Preparation of Ethyl Alcohol by Fermentation of Molasses and Alternative Raw

Materials

A Research Study Presented to the

Faculty of the Department of Chemical Engineering

School of Engineering and Architecture

Saint Louis University

In Partial Fulfillment of the Requirements for the Degree

Bachelor of Science in Chemical Engineering

By

Students

September 2018

i
ACKNOWLEDGEMENT

The researchers of this experiment would like to thank first our Almighty God for the

benisons and wisdom that gave the researchers enough strength, knowledge and attitude to do

and finish this experiment accurately and safely.

The researchers are also very much thankful for the guidance and knowledge that

their instructor, Engr. X , have imparted to them which helped a lot in the finalization of the

papers and understanding of the procedures. Also, the SLU administration for lending the

researchers the complete equipment to be used in the experiment.

The researchers also like to extend their appreciation to those who are not mentioned

here but somehow played a purpose on this activity.

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TABLE OF CONTENTS

TITLEPAGE i

ACKNOWLEDGEMENT ii

TABLEOFCONTENTS iii

LIST OF TABLES iv

ABSTRACT v

Chapter 1: INTRODUCTION 1

Chapter 2: DESIGN AND METHODOLOGY 5

Chapter 3: RESULTS AND DISCUSSION 7

Chapter 4: CONCLUSION AND RECOMMENDATION 8

REFERENCES 9

APPENDICES

a. List and uses of Apparatus 10

b. Definition of Terms 13

c. Problems 14

d. Pictorials 15

iii
ABSTRACT

VISCOSITY IS THE RESISTANCE OF A FLUID TO FLOW, IT RESULTS FROM

INTERMOLECULAR ATTRACTIONS THAT IMPEDE THE MOVEMENT OF

MOLECULES AROUND AND PAST EACH OTHER. THE EXPERIMENT WAS

CONDUCTED TO DETERMINE THE ABSOLUTE VISCOSITY OF ORGANIC

LIQUIDS AND THE RELATIONSHIP OF VISCOSITY AND DENSITY USING

OSTWALD VISCOMETER. USING OSTWALD VISCOMETER, A DEFINITE

VOLUME OF THE LIQUID WAS PLACED. THE LIQUID WAS ALLOWED TO

FLOW OUT PASSING TWO CALIBRATED MARKS, AND THE FLOW TIME WAS

MEASURED. THE DENSITIES WERE CALCULATED USING THE CONSTANTS

AND FORMULA FROM THE HANDBOOK. THE VISCOSITIES OF THE LIQUIDS

WERE CALCULATED BY APPLYING THE POISEUILLE'S LAW, WHEREIN

VISCOSITY IS DIRECTLY PROPORTIONAL TO THE SAMPLE'S DENSITY,

FLOW TIME, AND THE VISCOSITY COEFFICIENT. TO DETERMINE THE

VISCOSITY COEFFICIENT, THE VISCOMETER WAS CALIBRATED USING

DISTILLED WATER AS THE REFERENCE LIQUID. RESULTS FOR THE

VISCOSITIES OF ORGANIC LIQUIDS (BENZENE, CHLOROFORM, METHANOL

AND THE UNKNOWN) WERE 5.956x10-4 Pa•s, 6.4579x10-4 Pa•s, 6.8301x10-4 Pa•s ,

and 5.4631x10-4 Pa•s RESPECTIVELY. COMPARING THESE VALUES TO THE

TRUE VALUES OF, IT WAS NOTICED THAT SOME WERE QUITE DIFFERENT.

HOWEVER, IT CAN BE CONCLUDED THAT USING OSTWALD VISCOMETER

IS AN EFFECTIVE WAY OF MEASURING ABSOLUTE VISCOSITY OF LIQUIDS.

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Keywords: Dumas Method, Ideal Gas Equation, molar mass

v
Chapter 1

INTRODUCTION

Fermentation is a chemical process by which molecules such as glucose are broken

down anaerobically and it is foaming that occurs during the manufacture of wine and beer.

Fermentation in a narrow sense to describe the changes brought about by yeasts and other

microorganisms growing in the absence of air. Ethyl alcohol and carbon dioxide are not the

only products of fermentation. Ethanol known as ethyl alcohol or grain alcohol is a flammable,

colorless, mildly toxic chemical compound with a distinctive perfume –like odor, and the

ethanol is found in alcoholic beverages. Natural energy resources such as petroleum and coal

have been consumed at high rates over the last decades. The heavy reliance of the modern

economy on these fuels is bound to end due to their environmental impact and to the fact that

they might eventually run out. Therefore, alternative resources such as ethanol are becoming

more important. Bio-ethanol is one of the most important renewable fuels contributing to the

reduction of negative environmental impacts generated by the worldwide utilization of the

fossil fuels. It is also used a solvent in many fields and used as a motor fuel additive.

