Bio Ethanolll Final PDF

DEBRE BERHAN UNIVERSITY DEPARTMENT OF CHEMICAL ENGINEERING
Acknowledgement
Beyond all we thank our almighty GOD who allows us to do this project and helped us to
accomplishment of this study.
Next, we would like to acknowledge our advisors instructor Gedefaw A and Kidist G. for high
guidance, advise, patience and encouragement in this work. We also would like to express our
gratitude to chemical engineering staff and laboratory technicians for their support and positive
advice during this work.
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page i

Table of contents
Contents page
Acknowledgement ........................................................................................................................... i
List of figures ................................................................................................................................. vi
List of tables .................................................................................................................................. vii
List of Acronyms ......................................................................................................................... viii
Abstract ........................................................................................................................................... x
1. INTRODUCTION ...................................................................................................................... 1
1.1 Background of the study ....................................................................................................... 1
1.2. Statement of the problem ..................................................................................................... 2
1.3. Objectives of the study ......................................................................................................... 2
1.3.1. General objective ........................................................................................................... 2
1.3.2. Specific objective .......................................................................................................... 2
1.4. Significance of the study ...................................................................................................... 2
1.5. Scope of the project .............................................................................................................. 2
1.6. Limitation ............................................................................................................................. 3
2. LITERATURE REVIEW ........................................................................................................... 4
2.1. History of ethanol production .............................................................................................. 4
2.2. Bio-ethanol and its application ............................................................................................. 4
2.3. Ethanol and its characteristics .............................................................................................. 7
2.4. Feedstocks for bioethanol production .................................................................................. 8
2.4.1. Bioethanol from lignocelluloses feedstocks .................................................................. 9
2.5. Barley spent grain............................................................................................................... 11
2.5.1. Physicochemical properties of barley spent grain (BSG) ............................................ 11
2.5.2. Potential applications for barley spent grain ............................................................... 12
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page ii

2.6. Bioethanol production process ........................................................................................... 13
2.6.1. Pretreatment ................................................................................................................. 13
2.6.2. Hydrolysis .................................................................................................................... 17
2.6.3. Fermentation ................................................................................................................ 19
2.6.4. Distillation ................................................................................................................... 20
2.6.5. Dehydration ................................................................................................................. 20
2.7. Technology selection.......................................................................................................... 22
2.8. Factors affecting hydrolysis of lignocellulosic materials ................................................... 23
2.8.1. Properties of the substrate............................................................................................ 23
2.8.2. The acidity of the system ............................................................................................. 23
2.8.3 Rate of decomposition of hydrolysis products during hydrolysis ................................ 24
2.9. Factors affecting fermentation ........................................................................................... 24
2.9.1. Effect of sugar concentration ....................................................................................... 24
2.9.2. Effect of temperature ................................................................................................... 24
2.9.3. Effect of pH ................................................................................................................. 25
2.9.4. Ethanol concentration .................................................................................................. 25
2.9.5. Osmotic tolerance ........................................................................................................ 25
2.9.6. Inhibitor tolerance........................................................................................................ 26
2.10. Previous study of bioethanol production .......................................................................... 26
3. MATERIAL AND METHODS ................................................................................................ 29
3.1. Materials and equipment used for the experiments ............................................................ 29
3.2. Experimental set up ............................................................................................................ 30
3.3. Experimental procedure ..................................................................................................... 31
3.3.1. Sample preparation ...................................................................................................... 31
3.3.2. Dilute acid pretreatment .............................................................................................. 31
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page iii

3.3.3. Filtration ...................................................................................................................... 32
3.3.5. Sterilization.................................................................................................................. 33
3.3.6. Fermentation ................................................................................................................ 33
3.3.7. Ethanol Separation (Distillation) ................................................................................. 34
4. RESULT AND DISCUSSION ................................................................................................. 36
4.1. Moisture content determination ......................................................................................... 36
4.2. Ethanol characterization ..................................................................................................... 36
4.2.1. Specific gravity of ethanol ........................................................................................... 36
4.3. Effect of yeast concentration .............................................................................................. 37
4.4. Effect of fermentation time ................................................................................................ 39
5. MATERIAL AND ENERGY BALANCE ............................................................................... 41
5.1 Material balance .................................................................................................................. 41
5.2. Energy balance ................................................................................................................... 47
6. SIZING AND DESIGN OF MAJOR EQUIPMENTS ............................................................. 52
6.1. Sizing.................................................................................................................................. 52
6.2. Design of process equipment ............................................................................................. 56
7. FINANCIAL EVALUATION .................................................................................................. 67
7.1. Cost estimation ................................................................................................................... 67
7.2. Feasibility Study (economics) ............................................................................................ 71
8. PLANT LOCATION AND PLANT LAY OUT ..................................................................... 76
8.1. Site selection ...................................................................................................................... 76
8.2. Plant lay out ........................................................................................................................ 76
9. ENVIRONMENTAL CONSIDERATION .............................................................................. 78
9.1. General Considerations ...................................................................................................... 78
10. CONCLUSION AND RECOMMENDATION...................................................................... 80
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page iv

10.1. Conclusion........................................................................................................................ 80
10.2. Recommendations ............................................................................................................ 81
References ..................................................................................................................................... 82
Appendices .................................................................................................................................... 85
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page v

List of figures
Figure 2.1: Structure of cellulose .................................................................................................. 10
Figure 2.2: structure of lignin ...................................................................................................... 10
Figure 2.3: BSG supply chain material flow ............................................................................... 11
Figure 2.4: Schematic representations on Biomass Pre treatment (Mosier et al, 2004) ............... 17
Figure 2.5: Production flow diagram of bioethanol ..................................................................... 21
Figure 2.6: Technological selection ............................................................................................. 23
Figure 2.7: Annual ethanol productions by the major producers, adapted from Mabee (2007) .. 27
Figure 3.1: Laboratory experimental set up .................................................................................. 30
Figure 3.2: Bioethanol production flow sheet from barley spent grain ....................................... 35
Figure 4.1: Yeast concentration vs specific gravity ...................................................................... 38
Figure 4.2: yeast concentration vs alcohol content ...................................................................... 39
Figure 4.3: Specific gravity vs fermentation time ....................................................................... 40
Figure 4.4: Fermentation time vs Alcohol content ...................................................................... 40
Figure 5.1: Laboratory scale material balance .............................................................................. 41
Figure 8.1: plant lay out ............................................................................................................... 77
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page vi

List of tables
Table 2.1: Physico - chemical characteristics of ethanol as liquid fuel ......................................... 7
Table 2.2: Composition of barley spent grain (Dehnavi) ............................................................ 12
Table 2.3: World fuel ethanol production by country or region (million Gallons) ..................... 27
Table 2.4: Ethanol production in Ethiopia .................................................................................... 28
Table 4.1: Moisture content determination ................................................................................... 36
Table 4.2: Volume, density and alcohol concentration of ethanol .............................................. 37
Table 4.3: Effect of yeast concentration and the fermentation at 3 day ...................................... 38
Table 4.4: Effect of Yeast concentration on alcohol content ........................................................ 39
Table 4.5: effect of fermentation time on specific gravity............................................................ 39
Table 4.6: effect of fermentation time on alcohol content ........................................................... 40
Table 5.1: Annual current and future demand of fuel ethanol in (litter)...................................... 41
Table 6.1: Specification of dried spent grain storage tank............................................................ 52
Table 6.2 Specification of water storage tank ............................................................................... 52
Table 6.3: Specification of hydrolysis reactor .............................................................................. 53
Table 6.4: Specification of hydrolyzates storage tank ................................................................. 54
Table 6.5: Specification of fermenter .......................................................................................... 54
Table 6.6: Specification of ethanol storage tank .......................................................................... 54
Table 7.1: Equipment cost estimation........................................................................................... 67
Table 7.2: Estimation of fixed capital investment ........................................................................ 69
Table 7.3: Raw material ............................................................................................................... 70
Table 7.4: Operating labor ........................................................................................................... 70
Table 7.5: selling price determination ......................................................................................... 71
Table 7.6: Determination of payback period ............................................................................... 72
Table 7.7: cash flow ...................................................................................................................... 74
Table 7.8: determination of present value.................................................................................... 75
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page vii

List of Acronyms
AFER Ammonia Fiber Explosion
BEP Breakeven point
BEV Breakeven volume
BSG Barley spent grain
Cp specific heat capacity
D mass flow rate of hydrous ethanol
ID Inner diameter
OD Outside diameter
DC Direct cost
E Purchased Equipment cost
f design stress
F Mass flow rate dilute hydrous ethanol
FCI Fixed capital investment
GE General Expense
GP Gross profit
HD Enthalpy of Distillate
HF Enthalpy of Feed
HL Enthalpy of liquid refluxed
HV Enthalpy of Vapor flowing up
IC Indirect cost
IRR Internal rate of return
J welding efficiency
M(S-w) Mass flow rate of Soft water
MAE Mass flow rate of anhydrous ethanol
MHE Mass flow rate of hydrous ethanol
MM Mass flow rate of Media
MPTM Mass flow rate of pretreated mixture
MS Mass flow rate of Steam
MSA Mass flow rate of dilute sulfuric acid
MW Mass flow rate of water
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page viii

MC Manufacturing cost
MPa Mega Pascal
NP Net profit
NPV Net present value
PBP Payback period
pH potential of hydrogen
PV Present value
QC Heat of condenser
ROR Rate of return
rpm revolution per minute
t wall thickness of tanks
TCI Total capital investment
TDC Total direct cost
TPC Total production cost
VLE Vapor liquid equilibrium
WV Weight of vessel
WC Working Cost
WCI Working capital investment
D diameter of cylindrical section
α half of the angle of the cone
S the hypothnuios of the conical section
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page ix

Abstract
The objective of this study was production of bioethanol from Barley spent grain by using two-
stage diluted acid hydrolysis. The production process was carried out in four main stage such as
pretreatment, hydrolysis (first and second stage), fermentation and distillations. In first stage
diluted acid hydrolysis the process variables were fixed at the best optimum condition
liquid/solid ratio of 8 g/g, 0.1 ml H2SO4 /g of dry matter, 17 min reaction time and 121o C
temperature. Before the first stage hydrolysis process was carried out, the wet Barley spent grain
was dried using oven at 110oC temperature for 20hr. 336ml of distilled water and 5.88 ml of
sulfuric acid were added to non- soluble component that obtained from first stage acid hydrolysis
and after drying at 120oc for 18hr.The sample was then hydrolyzed in the autoclave with 140oC
for 25min. After hydrolysis separate the solid particle from liquid. Fermentation of the
hydrolyzate was performed using 50ml,100ml,150ml propagated yeast (Saccharomyces
cerevisiae) at 30oCtemperature, pH 4.5 and 3day and 6 day fermentation time for all samples.
After fermentation and distillation the specific gravity of the produced alcohol was measured
by Pycnometer (specific gravity bottle) and alcohol concentration was obtained from the
relationship between the specific gravity and the proportion of alcohol concentration in
ethanol.
As a result the best operational condition obtained at 100ml of yeast concentration and 3 day
fermentation time, which is 48.3% alcohol concentration using simple distillation. Once the
experimental work was completed, scaling up is made to design a plant that produces
ethanol with the capacity of 400,000 litter per year. The economical evaluation shows that the
plant needs 4,648,036.53 ETB total capital investment cost. The feasibility study of the project
proves that the project is viable with ROR of 27.5 % ,BEP of 68.34%, a NPV 4,732,658.25 ETB
and the project life is 10 years, it recover itself in 3.7 years.
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page x

1. INTRODUCTION
1.1 Background of the study
Bioethanol as fuel has been known over hundred years. In 1860 Nicholas August Otto from
Germany employed ethanol as fuel in his internal combustion engine. From beginning of last
century up to 1960 mixed ethanol with gasoline was used widely for transportation in many
European countries such as Germany, France, Italy, Sweden, England as well as Brazil and
U.S.A. In 1960s the interest to use of ethanol decreased due to low price of oil (in
comparison with ethanol price). The new interest in Bioethanol once more started in
many countries (Brazil in 1979, U.S.A. in 1980 and Europe in 1990) owing to
technological developments, market factors and some other factors such as national energy
security concern and governmental motivations. Nowadays a huge amount of ethanol is
produced around the world. For instance, in 2008 the U.S.A. was the first fuel ethanol producer
with 9,000 millions of gallons. In that year Brazil with 6,472.2 and the European Union with
733.6 millions of gallons fuel ethanol were the second and third producer of fuel ethanol
in the world respectively. The main raw materials in those countries are sugar or starch (corn,
wheat, sugar cane and sugar beets). Nowadays, concerns about if there will be enough corn to
support the demand for both fuel and food is increasing. Therefore the competition between
food and ethanol industries may lead the price of grain and sugar in the future (Sun and Cheng
2002).
Energy consumption has increased steadily over the last century as the world’s population
increases, and more and more countries become industrialized. The traditional source fossil fuel
is continuously being depleted irrespective of the new geographical discoveries (Campbell and
Lahrrere 1998) estimated the current known oil reserve as well as the reserves yet to be
discovered and concluded that the world crude oil production will begin to decline in 2010.
There have also been concerns about the pollution and various health risks associated with the
use of petroleum as fuel. In view of these, the importance of alternative energy source has
become even more necessary not only due to the continuous depletion of the limited fossil fuel
stock but also for safe and better environment (Chandel, et al. 2007).
BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page 1

1.2. Statement of the problem

Due to the potential exhausting of traditional fossil fuels and the increasing price of petroleum
together with environmental concerns, the search for alternative renewable fuels has
attracted great attention in recent years.
Now days, the numbers of brewery industries are increasing in Ethiopia and they generate a
high amount of barley spent grain. This by-product is mainly used as cattle feed but it contains
starch that used for bioethanol production. So the Conversion of such by product into
bioethanol helps to reduce energy problem of developing countries like Ethiopia.
1.3. Objectives of the study

1.3.1. General objective
The general objective of the study was production of bioethanol from barley spent grain.
1.3.2. Specific objective
To investigate effect of fermentation time and yeast concentration on bio ethanol production.
To characterize bioethanol obtained from distillation process.
To perform Size of main equipments.
To evaluate the economic feasibility.
1.4. Significance of the study
 Producing bio ethanol from barley spent grain (BSG) has a significant advantage for
energy source and decreases the impact of greenhouse gas.
 Bioethanol production from BSG is considered as 2nd generation biofuel process since it
has no direct conflict with human food.
 The cost of bioethanol will be decreased. This can increase the consumption of ethanol in
the country.
 Creates energy security to our country.
 It creates work opportunity for peoples in our country.
1.5. Scope of the project
This project work ranges from literature survey, developing of methodology, performing
experiment to produce bio ethanol, plant design that contains market study, material balance and
energy balance, specification, sizing and apparatus design of main equipments and to perform
economic analysis.

1.6. Limitation
During the laboratory analysis there were so many problems like:
 Absence of shaking incubator during fermentation. Due to the absence of shaking
incubator it was difficult to manage the temperature.
 Absence of alcoholmeter to measure the alcohol content of product.
 Absence of simple distillation hinders the ethanol from reaching to the desired
concentration.
 Shortage of chemicals for yeast propagation (DAP, Dexterous sugar) limits experimental
runs.

