Laboratory Experiments in Chemistry for Engineers

EXPERIMENT 4

Fuels: Preparation of Bioethanol

OBJECTIVES
 To produce bioethanol by fermentation process.
 To determine the flammability property of bioethanol.

DISCUSSION
Bioethanol production involves a biological process of fermentation
wherein sugars are converted to ethanol and carbon dioxide by the action
of yeast. Microorganisms such as yeast affect the amount of ethanol yield
as well as other factors like temperature, sugar concentration, pH and
fermentation time.

Sucrose (C12H22O11), a disaccharide is hydrolyzed into monosaccharides

fructose and glucose by the enzyme invertase present in yeast. Through a
series of reactions, the enzyme then converts fructose and glucose into
ethanol and carbon dioxide. The phosphates in the fermentation solution
increase yeast growth and promote fermentation process.

High concentration of alcohol solutions are obtained by fractionally

distilling the aqueous alcohol solution produced by fermentation.
Although, it will not yield 100% pure ethanol because of the formation of
azeotropic mixture of 95% ethanol and 5% water.

Set-up for the fermentation of sucrose

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Laboratory Experiments in Chemistry for Engineers

MATERIALS

 Big test tube  Test tube

 pipet  Hot plate
 Evaporating dish  glass tubing
 stirring rod  funnel
 (3) filter paper  beaker 250 mL
 thermometer  rubber stopper

REAGENTS

Substance Description
 sucrose solid
 baker’s yeast solid
 disodium hydrogen solid
phosphate
 calcium hydroxide solution
 ethanol liquid
 distilled water for dilutions

SAFETY PRECAUTIONS
Avoid skin contact and inhalation of vapors.
Wash hands in case of skin contact.

PROCEDURE
A. Fermentation of Sucrose

1. In a top-loading balance, weigh out 1.30g of sucrose. Place it on a big

test tube and add 3.75 mL of water.

2. Add 0.1 g of baker’s yeast.

3. Stir until the sugar dissolves and no yeast granules remain.

4. Then add 0.1g of disodium hydrogen phosphate and 1.25 mL of water.

5. Warm the mixture to about 45oC.

6. Shake this mixture to ensure complete mixing and then fit the neck of
the test tube with a glass tubing connected to another test tube.

7. The glass tubing should be beneath the surface of about 2 mL of

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Laboratory Experiments in Chemistry for Engineers

saturated aqueous solution of calcium hydroxide (limewater) in a reaction

tube. The tube in the limewater will act as a seal to prevent air and
unwanted enzymes from entering the flask but will allow gas to escape.

8. Place the set-up in a warm spot (the optimal temperature for the
reaction is 35oC) for a week, at which time the evolution of carbon dioxide
will have stopped.

9. After the completion of fermentation, filter the mixture. Wash out the
test tube with 2 mL of water and rinse the filter paper.

10. Collect the filtrate.

11. Under the teacher’s supervision, pour the filtrate into an evaporating
dish and light it with a match. Compare it to the burning of reagent grade
ethanol.

DISPOSAL AND CLEAN-UP:

All aqueous solutions can be discarded into the sink. Flush with generous
amounts of tap water. Clean your lab table after the experiment.

REFERENCES:

 Mohrig, Jerry R. Experimental Organic Chemistry, A Balanced

Approach: Macroscale and Microscale, New York: W.H.
Freeman and Company, 1998.
 Williamson, Kenneth L. Macroscale and Microscale Organic
Experiments, 2nd ed. USA: D.C. Heath and Company, 1994.

NAME: ____________________________

Experiment 4  Fuels: Preparation of Bioethanol 8

Laboratory Experiments in Chemistry for Engineers

GROUP: _________RATING:____________
PRELAB ACTIVITY
FOR EXPERIMENT DATE SUBMITTED: ___________________

4
________________________________________________________________________

Fuels: Preparation of Bioethanol

1. Define fermentation and show the chemical reaction of the conversion of sucrose to
ethanol and carbon dioxide.

Fermentation is a metabolic process that converts carbohydrates, such as sugars and

starches, into alcohol or organic acids using microorganisms like yeast or bacteria under
anaerobic conditions. This process is utilized in various industries, including food and
beverage production, biofuel manufacturing, and pharmaceuticals.

