CHE-005L: Organic Chemistry Laboratory

Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

ACTIVITY NO.7
FERMENTATION OF ALCOHOL

Learning Objectives
At the end of the activity, you will be able to:
• Describe the concept of fermentation in the preparation of ethanol.
• Explain the chemical reactions involved in alcoholic fermentation and its importance in the production of
alcoholic beverages.
• Demonstrate the principles of distillation as a method for separating and purifying ethanol produced
through fermentation.

Materials
• Fruit of your choice (e.g., banana, grapes, pineapple, mango, papaya, apple, or etc.), peeler (optional),
1-L glass bottle (e.g., Sprite or coke glass bottle), cotton, yeast, match, spatula, mortar and pestle,
graduated cylinder, test tube, test tube holder test tube rack, Erlenmeyer flask, stirring rod, watch glass,
dropper, distilling flask, universal/extension clamps, clamp holders, iron stands/ring stands, condenser,
wire gauze, thermometer, rubber tubings, funnel, filter paper, distilled water, potassium dihydrogen
orthophosphate or monopotassium phosphate (KH2PO4), limewater (1.6% Ca(OH)2), Iodine solution,
KOH T.S. (6.5% KOH solution) hot plate top-loading balance or analytical balance

Introduction

Fermentation is a metabolic process by which microorganisms, such as yeast or bacteria, convert

carbohydrates, such as sugars or starches, into various products, often including alcohol, acids, or gases. It is
an ancient technique used in food and beverage production, as well as in industrial applications such as biofuel
production and pharmaceutical manufacturing.

Alcoholic fermentation is a specific type of fermentation carried out by yeast, primarily the species
Saccharomyces cerevisiae, in the absence of oxygen (anaerobic conditions). It involves the conversion of
sugars, typically glucose or other fermentable carbohydrates, into ethanol (alcohol) and carbon dioxide. Alcoholic
fermentation is widely used in the production of alcoholic beverages such as beer, wine, and spirits, as well as
in the production of biofuels such as ethanol.

For these activities or objectives to be realized, fermentation of ethanol coming from a fruit source and
distillation of ethanol should be practiced in experimental laboratory.

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

A. Fermentation Process
Procedure:
1. Remove the peel or skin of the fruit of your choice (e.g., pineapple, banana, grape, or etc.).
2. Mince or dice the fruit using a spatula.
3. Extract the juice from the fruit by grinding the minced or diced fruit in the mortar and pestle.
4. Filter the extracted juice in a 1-L glass bottle (e.g., Sprite or Coke glass bottle) and discard the residue.
Collect at least 100mL of the fresh fruit juice (filtrate) in the glass bottle.
5. Put 5g of yeast into 25mL of distilled water and add this to the glass bottle.
6. In the glass bottle, add 1.4g of potassium dihydrogen orthophosphate or monopotassium phosphate
(KH2PO4) and 100mL of distilled water. (Note: if KH2PO4 is not available, only add 100mL of
distilled water).
7. Stopper the bottle with a cotton and let the mixture stand for one week at room temperature.
8. Write the chemical reaction involved in the OBSERVATION SHEET.
9. Take a picture of your output (Picture A) and make sure to also take a picture of your group (or
assigned member) with the output (Picture B). Place these pictures in the DOCUMENTATION
SHEET.

B. Test for Carbon Dioxide

Procedure:
1. Pour the gas from the glass bottle into a test tube containing 10mL of limewater (1.6% Ca(OH) 2
solution). Even though one cannot see the gas, it is denser than air and will pour into the test tube.
2. Stopper and shake the mixture in the test tube for 3-5 minutes.
3. Record any observations in the OBSERVATION SHEET.
4. Take a picture of your output (Picture A) and make sure to also take a picture of your group (or
assigned member) with the output (Picture B). Place these pictures in the DOCUMENTATION
SHEET.