2. LITERATURE REVIEW
2.1. History of ethanol production
Ethanol is an alcohol made through the fermentation of plant sugars from agricultural crops and
biomass resources (NEVC, 1998). With rapid depletion of the world reserves of petroleum,
ethanol 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.
Ethanol was one of the most popular lamp illuminants used in 1850s and approximately 90
million gallons ethanol was produced in the United States. But due to the tax imposition on
ethanol to assist in financing the civil war and the cheaper price of kerosene, it quickly
replaced ethanol as the premier illuminant 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 the 20th 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). It’s market grew from less than a billion litres in 1975 to more
than 39 billion litres in 2006 and reach approximately 100 billion litres in 2015 (Licht, 2006).
2.2. Bio-ethanol and its application

Bioethanol is ethanol that is produced from biological materials (biomass).It is water-free
alcohol produced from the fermentation of sugar or converted starch. It is colourless clear
Liquid with mild characteristic odour. It is volatile, miscible in both water and non-polar solvents
at ordinary conditions (Purwadi 2006).

It is applicable as:-
 Fuel
Engine fuel
Ethanol, also known as ethyl alcohol with the chemical formula C2H5OH is a flammable, clear,
colorless and slightly toxic chemical compound with acceptable odour. It can be produced either
from petrochemical feedstocks by the acid-catalyzed hydration of ethene, or from biomass
feedstocks through fermentation. On a global scale, synthetic ethanol accounts for about 5-10%
of total production while the rest is produced from fermentation of biomass mainly sugar crops,
e.g. cane and beet, and of grains (Licht 2006).
Ethanol as a neat fuel or even in the blended form with gasoline has a long history as automotive
fuel. In 1860, German inventor Nicholas Otto used ethanol as a fuel in an early prototype of an
internal combustion engine because it was widely available throughout Europe for use in spirit
lamps. A few years later, Henry Ford built his first automobile with an engine that could run on
ethanol. In 1908, Ford unveiled his Model T engine equipped with carburetors that could be
adjusted to use alcohol, gasoline or a mixture of both fuels (Solomon 2007).
However, fossil fuels were predominantly used for automobile transportation throughout the last
century, obviously due to their lower production cost. As an automotive fuel, hydrous ethanol
can be used as a substitute for gasoline in dedicated engines. Anhydrous ethanol, on the other
hand, is an effective octane booster when mixed in blends of 5% to 30% with no engine
modification requirement (Licht 2006).
Rocket fuel
Ethanol was commonly used as fuel in early bipropellant rocket (liquid propelled) vehicles, in
conjunction with an oxidizer such as liquid oxygen. The German V-2 rocket of World War II ,
credited with beginning the space age, used ethanol, mixed with 25% of water to reduce the
combustion chamber temperature (Braeuning 2006).The V-2's design team helped develop U.S.
rockets following World War II, including the ethanol-fueled Redstone rocket which launched
the first U.S. satellite (Abrief history of rocketry n.d.). Alcohols fell into general disuse as more
efficient rocket fuels were developed (Braeuning 2006).

Fuel cells
Commercial fuel cells operate on reformed natural gas, hydrogen or methanol. Ethanol is an
attractive alternative due to its wide availability, low cost, high purity and low toxicity. There are
wide range of fuel cell concepts that have been trialled including direct-ethanol fuel cells, auto-
thermal reforming systems and thermally integrated systems. The majority of work is being
conducted at a research level although there are a number of organizations at the beginning of
commercialization of ethanol fuel cells (Badwal, et al. 2015).
 Medical purpose
Antiseptic
Ethanol is used in medical wipes and most common antibacterial hand sanitizer gels as an
antiseptic. Ethanol kills organisms by denaturing their proteins and dissolving their lipids and is
effective against most bacteria and fungi, and many viruses. However, ethanol is ineffective
against bacterial spores (McDonnel 1999).
Medicinal solvent
Ethanol, often in high concentrations, is used to dissolve many water-insoluble medications and
related compounds. Liquid preparations of cough and cold remedies, pain medication, and mouth
washes may be dissolved in 1 to 25% concentrations of ethanol and may need to be avoided in
individuals with adverse reactions to ethanol such as alcohol-induced respiratory reactions
(Admas and Rans 2013). Ethanol is present in over 700 liquid medications including
acetaminophen, iron supplements, ranitidine, furosemide, mannitol, Phenobarbital, trimethoprim/
sulfamethoxazole and over-the-counter cough medicine (Zuccotti and Fabiano 2011).
Recreational purpose
As a central nervous system depressant, ethanol is one of the most commonly consumed
psychoactive drugs (Alcohol use and safe drinking n.d.).The amount of ethanol in the body is
typically quantified by blood alcohol content (BAC), which is here taken as weight of ethanol
per unit volume of blood. Small doses of ethanol, in general, produce euphoria and relaxation;
people experiencing these symptoms tend to become talkative and less inhibited, and may exhibit
poor judgment. At higher dosages (BAC > 1 g/L), ethanol acts as a central nervous system
depressant , producing at progressively higher dosages, impaired sensory and motor function,
slowed cognition, stupefaction, unconsciousness, and possible death. Ethanol is commonly
consumed as a recreational drug, especially while socializing, due to its psychoactive effects.
 Household heating
Ethanol fireplaces can be used for home heating or for decoration. (Can ethanol Fire places Be
cozy 2016).
 Feedstock
Ethanol is an important industrial ingredient. It has wide spread use as a precursor for other
organic compounds such as ethyl halides , ethyl esters, diethyl ether, acetic acid, and ethyl
amines .
 Solvent
Ethanol is miscible with water and is a good general purpose solvent. It is found in paints,
tinctures, markers, and personal care products such as mouthwashes, perfumes and deodorants.
However, polysaccharides precipitate from aqueous solution in the presence of alcohol, and
ethanol precipitation is used for this reason in the purification of DNA and RNA.
2.3. Ethanol and its characteristics

Bioethanol or fuel alcohol refers to ethyl alcohol produced by microbial fermentation (as
opposed to petrochemically-derived alcohol) that is used as a transportation biofuel. It is
produced through distillation of the ethanolic 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 summaries some of the important characteristics
of ethanol as a fuel source.
Table 2.1: Physico - chemical characteristics of ethanol as liquid fuel
Parameter Characteristic properties
Molecular formula C2H5OH
Molecular mass 46.07 g/mol
Appearance Colourless liquid
Water solubility (between –117°C and 78°C)∞ (miscible)
Density 0.789 kg/l

Boiling temperature 78.5°C (173°F)
Freezing point –117°C
Flash point 12.8°C(lowest temperature of ignition)
Ignition temperature 425°C
Explosion limits 19%(v/v) Lower 3.5% (v/v) Upper
Vapour pressure @ 38°C 50 mm Hg
Higher heating value (at 20°C) 29,800 KJ/kg
Lower heating value (at 20°C) 21,090 KJ/kg
Specific heat Kcal/Kg 60°C
Acidity (pKa) 15.9
Viscosity 1.200 mPa.s (20°C)
Refractive index (nD) 1.36 (25°C)
Octane number 99
Source: (Walker, 2011)
2.4. Feedstocks for bioethanol production

Ethanol can be produced synthetically from petroleum or by the microbial fermentation
of sugars. The three main groups of raw materials for the production of ethanol by fermentation
are sugar, starch, and lignocellulose (Lin and Tanaka 2006). Sugar-containing raw materials
include sugarcane, sugar beets, fruits and sweet sorghum. Starch is a biopolymer and defined as a
homopolymer consisting only one monomer, D-glucose (Pongsawatmanit, Temsiripong and
Suwonsichon 2007). During bioethanol production from starch, it is necessary to break
down the chains of this carbohydrate for obtaining glucose syrup, which can be converted
into bioethanol by yeasts. This type of feedstock is the most utilized for bioethanol production
in North America and Europe (Sanchez, et al. 2004). Starch can be converted to fermentable

sugar by a method called the hydrolysis technique. Hydrolysis is a reaction of starch with
water, typically performed by cooking the starch at high and low temperatures which is normally
used to break down the starch into fermentable sugar. Dextrin oligosaccharides are generated by
adding α-amylase and glucoamylase to obtain glucose (Wheals, et al. 1999).
Lignocellulose-based feedstocks that can be considered for ethanol production are wood
residues, agricultural residues, and the spent sulfite liquor from pulp and paper mills.
The advantage of using lignocellulose as raw material for ethanol production is due to its
abundance and relatively cheap (Wheals, et al. 1999).In contrast to sugar-containing crops,
the utilization of lignocellulose as a substrate for ethanol production is difficult or
relatively recalcitrant to hydrolysis (Gray, Zhao and Emptage 2006). Because of its complex
structure, this resists degradation. The basic structure of all lignocellulosic biomass consists of
cellulose (C6H10O5) x, hemicelluloses (C5H8O4) m, and lignin [C9H10O3.(OCH3)0.9 -1.7]n.
Production of ethanol from lignocellulosic waste materials, such as sawdust from the forestry
industry and BSG from Beer industries have a benefits from the fact that the energy input for
the overall process can be kept low. Energy-related expenses for planting, fertilization and
harvesting can be avoided if waste materials are used. If lignocellulosic waste materials are used,
there will not be any competition for the limited agricultural land available, which might be
needed for food production (Sun and Cheng 2002).
2.4.1. Bioethanol from lignocelluloses feedstocks

Lignocelluloses feedstocks consists of three main components namely cellulose, Hemicelluloses
and lignin.
Cellulose
Cellulose is a polysaccharide of hundreds or thousands of molecules of glucose with the
formula (C6H10O5) n. Cellulose molecules consist of long chains of glucose molecules like
starch molecules with a different structural configuration the encapsulation of cellulose the
more difficult to hydrolyze than starch polymers (Harinen 2004).

Figure 2.1: Structure of cellulose
Hemicellulose
Hemicelluloses include long chains of sugar molecules consisting of: (1) five-carbon sugars (D-
xylose and L-arabinose) and (2) six-carbon sugars (D-galactose, D-glucose and D-mannose) and
(3) uronic acids. In comparison with cellulose, hemicelluloses are amorphous and relatively easy
to hydrolyze. During hydrolysis, the hemicellulose from hardwoods releases products high in
xylose (a five-carbon sugar), but the hemicellulose contained in softwoods yields more six-
carbon sugars (Harinen 2004).
Lignin
Lignin is a complex polymer of phenyl propanoid units that are not fermented. Lignin also is
resistant to chemical and enzymatic degradation and typically hardwood contains less lignin than
softwood (Harinen 2004)
Figure 2.2: structure of lignin

2.5. Barley spent grain

Barley spent grains are lignocellulosic materials, which are the main by-product of the brewing
industry corresponding to around 85%of the total by product.
This by-product contains high amounts of protein and fiber, but till now has been limited mainly
for animal feed, human nutrition and in some case as substrate for microorganisms with different
Figure 2.3: BSG supply chain material flow

proposes such as cultivation of microorganism and enzyme production. In addition, some
investigation on the enzymatic hydrolysis of spent barley grain has been done in order to
release of ferulic and p-coumaric acids with an esterase and a xylanase. Although a huge
amount of this byproduct produces every year, a few investigations have done to consider it as a
low-cost raw material for energy production. The spent barley grain usually include of the husk
of the original barley grain, but depending on original raw materials used for bear production
and also harvesting time, additive and process condition could contain different composition
(Mussatto and Roberto, 2006).
2.5.1. Physicochemical properties of barley spent grain (BSG)

Barley spent grain basically consists of the husk–pericarp–seed coat layers that covered the
original barley grain. Depending on the evenness of malting more or less starchy endosperm and
walls of empty aleurone cells may also remain. The starch content will be negligible, and some
residues of hops introduced during mashing will be present depending on the brewing regime
used. Thus, the major components of BSG will be the walls of the husk–pericarp–seed coat,

which are rich in cellulose and non-cellulosic polysaccharides and lignin, and may contain some
protein and lipid. The husk also contains considerable amounts of silica and much of the
polyphenolic components of the barley grain (Macleod, 1979). According to (Kunze 1996), 25%
of the minerals present in barley are present as silicates. The bright points in the external portion
of the husk are silicates. The chemical composition of BSG varies according to barley variety,
harvest time, malting and mashing conditions, and the quality and type of adjuncts added in the
brewing process (Huige, 1994; Santos et al., 2003).
But in general, BSG is considered as a lignocellulosic material rich in protein and fiber, which
account for around 20 and 70% of its composition, respectively. Microscopic examination shows
the presence of numerous fibrous tissues from the surface layers the original barley grain. The
main components of these fibrous tissues are arabinoxylans, lignin (a polyphenolic
macromolecule) and cellulose (a linear homopolymer of glucose units). Analyses of BSG are
shown in Table 1. Santos et al. (2003) found, besides fibre, 24.2% protein, 3.9% lipid and 3.4%
ash in oven-dried BSG. Table 2shows the protein, apparent starch and non-starch polysaccharide
composition, and monosaccharide analyses of the non-starch polysaccharide fraction of BSG
from pilot scale trials with malting barley and a feed barley. Protein and fiber are highly
concentrated in spent grain because most of the barley starch is removed during mashing (Kissel
and Prentice, 1979).
Table 2.2: Composition of barley spent grain (Dehnavi)
Component % of dry material
Hemicellulose 32.5
Cellulose 27.6
Lignin 13.4
Water 10.2
2.5.2. Potential applications for barley spent grain

Although Barley spent grain is the main by-product of the brewing process, it has received little
attention as a marketable commodity, and its disposal is often an environmental problem.
Nevertheless, due to its chemical composition it can be of value as a raw material. Some possible
applications for this agro-industrial by-product are described below.
Animal nutrition
Human nutrition
Energy production
Charcoal production
Paper manufacture
Adsorbent
Substrate for enzyme production
Hydroxycinnamic acids (ferulic and p -coumaric) extraction
2.6. Bioethanol production process

The major unit processes involves in the production of bioethanol from Lignocellulosic material
are pretreatment, hydrolysis, fermentation, and distillation.
2.6.1. Pretreatment
Pre-treatment is a vital step in production of ethanol from lignocellulosic materials. It is
required in order to change the structure of cellulosic materials and to make cellulose more
accessible to the enzymes, which convert the carbohydrate polymers into fermentable sugars.
During pre-treatment of lignocellulosic materials, many component can be formed that act as
inhibitor for enzymatic hydrolysis and cellular growth. Therefore selection of the best method
for a certain raw material is very important. There are lots of pretreatment methods for
lignocellulosic materials, but each method has advantages and disadvantages and so that
depending on feedstock, the best one should be selected. Besides, the optimum reaction
parameters of the various pretreatments, such as temperature, pressure, and reaction time, are
specific for each type of feedstock and so that they should be optimized for specified raw
material (Parveen 2009).
The main parameters that contribute to the resistance of lignocellulosic materials to hydrolysis
include: accessible surface area, crystallinity of biomass, lignin protection, and covering by
hemicellulose. Therefore the main aim of the pretreatment step is removing lignin and
hemicellulose, decreasing the crystallinity of cellulose and enhancing the porosity. In other

words, the best method for pretreatment must meet the following key necessities: (1) more sugar
formation, (2) less degradation of carbohydrates, (3) less formation of inhibitors, (4) less cost
and energy consumption. It should be noted that high yield of sugars does not always result in
high conversion to ethanol, because lignocellulosic components or chemicals used in
pretreatment may form compounds that inhibit fermentation. Pretreatment methods are classified
in to; physical such as milling/grinding, physicochemical like steam pretreatment/auto-
hydrolysis, hydro-thermolysis/wet oxidation, chemical such as alkali/dilute acid/oxidizing
agents/organic solvents, biological, electrical and combination of them (Jin Seop Bak 2009).
2.6.1.1. Chemical Pretreatment Method

