Determination of Vitamin C in Foods

©2013 by David A. Katz. Al rights reserved.

Object
The object of this experiment is to measure the amount of ascorbic acid in common foods using an oxidation
reduction reaction.

Introduction
Vitamin C, has the chemical name ascorbic acid. It is a water soluble vitamin. Although it is important for
good health, humans do not have the ability to make their own vitamin C and must obtain it through diet or
take it in vitamin supplements. Citrus fruits, potatoes and some green vegetables are known to be good
sources of vitamin C. (See Table 1) Plants synthesize the compound for the growth, development, and
protection of the plant. The exact pathway for its synthesis is not well understood.

Amount of   Amount of
Plant source   Vitamin C Plant source   Vitamin C
(mg / 100g) (mg / 100g)  
Rose hip 426   Spinach 30 
Red pepper   190   Potato 20 
Parsley   130   Green Beans 16 
Broccoli   90   Tomato 10 
Brussels sprouts   80   Watermelon 10 
Elderberry   60   Banana 9 
Strawberry   60   Carrot 9 
Orange   50   Apple 6 
cantaloupe   40   Lettuce 4 
grapefruit   30   Raisin 2 

Table 1. Amount of Vitamin C in Various Plants

Vitamin C is notoriously easy to destroy. Orange juice may lose half of its vitamin C in a week in the
refrigerator, but cut fruit may lose much less after 6 days (Some examples: mango, strawberry, and
watermelon: less than 5%, Pineapple: 10%, Kiwi: 12%, and Cantaloupe: 25%, after 6 days). Cooking also
destroys vitamin C. The US Department of Agriculture reports the following vitamin C levels:

Amount of vitamin C, mg/100g

Vegetable Raw Frozen Frozen then boiled
Green beans 12.2 9.7 4.1
Broccoli 89.2 64.9 40.1

Note: Freezing fresh vegetables requires blanching, i.e., boiling

for a short period to stop enzyme actions in the plant.

Some cooks had a practice of adding sodium bicarbonate, NaHCO3, to vegetables to maintain a bright
green color. That practice essentially destroys the Vitamin C in the vegetables.

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Vitamin C is required for the synthesis of collagen, an important structural component of blood vessels,
tendons, ligaments and bone. It also is important in the synthesis of the neurotransmitter norepinephrine,
which is critical to brain function and can affect mood.

Vitamin C is also a highly effective antioxidant. In small amounts vitamin C can protect indispensable
molecules in the body, such as proteins, lipids (fats), carbohydrates, and nucleic acids (DNA and RNA),
from damage by free radicals and reactive oxygen species that can be generated during normal metabolism
as well as through exposure to toxins and pollutants. It has been recommended that vitamin C, taken in
large quantities on a daily basis, may prevent the common cold. Some recommended dietary allowances
for vitamin C are listed in Table 2.

Age Male Female Pregnancy Lactation

0–6 months 40 mg* 40 mg*
7–12 months 50 mg* 50 mg*
1–3 years 15 mg 15 mg
4–8 years 25 mg 25 mg
9–13 years 45 mg 45 mg
14–18 years 75 mg 65 mg 80 mg 115 mg
19+ years 90 mg 75 mg 85 mg 120 mg
Smokers Individuals who smoke require 35 mg/day more
vitamin C than nonsmokers.

Table 2. Recommended Dietary Allowances (RDAs) for Vitamin C

Ascorbic acid is the chemical name for vitamin C. As the name indicates vitamin C is an acid. Therefore, a
base such as sodium hydroxide can neutralize it. Ascorbic acid is also easily oxidized. Both of these
reactions can be used for quantitative analysis of the compound.

