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Pre Lab Questions for Week 1 "Chemistry of White Wine: Part A"

Name:__________________________________

You should have four questions on this pre-lab assignment. Always read over the experiment and
complete all pre-lab questions prior to coming to your laboratory session.

Next week print out your pre-lab questions using the Pre-Lab link on the laboratory web page. Your
drawer number is used for both your ID# as well as your password.

Question:1
What chemical component(s) of wine will you measure in part A of this lab?

Question:2
Write the Lewis electron-dot formula for ethanol. Circle the portion (functional group) of the molecule
that makes ethanol an alcohol. (If necessary, use your glossary to look up ethanol and/or functional
groups.)

Question:3
A sample of wine was diluted 1:10 and analyzed for alcohol content. A calibration curve was created
from a set of standard solutions producing a plot of absorbance vs. percent alcohol. The calibration curve
had a slope of 8.05 x 10-2 with an intercept of 6.01 x 10-3. Keeping in mind that the wine sample was
diluted by a factor of ten, what is the percent, by volume, of alcohol in your sample if the absorbance of
your sample was 0.652?

Question:4
The density of pure ethanol (alcohol) is 0.789 g/mL. The percent, by volume, of alcohol in a sample of
white wine is 1.3%.
What is the density of the wine sample (use the table given in the experiment)?

What is the percent, by mass, of alcohol in the sample?

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CHEMISTRY OF WHITE WINE

Adapted with permission from the University of Illinois; Revised 2013.

INTRODUCTION

The fermentation of fruit juice producing alcoholic beverages is one of the oldest ways in which man has
used chemistry to serve himself. Today, the manufacture of wines is a multi-million dollar industry in
which chemistry plays a central role.

The process of wine making begins with growing and harvesting the fruit (usually grapes). Next the
juice, called must, is extracted from the grapes. The must is the reagent for the fermentation process.
Fermentation is an oxidation carried out in an anaerobic (no oxygen) environment by microorganisms that
grow and thrive by breaking down sugars. Yeasts are the most common microorganisms used to carry out
the fermentation process, but there are many other species, primarily bacteria, which can also cause
fermentation.

The end products of yeast fermentation are carbon dioxide and ethyl alcohol (ethanol):

Yeast + C6H12O6 → 2 C2H5OH + 2 CO2

where C6H12O6 is the molecular formula for a sugar, fructose, and its many isomers, including glucose
and galactose.

The wine making process has been refined over the centuries. There are many schools of thought on the
proper length of time to ferment the wine, the temperature at which the fermentation should be carried
out, the length of time the wine should age, the kind of container in which the wine should age, the
position in which it should age, the final alcohol content, the final sugar content, etc.

In addition to general properties, certain wines have special characteristics. Champagne and other
“sparkling” or bubbly wines undergo a second fermentation after being bottled. The CO2 given off in this
second fermentation is unable to escape and is preserved as “sparkle”. Some wines, notably the ports,
sherries, and Madeiras, have higher alcohol contents than the average table wine because brandy or some
other distilled spirit with a higher alcohol percentage has been added. Some particularly sweet dessert
wines are made from grapes that have been permitted to rot while still on the vine. The grapes are
attacked by fungus, Botrytis cinerea, which drastically reduces the liquid content of the grape but leaves
the amount of sugar unchanged.
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Investigation of the Components of Wine by Chemical Methods

Commercial wine making today is a scientific process. All the steps of fermentation are rigidly
controlled, and the final product is subject to numerous regulations as to alcohol content, sugar content,
acidity, percent of each kind of grape which can be used in its production, amount of preservatives, etc.

In this experiment, you will analyze three components of wine: alcohol content, acidity, and sufur
dioxide content.

PART A. Determination of Alcohol Content

Ethanol and certain other alcohols react with ammonium hexanitrato cerium(IV),
(NH4)2Ce(NO3)6, to yield a corresponding carboxylic acid and a cerium(III) complex. The
reaction forms a deep red intermediary complex as the alcohol is oxidized. This colored species
obeys Beer’s Law, meaning that the absorbance of light by the colored substance is directly
proportional to the concentration of the substance in solution:

A = εlc

where A = absorbance
ε = molar absorptivity
l = path length of cell
c = concentration

A Beer’s Law plot of absorbance vs. concentration of the red complex, called a calibration curve,
allows for the determination of the quantity of alcohol oxidized by ammonium hexanitrato
cerium(IV). By measuring the absorbance of the complex formed, the percent alcohol in a
sample can be determined from the calibration curve.

