The Analysis of Three Types of Wine

Aims
My investigation aims to analyse a selection of the many
components of wine.

• What is the ethanol content (Alcohol by Volume in

percent) of each sample of wine?
• Is the ethanol content figure given on the bottle
correct?
This will be measured in two ways. The first method is the
least accurate of the two and looks at measuring the %
ethanol content of a sample of wine which still contains
volatile acidic components. The second method is more
accurate because measures
measu are taken to remove these
volatile acids which cofound readings of density calculated
(which is important in determining the % ethanol content
of the wine).
• What is the sulphur dioxide content (free and
combined)
ombined) of each sample of wine?
• What is the total tal acid content of each sample of wine
(TA), assuming that all the acid is tartaric (just as
manufacturers would calculate)?

The selection of wine willl include a red, a white and a rosé.

The red and rosé wines are included for the challenge of decolourising
decolourising them to make some
of the later methods more viable.

Analysing the ethyl ethanol

e content of each wine
Apparatus and Reagents

• 110cm³ of pure ethyl alcohol (ethanol)

• 2 d.p weighing scales
• Distilled water
• 500 cm³ Round bottomed flask
• Thermometer
• Condenser
• Receiver
• 250 cm³ Conical flask

1
 • 250 cm³ beaker
• 100 cm³ graduated pipette
• Measuring cylinder
• Dropper pipette
• Pipette bulb
• pH testing strips
• 200cm³ 0.1 mol sodium hydroxide
• 100cm³ volumetric flask
• Cling film
• Anti bumping granules
• Heating mantle

It is the fine white coating on the surface of ripe grapes (the bloom) which is responsible for
the production of ethanol in the wine. The white coating contains yeasts which can turn the
grape juice, containing sugars, into wine (when the grapes are crushed) (4). The equation
below summarises how sugars in the grapes are converted into ethyl alcohol and carbon
dioxide via the action of yeasts such as Saccharomyces ellipsoideus:

C6H12O6 2 CH3CH2OH + 2 CO2

The first step is to produce a calibration graph which will allow me to make accurate
readings of the ethanol content of my wines. The graph will plot density in grams per cubic
centimetre (gcm¯³) against ethanol as a percentage of the total volume of the solution.

In order to produce this calibration graph I will need to find the density of several different
concentrations of ethanol in solution. I will make 10 solutions of ethanol from 0-20%
ethanol diluted with distilled water. Each solution will be 100cm³ in total volume- therefore
the composition of each solution will be just as illustrated in the table below:

% ethanol in solution Volume of water\ cm³ Volume of ethanol\ cm³

2 98 2
4 96 4
6 94 6
8 92 8
10 90 10
12 88 12
14 86 14
16 84 16
18 82 18
20 80 20

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The density of each solution will then be calculated with the following equation:

Density (gcm¯³) = Mass (g) / Volume (cm³)

Procedure for producing the calibration graph

1) Note down the temperature of the room and make sure that the ethanol being used
is also at this room temperature.
2) Weigh the volumetric flask (with its lid) on the 2d.p. scales and note this down.
3) Add 2cm³ of ethanol to the volumetric flask using a burette then top it up with
distilled water (from a distilled water bottle) to the 100cm³ line on the neck of the
flask.
4) Re-weigh the bottle on the scales and note this down.
5) Empty the contents of the flask and thoroughly rinse the flask with distilled water.
6) Repeat steps 2-4 for the different volumes of ethanol shown in the table above for
volumes of ethanol up to 20cm³.
7) For every reading of mass recorded, the mass of the flask must be taken away from
this figure to get the mass of the solution inside.
8) Density can then be calculated for each solution by dividing the mass of the solution
by 100 (which is the total volume of each solution).

The density of each solution will be calculated and tabulated in the table shown in figure 5 of
the results tables section.

N.B

The recordings required to produce the calibration graph should ideally all be collected in
the same session because then it is more likely that the temperature of the room and that
of the ethanol has not changed as much. The importance of the temperature of the ethanol
will be discussed later in the justification of the first method.

The data from the table will then be displayed in graphical form with density plotted against
% ethanol in solution. A line of best fit should be drawn on the graph; this should be fairly
easy as I would expect a linear relationship. The resultant graph will be the calibration graph
I use to determine the % ethanol of my 3 samples of wine.

Measuring the % Ethanol in Wine

The First Method for measuring the % ethanol in wine
1) Note down room temperature and ensure that all the wine remains at this
temperature as the method is carried out.
2) Transfer the chosen wine to a 250cm³ beaker.
3) Remove 100cm³ of the wine using a 100cm³ graduated pipette.

3
 4) Add this wine to a pre-weighed 250cm³ conical flask
5) Re-weigh the conical flask with its contents. The mass of the conical flask should be
taken away from this figure to give the mass of the solution which should be
recorded in a table.
6) Steps 2-4 should be repeated twice and a mean of the three values for the mass
should be taken.
7) The density of the solution of wine should be calculated using the density equation
discussed earlier.
8) The % ethanol of the wine in question is determined by reading off the y-axis of the
calibration graph the density calculated in step 6 and finding the corresponding %
ethanol content on the x-axis of the calibration curve.

The above should be done for each of the wines.

Justification of the first method for measuring the % ethanol in wine

In order to determine the % ethanol in each sample of wine, the measurements of

wine would need to be to the highest possible accuracy. In the lab the best means of
doing this is with the use of a 100cm³ graduated pipette.

Additionally it was extremely important that the temperature of the wine remains
constant while the method is being carried out; this is because as temperature of the
wine (or any other solution) rises, so does its volume. If the volume of the solution
gets bigger for a given concentration then its density will be smaller (because of the
inverse relationship between density and volume). Therefore the temperature must
be constant while data for calculating density is being recorded.

Scales measuring to 2.d.p were used because in order to calculate an accurate

density for each sample of wine, an accurate measurement of mass for each sample
of wine is crucial.

Furthermore, 2 repeats for the measurement of each wine’s mass were taken to
increase the reliability of my results and reduce the factor of random error in my
experiment.

Limitation of the primary method

The limitation of the primary method is that there are small quantities of volatile acidic
components present in the wine (e.g. acetic acid, formic acid and carbonic acid) which
would contribute to the value of the density calculated in the first method. This leads to
an inaccurate value for the % ethanol in solution. In fact, by using the primary method I
will get a % ethanol value higher than the actual value; this is because the volatile acidic
components have a higher mass than water. For the first method to be valid I would

4
have to assume that the 100 cm³ of wine was only a mixture of ethanol and water;
which is certainly not the case.

Therefore, to overcome this limitation I will use a supplementary method. This second
method should give me a more accurate result for % ethanol content because the
volatile acidic components will be eliminated from my test sample.

The second method

With this method I aim to get as close as I can to getting a mixture of pure ethanol and
water from my sample of wine. In order to do this I am going to remove the ethanol
from a 100cm³ sample of the wine before then adding distilled water to it until it
becomes a 100cm³ solution of ethanol of water. This solution can then be weighed to
find the density and by reading off the calibration graph, an estimate for the %ethanol in
solution can be found.

I am going to remove the ethanol from the sample of wine using the process of
distillation. However, in order to remove the small quantities of volatile acidic
components (weak acids) present in the wine, the wine will have to be neutralised with
alkali beforehand. The volatile acids are a product of microbial metabolism (1). If these
weak acids were able to enter the distillate because of their low boiling temperatures,
the value of the wine’s density will be corrupted.