Ethanol is prepared by molasses fermentation. A residue begotten from sugar cane

processing, molasses is the mother liquor left after crystallization of sugarcane juice. It is a

dark colored viscous liquid that contains approximately 40 to 50% fermentable sugar. Being

one of the earliest biotechnologies used by humans for the production of Ethanol, molasses

Fermentation is also the most cost-effective way. Molasses is a microbiological energy source

that helps in the process of growing yeast, bacteria and molds. Not only is it the most cost-

1
effective amongst the available energy sources for such industries but is also the easiest to

incorporate in fermentation processes.

Molasses Fermentation is a biological process in which sucrose from molasses is

converted into cellular energy that eventually produces ethanol and carbon dioxide. Molasses

from sugarcane or sugar imparts an appropriate substrate for ethanol production. They are the

basic sources of sucrose which is present in them in concentrated sugar form. Fermentable

sugars are directly used during Molasses fermentation and with the help of enzymes and

reactions ethanol is formed. The Ethanol begotten from the whole process is used in the form

of absolute or rectified spirit.

. In the fermentation of ethyl alcohol you can use many alternative raw materials like

fruit juice such as orange juice. Orange are dominant in our country and the over ripped oranges

can be converted to very useful product which is the ethanol. The use of the fruit as a substrate

for producing high added value products has been accomplished an example is spirits obtained

by the fermentation and distillation of fruit . Fruit spirits are produced all over the world using

various fruits, according to the availability in different countries and seasons. In this way, the

current commercialization of known alcoholic beverages obtained from fruit could facilitate

the market penetration of such spirits

The process needed to produce fruit spirit is complex and involves various factors that

influence the quality of the final product. However, the main physico‐chemical and sensorial

differences among spirits are due to the particular composition of their corresponding raw

materials like fruit, cereals, vegetables, etc and the fermentation process. Market‐orientated

yeast strains are currently being developed for the competitive production of alcoholic

beverages with minimized resource inputs, improved quality and low environmental impacts .

2
The orange juice, the initial sugar concentration was approximately 11ºBrix, and the pH was

3.7. The orange juice was mixed with a sucrose solution to adjust the sugar concentration to

16°Brix. Using some alternative raw materials in the production of ethanol is very essential

and helpful to environment to minimize the use of resources.

of organic liquids using the Ostwald viscometer. In order to achieve the goal, the following

questions must be answered: (1) What is the flow time of the liquid, (2) what is the density of

the liquid, (3) what is the temperature at the given density, and (4) what is the viscosity of the

liquid? The measure of the viscosity in this experiment is the drag caused by the relative motion

of the fluid and the surface while the flow conditions must have a sufficiently small value of

Reynolds number for there to be a laminar flow. The Ostwald viscometer fulfils these

conditions: A U-tube with two reservoir bulbs separated by a capillary. The liquid is added to

the viscometer, pulled into the upper reservoir by suction, and then allowed to drain by gravity

back into the lower reservoir. The time that it takes for the liquid to pass between two etched

marks, one above and one bellow the upper reservoir, is measured. For absolute measurement

we have to know all parameters of the viscometer (Viscosity coefficient) but we can calibrate

the equipment using a reference liquid having well known density like water. Knowing the

viscosity of the reference liquid, the viscosity of the liquid can be calculated.

In doing this experiment, it is important that the students know beforehand the basic

principle that the density is the main factor that brings about the differences in the viscosity of

different liquids. The viscosity of the different liquids can be calculated from the constants and

the formula given in the Perry's Chemical Engineers' Handbook. Also, viscosity varies with

temperature. In general, the viscosity of a sample liquid decreases with increasing temperature

and vice versa. Viscosity is normally independent of pressure. Since liquids are normally

3
incompressible, an increase in pressure doesn't really bring the molecules significantly closer

together. Knowing the viscosity of a sample is important because it plays a major role to our

daily lives. In the field of Chemical Engineering, viscosity measurement plays a big part in the

industry. Viscosity is an important fluid property when analyzing liquid behavior and fluid

4
Chapter 2

DESIGN AND METHODOLOGY

This study focuses on preparing ethyl alcohol by fermentation of locally available

materials like molasses. And determining or confirming the presence of ethyl alcohol in the

fermentation mixture by standardization. The materials used are graduated cylinder, 500-ml

round bottom flask, Erlenmeyer flask, beakers, stopper with bent tube, distilling flask, water

condenser with rubber tubing, test tubes, watch glass, Pasteur pipette/dropper, iron stand, iron

clamps, Bunsen burner and cotton. The reagents used are Molasses, distilled water, Pasteur’s

nutrient and limewater.