Figure 1. The structure of ascorbic acid, C6H8H6

(MW = 176.12 g/mol)

In this experiment, you are performing an oxidation-reduction titration. You will first determine the
concentration of the iodine solution by titrating a solution of ascorbic acid. Next, you will titrate fruit juices
or a fruit or vegetable extract. Ascorbic acid reacts with iodine through the following reaction:

C6H8O6 + I2  C6H6O6 + 2 I- + 2 H+

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The equivalence point of the titration will be determined using a starch indicator. Molecular iodine
combines with starch to make a dark blue complex. In the titration reaction, molecular iodine is added in
the form of the iodine solution. In the presence of ascorbic acid, the iodine is reduced to colorless iodide
ion which does not react with starch to make a colored complex. When all of the ascorbic acid has been
titrated, any additional iodine added, present in the solution as molecular iodine, will combine with the
starch to make the solution blue. The appearance of the blue solution will signal the equivalence point. As
with the acid-base titration, the iodine solution must be standardized against a known solution of ascorbic
acid, which you will prepare from the solid.

Safety

Goggles or safety glasses must be worn at all times in the laboratory.

The iodine solution is an irritant and will stain the skin and clothing. The solution does give off a small
amount of iodine vapors. Iodine vapors are toxic by inhalation. Work in a well ventilated area. Keep the
iodine solution covered.

Disposal
Dispose of all solutions in the waste containers provided.

Materials Needed
Ascorbic Acid
Iodine Solution
2% Starch solution (indicator)
125 ml Erlenmeyer flasks
Buret
Magnetic stirrer with stir bar
Small funnel for filling buret
Blender
Knife
Small cutting board (wood or plastic)
Cheesecloth
Funnel
Scissors
Optional: gloves

Experimental Procedure
 
Standardization of the iodine solution
You will use a solution of ascorbic acid to standardize the iodine solution.

1. Weigh three samples of solid ascorbic acid, approximately 0.05g to 0.10 g each.

2. Place each sample in a numbered 125 ml Erlenmeyer flask. Add about 30ml distilled water and 4
to 5 drops of starch solution to each flask.

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3. Rinse and fill a 25 or 50 mL buret with the iodine solution and titrate the ascorbic acid solution. Be
sure to record the initial and final volume readings from the buret.

4. After the data is collected, calculate the molarity of the iodine solution.

5. Repeat the procedure for the other two ascorbic acid solutions

Determination of Vitamin C in a Fruit Juice or Food

Each group will run two sets of titrations, one with fresh fruit juice and refrigerated juice (e.g., juice from
an orange and packaged orange juice) OR one with a fresh vegetable and one with frozen vegetable (e.g.,
broccoli and frozen broccoli).

Preparation of samples

For fresh fruit juice from oranges, grapefruits, or lemons.

If using fresh fruit, squeeze at least 100 mL of the juice from the fruit. If the juice contains pulp
and any seeds, strain 100 mL of juice through cheesecloth.

For packaged fruit juice.

If the juice contains pulp, strain 100 mL of juice through cheesecloth.

For fresh or frozen fruits and vegetables.

Cut a 100 g sample into small pieces.
 
Place the sample into a blender or food processor, add 50 mL distilled water and blend to a pulp.

Strain the fruit or vegetable pulp through cheesecloth, washing the pulp with a few 10 mL portions
of water and collecting all filtrate in a 150-mL or 250-mL beaker.
 
Pour the filtrate into a 100 mL graduated cylinder and dilute the solution to 100 mL with distilled
water.

Titration of the fruit juice or extract

 
1. Measure 20 mL of your sample solution into a 125 mL Erlenmeyer flask. Add 25 mL of distilled
water and 1 mL of starch indicator solution.
 
2. Titrate the sample with your standardized iodine solution from the first part of this experiment. The
endpoint of the titration occurs when you obtain a permanent dark blue-black color due to the starch-
iodine complex.
 
3. Repeat the titration twice more with additional 20 mL aliquots of your sample solution. Your titration
results should agree to within 0.5 mL. If they do not agree, see your instructor. You may have to do
an additional titration.

