GROUP 4

Abo

Pepito

Dorol

Salvacion

Garcia
EXPERIMENT 1: WINEMAKING
I.

Introduction

OBJECTIVES:
Upon completion of this experiment, the student should be able to:
1.
2.
3.
4.

Simulate the process of fermentation in wine-making.

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

Wine over the course of time has been used in many aspects in life such as in
the social and religious context in society. It has been used as a beverages for
occasions, a representation of the blood of Christ during a catholic mass and now, it
is said that drinking wine is equivalent to an hour at the gym. Wine is produced by
fermenting a fruit juice, for example a grape juice, by an active yeast. The yeast
which is a member of the fungi family converts the sugar into alcohol (ethanol) and
carbon dioxide together with other by-products.

The
sugars,
phenolic, and organic acids give the juice its flavor which comes from the
fermentation process by the yeast. These give the flavor of the wine because some
oxygen present is converted into an acid and eventually form to be an ester. The
ester will be the flavor compounds that is present in the wine. The vitamins,
minerals and nitrogenous compounds are essential to the yeast growth and

fermentation. The nitrogenous compounds are important building blocks for yeast
during fermentation while the minerals like the magnesium and potassium is used
as a co-factor for the production of certain enzymes required for the process.
If the juice that will be used is a grape juice which has 79% water, 20%
carbohydrates, 1% organic acids and trace amounts of other compounds the wine
will have a similar composition but lower sugar levels, 8% to 13% alcohol and
greater range of minor components.
Commercial wine manufacture has five basic steps. Step 1 is harvesting where
the grapes are harvested when its compounds are optimized for the style of wine
desired. Step 2 is crushing and de-stemming. Step 3 is pressing where the juice is
extracted. Step 4 is fermentation and is inoculated with active yeast. The last step,
step 6, is purification where unwanted solids, salts, and microorganisms are
removed.

II.

Methodology

First, the working area is sanitized as well as the apparatus and other materials
to be used such as the corks and stoppers. In the sterilization or sanitization phase,
the percentage of the laundry bleach or sodium hypochlorite is checked so that an
adequate level can be maintained. A 500mL of sodium hypochlorite is to be
prepared for sanitization but a disinfectant alcohol was used instead for the work
area and boiled water for the glassware and cork.
For determining the titratable acidity, TA, the orange juices (the juice assigned
to the group) pH was recorded. The burette was then filled with 0.1 sodium
hydroxide solution and the initial volume was recorded. Next, in an Erlenmeyer flask
hot distilled water of 50 mL was placed and 5 mL of juice to be tested as well. Five
drops of phenolphthalein indicator was next. Then the water-juice solution was
titrated with the sodium hydroxide by slowly adding the titrant to the Erlenmeyer

flask and stirred well. The procedure was stopped when there was evident color
change, it turned a darker color and was no longer pure orange but a color that is
nearly dark red. The used volume was recorded after the titration process. This was
done two times and the titratable acidity was computed.
The next procedure was to acidify the juice where 0.60L or 600mL of the orange
juice was boiled for five to ten minutes. It was cooled by placing it on an ice bath.
Both the temperature of the juice when boiled and cooled was recorded. Due to the
pH pen used, adding of the malic acid or citric acid to acidify the juice was no longer
done. The third procedure was to raise the sweetness of the juice by computation of
the sugar needed to add. After the sugar was added, the solution was stirred well to
dissolve the sugar.
Addition of 0.93 mL of 10% sodium metabisulphite was done next into the
sweetened juice and stirred well. The pH of the starting solution was recorded and
the temperature. The solution was put on an ice bath and left to reach the 25C
temperature. After, addition of 0.30 mL of pectic enzyme was done and stirred well.
On a separated beaker, 2 grams of active dry yeast was weighed. The yeast was
hydrated with water and stirred until all the solids was dissolved. The solution was
left to settle for 3 minutes or until a foam was formed. The pH of that solution was
recorded. The dissolved active dry yeast was poured to the sweetened, sulfated
juice carefully and was transferred onto the sanitized wine bottle and was closed by
the cork with the Tygon or surgical tubing. The prepared wine was then fermented
for three to four weeks or until gas formation has stopped in an air-conditioned
room.

III.