The total volatile acidity of wine is typically no more than 0.06% by volume, with acetic
acid being the main contributor (2).

0.1 mol sodium hydroxide will be used to neutralise the weak acids present in the wine.
To make the calculation simpler I am going to assume that all the volatile acids present
are acetic acid. The volume of sodium hydroxide needed to neutralise the acetic acid is
calculated below.

5
The calculation suggests that I should use 10.7cm³ 0.1 mol sodium hydroxide to
neutralise the volatile acids present in each sample of wine. However, this is assuming
that all of the volatile acids are acetic acid. An acetic acid molecule is neutralised with
w
the loss of a hydrogen atom which then allows it to turn into an insoluble salt and so
only needs to react with one molecule of sodium hydroxide. On the other hand,
carbonic acid (another volatile acid present in wine) is diprotic which means it would
need to react with twice as many sodium hydroxide molecules (2 molecules) in order to
be neutralised. Therefore,, assuming that all the volatile acid in the wine is acetic acid is
not valid because it will give me a volume of alkali too low to neutralise all
al the volatile
acids.

6
 For this reason I will aim to neutralise the wine by adding twice the amount of sodium
hydroxide calculated in the figure above, 21.4cm³. To confirm that the solution of wine
is indeed neutralised, I will use a pH testing strip.

Oncee the volatile acids in the sample of wine have been neutralised the sample should
then be put in the round bottomed flask of the distillation apparatus with a few anti-
anti
bumping granules. The anti-bumping
anti granules provide nuclei on which gas bubbles can
form
m which leads to smooth boiling rather than bumping,
bumping which can be hazardous. The
solution should be heated and the ethanol should boil out at 72.62⁰C.
72.62 C. The solution
should be allowed to reach 100⁰C
100 C to ensure that all the ethanol has boiled off. The
distillate should then be topped up to 100 cm³ with distilled water in a volumetric flask,
using a burette to add the water with control. The mass of the volumetric
volumetric flask should
be known so that when it is weighed with the solution inside, the mass of the solution
can be ascertained.

Once the mass of the solution is calculated the density can also be found from the density
equation discussed earlier. With a value for density an estimate for the average % ethanol
content in the solution can be found from the calibration graph produced in one of the
earlier stages. Figure 1

thermometer

500cm³ round
bottomed flask condenser

100cm³ volumetric flask

ater out
Water

Water in

Procedure for the second method of finding the % ethanol content of wine
1) Pour more than 100cm³
100cm of the wine into a 250cm³ beaker. Then with a 100cm³
100cm
graduated pipette, 100cm³
100cm of the wine should be removed from the beaker into a
250cm³ conical flask.

7
2) Approximately 22cm³ of 0.1 mol sodium hydroxide (measured using a measuring
cylinder) should then be added to the wine in the conical flask.
3) The pH of the solution must then be tested using a pH testing trip which should
show that the solution is neutral before step 4 is carried out. If the solution of wine
is shown to be still acidic (given by the colour of the pH testing strip), the sodium
hydroxide should be added approximately 2cm³ at a time with a dropper pipette
until a neutral result is obtained.
4) Once the solution is found to be neutral the contents of the conical flask should be
transferred to the round bottomed flask of the distillation apparatus with a few anti
bumping granules.
Note that before the heating mantle is switched on and the process of distillation is started,
the distillation apparatus should be set up correctly as depicted in Figure 1 and cold water
should be allowed to run through the condenser.
5) The heating mantle with the round bottomed flask (with contents) resting in it
should be turned on and the temperature shown on the thermometer should be
monitored and allowed to reach 75⁰C.
6) A distillate should then be collected in a 100cm³ volumetric flask that has been pre-
weighed. To prevent loss of the ethanol collected, cling film should be used around the join
between the spout from the condenser and the opening to the volumetric flask.
7) When the distillate has stopped collecting after approximately 15 minutes, distilled
water should be added to the volumetric flask to the 100cm³ mark. The room
temperature should be measured at this point and the distillate should be allowed to
cool to this temperature before any distilled water is added. Furthermore, this step
along with the following step should be carried out at the same temperature for
each sample of wine.
8) The volumetric flask should then be re-weighed and the mass of the volumetric flask
found earlier should be subtracted from the value given on the scales to find the
mass of the solution. The density of the solution can be found as before and
tabulated.
9) The experiment should be repeated for each type of wine.

Further justifications of the second method of measuring % ethanol in

the wine

Firstly, a measuring cylinder is used to add approximately 22cm³ of sodium

hydroxide because a high degree of accuracy is not required; the base only functions
as a neutralising agent and is not involved in any other reactions.

Furthermore, I have chosen a heating mantle as my heating source over a bunsen

burner for a number of reasons. Firstly, a bunsen burner is an extremely uneven
source of heat, which would lead to the glass at the bottom of the flask being a lot
hotter than the rest of the flask, and undesired reactions might occur. The main
advantages of using a heating mantle are that it is primarily a lot safer because there

8
 is no naked flame; it is also a lot easier to control via its variable
able voltage dial. It is
important that a heating mantle is used because there is no chance of it igniting the
flammable contents of my round bottomed flask if it were to break; the mantle
contains a circuit breaker which cuts the power when any liquids come co into contact
with it (3).

Moreover, I decided that it would improve the accuracy of my investigation if I

collected the distillate directly into the volumetric flask instead of collecting it in a
beaker then transferring it.it. If I were to have collected it in another receptacle before
transferring it to the volumetric flask for weighing, it is likely that I would lose a small
amount of my distillate leading to slight inaccuracies in my results. Additionally, I
found the neck of the volumetric flask to be very narrow, this meant that it was
better than a beaker for limiting the amount of evaporation of my distillate.

Additionally, it is of extreme importance that the distillate of each sample of wine is

made up to 100cm³ in the conical flask at the same temperature; this is for the
reason highlighted in the justification of method 1-
1 changing the temperature of the
solution alters its density.

Finding the Sulphur Dioxide content of wine

Sulphur dioxide is a very important additive in wine because it prevents the presence of
unwanted bacteria and is also an antioxidant. The Sulphur dioxide prevents the wine from
becoming oxidised, which ruins the taste. However, on the other hand too much sulphur
dioxide will also ruin the flavour of wine. Therefore, a fine balance is needed. Figure 2 below
illustrates where the sulphur dioxide added to the wine ends up chemically speaking for
normal wine pH values (3.0 to 4.0).
4.0) Figure 2

The majority of the sulphur dioxide in wine primarily combines with various aldehydic and
ketonic molecules and the rest of the un-combined
un sulphur dioxide
de molecules protect the
wine. This ‘’free’’ sulphur dioxide exists as molecular SO₂,
SO bisulphite
phite and sulphite,
sulph and the pH
of the wine determines how free sulphur
sul ur dioxide is distributed between these three forms.
The aldehydic and ketonic molecules are carbonyl
carbonyl compounds such as sugars (4). A reaction

9
of a carbonyl
rbonyl group reacting with a hydrogen sulphite ion can be seen below to form
addition compounds:

The ‘>’ represents two bonds. If one of these bonds joins to a hydrogen atom, atom the
molecules is described as an aldehyde,
aldehyde If it does not, it is a ketone. Ann aldehyde is a
substance containing the CHO group at the end of the molecular chain. A ketone has the
general formula R(CO)R', where R may be the same as R’. R The equations shown above are
addition reactions forming addition compounds which are also called 'bisulphite addition
compounds'. In fact, to be more specific, the reactions illustrated above clearly illustrate
nucleophilic
ic addition which is a kind of addition reaction which involves the attachment of a
nucleophile to an electron-deficient
deficient part of the
the molecule. Molecules with carbonyl groups
undergo this kind of reaction because of the polarisation of the carbonyl group. In
conclusion, nucleophilic addition reactions occur between sulphur dioxide in the wine and
any carbonyl compounds also in the wine and has the effect of removing ‘free’ sulphur
dioxide and making it ‘bound’.