PART B. Acid Content Analysis

In order to determine the acidity of the wine, you will need to perform a titration. The acidity in
wine is primarily due to tartaric acid, a dicarboxylic acid with the chemical formula:

HOOCCHOHCHOHCOOH

Tartaric acid, H2C4H4O6, is a weak diprotic acid with two acidic protons; these two acidic protons
have been highlighted in the chemical formula above. Because tartaric acid has two acidic
protons, neutralization of each mole of this acid will require two moles of NaOH base:

H2C4H4O6 + 2 NaOH → Na2C4H4O6 + 2 H2O

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PART C. Sulfur Dioxide Analysis

The sulfur dioxide concentration will also be determined by titration; however the reaction is a
little different from what you have seen before. Keep in mind that the technique is the same, a
titration, and the quantitative information obtained in this case is similar.

Sulfur dioxide, SO2, is used commercially as a preservative in wines to protect the wine from
bacteriological growth and subsequent infection. Sulfur dioxide is generated by the addition of
potassium metabisulfite to the slightly acidic must:

K2S2O5 + H2O → 2 KHSO3

KHSO3 + H3O+ → SO2 + 2 H2O + K+

The preservative’s concentration is strictly regulated because the added sulfites and the SO2
produced from the reaction can be harmful to certain people. Legal limits vary from country to
country, from 200-450 mg/L. Much of the added SO2 combines with other components of wine
leaving about 20-40 mg/L free in solution to protect the product from spoilage.

In the laboratory, the concentration of free SO2 can be determined by the following reaction:

SO2(aq) + I2(aq) + 2H2O(1) → 2 I-(aq) + SO42-(aq) + 4 H+(aq)

An aqueous solution of iodine is reddish-brown, whereas the products of the reaction are
colorless. As iodine is added, the color of the wine will remain unchanged until all the sulfur
dioxide is consumed. As soon as iodine is present in excess, the following reaction occurs:

I2 + I- → I3-

The triiodide ion is slightly yellow. However, if starch is added, a dark blue-black starch-
triiodide complex forms and serves as an endpoint indicator:

I3- + starch → blue complex

To measure the total concentration of SO2, sodium hydroxide is added to the wine to break down
bisulfite complexes. Then it is acidified and titrated using the same technique.
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LABORATORY PROCEDURE

“White” wine samples are provided, and some reagents are set out in pump dispensers. Analyze the
brand of wine assigned by your instructor.

PART A. Measuring the Alcohol (ethanol) Content of Wine

1. Double click the “Logger Pro” icon and allow the screen to open.

2. The Spectrometer needs to be powered for about 5 minutes before using so do this step before
preparing your solutions. Do not use the Go!Link with the spectrometer. Plug the Spectrometer
via provided USB cable to the computer USB port.

3. Prepare Dilutions of the Stock Ethanol Solution:

a. Prepare and load a 25 mL buret to deliver the volumes of ethanol stock solution (10% by
volume, % v/v) shown in the table below. Cap the buret to reduce evaporation of ethanol.

b. Load a 50 mL buret with deionized (DI) H2O.

c. Clean and dry eight labeled large test tubes (TT). Deliver the volumes given below into the
TT. Mix well.

TT Stock Ethanol soln DI H20 Final Ethanol

(mL) (mL) concentration
(%v/v)
A1 0.0 10.0 0
B1 1.0 9.0 1.0
C1 2.0 8.0 2.0
D1 3.0 7.0 3.0
E1 4.0 6.0 4.0
F1 5.0 5.0 5.0
G1 6.0 4.0 6.0
U1 1.0 mL wine 9.0 Unknown

4. Prepare Standard Ethanol Solutions

a. Clean and dry a second set of eight large test tubes (TT) and label them A2, B2, ….

b. Rinse the pipet with about 1.0 mL of the contents of TT A1 and discard. Carefully, pipet 1.0
mL of the contents of TT A1 into TT A2. Repeat this process for the rest of the solutions.

c. Add 5.0 mL of color reagent from the pre-set pump dispenser to each TT in this second set.
Mix well. Fold a sheet of paper over the TT to protect the contents. The contents of TT A2
will now be used as the blank in the calibration of the spectrometer as well as in the
generation of the calibration curve. The contents of TT B2 will be the most dilute solution.
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d. Let the color develop for 5 minutes then immediately begin making absorbance readings.
Read over steps 5 and 6 during this 5 minute wait. The solutions in this second set of test
tubes are the solutions that will be used in the remainder of this experiment.

