FPM 174

STUDY ON BIO-ETHANOL EXTRACTION PROCEDURE
FROM SWEET GOURD, SWEET POTATO AND MAIZE
MS Thesis
HUSSAIN MOHAMMAD KHALEDUJJAMAN

Roll NO: 12 FPM JJ07 M
Registration No.:34611
Session: 2007-2008
Semester: January-June, 2014
DEPARTMENT OF FARM POWER AND MACHIENARY

BANGLADESH AGRICULTURAL UNIVERSITY
MYMENSINGH-2202
JUNE 2014
A THESIS
BY

Roll NO: 12 FPM JJ07 M
Session: 2007-2008
Submitted to the
Department of Farm Power and Machinery
Faculty of Agricultural Engineering and Technology
Bangladesh Agricultural University, Mymensingh
In partial fulfillment of the requirement for the

Degree of
MASTER OF SCIENCE (MS)
IN
FARM POWER AND MACHINERY

MYMENSINGH-2202
JUNE 2014
A THESIS
BY

Roll NO: 12 FPM JJ 07 M
Session: 2007-2008
Approved as to style and content by
____________________________ _________________________
(Prof. Dr. Md. Daulat Hussain) (Dr. Muhammad Ashik-E-Rabbani)
Supervisor Co-supervisor
_______________________________
(Dr. Md. Rostom Ali)
Associate Professor & Chairman
Examination Committee and Head
Department of Farm Power and Machinery
Bangladesh Agricultural University
Mymensingh-2202

MYMENSINGH-2202
JUNE 2014
CONTENTS
Chapter No. Title Page No.
ACKNOWLEDGEMENT iv
ABSTRACT v
CONTENTS vi
LIST OF TABLES viii
LIST OF FIGURES ix
ABBREVIATION x
CHAPTER INTRODUCTION 1
1
1.1 Objectives 6
1.2 Specific Objectives 6
CHAPTER REVIEW OF LITERATURE 7
2
2.1 Raw Materials for Bio-ethanol Production 7
2.2 Bio- ethanol Production Methods 10
2.3 Bio- ethanol Extraction From Different Sources 15
2.4 Effect on Environment 16
CHAPTER CHARACTERISTICS OF MAIZE, SWEET GOURD AND 19
3 SWEET POTATO WITH ITS CHEMICAL ANALYSIS AND
TYPES OF ETHANOL EXTRACTION METHODS
3.1 Characteristics of maize 19
3.1.1 Distribution and habit 19
3.1.2 Botanical Feature 19
3.1.3 Uses 21
3.2 Characteristics of sweet gourd 23
3.3 Characteristics of sweet potato 27
3.4 Ethanol Extraction 28
3.4.1 Water Bath Type Evaporator 29
3.4.2 Rotary Evaporator 29
3.4.3 Oil Bath Type Evaporator 29
vi
CONTENTS (CONTINUED)
Chapter No. Title Page No.
3.4.4 Ethanol extraction 30

CHAPTER METHODOLOGY 31
4
4.1 Selection of sweet gourd, sweet potato and maize 31
4.2 Study area 31
4.3 Fermentation 33
4.3.1 Sweet gourd fermentation 34
4.3.2 Sweet potato fermentation 35
4.3.3 Maize fermentation 36
4.4 Total fermentation 37
4.5 Process of extraction 38
4.6 Collection of Bio-ethanol 39
CHAPTER RESULT AND DISCUSSION 40
5
5.1 Ethanol from Sweet gourd 40
5.2 Ethanol from sweet potato 40
5.3 Ethanol from maize 41
5.4 Comparison of percentages of different crops extracted Bio- 42
ethanol
5.5 Extraction process of Bio-ethanol 43
5.6 Chemical conversion process of Bio-ethanol 43
5.7 Chemical analysis of Bio-ethanol 44
5.8 Production cost of Extracted bio-ethanol 45
5.9 Price of Bio-ethanol 46
CHAPTER CONCLUSIONS AND RECOMMENDATIONS 47
6
6.1 Conclusions 47
6.2 Recommendations 48
CHAPTER REFFERENCES 49
7
vii
LIST OF TABLES
Table No. Title Page No.
1.1 Recent few years production of sweet gourd, sweet potato and maize 5
3.1 Gross chemical composition of different types of maize (%) 23
3.2 Gross chemical composition of Sweet gourd 26
3.3 Gross chemical composition of Sweet potato 28
4.1 Quantity of total fermentation 37
5.1 Sweet gourd fermentation 40
5.2 Ethanol Extraction in percentage 40
5.3 Sweet potato fermentation 41
5.5 Ethanol from maize 41
5.7 According to market price of those crops and its Bio-ethanol percentage 42
5.8 Chemical analysis 44
5.9 Fixed cost for conducting experiment (For 3 years) 45
5.10 Price of Bio-ethanol 46
viii
LIST OF FIGURES
Figure No. Title Page No.
4.1 Study Area 32

4.2 Sweet gourd fermentation 35
4.3 Sweet potato fermentation 36
4.4 Maize fermentation 37
4.5 Quantity of total fermentation 38
4.6 Before extraction 38
4.7 Evaporation system by using heater 39
4.8 Collection of Bio-ethanol 39
5.1 Comparison of percentages and prices of different crops extracted Bio-ethanol 42
5.2 Bio-Ethanol extraction process 43
5.3 Starch to Ethanol process 44
ix
ABSTRACT
This experiment was conducted to bring in a suitable and economic bio-ethanol

extraction method for sweet gourd, sweet potato and maize growers in
Bangladesh. It was carried out during the periods from January to April 2014 in
the department of Farm Power and Machinery, BAU and Saidpur upazila of
Nilphamari District. During the experiment three types of samples such as sweet
gourd, sweet potato and maize were selected for their availability and low cost in
this zone (Nilphamari). Three steps were followed for Bio-ethanol extraction.
These were preparation of raw materials, fermentation and ethanol extraction.
Mainly an ethanol extraction procedure was maintained for this experiment which
can control the temperature at 79c for extracting the raw ethanol from fermented
solution. From this extraction 19%, 21% and 22% Bio-ethanol was collected from
sweet gourd, sweet potato and maize respectively. The highest concentration of
ethanol has been observed in maize about 60%, others were 46% and 32% ethanol
from sweet gourd and sweet potato respectively in this experiment. Using this
extraction method, 95 ml, 105 ml, 110 ml of bio-ethanol was extracted from 500
ml sweet gourd, sweet potato and maize each. It was found from the calculation
that the developed system can bring yearly benefit of Tk. 32000/ton,
Tk.33666/ton, Tk. 35000/ton with benefit-cost ratio 1.193, 1.207, 1.197
respectively for the three samples. This development, therefore, has proven very
beneficial for Bangladeshi bio-ethanol extractors.
v
ACKNOWLEDGEMENT
All praises are due to the Almighty ALLAH who is the creator of all things seen or
unseen. The author also makes sukria to ALLAH for enabling him to complete this
work successfully.
The author expresses profound appreciation, heartfelt gratitude and immense