The ‘free’ sulphur dioxide provides electrons to stabilise free radicals which cause the wine
to spoil. Free radicals are atoms
atom or groups of atoms with at least one unpaired electron.
el

The total sulphur dioxide content of a sample of wine is a measure of the ‘combined’ form
as a complex with some organic compounds as well as the ‘free’ forms such as SO₂
SO (aq),
HSO₃¯(aq)(bisulphite) and SO₃²ˉ(aq)
SO (sulphite).

Dealing with the issue of colour in the red and rosé wines masking the end
point of my titrations Figure 3

For the next two experiments, titrations are carried

out which rely on myself being able to clearly
clearl
distinguish the completion of the reaction with the
abrupt change of colour of solution; this poses a
difficult task with the red and rosé wines which are
heavily coloured, masking the end point of the reaction,
when the colour of the indicator becomes visible.
v

Anthocyanins are the red, blue and black pigments in

grapes which are usually found in the skins of the grapes, giving them their colour. There are
five different
erent anthocyanin compounds found in red wines, the dominant one being malvidin

10
 Figure 4
(shown in Figure 4),, which is red in the acid conditions
of wine. Figure 3 clearly illustrates how the malvidin
present in wine appears differently in protonated
environments (in the left test tube) and in de-
de
protonated environments (in the right test tube).
Although
ugh the test tube on the right is a lot lighter in
colour than the untreated wine, it is still not ideal for
accurate judgement of the end point of my reactions,
which is why I have opted for the activated carbon
method.

In order to remove the anthocyanins from the wine a sample of wine must be treated with
activated carbon in the following method:

1) Add 15 grams of activated carbon/charcoal to 200cm³ of red or rosé wine in a

conical flask.
2) Heat the conical flask over a bunsen burner on low heat for 20 seconds
second
intermittently with a thermometer inside the solution,
solution, removing the conical flask
from the flame every few seconds, swirling the flask gently as it is heated. The wine
must not exceed a temperature of 35⁰C.
35
3) Filter the solution from the conical flask through a
funnel with filter paper into
i another clean conical
flask. Shown in Figure 5.
5

Activated charcoal is charcoal which has been purified and

treated chemically or physically to produce microfissures that
t
increase its adsorptive
sorptive surface area. The charcoal is
“activated” by heating several forms of carbon at high
temperatures in the presence of steam. This process causes Figure 5
the charcoal to form pockets and pores in its main body which increases its surface area.
The large surface area is extremely effective at adsorbing a wide range of polar compounds,
especially polyphenols (such as malvidin) and their derivatives.

The solution is heated to speed up the adsorption process;

process; the molecules gain more kinetic
energy
nergy as the solution is heated and therefore there are more collisions with the carbon
surface were unit time. The swirling of the conical flask makes the carbon disperse
homogeneously in the solution, allowing it to combine/adsorb to the pigments present in
the wine. Furthermore, the wine is carefully monitored to make sure that it does not reach
a high temperature to ensure that volatile acids and volatile sulphur containing compounds
are not lost from the wine; these components of the wine are required to give me accurate
measurements for the total acid quantity and amounts of sulphur dioxide present in the
wine.

11
Apparatus and Reagents
• 370cm³ 4 moldm¯³ sulphuric acid
• 500cm³ 1M sodium hydroxide
• 120cm³ 1% starch solution
• 900cm³ 0.01 M iodine solution
• 250 cm³ conical flask x2
• 50cm³, 25cm³, 10cm³, 5cm³ graduated pipettes and a dropper pipette.
• Pipette bulb

Method for finding free sulphur dioxide content of wine

1) Pipette 50cm³ of the chosen wine into a 250 cm³ conical flask using a 50cm³
graduated pipette.
2) Add 5cm³ of 4 moldm¯³ sulphuric acid using a 5cm³ graduated pipette and 3cm³
of starch solution using a dropper pipette.
The titration is carried out in acidic conditions so the hydrogensulphite ions,
HSO₃¯ react with H+ ions. When these two ions react the sulphur dioxide is
regenerated and this can then go on to react with the iodine, just as it did when
it was ‘free’ to start with (5). The reaction of hydrogensulphite ions reacting with
H+ ions can be seen below:

3) Titrate with 0.01M I₂ (in burettes) until the first blue colour appears. A white tile
should be placed underneath the conical flask before the titration begins.

Method for finding free and combined sulphur dioxide content of wine

1) In a separate 250 cm³ conical flask add 25cm³ of 1M sodium hydroxide using
a 25cm³ graduated pipette.
2) Pipette into the conical flask with sodium hydroxide 50cm³ of wine using a
50cm³ graduated pipette and shake it with a bung in the conical flask.
3) Leave this solution for 15 minutes.

The alkali frees the sulphur dioxide from the organic complexes via the following
reaction:

12
 4) Add 10cm³ 25% sulphuric acid using a 10cm³ graduated pipette and
3cm³starch solution using a dropper pipette and titrate it with more of the
0.01M I₂ in the burette.
5) Titrate with 0.01M I₂ (in burettes) until the first blue colour appears. A white
tile should be placed underneath the conical flask before the titration begins.

For each method of measuring the free sulphur dioxide and fixed sulphur, the methods
should be repeated until I get two titres within 0.1 cm³ of each other.

The reaction showed below shows how the iodine in the burette will react with the
sulphuric acid in the conical flask. The I₂ will be reduced and the SO₂ will be oxidised. (6)

When all the sulphur dioxide has reacted with the iodine, then the iodine will react with the
starch in the solution. The exact mechanism isn’t known but it is thought that the Iodine in
the form I₃¯ and I5¯ ions fit inside the coils of amylose in starch. The energy level spacing in
the resulting complex corresponds to the absorption spectrum in the visible light area in
our electromagnetic spectrum. The colour of the complex formed appears blue. Therefore
any excess Iodine added to the solution which can not react with any of the sulphur
(because it has already reacted) reacts with the iodine forming a sharp blue complex.

Therefore, when this blue colour is seen in the titration, I will know that all of the sulphur
dioxide in the wine has reacted with the Iodine and I can stop titrating the iodine. However,
there is a problem that the red colour in red wine will mask this blue colour produced
when the iodine reacts with the starch. In order to remove as much of this red colour from
the wine as is possible to improve the chances of seeing the colour change of the solution,
activated charcoal will be used as described in the method on page 9.

In order to find the total amount of sulphur dioxide present in parts per million
in the sample of wine the total number of cm³ of iodine is multiplied by 12.8 (7).

