanol azeotrope. For blending with gasoline, purity of 99.5 to 99.9% is required, depending on
temperature, to avoid separation. Currently, the most widely used purification method is a
physical absorption process using molecular sieves. Another method is azeotropic distillation
(Onuki, 2005as cited in Wondale, 2012).

2.5.2 Thermochemical Process
There are two bioethanol production processes that currently employ thermochemical reactions
in their processes. The first system is actually a hybrid thermochemical and biological system.
Biomass materials are first thermo chemically gasified and the synthesis gas (a mixture of
hydrogen and carbon oxide) bubbled through specially designed fermenter. A micro-organism
that is capable of converting the synthesis gas is introduced into the fermenters under specific
process condition to cause fermentation to bioethanol.
The second thermochemical bioethanol production process does not use any micro-organisms. In
this process, biomass materials are first thermo-chemically gasified and the synthesis gas passes
through a reactor containing catalysts, which cause the gas to be converted into ethanol. An
intensive effort was made to develop these processes for fuel. Numerous efforts have been made
since then to develop commercially viable thermochemical-to-ethanol processes.
Bioethanol yields up to 50% have been obtained using synthesis gas-to-ethanol processes. Some
processes that first produce methanol and then use catalytic shifts to produce ethanol have
obtained ethanol yields in the range of 80%. Unfortunately, like the other processes, finding a
cost-effective all-thermochemical process has been difficult. (Badger, P.C. 2002).
Advantages of cellulosic bioethanol
The advantages of cellulosic ethanol over starch ethanol are as follow:
 According to Regmi et al (2001), a 1% increase in food prices causes and average 0.75%
decline in food consumption in developing countries. In addition to reducing caloric
intake as food prices increase, low-income people also switch to less nutritious food by
Von Braun (2007).
 Cellulosic bioethanol as a viable alternative for reducing oil dependence while protecting
crops i.e. way to prevent the displacement of crops that feed humans.
 The fuel is produced from agriculture byproducts; it has no impact on the food supply or
land use.
 Cellulosic bioethanol could help reduce air pollution- cellulosic ethanol not only emits
less greenhouse gas than gasoline but emits fewer fine particles into the air.
 From the analysis carried out, it is apparent that a shorter residence time is required for
the production of bioethanol from cellulose.

 It provides a much greater energy yield.

Disadvantages of cellulosic bioethanol
 It involves more complex methods of production.
 It is very expensive to produce.
Extra uses of Bioethanol
The following are some of the various uses of bioethanol in addition to the fuel:
1. Raw materials: it is a raw material in the manufacturing of acetaldehyde, acetic acid,
ethylene, glycol, dyes, detergents and cleaning solution.
2. Pharmaceutical preparation: bioethanol is used in the preparation of cough syrups. It is also
used in the preparation of antiseptics.
3. Miscellaneous: It is used as anti-freeze in automobile radiator. It is also used in preparation of
alcoholic beverages.
2.6 Factors Affecting Fermentation
Microorganisms for bioethanol fermentation can best be described in terms of their performance
parameters and other requirements such as compatibility with existing products, processes and
equipment. The performance parameters of fermentation are temperature, pH, alcohol tolerance,
growth rate, productivity, specificity, yield, genetic stability, and inhibitor tolerance (Demirbas,
2005).
A. Effect of sugar concentration
The concentration of sugar can affect the microbial bioethanol fermentation in various ways. Use
of concentrated sugar substrate is one of the ways to obtain high bioethanol yield during
fermentation. The amount of bioethanol produced is proportional to the amount of sugar added;
thus, high sugar concentrations are desired. However, too high sugar concentrations can inhibit
metabolism due to increased osmotic pressure. Very low levels of sugar may limit the rate of
ethanol production (Jones et al., 1981).
B. Effect of temperature
Temperature has an important factor on the growth rate of the microorganisms and the rate of
bioethanol production. Wine and beer fermentations are generally conducted below 20°C, 27
whereas higher temperatures (30-38°C) are being examined for industrial alcohol production by
yeast cultures (Sofer and Zaborsky, 1981). Too high temperature kills yeast, and low temperature
slows down yeast activity and growth. Thus, specific range of temperature is required (Onuki,

2005). The enzyme hydrolysis process for saccharification able to operate up to 55 °C may be
combined with fermentation, further reducing capital and glucose inhibition (Hettenhaus, 1998).
C. Effect of PH
A very important factor for cellular growth is external PH. Most alcoholic yeast fermentations
are conducted below pH 4.5, although this may not be the optimal pH for growth or ethanol
production. Yeast cultures can grow over a wide range from 3 to 8 with an optimum for growth
generally in the slight acidic range. Shifts in pH can also affect the final ratio of organic waste
products produced by yeast cultures. Thus, the optimal pH for a fermentation process must
support a balance among ethanol production, cellular growth, and physicochemical effect on
waste product pathways. Low pH values in yeast fermentation help to inhibit growth of
contaminating bacterial cultures. Bacterial cultures generally have a pH optimum around 7-7.5,
with less tolerance than yeast to acid conditions (Sofer and Zaborsky, 1981).

3 METHODOLOGY
3.1 Materials
The materials used to run all experiments were listed below:

Chemicals:
 Sodium Hydroxide (NaOH)
 Sulphuric acid (H2SO4)
 Dextrose sugar
 Yeast extract
 Yeast (Saccharomyces cerevisiae).
Equipments:
 Digital balances
 Vacuum Filter
 Sieves (mesh size of 2.0 mm)
 Shaking Incubator
 Autoclave
 pH- Meter
 heating mantle
 Condenser
 Vessels
 Fourier Transform Infrared spectroscopy (FTIR).
3.1.1 Characterization of corncob
To determine the moisture content, fixed carbon content, ash content and volatile matter content
of air-dried biomass samples ground to particle size below 2.0 mm.
Moisture Content
Samples were weighed in clean preheated moisture crucible of known weight by using sensitive
balance. The sample and crucible were kept in an oven 105℃ for an hour. The crucible was
covered and transferred to desiccators, and weighed after reaching room temperature. The
crucible was heated in the oven for another two hours and was re-weighed. This was repeated
until constant weight was obtained. The loss of weight was calculated as percent of weight and
expressed as moisture content.