5. Calibrate the Spectrometer ; Do not unplug the spectrometer during this experiment or you will
have to start over.

a. Return to the Logger Pro screen on the computer. Click: Experiment ; Calibrate ;
Spectrometer:1.

b. Allow the lamp to warm up for 90 seconds as displayed on the computer screen.

c. Only touch the ridged faces of the cuvette, never touch the clear faces. Rinse and fill one
cuvette (about ¾ full) with the blank (TT A2). Rinse and fill a second cuvette with the
most dilute solution (TT B2). Gently blot (don’t scratch the sides of the cuvette) any
drips on the outside of the cuvette with a Kimwipe. These cuvettes will be used again in
the next few steps so keep them handy and keep track of what the contents are in each.

d. Place the cuvette containing the blank in the spectrometer so that one of the clear sides is
aligned with the white arrow at the top of the cuvette slot. Click: “Finish Calibration” ;
OK. Remove the cuvette from the spectrometer but keep it handy.

6. Determine the Wavelength of Maximum Absorbance

a. Place the cuvette containing the most dilute solution in the spectrometer.
Click: Experiment ; Data Collection ; Full Spectrum ; Done. Click the rainbow icon
labeled “Absorbance=…” in the upper left hand corner of the window. Change the
“Wavelength Range” to 380-750nm. Close this box by clicking the “x” in the upper right
hand corner of the window.

b. Click the small green triangle in the toolbar labeled “Collect”. After the line graph
appears on the screen, click the small red square in the toolbar labeled “Stop”.

c. To automatically store the maximum wavelength go to the toolbar and select:

Experiment ; Store Latest Run.

7. Generate the Calibration Curve

a. In the toolbar click: Experiment ; Data Collection ; choose Events with Entry in the
Mode box. Highlight the word Event in the Column Name box and replace it with
Concentration. Put percent in the Units box. Clear “Short Name”. OK. You are ready
to begin collecting data. Remove the cuvette from the spectrometer but keep it handy.

b. Place the cuvette containing the blank back in the spectrometer. Click the begin data
collection button (triangle) in the toolbar labeled “Collect”. When the absorbance
reading stabilizes, click the KEEP button located in the toolbar just to the right of the Red
Stop Button. Type in the concentration of the solution that is in the cuvette (don’t
include units). OK.
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c. Working in order of most dilute (you already have a cuvette containing the most dilute
solution) to most concentrated of the standard solutions (not the unknown), rinse and then
fill a cuvette with the solution that will be analyzed. Place the cuvette in the
spectrometer. When the absorbance reading stabilizes, click the KEEP button (be careful
that you don’t accidentally click the stop data collection button (square)), and enter the
concentration of the analyzed solution. Repeat until the absorbance of each solution has
been determined.

Pour your samples back into the appropriate TT (second set) after you have measured
each absorbance and discard them in the waste container only after you have
acceptable data.

d. When the absorbance of all standard solutions (not the unknown) has been measured,
click the stop data collection button (square) located in the toolbar.

e. To determine the equation of the line for your calibration curve click: Analyze ; Linear
Fit. A box should appear with the equation and a correlation.

To receive full credit for this lab your calibration curve must be a good, straight-line
graph, with a correlation coefficient of 99% or better (Corr: on the screen reads
0.9900 or greater). You should repeat the experiment until you get this proficiency.

Work carefully. If you need to repeat the experiment, you still must be done with the
write up and post-lab questions before the end of the lab period.

8. Each lab partner’s report must have a Logger Pro generated printout of the calibration curve
attached to it. The printout must show the graph, the information needed to generate the equation
(slope and intercept) for the line, and the correlation reading. To do this click: File ; Print.
Uncheck the “Print Visible Spectrum on Wavelength Graphs” and change the “orientation” to
landscape under properties. Be sure that the names of all lab partners are entered in the “Name”
section and that the date box is checked.

