Vitamin C Analysis

Please review Daniel C. Harris, Quantitative Chemical Analysis (10th Edition) Ch. 16 for in-depth
discussion of the techniques in this experiment.

By completing this experiment, you will learn how to use volumetric methods, namely titration, to
precisely perform a chemical analysis. Specifically, you will learn to perform redox titrations using
iodometric methods to indicate the endpoint.

Introduction
The history of vitamin C (Ascorbic Acid) is the history of the human disease scurvy, probably the
first human illness to be recognized as a deficiency disease. Its symptoms include exhaustion,
massive hemorrhaging of flesh and gums, general weakness and diarrhea. Resultant death was
common. Scurvy is a disease unique to guinea pigs, various primates and humans. All other animal
species have an enzyme that catalyzes the oxidation of L-gluconactone to L-ascorbic acid,
allowing them to synthesize vitamin C in amounts adequate for metabolic needs.

Figure 1. Structure of L-ascorbic acid (Vitamin C).

As early as 1536, Jacques Cartier, a French explorer, reported the miraculous curative effects of
infusions of pine bark and needles used by Native Americans. These items are now known to be
good sources of ascorbic acid. However, some 400 years were to pass before vitamin C was
isolated, characterized, and synthesized. In the late 1700's, the British Navy ordered the use of
limes on ships to prevent scurvy. This practice was for many years considered to be quackery by
the merchant marine, and the Navy sailors became known as 'Limeys.' At that time, scurvy aboard
sailing vessels was a serious problem, with often up to 50 % of the crew dying from scurvy on
long voyages.

The Recommended Daily Allowance, or RDA, for vitamin C put forward by the Food and
Nutrition Board of the National Research Council is 75-90 mg/day for adults. It is recommended
that pregnant women consume an additional 10 mg/day. Medical research shows that 10 mg/day
of vitamin C will prevent scurvy in adults. There has been much controversy over speculation that
vitamin C intake should be much higher than the RDA for the prevention of colds and flu. Linus
Pauling, winner of both a Nobel Prize in Chemistry and the Nobel Peace Prize, has argued in his

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book, “Vitamin C and the Common Cold”, that humans should be consuming between 250 mg and
10 g of vitamin C, depending on one's physiology.

Vitamin C is a six-carbon molecule, closely chemically related to glucose. It was first isolated in
1928 by the Hungarian-born scientist Szent-Gyorgi, and structurally characterized by Haworth in
1933. In 1934, Rechstein worked out a simple, inexpensive, four-step process for synthesizing
ascorbic acid from glucose. This method has been used for commercial synthesis of vitamin C.
Vitamin C occurs naturally primarily in fresh fruits and vegetables. A table of some typical vitamin
C contents of foodstuffs is given below. Imagine how many lemons one would have to eat in order
to ingest 1 g of vitamin C!
Table 1. Vitamin C content in some foodstuffs.

Vitamin C (mg/100 g) Foods

100 -350 Chili peppers, sweet peppers, parsley, turnip

greens

25-100 Citrus juices, tomato juice, mustard greens, spinach,

Brussel sprouts

Green beans and peas, sweet com, asparagus,

10 -25 pineapple, cranberries, cucumbers, lettuce
< 10 Eggs, milk, carrots, beets, cooked meat

Redox Titration with Iodine

Given the widespread importance of vitamin C, many methods for analyzing it in complicated
environments have been developed. The redox method of analysis used in this lab takes
advantage of the ease of oxidation of vitamin C. As shown below, vitamin C is readily oxidized in
acidic conditions. We will use iodine (I2 that will be generated in situ) as the oxidizing agent
(electron acceptor).

Figure 2. Oxidation of Vitamin C

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When the ascorbic acid is completely reacted away, the I 2 will begin to build up. The solution will
also contain iodide ions (I-) and starch, so that the excess iodine will generate the dark colored
triiodide (I3–)/starch complex, which will signal the "endpoint" of the titration. Because it is not
practical to prepare an aqueous I2 solution for the titration, we will prepare it in situ (in the reaction
flask). To accomplish this, we will employ dropwise addition of water-soluble, colorless potassium
iodate (KIO3), which reacts with I- under acidic conditions to form I2 as shown in the reaction
below. This reaction will rapidly generate I2 in the solution to be analyzed, and this will oxidize
the vitamin C.