Acid catalyzed: Concentrated acids such as H2SO4 and HCl have been used to treat
lignocellulosic materials. Although they are powerful agents for cellulose hydrolysis,
concentrated acids are toxic, corrosive and hazardous and require reactors that are resistant to
corrosion. In addition, the concentrated acid must be recovered after hydrolysis to make the
process economically feasible. Dilute acid has been successfully developed for pretreatment of
lignocellulosic materials. The dilute sulfuric acid pretreatment can achieve high reaction rates
and significantly improve cellulose hydrolysis (Nirbhay Gupta 2008).
At moderate temperature, direct saccharification suffered from low yields because of sugar
decomposition. High temperature in dilute acid treatment is favorable for cellulose hydrolysis.
There are primarily two types of dilute acid pretreatment processes: high temperature (greater
than 160°C), continuous-flow process for low solids loading (5–10% (weight of substrate/weight
of reaction mixture)), and low temperature (less than 160 °C), batch process for high solids
loading (10–40%). Although dilute acid pretreatment can significantly improve the cellulose
hydrolysis, its cost is usually higher than some Physico-chemical pretreatment processes. A
neutralization of pH is necessary for the downstream acidic or enzymatic hydrolysis or
fermentation processes (Nirbhay Gupta 2008).
Alkaline Pretreatment: Some bases can also be used for pretreatment of lignocellulosic
materials and the effect of alkaline pretreatment depends on the lignin content of the materials.
The mechanism of alkaline hydrolysis is believed to be saponification of intermolecular ester
bonds cross linking xylan hemicelluloses and other components, for example, lignin and other

hemicellulose. The porosity of the lignocellulosic materials increases with the removal of the
cross links. Dilute NaOH treatment of lignocellulosic materials caused swelling, leading to an
increase in internal surface area, a decrease in the degree of polymerization, a decrease in
crystallinity, separation of structural linkages between lignin and carbohydrates, and disruption
of the lignin structure. The digestibility of NaOH-treated hardwood increased from 14% to 55%
with the decrease of lignin content from 24–55% to 20%. However, no effect of dilute NaOH
pretreatment was observed for softwoods with lignin content greater than 26%. Dilute NaOH
pretreatment was also effective for the hydrolysis of straws with relatively low lignin content of
10–18% used the combination of irradiation and 2% NaOH for pretreatment of corn stalk,
cassava bark and peanut husk. Ammonia was also used for the pretreatment to remove lignin and
an ammonia recycled percolation process for the pretreatment of corn cobs/Stover mixture and
switch grass. The efficiency of delignification was 60–80% for corn cobs and 65–85% for
Switch grass (Nirbhay Gupta 2008).
Oxidative Delignification: Lignin biodegradation could be catalyzed by the per oxidases
enzyme with the presence of H2O2. The pretreatment of cellulosic biomass with hydrogen
peroxide greatly enhanced its susceptibility to enzymatic hydrolysis. About 50% lignin and most
hemicellulose were solubilized by 2% H2O2 at 30 °C within 8 h, and 95% efficiency of glucose
production from cellulose was achieved in the subsequent saccharification by cellulases at 45 °C
for 24 h (Nirbhay Gupta 2008).
2.6.1.2. Biological pretreatment

In biological pretreatment processes, microorganisms such as brown, white and soft-rot fungi are
used to degrade lignin and some hemicellulose in waste materials. Brown rots mainly attack
cellulose, while white and soft rots attack both cellulose and lignin. White-rot fungi are the most
effective basidiomycetes for biological pretreatment of lignocellulosic materials. The
pretreatment of wheat straw by 19 white-rot fungi and found that 35% of the straw was
converted to reducing sugars by Pleurotus ostreatus in five weeks. Similar conversion was
obtained in the pretreatment by Phanerochaete sordida and Pycnoporus cinnabarinus in four
weeks (Nirbhay Gupta 2008).
The white-rot fungus P. chrysosporium produces lignin-degrading enzymes, lignin peroxidases
and manganese-dependent peroxides, during secondary metabolism in response to carbon or

nitrogen limitation. Both enzymes have been found in the extracellular filtrates of many white rot
fungi for the degradation of wood cell walls. Other enzymes including polyphenol oxidases,
laccases, H2O2 producing enzymes and quinine reducing enzymes can also degrade lignin. The
advantages of biological pretreatment include low energy requirement and mild environmental
conditions. However, the rate of hydrolysis in most biological pretreatment processes is very low
(Nirbhay Gupta 2008).
2.6.1.3. Physico-chemical pretreatment

Ammonia Fiber Explosion (AFEX): AFEX is Physico-chemical pretreatment in which
lignocellulosic materials are exposed to liquid ammonia at high temperature and pressure for a
period of time and then pressure is swiftly reduced. The concept of AFEX is similar to steam
explosion. In a typical AFEX process, the dosage of liquid ammonia is 1-2 kg ammonia /kg dry
biomass, temperature 90ºC, and residence time 30 minutes. AFEX pretreatment can significantly
improve the saccharification rates of various herbaceous crops and grasses. To reduce the cost
and protect to environment, ammonia must be recycled after the pretreatment. The ammonia
pretreatment does not produce inhibitors for the downstream biological processes, so water wash
is not necessary. AFEX pretreatment does not require small particle size for better efficiency
(Holtzapple and A 1990).
Steam Explosion (Auto hydrolysis): Steam explosion is the most commonly used method for
the pretreatment of lignocellulosic materials. In this method, chipped biomass is treated with
high-pressure saturated steam and then pressure is swiftly reduced, which makes the materials
undergo an explosive decompression. Steam explosion is typically initiated at a temperature of
90 - 160 ºC for several minutes to a few hours the material is exposed to atmospheric pressure.
The process causes hemicellulose degradation and lignin transformation due to high temperature,
thus increasing the potential of cellulose hydrolysis and steam treatment with dilute acid of 0.5 to
1%, temperature of 90 to 140oC and residence time of 10 to 40 minutes gives highest yield of
ethanol (Solomon 2007).
The advantages of steam explosion pretreatment include the low energy requirement and no
recycling or environmental costs. Limitations of steam explosion include destruction of a portion
xylan fraction, incomplete disruption of the lignin-carbohydrate matrix and generation of

compounds that may be inhibitory to microorganisms used in downstream processes. Because of

the formation of degradation products that are inhibitory to microbial growth (McMillan 1994).
Figure 2.4: Schematic representations on Biomass Pre treatment (Mosier et al, 2004)
2.6.2. Hydrolysis
In the hydrolysis reaction, the complex chains of sugars that make up the hemicellulose are
broken, releasing simple sugars. The complex hemi-cellulose sugars are converted to a mix of
soluble five-carbon sugars, xylose and arabinose, and soluble six-carbon sugars, mannose and
galactose. The rest of hemicelluloses are degraded to weak acids, furan derivates, and phenolic.
These compounds, however, are potential fermentation inhibitors (Cellulosic Ethanol, 2010).By
the action of acids and or enzymes, the glucose yields of cellulose hydrolysis often exceed 90%,
but hydrolysis without preceding pretreatment yields typically less than 20% only. The
pretreated feedstock can be hydrolyzed by two methods (Cellulosic Ethanol 2010).
Acid hydrolysis: Acids have been used to catalyze (speed up) the hydrolysis of starch in “starch
cookers operating at temperatures of 50 to 150°C, a process referred to as acid hydrolysis. Acid
pretreatment for ethanol production was developed in Germany in the 19th century. In the United
States, Forest Service, Forest Products Laboratory (FPL) conducted extensive research using
acid pretreatment for ethanol production from wood (Zhu et al., 2008).There are two basic types
of acid processes: dilute acid and concentrated acid, each with variations. Dilute acid processes
are conducted under high temperature and pressure, and have reaction times in the range of
seconds or minutes while concentrated acid process conducted under low temperatures and
pressures employed allow the use of relatively low cost materials such as fiberglass tanks and
piping (Zhu et al., 2008).

Dilute Acid Processes: Most dilute acid processes are limited to a sugar recovery efficiency of
around 60 to 75%. The reason for this is that at least two reactions are part of this process. 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. Thus, once the cellulosic molecules are broken
apart, the reaction proceeds rapidly to break down the sugars into other products most notably
furfural, a chemical used in the plastics industry. Not only does sugar degradation reduce sugar
yield, but the furfural and other degradation products can be poisonous to the fermentation
microorganisms (Mohammad, 2008).
The biggest advantage of dilute acid processes is their fast rate of reaction, which facilitates
continuous processing. For rapid continuous processes, in order to allow adequate acid
penetration, feedstocks must also be reduced in size so that the maximum particle dimension is in
the range of a few millimeters. 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 process conditions to recover the 5-carbon sugars while the second stage is
conducted under harsher conditions to recover the 6-carbon sugars. Unfortunately, sugar
degradation is still a problem and yields are limited (Cellulosic Ethanol 2010).
Concentrated Acid Process: Concentrated acid process uses relatively mild temperatures and
the only pressures involved are usually only those created by pumping materials from vessel to
vessel. One concentrated acid process was first developed by USDA and further refined by
Purdue University and the Tennessee Valley Authority. In the TVA concentrated acid process,
corn Stover is mixed with dilute (10%) sulfuric acid, and heated to 100ºC for 2 to 6 hours in the
first (or hemicellulose) hydrolysis reactor. The low temperatures and pressures minimize the
degradation of sugars. To recover the sugars, the hydrolyzed material in the first reactor is
soaked in water and drained several times. The solid residue from the first stage is then
dewatered and soaked in a 30% to 40% concentration of sulfuric acid for 1 to 4 hr as a
precellulose hydrolysis step (Cellulosic Ethanol 2010).
This material is then dewatered and dried with the effect that the acid concentration in the
material is increased to about 70%. After reacting in another vessel for 1 to 4 hr at 100ºC, the
reactor contents are filtered to remove solids and recover the sugar and acid. The sugar/acid
solution from the second stage is recycled to the first stage to provide the acid for the first stage
hydrolysis. The sugars from the second stage hydrolysis are thus recovered in the liquid from the
first stage hydrolysis (Cellulosic Ethanol 2010).The primary advantage of the concentrated
process is the high sugar recovery efficiency, which can be on the order of over 90% of both
hemicellulose and cellulose sugars. The low temperatures and pressures employed allow the use
of relatively low cost materials such as fiberglass tanks and piping. Unfortunately, it is a
relatively slow process and cost effective acid recovery systems have been difficult to develop.
Without acid recovery, large quantities of lime must be used to neutralize the acid in the sugar
solution. This neutralization forms large quantities of calcium sulfate, which requires disposal
and creates additional expense (Cellulosic Ethanol 2010).
Enzyme hydrolysis: is a method in which cellulases are utilized for the hydrolysis. This is a
quite new approach compared to concentrated-acid and dilute-acid hydrolysis. Cellulolytic
enzymes were discovered during World War II when American scientists found the agent that
was responsible for army clothing deterioration in the jungles of the South Pacific. The organism
responsible for producing the cellulolytic enzymes was Trichoderma reesei, which today is used
in the enzyme industry for producing a wide range of commercial enzymes (Sheehan and
Himmel).The cellulases involved in the hydrolysis of lignocellulose include endoglucanases,
which attack low-crystallinity regions of the cellulose fiber and generate free chain-ends, and
exoglucanases, which remove cellobiose from the free chain ends. Then, Jglucosidase hydrolyses
cellobiose to glucose (Sun and Cheng 2002).
The advantages of enzymatic hydrolysis are high yields, due to the highly specific cellulose
conversion, and that the reaction is performed at moderate temperatures. Furthermore, the
byproduct formation is low. The disadvantages are the slow reaction rate of the enzymes and the
high enzyme cost (Lynd et al., 2005).
2.6.3. Fermentation
Fermenting microorganisms are used for the conversion of monomeric sugars to ethanol.
Different organisms such as bacteria, yeast and fungi can be used for the conversion, however
the most frequently used organism in industrial processes are the robust yeast
Saccharomyces cerevisiae (baker‟s yeast). Under anaerobic condition S.cerevisiae produces
ethanol from hexose as the overall shows below. The drawback of Saccharomyces cerevisiae is
not produce ethanol from xylose but Pichia stipitis yeast is produce ethanol from both

glucose and xylose sugar, however this type of yeast also has many drawbacks such as
low ethanol ,temperature and inhibitor resistance (Galbe and Zacchi 2002).
C6H12O6 yeast 2C2H5OH + 2CO2 + 3C
5H10O5 yeast 5C2H5OH+ 5CO2
2.6.4. 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. This means that proportion between two materials, in this case ethanol and
water can be changed, and still, there are two materials in layers, the liquid and the vapor layers
(Onuki 2005).
2.6.5. Dehydration
After distillation, about 5% of water remains in ethanol. Especially, this water is a big problem
for fuel ethanol because the presence of this amount of water enhances the molecular polarity of
ethanol 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 ethanol 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 and another method is azeotropic distillation.
Molecular sieves
There is a lower bound on the fraction of ethanol entering the molecular sieve (0.8). Adsorption
takes place at 95°C. Heat exchanger heats the inlet stream from the mixer up to 95°C. The
molecular sieve is a bed of zeolite that operates in semi-continuous mode. The bed is saturated
with water after a period of time and is then regenerated. Hence, there are usually two sieves
being operated in parallel – one being saturated with water while the other is being regenerated
(or dehydrated) using air under vacuum. Heat exchanger heats air with an assumed relative
humidity of 70% at 20 °C to 95 °C. The air at the outlet of the dehydrating molecular Sieve is
cooled down to 25 °C in heat exchanger and this stream leaves this exchanger saturated with
water at 25 °C (Mariano Martin and Ignacio E. Grossmann, Carnegie Mellon University).
BSG
Drying
Steam pretreatment
Filtration
Drying
Dilute acid Hydrolysis
Filtration
Neutralization
Fermentation
Distillation Molecular sieve
Water Bottom product
Figure 2.5: Production flow diagram of bioethanol

2.7. Technology selection

Bio Ethanol production technology can be classified in to:-
1) Biochemical conversion
2) Thermo chemical conversion
1) Biochemical conversion :-in which enzymes and other micro-organisms are used to
convert cellulose and hemicellulose components of the feedstocks to sugars prior to their
fermentation to produce ethanol
2) Thermo chemical conversion:-where gasification technologies produce a synthesis gas (CO+
H2) from which a wide range of long carbon chain biofuels, such as synthetic diesel or
aviation fuel can be reformed.
The technology selected in this study is biochemical conversion because it proceed at Lower
temperature and less coasty than thermo chemical conversion.
Sample Preparation
The sample that acquire must be prepared and conditioned for pretreatment, hydrolyze,
fermentation and distillation. Sample preparation process include: drying (by oven dryer),
milling (by using ball mill) and sieving after the samples were collected.
Pre –Treatment
The purpose of the pretreatment was to remove lignin, 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, and avoid the formation of
By product inhibitors and must be cost effective. Based on this dilute sulfuric acid pretreatment
is used in this process.
Acid Hydrolysis
The cellulose molecules which are composed of long chains are broken down to simple sugar,
before it is fermented for alcohol production. Even though there are many types of hydrolysis
types, dilute acid hydrolysis is an easy, fast rate of reaction and also the amount of alcohol
produced in case of acid hydrolysis is more than that of alkaline hydrolysis so chose dilute
sulfuric acid hydrolysis.