indebtedness to his respectable teacher and supervisor Dr. Md. Daulat Hussain
professor, Department of Farm Power & Machinery, Bangladesh Agricultural
University (BAU), Mymensingh for his scholastic guidance, supervision, instruction,
constructive criticism and constant encouragement throughout the whole period of this
work
The author sincerely expresses his profound gratitude to his reverend co-supervisor Dr.
Muhammad Ashik-e-Rabbani, Associate professor, Department of Farm Power &
Machinery BAU, Mymensingh for his guidance, kind and precious suggestions and
immense help during the entire period of the work.
The author is also grateful to Dr. Md. Rostom Ali, Associate professor and Head,
Department of Farm Power & Machinery, BAU, Mymensingh for his cooperation,
encouragement and providing departmental facilities for this study.
The author likes to extend his heartfelt appreciation and thanks to all other respected
teachers of the Department of Farm Power & Machinery, BAU, Mymensingh for their
encouragement and valuable suggestions to complete this work.
The author is grateful to his beloved parents, friends Nasim, Joy,Rubel, Habib, Josim,
Sudab, Emon and all family members for their encouragement and blessings. Also
thanks to Humayun kabir, Student, Chemistry department. Rajsahi University for lab
facilities of RU.
The author
iv
CHAPTER I
INTRODUCTION
Ethanol is a desirable fuel additive because it allows fuel to burn more cleanly and
lowers greenhouse gas emissions. It is cost-effective to blend ethanol into gasoline
in view of high crude oil prices in recent years (Louime and Uckelmann, 2008).
The idea to use ethanol as a source of energy is not new. The oldest evidence
about alcohol used as an engine fuel comes from 1899. Between the world wars
about 4 million cars used gasoline blended with 25% volume of ethanol
(Chereminisoff, 1979). Several methods have been identified to produce
bioethanol. Bioethanol can be synthesized from cellulose and hemicellulose that
originates from the many sources of biomass (Cheng et al., 2007). Bioethanol is
one form of renewable energy source that is fast gaining foot hold as potential fuel
to power automotive engine. Microscopic yeast cells break down the starch and
water, creating the so called Bioethanol and carbondioxide as end products.
Bioethanol burns to produce carbondioxide and water in complete combustion, a
process akin to gasoline (Mohammad, 1999). In an earlier study (Taherzadeh,
1999), physiological effects of inhibitors on ethanol from lignocellulosic materials
and fermentation strategies was comprehensively investigated. Yeast based
fermentation, for example, has yielded ethanol from sugar or crops. The
production of ethanol by fermentation of sugar has already been commercially
established, but innovative studies could bring improvements to reactors and
separation systems. To produce ethanol from lignocellosic materials, it is essential
to hydrolyze it before fermentation at the demonstration and industrial level.
Enzymatic hydrolysis is still at an early stage, requiring substantial fundamental
research (e.g., for increased yields) (Kligtik and Demirbas, 1997). .
Bioethanol is an environmentally friendly fuel for vehicles that normally run on
petrol. As a renewable source of energy, it reduces demand on fossil fuels while it
burns more cleanly and with reduced emissions of CO 2, a greenhouse gas. As an
energy source, bioethanol is carbon neutral in that it reduces, by up to 70 %, the
1
amount of greenhouse gas released into the atmosphere. The CO 2 released during
ethanol production and combustion in an engine has already been absorbed from
the atmosphere during the growth of the crops due to photosynthesis.
Currently there is a big push to find and develop alternative sources of energy so
that dwindling reserves of crude oil and other fossil fuels may be conserved. As
Edward Teller, one of the Americas leading physicists points out: "No single
prescription exists for a solution to the energy problem. Energy conservation is
not enough. Petroleum is not enough. Coal is not enough. Nuclear energy is not
enough. Solar and geothermal energy are not enough. New ideas and
developments will not be enough by themselves. Only the proper combination of
all of these will suffice", it showed the importance of alcohol extraction as a fuel
for engine. Sweet potatoes are cooked and converted in a manner similar to
potatoes with the exception that they contain only about 66% water and some
dilution is necessary. Sweet potato contains saccharine (sugar) materials in which
the carbohydrate (the actual substance from which the alcohol is made) is present
in the form of simple, directly fermentable six and twelve carbon sugar molecules
such as glucose, fructose, and maltose. Hence, sweet potato has the potential for
alcohol production.
Humans have been producing ethanol for thousands years. According to the time
flow, the area of ethanol has been extending dramatically. The very first time,
ethanol existed only in alcoholic drinks. After some purification methods were
established, the usage of ethanol highly extended. Then, now, ethanol is at the
new step. The current increase in the gas price and interest in environmental
problems, ethanol becomes highly attractive again. However, to use ethanol as a
substitute of gasoline, there are so many problems on it. We have to establish
large industrial ethanol plant. Also, an increase in the productive efficiency will
be required
As demand for the limited global supply of nonrenewable energy resources

increases, the price of oil and natural gas keep increasing. A new biotechnological
2
approach for the production of ethanol by fermentation from the renewable
carbohydrate materials for use as an alternative liquid fuel has been attracting
worldwide interest (Ward and Singh 2002). Thus, there is a growing interest to
find alternative bioresearches other than sugarcane/beet molassess and starchy
crops such as cassava, sweet potato, and sweet sourghum for ethanol production.
Bioethanol is a microbiological way of converting simple sugar into ethanol and

carbon dioxide (CO2) ( Damaso, Castro and Adrade, 2004). Bioethanol is a
principal fuel that can be used as petrol substitute for vehicle (Aro etal., 2005). It
is a renewable energy source produced mainly by sugar fermentation process,
although it can also be manufactured by the chemical process of reacting ethylene
with steam (Anuj etal., 2007). The main sources of sugar required to produce
ethanol come from fuel or energy crops (Kim etal., 2005). These crops include
maize, cassava and cassava products, wheat crops, waste straw, guinea corn husk,
rice husk, millet husk, sawdust and sorghum plant. Ethanol is a high octane fuel
and has replaced lead as an octane enhancer in petrol (Oghgren etal., 2006). By
blending ethanol with gasoline we can also oxygenate the fuel mixture so it burns
more completely and reduces pollution emission. Ethanol fuel trends are widely
sold in the United State. The most common blend is 10% ethanol and 90% petrol
(E10) and vehicle engines require no modification to run on E 10 and vehicle
warranties are unaffected also (Lin, T., 2006.). Ethanol derived from biomass is
the only liquid transportation fuels that do not contribute to the greenhouse gas
effect (Nicholas et.al., 2002, Kroumov et.al., 2006, and Anuj and et.al., 2007).
Ethanol has been produced in batch fermentation with fungi strains such as
Aspergillus niger, Mucor mucedo, Saccharomyces cerevisiae that cannot tolerate
high concentration of ethanol (Ledward et.al., 2003, Oyeleke et.al., 2008 and
Seema et.al., 2007).
3
Recent few years in Bangladesh the production of sweet gourd, sweet potato and
maize are increasing rapidly. Most of the crops are used in poultry sector. But its
economical value is less. So here is an opportunity to move the production in Bio-
ethanol sector. The comparison of production of those crops in recent few years
are shown in Table 1.1
4
Table 1.1 Recent few years production of sweet gourd, sweet potato and maize in Bangladesh:
Sl. Crop
2009 2010 2011 2012 2013
no name
Area
Production Area Production Area Production Area Production Area Production
(1 lakh (1 lakh (1 lakh (1 lakh (1 lakh (1 lakh (1 lakh (1 lakh (1 lakh

(1 lakh M.ton) ha) M.ton) ha) M.ton) ha) M.ton) ha) M.ton)
ha)
1 Maize 1.74 11.37 2.02 13.70 2.40 16.42 2.92 20.42 3.12 21.78
Sweet
2 0.5 6.60 0.53 7.22 0.50 6.88 0.45 6.20 0.46 7.64
potato
Sweet
3 .053 1.82 .049 1.74 .061 2.20 0.72 2.55 0.69 2.49
gourd
Source: Agriculture Diary 2014 and report of DAE
This table elaborate that the amount of production of this three crops is gradually increasing day by day. So here is an
opportunity to develop the process for big aspect.
5
Some countries specially USA, UK, Brazil, Philippine, Korea, Japan, Africa etc extract
ethanol from different fruits, vegetables and root crops. Those countries already
producing bio-fuel in different way by extract ethanol from fruits, vegetables and root
crops. Bangladesh recently awareness has grown among the researchers and policy
planner to investigate into the feasibility of ethanol extraction from different agricultural
product. There are many other traditional or nontraditional fruits, vegetables and root
crops are available in Bangladesh and these should be research for their potentiality to
extract ethanol which use as bio-fuel. Based on the above study, the study was carried out
with the following objectives:
1.1 Objectives:
The main objective of the study is to develop technique to extract Bio-ethanol
from sweet gourd, sweet potato and maize by evaporating method
1.2 Specific objectives are:

1. To develop a technique to extract the Bio-ethanol from available sweet
gourd, sweet potato and maize.
2. Chemical analysis of sweet gourd, sweet potato and maize and also find out
the percentages of Bio-ethanol by experiment.
6
CHAPTER II
REVIEW OF LITERATURE
The purpose of this chapter is to discuss the available literatures related to ethanol
extraction from traditional fruits, vegetables and root crops which use as bio-fuel.
Very limited works have been done in Bangladesh in this regard. Works relevant
to the ethanol extraction performed by various researchers are reviewed in this
section.
2.1 Raw Materials for Bio-ethanol Production
In Brazil Energy Business Daily; (2007) reported that ethanol fuel is produced
from sugar cane which is a more efficient source of fermentable carbohydrates
than corn as well as much easier to grow and process. Beside that they also
produced ethanol from different fruit, vegetable and root crops.
Eduardo de Sousa, and Marco Patina, from Brazil (2007) , reported that each
country should not only determine its capacity to produce raw materials for bio-
fuels, but should also continue using the land reserved for agriculture to produce
food. In most cases, including Brazil, they claimed, there is enough land for
agriculture to produce raw materials for bio-fuels without endangering food
security. Another important aspect is the fact that industry is the driving force to
reduce hunger. As jobs are being created in service sectors to support industry, this
will help people out of poverty, studied Sousa. He added that small farmers may
sell their cassava and generate financial resources to buy other foodstuffs and still
have enough of it for their normal diet.
Maputo Mozambican (2007) scientists and researchers told an International

