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Downloaded from (by Dr Richard Walding)

The goal of this analysis is to determine the amount of ascorbic (vitamin C) in a sample of
fruit juice.

Preparing Solutions

1% Starch Indicator Solution

1. Add approximately 0.5 g soluble starch to 50 mL near-boiling deionised water.
2. Mix well and allow to cool before use.

Iodine Solution (approximately 0.002 M potassium iodate)

Note: This can be called the iodate or the iodine solution as the iodate ion is converted to
iodine when it is made up.
1. Dissolve 5.00 g potassium iodide (KI) and 0.268 g potassium iodate (KIO3) in 200 mL
deionised/distilled water.
2. Add 30 mL of 3 M sulfuric acid.
3. Pour this solution into a 500 mL graduated cylinder and dilute it to a final volume of 500
mL with deionised water.
4. Mix the solution.
5. Transfer the solution to a 500 mL plastic storage bottle. Label it as iodine solution.

Vitamin C Standard Solution

1. Weigh out accurately about 0.250 g vitamin C (ascorbic acid) into a 250 mL volumetric
flask. Add about 100 mL distilled water and swirl to dissolve.
2. Add distilled water to make up to the mark. Label the flask as your vitamin C standard

Standardizing the Iodine Solutions

1. Using a pipette, add 25.00 mL aliquot of vitamin C standard solution to a 125 mL conical
flask. Note: this would contain 0.0250 g of vitamin C.
2. Add 10 drops of 1% starch solution.
3. Rinse your burette with a small volume of the iodine solution and then fill it. Record the
initial volume.
4. Titrate the solution until the endpoint is reached. This will be when you see the first
sign of blue color that persists after 20 seconds of swirling the solution.
5. Record the final volume of iodine solution. The volume that was required is the starting
volume minus the final volume.
6. Repeat the titration at least twice more. The results should agree within 0.1 mL.

Titrating Juice Samples

You titrate samples exactly the same as you did your standard. Record the initial and final
volume of iodine solution required to produce the color change at the endpoint.
1. Add 25.00 mL of juice sample to a 250 mL conical flask.
2. Titrate against iodine solution in the burette until the endpoint is reached. It is when you
get a color that persists longer than 20 seconds. See photos below:

25 mL orange juice and the At the end point Way past the end point
first addition of the iodine

3. Repeat the titration until you have at least three titres that agree to within 0.1 mL.
Note: if the titres are too big (>25 mL) consider cutting back the aliquot (sample) of
orange juice to 10 mL or less.

Titration Calculations
Approach Number 1: The first uses a simple ratio approach:
1. Calculate the mL of titrant (iodine) used for each flask of the vitC standard solution.
Take the titres you obtained and average them.
2. Calculate how much vitamin C was in the 25 mL sample of standard vitC solution: 250
mL volumetric flask contains 0.250 g vitC, so 25 mL contains 0.0250 g vitC.
3. Determine how much vitamin C was in the 25 mL sample of juice: For example, if you
needed an average of 10.00 mL of iodine solution to react with the 0.0250 grams of
vitamin C, but only needed 6.00 mL of iodine solution to react with your 25 mL of juice,
the mass of vitC in your juice is:
6/10 x 0.0250 g Vit C = 0.015 g Vit C in that 25 mL sample of juice.
3. Keep in mind the volume of your sample, so you can make other calculations, such as
grams per liter. For a 25 mL juice sample, for example:
0.015 g / 25 ml = 0.015 g / 0.025 L = 0.60 g/L of vitamin C in that sample

Approach Number 2: The traditional stoichiometry and balanced equations. For a top
mark in a Senior Chemistry EEI I would use this set of calculations as it shows the reactions
taking place.

This second method is by iodine titration and uses cheaper and more easily obtained
chemicals. In essence, you make up a potassium iodide (KIO3)and potassium iodate (KI)
solution in acid. The iodate ion IO3- reacts with the iodide ion I- in acidic H+ solution to form
iodine I2 and water H2O. The I2 reacts with the ascorbic acid solution.
2 IO3- +10 I- + 12 H+ 6 I2 + 6 H2O

During the titration the iodine reacts with ascorbic acid C6H8O6 in your sample to form
dehydroascorbic acid C6H6O6, iodide ions I- and hydrogen ions H+

6 I2 + 6 C6H8O6 6 C6H6O6 +12 I- + 12 H+

When all of the ascorbic acid is used up by the iodine at the end point, the excess iodine
turns the starch blue.

The overall reaction can be seen by the addition of the above reactions and the coefficients

IO3- + 5 I- + 3 C6H8O6 6 I- + 3 C6H6O6 + 3 H2O

The stoichiometric ratio is:

1 IO3- + 3 C6H8O6 or n (IO3-)/1 = n (C6H8O6)/3

C iodate x V iodate = (m vitc/ M vitc)/3

Sample calculations for Method Number 2:

Standard vitamin C solution

Mass vitC = 0.260 g, volume of solution = 250.0 mL (0.250 L), M (vitC) = 176.12 g/mol
C = n/V = m/M/V = 0.260/176.12/0.250 = 0.005905 M

A 25.00 mL aliquot of 0.005905 M vitC solution required a 20.25mL titre of iodate (iodine)
C iodate x V iodate = C vitC x V vitC/3
C iodate = (C vitC x V vitC/ V iodate)/3 = (0.005905 x 0.0250/0.02025)/3 = 0.002430 M

Titration of orange juice

A 25.00 mL aliquot of orange juice required a titre of 8.65 mL of 0.002430 M iodate solution.

n iodate = n vitC/3
C iodate x V iodate = (m vitC/M vitC)/3
m VitC = C iodate x V iodate x M vitC x 3
m vitC = 0.002430 x 0.00865 x 176.12 x 3 = 0.011106 g (in 25 mL of orange juice).
m VitC in 100 mL juice = 0.011106/25.00 x 100.0 = 0.04442 g/100 mL = 44.42 mg/100 mL.
This could be expressed to one decimal place as 44.4 mg/100 mL.

Note: orange juice usually ranges from about 50-80 mg/100 mL when fresh .