CHM150L Experiment 1

Winemaking

Objectives

Upon completion of this experiment, the student should be able to:

1. Simulate the process of fermentation in wine-making

2. Understand the principles behind catabolic fermentation
3. Utilize titration techniques as verification procedure in wine-making
4. Demonstrate the relevance of sanitation in the fermentation process.

LIST OF MATERIALS

100% Grapejuice hot plate

Glass bottles (1 liter capacity) Sanitary suction bulb
Cork or rubber stopper with hole Glass pipette
Tygon tubing or Surgical tubing Top loading balance
Active Dry yeast (Saccharomyces cerevisiae) Stirring rod
Laundry bleach 400 mL beaker
Sugar 250 mL beaker
4” long glass tubing Burettte Clamp
Test tube Distilled water
Masking tape pH meter
Distilled water burette clamp
Pectic enzymes (pectinase) burette
Iron stand Iron ring

LIST OF CHEMICALS

Sodium hydroxide Phenolphthalein indicator

Sodium metabisulphite Sodium hypochlorite (dilute bleach)
Citric acid or Malic acid or Cream of tartar

INTRODUCTION

Wine in the simplest terms is the product of the fermentation of fruit juice by yeast. During fermentation,
yeast converts sugar into roughly equal amounts of alcohol (ethanol) and carbon dioxide (CO2) along with
numerous other by-products.

Wine has importance in many social and religious contexts in society today, as it has for thousands of
years. The basic fermentation process whereby alcohol is produced from the sugar in grapes is very
simple, but its chemistry is still not completely understood. As this knowledge increases, winemakers are
being helped to improve the quality of their wine.

Grape juice consists of 79% water and 20% carbohydrates, 1% organic acids and trace amounts of organic
acids, phenolics, vitamins, minerals and nitrogenous compounds. The sugars, organic acids and phenolics
give the juice its flavor, while the vitamins, minerals and nitrogenous compounds are, in many cases,
essential to yeast growth and fermentation. Wine has a similar composition, but has much lower levels
of sugar (none in dry wines), 8-13% alcohol and a greater range of minor components. Commercial wine
manufacture consists of five basic steps.

Step 1 – Harvesting

This is the most critical stage of the process. The grapes must be harvested when the sugar, acid, phenol
and aroma compounds are optimized for the style of wine desired.

Step 2 – Crushing and destemming

The grapes are removed from the stems and gently crushed to break the skins. Sulfur dioxide is added to
the grapes at this stage to prevent oxidation and inhibit microbial activity. Enzymes may also be added to
break down the cell walls and aid the release of juice.

Step 3 – Pressing

The juice extraction process depends on the type of wines to be used, but always involves squeezing the
berries. After pressing, the juice is allowed to stand to separate the solids. If necessary, the juice may be
clarified by filtration or centrifugation.

Step 4 – Fermentation

The juice is inoculated with live yeast, which then carries out the fermentation reaction:

C6H12O6 → 2C2H5OH + 2 CO2

This reaction occurs through many intermediary biochemical steps. The process is carried out under a
blanket of carbon dioxide as in the presence of oxygen, the phenols are oxidized and the sugar and ethanol
are converted to carbon dioxide and water.

Step 5 – Purification

Unwanted solids, salts and microorganisms are removed through a variety of physical processes, then the
wine is bottled and sold.

The laboratory is involved throughout the process, and particularly at the harvesting, fermentation and
purification steps, which need to be monitored closely.

Winemaking could present a significant environmental problem as a large volume of waste with a high
BOD (biological oxygen demand) is produced, but this is usually composted or otherwise disposed of on
site, so the environmental hazard is minimized.
PROCEDURE

Note: Observe sanitation and sterilization of all apparatus and work area before conducting the
experiment. Observe good laboratory practices. Boil corks and stoppers in water to sanitize them.

Sterilization:

1. Check the percentage of sodium hypochlorite, the laundry bleach.

2. Prepare a 500 mL 10% solution of sodium hypochlorite by adding distilled water. Use this solution
to sanitize glasswares and other utensils.
3. Set aside hypochlorite solution in covered bottles.

Determining the Titratable Acidity (TA)

1. Get the pH of the juice. Record your reading.

2. Fill the burette with 0.1N sodium hydroxide solution and allow a small amount to run through.
Record the sodium hydroxide (NaOH) level in the burette.
3. Place bout 50 milliliters of hot distilled water in the beaker.
4. Add 5 mL of juice to be tested to the hot water using the pipette.
5. Add 5 drops of the phenolphthalein solution to the water-juice mixture.
6. Slowly add the sodium hydroxide from the burette to the mixture, stirring with the glass rod. Stop
the sodium hydroxide addition when the mixture turns pink.
7. Record the sodium hydroxide level in the burette, and calculate the number of milliliters used.
Multiply the volume of NaOH used by 0.15. The product is the percentage of titratable acidity of
the sample expressed as grams tartaric acid per L (g/L).