Symposium on Tropical Roots and Tubers that they are determined to develop
7
varieties of cassava appropriate for the production of ethanol. They were speaking
in Maputo on the theme Roots and Tubers for the production of ethanol,
Challenges and Opportunities.
The national coordinator of the Roots and Tubers Program, Fernando Chitio,
(2007) reported that research is being prepared to identify varieties of cassava for
the production of ethanol. They are interested in using cassava for the production
of ethanol, because that will add value to cassava itself. With annual production of
about six million tones, Mozambique is the sixth largest producer of cassava in
the world. Despite the apparent abundance of cassava, and of land on which it can
be grown, there are voices saying that the production of ethanol will endanger
Mozambican food security and biodiversity. The fear is that farmers in countries
with tropical climates may dedicate their full attention to the cultivation of plants
used for the production of ethanol to the detriment of their own food security.
Chitio dismissed such fears, arguing that farmers will not cut their own throats,
but will only sell surplus cassava for ethanol, after taking what they need for their
own consumption. As raw material for industry, cassava would attract investment
and stimulate productivity and it also offers the farmers an opportunity to sell it as
a cash crop.
ISU reported that (2006) researchers of Iowa State University have used mold to
convert corn fiber into ethanol. The discovery could turn byproducts of corn
milling into another source of fuel.
PESN studied that (2006) Iogen's cellulose ethanol is a fully renewable, advanced
bio-fuel made from the agriculture product such as cereal straws, barely, sweet
gourd, jack fruit, pineapple and corn stover and is one of the most cost-effective
ways to reduce greenhouse gas emissions in road transport.
8
In Nigeria, the Federal Government of Nigeria, the Niger Delta Development
Commission (NDDC), Shell Petroleum Development Company (SPDC) of
Nigeria, United States Agency for International Development (USAID), and
States in southern Nigeria. (2005) IIT reported that local production of ethanol
from maize, guinea corn, millet, other starchy substrates, and cellulose is as old as
the country itself. Apart from food and pharmaceutical uses, ethanol is finding
itself alternative uses for bio-fuel in most of the developed world.
Reuters studied (2005) ethanol export contracts from Brazil are looking more like
contracts for other goods, such as sugar, soybean and oil a sign that the renewable
fuel may be on its way to becoming a world commodity.
News wise reported that (2005) researcher of University of Florida has developed
a biotechnology bug that converts fruit, vegetable and root crops wastes into
ethanol.
Renewable Energy Access developed (2005) a biomass Plant to Convert rice
straw to ethanol. Colusa Biomass Energy, a biomass-to-ethanol company, has
engaged technical assistance consultants for a production plant in the Sacramento
Valley that will convert rice straw to chemical products and ethanol.
Vegetable by-products have mainly been utilized as a source of dietary fiber and
natural antioxidants, but residual vegetables could also be used as a raw material
for soluble sugar and polyol extraction. Therefore, 2 celery (Apium graveolens)
residues from the food industry composed either of stalks alone or stalks plus
leaves, were extracted with hot 85% ethanol to solubilize soluble sugars and
mannitol. Low-molecular-weight carbohydrates in the extracts were identified and
quantified by high-performance liquid chromatography. Both celery residues
contained similar amounts of sucrose (5.75.9%), but different ratios of hexoses
(glucose and fructose) to mannitol. Total sugar content and mannitol were higher
9
in the stalks (45.5% and 15.2%, respectively) than in the stalk plus leaf residues
(33.9% and 13.3%, respectively). Mannitol represented 33.539.3% of the total
carbohydrate in celery wastes. The alcoholic extracts from celery residues are
proposed as a natural source of mannitol and soluble sugars, which might be used
by the food industry. The alcohol-insoluble residue from celery by-products could
be further used for the preparation of dietary fiber-rich food supplements and also
use as bio-fuel (Ruperez and Toledano, 2003).
2.2 Bio- ethanol Production Methods
Islam (2009) conducted an experiment on mechanical extraction method (

Scraping for leaf fibre) for fermentation and extracted fibre from Dhaincha
(Sesbania aculeata), Ladys finger (Abelmoschus esculentus), Banana pseudo
stem (Musa indica), Pineapple leaf (Ananas comosus) and Sisal leaf (Agave
sisalana). Four types of treatments were used for findings suitable fermentation
technique, i) aerobic fermentation ii) anaerobic fermentation iii) fermentation with
urea iv) fermentation with CaO. With the increase of fermentation time, diameter
of fibre decreased in all the treatments and as a result the tensile strength also
decreased. The estimated fibre production of Dhaincha was the highest 1900
kg/ha and then sisal leaf, pineapple leaf, ladys finger and banana pseudo stem,
was 1817 kg/ha, 1459 kg/ha, 220 kg/ha and 197kg/ha, respectively. Study
revealed that Dhaincha fibre will be better among the observed samples.
Avril Rodiel Bries (2008), submitted an investigatory project report in 16 TH

international environmental project Olympiad (16 TH INEPO), the project was
conducted to extract bio-ethanol from pineapple peelings through the
Simultaneous Saccharification and Fermentation (SSF) process using the yeast
Saccharomyces cerevisiae . On average, 435,000 metric tons of pineapples are
produced annually in the Philippines, which is one of the countrys leading
10
commercial fruit products. However, there are a lot of unused excess parts of the
pineapple, notably the peelings, which are considered as waste and contribute to
the countrys garbage problem. One solution to both dilemmas is to produce bio-
ethanol from pineapple peelings. This will be made possible by extracting the
glucose content of the peelings and fermenting it through the process of
Simultaneous Saccharification and Fermentation (SSF) using the yeast sample.
Peel samples, coming from pineapple, will undergo a series of physical and
chemical analyses to determine the glucose content, which will be utilized to yield
bio-ethanol. The SSF process will be manipulated in terms of fermentation time
and buffer level used to determine the best variables for ethanol production. This
project entitled the extraction of bio-ethanol from Pineapple peelings through
Simultaneous Saccharification and Fermentation using the Yeast Saccharification
cerevisia will indeed be a great help to the economy and the environment.
Owing to the high starch content (55-65% on dry weight basis) and organic matter
of cassava wastes, an attempt has been made to utilize it for the production of
three bio-products, i. e. alpha -amylase, lactic acid and ethanol in solid substrate
fermentation by incubating the solid residue at different moisture holding capacity
(40-80%) and incubation period (12-60 hr for alpha -amylase, 24-144 hr for
ethanol and 2-10 days for lactic acid). The highest product yield was obtained at
60% moisture holding capacity of the residue and period of incubation varied
from 36 hr ( alpha -amylase), 120 hr (ethanol) to 6 days (lactic acid). This study
showed that the solid residues from cassava starch factories could serve as a low-
cost substrate for bio -products production (Ray et al., 2008).
Eco Geek studied (2007) (Slashdot discussion) the cellulosic ethanol process
produces ethanol from cellulose, which all plants have, instead of from sugar,
which is only abundant in corn, fruit ,vegetables and root crops. Corn ethanol only
produces 1.3 units of energy for every unit of energy that goes into growing the
11
crop and converting the sugar to ethanol. Cellulosic ethanol can produce as much
as 16 units of energy for every one unit of energy put into the process.
Renewable Energy Access reported that (2007) new technologies are required to
produce ethanol from biomass cellulose rather than from the fermentation of
valuable grains. Lignol Energy Corporation has acquired and modified a solvent-
based pre-treatment technology originally developed by a subsidiary of General
Electric (GE).
Sattar et al. (2007) investigated the some decorative plants widely grown in
Bangladesh. The plants Mimusops elengi, Madhuca indica, Hiptage benghalensis
and Polyalthia longifolia have been analyzed for their furfural contents by
colorimetric spectrophotometry after hydrolysis in 13 percent HCl and extraction
with 50% ethanol. The results revealed that the furfural concentration in these
plants is in the range of 3.9-10.3 percent, indicating the potential of these plants as
good source of pentosans and furfural. TLC and conventional chemical test
further confirmed the presence of furfural in extracted solution.
Impact Lab reported that (2006) a new method developed by Honda and the
Research Institute of Innovative Technology for the Earth (RITE) allows large
volumes of cellulosic ethanol to be produced from widely available waste wood,
rice straw, leaves and some of fruits vegetables such as cassava, pineapple
jackfruit etc . A microorganism developed by RITE helps reduce interference in
the fermentation process, allowing for far more efficient ethanol production.
Kabir (2006) carried out an experiment to find out the prospects of biodiesel
production from Jatropha curcas. During the experiment plant growth, number of
flowers per plant, number of fruits per plant, number of seeds per plant, physical
characteristics of Jatropha fruit and seed were studied. Oil extraction from
12
Jatropha seed through chemical method was found to be higher than the
mechanical method. But it was more expensive than the mechanical method. The
percentage of oil content in Jatropha seed was found to be almost 36% (w/w) and
the estimated price of one liter biodisel from Jatropha seed was nearly Tk. 34.50.
Terrell et al. studied (2005) the ethanol is being used in reformulated gasoline
(RFG) and oxygenated gasoline to fulfill the requirements of the Clean Air Act.
Currently, there are 56 ethanol production facilities in the United States,
producing approximately 4 billion gallons of ethanol per year, with more under
construction and expansion. Most of the ethanol is produced from the starches
found in corn. Because ethanol poses no serious health risk to humans, it is the
most likely replacement of methyl tertiary butyl ether (MTBE), the most common
additive used in reformulated gasoline (RFG). MTBE has been ban in several
states and in other states; its concentration is limited due to health risks related to
MTBE found in drinking water. These starches are converted to sugar and the
sugar is fermented into ethanol. Ethanol fermentation is an inhibited-product
reaction, and inhibition limits productivity. Their focus is not necessarily the
production of ethanol, but rather the productivity. If ethanol were extracted from
the process as it is produced, it would reduce inhibition and increase productivity.
This project involves two methods that are aimed at improving the productivity.
One method uses a packed bed distillation column to absorb water and the other
method uses a membrane contactor to extract the ethanol. The two methods will
be analyzed and compared. The concepts, experiments, and ideas behind the two
methods are discussed in subsequent sections.
Teng-WenYuh et al. ( 2005) reported the efficiency of carbon dioxide