Moisture content (%) = *100
Where: W1= Initial weight, W2= weight after drying

Volatile Matter Content
A crucible was weighed empty, and then samples were put in it. The sample and the crucible
were placed in a muffle furnace for 30min at 600oC. The crucible was removed from furnace and
placed in a desiccators to cool, then was reweighed. The process was repeated until constant
weight was obtained.
Volatile content (%) =
Where: W1= Original weight of the sample, W2 = Weight of sample after cooling
Ash Content
A crucible was weighed empty, and then samples were put in it. The sample and the crucible
were placed in a muffle furnace for 2 hours at 550℃. The crucible was removed from furnace
and placed in a desiccators to cool, then was reweighed
Ash content (%) = =

Where: W1= Original weight of the sample, W2 = Weight of sample after cooling
Fixed Carbon Content
This is the residue left after the moisture, volatile and ash is given up. It is deduced by
subtracting from 100, the percentage of moisture, volatile matter and ash content. The fixed
carbon content (FC) is given as;
FC = 100 – (%moisture + %volatile matter + % ash)
Methods of production process
As we discussed in the literature review there are two methods of bioethanol production. From
these methods of production we select biochemical conversion due to the following reasons;
 Gives high yield,
 Reduces environmental pollution,
 Needs less operating energy.
The bioethanol is produced biochemically through the following steps.
3.2 Corn cob Preparation
First raw material is collected form the place where there is mass production of maize. Corn cob
is then rinsed in water, drained and sundried for specified days. The Cob is further treated by

breaking to small pieces with the aid of wooden mortar and pestle in such a way that it is suitable
to be dried and ground.
Then corncob is ground using milling machine to the size of 2mm and sieve analysis was
performed. Flour whole size was greater than 2mm was again ground. Grinding of Corncob into
fine powder gives increased surface area which enhances the contact between hemicellulose and
cellulose.
a) Corn cob b) grounded corn cob
Figure 3.1 corn cob and grounded corn cob
3.3 Acid pretreatment
Dilute acid hydrolysis pretreatment is used for biofuel production apply from 0.05 to 2% H2SO4
(w/v) at temperature range between 120 and 220℃. Corn cob powder is pretreated inside
autoclave and heated at temperature of 120℃, for 30 minutes. After that, it is cooled and filtered.
The filtrated is preserved in another vessel prepared for this purpose and kept it for fermentation.
The residue is washed twice by distilled water to remove sulfuric acid from it and kept for
hydrolysis purpose.
3.3 Dilute Acid Hydrolysis
Dilute acid hydrolysis is an easy and productive process. Research works on the dilute acid
hydrolysis of different lignocellulosic materials have defined optimal process. The acid
hydrolysis procedure started with adding of diluted sulfuric to the non-soluble component from
pretreatment steps and the corncobs were hydrolyzing in the reactor. Next separate the solid

particles from the liquid in the hydrolysate by vacuum filtration (to remove the non-fermentable
lignin portion). After separating the solid part, the solid part is washed with distilled water.
Finally, mixing the soluble component with the previously filter solution from the pretreatment
step for the next procedure.
PH Adjustment
Before addition of any micro-organism to the above prepared solution, pH of these samples has
to be adjusted. Otherwise the micro-organism will die in hyper acidic or basic state. A pH of
around 5.0 -5.5 is maintained. Pretreated and hydrolyzed solutions are mixed, shaken substrate
primarily checked for pH using a pH meter. Since, the mixed solution is more acidic media, and
then it would maintain the pH -5 adding sodium hydroxide solutions.
3.4 Fermentation
Microorganism used for fermentation
Baker’s yeast, Saccharomyces cerevisae used for fermentation is cultured on yeast extract agar.
In order to prepare the media should have the favorable condition for yeast growth or to supply
the required amount of nutrients. The following nutrients are mixed in their correct proportion.
 Dextrose,
 Malt yeast extract,
 Distilled water.
Sterilization: The reactor and all the equipment’s that are used for fermentation purposes are
sterilized (autoclaved). The sterilization is carried out at a specified temperature and time which
is condisive for the micro-organism culture.
Fermentation: The prepared solution and media are mixed in the fermenter with a specified
ratio. Then, it placed on shaking incubator for uniform temperature and composition.
3.5 Bioethanol Separation
The fermented product is distilled using distillation. The bioethanol is separated based on the
boiling point difference. Finally the bioethanol is harvested with small amount of water.

Corn cob
Pretreatment Hydrolysis
Preparation
Fermentation Distillation Molecular

Sieve Bioethanol
Yeast Bottom product Water
Figure 3.2 Production process of bioethanol from corn cob

4 RESULTS AND DISCUSSION
In this section the study discussed proximate analysis and chemical composition of the sample,
effect of acid hydrolysis on sugar and bioethanol yield.
4.1 Characterization of corncob
Proximate analysis
Table 4.1 Proximate analyses of the corncob sample
Physical composition Weight percentage (%wt. dry basis)

Moisture 6.8
Volatile 78.2
Ash 0.5
Fixed carbon 10
The sample of corncob with higher moisture content needs more heat for moisture vaporization.
Ash is a measure of inorganic impurities in the corncob. In this study low ash content of corncob
constituents, so decreasing sludge formation in the bioethanol production. Finally, fixed carbon
(FC) it is the carbon found in the material which is left after volatile materials are driven off this
is used for the determination of carbon in the corncob.
Chemical composition analysis
Table 4.2 chemical composition of corncob sample
Chemical composition Weight percentage (w/w %)

Cellulose 50
Hemicellulose 35.7
Lignin 12
Extractive 1.8
The determination of cellulose and hemicellulose can be applied to quantify the theoretical
production of bioethanol. However, Saccharomyces cerevisiae only converts glucose. In this
study, corncob contained high contents of the total cellulose of approximately 50% cellulose.
The lower the lignin content the easer hydrolysis condition, and decrease formation of toxic
chemicals such as, aromatic, polyaromatic, phenolic and aldehydic.