9. Rinse and fill a cuvette with your assigned wine sample from Step 4 (TT U2). Place the cuvette
in the spectrometer. Record the absorbance in your lab report once the reading has stabilized.

10. When you are done, return your bin to Lab Services.
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PART B. Determining the Acid Content of Wine

1. Clean a 50-mL buret and rinse it twice with the standard (0.05 M) NaOH solution. Load the
buret with the same NaOH solution and prepare to titrate.

2. Pipet 10.0 mL of the white wine into a conical flask. Add about 40 mL of DI water, and 3-4
drops of phenolphthalein. Titrate the sample. The endpoint is characterized by a persistent faint
pink color.

3. Repeat the titration with a second sample of wine. Adjust the sample size if necessary. Repeat
the titration a third time.

4. Measure the pH of your wine sample by adding a drop of wine to a piece of pH indicator paper.
Record this value in your laboratory report.

PART C. Determining the available SO2 concentration (C.1) and the total SO2 concentration (C.2).

C.1 Determining the Concentration of available (free) SO2

Pipet 20.0 mL (2 x 10.0 mL) of wine into a conical flask. Add 4 mL of 6 M H2SO4 and 3 mL
starch indicator. Perform a quick titration with standard iodine solution to determine the
approximate volume of I2 needed to oxidize the SO2. It may be necessary to titrate more or
less wine to get a suitable titer of iodine solution.

The end of this titration is the appearance of a dark blue color that persists for 2
minutes. Oxidation by air will then occur and the color will fade and disappear. DO NOT
ADD MORE IODINE. Titrate two more samples, or until you get consistent data. Adjust the
sample size if necessary.

C.2 Determining the Concentration of total (free and combined) SO2

Measure 10.0 mL of wine into a conical flask. Add 7.5 mL of 1 M NaOH. Swirl the flask
and wait 15 minutes. Add 4 mL of 6 M H2SO4 and 3 mL starch indicator. Titrate to the same
end point as in part C.1 and collect consistent data. Adjust the sample size if necessary.
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Specific Gravity (use as density) of Aqueous Solution Ethanol at 15ºC

Specify % Specify % Specify % Specify %

Gravity Alcohol Gravity Alcohol Gravity Alcohol Gravity Alcohol
by by by by
Volume Volume Volume Volume
1.00000 0.00 0.99417 4.00 0.98897 8.00 0.98435 12.00
0.99984 0.10 0.99403 4.10 0.98885 8.10 0.98424 12.10
0.99968 0.20 0.99390 4.20 0.98873 8.20 0.98413 12.20
0.99953 0.30 0.99376 4.30 0.98861 8.30 0.98402 12.30
0.99937 0.40 0.99363 4.40 0.98849 8.40 0.98391 12.40
0.99923 0.50 0.99349 4.50 0.98837 8.50 0.98381 12.50
0.99907 0.60 0.99335 4.60 0.98825 8.60 0.98370 12.60
0.99892 0.70 0.99322 4.70 0.98813 8.70 0.98359 12.70
0.99877 0.80 0.99308 4.80 0.98801 8.80 0.98348 12.80
0.99861 0.90 0.99295 4.90 0.98789 8.90 0.98337 12.90
0.99849 1.00 0.99281 5.00 0.98777 9.00 0.98326 13.00
0.99834 1.10 0.99268 5.10 0.98765 9.10 0.98315 13.10
0.99819 1.20 0.99255 5.20 0.98754 9.20 0.98305 13.20
0.99805 1.30 0.99241 5.30 0.98742 9.30 0.98294 13.30
0.99790 1.40 0.99228 5.40 0.98730 9.40 0.98283 13.40
0.99775 1.50 0.99215 5.50 0.98719 9.50 0.98273 13.50
0.99760 1.60 0.99202 5.60 0.98707 9.60 0.98262 13.60
0.99745 1.70 0.99189 5.70 0.98695 9.70 0.98251 13.70
0.99731 1.80 0.99175 5.80 0.98683 9.80 0.98240 13.80
0.99716 1.90 0.99162 5.90 0.98672 9.90 0.98230 13.90
0.99701 2.00 0.99149 6.00 0.98660 10.00 0.98219 14.00
0.99687 2.10 0.99136 6.10 0.98649 10.10 0.98209 14.10
0.99672 2.20 0.99123 6.20 0.98637 10.20 0.98198 14.20
0.99658 2.30 0.99111 6.30 0.98626 10.30 0.98188 14.30
0.99643 2.40 0.99098 6.40 0.98614 10.40 0.98177 14.40
0.99629 2.50 0.99085 6.50 0.98603 10.50 0.98167 14.50
0.99615 2.60 0.99072 6.60 0.98592 10.60 0.98156 14.60
0.99600 2.70 0.99059 6.70 0.98580 10.70 0.98146 14.70
0.99586 2.80 0.99047 6.80 0.98569 10.80 0.98135 14.80
0.99571 2.90 0.99034 6.90 0.98557 10.90 0.98125 14.90
0.99557 3.00 0.99021 7.00 0.98546 11.00 0.98114 15.00
0.99543 3.10 0.99009 7.10 0.98535 11.10
0.99529 3.20 0.98996 7.20 0.98524 11.20
0.99515 3.30 0.98984 7.30 0.98513 11.30
0.99501 3.40 0.98971 7.40 0.98502 11.40
0.99487 3.50 0.98959 7.50 0.98491 11.50
0.99473 3.60 0.98947 7.60 0.98479 11.60
0.99459 3.70 0.98934 7.70 0.98468 11.70
0.99445 3.80 0.98922 7.80 0.98457 11.80
0.99431 3.90 0.98909 7.90 0.98446 11.90
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DATA AND ANALYSIS SHEET: CHEMISTRY OF WHITE WINE: WEEK 1