4. If you started with juice or extract from fresh fruit or vegetable, in step 1, above, repeat the series of
titrations using samples prepared from the packaged juice or extract from frozen vegetables. If you

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stared with the packaged juice and frozen vegetable, then use samples prepared from the fresh juice
or vegetable.

5. Determine the amount of ascorbic acid present in the samples of fruit juice or vegetable you titrated.
Compare your result to the vitamin C content listed on the label of the packaged juice, from Table 1
in this experiment, or from data obtained on the Internet.

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Determination of Vitamin C in Foods
Data and Results
Name_____________________________________________ Course/Section ______________________

Partner’s Name (If applicable) _________________________________________ Date _______________ 

 

Standardization of the iodine solution

Mass of Initial Volume Final Volume of Volume of Concentration of
ascorbic acid of Iodine Iodine solution, Iodine Solution Iodine Solution
used, Solution, mL mL used, mL M
g
         

         

         

         

  Formula for Miodine:

1 moleascorbic acid 1000 mL

MIodine  massascorbic acid   L
 
176.12 gascorbic acid VolumeIodine solution , mL

Sample calculation for Miodine (Show the set-up for one calculation)
 

Average concentration of iodine solution  _____________________ M 

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Determination of Vitamin C in a Fruit Juice or Food
 
Fruit or fruit juice used __________________________________________________________

Mass of fresh fruit or vegetable OR volume of fresh fruit juice used __________________________

Volume of fruit  Initial Volume of  Final Volume of  Volume of  Concentration of 

juice or extract  Iodine Solution,  Iodine solution,  Iodine Solution  Ascorbic Acid in 
used,  mL  mL  used, mL  Sample 
mL  mg 
         

         

         

         

Mass of frozen fruit or vegetable OR volume of packaged fruit juice used _______________________

Volume of fruit  Initial Volume of  Final Volume of  Volume of  Concentration of 

juice or extract  Iodine Solution,  Iodine solution,  Iodine Solution  Ascorbic Acid in 
used,  mL  mL  used, mL  Sample 
mL  mg 
         

         

         

         

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  Formula for Concentration of Ascorbic Acid in your sample:

mgascorbicacid  Miodine solution  mLiodine solution 176.12 g

mole
Sample calculation for mgascorbic acid (Show the set-up for one calculation)

Summary of titration results

Average concentration of Ascorbic Acid in fresh fruit

or vegetable samples _____________________ mg

Average Volume of fresh fruit juice or extract used for

each titration _____________________ mL

Average concentration of Ascorbic Acid in packaged

fruit or frozen vegetable samples _____________________ mg

Average Volume of packaged fruit juice or frozen

vegetable extract used for each titration _____________________ mL

Concentration of Ascorbic acid in Food

(Note: This equation gives answer in mg Ascorbic acid/100 g fruit. We are assuming that 1 mL of fruit
juice or fruit extract has a mass of 1 g.)

Average Conc.  Ascorbic acid 1 mLFruit juice

Concentration Ascorbic Acid in fruit used=   100
AverageVolumeFruit juice 1 gFruit juice

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Sample calculation for Concentrationascorbic acid (Show the set-up for one calculation)

Final Results

Average concentration of Ascorbic Acid in fresh fruit juice

or vegetable tested __________________ mg/100 g

Average concentration of Ascorbic Acid in packaged fruit

juice or frozen vegetable tested __________________ mg/100 g

Questions

1. What is the concentration of vitamin C in the juice or fruit tested according to the package label,
Table 1 (in the experiment), or from Internet data?

2. How does the concentration of vitamin C from your result of this experiment compare to the
values in question 1, above?

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3. Can you explain any differences you observed in question 2, above?

4. Your group tested fresh fruits, vegetables, or juices, and frozen or packaged fruits, vegetables, or
juices, how does their vitamin C concentrations compare?

5. Do you have any recommendations for buying, cooking and storing juices, fruits, and vegetables?

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