4.2 Effect of Acid hydrolysis on bioethanol yield
In this study, the total reduced sugar content through hydrolysis process was investigated. The
powdered corncob through hydrolysis at different acid concentration, hydrolysis time, and
temperature on the amount of sugar produced. Total reducing sugar of the hydrolyzate sample
was determined using calibration curve which was plotted from the known concentration of
standard glucose reacted with Benedict solution reagent and absorbance of standard glucose-
benedict solution after reaction.
Figure 4.1 Concentration of standard glucose and its absorbance

The effect of hydrolysis process variables

As shown from the figure 4.2 below the yield of TRS and bioethanol have direct relationship that
means, the run which has maximum sugar content has maximum yield of bioethanol. The same
is true in the run which has less sugar content also less yield of bioethanol. This situation
indicates the efficiency of fermentation process. The fermentation parameters are fixed for all
runs.
Figure 4.2 Total reducing sugar and the bioethanol yield from corn cob
Effect of time
The yield of bioethanol is slightly affected by hydrolysis time, as the hydrolysis time increase the
yield slightly increase. Beyond a certain hydrolysis time the yield of ethanol slightly decreases.

5 MATERIAL AND ENERGY BALANCE
5.1 Material and energy balance overview
Material and energy quantities, as they charge and discharge into and from process operations,
can be described by material and energy balances. Such balances are statements on the
conservation of mass and energy. If there is no accumulation, what goes into a process must
come out. This is true for continuous operation. If no input and output during the operation
carried out the process is batch process.
Material and energy balances are very important in an industry. Material balances are
fundamental to the control of processing, particularly in the control of yields of the products. The
first material balances are determined in the exploratory stages of a new process, improved
during pilot plant experiments when the process is being planned and tested, checked out when
the plant is commissioned and then refined and maintained as a control instrument as production
continues. When any changes occur in the process the material balances need to be determined
again.
The energy balance determinations are also made to determine the energy requirements of the
process, the heating, cooling and power required. In this plant operation it is thought that an
energy balance (energy audit) on the plant will show the pattern of energy usage and suggest
areas for conservation and savings.
5.2 Material Balance
Basis: 340 days per year.

Production rate: 5x106lit/year of ethanol (99.9%) with plant operation of 340 days per year. the
Plant attainment: The percentage of the available hours in a year.
% Plant attainment =
= = 93.15%.
The percent of plant attainment was acceptable because the recommended % of plant attainment
for chemical industry is between 90-95%.
The production rate in term of mass flow rate is given by;

ƍ =𝑚/v……………………………………………………………………………………… (5.1)
m = ƍ *V = 0.789 * 5x106
m= 3,945,000 or 3,945 ton/yr.

Production flow diagram of bioethanol from corn cob
Fresh corn cob water vapor
Heat Drying
H2SO4 Diluted acid pretreatment
Hydrolysis
H2O and H2SO4
Lignin
Filtration
NaOH
Neutralization
Inoculum
Fermentation
Bioethanol
Heat
Distillation Molecular sieve
Bottom product Water
Figure 5.1 Production flow diagram of bioethanol from corn cob

Since the annual production rate is known, so the material balance begins from the last unit
operation.

Material balance for Molecular sieve
Molecular sieve is an equipment which following the distillation column and bring the ethanol
concentration from 96% to 99.9%.
Anhydrous bioethanol (ṀAE) = 3945ton
Hydrous Bioethanol (ṀHE) =?
XHE = 96%
Xw = 4%
Water (W) =?
Xw=1, Xe = 0%
Figure 5.2 Material balance on molecular sieve

Where Xe = Percentage of Ethanol
Xw = Percentage of water
Over all mass balance:
ṀHE = W + ṀAE …………………………………………………………………………… (5.2)
Bioethanol component balance:
ṀHE* XHE = ṀAE* XAE…..……………………………………………………………… (5.3)
From equation 5.3
ṀHE = = = 4105.26ton.
The water that is trapped by the molecular sieve is calculated from equation 5.2
W= ṀHE - ṀAE=4105.26-3,945 =160.26 ton of water.
Material balance on distillation
Distillation is used for recovery of bioethanol from water-bioethanol mixture. The mixture of
bioethanol water which obtained from fermentation process is separated using two-stage
distillation column to recovered 96% bioethanol from the mixture of 3.50% bioethanol.

D=4105.26ton
D=? 2nd XD1=0.96
Distillation
F=? XD= 0.60
ZF= 0.035 1st

Distillation
W1=?
Xw1 =0.99
W=?
Xw=1
Figure 5.3 Material balance on distillation
Where:
 F = feed to the 1st distillation
 D=distillate from 1st distillation
 ZF =feed composition of light component
 D1= distillate from 2nd distillation
 W= bottom product
Over all mass balance on the 2nd distillation:
D = D1+W1
D = 4105.26 +W1….……………………………………………………………………… (5.4)
Bioethanol mass balance on the 2nd distillation:
0.60D=0.96D1 +0.01W1…………………………..……………………………………… (5.5)
Substitute equation 5.4 in to 5.5
0.6(4105.26 +W1) = 0.96*4105.26 +0.01W1
W1=2505 ton
D=4105.26 + 2505 =6610 ton

Over all mass balance on the 1st distillation:

F = D+W
F= 6610 +W…………………………………………………………………………………. (5.6)
Bioethanol mass balance on the first distillation:

0.035 F = 0.60D +0…………………………..………………………………………………. (5.7)
Substitute equation 5.6 in to 5.7
F=187669 ton and W=173145 ton.
Material Balance on Fermentation
Material balance in the fermenter is carried out based on the fermentation reaction.
C6H12O6 2C2H5OH +2CO2
180 g/ mol 92g/mol bioethanol + 88 g/mol CO2 for 100% conversion.
Mass of filtrate (MF) =? Mass of CO2 release (Mco2) =?
Mass of fermented mash
Mass of media (Mm) =? (MFm) = 187669 ton
Figure 5.4 Material balance on fermenter

Assume 90% glucose can converted into bioethanol if the fermentation process very effective.
0.90MG= (4105.26 x0.96x180)/92

MG= 8567.5 ton of glucose is required annually.
Where
HE = hydrous ethanol
MG = mass of glucose
The amount of carbon dioxide produced is;
MCO2 = (8,567.5 x88)/180

= 4188.55 ton of CO2 released per year.

Over all mass balance on the fermentation process:
MF +MM = MFM + MCO2
MM = 0.1MF
MF +0.1MF = 187669 ton + 4188.55 ton
MF = 174415.98 ton of filtered Hydrolysate is input to the fermenter.
Mm = 17441.60 ton of inoculum is required.
Material balance on pressurized bag filter
This is used for separating insoluble solid (lignin) from liquid (sugar solution). Moisture content
of sludge (lignin) assumed to be 24.5%.
Hydrolysate (Mh) mass of filtrate (Mf)= 174416 ton

Xs = 0.044 XL=1
XL=0.956
Lignin (Ml), Xs=0.755, XL=0.245
Figure 5.5 Material balance on filter

Mh=Ms +Mf………………………………………………………………………………….. (5.8)
Mass balance on the solid
Mh *0.044 =Ms *0.755
Mh=17.16 Ms………………….…………………………………………………………… (5.9)
Substitute equation (5.9) into equation (5.8) to calculate mass of lignin or total solid remain after
hydrolysis
16.16 Ms =Mf
Ms = (174416ton)/16.16
= 10793 ton of solid or lignin was generated per year.
The amount of hydrolysate is calculated from equation (5.9)

MH=185209 ton of hydrolysate is need per year.

Material balance in dilute acid treatment (Hydrolysis)
The mass balance was based on the general plant operation of bioethanol production from
literature obtained; the liquid solid ratio was fixed at 8:1, in the first stage hydrolysis 1.25%w/w
acid concentration is used to decomposed hemicellulose and in 2 nd stage 1.68 %w/w acid
concentration is utilized to decomposed cellulose in to its monomer which is called glucose. The
amount of solid charged in to the 2nd stage is 60% of the total solid charged into the 1st stage and
remain 40% is decompose in the 1st stage hydrolysis.
Hydrolysis reaction is
C6H10O5 + H2O H2SO4 + C6H12O6
162g/ mole of cellulose 180g/ mole of glucose for 100% conversion
But not 100% conversion and assume that 95% of cellulose converts to glucose. According
Dehnavi, (2009) 38.8% % of corn cob is cellulose.
MA1 (1.25%)
Mcc 1st
Stage
MC =0.39(Mcc) MH =185209 ton
0.60Mcc MG= 8568 ton
2nd
MA2 (1.67%)
Stage
Figure 5.6 Material balance on hydrolysis reactor

By using stoichiometry equation the feed dry corn cob (Mcc) is calculated as;
0.39*0.95* Mcc = (162* 8568)/180

Mcc = 20813ton dry corn cob is feed per year.
Mass balance on the 1st stage:
The liquid solid ratio in the 1st stage is (8:1)

MA1=8*Mcc
MA1= 166504ton per year of 1.25% diluted sulfuric acid is required per year in the 1st stage
hydrolysis. The acid required in this stage is 208 ton/year.
The same is true for the 2nd stage hydrolysis; liquid solid ratio was (8:1)
MA2=8*0.60Mcc
MA2= 99902 ton per year of 1.67% diluted sulfuric acid is required per year in the 2nd stage
hydrolysis. The acid required in this stage is 1668 ton/year.
Mass balance on dryer
MWcc Mcc =20813ton

Dryer
Xw=0.484 Xw=0.10
Xs=0.516 Xs=0.90
Mw
Figure 5.7 Material balance on dryer

Mwcc =Mw + Mcc
Component mass balance on the solid cc
0.516 Mwcc = 0.90 Mcc
Mwcc= (0.90*20813)/0.516 = 36302 ton of wet corn cob is feed to the dryer.
From the above over all mass balance, mass of water is 15489 ton.
5.3 Energy balance
The energy balance is performed based on mass balance results.
The basic of the calculation is one hour.
T=25oC T=70oC
Dryer
M= 36302ton

Figure 5.8 Energy balance on dryer

The required energy is calculated as
Q = m Cp ΔT
Where
Q= required energy
M=mass of wet corn cob
Cp = CPw*Xw + Cps *Xs
= 4.18*0.484 +4.18*0.44*0.516 = 2.97kJ/kg .k
Q = m Cp ΔT
= 36302*2.97 (70-25) = 4815676.8KJ/year = 594.58KJ/hr.
Energy balance on hydrolysis
The energy used for hydrolysis process is generated from water steam.
MA1= 20405kg/hr.
T=25oC T1=121oC
Mcc1=4445 kg/hr. 1st

Stage
MH=22697k/hr.
Mcc2=1530kg/hr.
MA2=12243kg/hr. 2nd
Stage
T2=140oC
Figure 5.9 Energy balance on hydrolysis reactor

Where
Mcc1=mass of cc input to 1st hydrolysis stage
Mcc2=mass of cc input to 2nd hydrolysis stage
MA1=mass of water and acid solution input to 1st stage hydrolysis
MA2= mass of water and acid solution input to 2nd stage hydrolysis
The amount of heat required for the hydrolysis process
Q = M1Cp1ΔT1 + M2Cp2ΔT2
Cp1= = 4.11kJ/kg.K.