Justification of method to find sulphur dioxide content

Graduated pipettes are used to measure out the chemicals in solution in the conical flask
because they are the most accurate way of measuring volumes of my chemicals. If my
volumes of reactants are measured with accuracy, then I am more likely to get titre values
of iodine which are more accurate. Using the graduated pipettes adds to the validity of this
part of the investigation because of this.

Furthermore, repeat titres are carried out until I get two concordant titres within 0.1 cm³
because this adds to the reliability of the investigation which leads to improved accuracy in

13
my results. Iodine is also added to the conical flask from a burette because this enables me
the greatest possible control over the rate at which the chemical is added to the solution.

Adjustments to method
While performing the titration of Iodine for my method I noticed I was getting very small
titres of Iodine. I thought that I could improve the accuracy of my results by using a lower
concentration of Iodine, which would give me larger titres of the Iodine. This is because in
order to determine the percentage uncertainty of my titre of iodine, the absolute
uncertainty of the burette is divided by the measured volume of Iodine before being
multiplied by 100; therefore, the larger the volume of iodine, the lower the percentage
error of my measurement.

The new concentration of iodine used was 10 times less concentrated than the original
iodine used, 0.001 moldm¯³. Therefore, In order to calculate the amount of SO₂ in the
sample of wine, the volume of Iodine added in cm³ is multiplied by 1.28 instead of 12.8.

14
15
Finding the total acid content of wine (the sum of the
titratable acids when the wine is neutralised)
Wine is generally quite acidic, having a low pH because it contains a mixture of weak
organic acids such as tartaric and malic acid. The weak organic acids are generally derived
from grapes. The acid which makes up the majority of the weak organic acids in the wine is
tartaric acid, however. Because most of the acid in wine is tartaric, most food standards
agencies define the total acidity (or TA) as the total amount of tartaric acid in the bottle of
wine as a percentage. 1.0% is generally considered a high TA level whereas 0.4% would be
too low. Most red wines generally have a TA of of 0.6% which is what I expect to see when
I calculate the total acid content of my wine, assuming all acid is tartaric acid. (8) (9)

The acidity is not normally removed from the wine by the manufacturer because it provides
the enjoyable ‘crisp’ taste of the drink.

My method involves a titration with sodium hydroxide, which reacts with the acid. When
the reaction is over, the sodium hydroxide reacts with the indicator added to the solution.
The wine solution should remain unchanged by the addition of the indicator because it
remains colourless in acid conditions. However, the indicator turns pink in a basic solution.
When I add sodium hydroxide to the wine- a pink colour will appear but should fade as the
weak organic acids in the wine react with the sodium hydroxide (alkali) in a neutralisation
reaction (producing a salt and water). When there is no longer any acid to react with the
base, the base will be able to react with the indicator and the indicator will turn pink
permanently.

The reaction between tartaric (representative of all the weak organic acids in the wine) and
aqueous sodium hydroxide can be seen below:

Na2C4H4O6+2H2O
H2C4H4O6+2NaOH

The equation above illustrates the idea that 2 moles of sodium hydroxide react with one
mole of acid to form one mole of salt and 2 moles of water.

On the subject of phenolphthalein, it is itself a weak acid, and behaves like an indicator
because its conjugate acid and base forms of the molecules have different colours.
Phenolphthalein can lose protons in solution and the molecule itself is colourless. However,
when the molecule is ionised it turns pink, so when a base is added to the solution the
‘molecule ⇌ ions’ equilibrium shifts to the right leading to more ionisation of phenolphthalein
molecules as protons are removed (10).

16
Apparatus and Reagents

• 250 cm³ Conical flask

• 250cm³ beaker
• Burette and stand
• 10cm³ and 5cm³ graduated pipettes
• Plastic dropper pipette
• 300cm³ 0.1Mol Sodium Hydroxide
• Phenolphthalein indicator 1%
• Pipette bulb

N.B
Decolourised wine should be used; produced from the method on pages 10-11

1) Measure 15cm³ of wine into a conical flask with a 10cm³ graduated pipette then a
5cm³ graduated pipette
2) 3 drops of phenolphthalein should be added to the wine in the conical flask with the
plastic dropper pipette.
3) The burette (in its stand) should then be filled with the sodium hydroxide and added
to the solution in the conical flask very slowly until the pink colour remains visibly in
the conical flask. A white tile should be placed underneath the conical flask before
any titrating begins. The amount of reagent added to the solution must be noted and
the experiment repeated until 2 titre readings which coincide within 0.1cm³ are
achieved.

Moles= concentration (moldm¯³) x volume (dm³)

From the equation above, the moles of sodium hydroxide added to the wine can be
calculated. This value should then be halved to find the number of moles of acid in
the wine (seeing as 2 moles of NaOH react with 1 mole of acid). 0.015dm³ of wine
was used, therefore I can work out the concentration of the acid in the sample using
the same equation. This value for concentration can then be converted to gdm¯³
which is then converted to % by dividing by 10.

Justification of method to find total acid content of wine

A white tile is placed underneath the conical flask containing the wine and indicator so that the
sharp end point of the reaction can be recognised quickly, and therefore with greater accuracy. The
pink of the indicator stands out best against the white background of the tile.

Furthermore my method includes repeats to ensure that my results are reliable and greatly reduce
the chance of random error; this makes the method more accurate.

17
 The Risk Assessment
Assessm
Chemical Concentration Hazard Precautions

Ethanol 100% Ethanol is EXTREMELY Safety glasses and good

FLAMMABLE and will readily ignite ventilation are essential.
with heat. Causes skin and eye The ethanol should also
irritation. Ingestion can cause be kept away from any
nausea, vomiting and inebriation; flames or hot objects.
chronic use can cause serious liver
damage. Inhalation of the vapour
can also produce a narcotic effect.

Sodium 0.1 moldm¯³ There is no real risk of 0.1 moldm¯³ Safety glasses should be
Hydroxide sodium hydroxide being corrosive worn at all times to
to the skin, although it is classed asprevent the chemical
an IRRITANT. The chemical is also coming into contact
an irritant if it enters the eyes and with the eyes. If the
swallowing the chemical can do chemical was to come
serious harm to organs in the body. into contact anywhere
on the body it should be
rinsed off immediately
with water; clothing
which has come into
contact with the
chemical should also be
removed. If swallowed,
the mouth should also
be rinsed thoroughly
with water.
Phenolphthalein 1% May act as a skin, eye or respiratory Safety glasses should be
irritant. Animal experiments indicate worn as with all the
a theoretical risk that this material chemicals. The chemical
may be carcinogenic in humans if should be washed off
ingested at high levels over a long with water if it is spilt
period. on skin.

