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A NATURAL APPROACH TO CHEMISTRY 113
13D: Determining the Amount of Vitamin C
How can we determine the amount of ascorbic acid in vitamin C tablets and in
different foods?
We will determine the amount of vitamin C using a simple titration method. The chemical name for
vitaminC is ascorbic acid. This procedure relies on a series of chemical reactions that cause an observable
color change to occur. To help you better understand what you are seeing when the color change occurs, it
is helpful to review some relevant chemical reactions.
In equation two, Vitamin C reacts with iodine (I
2
) and becomes oxidized. This is the most important
chemical reaction taking place. However, to produce the iodine (I
2
)

we must first carry out another reaction.
Adding solid potassium iodide (KI) and hydrochloric acid (HCl) and the titrant, potassium iodate (KIO
3
),
creates iodine and water according to the following reaction:
IO
3

+5I

+6H
+
→ 3I
2
+3H
2
O
As soon as the iodine (I
2
)

is created it reacts with the vitamin C, causing the vitamin C to become oxidized.
Ascorbic acid, or vitamin C, is very important for our
bodies. Vitamin C is required by our bodies to produce
collagen in our connective tissues. Vitamin C thus helps
us to maintain healthy teeth, blood vessels, and bones.
Vitamin C has also been found to function as an
antioxidant, and it aids in the prevention of cancer.
Antioxidants are molecules that can protect our cells from
being damaged. Antioxidants such as vitamin C are able
to absorb harmful reactive molecules before they can
damage our tissues. In this investigation, a titration will be
used to determine the amount of vitamin C that is
contained in “vitamin C” tablets and in different foods and
juices. The recommended daily amount of vitamin C is 60
mg.
Materials
• 250 mL beaker or Erlenmeyer flask
• Vitamin C solution (2 mg/mL),
obtained by grinding one 500 mg
tablet and dissolving in 250 mL of
distilled water
• Dropper bottle
• 10 mL graduated cylinder
• 10 g of starch
• Distilled water bottle
• 20 mL of 0.010 M KIO
3
solution
• 1.0 g KI
• 5.0 mL 1.0 M HCl
• Orange juice and grapefruit juice
• Citrus fruits and juicer
• Cheesecloth
• Medium funnel
2)
1)
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Investigation 13D: Determining the Amount of Vitamin C
114 A NATURAL APPROACH TO CHEMISTRY
After all of the vitamin C has reacted, the I
2
concentration builds up and then begins to react with I

in
solution to form the triiodide ion (I
3

). The triiodide ion complexes with the starch to give a deep bluish-
black color that we can see:
I
2
+I

→ I
3

and I
3

+starch → iodide–starch complex
deep blue color
When this color appears, we know all of the vitamin C has reacted. It is the formation of the triiodide ion
that signals the endpoint of the titration.
Once we know how much titrant (KIO
3
) is required to reach the endpoint, we can use the volume of KIO
3
to determine the moles of KIO
3
. Using stoichiometry we can use the moles of KIO
3
to find the moles of
iodine (I
2
)

