K.SWARNA LATHA.

(Y15CH819)
M.DILEEP. (L16CH868)
G.S.N.D.SURESH. (L16CH871)

Under guidance of
Mrs. J. L. JAYANTHI
(M.Tech)
ABSTRACT:-
Production of Ethanol fermented from renewable sources for fuel or fuel additives are
known as bio-ethanol.
Large quantities of fruit waste are generated from agricultural processes worldwide.
This waste is often simply dumped into landfills or the ocean. Fruit waste has high
levels of sugars, including sucrose, glucose, and fructose, that can be fermented for bio-
ethanol production.
the need of bio-ethanol has been increasing, the production of bio-ethanol must be
increased using cheaper and eco friendly raw materials. On the basis of these
characteristics fruit wastes can be considered as cheaper and eco friendly.
 In this study different fruit wastes were used as a raw material for the production of
bio-ethanol by using Saccharomyces cerevisiae.
INTRODUCTION:-

The largest single use of ethanol is as a motor fuel and fuel additive.
Alcohol is an organic compound that has one or more hydroxyl (OH) groups
attached to a carbon atom. In dilute aqueous solution, it has a somewhat sweet
flavor, but in more concentrated solutions it has a burning taste.
The first generation of ethanol production used corn as a substrate, later corn was
considered as a feedstock lead to the second generation of production of ethanol
which used microorganisms and different wastes as substrates.
The cheapest and easily available source for the production of bio-ethanol is fruit
wastes.
It is a potential energy source, from which ethanol can be obtained. Fruit waste.
OBJECTIVES:-

 Preparation of bio-ethanol by different fruit wastes.

Compare alcohol percentage obtained from different fruit wastes.

Calculating the calorific value and concentration of bio-ethanol.
Compare the ethanol properties with different journal results.
APPLICATIONS:-
As a transport fuel to replace gasoline.
Pharmaceutical industries.
In chemical industries.
To prepare solvent based paint and thinners.
In beauty care industry (perfumes ,hair gel etc)
ADVANTAGES:-
 It reduces greenhouse gases through the use of bio-ethanol blended fuel.
The fuel spills are more biodegradable or diluted to non toxic concentrations.
The exhaust gases of ethanol are much cleaner. It undergoes complete combustion.
Bio-ethanol is also biodegradable and less toxic then fossil fuels.
Process cost is very less
Raw materials are easily available and production process is simple.
DISADVANTAGES:-
It is quite expensive to set-up a bio-fuel laboratory.
Bio-ethanol production demands strong technical knowledge for effective production and
also t avoid excess emissions.
Low production rate.
CHEMICALS REQUIRED:-
KMnO4
Distilled water
Saccharomyces cerevisiae (Yeast)
Sucrose
Urea

RAW MATERIALS REQUIRED:-

Fruit waste
PROCEDURE:-
YEAST ACTIVATION
FERMENTATION PROCESS
DISTILLATION
TESTING THE PROPERTIES OF BIO-ETHANOL
YEAST ACTIVATION:-
Take 200 ml of water in a beaker and heat the water around 35-40℃
Weigh 1gm of Urea ,40 gm of sucrose and 10 gm of Saccharomyces cerevisiae (Yeast)
simultaneously.
Add the above 3 components in the heated water and wait till 10-15 minutes then the
yeast will activates.
This mixture was taken as an inoculum for fermentation process.
FERMENTATION PROCESS:-
About 200g of Fruit wastes or rotten fruits such as apple, banana, papaya and grapes
were weighed separately and were taken in different beakers.
which was washed with 5% potassium permanganate solution and then rinsed with
distilled water.
pomegranate , sapota and papaya wastes were crushed separately in a mixer and
collected in beakers.
Take 1 liter conical flasks and the each fruit wastes are transfer to the flasks separately
Add the inoculum is also transferred to the conical flasks and the mixture is makeup to
1000 ml
Set the incubator temperature to 30°C. Wait till the set temperature is reached.
Keep the inoculum containing beaker inside, at the center of the incubator. close the
incubator door. Allow the temperature to equilibrate for 4-5 days .
During fermentation process, zymase an enzyme from yeast changes the simple
sugars into ethanol and carbon dioxide.