Cp2 = =4.14kJ/kg.K
Q=24850 *4.11(121-25) + 13773 *4.14 (140-25) = 16362141kJ /hr.

Energy balance on Fermentation
Mm=2137kg/hr.
MF=21374 kg/hr. MFm=22999kg/hr.
Fermenter
Mco2=513 kg/hr.
Figure 5.10 Energy balance on fermenter

Exothermic energy is released during fermentation; the amount of heat generated can be
calculated. The outlet temperature is 30oC
Cp of mix at 30oC = 4.14kJ/kg k
Cp of CO2 at 30oC = 0.846kJ/kg k
QMIX = QCO2 + QFh+ Q
Q = QMIX - QCO2 - QFM
Q= Mmix*Cpmix ΔT-MCO2CpCO2 ΔT-MFm*Cp*Fm *ΔT
CpFm =Cpmix*Xmix+Cpco2*Xco2
Xco2=513/ (21374+2137) =0.0218
XFm=1-0.0.0218=0.978
CpFm =3.781kJ/kg .k
Q =23511.4*4.14*(30-25)-513*0.846*(30-25) - 22999*3.781(30-25)
= 49,826.61kJ/hr.

Energy Balance on Distillation

Basis; 25oC and one hour
D=503 kg
Xde=0.96
F=22,999kg Xdw=0.04, T=25oC
XFW=0.979
1st
XFE=0.035 Distillation
T=30oC
W=22,496 kg, Xw=1, Xe=0
Figure 5.11 Energy balance on distillation column

Specific heat capacity of distillate (Cpd)
Cp = 0.96*2.72+0.04* 4.18=2.778kJ/kg k
Specific heat capacity of the bottom product is specific heat capacity of water
Cpw =4.18kJ/kg k
Mass balance on the condenser, assume reflux ratio i.e. R=L/D=2.5
L=2.5D=2.5*503kg/hr = 1257.5 kg/hr
V=L+D=1257.5 kg/hr + 503 kg/hr = 1760.5 kg/hr
From equilibrium data, boiling point of ethanol at 96% is 78.13oC
At steady state condition
Input = Output
HF = HD + HL +QC
QC=HF-HD-HL
Assume complete condensation, therefore
Enthalpy of Vapor = Latent + Sensible heat
HV = mvλv + mvcpΔT
=1760.5 * 789+1760.5 * 2.778(78.13-25) =1,648,876 kJ/hr
Overall balance to determine QB
Input = Out put

QB + HF = QC + HD + HW
QB=QC+HD+HW-HF
HF = mFCpΔT
Cp=0.021*2.72+0.979*4.18 = 4.15 kJ/kg k
HF=22999 kg/hr*4.15*(30-25) = 477229.25 kJ/hr
HW = mwCpΔT=22,496 *4.18 (100-25) =7,052,496 kJ/hr
QB = QC + HD + HW - HF
QB=1,648,876 kJ/hr + 0 + 7,052,496 kJ/hr - 477229.25 kJ/hr =8,224,143 kJ/hr
Amount /mass of steam required
Latent heat of steam at 274KN/m2, λv=2174kJ/kg, therefore mass of steam required,
QB=Ms* λs
Ms=QB/ λs = (8,224,143 kJ/hr) / (2174 kJ/kg) = 3783 kg/hr.
Mass of water that is able to condense is removed with temperature rise of 30oC
QC=Mw Cp ΔT, M = = = 13149 kg/hr.

Energy balance on molecular sieve
Molecular sieve is a mass transfer unit operation that is used to adsorb a small constituent
molecule of water which is present in the ethanol to increase the concentration of bioethanol and
raise the concentration of bioethanol to around 99.9%.
Mass of water =19.64 kg
Xw=1, Xe=0%
Hydrous bioethanol
(ṀHE) = 503kg
XHE=96%
XHE=96% Anhydrous bioethanol (ṀAE) =483.3kg
Xw=4% XAE=99.9%, T=95oC
To=25oC Xw=0.01%
Figure 5.12 Energy balance on molecular sieve

Cp is calculated based on their percentage of the input mass of hydrous bioethanol
Cp = 0.96*2.72+0.04*4.18 = 2.78kJ/kg k

The temperature entering the molecular sieve is 25oC and exit is at a temperature of 95oC
Q =M Cp ΔT
= 503* 2.78*(95-25)
= 97884kJ.

6 SIZING OF MAJOR EQUIPMENTS
Assumption:
 All tanks are 85% full or 15% safety factor.
 Sizing of equipment is depending on the material balance calculated on the above section.
 Basis: one day.
Sizing of dried corn cob storage tank
 Material of construction: carbon steel
 Material handled: dried corn cob
 Density of corn cob: 282kg/m3
 Required mass per day: 88,415 kg
Density = mass/volume
The required volume of storage tank is for 3 days.
Volume dried corn cob = (88,415 kg/day)/282kg/m3
= (314m3)*3=941m3. This the volume occupied by the corn cob.
Vt = (941/0.85) = 1107m3. This is the capacity of the tank that holds the corn cob for 3 days.
Sizing of water storage tank
 Material of construction: carbon steel
 Material handled: water
 Density: 1000 kg/m3
 Mass of water: 1,115,453kg/day
Volume of water = = 1115 m3.
Volume of vessel = (1115.45m3)/ (0.85)

= 1312.29 m3.
Sizing of fermenter
 Material construction: carbon steel
 Material to be handled: Hydrolysate
 Shape: dome shaped
 Density of liquid: 880kg/m3.
 Temperature: 300C
 Mass of feed: 564288 kg/day.