Lignocelluloses Milling Pretreatment Biochemical conversion
Hydrolysis Fermentation Ethanol
Gasification Fermentation Ethanol

Thermo chemical conversion
Catalytic conversion
Figure 2.6: Technological selection
2.8. Factors affecting hydrolysis of lignocellulosic materials

2.8.1. Properties of the substrate
The properties of the substrate can affect the hydrolysis. These properties are: neutralizing
capacity, proportion of easily hydrolysable hemicellulose and cellulose, amount and rate of
hydrolysis of the difficult to hydrolyze materials, the length of the macromolecules, degree of
polymerization of cellulose, configuration of the cellulose chain, and association of cellulose
with other protective polymeric structures within the plant cell wall such as lignin, pectin,
hemicellulose, protein, mineral elements,etc. Particle size is also one of the effective parameters.
2.8.2. The acidity of the system

Another parameter affecting the hydrolysis is the acidity of system. The acidity is dependent on
the type and concentration of the acid used, amount of acid solution, amount of acid
released from the biomass during hydrolysis, liquid to solid ratio, the neutralizing capacity of
the lignocellulose and movement of the solution during heating. When dilute acid hydrolysis is
applied in a continuous process such as a screw fed co-current reactor, it requires relatively short
residence time. Therefore, the penetration of acid catalyst in the biomass, as well as dispersion
in the reactor, can significantly affect the overall reaction, and consequently the reactor
performance. Diffusivity of sulfuric acid is dependent on the nature of the lignocellulosic
material (Demirbas 2005).

2.8.3 Rate of decomposition of hydrolysis products during hydrolysis

The rate of decomposition of the products during the hydrolysis process depends on
temperature, acidity, reaction time and the concentration of sugars. Under hydrolysis
conditions that produce a solution containing in excess of 10 percent glucose, reversion
phenomena are suggested to be very important. The reversion phenomena result in much of the
glucose being present not as free glucose but as dimmers, oligomers, and anhydrosugars
which are unavailable to the microorganism used in fermentation (Harris, Baker and Zerbe
1984). It was recently reported that metals and or metal ions can also catalyze glucose
decomposition during acid hydrolysis of Lignocellulosic materials.
2.9. Factors affecting fermentation
Microorganisms for ethanol 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, osmotic tolerance, specificity, yield, genetic stability,
and inhibitor tolerance (Demirbas 2005).
2.9.1. Effect of sugar concentration
The concentration of sugar can affect the microbial ethanol fermentation in various ways. Use of
concentrated sugar substrate is one of the ways to obtain high ethanol yield during
fermentation. The amount of ethanol 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, Pamment and Greenfield 1981).
2.9.2. Effect of temperature
Temperature has an important factor on the growth rate of the microorganisms and the rate of
ethanol production. Wine and beer fermentations are generally conducted below 20°C, 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).
All the recombinant strains are mesophilic organisms and have best function between 30 to
38°C. Operating at greater temperatures is desirable for the following reasons:
High fermentation temperature increases growth rate and productivity exponentially
Plant capital cost is less due to higher productivity per unit volume of ferment or
vessel and cooling equipment investment is lowered.
Operating costs are less since less energy is required to maintain desired fermentation
temperature and recover the ethanol.
Contamination risk is less as fewer organisms exist at high temperatures.
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).
2.9.3. 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 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).
2.9.4. Ethanol concentration
Concentration of ethanol in the fermentation broth can directly affect the growth rate of the
culture and its ability to convert sugar to ethanol. Inhibitory and toxicity level of ethanol vary
from culture to culture. Higher temperature lowers the tolerance of the organism. At
temperatures above 35 °C, current strains lose viability at ethanol concentrations of 10 %
(w/v) (Hettenhaus 1998).
2.9.5. Osmotic tolerance
The semi-permeable membrane surrounding the cell must be able to withstand wide osmotic
pressure changes in extracellular fluids that impact the relative osmotic pressure difference. If
not, the cells may be severely damaged or even killed. The cells may burst in a hypotonic
solution, when the solution becomes more dilute than the intracellular fluid. If hypertonic, the

cells will shrink from the osmotic pressure difference. Osmotic pressure limits can be one of the
factors that restrict maximum substrate concentration (Sofer and Zaborsky 1981).
2.9.6. Inhibitor tolerance
1) Compounds originating in the biomass by hydrolysis. These include organic acids such as
acetic, glucuronic and galacturonic acids from the hemicellulose, and phenolic compounds
from the lignin. The inhibitoriest of these for both yeast and bacteria is acetic acid and
solubilized lignin.
2) Compounds formed by degradation of the products resulting from pretreatment and
hydrolysis of the biomass. Furfural from xylose and hydroxymethyl furfural from glucose lead
this group. It is completed by an assortment of aldehydes, acids and alcohols from lignin,
sugar and protein degradation.
3) Compounds from other sources. Metal ions resulting from equipment corrosion, sulfites,
sulfur dioxide and lactic acid introduced with other streams containing nutrients, cleaning
solutions and backset.
2.10. Previous study of bioethanol production
World fuel ethanol production
The commercial production of fuel ethanol relies mainly on the fermentation of sugar
and starch, while lignocellulosic ethanol entered the market only recently. USA and Brazil
have been the leading countries in the production of ethanol from corn starch and sugarcane
respectively, and the amount of ethanol produced by these two countries together in 2013
was 19,567 billion liters, accounting for 84% of the worlds production in that year. (Balat et al.,
2011).

Table 2.3: World fuel ethanol production by country or region (million Gallons)
Country 2007 2008 2009 2010 2011 2012 2013 2014 2015
USA 6,501 9,309 10,938 13,298 13,948 13,300 13,300 14,300 14,806
Brazil 5,019 6,472 6,578 6,922 5,573 5,577 6,267 6,190 7,093
Europe 570 734 1,040 1,290 1,168 1,179 1,371 1,445 1,387
China 486 502 542 542 555 555 696 635 813
Canada 211 238 291 357 462 449 523 510 436
Rest of 315 389 914 985 698 752 1,272 1,490 1,147
world
WORLD 13,123 17.644 20,303 23,311 22,404 21,812 23,42 24,570 25,682
Source: Renewable fuel association http://WWW.ethanolrfa.org/resources/industry/statics
Figure 2.7: Annual ethanol productions by the major producers, adapted from Mabee (2007)
Ethanol production in Ethiopia

Ethanol production in Ethiopia has been started since 1998/99 in Fincha sugar factory with the
capacity of 1,907 m3 per year. In 2010/11 Ethanol production in Metahra sugar factory with the
capacity of 6,373 m3per year. Currently, both factories produce 19,805m3ethanol per year
(Ethiopia Sugar Corporation). The main raw material for this product is molasses which is a by-
product of sugar production from the crystallization process. Now a day different researchers

conduct different research to produce bioethanol from different raw material such as fruit peel,
sugar beet, wood residue, agricultural residue, spent grain etc. all this works was economical
feasible but does not applicable in our country.
Table 2.4: Ethanol production in Ethiopia
Year Factory and Ethanol produced (M3)
Fincha Wenji Metahra Tendaho Total
2006/07 8,000 - - - 8,000
2007/08 8,448 - - - 8,448
2008/09 11,009 - 7,311 - 18,320
2009/10 13,834 - 11,700 9,602 35,136
2010/11 16,585 - 11,700 24,428 52,713
2011/12 18,515,000 15,428,000 28,183,000 38,205,000 100,331,000
2012/13 21,221,000 20,728,000 35,527,000 50,689,000 128,165,000
Source: East African Power Industry Convention 2007, Addis Ababa, Ethiopia

3. MATERIAL AND METHODS

3.1. Materials and equipment used for the experiments
Equipments
Oven-to dry the sample.
Crushers: - to crush the dried sample.
Sieves: - to sieve the crushed sample to the desired particle size.
Balances: - to weigh sample.
PH meter: - to measure the pH of the hydrolysate before fermentation.
Thermometer: - to control temperature of the sample under experiment (distillation).
Funnel and filter paper: - to separate the soluble liquid from non soluble part.
Graduated cylinders of different volumes: - for volume measurement.
Autoclave: - for sterilization and hydrolysis.
Pycnometer or Hydrometer: - measurement specific gravity.
Shaker:-to shake sample and its additives after hydrolysis and before fermentation.
Chemicals and material
 Barley spent grain: - Used as raw material.
 98% Sulfuric Acid (H2SO4):- used as a pretreatment and hydrolysis BSG.
 Yeast extracts (Agar):- used as media preparation.
 Sodium Hydroxide (NaOH):- used to adjust the pH of soluble cellulose and
hemicelluloses before fermentation.
 Urea: - used as media preparation.
 Dextrose sugar: - used as media preparation.
 Mg SO4.7 H2O: - used as media preparation
 Yeast (Saccharomyces cerevisiae):-used to facilitate the fermentation process.

3.2. Experimental set up
BSG Drying Milling
Hydrolysis Mixing Sieving
Filtration Yeast addition Fermentation Distillation
Figure 3.1: Laboratory experimental set up

3.3. Experimental procedure

3.3.1. Sample preparation
Barley Spent grain (BSG) is the major byproduct produced by the beer industry obtained from
dashine brewery factory Debre berhan, Ethiopia. The sample is transported by using plastic bag
to Debre brihan university institute of technology, department of chemical engineering
laboratory. The samples used for this study were prepared in chemical engineering laboratory.
1kg of Barley spent grain (BSG) is washed in order to remove unwanted matter and dried at
110oC for 20hour to achieve easily crushed material. Then the dried sample is milled and sieved
in to appropriate particle size which is less than 2mm. The milled sample was sterilized at 121oC
for 15 min and stored at room temperature.
3.3.2. Dilute acid pretreatment
The pretreated feedstock was mixed with 0.1ml H2SO4/g of dry matter sulfuric acid
solutions in 500-mL closed universal flasks with a liquid-to-solid(water to sample) ratio of 8
(g/g). The mixtures were allowed to stand for 10 min at room temperature in order to equilibrate
the acid concentrations between the bulk phase and the biomass. Hydrolysis was performed in an
autoclave at 121°C for 17 min.
Procedures 1: dilute sulfuric acid pretreatment
 Add 50 g of grinded dried barely spent grain in to 2000 ml conical flasks
 Add 400ml of distilled water and 5ml of sulfuric acid.
 The conical flasks capped with the help aluminum foul and allow the mixture for 10 min
at room temperature.
 Autoclave at a temperature of 121oC for 17minutes.
 After finishing the given pretreatment time and temperature the sample in autoclave and
allowed to cool and separate soluble from the non-soluble portion. The non-soluble
portion is hydrolyzed in the next steps and put the soluble solution in another 2000ml
conical flask.
Second stage diluted acid hydrolysis
The main purpose of this process is to degraded cellulose in to its monomer in the optimal
condition of temperature, acid concentration and reaction time. The procedure for this stage was
similar to first stage hydrolysis but energy required to degrade cellulose is higher than
hemicellulose so it needs higher temperature and concentration of acid.
Procedures 2: second stage dilute acid hydrolysis

The solid separated from first stage hydrolysis was washed and dried after drying the
Wight of sample become 42g.
Add 336ml of distilled water and 5.88ml of sulfuric acid to the non-soluble component
from drying steps.
The sample was then hydrolyzed in the autoclave with 140oC for 25min.
After hydrolysis separate the solid particle from liquid.
Then mix the soluble component with the previously filter solution from the pretreatment
step for the next procedure.
3.3.3. Filtration
The lignin and degraded cellulose which is called monomeric sugar was separated by using
filtration. Then the sugar solution or filtrate was neutralize and introduced into fermentation.
The lignin which obtained from this filtration process was dried and weight before use for
another purpose.
3.3.4. PH Adjustment
Before addition of any micro-organism to the above prepared samples, pH of these samples has
to be adjusted. Otherwise the micro-organism will die in hyper acidic or basic state. A pH of
around 4.5 -5.5 is maintained.
Mix pretreated and hydrolyzed solution, filtered, shaken substrate primarily checked for
PH using a digital pH meter. The pH then adjusted to 4.5-5.5.
Procedures in pH adjustment
Mixed samples were acid hydrolyzed, so it needs highly basic solution to bring the pH in
the range of 4.5-5.5.
Sodium hydroxide solution was added drop wise to the other flask with constant stirring
until the pH reaches to a range of 4.5-5.5.
If suppose the pH goes beyond 4.5-5.5, concentrated sulfuric or hydrochloric acid was
added drop wise to maintain the pH in the range.

3.3.5. Sterilization
The reactor and all the equipments that were used for fermentation purposes were sterilized
(Autoclaved). The sterilization was carried out at a temperature of 121oC for 15 minutes.
3.3.6. Fermentation
The fermentation process was carried out in incubator, at 30 °C for a 72h.The prepared
hydrolysate were adjusted to pH of 5.0 which is optimum for Saccharomyces cerevisiae
using 2M sodium hydroxide solution.
Media Preparation
The culture medium was prepared in 250 mL test tube by composed of
Yeast extract-----------10 g/l
Dextrose ---------------20 g/l
Urea --------------------5 g/l
Mg SO4.7 H 2 O--- 5 g/l

3.3.6.1 Procedures in Media Preparation
 The media was sterilized at 121oC for 15min and 0.50g of Saccharomyces cerevisiae
were added into 100ml prepared media at 250 mL conical flask.
 The conical flasks were properly covered with aluminum foil and placed to incubator for
24 h, at 30oC.
The Procedure for Fermentation
 The hydrolysate sample was conditioned at temperature of 30oC. This temperature is
favorable for the fermentation by which Saccharomyces cerevisiae.
 The pH of the sample was adjusted using 2M NaOH to make solution pH from (4.5-5.0)
to establish a favorable condition for S. cerevisiae.
 The hydrolysate sample was placed into incubator at 30oC and for 3 days.
 After 72 h fermentation, the samples were taken out and introduce into distillation to
separatethehydrousethano

3.3.7. Ethanol Separation (Distillation)

Distillation is the last step in the production of ethanol. It is a purifications step. Distillation is
the method used to separate two liquid based on their different boiling points. However, to
achieve high purification, several distillations are required. Separation was carried out by simple
distillation at 85oC for 2 hrs. All distillation experiments were carried out at a temperature of 85-
90oC and a distillation time of 2 hours.