Acidifying the Juice

1. Boil 0.60L of grapejuice for 5-10 minutes. Let it cool down by stirring and complete the cooling
down by placing it in an ice bath.
2. Based on the initial pH and titratable acids (%) obtained, acidify the juice with citric or malic acid
or cream of tartar.
3. Add 0.26 gram of the acid into the cooled juice. Check the pH. Add more acid in 0.3 gram
increments until the pH reaches 3.4. Record the weight of total acid used.

Raising the sweetness of the Juice

1. Looking at the total sugars in the nutrient information label of the grapejuice, calculate the total
sugar needed to be added to raise the total sugar to 23%.
2. Add the calculated sugar into the juice. Mix well until all sugar has dissolved.

Sulfating and adding pectic enzyme to the Juice

1. Add 0.93 mL of 10% sodium metabisulphite solution into the sweetened juice.
2. Stir properly to disperse the chemical into the solution.
3. Get the pH of this starting solution.
4. Get the temperature.
5. Put the solution in an ice bath. Leave the solution until it reaches 25 0C.
6. Add 0.30 mL of the pectic enzyme into the solution. Stir well.
Adding yeast to the Sweetened Juice

1. Prepare the setup below.

2. Weigh 2.0 grams of Active Dry Yeast (ADY) in a beaker.
3. Hydrate the yeast by adding 30 mL of distilled water. Stir until all the solids are dissolved.
4. Leave the solution for about 3 minutes or until a foam is formed on the surface of the solution.
Get the pH of the yeast starter solution.
5. Pour the dissolved ADY into the prepared sweetened, sulfated juice. Make sure that you do the
transferring carefully. Make sure that the transfer is done with minimal air exposure.
6. Put the cork or rubber stopper. Place the setup in an airconditioned room. Monitor daily. Make
sure that the water in the exhaust tube is available at any given time.

Fermenting the Juice

1. Ferment the juice for 3-4 weeks or until gas formation has stopped. Note the presence of bubbles
on the surface of the solution.
2. Make sure that the jar is placed at a temperature range of 19-25 0C (aircon room).

“Racking” the Wine

1. Prepare and sanitize a glass bottle by using the 10% sodium hypochlorite bleach prepared from
previous procedure. Dry the bottle.
2. Open the primary jar and quickly place a Tygon or Surgical tubing with the end placed on the
supernatant liquid above the sediments and the other end on the secondary jar from Procedure
1. (See figure below.)

The wine produced from the fermentation is then aged for 1 year or longer. If the wine produced is semi-
clear, proper clarifying agents such as gelatin, charcoal, milk, bentonite or egg whites may be used. It will
be left to stand for another week and proper filtration methods shall be used to obtain a clear wine.

Furthermore, in order to develop a more characteristic wine, these liquids are stored in a cool, well
ventilated dark place.
 CHM017L. Experiment 1. Winemaking

Name: ______________________________________ Date Performed: __________________________

Course: _____________________________________ Date Submitted: ___________________________
Section: _____________________________________ Instructor: _______________________________

REPORT SHEET
Table 1.1. Determining Titratable Acidity

Type of Juice used

initial pH of Juice used
Initial volume of 0.1N NaOH, mL
Final volume of 0.1N NaOH, mL
Total volume of NaOH used
Grams per liter of tartaric acid

Table 1.2. Acidifying the Juice

Boiling temperature of juice, 0C

Temperature after cooling, 0C
pH reading
First addition of acid
Second addition of acid
Third addition of acid
Fourth addition of acid
Total weight of acid added, grams
Final pH of acidified juice

Table 1.3. Raising the sweetness of the Juice

Total sugars of the juice (see label panel)

Total weight of juice, grams
Total sugar added, grams
Final % of sugar in juice solution

Table 1.4. Sulfating and adding pectic enzyme to the Juice

pH of starting solution
Initial Temperature of starting solution, 0C
Final Temperature of starting solution, 0C
Table 1.5. Adding yeast to the Sweetened Juice

pH of yeast starter solution

Table 1.6. Fermenting the Juice

Day Number Remarks

Day 1
Day 3
Day 5
Day 7
Day 9
Day 11
Day 13
Day 15
Day 17
Day 18
Day 20
Day 22
Day 24
Day 26
Day 28

Racking the Wine

1. Describe the sensory properties (color, odor and taste) of the wine collected.