18
Sulphuric Acid 4.0 moldm¯³ This chemical is of a sufficient
concentration to be CORROSIVE
to the skin, eyes and digestive tract.
It is slightly hazardous in case of
inhalation (lung sensitizer) but non-
corrosive for lungs. The liquid or
spray mist may cause tissue damage
to the mucous membranes of the
eyes mouth and respiratory tract.
Skin contact may cause burns or
small blisters.
Sodium 1.0 moldm¯³ This chemical is CORROSIVE to the
Hydroxide eyes and skin. It may also cause
gastrointestinal discomfort and if
inhaled it may cause irritation to the
respiratory tract.
Iodine solution 0.01/0.001 Iodine is HARMFUL in the body if it
moldm¯³ is inhaled or swallowed; it can also
be absorbed through the skin.
However, because it is in a very low
concentration, the risks are
reduced.
Starch solution 1% This chemical is a very low hazard
to the body but could potentially
irritate the delicate membranes of
the eyes.
Distilled water N/A No risk.
19
Safety glasses and latex
gloves should be worn
while handling this
product. If it comes into
contact with the skin,
the skin should be
flushed with water for at
least 15 minutes while
removing any
contaminated clothing.
The eyes should also be
flushed with water if
they come into contact
with it.
Same precautions should
be taken as with the 0.1
moldm¯³ concentration
even though it is 10
times the concentration-
(it is still a very low
concentration).
Care should be taken
when pouring the iodine
into the burette for
titrating. Safety glasses
should be worn and
polyethylene is a suitable
material for gloves if
they are needed.
This should be kept
away from the eyes and
safety glasses should be
worn at all times.
No precautions need to
be taken.
The Results
At 21⁰C- Data for the production of calibration curve of density against % ethanol Figure 6

mass of solution \g volume\ cm³ density \ gcm¯³ % ethanol

99.54 100 0.9954 2
99.29 100 0.9929 4
98.86 100 0.9886 6
98.59 100 0.9859 8
98.32 100 0.9832 10
98.00 100 0.9800 12
97.79 100 0.9779 14
97.51 100 0.9751 16
97.24 100 0.9724 18
96.88 100 0.9688 20

Mass of volumetric flask used- 47.89g

Data for the production of calibration curve of density against % ethanol (Temperature not
monitored)

Figure 7

mass of solution \g volume\ cm³ density \ gcm¯³ % ethanol

98.30 100 0.9830 2
98.48 100 0.9848 4
98.28 100 0.9828 6
97.82 100 0.9782 8
97.72 100 0.9772 10
97.50 100 0.9750 12
100.94 100 1.0094 14
100.77 100 1.0077 16
100.51 100 1.0051 18
100.17 100 1.0017 20

20
 At 21⁰C- Calculating the density of each type of wine

Figure 8
Trial 1 Trial 2\ Trial 3\ Mean Mass of Solution\ volume\
Type of Wine \g g g g cm³ density \ gcm¯³
RED 98.95 99.00 98.96 98.97 100 0.9897
WHITE 98.94 98.89 99.11 98.98 100 0.9898
ROSÉ 98.89 99.88 98.87 99.21 100 0.9921

Mass of conical flask used- 209.27g

Figure 9
At 21⁰C- Mass of distillates collected from distillation and corresponding densities

Type of
Wine Mass of distillate solution\ g Repeat \ g Mean mass\ g volume\ cm³ density \ gcm¯³
RED 98.90 98.87 98.89 100 0.989
WHITE 98.65 98.70 98.68 100 0.987
ROSÉ 98.54 98.50 98.52 100 0.985

Mass of volumetric flask used- 47.89g

A comparison of the different results for ethanol content for each type of wine
Figure 10

Type of Wine % Ethanol with method 1 % Ethanol with method 2

RED 5.693 6.177
WHITE 5.624 7.559
ROSÉ 4.035 8.941

Figure 11
Amount of Sodium Hydroxide base required to neutralise each type of wine

Type of Wine Amount of NaOH used \cm³

Rough 1st 2nd 3rd Mean titre
Start Finish Start Finish Start Finish Start Finish \ cm³
RED 0.00 10.40 0.00 9.60 0.00 9.50 0.00 NA 9.55
WHITE 0.00 11.50 0.00 11.10 0.00 10.90 0.00 10.80 10.85
ROSÉ 0.00 10.80 0.00 10.50 0.00 10.70 0.00 10.40 10.45

21
 Type of Moles of Moles of tartaric
Wine Mean titre of NaOH\ dm³ NaoH acid Figure 12 acid
% tartaric
RED 0.00955 0.00096 0.00048 0.4775
WHITE 0.01085 0.00109 0.00054 0.5425
ROSÉ 0.01045 0.00105 0.00052 0.5225
How moles of Sodium hydroxide corresponds to moles of acid in the wine and therefore the % acid
present

Titres of 0.01M Iodine for each type of wine- used to find free Sulphur Dioxide content
Figure 13

Titre value of 0.01M I₂ for free SO₂ \ cm³

3rd Mean
Type of Wine 1st Titre 2nd Titre Titre titre
Start Finish Start Finish Start Finish \ cm³
RED 0.00 0.10 0.00 0.10 Na Na 0.10
WHITE 0.00 0.20 0.00 0.35 0.00 0.40 0.38
ROSÉ 0.00 0.05 0.00 0.20 0.00 0.15 0.18

Titres of 0.01M Iodine for each type of wine- used to find free and combined Sulphur Dioxide
content
Figure 14

Titre value of 0.01M I₂ for free and combined SO₂ \ cm³

3rd Mean
Type of Wine 1st Titre 2nd Titre Titre titre
Start Finish Start Finish Start Finish \ cm³
RED 0.00 0.50 0.00 0.30 0.00 0.40 0.35
WHITE 0.00 5.50 0.00 5.80 0.00 5.70 5.75
ROSÉ 0.00 2.50 0.00 2.10 0.00 2.20 2.15

22
 How the titre values of 0.01M Iodine correspond to the amount of ‘free’ and ‘free and combined’
Sulphur Dioxide
Figure 15

Type of Mean titre for free Corresponding Corresponding

Wine SO₂ amount Mean titre for free amount
and combined SO₂ \
\ cm³ of SO₂\ ppm cm³ of SO₂\ ppm
RED 0.10 1.28 0.35 4.48
WHITE 0.38 4.86 5.75 73.60
ROSÉ 0.18 2.30 2.15 27.52

Titres of 0.001M Iodine for each type of wine- used to find free Sulphur Dioxide content

Figure 16

Titre value of 0.001M I₂ for free SO₂ \ cm³

3rd Mean
Type of Wine 1st Titre 2nd Titre Titre titre
Start Finish Start Finish Start Finish \ cm³
RED 0.00 2.10 0.00 2.30 0.00 2.15 2.13
WHITE 0.00 3.70 0.00 3.75 NA NA 3.73
ROSÉ 0.00 1.35 0.00 1.55 0.00 1.45 1.40

Titres of 0.001M Iodine for each type of wine- used to find free and combined Sulphur Dioxide
content
Figure 17

Titre value of 0.001M I₂ for free and combined SO₂

3rd Mean
Type of Wine 1st Titre 2nd Titre Titre titre
Start Finish Start Finish Start Finish \ cm³
RED 0.00 3.90 0.00 4.25 0.00 4.00 3.95
WHITE 0.00 57.55 0.00 57.30 0.00 57.40 57.35
ROSÉ 0.00 21.10 0.00 21.10 0.00 2.20 21.10

23
 How the titre values of 0.01M Iodine correspond to the amount of ‘free’ and ‘free and combined’
Sulphur Dioxide
Figure 18

Corresponding Corresponding
Type of Wine Mean titre for free SO₂ amount Mean titre for free amount
and combined SO₂ \
\ cm³ of SO₂\ ppm cm³ of SO₂\ ppm
RED 2.13 2.73 3.95 5.06
WHITE 3.73 4.77 57.35 73.41
ROSÉ 1.40 1.79 21.10 27.01