STRG-101 / V-101
Distelled water storage tank SP-101
Flow Splitting
BSG
DDRNG-101 / DDR-101
Drum Drying
V-103
GRD-102 / GR-102 Dried BSG storage tank
Grinding
STRG-102 / V-104
RXN-101 / R-101
Dilute acid storage tank
Primary hydrolysis
PMP-101 / P-101
Pumping
PFLN-101 / PF-101
Plate & Frame Filtration
ST / V-105 RXN-102 / R-102

Hydrolyzate storage one Secondary hydrolysis
PMP-102 / P-102
DDRNG-102 / DDR-102
Pumping PFLN-102 / PF-102
Drum Drying
Plate & Frame Filtration
MXN-101 / MX-101
Mixing
STRG-103 / V-102
STRG-104 / V-106
Yeast
NEUTR-101 / NU-101
Neutralization
FRMN-101 / F-101
Fermentation
DSTL-101 / C-101 STRG-105 / V-107

Distillation Ethanol storage
FILL-101 / FR-101
Filling
Figure 3.2: Bioethanol production flow sheet from barley spent grain

4. RESULT AND DISCUSSION
4.1. Moisture content determination

Dried and clean petry dish weight (W1) is determined. About 100g, 125g, 150g of sample is
weighed in dray petry dish (W2) and dray at 110oc until constant weight is obtained and the
final dray sample is weighted (W3) again.Therfore the moisture content determined by
% moisture content = ∗ 100................................................................................... (4.1)
Table 4.1: Moisture content determination
Wight of Wight of Drying time(hour ) (W3) % of moisture

dish(w1) sample and content
dish(w2)
12 14 16 18 20
38.82 138.82 89.55 81.04 74.47 71.938 71.93 66.89
38.82 163.82 111.27 105.07 99.94 97.51 97.42 53.12
38.82 188.82 123.69 119.04 115.71 113.5 113.25 50.38
Average 56.8
moisture
4.2. Ethanol characterization

The PH and Specific Gravity of ethanol were determined after distillation and sugar
concentration after hydrolysis.
The sugar concentration was measured 10% brix are present in the sample after hydrolysis.
The pH of the sample before fermentation and after distillation was measured by using pH
meter and the result after distillation where 4.14 and before fermentation 4.5 to 5 to make
favorable condition for yeast growth.
4.2.1. Specific gravity of ethanol

The specific gravity of the ethanol was determined by using 20oC Pycnometer. The
25ml Pycnometer was cleaned and dried first and then weighed (W0), then after the
bottle was filled with ethanol, stopper inserted and reweighed to give (W1). The ethanol was

substituted with water after washing and drying the bottle and weighed to give (W2). The
expression for specific gravity (Sp.gr) is:
Sp.gr =
Where: W0- weight (g) of empty bottle
W1- weight (g) of bottle + sample
W- Weight (g) of bottle + water
Volume, density and alcohol concentration of ethanol
Table 4.2: Volume, density and alcohol concentration of ethanol
Volume of Density Ethanol
ethanol(ml) after Of ethanol(g/ml) Con. (w/w %)
distillation
33 0.9545 29.306
28 0.9363 39.52
34 0.9234 45.614
36 0.9189 48.3
27 0.962 24.78
30 0.95710 28
The experiment was conducted with constant hydrolysis acid concentration, hydrolysis time
and fermentation temperature but variable yeast concentration and fermentation time.
4.3. Effect of yeast concentration
The experiments was conducted with variables consisting of yeast amount (50, 100 and
150ml) and constant fermentation time.

Table 4.3: Effect of yeast concentration and the fermentation at 3 day
Attribute Yeast concentration (g)
50 100 150
Ml 28 36 34
Specific gravity 0.9363 0.9189 0.954
During the experiment 50ml, 100ml and 150ml of yeast were transferred in to 500ml of the
hydrolyzed liquor for fermentation for 3 days. After this the clear fermented was double
distilled and we get a clear color less liquor then calculate the specific gravity of the
liquor and read the alcohol content in volume percentage from standard table. In the above
table all the samples have around 35.92% alcohol content. The remaining part is water and
other mixtures that have near boiling point to them. But from these concentrations of yeast
the most the alcohol content is 100 ml which is 48.3%. The lower the yeast concentration
may not have enough power to consume all the glucose present in hydrolysis solution.
So that the quantity of glucose released as a waste was high. On the other hand using high
amount of yeast for the given media may compute to glucose present on the hydrolysis
solution for their growth. So that the final yield of alcohol can be reduce.
0.96
0.955
0.95
Specific gravity
0.945
0.94
0.935
0.93
0.925
0.92
0.915
0 20 40 60 80 100 120 140 160
Yeast concentration
Figure 4.1: Yeast concentration vs specific gravity

Table 4.4: Effect of Yeast concentration on alcohol content
Yeast 50 100 150

concentration(ml)
Alcohol content 39.52 48.3 45.614

(%)
60
50
Alcohol content
40
30
20
10
0
0 20 40 60 80 100 120 140 160
Yeast concentration
Figure 4.2: yeast concentration vs alcohol content
4.4. Effect of fermentation time

The experiments were conducted with variable fermentation time (3 day and 6 day) by
constant volume of acid, yeast concentration and temperature.
Table 4.5: effect of fermentation time on specific gravity
Yeast Fermentation time (day)

concentration
3 6
100 36ml 33ml
Specific gravity 0.9189 0.9545
The productivity of Bioethanol yield was observed to decrease with increasing fermentation
time beyond 3 days, probably due to microbial wastes such as CO2 that accumulates in the

fermentation media that tends to reduce the media pH thereby inhibiting microbial growth by
cyto-toxicity effect.
6
Fermentation time
0
0.915 0.92 0.925 0.93 0.935 0.94 0.945 0.95 0.955 0.96
Specific gravity
Figure 4.3: Specific gravity vs fermentation time
Table 4.6: effect of fermentation time on alcohol content
Fermentation time(day) 3 6
Alcohol content 48.3 29.306
60
50
Alcohol content
40
30
20
10
0
3 6
Fermentation time
Figure 4.4: Fermentation time vs Alcohol content

5. MATERIAL AND ENERGY BALANCE
5.1 Material balance

Laboratory scale mass balance
50 gram
Spent grain
5ml H2 SO4 400ml

1st stage hydrolysis
Water 336ml 2nd stage hydrolysis 5.88 ml H2SO4
Neutralizatio
NaOH n
Fermentation Yeast 100ml
10ml ethanol
Distillation
Figure 5.1: Laboratory scale material balance
Industrial scale mass balance

Table 5.1: Annual current and future demand of fuel ethanol in (litter)
Year Forecasted Existing local Supply gap
demand supply
2016/17 354,233,580 271,860,000 82,373,580
2017/18 461,566,354 271,860,000 189,706,354
2018/19 601,420,960 271,860,000 329,560,960
2019/20 783,651,511 271,860,000 511,791,511
2020/21 1,021,097,919 271,860,000 749,237,919
2021/22 1,330,490,919 271,860,000 1,058,630,588
2022/23 1,733,629,236 271,860,000 1,461,769,236

Source: - Survey of Large & Medium Scale and Electricity Industries, CSA and Customs
Authority
The supply gap in 2019/20 is 511,791,511 litters our production capacity is 400,000 then it
covers 0.08% of the gap.
Basis: one year/300 days
Production capacity: 400,000 litter per year of ethanol (99%) with plant operation of 300
days per year and the Plant attainment (i.e. the percentage of the available hours in a year)
was calculated as
% Plant attainment = ∗ 100 = ∗ 100= 83.3 %
In terms of mass basis
ƍ = /v.................................................................................................................................. (5.1)
m = ƍ x v =0.789kg/lit*400,000lit/yr = 315,600kg ethanol per year or 315.6ton/yr
Material balance for Molecular sieve

Molecular sieve used to increase the ethanol concentration from 95% to 99%.
Anhydrous Ethanol (ṀAE) =31.56 ton 31
Hydrous ethanol (ṀHE) =? Molecular XAE = 99%

sieve
XHE = 95% Xw = 1%
XW = 5% Water (W) =?
Xw=1, Xe = 0%
Where Xe= Percentage of Ethanol
Xw= Percentage of water
Over all mass balance
ṀHE=W + ṀAE................................................................................................................ (5.2)
Ethanol Component mass balance
ṀHE* XHE= ṀAE* XAE................................................................................................. (5.3)

From equation 5.3
Ṁ ∗ . ∗ .
ṀHE= = .
= 328.9 ton
The water that is trapped by the molecular sieve is calculated from equation 5.2
W= ṀHE - ṀAE
= 328.9 – 315.6 = 13.3 ton of water
Material balance on distillation
Distillation is used for recovery of ethanol from water-ethanol mixture. The mixture of
ethanol water which obtained from fermentation process was separated using two-stage
distillation column to recovered 95% ethanol from the mixture of 3.5% ethanol.
D1=328.9 ton
D=? XD1=0.95
2nd distillation
1st distillation
F XD = 0.5
ZF = 0.035
W1=?
Xw=1 , W=? Xw1 =0.99
Where
F=feed to the 1stdistillation
D=distillate from 1stdistillation
ZF=feed composition of light component
D1= distillate from 2nddistillation
W= bottom product
Over all mass balance on the 2nddistillation

D=D1+W1
D= 328.9 +W1................................................................................................. (5.4)
Ethanol Component mass balance on the 2nddistillation

0.5D=0.95D1 +0.01W1...................................................................................... (5.5)
Substitute equation 5.4 in to 5.5

0.5(328.9 +W1) = 0.95*328.9 +0.01W1
W1= 302.1 ton

D=315.6 + 302.1 = 617.7ton
Over all mass balance on the 1stdistillation
F = D+W
F= 617.7 +W.......................................................................................................... (5.6)

Ethanol Component mass balance on the first distillation
0.035 F = 0.5D +0................................................................................................... (5.7)
Substitute equation 5.6 in to 5.7

F= 8824.3 ton
W= F- 617.7 = 8206.6 ton
Material Balance on Fermentation
Material balance in the fermenter was carried out based on the fermentation reaction
C6H12O6 2C2H5OH +2CO2

180 g/ mol 92g/mol ethanol + 88 g/mol CO2 for 100% conversion.
Mass of filtrate (MF) =? Mass of co2 release (MCO2) =?
Fermentation
Mass of fermented mash
Mass of media (Mm) =? (MFm) =8824.3 ton

Assume 90% glucose can converted into ethanol if the fermentation process very effective
from literature.
. ∗ ∗ .
=
. ∗ . ∗
MG = . ∗
= 679.3tone
Where
D1 =hydrous ethanol
MG=mass of glucose
The amount of carbon dioxide produced is
∗ . ∗
Mco2= = =332 tone of CO2 released per year
Over all mass balance on the fermentation process
MF +MM =MFM +MCO2

MM =0.1MF
MF +0.1MF = 8824.3 ton +322 ton
MF = 8323.9 ton of filtered Hydrolyzates is required
Mm = 832.39 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) was 23%.
Filtration
Hydrolyzates (Mh) mass of filtrate (Mf) = 8323.9
ton
Xs=0.044 XL=1
XL=0.956 Lignin (Ml), Xs=0.77, XL=0.23 Xs=0
Mh=Ml +Mf...................................................................................................... (5.8)

Mass balance on the solid
Mh *0.044 =Ml *0.77
Mh= 17.5Ml....................................................................................................... (5.9)

Substitute equation (5.9) into equation (5.8) to calculate mass of lignin or total solid remain
after hydrolysis
17.5 Ml =Mf
Ml = (8323.9 ton)/17.5
= 475.7 ton of solid or lignin was generated per year.
The amount of hydrolysate is calculated from equation (5.8)
MH=8796 ton of hydrolyzates is need per year
Material balance in dilute acid treatment (Hydrolysis)
The mass balance was based on the laboratory result obtained; the liquid solid ratio was fixed
at 8:1, in the first stage hydrolysis 1.23%w/w acid concentration was used to decompose
hemicellulose and in 2ndstage 1.73 %w/w acid concentration was utilized to decomposed
cellulose in to its monomer which is called glucose. The amount of solid charged in to the
2ndstage was 60% of the total solid charged into the 1ststage and the, remain 40% was
decompose in the 1ststage 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
to Dehnavi, (2009) 27% of barley spent grain is cellulose.
MA1 (1.23%)
M BSG1 1st stage
MC =0.27(MBSG) MH =8796 ton/year
0.60MBSG1 MG= 679.3 tone
MA2 (1.72%) 2nd stage
By using stoichiometry equation the required dry barley spent grain (M BSG) is calculated as
. ∗ . ∗
= =
0.27*0.95* MBSG= (162* 679.3)/180

M BSG1=2383.5 ton of dry barley spent grain is required per year.
Mass balance on the 1st stage hydrolysis

The liquid solid ratio in the 1ststage was (8:1)
MA1=8*2383.5
MA1= 19,068 ton per year of 1.23% diluted sulfuric acid is required per year in the 1ststage
hydrolysis. The acid required in this stage is 234.5 ton/year.
The same is true for the 2nd stage hydrolysis; liquid solid ratio was (8:1)
MA2=8*0.60MBSG
MA2= 11,440.8 ton per year of 1.72% diluted sulfuric acid is required per year in the
2ndstage hydrolysis. The acid required in this stage is 196.8 ton/year.
Mass balance on dryer
MWBSG Dryer MBSG = 2383.5 ton
Xw=0.493 Xw=0.1
Xs=0.507 MW Xs=0.9
MWBSG=Mw + MBSG.............................................................................................. (5.10)
Component mass balance on the solid BSG
0.507 MWBSG=0.9MBSG
MWBSG= (0.9*2383.5)/0.507
= 4231.1 ton of wet BSG is required
Mw = MWBSG – MBSG = 1397.6 ton of water
5.2. Energy balance

Energy balance on dryer
The energy balance was performing based on the laboratory results.The basic of the
calculation is one hour.