C₁₂H₂₂O₁₁+ H₂O ---invertase---> C₆H₁₂O₆ + C₆H₁₂O₆

C₆H₁₂O₆ ---zymase---> 2C₂H₅OH + 2CO₂ ↑

Take out the beaker from incubator after 72 Hrs. and continue for further processes.
DISTILLATION:-
Differential distillation is a procedure by which two liquids with different boiling
points can be separated.
After fermentation process is completed the sample was filtered with a thin cloth.
Fill the distillation flask with the filtered sample. The flask should be no more than two
thirds full because there needs to be sufficient clearance above the surface of the liquid.
Heat the distillation flask slowly until the liquid begins to boil. Vapors will begin to
rise through the neck of the distillation flask.
As the vapors pass through the condenser, they will condense and collected in the
receiving flask.
 Cool water in
Condenser

Distilling flask

Cool water out

Heater

Receiving
flask
Distillate
TESTING:-
Dichromate test for ethanol
Determination of Calorific value by Calorimeter
Determination of Concentration by UV-Spectrophotometer
Determination of Density by Hydrometer
DICHROMATE TEST:-
After distillation the product is placed in an air tighten flask, because the alcohol may
evaporate in the atmosphere.
For the test take a test tube then add potassium dichromate solution and acidified with
dilute sulphuric acid. A few drops of the alcohol is added to this solution.
The tube would be warmed in a hot water bath. In the case of a primary or
secondary alcohol.
First the solution is in orange color. When the solution is warmed the orange solution
turns green when the solution containes primary or secondary alcohol.
CALORIFIC VALUE TEST:-
 The crucible of the calorimeter is filled with the known mass of fuel, and then it is
ignited.
 This heats the surrounding water, and the initial and final temperatures are recorded
using a thermometer.
 The calorific value can be determined using the heat balance. Heat given by the fuel is
equal to the heat gained by the water.
 LHS: Mass of fuel × calorific value.
RHS: mass if water × specific heat ×change in temperature.

Equate LHS and RHS to get the calorific value.

CONCENTRATION TEST:-
 procedure for measuring ethanol concentration from yeast culture using K2Cr2O7
reagent (3.4g K2Cr2O7 +32.7ml H2SO4 ) in spectrophotometric method.
 The reaction mixture contains 0.1ml of culture supernatant and 2.5 ml of K2Cr2O7
reagent and heated for 10min at 60°C.
 Then, the solution is diluted with distilled water to make the volume 25ml and
measure the absorbance at 600nm to calculate the ethanol percentage from a standard
curve which was made by using 5%,10%,15%,20%,25% ethanol.
DENSITY TEST:-
 The hydrometer measures the weight of the liquid displaced by the volume of
the hydrometer.
 Specific Gravity is a dimensionless unit defined as the ratio of density of the
material to the density of water.
 If the density of the substance of interest and the reference substance are
known in the same units.
 Fill your hydrometer jar about ¾ with the liquid you wish to test. Insert the
hydrometer slowly.
 Now give it a spin with your thumb and index finger, this will dislodge any
bubbles that may have formed. Once the hydrometer comes to a rest, observe
the plane of the liquid surface.
 The point at which this line cuts the hydrometer scale is your reading.
 From the obtained density value calculate the sp.gravity.From the standard
graph between sp.gravity and concentration we can calculate the
concentration.
ex:- 100% ethanol has a specific gravity of .785 which is lighter than
water with a specific gravity of 1.0.
RESULTS & DISCUSSION:-
1)PERCENTAGE OF ALCOHOL:-
The flask contain 1000 ml of content and after distillation the various fruit wastes are
containing the following percentages of alcohol.
S.N FRUIT WASTE ETHANOL BY % OF ETHANOL
O VOLUME(ml)
1 POMEGRANATE 120 12
2 SAPOTA 150 15
3 PAPAYA 100 10
4 WATERMELON 80 8
 % OF ALCOHOL

16
14
12
10
8
6
4 % OF ALCOHOL
2
0
2)Dichromate test:-
In this test is conducted to confirm whether the obtained product is ethanol or not.
When the test is conducted we got the following results.

From the obtained results we confirmed that the obtained product is primary alcohol
nothing but ethanol.