supercritical fluid extraction (SFE) of the biologically active compounds
imperatorin, meranzin and meranzin hydrate from the fruit peel of Citrus maxima
Merr. has been compared with that of solvent extraction with acetone. Under the
best SFE conditions tested for the three coumarone, which involved extraction at
13
50 degrees C and 27.6 MPa, the extractive efficiencies were 84, 76 and 18% for
imperatorin, meranzin and meranzin hydrate, respectively. The presence of
modifiers significantly affected the extraction efficiency: the highest extraction
efficiency of the three coumarone was obtained with ethanol as modifier.
Lautenschlaeger. Friendrich k (2004) studied the treatment of fruit, vegetable ,

root crops and meat product to extract ingredients there from. Some of the
chemicals which may be extracted by the different method of the present
invention are higher alcohols, particularly those having three or more carbon
atoms, such as fusel oil (i.e. butyl alcohol, iso - amyl alcohol and higher
homologues) and phenyl ethanol, esters such as ethyl acetate, methyl anthranilate,
iso amyl acetate, ethyl caproate, ethyl caprylate, ethyl caprate and phenyl ethyl
acetate, organic acids, especially volatile organic acids such as acetic acid and
sorbic acid, sulphidic compounds such as free sulphur, sulphur dioxide and
complex sulphides, and biogenic amines such as putrescence. Stability against
mould-growth may be improved by removal or neutralization of mould spores,.
Pesticide residues may be extracted from fruit and vegetable juices, and fusel oil
may be extracted from fermented and distilled alcoholic beverages.
In 2003 Ruperz et.al., studied the vegetable by-products have mainly been
utilized as a source of dietary fiber and natural antioxidants, but residual
vegetables could also be used as a raw material for soluble sugar and polyol
extraction. Therefore, 2 celery (Apium graveolens) residues from the food
industry composed either of stalks alone or stalks plus leaves, were extracted with
hot 85% ethanol to solubilize soluble sugars and mannitol. Low-molecular-weight
carbohydrates in the extracts were identified and quantified by high-performance
liquid chromatography. Both celery residues contained similar amounts of sucrose
(5.75.9%), but different ratios of hexoses (glucose and fructose) to mannitol.
Total sugar content and mannitol were higher in the stalks (45.5% and 15.2%,
respectively) than in the stalk plus leaf residues (33.9% and 13.3%, respectively).
14
Mannitol represented 33.539.3% of the total carbohydrate in celery wastes. The
alcoholic extracts from celery residues are proposed as a natural source of
mannitol and soluble sugars, which might be used by the food industry. The
alcohol-insoluble residue from celery by-products could be further used for the
preparation of dietary fiber-rich food supplements and also use as bio-fuel.
2.3 Bio- ethanol Extraction From Different Sources
Among the fermentation condition, SSF is found to more advanced and effective
technology for the microbial production ethanol, using different substrates such as
mahua flower (Mohanty et al. 2009), sweet sorghum (Kargi et al. 1985), apple
pomace (Ngadi and Correia 1992), rice straw (Roslan et al. 2011), sugarcane
bagasse (Shaibani et al. 2011) by S. cerevisiae.
Sweet potato is a tropical and temperate regions crop, normally found in Indian
sub-continent. It is used as a vegetable in the state of Odisha (Attaluri et al. 2010).
Ethanol fuel is produced from sugar cane which is a more efficient source of
fermentable carbohydrates than corn as well as much easier to grow and process
.Beside that they also produced ethanol from different fruit, vegetable and root
crops.( Brazil Energy Business Daily, 2007).
Traditionally, the yeast, Saccharomyces cerevisiae has been used all over the
world as the major ethanol producing microorganism (Lin and Shuzo 2006).
Ethanol export contracts from Brazil are looking more like contracts for other
goods, such as sugar, soybean and oil a sign that the renewable fuel may be on its
way to becoming a world commodity (Reuters, 2005).
15
Cassava and sago starch were evaluated for their feasibilities as substrates for
ethanol production using Zymomonas mobilis, ZM4 strain. Before fermentation,
the starch materials were pretreated employing two commercial enzymes,
Termamy l (thermostable -amylase) and AMG (amyloglucosidase). Using 2 l/g
of Termamyl and 4 l/g of AMG, effective conversion of both cassava and sago
starch into glucose was found with substrate concentration up to 30%(w/v) dry
substances. Fermentation study performed using these starch hydrolysates as
substrates resulted in ethanol yield at an average of 0.48g/g by Z. Mobilis, ZM4
(Rhee et al. 2004).
In recent years, however, research is focused on processes involving amylolytic

mold Trichoderma sp. As coculture with S. cerevisiae, because of several better
fermentation attributes as conversion of complex form of carbohydrates in to
glucose and then conversion of glucose to ethanol and CO 2 (Azevedo et al. 2000).
Sweet potato (Ipomoea batatas L.) represents an important biomass resource for
fuel alcohol production, because of its chemical composition and high density of
starch, compared to other forms of biomass, and thus premise as an alternative
bioresource for the production of ethanol through fermentation (Hang, et al. 1981,
1986; Roukas 1994).
Sweet potato is cheap, readily available in the local market and offers ease in
product processing. It contains starch (178 g /kg), total sugars (26 g /kg) and
protein (3.2 g/kg) on fresh weight basis (Tian et al. 1991).
2.4 Effect on Environment

The agricultural biomass includes branches, bushes, canes, corn and corn husks,
energy crops, forests, fruits, root crops, flowers, grains, grasses, herbaceous crops,
leaves, bark, needles, logs, roots, saplings, short rotation woody crops, shrubs,
switch grasses, trees, vegetables, vines and hard and soft woods (not including
woods with deleterious materials).It also includes organic waste materials
16
generated from agricultural processes including farming and forestry activities,
specifically including forestry wood waste. Agricultural biomass may be any of
the aforestated singularly or in any combination or mixture thereof. Biomass
includes materials containing cellulose, hemicellulose, lignin, protein and
carbohydrates such as starch and sugar. Common forms of biomass include trees,
shrubs and grasses, corn and corn husks as well as municipal solid waste, waste
paper and yard waste. Biomass high in starch, sugar or protein such as corn,
grains, fruits and vegetables are usually consumed as food. Conversely, biomass
high in cellulose, hemicellulose and lignin are not readily digestible and are
primarily utilized for wood and paper products, fuel, or are disposed of. Ethanol
and other chemical fermentation products typically have been produced from
sugars derived from feedstocks high in starches and sugars. While the discussion
of fermentation products describes the method and apparatus of the current
invention using the example of how the current invention is used to produce
ethanol, it will be appreciated by those skilled in the art that the current invention
also has application to other fermentation products, including citric acid, butanol
and isopropanol and that references to ethanol encompass these other fermentation
products. Recently, the conversion to ethanol of polymeric hexose and pentose
sugars in cellulose and hemicellulose has been achieved. It is an object of the
present invention to provide a method and apparatus for the production of ethanol
and other fermentation products from biomass containing deleterious materials
therein. It is another object of the invention to provide a method and apparatus for
the production of ethanol and other fermentation products from biomass
containing deleterious materials by effectively removing these constituents. It is
another object of the invention to provide a method and apparatus to blend
biomass containing deleterious materials with biomass substantially free of these
constituents, thereby producing blended biomass in which deleterious materials
are effectively diluted such that conversion to ethanol and other fermentation
products can be realized. It is another object of the invention to provide a system
17
for the separation and classification of biomass material for purposes of recycling
at least a portion of the material while preparing the remainder for conversion to
ethanol and other fermentation products or alternative disposal. These and other
objects of the invention are provided by a method and apparatus in which biomass
or non-virgin biomass is converted to ethanol and other fermentation products.
The non-virgin biomass initially contains deleterious materials which prevent or
inhibit its conversion to ethanol and other fermentation products (Van Draanen et
al., 1999)
18
CHAPTER III
CHARACTERISTICS OF MAIZE, SWEET GOURD AND SWEET