Analysis and Evaluation

Analysis Figure 19

10

7
% Ethanol by Volume

6 % Ethanol with method 1

% Ethanol with method 2
5

0
RED Type of Wine WHITE ROSÉ

24
 Calibration Curve of Density of Ethanol solutions against % Ethanol in Solution 21⁰C

Figure 20

25
 Calibration Curve of Density of Ethanol solutions against % Ethanol in Solution- Temperature not
monitored
Figure 21

26
The first time the method for producing the calibration graph was carried out, I collected
some very disappointing results which are shown in figure 7. The graph of these results can
also be seen above in figure 21, looking at the blue line. This erroneous calibration graph
shows the opposite trend I was expecting, with density increasing with increased
concentration of ethanol; this observation alerted me to the fact that something was wrong
because data tables told me that ethanol was less dense than water. Therefore, given two
solutions each made up of two chemicals of different densities, the solution with the greater
proportion of chemical with the lower density will have a lower density overall than the
other solution. The actual reason for the opposite trend I was expecting is due to the
solutions containing concentrations of ethanol from 14-20% being collected on a different
day, when either the ethanol or room temperature was a lot cooler. The ethanol being at a
cooler temperature reduced the volume of the solutions relative to the previous solutions
made up, resulting in higher density readings (because density is inversely proportional to
volume). Further evidence for the blue line on figure ? not displaying the correct trend is
that there is a huge amount of scatter from the forced line of best fit. When I draw separate
lines of best fit (shown with red lines) for the data from 2-12 % ethanol and then 14-20%
ethanol concentrations, the correct correlation/trend is seen.

However, on discarding these results and starting again, I made sure that I noted down the
temperature at the start of the method and made sure that I worked efficiently enough to
collect all of my results for the calibration graph in one session, in order to ensure that all
solutions were weighed under the same conditions. As discussed in the method for
producing the calibration graph, fluctuations in the room temperature bring about changes
in the volumes of the solutions. Changes in the volumes of solutions influence the density of
them which is not desired because I wanted a calibration graph with as little spread as
possible. The second attempt at producing a useable calibration graph was much more
successful with very little scatter and showed perfectly the correlation I was expecting.

From Figure 18 above it is clear that the first method for measuring the % ethanol gave
quite different results to the second method, while both methods produced results
substantially below the ‘true’ % ethanol values on the labels of the bottles, (11% for all
three). Looking at the first method, the red wine contained the most ethanol with 5.693%
ethanol followed by the white with 5.624% and then the rosé with 4.035%. However, the
opposite rankings occurred with the second method with the red wine achieving 6.177%,
the white with 7.559% and the rosé with the most at 8.941%.

At 21⁰C (the temperature the first method was carried out at) the ethanol present in each
sample of wine has a density of 0.78860g/cm³ compared to the water present which has
0.99799g/cm³. The density of water is much higher than the density of ethanol at this
temperature; therefore, given two solutions composed of both ethanol and water, the
solution with the higher proportion of water will be denser. From my calibration graph
shown in figure ? I know that high density of solution corresponds to low % ethanol content,
therefore solutions with higher ratios of water to ethanol will have lower % ethanol
content. However, in the samples of wine weighed in the first method it is not just a

27
solution of ethanol and water, there are numerous other compounds present, each with
their own specific density which would influence the % ethanol values (which are
determined by a calibration curve taking into account only solutions of ethanol and water).
Comment on the limitations of this first method will be shown in the evaluation. Figure 22

% Ethanol with %
Type of Wine method 1 % Ethanol with method 2 Difference Increase
RED 5.693 6.177 0.484 8.502
WHITE 5.624 7.559 1.935 34.406
ROSÉ 4.035 8.941 4.906 121.586
Mean % Increase: 54.831

The second method produced results for % ethanol substantially above the % ethanol values
from the first method. The mean % increase from the % ethanol values from the first
method to the second method is 55% which is very substantial indeed (shown in Figure ?).
The reason for this difference is that the number of different compounds in my solution of
ethanol and water is largely reduced due to the distillation process. Volatile weak acidic
components present in the wine are neutralised by alkali to prevent them boiling out at the
same time as the ethanol and entering the distillate corrupting the density value for each
sample.

Additionally, regarding the second method I observe how the theoretical value of % ethanol
stated on the bottles of each wine (11% on all three) are still not reached. My explanation
for this is that between 21⁰ (room temperature) and 75⁰C (the temperature the solution
inside the round bottomed flask of the distillation apparatus was allowed to reach) many
other compounds apart from ethanol and water vaporised and condensed as part of the
distillate. These additional compounds would increase the density of the solution made up
to 100cm³ which was the cause of lower % ethanol values for the second method. However,
the distillation process did prevent a large enough portion of ‘foreign’ molecules from
entering the distillate to show a visible difference in density/ % ethanol between the second
method and first method.

Moreover, I notice that the difference in the % ethanol values between the first and second
methods becomes greater from the red wine to the white wine to finally the rosé wine and
moreover; while the red wine showed to have the greatest amount of ethanol from the first
method, the second method shows it to have the least of the three wines. An explanation
for this discrepancy is that the first method is not accurate at all. I have reached this
conclusion because wine contains such a vast number of different compounds containing
roughly 250 of them, more than in blood serum (11); each compound has its own specific
density at room temperature, therefore measuring the wine’s density to measure the %
ethanol is not likely to give you a true picture of the alcohol content of the wine. The more
compounds present in the wine, the less accurate a measure of density will be for
determining the % ethanol, simply because the ethanol present in the wine represents a
smaller fraction of the total number of different molecules in that sample.

28
 Figure 23

The total % Acid Present in Each Wine

0.56

0.54
% Acid (assuming all tartaric)

0.52

0.5 RED
WHITE
0.48
ROSÉ

0.46

0.44
RED WHITE ROSÉ
Type of Wine

29
 Figure 24

30
In terms of the acidity of each wine there was quite some variation between the three
wines. While the red wine showed the least total acid content with 0.4775% the rosé
appeared to have a fair bit more with 0.5225% while the white wine had the most with
0.5425%. With a typical table wine having a total acid content of 0.6%, the results I collected
are not very far off this guideline value at all; this is probably because the tartaric acid (which
is what I was measuring) is not a volatile acid (2). If the acid I was measuring was volatile
then it would be more likely for me to have measured total acid values significantly below
the typical amount because of the heating process in the decolourisation of the wine
method: the heating of the wine would have allowed the volatile acid to evaporate when
heated and because the tartaric acid is such a major constituent of the total acid content,
the total acid value would have been significantly reduced. On the other hand, acetic acid is
a constituent of the Volatile Acids present in wine. A fair amount of the acetic acid in the
wines would have been lost in the process of decolourising the wine, however because the
volatile acid content of wine is never normally more than 0.06% this would not have affected
my results to any significant extent.
Figure 25

The Free and Combined Sulphur Dioxide Content

for Each Wine
80.00

70.00
Amount of Sulphur Dioxide\ppm

60.00

50.00

40.00
Amount of SO₂\ ppm from 0.01 I₂
30.00 Amount of SO₂\ ppm from 0.001 I₂

20.00

10.00

0.00
RED WHITE ROSÉ
Type of Wine

31
 Comparison of the 'free' portion of Sulphur
Dioxide against the 'combined' portion
80.00

70.00
Amount of Sulphur Dioxide\ppm

60.00

50.00

40.00
Amount of combined SO₂\ ppm
30.00
Amount of free SO₂\ ppm
20.00

10.00

0.00
RED WHITE ROSÉ
Type of Wine

Figure 26

The full step by step method/workings for calculating the amount of sulphur dioxide in each
sample of wine can be seen on page 15. Like the total acid content results, the total sulphur
dioxide content results were similarly quite varied between the wines. Both concentration
of iodine produced similar results for the amount of sulphur dioxide in each wine, however
the 0.001 molar iodine produced more accurate results for the quantity of sulphur dioxide
in each wine because of its larger titres; therefore I shall describe the results from titrating
with the 0.001 molar iodine. The red wine showed the least amount of sulphur dioxide with
5.06ppm while the white showed to have roughly 15 times more with 73.41ppm and the
rosé wine had a quantity of sulphur dioxide closer to the value of the red with 27.01ppm.