IO-3 (𝑎𝑞) + 6 H+ (𝑎𝑞) + 5 I- (𝑎𝑞) ⇌ 3 I2 (𝑔) + 3 H2 O (𝑙) (1)

Note that although you will be titrating with an IO 3- solution, it is the iodine produced by this
reaction that actually oxidizes the vitamin C. Using this method, we will be able to
quantitatively determine the amount of vitamin C in a sample.

Remember that we are generating the I2 in situ to use as an oxidation agent. What then, is the
reduction half-reaction?

References

[1] J. L. Roberts, J. L. Hollenberg and J. M. Postma, General Chemistry in the Laboratory, 3rd
Ed. ed., W. H. Freeman and Company, 1990.

[2] D. C. Harris and C. A. Lucy, "Ch. 16 Redox Titrations," in Quantitative Chemical Analysis,
10th ed., New York, New York: Macmillan Learning, 2020, pp. 381-402.

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 Prelab Questions
1. Write the balanced chemical equation for the titration of Vitamin C with iodine solution at
acidic pH. (Hint: You need to know that half reactions for the oxidation of ascorbic acid and
the reduction of iodide)

2. In qualitative terms, describe what is happening when a solution of KIO3 is added to a solution
containing vitamin C, KI, and starch at a pH lower than 7.

3. Briefly describe where any waste should be disposed of. Be specific, you need to be able to
identify which trash can each item of waste goes in!

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 Materials and Methods
Work in groups of 2

Purpose
In this experiment, you will employ a redox titration method to determine the percentage of
Vitamin C in both commercial tablets (with a nominal dose of 100 mg per tablet) and in an
unknown sample.

Materials
Equipment

1× Buret (50 mL) 3× Erlenmeyer flask (250 mL)

Mortar and pestle Graduated Cylinder (50 or 100 mL)
Analytical balance Graduated Cylinder (10 mL)

Chemicals

Potassium iodate, 0.01 M KIO3 CAS 7758-05-6 150 mL

While considered relatively non-hazardous in solution at this
concentration, potassium iodate is a strong oxidizer: contact with
Hazards/Safety
skin should be avoided and spills should be cleaned up
immediately.

Potassium iodide KI CAS 7681-11-0 6g

Danger! Causes mild eye, skin, respiratory tract and gastrointestinal tract
irritation. May be harmful if absorbed through the skin, ingested, or
inhaled. Chronic exposure may cause reproductive and fetal effects,
and thyroid issues. Target organs: thyroid.

Hydrochloric acid, 1 M HCl CAS 7647-01-0 30 mL

Vapor or mist may cause irritation and severe burns to the skin,
Danger! eyes, and respiratory system and may be fatal if inhaled. Contact
with liquid is corrosive to the skin and eyes. Ingestion causes
digestive track burns and potentially permanent tissue damage or
death. Target organs: Respiratory system, gastrointestinal system,
teeth, eyes, and skin.

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 Starch indicator, 1% w/v
Soluble Starch (1%) (C6H10O5)_n CAS 9005-84-9
Salicylic Acid (0.1%) C7H6O3 CAS 69-72-7 6 mL
Warning!
May cause eye, skin, gastrointestinal and respiratory tract irritation.

Vitamin C tablets and

unknown mixtures C6H8O6 CAS 50-81-7 2g
May cause skin or eye irritation; may cause respiratory tract
Hazards/Safety
irritation if inhaled.

Procedure
Analysis of Vitamin C Tablets
1. Rinse and fill a clean buret with ~0.01000 M KIO3 solution. Record the exact concentration
of KIO3 from the chemical bottle.

2. Measure the mass of a 100 mg vitamin C tablet on an analytical balance. Is the mass of the
tablet what you expected?

3. Grind the tablet using a clean, dry mortar and pestle. Weigh
out approximately one third of the powder, about 0.3 g,
using the same analytical balance you used previously.
(Save the other two-thirds for the next two trials.) Record
the mass to the nearest 0.1 mg.