T=25oC Dryer T=105 oC
M= 4231.1 ton
The required energy is calculated as
Q = m Cp ∆T................................................................................................................ (5.11)
Where
Q= required energy
M=mass of wet BSG
Cp=specific heat of wet BSG, Cps =0.44 CPw
Cp = CPw*Xw + Cps *Xs
= 4.18*0.493 +4.18*0.44*0.507
= 3 kJ/kg k
Q = m Cp ∆T
= 4231.1 *3 (105 - 25)
Q= 1,015,464 kJ/year = 3,384.88kJ/day
Energy balance on hydrolysis
The energy used for hydrolysis process is generated from water steam.
MA1= 2648.3 kg/hr
MBSG1=331 kg/hr 1st stage T1=121o C
T=25oC
MBSG2= 198.6kg/hr MH=122.2kg/hr
MA2=1,589kg/hr 2nd stage T2=140oC
T=25oC
Where
MBSG1=mass of BSG input to 1st hydrolysis stage
MBSG2=mass of BSG input to 2ndhydrolysis stage
MA1=mass of water and acid solution input to 1ststage hydrolysis
MA2= mass of water and acid solution input to 2ndstage hydrolysis
The amount of heat required for the hydrolysis process

Q = M1Cp1ΔT1 + M2Cp2ΔT2.................................................................................... (5.12)
∗ . . / ∗ .
CP1= = 4.136kJ/kg k
. /
.
∗ . , / ∗ .
CP2 = . =4.142 kJ/kg k
, /
Q=2979.3*4.136(121-25) + 1,787.6*4.142(140-25)
= 2,034,436.44KJ/hr
Energy balance on fermentation
Mm=115.6 kg/hr
MF=1156 kg/hr Fermentation MFm=1225.6kg/hr
MCO2=46.11 kg/hr
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 CO2at 30oC = 0.846kJ/kg k
QMIX = QCO2 + QFh+ Q.............................................................................................. (5.13)
Q = QMIX - QCO2- QFM
Q= Mmix Cpmix ΔT-MCO2
CpCO2 ΔT-MFmCpFm ΔT
CpFm=Cpmix*Xmix+CpCO2*XCO2
XCO2=4.611 / (115.6 +11.56) = 0.0363
XFm=1-0.0363=0.9637
CpFm= 4.14*0.9637 + 0.846*0.0363
= 4.0204kJ/kg k
Q=1271.6*4.14*(30-25) – 46.11*0.846*(30-25) – 1225.6*4.020(30-25)
= 1492.52kJ/h

Energy balance on distillation boundary
F=1225.6 kg/hr D=85.8 kg/hr
XFW=0.965 Distillat Xde=0.95

ion
XFE=0.035 Xd=0.05, T=25oC
T=30oC
W=1140 kg/hr, Xw=1, Xe=0
Specific heat capacity of distillate (Cpd)

Cp = 0.95*2.43(distillate cp)+0.05* 4.18= 2.518kJ/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*85.8kg/hr = 214.5 kg/hr
V=L+D=214.5 kg/hr + 85.8 kg/hr = 300.3kg/hr
From equilibrium data, boiling point of ethanol at 96% is 78.1oC
At steady state condition
Input=output
HF = HD + HL +QC......................................................................................................... (5.14)
QC=HF-HD-HL
Assume complete condensation, therefore
Enthalpy of Vapor = Latent + Sensible heat
HV = mvλv + mvcpΔT............................................................................................ (5.15)
=300.3kg/hr* 789+300.3kg/hr* 2.518kJ/kg k (78.1-25) =277,088.55 kJ/hr
Overall balance to determine QB
Input = Out put
QB + HF = QC + HD + HW...................................................................................... (5.16)
QB=QC+HD+HW-HF
HF = mFCpΔT
Cp=0.035*2.43+0.965*4.18 = 4.118 kJ/kg k
HF=1225.6 kg/hr*4.118*(30-25) = 25,235.1 kJ/hr
HW = MwCpΔT=1140 *4.18 (100-25) =357,390 kJ/hr
QB = QC + HD + HW - HF
QB= 277,088.55 kJ/hr + 0 + 357,390 kJ/hr -25,235.1 kJ/hr = 609,243.45 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.................................................................................................................... (5.17)
Ms=Q/ λs = (609,243.45kJ/hr) / (2174 kJ/kg) = 280.24 kg/hr
Mass of water that is able to condense is removed with temperature rise of 30oC
, . /
QC=Mw Cp ΔT, M = Δ
= . ∗
= 3,668.1KJ/kg
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 ethanol and
raise the concentration of ethanol to around 99.9%.
Basic; one hour Anhydrous Ethanol (ṀAE) =43.8 kg/hr
Xw=1%, Xe= 99%
Hydrous ethanol (ṀHE) = 45.7 kg/hr Molecular T=95oc

Sieve
XHE=95%
Xw=5%
To=25oC
Mass of water =1.85 kg/hr
Xw=1, Xe=0
Cp is calculated based on their percentage of the input mass of hydrous ethanol

Cp = 0.95*2.43+0.05*4.18 = 2.643kJ/kg k
The temperature entering the molecular sieve is 25C and exit is at a temperature of 95oC
Q=M Cp ΔT = 45.7* 2.643*(95-25) =8,455 kJ

6. SIZING AND DESIGN OF MAJOR EQUIPMENTS

6.1. Sizing
Assumption:
All tanks are 90% full and 10% safety factor.
Sizing of equipment is depending on the material balance calculated on the above section.
Basis: one day
Sizing of dried spent grain storage tank
Table 6.1: Specification of dried spent grain storage tank
Material of construction carbon steel
Material handled dried spent grain
Density of spent grain 0.336g/cm3=336kg/m3
Density = mass/volume
The required volume of storage tank is for 2 days
/
Volume dried spent grain = = /
= 22.3m3/day
.
V= .
= 24.8m3
Sizing of water storage tank

Table 6.2 Specification of water storage tank
Material of construction carbon steel
Material handled dried spent grain
Density of spent grain 0.336g/cm3=336kg/m3
Mass of water = 10,733.17kg/day

, . /
Vof water tank = /
=107.3m3
.
V= .
= 115.2m3

Sizing of hydrolysis reactor

Table 6.3: Specification of hydrolysis reactor
Materials of construction Stainless steel
Material handled slurry of BSG and water mixture

3
Density of slurry 888 g/
Mass of slurry = 29,328 kg/day

, /
V= = /
=33.03m3
.
Vvessel = .
= 36.7 m3
Sizing /design of agitator in the hydrolysis reactor

Assume the length to diameter ratio. I.e. L/d= 1.5, L=1.5 D
Volume of the tank/mixer =1.6m3
V= ∗
∗ .
1.6m3 =
D =1.4 m, L =2m
Diameter of agitator = 0.33*D=0.46m
 Liquid to be handled -------------------------------------------------- dilute sulfuric acid
 Density of liquid ----------------------------------------------------- 990.018
 Temperature of liquids ------------------------------------------------- 25 ℃

 Material of construction ------------------------------------------------ stainless steel
 Mass flow rate of liquid ------------------------------------------------ 1560

/
V= = . /
= 1.58 m3/day
.
v= .
= 1.76m3/day

Sizing of hydrolyzates storage tank

Table 6.4: Specification of hydrolyzates storage tank
Materials of construction rubber-linked cs
Material handled hydrolyzates of BSG

3
Density of liquid 880 g/
Temperature of the slurry 20oC
Mass of slurry = 29,328 kg/day

, /
V= = /
= 33.33m3/day
. /
V= .
= 37 m3
Sizing of Fermenter
Table 6.5: Specification of fermenter
Material construction carbon steel
Material to be handled Hydrolyzate
Shape dome shaped

3
Density of liquid 880 /
Temperature 30oc
Mass of feed = 30518.4 kg/day

. /
V= = /
= 34.68m3/day
.
V= .
= .
= 38.53m3
Sizing of ethanol Storage tank

Table 6.6: Specification of ethanol storage tank
Material of constriction Carbon steel
Material handled Ethanol
Density 0.789kg/lit=789kg/m3
Temperature of liquid 20oc

Mass of ethanol per day is = 1,051.2 kg/day

, . /
V= = /
= 1.33m3/day
.
V vessel = .
= 1.5m3
Pump for delivering of water
Type..................................................................................... centrifuge pump

Head.............................................................................................. 4m
Slurry to be handled ......................................................................distilled water
Density .........................................................................................1000 g /m3
Temperature of liquid....................................................................25oC
Materials of construction ............................................................carbon steel
3
Capacity.......................................................................................0.04 /s
Pump for delivering filtered hydrolyzates
Type .................................................................................centrifuge pump
Head......................................................................................4m
Slurry to be handled..............................................................hydrolyzate
Density..................................................................................888 /m3
Temperature of liquid ..........................................................25oC
Materials of construction................................................... carbon steel
3
Capacity.......................................................................... 0.004 /s
Pump for delivering hydrolyzate

Type ...............................................................................diaphragm
Head................................................................................. 4m
Slurry to be handled ...........................................................hydrolyzate
Density................................................................................880 /m3
Temperature of liquid....................................................... 25oC
Materials of construction................................................ carbon steel
3
Capacity.........................................................................0.025 /s

6.2. Design of process equipment

Dried barley spent grain storage tank
Assuming
Operating pressure .........................................................................1 atmosphere = 101.3 kpa
Operating temperature ....................................................................25 ℃
Assuming the storage tank is a cylindrical
V cylinder = 24.8 m3
V=
H/D = 2, H = 2D
( ∗ )
52.7=
∗ ∗ .
D= ∗
= ∗ .
= 2.5m
H = 2* 2.5 = 5m
Angle of the cone
Let a = 0.1 ∗ H = 0.5m
r= = 1.25
Using Pythagoras theorem

a2+ r2= S2
S2= (0.5)2+ (1.25)2= 1.8 m2 and S = 1.34 m
And to find the value of the angle
.
cos α = = .
= 0.37
α = cos−1 (0.37) = 68.260

(Angle of the cone) = 2 ∗ α = 136.56o
Wall thickness of the dry barley spent grain storage tank (vessel)
There will be a minimum wall thickness required to ensure that any vessel is sufficiently rigid
to withstand its own weight, and any incidental loads.
a) Thickness due to the pressure loading
∗
t=
∗ ∗
(Working pressure)= Patm+ ρ ∗ g ∗ H = 101.3 kpa + (336kg/m3*9.81 m/s*5 m) = 117.78Kpa

Internal pressure= safety factor +working pressure= 0.1*117.78+ 117.78= 129.56 Kpa =
0.129 N/mm2

∗ . ∗
t= ∗ ∗
= = 1.4 mm add corrosion allowance 2mm
∗ ∗ . .
t=1.4+2=3.4mm.
From the standard for 3 to 3.5mm diameter the minimum thickness is 12 mm and considering
the corrosion allowance.
Design of head and closure of cylinder
Choice of closure----------Flat ends (with a full face gasket
Justification
1. Why flat end
• Applicable for low pressure
• Fabrication cost is low
The pressure exerted on the flat ends of storage tank is only atmospheric pressure
that is: Patm=101.3.
Therefore: - design pressure can be calculated by using safety factor 10%.
P( )= 0.1∗ + =111.43 =0.11 / 2
t= ∗ √
Where: - C = design constant dependent on the edge of constraint = 0.4

Design stress “f” for carbon steel at room temperature is 135N/mm2 and the Diameter is 1.5m
.
t=0.4 ∗ 2500√ = 28.54 mm
Corrosion allowance 2mm the thickness of head became 30.54mm

Design of conical section and end closure
Justification
1. Why conical section
• Facillating the smooth flow
• Applicable for removals of solids from the process equipment
• Resistance to high stress
• Decrease surface area
The internal or design pressure can be calculated as the sum of pressure due to gravity and
pressure due to the atmosphere outside the tank.
Assume volume of cone is 5-10% of the total volume for this taking the average 7.5%.
VC=0.075*24.8=1.86m3
Diameter of cone: Dc= ID
∗
t= ∗ = 3.78mm
∗ ∗ ∝

WEIGHT OF STORAGE D
Dm=ID + t
Dm=2500+1.4=2.5m
=240∗ ∗ ( +0.8∗ )t
Where Wcct=weight of storage tank
Dm =mean diameter α a
t=thickness
CV= design factor to account for the weight =1.08 Figure: BSG storage tank
H = height of storage
Wft=240*1.08*2.5(5+0.8*2.5)2.5
=11,340 N
Wind loading
Fw=PD*Dm where Fw=wind loading
=1280*2.5=3,200N/m PD=dynamic pressure (a wind speed of 160km/hr. can
be used for preliminary design studies equivalent to wind pressure 1280N/m2
Analysis of stress
σw = /I( /2+ t ) where Mc =bending moment
Mc= /2*H2
=3,200/2*(52)
=40,000
( )
I= = Where I= the moment area
OD=ID+2t
OD=2.5+2*0.0014) =2.503m
I=9.216*109mm
σw =13.47*10-9N/mm2
Support leg (skirt support)
Justification
1. Why skirt support
• Skirt supports are recommended for vertical conical cylindrical vessels as they do not
impose concentrated loads on the vessel shell; they are particularly suitable for use with tall
columns subject to wind loading.
Assumption
Height of the leg--------------------------0.5m

Thickness of leg---------------------------17mm
Inner diameter ......................................700mm
Bending moment (m) =wind load∗H2/2
=1608.96*(0.5)2 /2=201.12N
∗ ∗
Bending stress ( σb) =( )π ∗
=0.03N/mm
Dead weight stress

σs = / ( + )
,
= . ( )
=0.296N/mm2
Tensile stress=bending stress + weight stress

σ c=σb+ σs
=0.03+0.06=0.09N/mm2
Comprehensive stress= bending stress-weight stress
=0.03-0.06=-0.03N/mm2
Design criteria.
σcompress≤0.125 *E*( / ) sin ө (correct),
σ tensile≤135Jsin (correct) so the criteria is satisfied
Where E=young's modules (200,000)
α=90°
J=0.85
Design of batch fermenter
Assuming
Material of construction --------------------------------------------------Stainless Steel
Volume of tank ------------------------------------------------------------38.53m3
f = design stress of the type of material construction -----------135N/mm2
j = welded joint efficiency (double welded butt or equivalent) = 0.85 D
Operating pressure = 1 atmosphere -------------------------------- 101.3 kpa
Operating temperature ------------------------------------------------30°C
Assuming the fermenter is a cylindrical tank
Justification
Why batch fermenter
α a
•Reduced risk of contamination due to short period
•Low capital investment
•More flexibility with varying product Figure batch fermenter
•Small capacity
Since the shape of the tank is conical cylindrical and taking height to diameter ratio 2m
π
V= *h
h/D =2, h=2D

.
38.53== ∗2
D=4.95m
h=9.9m
Wall thickness of the fermenter tank
There will be a minimum wall thickness required to ensure that any vessel is sufficiently rigid
to withstand its own weight, and any incidental loads
t=pi*Di/2fj-pi
Working pressure=patm +pgh
=101.34 + (880*9.81*9.9) = 186.8N/m
Safety factor 10%
Internal pressure=0.1*186.8+186.8=205.48N/m=0.205N/mm
∗ . ∗
t= ∗ ∗
= ∗ ∗ . .
= 4mm
Thickness due to Wight of liquid

tc = ρ *h*g*D/2fj*1000=2.13mm
Total thickness of the tank
t total=t+tc=4+2.13=6.13mm
Add Corrosion allowance=2mm = 8.13
Design of head and closure (ellipsoidal head)
∗
t= ∗ ∗ .
Pressure exerted on the top of the tank is only atmospheric pressure so that patm=101.3kpa
Safety factor 10%
101.3*0.1+101.3=111.43kpa=0.11143N/mm2
t=0.64mm
Add corrosion allowance 2mm
2+0.64=2.64mm
But the minimum thickness for vessel diameter 1 to 2 meter is 7mm considering corrosion
allowance

Design of conical section

Thickness of conical head
Let a=0.1h then a=0.1*9.9=0.99m, r=D/2 = 2.47m
Using Pythagoras theorem
s2=r2+a2
=2.472+ 0.992
s=2.66m
Cos α =a/s= 0.372
α =68.20
Assume Diameter of cone: Dc=ID
∗
t= ∗∗
* α
= 10.75 mm
Weight of fermenter
Dm=ID+ t=4.954m
Wft=240*Cv *Dm (h+0.8*Dm)*t=75,820.83N
Where:
CV=a factor to account for the weight of nozzles, man ways, internal supports etc. which can
be taken as:
1.08 –for vessels with only a few internal fittings
1.15 –for distillation columns or similar vessels with several man ways and with plate support
rings or equivalent fittings
HV=height of the cylindrical section, m
t=wall thickness, mm from standard depending on the diameter of the tank= 7 mm
Dm=mean diameter of vessel or tank
Wind loading
Fw=PD *Dm= 6,341.12N/m
Where FW=wind loading
PD=dynamic pressure
Analysis of stress
∗( )
Stress =
OD=ID+2t=4.954m
I= /64(OD4-ID4) = 9.53*10 10mm4
Mc=Fw/2*h2=310,746.6Nm