The residence time of one batch is 72 hour or 3 days, so that three fermenters are required for the
ensuring fermentation days.
V= = 641m3.
Required volume of vessel with 15% safety facto r is
Vvessel = = 754m3.
Sizing of bioethanol Storage tank

 Material of constriction: Carbon steel
 Material handled: bioethanol
 Density 0.789kg/lit: 789kg/m3
 Temperature of liquid: 200C
 Mass of bioethanol per day is 11603kg/day.
V= = 15m3. This is the volume occupied by bioethanol.
Required volume of vessel;
Vvessel = = 17m3.
The product may be stay for around one week or 7 days, so that the total volume of vessel
required for one week is 122 m3.
Sizing of distillation column
Specification
 Mode of operation: continuous
 Degree of separation 96% bioethanol and 4% water @ the distillate
 Contacting device: plate type
 Operating pressure: 0.5 bar
 Vapor velocity: 0.75 m/s
 Feed flow rate: 1780 kg/h or 81 kmol/h
 Feed composition 60 % bioethanol and 40% of water
 Boiling point of bioethanol = 78.13oC = 351.13k
 Boiling point of water = 100oC = 373k
 Reflux ratio = 2.5.
Over all mole balance:
F=D+B

81 kmol/h =D + B
Bioethanol mole balance:
81*0.60 = 0.96D + 0.01B
Combine the two equations and we got
D = 50.3 kmol/h and B = 30.7 kmol/h
L/D= 2.5
V= L + D =3.5 D =176.05 kmol/h.
Molecular weight of feed = 0.6*46 + 0.4 * 18 = 34.8 g/mol
Mole fraction of bioethanol in the feed of distillation (XEF)
XEF= = 0.37.
( )
Mole fraction of water in the feed of distillation (XWF)

XWF =1-XEF = 0.63.
Mole fraction of bioethanol in the distillate (XED)
XED = = 0.904.
( )
Mole fraction of water in the distillate (XWD)

XWD= 1-XED =0.096.
Water mole fraction in the bottom (XwB)
XwB = = 0.996.
( )
Bioethanol mole fraction in the bottom (XEB)

XEB = 1-XWB = 0.004.
Specific heat of feed (Cpf) = 0.6*2.72 +0.4*4.18= 3.304 kJ/kg. K
Molar latent heat of the feed is
λ= 0.60*790 + 0.40*2376.7
= 1424.68kJ/kg
=1424.68 kJ/kg*(1kg/0.021kmol) = 67842.9kJ/kmol
= 67841.9 kJ/kmol*0.23 kcal/kJ
=1503.64 kcal / kmol
Average boiling point of feed

= ΣXiTi
= 0.37*308 + (1- 0.37)*308 = 308k.
Heat capacity of the feed
Cpf =3.304 kJ/kg. K *0.23kcal/kmol
=0.760 kcal/kg. K.
Vapor liquid equilibrium data of bioethanol water solution given below in table below
Table 6.1 Vapor liquid equilibrium data of bioethanol water solution
X(Liq) 0.0190 0.0721 0.0966 0.1238 0.1661 0.2337 0.2608 0.3273 0.3965 0.5079
Y(vap) 0.1700 0.3891 0.4375 0.4704 0.5089 0.5445 0.5580 0.5826 0.6122 0.6564
X(Liq) 0.5198 0.5732 0.6763 0.7472 0.8943

Y(vap) 0.6599 0.6841 0.7385 0.7815 0.8943
Quality of feed (q)
q= = = 1.
The slope of the q-line = = = ∞.
The slope of the operating line for the rectifying section = = 0.714.
Y-intercept of operating line for the rectifying section is =XED/(R+1) =0.904/ (2.5+1) =0.26.
From the graph below, the operating line for the stripping section is drawn to pass through the
point x = xB = 0.004 on the 45o line and join to the point of intersection of the q-line and the
operating line for the rectifying section.
With the above information, number of theoretical plate is 7 and feed is introduced at the 5
stages.
Number of stages:
Number of stages =theoretical plate – 1=7-1= 6 thus 4 stages are on the rectifying section and 2
stages are present on the stripping section.
Diameter of distillation column:
Assume diameter of column is constant in the stripping and rectifying section. Temperature at
the top is 85oC (358k) and Assuming an ideal gas behavior solution

Figure 6.1 McCabe-Thiele diagrams to determine number of stage
V= , where n is molar flow rate of vapor n =81.76 kmol/hr.
R= 0.082atm.m3/kmol. K.
P= 0.5atm.
V= = 2400 m3/hr.
Cross sectional area;
A= = = 0.889m2.
Diameter of column
D=√ (4As/Π) =1.06 m.
Pump for delivering of water
 Type: centrifuge pump
 Head: 4m

 Slurry to be handled: distilled water

 Temperature of liquid: 25oC
 Materials of construction: carbon steel
 Capacity: 0.04m3/s.
Pump for delivering hydrolysate
 Head: 4m
 Slurry to be handled: hydrolyzate
 Capacity: 0.004m3/s.
Pump for delivering filtered hydrolysate
 Head; 4m
 Slurry to be handled: hydrolyzate
 Density: 888kg/m3
 Capacity 0.004m3/s.