POTATO WITH ITS CHEMICAL ANALYSIS AND TYPES OF
ETHANOL EXTRACTION METHODS
3.1 Characteristics of maize
3.1.1 Distribution and habit
Maize, known in some English-speaking countries as corn, is a large grain plant

domesticated by indigenous peoples in Mesoamerica in prehistoric times. The
leafy stalk produces ears which contain the grain, which are seeds called kernels.
Zea mays, is an annual grass in the family Poaceae and is a staple food crop
grown all over the world. Maize is widely cultivated throughout the world, and a
greater weight of maize is produced each year than any other grain. The United
States produces 40% of the world's harvest; other top producing countries include
China, Brazil, Mexico, Indonesia, India, France and Argentina. Worldwide
production was 817 million tons in 2009more than rice (678 million tons) or
wheat (682 million tons). In 2009, over 159 million hectares (390 million acres)
of maize were planted worldwide, with a yield of over 5 tones/hectare. Production
can be significantly higher in certain regions of the world; 2009 forecasts for
production in Iowa were 11614 kg/ha. There is conflicting evidence to support the
hypothesis that maize yield potential has increased over the past few decades.
This suggests that changes in yield potential are associated with leaf angle,
lodging resistance, tolerance of high plant density, disease/pest tolerance, and
other agronomic traits rather than increase of yield potential per individual plant.
3.1.2 Botanical Feature

Scientific classification
Kingdom: Plantae
Order: Poales
Family: Poaceae
19
Subfamily: Panicoideae
Genus: Zea
Species: Z. mays
The maize plant is often 2.5 m (meters) (8 ft) in height, though some natural
strains can grow 12 m (40 ft). The stem has the appearance of a bamboo cane and
is commonly composed of 20 internodes of 18 cm (7 in) length. A leaf grows
from each node, which is generally 9 cm (3.5 in) in width and 120 cm (4 ft) in
length.
Ears develop above a few of the leaves in the midsection of the plant, between the
stem and leaf sheath, elongating by 3 mm/day, to a length of 18 cm (7 in)
(60 cm/24 in being the maximum observed in the subspecies.They are female
inflorescences, tightly enveloped by several layers of ear leaves commonly called
husks. Certain varieties of maize have been bred to produce many additional
developed ears. These are the source of the "baby corn" used as a vegetable in
Asian cuisine.
The apex of the stem ends in the tassel, an inflorescence of male flowers. When
the tassel is mature and conditions are suitably warm and dry, anthers on the tassel
dehisce and release pollen. Maize pollen is anemophilous (dispersed by wind),
and because of its large settling velocity, most pollen falls within a few meters of
the tassel.
Elongated stigmas, called silks, emerge from the whorl of husk leaves at the end
of the ear. They are often pale yellow and 7 in (178 mm) in length, like tufts of
hair in appearance. At the end of each is a carpel, which may develop into a
"kernel" if fertilized by a pollen grain. The percarp of the fruit is fused with the
seed coat referred to as "caryopsis", typical of the grasses, and the entire kernel is
often referred to as the "seed". The cob is close to a multiple fruit in structure,
except that the individual fruits (the kernels) never fuse into a single mass. The
20
grains are about the size of peas, and adhere in regular rows around a white, pithy
substance, which forms the ear (maximum size of kernel in subspecies is
reputedly 2.5 cm/1 in. An ear commonly holds 600 kernels. They are of various
colors: blackish, bluish-gray, purple, green, red, white and yellow. When ground
into flour, maize yields more flour with much less bran than wheat does. It lacks
the protein gluten of wheat and, therefore, makes baked goods with poor rising
capability. A genetic variant that accumulates more sugar and less starch in the ear
is consumed as a vegetable and is called sweet corn. Young ears can be consumed
raw, with the cob and silk, but as the plant matures (usually during the summer
months), the cob becomes tougher and the silk dries to inedibility. By the end of
the growing season, the kernels dry out and become difficult to chew without
cooking them tender first in boiling water.
Corn tassel Corn silks Corn ear Ripened corn
3.1.3 Uses
Human food
Maize is a major source of starch. Cornstarch (maize flour) is a major ingredient
in home cooking and in many industrialized food products. Maize is also a major
source of cooking oil (corn oil) and of maize gluten. Maize starch can be
hydrolyzed and enzymatically treated to produce syrups, particularly high-
fructose corn syrup, a sweetener; and also fermented and distilled to produce grain
alcohol. Grain alcohol from maize is traditionally the source of Bourbon whiskey.
Maize is sometimes used as the starch source for beer. Within the United States,
the usage of maize for human consumption constitutes about 1/40th of the amount
grown in the country. In the United States and Canada, maize is mostly grown to
21
feed for livestock, as forage, silage (made by fermentation of chopped green
cornstalks), or grain. Maize meal is also a significant ingredient of some
commercial animal food products, such as dog food.
Alternative medicine
Stigmas from female maize flowers, popularly called corn silk, are sold as herbal
supplements.
Chemicals
Starch from maize can also be made into plastics, fabrics, adhesives, and many
other chemical products.
Bio-fuel
Feed maize" is being used increasingly for heating specialized corn stoves (similar
to wood stoves) are available and use either feed maize or wood pellets to
generate heat. Maize cobs are also used as a biomass fuel source. Maize is
relatively cheap and home-heating furnaces have been developed which use maize
kernels as a fuel. They feature a large hopper that feeds the uniformly sized maize
kernels (or wood pellets or cherry pits) into the fire.
Maize is increasingly used as a feedstock for the production of ethanol fuel.

Ethanol is mixed with gasoline to decrease the amount of pollutants emitted when
used to fuel motor vehicles. High fuel prices in mid-2007 led to higher demand
for ethanol, which in turn lead to higher prices paid to farmers for maize. This led
to the 2007 harvest being one of the most profitable maize crops in modern
history for farmers. Because of the relationship between fuel and maize, prices
paid for the crop now tend to track the price of oil.
The price of food is affected to a certain degree by the use of maize for biofuel
production. The cost of transportation, production, and marketing are a large
portion (80%) of the price of food in the United States. Higher energy costs affect
22
these costs, especially transportation. The increase in food prices the consumer
has been seeing is mainly due to the higher energy cost. The effect of biofuel
production on other food crop prices is indirect. Use of maize for biofuel
production increases the demand, and therefore price of maize. This, in turn,
results in farm acreage being diverted from other food crops to maize production.
This reduces the supply of the other food crops and increases their prices.
Fodder
Maize makes a greater quantity of epigeous mass than other cereal plants, so can
be used for fodder. Digestibility and palatability are higher when ensiled and
fermented, rather than dried.
Table 3.1 Gross chemical composition of different types of maize (%)
Crude Ether Carbo

Maize type Moisture Ash Protein
fibre extract hydrate
Sulpher 12.2 1.2 5.8 0.8 4.1 75.9
Crystalline 10.5 1.7 10.3 2.2 5.0 70.3
Floury 9.6 1.7 10.7 2.2 5.4 70.4
Starchy 11.2 2.9 9.1 1.8 22 72 8
Sweet 95 15 12.9 2.9 3.9 69.3
Pop 10.4 1.7 13.7 2.5 5.7 66.0
Black 12.3 1.2 5.2 1.0 4.4 75.9
Source: Cortez and Wild-Altamirano, 1972
3.2 Characteristics of sweet gourd

Cucurbita maxima, winter squash, pumpkin, calabaza, or marrow, is a species in
the gourd family (Cucurbitaceae) native to South America and cultivated by
indigenous people for over 2,000 years but now cultivated in warm areas
23
worldwide commercially and in home gardens as food and animal fodder and for
oil from the seeds. Cucurbita maxima, one of at least five species of cultivated
squash, is one of the most diverse domesticated species, perhaps with more
cultivated forms than any other crop. This species originated in South America
from the wild C. maxima ssp. andreana over 4000 years ago. The two species
hybridize quite readily but have noticeably different calcium levels. Different
squash types of this species were introduced into North America as early as the
16th century. By the American Revolution, the species was in cultivation by
Native American tribes throughout the present-day United States. By the early
19th century, at least three varieties are known to have been commercially
introduced in North America from seeds obtained from Native Americans.
Secondary centers of diversity include India, Bangladesh, Burma, and possibly the
southern Appalachians. C. maxima is the source of many varieties of winter
squash (including Acorn, Banana, Buttercup, Kabocha, Golden Delicious,
Hubbard, and Lakota), as well as giant pumpkin varieties (those that weigh over
45 kg [100 pounds]).