Name: ________________________________________
Date _____________ Lab Partner ___________________________________

Part A: (Be sure to attach a copy of the calibration curve.)

Equation of line for the calibration curve: __________________________________________________

Correlation: _______________________

Assigned Wine Sample: _____________________________

Absorbance of your assigned (diluted) wine sample: ____________________________

Using the equation for your calibration curve, determine the percent, by volume, of ethanol in the
analyzed (diluted) sample.

Percent, by volume of ethanol in the analyzed (diluted) sample: ____________________

You diluted your wine sample by a factor of 10 in order to take a measurement.

Percent, by volume, of alcohol (ethanol) in your actual wine sample: ___________________

Use the table that is given to determine the density of your wine sample. When using the table, assume
that density and specific gravity are the same and that both are temperature independent.

Density of your wine sample: __________________________

The density of pure ethanol (alcohol) is 0.789 g/mL. Calculate the percent, by mass, of alcohol (ethanol)
in your wine sample. Show your work.

Percent, by mass, of alcohol (ethanol) in your wine sample: ____________________

Compare your results for alcohol content with the data from the label for the bulk wine you tested.
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DATA AND ANALYSIS SHEET: CHEMISTRY OF WHITE WINE; WEEK 2

Name: ________________________________________
Date _____________ Lab Partner ___________________________________

Assigned sample: _________________

Molar Concentration (exact) of the standard NaOH solution: _______________
Molar Concentration of the I2 solution: _____________________

PART B: Determining the Acid Content of Wine

Trial 1 Trial 2 Trial 3
Volume of wine sample
Final buret reading, NaOH
Initial buret reading, NaOH
Total volume NaOH used
Molarity of tartaric acid
Average molarity of tartaric acid

pH of your wine sample

PART C.1: Determining the Concentration of available (free) SO2

Trial 1 Trial 2 Trial 3
Volume of wine sample
Final buret reading, I2
Initial buret reading, I2
Total volume I2 used
Available SO2 concentration (M)
Average available SO2 concentration (M)
Average available SO2 concentration (ppm)

PART C.2: Determining the Concentration of total (free and combined) SO2
Trial 1 Trial 2 Trial 3
Volume of wine sample
Final buret reading, I2
Initial buret reading, I2
Total volume I2 used
Total SO2 concentration (M)
Average total SO2 concentration (M)
Average total SO2 concentration (ppm)
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DATA AND ANALYSIS SHEET: CHEMISTRY OF WHITE WINE; WEEK 2

Sample Calculation(s) (Part B):

Sample Calculation(s) (Part C.1):

Sample Calculation(s) (Part C.2):