Flow sheet of bioethanol production from corn cob

Figure 6.2 Flow sheet of bioethanol production from corn cob

7 PRELIMINARY ENGINEERING ECONOMIC ANALYSIS
7.1 Total Capital Investment
Total Capital Investment = Fixed Capital Investment + Working Capital Investment

For this case, capital investment items are calculated based on the purchased equipment cost of
the plant.
Fixed capital cost = ƒ (purchased equipment cost).
Table 7.1 purchased cost of equipment
Equipment name capacity quantity unit price ($)

Drum drier 30m2 2 134980
Dry cob storage tank 1107m3 1 76661
3
Fermenter 754 m /batch 3 67708
Water St. Tank 1312m3 2 95552
Filter(plate and frame) 16 m2 2 31374
Mixing tank 26 m3 2 36454
Hydrolysis reactor 15 m3 2 88906
Hydrolyzate St. Tank 2185m3 1 134288
Neutralizer 755 m3 1 67708
bioethanol St. Tank 122 m3 1 33142
Distillation No. stage=7 2 58894
Molecular sieve 45.27kg/hr 1 6000
3
ST yeast propagation 75.5m 1 48042
H2SO4 St. Tank 8.00m3 1 47008
NaOH St. Tank 5.00m3 1 42308
Centrifugal pump 0.004m3/s 4 46810
Diaphragm pump 0.025m3/s 2 15725
Total 1,458,484
Data Sources: http://www.mhhe.com/engcs/chemical/peters/data/

Estimation of fixed capital investment

Table 7.2 estimation of fixed capital investment
Direct cost (DC) Percentage (%) Price ($)

Purchased equipment E 1,458,484
Purchased equipment installation 25%E 364,621
Instrumentation and control 10%E 145,848
Piping (installed) 15%E 218,773
Electrical( installed) 10%E 145,848
Building (including service) 11%E 160,433
yard improvement 5%E 72,924
Service facilities 35%E 510,469
A. Total direct cost (TDC) 3,077,400
Indirect cost (IC)
Engineering and Supervision(14%E) 204,188
Construction expenses and Contractor’s fee(15%E) 218,773
Contingency(8%E) 116,679
B. Total Indirect cost (IC) 539,640
Fixed-capital investment (A+B) 3,617,040
TCI=FCI+WC, since working capital cost is (10-20) % of total capital investment

TCI =FCI+0.15TCI
TCI = FCI/0.85= $4,255,341
Working cost (WC) = $(4,255,341-3,617,040) = $638301
7.2 Total production cost
Total production cost =manufacturing + general expense

Manufacturing cost =direct production cost +fixed charge +plant overhead cost
General expense =administrative +distribution and selling costs+ interest

Table 7.3 Total cost of raw materials
Raw materials Capacity (tons) Unit price ($) Total cost ($)
Corn cob 52,433 2.38 124,790
H2SO4 5,416 200 1,083,200
NaOH 295 350 103,250
Total raw material cost 1,309420
A. Manufacturing cost
Manufacturing cost = Direct production costs (DPC) + Fixed charges (FC) + Plant overhead
costs (POC)
a. Direct production cost (60% TPC)
1. Raw material and inputs = $1,309,420

2. Operating labor (10% of TPC) = 0.10 TPC
3. Direct supervisor and clerical labor (12% of operating labor) = 0.10 * 0.12 * TPC = 0.012
TPC
4. Utilities (15% of TPC) = 0.15 TPC
5. Maintenance and repair (6% of FCI) = $3,617,040* 0.06 = $217,022
6. Operating supplies = 0.5% FCI = 0.005* $3617040 = $18085
Total direct production cost =1,546,347 + 0.272 TPC
b. Fixed Charges
1. Depreciation (10% FCI) = $361704

2. Insurance (1% FCI) = $36170
3. Local taxes (2%FCI) = $72341
Total fixed charges = $470215
c. Plant Overhead costs
1. 5% of total product cost =0.05 TPC
Total Manufacturing cost = a + b + c
= 2016562+0.322 TPC
B. General expenses
 Administrative cost (2% of total product cost) = 0.02 TPC
 Distribution and selling cost (2% of total product cost) = 0.02 TPC

Total general expenses = 0.04 TPC

Total product cost = manufacturing cost + general expenses
TPC=$2016562+0.322 TPC +0.04 TPC
TPC (1-0.362) =$2016562
TPC= $3,160,755
7.3 Economic evaluation
Gross earn cost
Current price of 99.9% ethanol $0.85/lit based on the current price of world biofuel market
Annual revenue=$0.85/lit X 5,000,000 lit
=$4,250,000
Gross annual profit = Annual revenue –Total production cost
=$(4,250,000-3,160,755)
=$1,089,245.
Net income
Net income= Gross annual profit –income tax (35 % Gross annual profit)
=$(1,089,245-1,089,245*0.35)
=$718,902.
Percent of profit
% profit= *100%
=23%.
Percent rate of return
The yearly profit divided by the total initial Investment necessary represents Return on
Investment. Taking the risk factor Mar = 12%, to be the plant feasible RoI >Mar (must).
Net income =$718,902
Total capital investment (TCI) = $4,255,341
Rate of return, % ROR = *100% = 17% > 12% it is acceptable.
Payback period
The minimum length of time theoretically necessary to recover the original fixed capital
investment in the form of cash flow is called payback period.

Assume 10 years Project service life and we use straight line method to calculate depreciation.
Dep. = = $361,704.
Payback period = =3.3 year < 5 year, it is acceptable.
Break-even point (BEP)

At BEP
Total production cost = total sales income
Direct production cost +fixed charges + plant overhead cost + General expense = Total sales
income
n*unit direct production cost + fixed charges + plant overhead cost + General expense = n*unit
sale income
Where
n= production capacity per year
Unit direct production cost =Total production cost / Total production capacity
Unit direct production cost = $3,160,755/5,000,000 liter/year..
=$0.63/liter.
Unit sale income = $0.85/liter
n*$0.63 +$470215 +126,430.2 + 158,038 = $0.85*n
n= 3,430,377 liter
Break-even production capacity = 68.6% of the total production capacity.