Kingdom: Plantae
Order: Cucurbitales
Family: Cucurbitaceae
Genus: Cucurbita
Species: C. maxima
Pumpkins are monoecious, having both male and female flowers on the same
plant. The female flower is distinguished by the small ovary at the base of the
petals. These bright and colorful flowers have extremely short life spans and may
only open for as short a time as one day.
24
Flower petal color : yellow .
Leaf type : the leaves are simple (lobed or unlobed but not separated into leaflets)
.
Leaf arrangement alternate: there is one leaf per node along the stem.
Flower symmetry : there are two or more ways to evenly divide the flower (the
flower is radially symmetrical). Number of sepals, petals or tepals there are five
petals, sepals, or tepals in the flower. Fusion of sepals and petals the petals or the
sepals are fused into a cup or tube.
Fruit type (general): the fruit is fleshy
Flower Sweet gourd Fruit
25
Table 3.2 Gross chemical composition of Sweet gourd
Principle Nutrient Value Percentage of RDA

Energy 26 Kcal 1%
Carbohydrates 6.50 g 5%
Protein 1.0 g 2%
Total Fat 0.1 g 0.5%
Cholesterol 0 mg 0%
Dietary Fiber 0.5 g 2%
Vitamins
Folates 16 mcg 4%
Niacin 0.600 mg 4%
Pantothenic acid 0.298 mg 6%
Pyridoxine 0.061 mg 5%
Riboflavin 0.110 mg 8.5%
Thiamin 0.050 mg 4%
Vitamin A 7384 IU 246%
Vitamin C 9.0 mg 15%
Vitamin E 1.06 mg 7%
Vitamin K 1.1 mcg 1%
Electrolytes
Sodium 1 mg 0.5%
Potassium 340 mg 7%
Minerals
Calcium 21 mg 2%
Copper 0.127 mg 14%
Iron 0.80 mg 10%
Magnesium 12 mg 3%
Manganese 0.125 mg 0.5%
Phosphorus 44mg 5%
Selenium 0.3 mcg <0.5%
Zinc 0.32 mg 3%
Phyto-nutrients
Carotene- 515 mcg --
Carotene- 3100 mcg --
Crypto-xanthin- 2145 mcg --
Lutein-zeaxanthin 1500 mcg --
Source: Okilo, 1984
26
3.3 Characteristics of sweet potato
The sweet potato (Ipomoea batatas) is a dicotyledonous plant that belongs to the
family Convolvulaceae. Its large, starchy, sweet-tasting, tuberous roots are a root
vegetable. The young leaves and shoots are sometimes eaten as greens. Ipomoea
batatas is native to the tropical regions in America. Of the approximately 50
genera and more than 1,000 species of Convolvulaceae, I. batatas is the only crop
plant of major importancesome others are used locally, but many are poisonous.
The sweet potato is only distantly related to the potato (Solanum tuberosum) and
does not belong to the nightshade family. The genus Ipomoea that contains the
sweet potato also includes several garden flowers called morning glories, though
that term is not usually extended to Ipomoea batatas. Some cultivars of Ipomoea
batatas are grown as ornamental plants; the name "tuberous morning glory" may
be used in a horticultural context.

Kingdom: Plantae
Order: Solanales
Family: Convolvulaceae
Genus: Ipomoea
Species: I. batatas
Sweet Potato is a tuberous-rooted perennial, usually grown as an annual. The top

is herbaceous, drying back to ground each year. The stems are forming a running
vine up to 4 m long, usually prostrate and slender, with milky juice, lateral stem-
branches arising from the short stem and usually not branched. The leaves are
ovate-cordate, borne on long petioles, palmately veined, angular or lobed,
depending on variety, green or purplish. The flowers are white or pale violet,
axillary, funnel-shaped, borne singly or in cymes on short peduncles. Fruits are
pods with 1-4 seeds per pod, flattened, hard-coated, angular.
27
Sweet potato leaf and flower Sweet potato
Table 3.3 Gross chemical composition of Sweet potato
Sweet potato, cooked, baked in skin, without salt

Nutritional value per 100 g (3.5 oz)
Energy 378 kJ (90 kcal)
Carbohydrates 20.7 g
Starch 7.05 g
Sugars 6.5 g
Dietary fiber 3.3 g
Fat 0.15 g
Protein 2.0 g
Source: Okilo, 1984
3.4 Ethanol Extraction
Ethanol extraction from the fermented material are estimated by different

methods. Some common methods of extraction are discussed below:
28
Types of ethanol extraction method are given below:
3.4.1 Water Bath Type Evaporator
The water bath type evaporator is a device for gently and efficiently evaporating
solvents from a mixture. It consists of two vessels (usually a large flask) and a
tube connecting them. One of the flasks contains the sample and the other holds
the extracted part; the tube attached to a condenser. The flask is heated by
emersion in hot water .Through heat transfer process; the contained liquid is
heated and boiled. Due to high temperature the solvent evaporates and passes
through the connecting tube and is condensed in the condenser section. The
condenser section is arranged so that the condensed vapors drain into another
flask where they are collected. It is a very efficient way and fast process for
extracting large quantities of solvent.
3.4.2 Rotary Evaporator
The rotary evaporator is a device for gently and efficiently evaporating solvents
from a mixture. It consists of a heated rotating vessel (usually a large flask) which
is maintained under a vacuum though a tube connecting it to a condenser. The
rotating flask is heated by partial emersion in a hot water bath. The flask's rotation
provides improved heat transfer to the contained liquid; the rotation also strongly
reduces the occupancies of 'bumps' caused by superheating of the liquid. The
solvent vapors leave the flask by the connecting tube and are condensed in the
condenser section. The condenser section is arranged so that the condensed vapors
drain into another flask where they are collected. It is a very efficient way but
comparatively slow process for removing large quantities of solvent.
3.4.3 Oil Bath Type Evaporator
The oil bath type evaporator is a device for gently and efficiently evaporating
solvents from a mixture. It consists of two vessels (usually a large flask) and a
tube connecting them. One of the flasks contains the sample and the other holds
the extracted part; the tube attached to a condenser. The flask is heated by
29
emersion in hot wax .Through heat transfer process; the contained liquid is heated
and boiled. Due to very high temperature the solvent evaporates and passes
through the connecting tube and is condensed in the condenser section. The
condenser section is arranged so that the condensed vapors drain into another
flask where they are collected. It is a very efficient way and fast process for
extracting large quantities of solvent.
3.4.4 Ethanol extraction
From all methods it had shown that the main process of extraction was
evaporating the ethanol from the solution. It is known that, at 78.3c ethanol is
evaporated from any solution. But in the experiment ethanol was mixed with raw
material and water as a form of Bio-ethanol. The evaporating temperature of
water is 100c, and it is essential to maintain a fix temperature for evaporating
only the Bio-ethanol from the mixer and that would be nearest to 80c. Because at
this temperature only ethanol will be evaporated from the mixer. Next the
evaporated Bio-ethanol will be collected and turned it into normal temperature.
This is the simple and profitable way to extract the Bio-ethanol from a raw mixer.
30
CHAPTER IV
METHODLOGY
This chapter deals with the methods employed for the research. Design of the
extraction process is simple.
4.1 Selection of sweet gourd, sweet potato and maize
Recent years production of sweet gourd, sweet potato and maize is increasing. So
the opportunity of producing Bio-ethanol is also increasing. Besides, demand for
Bio-ethanol is increasing day by day in Bangladesh.
Sweet gourd, sweet potato and maize contains high amount of sugar which is the
main reason for selecting those crops. The concept of producing Bio-ethanol from
different fruits, vegetables and root crops came from the notion that these have
considerable amounts of glucose, of the oligosaccharide group. Included in this
group are d-glucosamine, d-mannose, d-xylose, l-fucose in ratios of 2:2:1:1.
Exhaustive proteolysis resulted in the isolation of bromelains, glycopeptides
containing only Asx, Glx and Ser. Periodate oxidation, methylation, and
glycosidase digestion showed that the oligosaccharide chain has a highly branched
structure in which all the neutral sugars are in non-reducing terminal positions and
both N-acetyl-d-glucosamine residues occur in internal positions. This proves that
those can be a potential source of Bio-ethanol since the sugar found in its
cellulosic material is the chief raw material of Bio-ethanol production.
4.2 Study area
This experiment was conducted at Nilphamari Sadar, Nilphamari as shown in Fig

4.1. The experimental site was approximately situated between the North latitude
26019 and between North latitude 88 046 to 89 012 East longitude.
31
Fig 4.1 Study Area
32
4.3 Fermentation
Fermentation is a metabolic process that converts sugar to acids, gases and/or

alcohol. It occurs in yeast and bacteria, but also in oxygen-starved muscle cells, as
in the case of lactic acid fermentation. Fermentation is also used more broadly to
refer to the bulk growth of microorganisms on a growth medium. French
microbiologist Louis Pasteur is often remembered for his insights into
fermentation and its microbial causes. The science of fermentation is known as
zymology.
Fermentation takes place in the absence of oxygen (when the electron transport
chain is unusable) and becomes the cells primary means of ATP (energy)
production. It turns NADH and pyruvate produced in the glycolysis step into
NAD+ and various small molecules (see examples below). In the presence of O 2,
NADH and pyruvate are used in respiration; this is oxidative phosphorylation, it
generates a lot more ATP in addition to that created by glycolysis, and for that
reason cells generally benefit from avoiding fermentation when oxygen is
available. Exceptions include obligate anaerobes, which cannot tolerate oxygen.
The first step, glycolysis, is common to all fermentation pathways:
C6H12O6 + 2 NAD+ + 2 ADP + 2 Pi 2 CH3COCOO + 2 NADH + 2 ATP + 2
H2O + 2H+
Pyruvate is CH3COCOO. Pi is phosphate. Two ADP molecules and two Pi are
converted to two ATP and two water molecules via substrate-level
phosphorylation. Two molecules of NAD+ are also reduced to NADH.
In oxidative phosphorylation the energy for ATP formation is derived from an

