Wine Making

GROUP 1
3 A BIOCHEMISTRY

ABION, THEORELL JOSHUA J.

ALPAY, RIACARL F.
BERBOSO, MERIE JANE R.
CASUGA, JESSEL MAY D.

OBJECTIVES
To utilize Saccharomyces cerevisiae in wine
fermentation.
2. To employ different wine parameters for quality
assessment.
3. To compare our own product to a commercial
white wine in terms of the said parameters.
1.

INTRODUCTION
Winemaking a.k.a. Vinification is the production of wine, starting

from the selection of fruits (grapes, apple, etc.) and ending with
bottling of the finished wine.
Wine is produced by fermenting grape juice (which has particularly
high levels of sugar) using specialised yeast cells. The sugar is
converted into ethanol and carbon dioxide under anaerobic
conditions: (Alcoholic Fermentation)

C12H22O11 -----------> C6H12O6 ----------> CH3CH2OH

CO2
(invertase)
(Zymase)
Invertase and Zymase are enzymes produced by the yeast

INTRODUCTION
Malolactic fermentation (also known as malolactic conversion or MLF)

is a process in winemaking in which tart-tasting malic acid, naturally

present in grape must, is converted to softer-tasting lactic acid.
Often occurs shortly after the end of the primary fermentation but can
sometimes run concurrently with it. (Secondary fermentation)

Primary role in winemaking is to de-acidify the wine

Standard process for Red wine production and some white grape wine

(Chardonnay)

INTRODUCTION
Winemaking is not a simple step but a complex because there are many

insoluble particles suspended in the wine before bottling.

Fining substances added before bottling that can remove organic
materials and can improve aroma/flavor.
Aging
Flotation
Refrigeration
Pasteurization
Racking
Filtration
Preservatives- such as SO2

Bottling - final process in which the wine are traditionally sealed with a

cork.

METHODOLOGY
Remove the core of the apples
Place the apples in a zip lock bag
Leave in the freezer for three days
Extraction of Thaw the apples for one day
Fruit Juice Extract the juice by squeezing them in a cheesecloth

Fermentation

Pasteurization

Transfer the juice in a clean Erlenmeyer flask

Inoculate the yeast (Saccharomyces cerevisiae)
Cover the flask and leave it in a dark place to ferment

Decant the fermented juice through a cheeseloth

Prepare Bentonite by boiling it with water
Add the Bentonite in the fermented juice
Leave the juice for 3 days or until the juice is cleared up

METHODOLOGY
Filter the mixture into a clean flask or beaker
Warm at 85C for 1 minute
Pasteurization Cool at 50C for 3 days

Bottling &
Testing

Transfer the pasteurized wine in a clean and pre-sterilized

bottle
Test for the basic wine parameters
Label the wine with its basic parameters

RESULTS AND DISCUSSION

AMOUNT OF REDUCING SUGAR
Abbe Refractometer was used
Commercial wine had a value of 1.3479. therefore,

Novellino contains 10% amount of reducing sugar

Produced wine had a value of 1.3348. therefore,
apple wine contains 5% amount of reducing sugar

RESULTS AND DISCUSSION

AMOUNT OF REDUCING SUGAR

RESULTS AND DISCUSSION

RELATIVE DENSITY
The specific gravity of a wine is the density of a wine in

relation to the amount of dissolved sugar in it.

The MORE SUGAR the HIGHER the Specific gravity (less
alcohol) and LESSER SUGAR means LOWER the Specific
gravity (more alcohol)
SG of 0.900 to 1.000 Dry
SG of 1.000 to 1.010 Medium Sweet
SG of 1.010 to 1.025 Sweet
SG of 1.025 and up
Very Sweet

RESULTS AND DISCUSSION

RELATIVE DENSITY
Commercial white wine: 0.
9198g/mL

Produced wine: 1.00965g/mL

The commercial white wine The produced white wine is

(Novellino) is therefore a DRY concluded to be a MEDIUM
wine; having more sugar
SWEET wine, therefore
present with less alcohol
having HIGHER SUGAR
concentration on it.
content and a lower alcohol
concentration.

RESULTS AND DISCUSSION

pH Value

Basic Principle

pH meter determines how acidic or basic a liquid is and the

concentration of hydrogen ions.

Acids have a pH less than 7.0 while bases have a pH higher than 7.0.
Plain water measures 7.0. Most wines fall between 3.1 and 3.6.
If the range falls below 3.1, the wine is highly acidic, tastes too tart
If the range is above 3.6, the wine will be oxidized to and microbial
growth is possible

pH is the backbone of Wine Alison Crowe (Winemaker Magazine)

RESULTS AND DISCUSSION

pH Value
Malic acid

White wine grapes or light coloured fruit wines such as

apple wines or ciders should have a pH close to 3.2 (3.1 to

3.3).
Wines can be de-acidified by adding calcium carbonate or
potassium carbonate. Addition of an acid blend containing
malic, citric, and tartaric acids to acidify
pH is the backbone of Wine Alison Crowe (Winemaker Magazine)

RESULTS AND DISCUSSION

pH Value
Theoretical pH values (white wine)
pH 3.1-3.3
Experimental (Novellino & Mansuetus)
pH 4.0

pH is the backbone of Wine Alison Crowe (Winemaker Magazine)

RESULTS AND DISCUSSION

Percent Alcohol
Ebulliometer
-designed to accurately measure the
boiling point (temperature) of water or a
water solution, by measuring the
temperature of the vapour generated
away from the heat source.
-Boiling point of dH2O: 99.8C
-Boiling point of wine sample: 97.1C
= 99.8C 97.1C = 2.70
% alcohol = 2.90