During the experiment 50 grams of Molasses was dissolved in 250 ml of distilled

water placed in a 500 ml round bottom flask. When the molasses was thoroughly dissolved, 50

ml Pasteur’s nutrient was added. The mixture was shake thoroughly then the flask was closed

with a stopper, fitted with bent tubes. The other side of the tube was attached to a test tube

filled with 12 ml of limewater then it was sealed at the top with a cotton wad. The mixture was

left in the locker for a week at room temperature.

After a week, the fermentation process was completed. Without disturbing the

mixture, it was the decanted and placed in a distilling flask attached to a water condenser. The

mixture was then distilled, as soon as 2ml of sample was collected it was subjected to a

combustion test then the next 5ml of sample collected was subjected to an iodoform test.

In the combustion test, the 2 ml sample was placed in a watch glass and was ignited.

While in the iodoform test, the 5 ml of KI solution was added to the 5 ml sample then drop by

5
drop KOH solution was added. It was set aside for an hour to wait for the formation of

iodoform.

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Chapter 3

RESULTS AND DISCUSSION

1. Observation for Combustion test

The results of using both the molasses and the alternative raw material to prepare

ethyl alcohol showed positive results in the combustion test. After being in contact

with the flame of the match the sample distillate of both the molasses and the

alternative raw material lit. The flame produce was bluish in colour with a tinge of

orange. It can be said that both are combustible and volatile in nature.

2. Observation for Iodoform test

Even after adding 5 ml of the solution iodine in KI and then with KOH solution drop

by drop, since the distillate is with no color originally it was not decolorized in a

sense because in the first place it was colorless. After setting the mixture for an hour,

there were no formations of the iodoform.

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Chapter 4

CONCLUSION AND RECOMMENDATION

Fermentation is a chemical process by which molecules such as glucose are broken

down anaerobically and it is foaming that occurs during the manufacture of wine and beer.

In the experiment done, it was proven that it is possible to prepare ethyl alcohol by

fermentation of locally available raw materials like molasses and orange juice, in our case.

The results of using both the molasses and the alternative raw material to prepare ethyl

alcohol showed positive results in the combustion test. After pouring 2 ml of both sample

distillates onto an evaporating dish, the sample distillate of both the molasses and the

alternative raw material lit upon contact with the flame of the match. It can be said that both

are combustible and volatile in nature.

On the other hand, after performing the iodoform test it showed no reactions, even after

adding 5 ml of the solution iodine in KI and then with KOH solution drop by drop. The

distillate remained colorless and after setting the mixture for an hour, there were no formations

of the iodoform.

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References:

Laboratory Test and Notebook for Organic Chemistry by Bossert, Roy and Wallace KL.
Brode, 4th edition.

Module on Science and Technology Education in the Phillipine Society by Tan, Merle C.

Laboratory Courses in Organic Chemistry by Rosenblatt, David H.Laboratory

Manual of Elementary Organic Chemistry by Richer, Ross.

https://en.wujipedia.org/wiki/ethanol.

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APPENDIX A

LIST AND USES OF APPARATUS

 Graduated cylinder - is a common piece of laboratory

equipment used to measure the volume of a liquid. It has a
narrow cylindrical shape. Each marked line on the graduated
cylinder represents the amount of liquid that has been
measured.

 Round bottom flask- are often used to contain chemical

reactions run by chemists, especially for reflux set-ups and
laboratory-scale synthesis. This flask shape is also more
resistant to fracturing under vacuum as a sphere more evenly
distributes stress across its surface.

 Erlenmeyer flask- its conical shape and narrow neck are its
distinguishing features. Easy pouring and long term storage when
used with a stopper reducing the evaporation of stored liquids &
limiting contamination by airborne bacterial, fungal and chemical
contaminants.

 Beaker- is a cylindrical container used to store, mix and heat

liquids in laboratories. Most are made of glass but other non-
corrosive materials such as metal and heat-resistant plastics. It
is usually have a flat bottoms and a lip around the top.

 Stopper with stopper - is a truncated cylindrical or conical

closure to seal a container such as a bottle, tube or barrel. It
encloses a container from the outside without displacing the
inner volume

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 Distilling flask- is a round bottom and a long neck from
which a side arm protrudes that is used to separate mixtures of
two liquids with different boiling points

 Water condenser- is a device used in the laboratory to cool

hot gases into liquids. It is usually a long,
circular glass tube inside the tube there is another
smaller tube. The hot gas goes through the smaller
inside tube while in the outside one
cold water goes around. Water can be put in from
the tap, and goes from the bottom to exit at the top.