The results showed lowest amounts of sulphur dioxide in the red wine and highest in the
white, with rosé wine somewhere between. This is explained by the fact that red wines do
not need any added sulphur dioxide because they naturally conatin anti-oxidants (acquired
from their skins and stems during fermentation); although winemakers often add some
anyway. White wines and Rosés typically contain a lot more because they do not contain
high enough amounts of natural anti-oxidants because they are not left in contact with their
skins after crushing. Therefore, white and rosé wines are more prone to oxidation and tend
to be given larger additions of sulphur dioxide by the manufacturers (12).

A method was described in the planning section for measuring the ‘free’ sulphur dioxide
present in each sample of wine as an extension of the original question at the start of the
plan. Originally I did not think that the ‘free’ sulphur dioxide in wine was of any great
significance because EU guidelines on the amount of sulphur dioxide base figures on the

32
‘free and combined’ sulphur dioxide. Additionally, manufacturers of wine typically look at
the total amount of sulphur dioxide because it is ‘free’ as well as ‘combined’ sulphur dioxide
which contributes towards the taste of the wine. I decided after producing my plan that the
amounts of ‘free’ sulphur dioxide are in actuality of most importance. The ‘free’ sulphur
dioxide is the constituent which ionises in water and is the active form of sulfur dioxide
which deactivate microbial enzymes, removes free oxygen from wine (which causes it to
spoil) and kills microbes (13). Therefore, I wanted to confirm that not all of the sulphur
dioxide had combined with organic compounds to form complexes and that ‘free’ forms
were indeed present to offer some protection to the wine.

From the graph I produced in Figure ? I can clearly see how ‘free’ sulphur dioxide is present
in all three bottles of wine. The white wine contained the most with 4.77ppm which is what
I would expect because of it lacking natural antioxidants; the red contained 2.73ppm of
sulphur dioxide and the rosé contained 1.79ppm. Additionally, I notice how with the white
and rosé wines, most of the sulphur dioxide is in the ‘combined’ state. (The red wine does
not follow this trend because of the naturally high antioxidant content discussed earlier.)
This is because sulphur dioxide is reactive and will gradually combine with aldehydic and
ketonic organic molecules to form complexes; the proportional difference between ‘free’
and ‘combined’ sulphur dioxide is not seen as clearly in the red wine because very little
sulphur dioxide is added to this type of wine in the first place.

Evaluation
% uncertainties in measurements
Total % uncertainty in values of density for calibration graph-

0.005/144.77 X 100= 0.00345% (3sf) percentage error of mass reading of flask with
contents

I have used the lowest mass recorded in the table of results for the calibration graph to give a worst
case scenario.

0.005/47.89 X 100= 0.0104% (3sf) percentage error of determining mass of empty

volumetric flask

0.15/100 X 100= 0.15% percentage error of volume reading

[(0.05/2) X 100] X 2 = 5% percentage error of measuring 2cm³ of ethanol from

burette

Total percentage uncertainty in density reading = 5.16% (3sf)

It is clear from the broken down calculation of total percentage uncertainty that the
greatest source of error in my density readings comes from measuring out quantities of

33
ethanol using the burette. In hindsight it would have been more accurate to use different
sized graduated pipettes for these measurements. Furthermore, a larger volumetric flask
(e.g. 250 cm³) would have reduced the error quite substantially for the calibration curve and
in retrospect I should have done this. The same would have reduced the uncertainty in
density readings for the first method of determining the % ethanol of the wine, but at the
time of planning my investigation I considered this method to waste an awful lot of wine. It
could be argued that the wine weighed in the first method could be reused; however, for
the sake of preventing as much of the volatiles from escaping as possible and any possible
contamination, I decided that discarding would be the best option.

The mass readings of the volumetric flask with contents and empty volumetric flask have so
little error associated with them and contribute so little to the total % uncertainty that I will
not comment very much on them. However, what I will say is that if there was a four
decimal place set of scales available, this would have reduced the % error by a great factor.

Total % uncertainty in values of density from First Method

0.005/209.27 X 100= 0.00239% (3sf) percentage error of determining mass of empty

conical flask

0.12/100 X 100= 0.12% percentage error of 100cm³ graduated pipette measure

0.005/308.14 X 100= 0.00162% (3sf) percentage error of determining mass of

conical flask containing wine

Total % uncertainty in values of density from First Method= 0.124% (3sf)

There is a massive fall in the % uncertainty from the calibration graph method to the first
method involving the wine; this is mostly due to the fact that the burette was not used to
measure out small volumes of chemicals. The largest influence on the total % uncertainty
was the use of the graduated pipette. I do not believe that there was a more accurate way
for me to measure out 100cm³ of wine except from using a higher ‘class’ of glassware.

Total % uncertainty in values of density from Second Method

0.12/100 X 100= 0.12% percentage error of 100cm³ graduated pipette measure

0.15/100 X 100= 0.15% percentage error of volumetric flask

0.005/146.39 X 100= 0.00342% (3sf) percentage error for mass of distillate

0.005/47.89 X 100= 0.0104% (3sf) percentage error for mass of volumetric flask

Total % uncertainty with density quantities = 0.284 % (3sf)

Like the first method for determining the % ethanol the second method has relatively low %
uncertainty associated with the final quantity, although it is slightly more than double the

34
uncertainty from the first method. The reason for the increase from the first method was
because I had to make up the solutions to be weighed in volumetric flasks each time, which
adds an additional 0.15% to the total uncertainty. This could have been improved by using a
larger volumetric flask (e.g. 250cm³); however, because the volumes of my ethanol distillates
were so small and didn’t vary very much in size, I would have got a larger range of results
from using a 100cm³ volumetric flask, which increases the validity of this method.

Total % uncertainty in values of % tartaric acid

0.05/10 X 100= 0.5% percentage error of 10cm³ graduated pipette measure

0.025/5 X 100= 0.5% percentage error of 5cm³ graduated pipette measure

[(0.05/9.50) X 100] X 2 = 1.05% (3sf) percentage error of burette reading

Total % uncertainty in values of % tartaric acid = 2.05% (3sf)

This part of the investigation appears to have midrange % uncertainty associated with the
final % tartaric acid values, compared with the other calculated total % uncertainties. The
lowest titre of sodium hydroxide was used to give a worst case scenario for the percentage
error of the burette readings. Like other parts of the investigation it is usually (with the
exception of the sulphur dioxide method) the burette which contributes most to the total %
uncertainty. This percentage error was unavoidable with the choice of equipment available
to me as the burettes in the lab have a precision error of ±0.05. Doing some research
online I found ‘Class A’ burettes with precision errors of ±0.03 which would reduce the
percentage error by a factor of nearly 2; however, sadly I did not have access to such
burettes.