RESULTS AND DISCUSSION

Percent Alcohol

RESULTS AND DISCUSSION

Percent Alcohol

RESULTS AND DISCUSSION

Percent Alcohol

RESULTS AND DISCUSSION

Free and Total SO2
Ripper Method
- uses an iodine standard to titrate the SO2 in a sample. Free SO2 is determined
directly while total SO2 can be ascertained by treating the sample with sodium
hydroxide (before titrating) to release bound SO2.
- the free and total SO2 analysis used in the Ripper test is based on the reaction:
SO2 + I2 + 2H2O H2SO4 + 2 HI
- a starch indicator is added to the wine sample and it is acidified with H2SO4. The
sample is then rapidly titrated with an iodine solution. The completion of the
reaction is noted when excess iodine is complexed. This is determined by a blueblack color end point in the presence of a previously added starch indicator.

RESULTS AND DISCUSSION

Free and Total SO2
-only a proportion of the SO2 added to a wine will be effective as an anti-oxidant.
The rest will combine with other elements in the wine and cease to be useful. The
part lost into the wine is said to be bound, the active part to be free.
Red wines do not need any added sulphur dioxide because they naturally contain
anti-oxidants, acquired from their skins and stems during fermentation.
White wines and ross do not contain natural anti-oxidants because they are
not left in contact with their skins after crushing. For this reason they are more
prone to oxidation and tend to be given larger doses of sulphur dioxide.

Sweet wines get the biggest doses because sugar combines with and binds a high
proportion of any SO2 added. To get the same level of free sulphur dioxide, the
total concentration has to be higher than for dry wines.

RESULTS AND DISCUSSION

Free and Total SO2

Before

After

RESULTS AND DISCUSSION

Free and Total SO2

Free SO2

ppm SO2 = vol iodine solution x coefficient

= 1.32 x 12.8 = 16.896 ppm = 16.876 mg/L (Novellino)
= 0.096 x 25.6 = 2.458 ppm = 2.455 mg/L (Mansuetus)
Total SO2
ppm SO2 = vol iodine solution x coefficient
= 5.11 x 32 = 163.52 ppm = 163.33 mg/L (Novellino)
= 0.8 x 64 = 51.2 ppm = 51.142 mg/L (Mansuetus)

RESULTS AND DISCUSSION

Free and Total SO2

RESULTS AND DISCUSSION

Titratable Acidity
Total acid in a wine is a factor in proper aging and in the
sensory impression of a wine. When acid is very important
as it contributes to the color, stability and sensory properties
of wines. Tartaric acid and malic acid are the two major
acids in wines and account for about 90% of a wines total
titratable acidity.

RESULTS AND DISCUSSION

Titratable Acidity
Titratable Acidity
The Titratable acidity of a solution is an

approximation of the solution's total acidity.

The titratable acidity of a solution is measured by
reacting the acids present with a base such as sodium
hydroxide(NaOH) to a chosen end point, close to
neutrality, as indicated by an acid sensitive colour
indicator.
Titratable acidity, in regards to wine production, is
given in units of g/L as per tartaric acid equivalents.

RESULTS AND DISCUSSION

Titratable Acidity
Tartaric Acid Equivalents
Titratable acidity of a solution, is a measure of all the various
acids present in solution.
The calculations performed to determine the grape juice or wine's
acidity, following the titration procedure, assumes, for
convenience, that all acids exist as tartaric acid.
Tartaric acid is chosen to represent the other acids present, as
this is usually the main acid in grape juice and wine.
The units of titratable acidity then is given in g/L as tartaric acid
or tartaric acid equivalents.
In countries such as France, acidity concentrations are give as g/L
of sulphuric acid (H2SO4) equivalents. A much smaller figure.

RESULTS AND DISCUSSION

Titratable Acidity
Standardization:

Titratable Acidity:

RESULTS AND DISCUSSION

Volatile Acidity
Volatile acid at high levels is often regarded as an indication of spoilage.
Volatile acidity, most of which is acetic acid, can be a problem in wine.
Pronounced volatile acidity to the nose usually indicates that the
bacterium Acetobacter has contaminated the wine, changing some
ethanol to acetic acid by an oxidation process. As wine ages and
undergoes slow oxidation, some ethanol is oxidized to acetic acid or
acetaldehyde affecting the sensory impression of the wine. There is a
legal limit for volatile acidity, though most wines are unfavourably
judged before the legal limit is reached. Thus, at bottling, or if
Acetobacter contamination is suspected, a volatile acid test is done. The
easiest and most accurate methods are chromatographic or enzymatic,
but require extremely expensive equipment. In the measurement for
volatile acidity a steam distillation method is used.

RESULTS AND DISCUSSION

Volatile Acidity
Steam distillation

SET-UP
c) End point with lightest

yellow color

RESULTS AND DISCUSSION

Volatile Acidity
Volatile acidity refers to the steam distillable acids
present in wine, primarily acetic acid but also lactic,
formic, butyric, and propionic acids. Commonly, these
acids are measured by Cash Still, though now they can be
measured by gas chromatography, HPLC or enzymatic
methods. The average level of acetic acid in a new dry
table wine is less than 400 mg/L, though levels may
range from undetectable up to 3g/L.
U.S. legal limits of Volatile Acidity:
Red Table Wine 1.2 g/L
White Table Wine 1.1 g/L

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