C. Distillation Process
Procedure:
1. At this point, the fermented liquid is referred to as "wine" or "fruit wine” (e.g., pineapple wine, banana
wine, papaya wine, or etc.)". Decant the fermented liquid from the glass bottle into a distilling flask.
2. Assemble the distillation set-up and distill the mixture. Use an Erlenmeyer flask as your receiving
container.
3. Discard the first 1mL of distillate collected.
4. Record the temperature of the vapors in the OBSERVATION SHEET as soon as another 1 mL of
distillate has been collected.
5. Continue collection of the distillate. Collect at least 20-30mL of the distillate. Record the temperature
of the vapors at this point in the OBSERVATION SHEET.
6. Let the distillate cools down at room temperature.
7. Take a picture of your output (Picture A) and make sure to also take a picture of your group (or
assigned member) with the output (Picture B). Place these pictures in the DOCUMENTATION

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

SHEET.

D. Tests for Alcohol

Procedure:
1. Combustion Test:
a. Put 2mL of the distillate on a watch glass.
b. Use a match to ignite.
c. Record any observations in the OBSERVATION SHEET.
d. Take a picture of your output (Picture A) and make sure to also take a picture of your group (or
assigned member) with the output (Picture B). Place these pictures in the DOCUMENTATION
SHEET.
2. Iodoform Test:
a. Put 2mL of the distillate in the test tube and add 5mL of Iodine solution.
b. Add KOH T.S. (6.5% KOH solution) drop by drop until the solution become decolorized.
c. See the resulting mixture aside to an hour, and the note any formation of iodoform.
d. Record any observations in the OBSERVATION SHEET.
e. Take a picture of your output (Picture A) and make sure to also take a picture of your group (or
assigned member) with the output (Picture B). Place these pictures in the DOCUMENTATION
SHEET.

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

OBSERVATION SHEET

A. Fermentation Process.

1. Write/Illustrate the Chemical Reaction Involved.

B. Test for Carbon Dioxide.

1. What happens after introducing the gas from the fermented liquid into the lime water and
shaking it for 3-5 minutes in the test tube?

2. Write/illustrate the Chemical Reaction Involved.

C. Distillation Process

1. Temperature of vapor after collecting 1 mL of distillate ________

2. Temperature of vapor at the end of distillation ________

D. Tests for Alcohol

Complete the table below:
Name of the Test Theoretical Result Actual Result
Combustion Test

Iodoform Test

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

DOCUMENTATION SHEET

A. Fermentation Process. Place the pictures needed in their designated boxes.

PICTURE A PICTURE B

B. Test for Carbon Dioxide. Place the pictures needed in their designated boxes.

PICTURE A PICTURE B

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

C. Distillation Process. Place the pictures needed in their designated boxes.

PICTURE A PICTURE B

D. Tests for Alcohol. Place the pictures needed in their designated boxes.
1. Combustion Test

PICTURE A PICTURE B

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

2. Iodoform Test

PICTURE A PICTURE B

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

ACTIVITY QUESTIONS

1. What is Fermentation?

2. What is a yeast? What is its source?

3. What is an enzyme? Name 2 enzymes present in yeast and their functions.

4. Enumerate the different synonyms of alcohol.

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

5. What important by‐products are obtained in commercial fermentation processes?

6. Briefly discuss what does alcohol proof mean?

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

7. How are the following alcoholic beverages made?

a. Scotch

b. Rye

c. Bourbon

d. Rum

e. Vodka

This document is the property of PHINMA EDUCATION

 CHE-005L: Organic Chemistry Laboratory
Laboratory Activity Sheet #7

Name: _______________________________________________ Class number: ______

Section: ____________ Schedule: ________________________ Date: _____________

References:
• Stoker, H. S. (2013). General, Organic, and Biological Chemistry (6th ed.). Brooks/Cole, Cengage
Learning.
• Denniston, Katherine J. (2017). General, Organic, and Biochemistry (9th ed.) McGraw-Hill Education.
• Coyne, G.S. (1997). The Laboratory Companion: A Practical Guide to Materials, Equipment, and
Technique. John Wiley & Sons, Inc.
• Bettelheim, F.A., Landesberg, J.M. (2010). Laboratory Experiments for Introduction to General, Organic,
and Biochemistry (7th ed.). Brooks/Cole, Cengage Learning.
• Seager, S.L., Slabaugh, M.R. (2010). Safety Scale Laboratory Experiments for Chemistry for Today:
General, Organic, and Biochemistry (7th ed.). Brooks/Cole, Cengage Learning.

This document is the property of PHINMA EDUCATION