Stress=8.08*10-12N/mm2
Support leg (skirt support)
Mild steel legs, diagonal cross bracing and center supports are used in place of bases when
applications require
Assume
The height of leg= 1m
Assume thickness of leg= 15 mm
Wind load is 640N/m
Bending moment (m) =wind load*leg height/2
= (640∗1 )/2 Nm=320Nm
Bending stress =4∗ ∗1000/( + ) ∗
σb = 4*320*1000/(250+15)*3.14*15*250
=0.41N/mm2
σx= ( )
=11,414.915/(3.14*(250+15)*15)
=0.9145N/mm2
Tensile stress=bending stress + weight stress
σc =σb + σx
=0.41+0.9145=0.375N/mm2
Comprehensive stress=bending stress-weight stress
=0.41-0.9145=-0.5045N/mm2
Design criteria
σ compress ≤ 0.125 *E*( )sinө , criteria are satisfied.
Where E=young's modules (200,000)
α=90°
J=0.85
σ tensile≤135Jsin criteria are satisfied

Design of Distillation Column

Specification
Mode of operation Continuous
Degree of separation 95% ethanol and %5 water @ the distillate
Condenser type total condenser
Contacting device plate type
Reboiler type partial Reboiler
Operating pressure 0.5 bar
Vapor velocity 0.75 m/s
A continuous fractionating column is to design to separate 1225.6kg/hr of ethanol water

mixture with 3.5% ethanol and 96.5% water so as to give 95% ethanol and a waste of 100%
water. A reflux ratio of 2.5 of product is assumed to be used. The column works at a vacuum
of 0.4bar with a vapor velocity of 0.4 - 0.6 m/s and takes 0.5 m/s (J and J NewYork 2005).
Molecular weight of ethanol = C2H5OH = 2*12 + 5 + 16 +1 = 46gm/mol
Molecular weight of water = H2O = 2 + 16 = 18gm/mol
Boiling point of ethanol = 78.130C = 351.13k
Boiling point of water = 1000C = 373k
Feed mean molecular weight = 0.035*46 + 0.965*18 = 18.98gm/mol
Specific heat of feed = 0.035*2.72 +0.965*4.18 = 4.98kJ/kg.k
Water mole fraction on feed
. /
YF = . . = 0.986=98.6%
Water mole fraction in distillate

/
YD = =0.1186=11.86%
Water mole fraction at the bottom

. /
YB = . = 0.996=99.6%
Ethanol mole fraction in the feed

. /
XF = . . =0.01398=1.4%
Ethanol mole fraction in distillate

/
XD = =0.8814=88.14%

Ethanol mole fraction in bottom

XB = 1%
Reflux ratio = 2.5
Boiling point (average) of feed is:-∑ XiTi = 0.01398*350C + (1-0.01398)*350C = 350C =
308K.
Molar latent heat of mixture in feed
λ = 0.035 ∗ 790 + 0.965 ∗ 2376.7 = 2,321.165KJ/kg
Specific heat capacity of the feed
CPf= 0.035*2.72+0.965*4.18=4.13KJ/Kg.k
Calculate the value of the ratio q given by
( ∗ )
q= =
latent
( )∗ . , .
Heat to vaporize one mole of feed (q) = , .
=1
The slope of the q-line is = =∞

.
The slope of the operating line for the rectifying section is = = .
=0.714
.
Y-intercept of operating line for the rectifying section is= = .
=0.25
X(liq) 0.0190 0.0721 0.0966 0.1238 0.1661 0.2337 0.2608 0.3273
Y(vap) 0.1700 0.3891 0.4375 0.4704 0.5089 0.5445 0.5580 0.582
X(liq) 0.3965 0.5079 0.5198 0.5732 0.6763 0.7472 0.8943
Y(vap) 0.6122 0.6564 0.6599 0.6841 0.7338 0.7815 0.8943

1.2
0.8
0.6
Y
Series1
0.4
45 degree line
0.2
0
0 0.2 0.4 0.6 0.8 1 1.2
X
Figure 6.1: McCabe-Thiele diagrams to determine number of stage
From the above information, number of theoretical plate is 9 and feed is introduced at the 6
stages.
Diameter at the top
The principal factor that determines the column diameter is the vapor flow-rate. The vapor
velocity must be below that which would cause excessive liquid entrainment or a high-
pressure drop.
Temperature at the top is 85 0 C (358 k) and ideal gas behavior.
V = Where n is molar flow rate of vapor n=30.03kg/hr
R= 0.082atm.m3/kmol.k
P= 0.4atm
30.03 ∗ 0.95 30.03 ∗ 0.05
n= +
46 18
kmol
n = 0.62 + 1.67 = 2.288
hr
. ∗ . . . ∗
V= = .
=168
Cross-sectional area
V 168m3/hr
A = = = 0.0933m
linear velocity 0.5 m ∗ 3600s
s hr

πD
A =
4
∗ .
D= = .
= 0.34m
Diameter at the bottom

Temperature of the bottom is 1000C (373k).
nRT M
V = and n =
P M
atm. m3
R = 0.082 , P = 0.4 atm and T = 373 K
Kmol. k
kg
114
n= hr = 6.33 kmol
kg hr
18
kmol
nRT 6.33 ∗ 0.082 ∗ 373 m
V= = = 484.278
P 0.4 hr
Cross-sectional area of the bottom column
V 484.278 m3/hr
A = = = 0.27 m
linear velocity 0.5m ∗ 3600s/hr
s
The diameter of the bottom column
4 4 ∗ 0.27
D= = = 0.6m.
3.14
We are selecting sieve or tray type of plate by considering the following criteria:-
It provides good vapor-liquid contact because the liquid rely for some time on the plate by the
vapor. It is the cheapest and satisfactory for most application.

7. FINANCIAL EVALUATION
7.1. Cost estimation
Economic Evaluation of the Project
Plant parameters
Capacity= 400,000 litter per year
Number of shifts per day = three
Working Days per year = 300
Purchased equipment cost
Table 7.1: Equipment cost estimation
Number Equipment Size of Material of Quantity Cost of equipment ETB

name equipment construction
Unit cost Total cost
1. Distillation 9 stage& 6m Stainless 2 397,917 647,834

height steel
2. Fermenter 38.53m3/3day Stainless 1 80,764 80,764

steel
3. Propagation 1m3 Stainless 1 15,000 15,000

tank steel
4. Drier 11 m2 Carbon 1 115,328.6 189,328.6

steel
5. Water 57.6m3/day Stainless 2 35,819.3 71,638.6

storage tank steel
6. Dilute acid 3.63m3/day Carbon 1 52,473.3 52,473.3

treatment steel
tank
7. Centrifugal 500-1000 Carbon 1 44,674.74 44,674.74

separation rpm steel
8. Shell and 2.2 m2 Stainless 1 40,740 40,740

tube HE steel

9. Plate heat 17.5m2 Stainless 1 40,153 40,153

exchanger steel
10. Electrical 14.13m2 Stainless 1 106,600 106,600

boiler steel
11. Condenser 1m2 Stainless 1 35,920 35,920

steel
12. Molecular 4.57kg/hr Stainless 1 31,919 31,919

sieve steel
13. Grinder kg Carbon 1 13,087.9 13,087.9

33.1
hr steel
14. Hydrolysis 37m3 Carbon 1 58,092 58,092

steel
15. Dry spent 24.8m3 Carbon 1 56,555.37 56,555.37

grain steel
storage tank
16. Ethanol 1.5m3 Stainless 1 11,284.57 11,284.57

storage tank steel
17. Sulfuric 1.76m3 Stainless 1 11,409.3 11,409.3

acid storage steel
tank
18. Pump 1 2.2 gpm Carbon 1 4,540 4,540

steel
19. Pump 2 0.5 gpm Carbon 1 2,310 2,310

steel
20. Pump 3 2 gpm Carbon 1 3,125 3,125

steel
Total purchased equipment cost 1,517,449.38

Table 7.2: Estimation of fixed capital investment
Direct cost (DC) Percentage (%) Price ETB
Purchased equipment 100%E 1,517,449.38
Purchased equipment 25%E 379,362.35

installation
Instrumentation and 10%E 151,744.94

control
Piping (installed) 15%E 227,617.4
Electrical( installed) 12%E 182,094
Building (including 20%E 303,489.9

service)
yard improvement 41%E 622,154.25

Service facilities
Land 5%E 75,872.47
A. Total direct cost (TDC) 3,459,784.7
Indirect cost (IC)
Engineering and 7%DC 242,184.93

Supervision
Construction expenses 5%DC 172,989.23

Contractor’s fee
Contingency 5%E 75,872.47
B. Total indirect cost 491,046.63
Fixed-capital investment (A+B) 3,950,831.32
Working capital (WC) = 15% TCI
Total capital investment = FCI + WC

TCI= FCI/ 0.85= 4,648,036.85
Then WC = 0.15* TCI = 697,205.53
A. Manufacturing cost = DPC+ FC + POC
a. Direct production cost (DPC)

1. Raw material
Table 7.3: Raw material
Item Quantity (tone/year) Unit price (birr) Total cost(birr)
Barley spent grain 2,383.5 150 357,525
Sulfuric acid 368.5 4,000 1,474,000
Sodium hydroxide 25 6,000 150,000
TOTAL 1,981,525
2. Operating labor
Table 7.4: Operating labor
Human resource No Monthly Salary per Annual salary

person
Manager 1 12,000 144,000
Professional 6 9,000 648,000
Semi- professional 6 7,000 504,000
Non professional 10 2,500 300,000
Total 1,596,000
2. Direct supervisor and clerical labor = 12% of operating labor = 191,520

3. Utilities (15% of TPC) = 0.15 TPC = 487,775.7
4. Maintenance and repairs =6% of FCI = 237,049.88
5. Operating supplies = 0.8% of FCI = 31,606.65
6. Laboratory charge= 12% of operating labor = 191,520

7. Patent and royalties = 3%of TPC= 97,555.14

DPC=1+2+3+4+5+6+7+8=4,642,552.4
b. Fixed charge(FC)
1. Deprecation(D)= 10%of FCI = 395,083.13
2. Insurance (I) = 0.7%FCI= 27,655.82
Then: FC=D+I= 422,738.95
FC= 13%TPC
TPC= 3,251,838.071
c. Plant overhead cost (POC) = 10%TPC=325,183.8
Manufacturing cost =DPC+FC+POC= 5,390,475.13
B. General expanse=AC+DSC+RDC
a. Administrative cost (AC)=4%TPC= 130,073.52
b. Distribution and selling cost(DSC)=9%TPC=292,665.43
c. Research and development cost(RDC)=5%TPC= 162,591.9
d. Financing (interest)=8 %TC= 371,842.95
General expanse= a+b+c = 585,330.85
C. Total product cost (TPC)=manufacturing cost +general expense = 5,975,806
Selling price
Table 7.5: selling price determination
Product Qty(l) Selling Price/l Total Price(birr)
Ethanol 400,000 19 7,600,000
7.2. Feasibility Study (economics)
The feasibility study for a chemical process design investigates both the technical and
economic feasibility of the proposed project. This feasibility study is only an introductory
assessment, at this stage the process route has not yet been finalized although a preferred
route may be apparent. Part of the work in preparing the feasibility study is to obtain
information regarding the alternative process routes, and to provide an assessment of the
suitability of these routes for the particular project.

Table 7.6: Determination of payback period
Year 0 1 2 3 4
- - - -901,376.41 405,598.2
4,648,036.85 3,428,193.99 2,179,307.19
- - -1,248,886.8 - 1,306,974.6
1,219,842.86 1,277,930.78
A. Rate of return
a. Before tax
, , .
ROR = * 100% = , , .
* 100% = 27.5%
Net profit after sell tax (35%) = 830,655

b. After tax
,
ROR = * 100% = , , .
*100% = 17. 9%
,
Turnover ratio = = , , .
= 0.21
, .
Payback period = 3+ , . .
= 3.7 Years
Calculation of present value, net present value and net present value ratio
PV = 9,380,695.1
NPV=PV+0 year including = 4,732,658.25
NPVR=NPV/NPV (initial) =1.02
Break-even analysis
The break-even analysis establishes a relationship between operation costs and revenues. It
indicates the level at which costs and revenue are in equilibrium. To this end, the break-even
point of the project including cost of finance when it starts to operate at full capacity is
estimated by using income statement projection.
Variable cot = raw material cost + utility cost =2,469,300.7
Fixed cost = TPC – Vc= 3,506,505.3
, , .
Breakeven point = ( )
*100 = , , , , .
= 0.683%
Break-even volume =0.683*7,600,000 = 5,194,115

The project cash flow diagram
Therefore: - the project is visible b/c
1) NPV>0
2) Payback period =3.7 year
3) NPVR>1

Table 7.7: cash flow
Year 0 1 2 3 4 5 6 7 8 9
Capacity utilization - 84% 86% 88% 90% 92% 94% 96% 98% 100
A. Cash in flow 6,384,000 6,536,000 6,688,000 6,840,000 6,992,000 7,144,000 7,296,000 7,448,000 7,600,000
Revenue 6,384,000 6,536,000 6,688,000 6,840,000 6,992,000 7,144,000 7,296,000 7,448,000 7,600,000
B. cash out flow -4,648,036.85 5,164,157.14 5,287,113.2 5,410,069.22 5,533,025.4 5,655,981.58 5,778,937.69 5,901,893.81 6,024,849.95 6,147,805.97
Investment -4,648,036.85 - - - - - - - - -
Raw material and other 1,664,481 1,704,111.5 1,743,742 1,783,372.5 1,823,003 1,862,633.5 1,902,264 1,941,894.5 1,981,525
Operating labor 1,340,640 1,372,560 1,404,480 1,436,400 1,468,320 1,500,240 1,532,160 1,564,080 1,596,000
Direct supervision 160,876.8 164,707.2 168,537.6 172,368 176,198.4 180,028.8 183,859.2 187,689.6 191,520
Utility 409,731.6 419,487.1 429,242.6 438,998.1 448,753.64 458,509.16 468,264.67 478,020.19 487,775.7
Maintenance and repairmen 199,122 203,863 208,603.9 213,344.9 218,085.89 222,826.89 227,567.89 232,308.89 237,049.88
Operating supply 26,549.6 27,181.7 27,813.85 28,446 29,078.12 29,710.25 30,342.38 30,974.52 31,606.65
Laboratory charges 160,876.8 164,707.2 168,537.6 172,368 176,198.4 180,028.8 183,859.2 187,689.6 191,520
Patent and royalty 81,946.32 83,897.4 85,848.52 87,799.62 89,750.73 91,701.83 93,652.93 95,604.04 97,555.14
Fixed charge 355,100.72 363,555.5 372,010.3 380,465.1 388,919.88 397,374.66 405,829.44 414,284.22 422,738.95
Plant overhead 273,154.4 279,658.1 286,161.7 292,665.4 299,169.1 305,672.77 312,176.45 318,680.12 325,183.8
General expense 491,677.9 503,384.5 515,091.15 526,797.8 538,504.42 550,211.03 561,917.65 573,624.27 585,330.85
Gross profit -4,648,036.85 1,219,842.86 1,248,886.8 1,277,930.78 1,306,974.6 1,336,018.42 1,365,062.31 1,394,106.19 1,423,150.05 1,452,194.03

Table 7.8: determination of present value

Year 0 1 2 3 4 5 6 7 8 9
Cash - 1,219,842 1,248,88 1,277,930 1,306,97 1,336,018 1,365,062. 1,394,106 1,423,150 1,452,194.
flow 4,648,036 .86 6.8 .78 4.6 .42 31 .19 .05 03
.85
Discount 1 0.93 0.87 0.82 0.77 0.73 0.67 0.62 0.58 0.54
factors
(I=7%)
Pv - 1,134,454 1,086,53 1,047,903 1,006,37 975,293.4 914,592 864,346 825,427 784,185

4,648,036 2 .24 0.4 5
.85

8. PLANT LOCATION AND PLANT LAY OUT

8.1. Site selection
Site selection is not an easy problem because if the selection is not proper then all money
spent on factory building, machinery and their installation will go as waste and the owner has
to suffer a great loss. Therefore, as plant designer we select the best site to erect our plant.
Our plant site location is at Debre berhan city, the specific site for this plant is tebase.
1. The selected site is near to raw material source, so that the transportation cost
to the site is low. As though the main raw material for the production of bio
ethanol is barley spent grain. The selected site is very comfortable to use this
resource.
2. The cost of transporting finished goods, advertising and distribution is reduced
since it is located near to the market.
3. Labor cost is an important item of the total production in manufacturing
and in the selected site labor is highly available and the cost of labor is very
cheap.
4. Electricity which is the main energy source of production is highly and nearly
available to the selected site.
5. Climatic and atmospheric conditions are governing factors to the successes of
production plants.
8.2. Plant lay out
Having selected a suitable site for the chemical plant, it is possible and necessary to make a
preliminary decision regarding the layout of the plant equipment. Although the equipment
has not been designed in detail, preliminary estimates of the physical size of each item should
be available in the equipment list. Any sizing differences between the initial and final
estimates should not be too excessive, and appropriate areas should be allowed around the
plant items when determining the layout.