Chemical Reaction: Conversion of Sucrose to Ethanol and Carbon Dioxide

The chemical reaction for the conversion of sucrose (C12H22O11) to ethanol (C2H5OH)
and carbon dioxide (CO2) through fermentation by yeast can be represented as follows:
C12H22O11 → 4C2H5OH + 4CO2

This equation illustrates the breakdown of sucrose into four molecules of ethanol and
four molecules of carbon dioxide. The process involves the enzymatic action of yeast on
the sucrose molecule, resulting in the production of ethanol and carbon dioxide as
byproducts.

In this reaction, the yeast utilizes the sucrose as a source of energy, breaking it down into
simpler compounds through a series of biochemical reactions. The primary steps involve
the conversion of sucrose into glucose and fructose, followed by glycolysis, where these
simple sugars are further metabolized to produce ethanol and carbon dioxide.

2. What is the difference between simple distillation and fractional distillation?

Simple distillation and fractional distillation are two common techniques used to separate
mixtures based on the differences in boiling points of the components. While both
methods involve the process of vaporization and condensation, they have distinct
differences in their application and efficiency.
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Laboratory Experiments in Chemistry for Engineers

In summary, while both simple and fractional distillation are methods for separating
mixtures based on boiling point differences, fractional distillation offers higher efficiency
and is better suited for mixtures with closely related boiling points.

3. Show the set-up of A) simple distillation and B) fractional distillation and label the
parts.
Set-up:
The simple distillation set-up consists of a round-bottom flask, a distillation head, a
thermometer adapter, a Liebig condenser, and a receiving flask.
Labeling the Parts:
Round-bottom flask: Holds the liquid mixture to be distilled.
Distillation head: Connects the round-bottom flask to the condenser.
Thermometer adapter: Allows the insertion of a thermometer to monitor the temperature
of the vapor.
Liebig condenser: Cools the vapor back into liquid form.
Receiving flask: Collects the distilled liquid.
B) Fractional Distillation:
Set-up:
The fractional distillation set-up includes a fractionating column in addition to the
components of simple distillation.
Labeling the Parts:
Fractionating column: Contains multiple plates or packing material to provide more
surface area for repeated vaporization and condensation, allowing for better separation of
components based on their boiling points.

Experiment 4  Fuels: Preparation of Bioethanol 10

Laboratory Experiments in Chemistry for Engineers

NAME: ____________________________

GROUP: _________RATING:____________

LAB REPORT FOR DATE PERFORMED: __________________

EXPERIMENT 4 DATE SUBMITTED: ___________________

________________________________________________________________________

Fuels: Preparation of Bioethanol

RESULTS AND OBSERVATIONS

Observation
1. solution in the test tube containing When calcium hydroxide is added to a
Ca(OH)2 solution, the pH rises, a white precipitate
forms, and the solution may become clearer
as a result of neutralisation or the formation
of compounds that are only sporadically
soluble.
2. filtrate when lighted with a match When a filter is lit with a match,
observations include the possibility of
residue signifying non-combustible
materials, the potential for ignition if the
filtrate is flammable, and the colour or
characteristics of the flame that reveal
components present.
3. reagent grade ethanol when lighted with When Reagent Grade Ethanol is lit with a
a match match, one should anticipate a clear, blue
flame, little to no soot or residue, and a
steady, constant flame because of the high
ethanol purity.

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Laboratory Experiments in Chemistry for Engineers

LAB REPORT FOR EXPERIMENT 4 (continued) NAME:

POST-LAB QUESTIONS

1. What are yeasts? Explain the role of yeast in the fermentation process.
A microorganism called Saccharomyces cerevisiae is used in the fermentation process to
make alcohol. Because yeast has enzymes like zymase that can metabolise carbohydrate
molecules without oxygen, it is used in fermentation. Ethanol and carbon dioxide
molecules are produced as a result.

2. Write the equation for the formation of the precipitate formed in the solution
containing calcium hydroxide.
Ca(OH)₂ (aq) + Na₂SO₄ (aq) → CaSO₄ (s) + 2NaOH (aq) In this equation, (aq)
represents an aqueous solution, and (s) represents a solid precipitate.

3. What other raw materials can be used for the production of bioethanol?
Six primary categories comprise the raw materials containing lignocellulose needed to
produce bioethanol: crop residues (such as cane and sweet sorghum bagasse, corn stover,
various straw types, rice hulls, olive stones, and pulp), softwoods (such as pine and
spruce), hardwoods (such as aspen and poplar), and cellulose wastes.

SUMMARY AND CONCLUSIONS

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