and therefore the moles of vitamin C present in our experiment.
Part 1: Calibrate your dropper bottle or pipette
1. We need to accurately determine the number of drops in 1.0 mL. To accomplish this, use either a
dropper bottle or a plastic beral pipet. [In Part 2, be sure you add the titrant (KIO
3
) using the same
dropper and method].
2. To calibrate your dropper, slowly add drops of water to a small 10.0 mL graduated cylinder. Stop
adding drops when the meniscus is on the 1.0 mL mark. Record the number of drops you added.
3. Next count how many drops it takes to get to the 2.0 mL mark. Record your measurement. If the
number of drops is in close agreement, take the average. If your number of drops does not agree,
repeat Steps 2 and 3.
Part 2: Verifying the amount of vitamin C in a commercial tablet
1. Measure 25.0 mL of the vitamin C
stock solution, and add it to a 250
mL Erlenmeyer flask. Your
instructor will have prepared this
ahead of time. Record the
concentration of the stock solution
in your lab notebook.
2. Prepare a 1% starch solution.
3. Measure 1.0 g of KI solid and add
it to the Erlenmeyer flask. Swirl to dissolve.
4. Add 5.0 mL of 1.0 M HCl to the Erlenmeyer flask.
5. Add 2–3 mL of the fresh 1.0% starch solution indicator to the Erlenmeyer flask. Swirl to mix.
6. Obtain a dropper bottle of 0.010 M KIO
3
titrant solution. This dropper bottle should be the same
type as the one you used in Part 1. If your instructor has the titrant in a stock bottle, measure
about 25.0 mL of the 0.010 M KIO
3
titrant solution in a clean, dry beaker and add it to a dropper
bottle yourself.
7. Now you are ready to titrate. Add 0.010 M KIO
3
solution drop by drop to the Erlenmeyer flask.
Swirl after every few drops. Initially you will see the deep blue color appear in the area that the
drop is added. Once you begin to see a deep blue color that persists throughout the solution, add
the titrant more slowly and swirl after every drop. Continue adding titrant until you obtain a
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Investigation 13D: Determining the Amount of Vitamin C
A NATURAL APPROACH TO CHEMISTRY 115
permanent blue color that lasts about 60 s. Record the total number of drops you added. The
solution may lighten after the 60 s, but it should remain fairly deep blue. Determining the “right”
color takes practice and is somewhat objective. Do your best, and then repeat the experiment.
After two or three trials, you will feel more confident at determining your endpoint.
Part 3: How much vitamin C is there in different fruits and juices?
Here we will determine the vitamin C content of a food of your choice. You
may use citrus fruits, fresh squeezed or prepared, apple juice, or any drink
containing ascorbic acid. Real lemon juice also makes a nice sample to test,
and the amount of vitamin C is on the label.
1. To begin you may use fresh or prepared juice. You will need about
25 mL of juice to have enough vitaminC for a good measurement.
If you use freshly squeezed juice, it is helpful to strain out the pulp
with cheesecloth. An easy way to strain out the pulp is to use a
funnel lined with two or three layers of cheesecloth. Pour the
sample of juice through the cheesecloth and collect the juice.
Record the exact volume of juice you will titrate.
2. Add your juice to a 250 mL Erlenmeyer flask.
3. Titrate your sample using the same steps you used to titrate the
vitamin C tablet. Be sure to add 1.0 g of solid KI, 5.0 mL of 1.0 M
HCl and 2–3 mL of starch solution.
Note: You may not see the deep blue color because of the color of your fruit juice. For instance, it
may appear brownish instead of blue. This is fine; just be sure you look for the same color change
in each trial. That is the key.
4. Repeat your experiment two or three times to obtain an accurate sense of the reproducibility of
your results. Record your results and find the average amount of vitamin C in your sample.
Part 4: Thinking about it
1. Make a data table that records the number of drops of 0.010 M KIO
3
needed to titrate 25 mL of
the vitamin C stock solution.
2. Convert your drops to milliliters using the conversion you
determined in Part 1. Record this in your data table. Here is
an example:
3. Does it matter how much distilled water was added to your
sample? Explain.
4. Which contained more vitamin C: the tablet or the food?
How do you know?
5. Look carefully at the first equation on page 113. Copy this equation into your lab notebook.
Which substance was the reducing agent? Explain. Which substance was the oxidizing agent?
Explain.
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Investigation 13D: Determining the Amount of Vitamin C
116 A NATURAL APPROACH TO CHEMISTRY
Part 5: What did you obtain for results?
1. Use the volume of your KIO
3
titrant added in liters
and the known molarity (0.010 M) to calculate the
moles of KIO
3
required to titrate the vitamin C
sample, as shown in this example:
2. Using the second equation on page 113 and
stoichiometry, convert the moles of KIO
3
to moles of
I
2
produced. See the example calculation shown in
Step 3.
3. Calculate your moles of vitamin C, from the amount of I
2
produced in Step 2. Use the first
equation on page 113, stoichiometry, and the following example to help you with this calculation:
4. Calculate the grams of vitamin C from the moles of vitamin C you determined.
5. Convert your grams to milligrams by dividing by 1,000. This number tells you how many
milligrams of vitamin C were in your 25.0 mL sample of the vitamin C stock solution, or your
food.
6. Which food contained the highest amount of vitamin C? Explain.
7. The stock solution contained a 500 mg tablet of vitamin C and 250 mL of distilled water.
Combine your results with those of your classmates and see how close your results came to this
value. For example, if each group titrated 25.0 mL of stock solution, then the total milligrams in
10 trials would give you a total volume of 250 mL of solution, and this should give you a value
close to 500 mg of vitamin C. To estimate, you could also take your trial average and multiply the
number of milligrams by 10 to see if you are close to 500 mg.
8. Based on your result, does a 500 mg tablet really contain 500 mg of vitamin C?
9. If the recommended daily allowance of vitamin C is 60 mg, how much of your food would you
need to consume to get your daily allowance? Show your calculation.
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