Figure 8.1: plant lay out

9. ENVIRONMENTAL CONSIDERATION
This topic needs to be considered at an early stage in the design, in relation to the site
considerations, and reassessed during the detailed design stage. The effect of the operation of the
chemical plant upon the environment and the population must be considered during the
feasibility study and during subsequent design stages. The formal environmental impact analysis
(EIA) has two parts, these relate to:
The treatment of unwanted chemicals (by-products) and the concentrations of liquid
discharges and gaseous emissions during normal operation;
The handling of a major chemical accident, including all chemicals within the plant and
any subsequent reaction products, and containment and clean up. The environmental
impact analysis is specific to the plant under consideration; the comments/suggestions
made here are of a general nature.
9.1. General Considerations
All waste chemicals from the plant must be disposed of in an acceptable manner. Dumping of the
waste may not be allowed or, if it is, it may be prohibitively expensive. Some form of treatment,
e.g. dilution, neutralization, purification, separation, etc., may be necessary prior to disposal. It is
necessary to determine whether it is more economical (and preferable for the efficient operation
of the plant) to perform this treatment within the plant itself. Consideration should also be given
to the installation of separate drainage systems from certain sections of the plant and for
particular wastes, e.g. rainwater, domestic waste, relatively pure process water, acid spills.
Separate drainage systems are best installed during the plant construction stage.
This scheme will be more expensive than a central collection tank for all liquid waste, but it will
allow separate treatment and possible recycling of liquids. The concentrations of all chemicals to
be discharged, including gaseous emissions, must be determined and measures taken to ensure
that these levels conform to allowable legislative standards. The cost of clean-up can be high,
e.g. scrubbing systems, filters, etc., and may affect the economic analysis of the plant operation.
It is prudent to ensure that not only are present emission standards observed, but also that the
plant could conform to any subsequent legislative reductions in these emission levels (while still
operating economically).
The effects of all emissions on the environment and upon company employees and the
population must be assessed.

The second part of the environmental impact analysis (EIA) relates to the effects of a major
accident or spill within the plant. The safety aspects of an explosive gas discharge (for example)
should be considered in conjunction with the loss prevention studies for the plant. However,
proposals for containment, clean-up, and discharge of major chemical spills should be part of the
EIA report. Any proposals should ensure the safety of personnel, minimize the discharge and its
effect on the environment, and preserve the integrity of the plant. The worst situation should be
evaluated, not just the most likely scenario.
Factors to be considered include the quantity and location of chemicals stored within the plant,
prevailing wind direction, location of plant personnel and the general public, access to the plant
and to particular high-risk areas.

10. CONCLUSION AND RECOMMENDATION

10.1. Conclusion
This study shows that barley spent grain, which is by product of beer production, can be a good
source of ethanol (beverage and fuel) like that of molasses. From the experiment results it can be
seen, parameters such as yeast concentration and fermentation time have a significant effect on
alcohol content. When concentration of yeast increases, the yield of ethanol increase up to
100ml and then it starts to decrease also alcohol content was increases with the increase in
fermentation time and reaches maximum value for 3 day and it starts decreases. The Optimized
conditions for the experiment are: hydrolysis temperature and time 140oc for 25min, PH 4.5-5,
yeast concentration 100ml, and 3 day fermentation which gives maximum ethanol percentage of
48.3 %( v/v). Based on the rough economic analysis, production of bioethanol from Barley spent
grain is profitable since the rate of return on investment was 27.5% and the payback period is 3.7
years.
Implementation of this research work is environmental friendly and will get continuous feed
from brewery industries. As a result industries will expand in our country due to its push effect.

10.2. Recommendations
Further studies are recommended on
Further study of the Pretreatment, Fermentation and Distillation processes are

recommended to maximize the yield.
Alternative hydrolysis methods of ethanol such as enzymatic hydrolysis have to be done
in order to investigate the variation that could be arise on the quality and quantity of the
ethanol yield.
Further economic evaluation study in the production process is needed.
In our university some laboratory equipments were not fulfilled such as: purified
ethanol distillery, fermenter, flash point apparatus and gas chromatography which is
used to ethanol characterization; as result we do not achieved an expected purified or
good grade ethanol and not characterized simply. Therefore; we recommended that
the department should be satisfied all those equipment.
Barley spent grain used for different applications such as production of xylitol, furfural,
biogas and bioethanol so need further study.

References
"Abrief history of rocketry ." NASA historiccal archive, via science. nansa.gov.
Admas, K E, and T S Rans. "Adverse reactions toalcohol and alcoholic beverage ." Ann Allergy
Asthma immomunol, 2013: 439-445.
"Alcohol use and safe drinking ." US national institute of health .
Badwal, s.p s, S Giddey, A Goel Kulkarni, and Basu S J. "Direct ethanol fuel cell for transport
and stationary application ." Acompressible reviwe applied energy , 2015.
Braeuning, Robert A. "Rocket propellant." Rocket and space technology, 2006.
Campbell, C J, and J H Lahrrere. "The end of cheap oil." Sci Am, 1998: 78- 83.
"Can ethanol Fire places Be cozy." Wall street journal , 2016.
Cellulosic Ethanol. 2010.
Chandel, K A, E S Chan, Ravinder Rudravaram, L M Narasu, V L Rao, and Poqaku Ravindra.

"Economic and environmental impact of bioethanol production technologies." an appraisal,
Biotech and Molecular Biology Review, 2007: 14-32.
Demirbas, A. "Bioethanol from Cellulosic Materials." A Renewable Motor Fuel from , 2005:
327-337.
Gray, K A, L Zhao, and M Emptage. "Bioethanol." Curr. Opin. Chem. Biol, 2006: 141-146.
Harinen, S. "Analysis of The Top Phase Fraction of Wood Pyrolysis Liquids." Master’ Thesis,
Laboratory of Applied Chemistry, Department of Chemistry, University of Jyvaskyla, 2004.
Harris, J, A Baker, and J Zerbe. "Two stage dilute sulfuric acid hydrolysis of hardwood for
ethanol production." Energy Biomass Wastes, 1984: 1151-1170.
Hettenhaus, J. "Ethanol fermentation strains: present and future requirements for biomass to
ethanol commercialization." 1998.
Holtzapple, M T, and E Humphrey A. "The effect of Organosolv Pretreatment on the enzymatic

hydrolysis of Poplar." Bioethanol Bioeng, 1990: 670-676.

J, M Coulson, and F Richardson J. Chemical Engineering Vol 1,2,6 forth Edition press,
NewYork 2005.
Jones, R, N Pamment, and P Greenfield. "Alcohol fermentation by yeasts- the effect of

environmental and other variables." Proc. Biochem, 1981: 42-49.
Kunze, W. Technology Brewing and. Berlin: Mieth, H.O., 1996.
Licht, F O. "World Ethanol Market." The Outlook to 2015, Tunbridge Wells, AgraEurope
Special Report, UK , 2006.
Lin, Y, and S Tanaka. "Ethanol fermentation from biomass resources." current state and
prospects. Appl. Microbial, 2006: 627-642.
McDonnel, G RUssell AD. "Antiseptics and disinfectants." Microbial review, 1999.
McMillan, J. "Pretreatment of Lignocelluloses biomass. ." Conversion of hemicellulose

hydrolizates to ethanol, 1994: 292-324.
Methanol poisoning, National institute of health. 2015.
Nirbhay Gupta, Gupta. "Hydrolysis of Lignocellulosic Materials for Ethanol Production,

National Institute of Technology,Rourkela." 2008.
Onuki, S. Bioethanol, Industrial production process and recent studies. USA: Perry chemical
engineering hand book, 2005.
Parveen, Kumar. "Methods for Pretreatment of Lignocellulosic Biomass for Efficient Hydrolysis
and Biofuel Production." 2009.
Pongsawatmanit, R, T Temsiripong, and T Suwonsichon. "Thermal and rheologica lproperties of

tapioca starch and xyloglucan mixtures in the presence of sucrose." Food Res, 2007: 239-248.
Purwadi, R. "Continuous ethanol production from dilute-acid hydrolysates: ." Department of

Chemical and Biological , 2006.
Sanchez, G, L Pilcher, C Roslander, T modig, M Galbe, and G Linden. "Diluted acid hydrolysis
for Fermentation of the Bolivian straw material Paja Brava ." BioresourTechnol, 2004: 249-256.

Sofer, S, and O Zaborsky. "Biomass conversion process for Energy and Fuels." 637. Third
Eddition , Washington DC, 1981.
Solomon, B D. "Grain and cellulosic ethanol." History, economics, and energy policy. Biomass
& Bioenergy, 2007: 416-425.
Sun, Y, and J Cheng. "Hydrolysis of lignocellulosic materials for ethanol production:." Bioresour
Technol, 2002: 1-11.
Wheals, A E, L C Basso, D M.G Alves, and H V Amorim. "Fuel ethanol after 25 years."
TIBTECH, 1999: 482-487.
Zuccotti, Gain Vincenzo, and Valentia Fabiano. "Safety issue with ethanol as an exipient in
drugs intended for pediatric use." Expert opinion on drug safety , 2011: 449-502.

Appendices
Appendix A: Chemical used for yeast propagation
Appendix B: Prepared media for yeast growth
Appendix C: Sample After hydrolysis

Appendix D: Sample after fermentation
Appendix E: Final product

Bio Ethanolll Final PDF

Hochgeladen von

Dokumentinformationen

Copyright

Verfügbare Formate

Dieses Dokument teilen

Dokument teilen oder einbetten

Freigabeoptionen

Stufen Sie dieses Dokument als nützlich ein?

Sind diese Inhalte unangemessen?

Copyright:

Verfügbare Formate

Bio Ethanolll Final PDF

Hochgeladen von

Copyright:

Verfügbare Formate

DEBRE BERHAN UNIVERSITY DEPARTMENT OF CHEMICAL ENGINEERING

BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page i

List of figures ................................................................................................................................. vi

List of tables .................................................................................................................................. vii

List of Acronyms ......................................................................................................................... viii

1.1 Background of the study ....................................................................................................... 1

1.2. Statement of the problem ..................................................................................................... 2

1.3. Objectives of the study ......................................................................................................... 2

1.3.1. General objective ........................................................................................................... 2

1.3.2. Specific objective .......................................................................................................... 2

1.4. Significance of the study ...................................................................................................... 2

1.5. Scope of the project .............................................................................................................. 2

1.6. Limitation ............................................................................................................................. 3

2. LITERATURE REVIEW ........................................................................................................... 4

2.1. History of ethanol production .............................................................................................. 4

2.2. Bio-ethanol and its application ............................................................................................. 4

2.3. Ethanol and its characteristics .............................................................................................. 7

2.4. Feedstocks for bioethanol production .................................................................................. 8

2.4.1. Bioethanol from lignocelluloses feedstocks .................................................................. 9

2.5. Barley spent grain............................................................................................................... 11

2.5.1. Physicochemical properties of barley spent grain (BSG) ............................................ 11

2.5.2. Potential applications for barley spent grain ............................................................... 12

BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page ii

2.6. Bioethanol production process ........................................................................................... 13

2.6.1. Pretreatment ................................................................................................................. 13

2.6.2. Hydrolysis .................................................................................................................... 17

2.6.3. Fermentation ................................................................................................................ 19

2.6.4. Distillation ................................................................................................................... 20

2.6.5. Dehydration ................................................................................................................. 20

2.7. Technology selection.......................................................................................................... 22

2.8. Factors affecting hydrolysis of lignocellulosic materials ................................................... 23

2.8.1. Properties of the substrate............................................................................................ 23

2.8.2. The acidity of the system ............................................................................................. 23

2.8.3 Rate of decomposition of hydrolysis products during hydrolysis ................................ 24

2.9. Factors affecting fermentation ........................................................................................... 24

2.9.1. Effect of sugar concentration ....................................................................................... 24

2.9.2. Effect of temperature ................................................................................................... 24

2.9.3. Effect of pH ................................................................................................................. 25

2.9.4. Ethanol concentration .................................................................................................. 25

2.9.5. Osmotic tolerance ........................................................................................................ 25

2.9.6. Inhibitor tolerance........................................................................................................ 26

2.10. Previous study of bioethanol production .......................................................................... 26

3. MATERIAL AND METHODS ................................................................................................ 29

3.1. Materials and equipment used for the experiments ............................................................ 29

3.2. Experimental set up ............................................................................................................ 30

3.3. Experimental procedure ..................................................................................................... 31

3.3.1. Sample preparation ...................................................................................................... 31

3.3.2. Dilute acid pretreatment .............................................................................................. 31

BIOETHANOL PRODUCTION FROM BARLEY SPENT GRAIN Page iii

3.3.3. Filtration ...................................................................................................................... 32

3.3.6. Fermentation ................................................................................................................ 33

3.3.7. Ethanol Separation (Distillation) ................................................................................. 34

4. RESULT AND DISCUSSION ................................................................................................. 36

4.1. Moisture content determination ......................................................................................... 36

4.2. Ethanol characterization ..................................................................................................... 36

4.2.1. Specific gravity of ethanol ........................................................................................... 36

4.3. Effect of yeast concentration .............................................................................................. 37

4.4. Effect of fermentation time ................................................................................................ 39

5. MATERIAL AND ENERGY BALANCE ............................................................................... 41

5.1 Material balance .................................................................................................................. 41

5.2. Energy balance ................................................................................................................... 47

6. SIZING AND DESIGN OF MAJOR EQUIPMENTS ............................................................. 52

6.2. Design of process equipment ............................................................................................. 56

7. FINANCIAL EVALUATION .................................................................................................. 67