 Test tubes- are usually made of glass for better resistance to

heat and corrosive chemicals. It is used to handle chemicals
especially for qualitative since their spherical bottom and
vertical sides reduce mass loss when pouring, make them
easier to wash out

 Watch glass- circular concave piece of glass used in

chemistry as a surface to evaporate a liquid, to hold solids
while being weighed for heating a small amount of
substance and as a cover for a beaker. It is also used as an
evaporation surface, a watch glass allows closer observation
of precipitates or crystallization, and can be placed on a surface of contrasting color
to improve the visibility overall.

 Pasteur dropper- is also known as droppers or eye droppers

used to transfer small quantities of liquids. They are
usually glass tubes tapered to a narrow point, and fitted with
a rubber bulb at the top.

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 Iron clamp – is a piece of laboratory equipment used to
hold things and is placed in an iron stand to elevate and
hold the equipment

 Iron stand- is a metal rod attached to heavy metal base and keeps the
stand stable and vertical metal rod allows for easy height adjustment
of the iron ring/clamp
 Bunsen burner- device for combining a flammable gas with
controlled amounts of air before ignition; it produces a
hotter flame than would be possible using the ambient air and gas
alone. It provides heat during the experiment.

 Cottons – it is used as bung or stopper instead of

a rubber bung because the yeast-glucose mixture
produces carbon dioxide as the fermentation
process takes place which could cause a rubber
bung to become dislodged.

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APPENDIX B

DEFINITION OF TERMS

o Fermentation- is a metabolic process that consumes sugar in the absence of oxygen.

The products are organic acids, gases, or alcohol. It occurs in yeast and bacteria.

o Ethyl alcohol - is the alcohol found in alcoholic drinks such as beer, brandy, or
whiskey. It is made from the fermentation or chemical breakdown of sugars by yeast

o Molasses –black treacle, is a viscous product resulting from refining sugarcane or

sugar beets into sugar. Molasses varies by amount of sugar, method of extraction, and
age of plant.

o Combustion test- a test performed wherein a sample liquid is to be ignited to find out
certain substances compsing the sample.

o Iodoform test- The reaction of iodine and base with methyl ketones is so reliable that
the "iodoform test" (the appearance of a yellow precipitate) is used to probe the
presence of a methyl ketone. This is also the case when testing for specific secondary
alcohols containing at least one methyl group in alpha-position.

o Nutrient salt solution – provides the essential mineral and nitrogen sources for the
growing yeast cells so that they in turn can provide the enzyme zymase

o Zymase - a mixture of enzymes obtained from yeast that catalyze the breakdown of
sugars in alcoholic fermentation.

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APPENDIX C

PROBLEMS

1. Define fermentation. Discuss the process of fermentation.

Fermentation is a metabolic process in which an organism converts a carbohydrate,
such as starch or a sugar, into an alcohol or an acid. For example, yeast performs
fermentation to obtain energy by converting sugar into alcohol. In the experiment
done molasses was used as the sugar substituent. Bacteria perform fermentation,
converting carbohydrates into lactic acid.

2. Give the complete reactions in the fermentation of molasses

C12H22O11 +H2O C6H12O6 +C6H12O6

C6H12O6 C2H5OH + 2CO2

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APPENDIX D

PICTORIALS

The picture above illustrates the set-up

done for the fermentation process of
molasses after it was dissolved and
prepared with the Pasteur’s nutrient
solution After a week of fermenting, the
fermented molasses-ethyl
alcohol is ready for distilling

The solution in the

picture above was
then distilled using
the set-up shown in
the image in the
left. Using a
fractionating
column, the disllate
was then used for
other tests in the
experiment

15
Two ml of the distillate was measured and was then placed in an evaporating
dish. It was then ignited for the combustion test part of the experiment.

The raw material that was used in the experiment was orange. It was
peeled, cut into smaller pieces and was then blended in the blender

16
The image above was the The alternative raw
product after it was material was then
blended and placed in a placed in a clean
thin cloth in order to container ready for
extract the juice diistillation

17
After performing the
same steps as the
procedure done in the
molasses
fermentation, the
fermented orange
juice was then filtered
in order for the pulps
and residues to be
discared and not be
included in the
distilling process

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After fermenting the orange juice, it was then distilled in order to get
the final product.

Combustion test
was performed with
2 ml of distillate to
determine the
presence of alcohol
in the product

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