electrochemical proton gradient generated across the inner mitochondrial
membrane (or, in the case of bacteria, the plasma membrane) via the electron
transport chain. Glycolysis has substrate-level phosphorylation (ATP generated
directly at the point of reaction)
33
4.3.1 Sweet gourd fermentation process
Fully mature ripe sweet gourds were selected. Sweet gourds were peeled off using
stainless steel knife. Then the sweet gourd was cut into pieces and 0.5kg of sweet
gourd and 500ml of water mixed in a fresh empty bottle. Then the mixer kept for
7 days in natural fermentation which is shown in Fig. 4.2
The method of fermentation of sweet Gourd shown below through flow diagram
Flow diagram of fermentation of sweet gourd
Selection of Sweet Gourd Only Ripe Sweet gourd were

selected
Peeled the sweet gourd Stainless steel knives were

used
Cut the sweet gourd Cut into pieces
Mixed with water By the help of hand
Fermentation Kept for fermentation for 7

days in plastic container
34
Fig 4.2 Sweet gourd fermentation
4.3.2 Sweet potato fermentation Process
Fully mature sweet potato was selected. Sweet potatoes were peeled off using
stainless steel knife. Then sweet potatoes were cut into pieces. Next 0.5kg of
sweet potato and 500ml of water mixed in a fresh empty bottle. Then the mixer
kept for 7 days in natural fermentation. Sweet potato fermentation was shown in
Fig. 4.3
The method of fermentation of sweet potato shown below through flow diagram
Flow diagram of fermentation of sweet potato
Selection of sweet potato Mature sweet potato were

selected
Knives were used

Peeled the sweet potato
Cut the sweet potato Cut into pieces
Mixed with water By the help of hand
Fermentation Kept for fermentation for 7

days in plastic container
35
Fig 4.3 Sweet potato fermentation
4.3.3 Maize fermentation Process
Ripe maize was selected. The grain was removed from total maize. Then it was
cut into pieces and boiled for 15 minutes and 0.5kg of maize and 500ml of water
mixed in a fresh empty bottle. Then the mixer kept for 7 days in natural
fermentation was shown in Fig 4.4
The method of fermentation of Maize shown below through flow diagram
Flow diagram of fermentation of maize
Selection of maize Ripe maizes were selected
Removed maize grain Steel knives were used
By the help of Hand

Mixed with water
Kept for the fermentation for 7

Fermentation days in a plastic container
36
Fig 4.4 Maize fermentation
Table 4.1: Quantity of total fermentation
Sl Duration of
Crop type Amount Water
no: fermentation
1. Sweet gourd 500 mg 500 ml 7 days
2. Sweet potato 500 mg 500 ml 7 days
3. Maize 500 mg 500 ml 7 days
4.4 Total fermentation
Total fermentation process of sweet gourd, sweet potato and maize for 7 days is
shown in Fig 4.5
37
Fig 4.5 Quantity of total fermentation
4.5 Process of extraction
After 7 days of natural fermentation, 500ml of sweet gourd, sweet potato and
maize were collected in 3 separate bottles shown in Fig 4.6. A simple evaporating
system was made for extracting the Bio-ethanol from raw materials. In the
evaporator an electric heater was used which shown in Fig 4.7, which control
temperature. The heater was connected with a bottle which was saturated in
normal temperature water for cooling.
Fig 4.6 Before extraction
38
Fig 4.7 Evaporation system by using heater
4.6 Collection of Bio-ethanol

Finally the Bio-ethanol was collected in three separate bottles for further chemical
analysis, which was shown in Fig. 4.8
Fig 4.8 Collection of Bio-ethanol
39
CHAPTER V
RESULTS AND DISCUSSION
This chapter comprises of the discussion of the results obtained from the
experiment conducted.
5.1 Ethanol from Sweet gourd
In the first sample 2 kg sweet gourd was collected from the market. After peeling
sweet gourd, the weight was 1.75 kg. Then the sweet gourd smashed and mixed
with 500ml of fresh water without any yeast. After 7 days fermentation 500ml raw
material was collected from the fermentation for extraction. Then the raw material
was put in the evaporate chamber to separate the Bio-ethanol from the solution.
The Bio-ethanol was collected after 2 hours evaporation. The amount of collected
Bio-ethanol was 95ml. Now all the results are given below. These are shown in
Table 5.1.
Table 5.1 Sweet gourd fermentation
Volume of
Name of Initial weight Weight after Volume of raw
Bio-ethanol
sample (kg) peeling ( kg) material (ml)
(ml)
Sweet gourd 2 1.75 500 95
Table 5.2 Ethanol Extraction in percentage
No of sample Amount of sample Amount of Bio- Percent of Bio-

(ml) ethanol (ml) ethanol (%)
Sweet Gourd 500 95 19.00
Result 95 19
5.2 Ethanol from sweet potato
In first sample 2 kg potato was collected from market. After peeling sweet potato,
the weight was 1.8 kg. After cutting all sweet potatoes into small pieces and sweet
potato was mixed with 500ml of fresh water without any yeast. After 7 days
fermentation 500ml raw materials were collected for extraction. Then the raw
40
materials was put in the evaporate chamber to separating the Bio-ethanol from the
solution. After 2.5 hours evaporation the Bio-ethanol was collected. The amount
of collection of Bio-ethanol was 105ml. Now all the results are given below.
These are shown in Table 5.3.
Table 5.3 Sweet potato fermentation
Volume of
Volume of raw
Name of Initial weight Weight after Bio-
material
sample (kg) peeling ( kg) ethanol
(ml)
(ml)
Sweet
2 1.80 500 105
potato
Amount of sample Amount of ethanol Percent of

No of sample
(ml) ( ml) ethanol
Sweet potato 500 105 21.00
Result 105 21.00
5.3 Ethanol from maize
In first sample 2 kg maize was collected from market. Here the entire sample was
used for fermentation. Collected maizes were partially smashed and mixed with
500ml of fresh water without any yeast. After 7 days fermentation 500ml raw
material was collected to extraction. Then the raw material was put in the
evaporate chamber for separating the Bio-ethanol from the solution. After 2.5
hours evaporation the Bio-ethanol was collected. The amount of collected of Bio-
ethanol is 110ml. Results are shown in Table 5.5
Table 5.5 Ethanol from maize
Name of Initial weight Weight after Volume of raw Volume of

sample (kg) peeling ( kg) material Bio-
(ml) ethanol
(ml)
Maize 2 2.00 500 110
41
Amount of sample Amount of Bio- Percent of Bio-

No of sample
(ml) ethanol (ml) ethanol (%)
M 500 110 22.00
Result 110 22.00
5.4 Comparison of percentages of different crops extracted Bio-ethanol

According to the experiment it was found that, the percentage of sweet gourd,
sweet potato and maize are 19%, 21% and 22% respectively. Comparison of
percentage and price showed in Fig 5.1
Fig 5.1 Comparison of percentages and prices of different crops extracted

Bio-ethanol
Table 5.7 According to market price of those crops and its Bio-ethanol
percentage
Sl. Price in tk. Bio-ethanol

Crop name
no (1kg) percentage
1 Sweet gourd 10 19
2 Sweet potato 10 21
3 Maize 12 22
42
5.5 Extraction process of Bio-ethanol
According to the experiment and other extraction methods, it is the easy and
profitable method to extract the Bio-ethanol from raw material.
Fig 5.2 Bio-ethanol extraction process:
Bio-Ethanol Extraction Process
Sugar Platform
Sweet Gourd Extraction Fermentation Beer

Sweet Potato Sugars (~15% EtOH)
Maize
Distillation
> 90% Ethanol
Dehydration
> 99% Ethanol
Fig 5.2 Bio-Ethanol extraction process
5.6 Chemical conversion process of Bio-ethanol
In hydrolysis process firstly starch converted to glucose then by fermentation

process glucose convert into ethanol for the presence of microorganisms.
43
Fig 5.3 Starch to ethanol process from Raw material:
Starch to Ethanol Process
Hydrolysis
(C6H10O5)n + n H2O Anaylases n C6H12O6
Fermentation
C6H12O6 2C2H5OH + 2CO2
Fig 5.3 Starch to Ethanol process
5.7 Chemical analysis of Bio-ethanol
Raw materials were sent to Rajshahi University chemistry lab for chemical analysis
of Bio-ethanol.
Table 5.8 Chemical analysis
Percentage of
Sl. Percentage of Percentage of
Crop name other materials
no ethanol (%) water (%)
(%)
01 Sweet Gourd 46 34 20
02 Sweet potato 32 47 21
03 Maize 60 28 12
44
5.8 Production cost of Extracted bio-ethanol
Fixed cost:
Table 5.9: Fixed cost for conducting experiment (For 3 years)