8 CONCLUSION AND RECOMMENDATIONS
8.1Conclusion
Production of bioethanol from biomass was one way to reduce both consumption of crude oil
and environmental pollution. The main difficulty in bioethanol production from conventional
sources is that raw material availability was limited; on the other hand 75% food-material
inflation (worldwide) is attributed to using conventional feedstock for bioethanol production. As
biofuels are very essential for the environment and the economy when they were produced from
lignocellulosic biomass, selection of the cheap and appropriate raw material was big task.
This study examines the possibility of corncob for bioethanol production. The conversion of
corncob to bioethanol was carried out with dilute acid pretreatment, dilute acid hydrolysis,
fermentation and distillation process steps.
Corn cob is promising lignocellulosic feedstocks for production bioethanol fuel. It was the most
abundant by-product generated from the maize crop. Bioethanol production from such
lignocellulosic material was carried out in four main stages such as pretreatment, hydrolysis (first
and second stage), fermentation and distillations.
In this study two-stage diluted acid hydrolysis was used and the effect of the hydrolysis process
variable (temperature, time and acid concentration) in the yield of bioethanol was investigated
and optimized using response surface methodology. Positive yield of ethanol was obtained at a
high acid concentration and low temperature as well as at high temperature and low acid
concentration.
Based on the rough economic analysis, production of bioethanol from corn cob was profitable
since the rate of return on investment was 37%, this shows us the project returns its 37% of the
initial investment in one year and the payback period is around four years.

8.2 Recommendation
Based on the current study the following recommendations are forwarded:

 Further researches have to be carried out to increase the yield of bioethanol from corn
cob by using other microorganisms which are capable of converting 5- and 6- carbon
sugar into bioethanol.
 Detailed economic feasibility studies in the production process are recommended, since it
is critical for the rationale of commercialization.
 Further optimization of the Pretreatment, Fermentation and Distillation processes are
recommended to maximize the yield of bioethanol from corn cob.
 Corn cob is rich in cellulose and hemicelluloses, due to this it is used as a low valuable
raw material in different applications, such as for production of xylitol, furfural, biogas
and bioethanol. But in developing country this valuable by-product is used as low-cost
animal feed and the remaining are deposited into landfills. Further research is required to
utilize this largely available by-product, corn cob.
 Producing bioethanol from renewable resources is becoming an important issue for the
whole world. Therefore, the work needs to be continued for scaling up of bioethanol
production from corn cob.
 Dilute acid was used for hydrolysis of cellulose; however hydrolysis with enzyme has to
be checked.

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comparison of liquid hot water and steam pretreatments of sugar cane bagasse
bioconversion to ethanol, Bio-resource Technology 81, 33–44.
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Third Eddition , Washington DC. pp 637.

APPENDIX
Appendix A: Density and Yield of Ethanol

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MEKELLE UNIVERSITY

DEPARTMENT OF CHEMICAL ENGINEERING

STREAM: BIOCHEMICAL ENGINEERING

COURSE TITLE: RESEARCH METHODS AND PROPOSAL WRITING

COURSE CODE: CHEG251

PROJECT TITLE: PRODUCTION OF BIOETHANOL FROM CORN COB

Submission Date: 12/06/2011E.C

Department of Chemical Engineering Page i

Department of Chemical Engineering Page ii

3.3 Dilute Acid Hydrolysis .................................................................................................... 23

Department of Chemical Engineering Page iii

Department of Chemical Engineering Page iv

Table 2.1 Major Maize Producing Administrative Zones of Ethiopia ...........................................6

Department of Chemical Engineering Page v

LIST OF ABRREVATIONS AND ACRONYEMS

Cp Specific heat capacity

Department of Chemical Engineering Page vi

Keywords: Bioethanol, corn cob, diluted-acid hydrolysis, and Saccharomyces cerevisiae

Department of Chemical Engineering Page vii

Lignocellulosic biomass is a cheap, renewable, abundantly available resource (Hansen et

Department of chemical engineering Page 1

Department of Chemical Engineering Page 2

1.3.1 General objective

The specific objectives of the project are the following;

Department of Chemical Engineering Page 3

Department of Chemical Engineering Page 4

Department of Chemical Engineering Page 5

Table 2.1 Major Maize Producing Administrative Zones of Ethiopia

Zone Average production (in quintals)

Table 2.2 Physico-chemical characteristics of bioethanol

No. parameter Characteristic properties

Department of Chemical Engineering Page 6

2.3 Bioethanol feedstock’s

Department of Chemical Engineering Page 7

Exploitation of first generation feedstocks for future bio-fuel production is ultimately

Department of Chemical Engineering Page 8

Department of Chemical Engineering Page 9

Figure 2.2 Schematic illustration of the cellulose chain

Hemicellulose or polyose is a mixture of polymers comprising pentoses, hexoses hexuronic acids

Lignin is a complex, hydrophobic, cross-linked, three-dimensional aromatic polymer of phenyl

Department of Chemical Engineering Page 10

2.3.3 Third generation feedstocks

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2.5 Bioethanol Production Process

Biological conversion consists of exposing biomass to certain microorganisms. The secondary

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 High yield of hemicelluloses sugar,

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Department of Chemical Engineering Page 14

C6H12O6 2C2H5OH + 2CO2 ……………………………………………………….... (2.3)

One of the most successful microorganisms for bioethanol production is Saccharomyces

Department of Chemical Engineering Page 16

Department of Chemical Engineering Page 17

2.5.2 Thermochemical Process

Department of Chemical Engineering Page 18

 It provides a much greater energy yield.

Department of Chemical Engineering Page 19

Department of Chemical Engineering Page 20

The materials used to run all experiments were listed below:

Department of Chemical Engineering Page 21

Moisture content (%) = *100

Where: W1= Initial weight, W2= weight after drying

Volatile content (%) =

Ash content (%) = =

0.390.95 Mcc = (162* 8568)/180

Q=24850 4.11(121-25) + 13773 4.14 (140-25) = 16362141kJ /hr.