Total % uncertainty in total (free and combined) quantities of Sulphur Dioxide

\ppm

0.05/50 X 100= 0.1% percentage error of 50cm³ graduated pipette measure

0.05/25 X 100= 0.2% percentage error of 25cm³ graduated pipette measure

0.05/10 X 100= 0.5% percentage error of 10cm³ graduated pipette measure

0.25/3 X 100= 8.33% percentage error of 3cm³ measured from dropper pipette

[(0.05/3.90) X 100] X 2 = 2.56% (3sf) percentage error of Iodine titre

Total % uncertainty in total quantities of Sulphur Dioxide \ppm= 11.69%

From these calculations it is easy to see how the dropper pipette influenced the large total
% uncertainty the most contributing 8.33%, followed by the titration procedure contributing

35
2.56%. In order to lower this total % uncertainty I should have used a 5cm³ graduated
pipette and filled it to the 3cm³ line with indicator. Higher quality graduated pipettes (Class
A instead of Class B) would have also improved the total % uncertainty, however these
were not available to me.

Finding % ethanol content of each wine

The first step of this part of the investigation required the production of a calibration graph.
The main limitations of my procedure were fluctuation in temperature, which would affect
the volumes of my solutions and my ability to measure the liquids accurately. Furthermore,
it was brought to my attention during my practical that the ethanol I was using was not
100% pure; instead it was supposedly 95% pure, this would mean that my standard solutions
would have density values higher than the theoretical values if I was using 100% pure
ethanol. Ethanol is less dense than water, which is why this is the case. Therefore, later
when using the calibration graph to match density with corresponding % ethanol, the %
ethanol indicated by the graph will be an underestimate of the true value of the % ethanol.

However, I can say that the calibration graph is a reliable product of the method because
there is very little scatter about the line of best fit; the density values for each different
solution fit the linear trend/relationship very well. This shows that the method was carried
out with very little random error and measurements were made with great accuracy. Even
though the ethanol was not at the ideal concentration, the graph produced will still work for
determining the relative differences in % ethanol between the samples of wine. Additionally,
it should be noted that the error in the density values from the calibration graph are
extremely small. The largest solution of ethanol made for the graph is “20 % ethanol” which
in actuality is only 19% ethanol when using a 95% solution of ethanol. The actual difference
in density between the “20% ethanol solution” produced from my 95% ethanol and the
theoretical value for the density of a solution made with 100% ethanol is only 1.33%; this is
not of any great significance. The calculation for this figure can be seen on the next page in
Figure 27.

36
 Figure 27

37
The chemical reasoning behind the second method producing % ethanol values considerably
below those stated on the bottles is that during the distillation process, not all the ethanol
boiled out of the solution. Even though plenty of time was left for the ethanol to vaporise
and the temperature was high enough for it to do so, the actual amount collected was less
than expected; possibly because the anti-bumping granules kept bouncing and making big
bumps causing me to stop heating for a short while to prevent the bumping. Additionally I
occasionally had to stop the distillation momentarily before the solution inside reached
100⁰C to prevent explosion of the round bottomed flask vessel. These variations in
temperature coupled with the fact I had to stop the distillation before it reached the boiling
point of water may have been the cause of incomplete distillation of ethanol. Moreover,
after I had collected my results I noticed that the way I was storing my bottles of wine was
encouraging the loss of ethanol from the solution between the bottle and the loose lid of
the bottle which was stood upright. Ethanol is very volatile which enables it to evaporate
out of the bottle through any cracks at low temperature. However, if the bottles were to
have been stored horizontally the loss of volatile ethanol would have been significantly
reduced.

Measuring the total sulphur dioxide content of wine

As mentioned before a lower concentration of Iodine was used for a second set of titrations
against each solution of wine containing sulphur dioxide in order to give larger titre values
and therefore greater accuracy in determining the amount of SO₂ in each sample. The main
limitation of the procedure was that in the process of decolourising the red and rosé wines,
volatile sulphur containing compounds would have been lost from the sample making the
total sulphur dioxide content of each lower than their true values. Compounds such as
hydrogen sulphide and ethyl mercaptan are a couple of the volatile sulphur containing
compounds often found in wine which would have been lost in part by the light heating in
the decolourisation process. This limitation could have been greatly reduced by emitting the
heating in the decolourisation of the wines. By leaving the wine to stand once the charcoal
had been added it is likely that the colour from the samples of wine would be reduced.
However, this would be very time consuming and because time did not allow, I was obliged
to catalyse the adsorbing process via some light heating.

Additionally another limitation of the procedure is that when titrating with a white tile
underneath the conical flask, it is still challenging to determine the end point of the reaction.
To make this judgement of the end point of the reaction easier I think that I could
overcome this problem by using a yellow tile. I believe a light yellow tile would make the
dark blue colour produced by the reaction between the iodine and the starch stand out
more vividly than if I was using a white tile: this would make the judgement of the endpoint
of the reaction more accurate. Yellow is the complimentary colour to dark blue which is
why I have come to this innovative solution.

Furthermore, another limitation was that the method for measuring the sulphur dioxide
content of each wine was carried out two weeks after the bottles were opened. This meant

38
that ‘free’ sulphur dioxide could have potentially been lost from each bottle because SO₂ gas
would have escaped from each bottle into the atmosphere each time a bottle was opened.
This limitation could have been overcome by doing this part of the investigation in the first
week, immediately after decolourising the wine. However, in practice the timing of lab
sessions did not allow for this. A more creative way of overcoming this limitation would be
to in the first week transfer all the wine from each bottle into its own squeezable plastic
bottle with valve cap. A two litre plastic bottle water is sold in with a sports drink bottle cap
(the valve type) screwed on would be a good way of doing this. This way when a sample of
wine is required, instead of unscrewing the wine cap and letting SO₂ gas escape, the plastic
bottle can be upturned (so only liquid and no gas comes out), and squeezed to get the wine
into whatever receptacle is chosen.

Measuring the total % acid content of wine

Like the limitations of the sulphur dioxide method, the main limitation of this method was
that the red and rosé wines used had been subjects to the decolourising process, which
would have driven off a portion of the volatile acids measured by the method. This would
result in an underestimate in the total acidity percentage calculated from the data recorded.
To overcome this limitation of the titration method, (which requires clear solutions in order
to effectively determine the end point of the reaction) I could have used a digital pH metre
which I would be zeroed before immersing in solutions which hadn’t been decolourised. The
use of a pH metre would have given me a clear idea of the relative acidity of all three types
of wine; however it would not have given me any indication of the % total acid so it is not an
ideal solution. Emitting the heating in the decolourisation process would be a better solution
however, the adsorption process would inevitably take a lot longer at room temperature.

Additionally, the percentage uncertainty value of each total % acid could have been reduced
by using a single 15cm³ graduated pipette to measure the sample of wine instead of a 10cm³
pipette followed by a 5cm³ pipette.

39
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