4. Calculate the percent mass of vitamin C in the tablet (Use

the actual mass of the tablet and the listed dose of vitamin
Figure 3. A mortar and pestle
C). What is the predicted mass of vitamin C in your sample?
How many moles of vitamin C are expected in to be in your
sample?

5. Quantitatively transfer the weighed powder to a 250 mL Erlenmeyer flask. You will add about
50 mL of distilled water to this sample. Use some of that distilled water to rinse any remaining
powder from the weighing vessel into the flask. Stir the solution for a few minutes. Ascorbic
acid is very soluble in water. Any material left suspended in the water after stirring is not
likely to be ascorbic acid. What could this material be?

6. Add 1 g of solid KI to the flask and swirl until it dissolves.

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7. Add 5 mL of 1 M HCl and swirl the flask to mix the reagent.

8. Add 1 mL of 1 % starch indicator solution to the flask. (Note: The starch can settle out of
solution. Give it a good shake before dispensing.) Swirl the flask to thoroughly mix the
reagents.

9. Titrate the ascorbic acid solution with the KIO 3 solution. Did you remember to record the
exact concentration of KIO3? Record the initial volume reading on the buret. When using your
50 mL burets you should record all volumes to the nearest 0.01 mL. As the KIO3 solution is
added you may see a yellow or purple-blue color start to form as the endpoint is approached.
While adding the KIO3, swirl the flask to ensure reactants to mix throughout. Before the
endpoint any color that appears will vanish as the flask is swirled. The endpoint occurs when
adding a small amount of KIO3 solution (a drop or less) causes the purple-blue color to persist
throughout the flask, even after 20 seconds of swirling (Figure 4). When you have observed
this color change stop adding KIO3 and record the final volume on the buret.

Figure 4. Triiodide/startch complex endpoint. a) Before the endpoint the solution will be mostly clear. It may
appear yellow or purple-blue momentarily as you near the endpoint, but this color will rapidly dissipate upon
swirling. b) At the endpoint addition of only a drop or less will turn the entire solution purple-blue and it will
remain purple-blue through mixing.

10. Perform two more titrations using the remaining powdered vitamin C tablet as described
above. If you run out of sample before completing 3 accurate titrations request another 100
mg tablet and prepare it as before.

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Analysis of Vitamin C Unknown Sample
Obtain an unknown sample from the ISF stockroom window and record its number in your lab
notebook. This unknown is comprised of vitamin C, but with unreactive filler. It is already in a
powdered form so you will not need to use the mortar and pestle. Use no more than 0.15 g of the
sample per titration. Your job is to find the percent composition of vitamin C in your unknown
sample using the method described above. If your sample requires more than 50 mL of 0.01000 M
KIO3 to titrate, you may need to use a smaller sample mass. Dispose of any unused unknown along
with your unknown vial in the purple-blue chemically contaminated waste bin.

Cleanup and Waste Disposal

1. Rinse your buret thoroughly with distilled water and allow to drip dry. Leave the mostly dry
buret in the buret rack on the lab bench.

2. All titration solution waste should be disposed of in the “All Solutions” waste container in the
hood. Be sure to log your waste – you should know exactly how much of each component you
have added from the procedure.

3. Unused vitamin C solid waste, unknown samples and vials, and plastic droppers go in the
purple-blue chemically contaminated waste bins.

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 Postlab Questions
For all the questions that require calculation, you must show your work for at least one set of data.
You may present your work in whatever way you are comfortable (pen and paper, excel, Matlab,
etc.), but your work must be your own.

1. Report the mass (mg) of vitamin C in the tablet per mg of sample that you determined from
your titrations with proper 95% confidence intervals. Use a t-test to decide if your results
agree with the specified composition of the tablet.

2. Report the mass (mg) of vitamin C in your unknown per mg of sample with 95% confidence
intervals.

3. The oxidation of Vitamin C is actually even more facile under basic conditions. Why do you
think this is so?

4. Write an abstract for this experiment. An abstract is a brief (one paragraph or less) description
of the goal and results of the experiment.

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