Sample Materials Amount Cost (Tk.)
no.
1. Heater 1 2250
2. Plastic pipe 3.5 ft 140
3. Plastic bottle 7 pieces 150
4. Plastic pot 2 pieces 240
5. Smasher 1 pieces 100
6. Knife 1 pieces 30
Total cost 2910 (Tk.)
After 3 years later the value of those materials will be 1000Tk for depreciation. So
the fixed cost will be= (2910-1000)/ (312) =53Tk/month.
Variable Cost:
Sweet gourd/opportunity cost=3000Tk(10 Tk/kg)
Labor cost = 300Tk
Transport fee (Total) =100Tk
Electric bill (Total)=750Tk
Total cost (Sweet gourd) = (Fixed Cost + Variable cost)

= (53Tk/month + Tk. (3000+300+100+750)/month)
=53Tk/month + 4150Tk/month
Total Income (Per month)
Bio-ethanol amount =75 liter
Income from Bio-ethanol= 4500Tk
By product amount= 90 kg
Income from bi-product = 450Tk
45
Total income = (4500+450)Tk = 4950Tk
Net benefit:
Net benefit=Total income-Total cost
= (4950-4150)Tk
= 800Tk
Cost-benefit ratio:
Yearly total cost= 49800Tk
Yearly total income = 59400Tk
Benefit cost ratio =
= 1.193
Total calculation was conducted according to the amount of 300 kg of Sweet Gourd,
Sweet Potato and Maize. For each ton of raw material this procedure brings
32000Tk/ton, 33666Tk/ton and 35000Tk/ton respectively for of Sweet Gourd Sweet
Potato and Maize.
5.9 Price of Bio-ethanol

Bio-ethanol mainly used in European country as a fuel at $80-90 per barrel. That
means 60Tk per liter in our country. From that aspect the analysis is given below.
Table 5.10: Price of Bio-ethanol
Variable Total Benefit

Sample Fixed cost Total cost
Element cost Income cost
No. (Tk./year) (Tk./year)
(Tk./year) (Tk./yr) ratio
01 Sweet gourd 636 49164 49800 59400 1.193
02 Sweet potato 636 49164 49800 59909 1.203
03 Maize 636 52764 53400 63919.8 1.197
46
CHAPTER VI
CONCLUSIONS AND RECOMMENDATIONS
6.1 Conclusions
In Bangladesh there is a great opportunity to develop the market of raw bio-

ethanol for producing ethanol. In this experiment it was shown that how the bio-
ethanol will be collected. Nowadays, in the north zone of our country the
production of maize and sweet gourd is increasing very rapidly. In the same time
the use of fuel in our regular life is also increasing. So it is needed to fill-up the
requirement of fuel and bio-fuel may be the way to fill-up the requirement.
Bio-ethanol is seen as a good fuel alternative because the source crops can be
grown renewably and in most climates around the world. In addition the use of
Bio-ethanol is generally CO2 neutral. This is achieved because in the growing
phase of the source crop, CO2 is absorbed by the plant and oxygen is released in
the same volume that CO 2 is produced in the combustion of the fuel. This creates
an obvious advantage over fossil fuels which only emit CO 2 as well as other
poisonous emissions.
Bio-fuel is an upcoming renewable source of energy, which would not only help
in controlling the demand, but also will reduce pollution. If we can select the most
efficient source of ethanol production then the cost will be reduced. Energy sector
will be more profitable which can play an important role in the extension of fuel
business. This extension of fuel business is necessary for fulfill the shortage of
fuel energy and offers sound environment.
There is a global oil crisis, as the demand for petroleum increases each year while
our supplies are rapidly being depleted. Bio-ethanol, a principle bio-fuel, is a
natural alternative to gasoline. One solution is to produce bio-ethanol from
different nontraditional and traditional fruits, vegetables and root crops.
47
This study conducted the extraction of bio-ethanol from Sweet gourd, Sweet
potato and Maize through fermentation without using the yeast will indeed be a
great help to the economy and the environment. Sweet gourd, Sweet potato and
Maize are suitable for Bio-ethanol production. The highest concentration of
ethanol has been observed in Maize about 60%, others were 46% and 32% ethanol
from sweet gourd, sweet potato and maize respectively in this experiment. These
results state that the Maize is a potential raw material for bio-ethanol extraction
.This research will also answer the crisis of looking for a clean, alternative source
of fuel energy. In this mechanical system obtained bio-ethanol from 500 ml sweet
gourd, sweet potato and maize solution are 95 ml, 105 ml and 110 ml only but
there need to invest huge amount of money. It was found from the calculation
that the developed system can bring 32000Tk/ton, 33666Tk/ton and 35000Tk/ton
with benefit-cost ratio 1.193, 1.207 and 1.197 respectively. These research will
also solution the crisis of looking for a clean, alternative source of fuel energy.
6.2 Recommendations:
1. Evaporating extraction method is profitable.
2. If the extraction of Bio-ethanol will be done in large scale than it will be more
profitable.
3. For proper extraction the temperature of the heater should be maintained

properly.
4. Further research on the quality of Bio-ethanol should be conducted.
48
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56
LIST OF ABBREVIATION
BINA : Bangladesh Institute of Nuclear Agriculture

BRRI : Bangladesh Rice Research Institute
BAU : Bangladesh Agricultural University
Contd. : Continued
et.al : et alia (and others)
FAO : Food and Agriculture Organizations of the United Nations
ha : Hectare
M.ton : Metric ton
No. : Number
Mha : Million hectare
ml : Milliliter
RU : Rajshahi University
DAE : Department of Agricultural Extension
o
C : Degree Celsius
% : Percentage

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STUDY ON BIO-ETHANOL EXTRACTION PROCEDURE

FROM SWEET GOURD, SWEET POTATO AND MAIZE

HUSSAIN MOHAMMAD KHALEDUJJAMAN

DEPARTMENT OF FARM POWER AND MACHIENARY

HUSSAIN MOHAMMAD KHALEDUJJAMAN

In partial fulfillment of the requirement for the

DEPARTMENT OF FARM POWER AND MACHIENARY

HUSSAIN MOHAMMAD KHALEDUJJAMAN

Approved as to style and content by

DEPARTMENT OF FARM POWER AND MACHIENARY

Chapter No. Title Page No.

Chapter No. Title Page No.

3.4.4 Ethanol extraction 30

Table No. Title Page No.

Figure No. Title Page No.

4.1 Study Area 32

This experiment was conducted to bring in a suitable and economic bio-ethanol

The author expresses profound appreciation, heartfelt gratitude and immense

As demand for the limited global supply of nonrenewable energy resources

Bioethanol is a microbiological way of converting simple sugar into ethanol and

(1 lakh (1 lakh (1 lakh (1 lakh (1 lakh (1 lakh (1 lakh (1 lakh (1 lakh

Source: Agriculture Diary 2014 and report of DAE

1.2 Specific objectives are:

2.1 Raw Materials for Bio-ethanol Production

Maputo Mozambican (2007) scientists and researchers told an International

2.2 Bio- ethanol Production Methods

Islam (2009) conducted an experiment on mechanical extraction method (

Avril Rodiel Bries (2008), submitted an investigatory project report in 16 TH

Teng-WenYuh et al. ( 2005) reported the efficiency of carbon dioxide

Lautenschlaeger. Friendrich k (2004) studied the treatment of fruit, vegetable ,

2.3 Bio- ethanol Extraction From Different Sources

In recent years, however, research is focused on processes involving amylolytic

2.4 Effect on Environment

CHARACTERISTICS OF MAIZE, SWEET GOURD AND SWEET

3.1 Characteristics of maize

3.1.1 Distribution and habit

Maize, known in some English-speaking countries as corn, is a large grain plant

3.1.2 Botanical Feature

Corn tassel Corn silks Corn ear Ripened corn

Maize is increasingly used as a feedstock for the production of ethanol fuel.

Table 3.1 Gross chemical composition of different types of maize (%)

Crude Ether Carbo

Sulpher 12.2 1.2 5.8 0.8 4.1 75.9

Crystalline 10.5 1.7 10.3 2.2 5.0 70.3

Floury 9.6 1.7 10.7 2.2 5.4 70.4

Starchy 11.2 2.9 9.1 1.8 22 72 8

Sweet 95 15 12.9 2.9 3.9 69.3

Pop 10.4 1.7 13.7 2.5 5.7 66.0

Black 12.3 1.2 5.2 1.0 4.4 75.9

Source: Cortez and Wild-Altamirano, 1972

3.2 Characteristics of sweet gourd

3.2.2 Botanical Feature

Fruit type (general): the fruit is fleshy

Flower Sweet gourd Fruit

Principle Nutrient Value Percentage of RDA

Source: Okilo, 1984

3.3.2 Botanical Feature

Sweet Potato is a tuberous-rooted perennial, usually grown as an annual. The top

Table 3.3 Gross chemical composition of Sweet potato

Sweet potato, cooked, baked in skin, without salt

Source: Okilo, 1984

3.4 Ethanol Extraction

Ethanol extraction from the fermented material are estimated by different