Results and Discussion

Table 1.1 Determination of titratable acidity

Type of juice used
Initial pH of juice used
Initial volume of 0.1 N NaOH, mL
Final volume of 0.1 N NaOH, mL
Used volume of 0.1 N NaOH, mL
Grams per liter of tartaric acid

Orange Juice
3.2
0
7
7
1.05

Table 1.2 acidifying the juice

Boiling temperature of
54
juice, C
Temperature after
28.9
cooling, C
pH reading
3.1
Table 1.3. Raising the sweetness of the Juice
Total sugars of the juice (see label

12

8
15.1
7.1
1.065

panel)
Total weight of juice, grams
Total sugar added, grams
Final % of sugar in juice solution
Table 1.4. Sulfating and adding pectic
pH of starting solution
Initial Temperature of starting
solution, 0C
Final Temperature of starting
solution, 0C

6.573
142.09
23%
enzyme to the Juice
3.2
31
24

Table 1.5. Adding yeast to the Sweetened Juice

pH of yeast starter solution
Variations in determining the titratable acidity of the fermenting orange juice
during the fermentation were shown in table 1.1. Table 1.1 shows what kind of juice
was used and the volumes needed for determining the TA, more importantly the
used up volume of the NaOH. Titratable acidity plays an important role as a
parameter to determine the fruit maturity and sour taste of citrus fruits. One of the
most important factors to determine how the fruit will taste is by knowing its
maturity. Mature fruits normally have a high sugar to acid ratio compared to an
immature fruit. The initial pH is generally 3.2. The point of titratable acidity is at 7
mL in trial 1 and 7.1 in trial 2 which is almost the same. During the acidifying of
juice, the boiling temperature yields to 54 C and the temperature after cooling is
28.9 C. Generally, there was a slight decrease in pH from 3.2 to 3.1. A drop in pH
corresponds to an increase in titratable acidity, where acidity plays a vital role in
determining the quality of wine. A low pH enhances production of fruity esters
during yeast fermentation and can provide a fresher taste. The lack of acidity
means a poor fermentation. In the case of having lower acidity, study shows that it
can lead to the growth of spoilage organism. The titratable acidity is determined by
multiplying the used volume of the NaOH by the factor 0.15 which resulted to 1.05
for trial 1 and 1.065 for trial 2. The possible sources of error is pH recorded due to
the pH pen used and the multiplying factor for the orange juice since the 0.15 is for
a wine that is made from grape juice.
In raising the sweetness of the juice, a large amount of sugar was added (of
about 142.09 g). The reason for it is that the juice that we used for fermenting has
no added sugar content. ). The reason for it is although the orange juice has natural
sugar in them, the produced wine will have low alcohol content and the wine will not
keep for long. The sugar will also give the desired sweetness. Sugar in wine plays a
vital role, where one of these is in fermentation process where the yeasts
metabolize sugars for energy that yields to a byproduct as alcohol. In this case, to
find how much sugar will be added to the original total sugar content, the mass of

juice (determined by mass balance), volume of juice, density and its grams per 100
grams of the original juice was needed.
After a week, the wine has its primary fermentation. There is a high level of
sugar and oxygen during primary fermentation. As fermentation progresses, there a
drop of sugar levels and an alcohol (ethanol) and carbon dioxide started to build up.
The fermentation is said to slow down thats why primary fermentation is faster
than secondary fermentation.
The chemicals that is added during the experiment has importance in improving the
wine quality. Yeast is the most important in wine making because it distinguishes
wine from the orange juice. Yeast converts sugar of juice into alcohol and carbon
dioxide though the presence of fermentation. It is the reason why fermentation is
possible. Sodium metabisulphates role is to destroy wild molds and bacteria that is
present in the juice. Addition of pectic enzyme breaks down the pectin to prevent
pectin haze from forming so that the wine is easier to clear.

IV.

Summary and Conclusion

Overall, this experiment taught us that fermentation is a very useful process.

Using orange juice as our main ingredient in this wine made our wine acidic, but this
showed a good effect because acidity plays a vital role in determining the quality of
a wine. A low pH enhances production of fruity esters during yeast fermentation and
can provide a fresher taste. Because we used a concentrated pure orange juice, a
large amount of sugar was added into the wine. Sugar is important in this
fermentation process because it is the ingredient that is converted into an alcohol.
Another important ingredient in this experiment is the yeast. The yeast played a
very important role in this experiment because it is the one which converts sugar
into alcohol which is used to convert fruit juices into wine.
In conclusion, it is a successful experiment because our orange wine started
to ferment in just two weeks. The solution is not that clear and translucent but
theres a formation of bubbles at the top of the solution, which shows that CO 2 was
formed, which is a good indication of fermentation.
V.

References
Alton, R. (1999). Water Water Everywhere. Retrieved from
http://www.rpi.edu/dept/chem-eng/Biotech-Environ/beer/water2.htm
Keller, J. B. (2010). Acidity in wines. Retrieved from
http://winemaking.jackkeller.net/acid.asp
LaVilla, J. (1998). How does wine get its flavor?. Retrieved from
http://www.foodwine.com/food/egg/egg1197/wineflav.html

Rotter, B. (n.d.). How long do primary and secondary fermentations last?.

Retrieved from http://winemakersacademy.com/long-primary-secondaryfermentations/
The Influence of pH. (n.d.). Retrieved from
http://www.brsquared.org/wine/Articles/pH.htm