Experiment 6

Determination of Ascorbic Acid Using Iodometric Titration

Method
Objectives

Part I: To determine the ascorbic acid content of vitamin C tablets using an iodometric
titration.

Part II: To determine the concentration of vitamin C in juices and real-lemon using an
iodometric titration.

Introduction

Although most mammals can synthesize vitamin C or ascorbic acid (C 6H8O6) from sugars,
man must ingest considerable quantities of ascorbic acid. The National Academy of Sciences
recommends the consumption of 60 mg of ascorbic acid per day. Vitamin C deficiency,
which typically causes abnormalities in bones and teeth, was first characterized in sailors in
the eighteenth century. These abnormalities were eliminated by compelling sailors to eat
limes, a source of vitamin C. Many vegetables also contain large quantities of vitamin C, but
ascorbic acid is commonly destroyed by many cooking processes, and hence citrus fruits are
regarded as the most reliable source of vitamin C.

Vitamin C can be determined in food by the use of an oxidation-reduction reaction.

Ascorbic acid is a mild reducing agent that reacts rapidly and quantitatively with iodine to
reduce it to iodide ion. In this experiment, a known excess of iodine is formed by the reaction
of an accurately weighed amount of iodate ion in the presence of excess iodide ion.

IO3- + 8 I- + 6 H+ 3 I3- + 3 H2O

The solubility of firstly formed iodine (I2) [IO3- + 5 I- + 6 H+ → 3 I2 + 3 H2O] is increased by

complexation with excess iodide to form triiodide ion (I3-).

I2 + I- I3- (I2I-)

Triiodide then oxidizes vitamin C to dehydroascorbic acid:

C6H8O6 + I3- + H2O C6H6O6 + 3 I- + 2 H+

ascorbic dehydroascorbic

acid acid
Once the ascorbic acid has quantitatively reacted with the iodine (as triiodide), the remaining
iodine (triiodide) is back-titrated with thiosulfate. The titrant, thiosulfate, is standardized
against iodine using the same iodate and iodide reaction.

I3 - + 2 S2O3 2- 3 I- + S4O62-

To indicate the end point, the disappearance of a blue starch colour is used. It is believed that
the blue starch color is caused by the formation of a surface, coloured complex between
iodine (as triiodide) and the ß-amylose molecule found in the starch.

Part I: Determination of ascorbic acid content in vitamin C tablets

Materials and Method

Glassware and Apparatus

 Pipettes; 10 and 50 mL
 Burette; 50 mL and stand
 Conical (Erlenmeyer) flasks; 250 mL
 Measuring (graduated) cylinders; 10 mL and 100 mL
 Glass rod

Chemicals
 Sodium thiosulfate (Na2S2O3.5H2O)
 Sodium carbonate (Na2CO3)
 Potassium iodide (KI)
 Potassium iodate (KIO3)
 Vitamin C tablets
 Starch indicator (containing HgI2 as the preservative)
 Sulfuric acid; 0.30 and 0.50 M

Procedure

A. Reagents preparation

Sodium thiosulfate solution (0.10 mol L ): A 0.100 M sodium thiosulfate solution has been
−1

prepared by dissolving about 12.40 g of solid Na2S2O3.5H2O to 500 mL mark of freshly

boiled, deionized water containing about 0.05 gram of Na2CO3. The solution has been kept in
tightly capped amber bottle. The formula weight of Na2S2O3.5H2O is 248.21 g mol-1.

Potassium iodate solution (approximately 0.065 mol L ): A 0.065 M potassium iodate

−1

solution has been prepared by dissolving about 1.39 g of pre-dried, primary standard of KIO3
solid to 500 mL mark of deionized water. This solution has been kept in 500 mL volumetric
flask. The formula weight of KIO3 is 214.00 g mol-1.

Note: Both of the reagents will be prepared by laboratory assistant.

B. Titration 1: Standardization of the sodium thiosulfate (Na 2S2O3) solution

1. Carefully, pipette 10.0 mL of the potassium iodate (KIO3) 0.065 M solution into a 250
mL Erlenmeyer flask.
2. Add approximately 20.0 mL of potassium iodide (KI) 0.2 M solution and 10.0 mL of
0.50 M sulfuric acid.
3. Swirl and immediately titrate with the thiosulfate titrant solution until the solution has
lost almost of its colour (straw yellow colour). At this point, add 2 mL of the starch
indicator and carefully complete the titration until the titrant removes the blue colour.
(Note: If the solution is clear and no blue colour is seen after addition of the starch, the
trial must be discarded and redone). This first titration is for rough titration.
4. Repeat the titration three more times for accurate titrations (titres agreeing within 0.10
mL) using the same procedure as above.

Note: All of these titrations should be performed carefully but rapidly to minimize air
oxidation of the iodide ion.

C. Titration 2: Analysis of ascorbic acid in the vitamin C tablets

1. Weight one tablet of 100 mg vitamin C and dissolve the tablet in 30 mL of 0.30 M
sulfuric acid (use 100 mL beaker). Use your glass rod to help break-up the tablet.
Some solid binding material of the tablet may not dissolve.
2. Filter the light orange solution into 250 mL Erlenmeyer flask. Discard the insoluble
material.
3. To the light orange solution (in the above 250 mL Erlenmeyer flask), add about 20.0
mL of KI 0.20 M solution and 10.0 mL of KIO3 0.065 M solution.
4. Swirl to mix and then carefully titrate with the standardized thiosulfate solution until
the dark brown solution changes to light brown solution.

5. Add this stage, add 2 mL of starch (the colour should change to dark brown-blue).

6. Continue the titration until the colour changes to the light orange colour. This first
titration is for rough titration.

7. Perform the titration three more times for accurate titrations (titres agreeing within
0.10 mL) with different tablet in each analysis. The formula weight of ascorbic acid is
176.14 g mol-1.

Results and Discussion

Standardization of the thiosulfate solution

i) Using manual calculation and Microsoft Excel, calculate the average volume of
sodium thiosulfate (Na2S2O3) solution from all three titrations, determine the standard
deviation (SD) and relative standard deviation (RSD).
ii) Form a table to report your data in two decimal places.
 iii) Calculate the molarity of the standardized sodium thiosulfate (Na2S2O3) solution
based on average value of titration volume.

Analysis of ascorbic acid in the vitamin C tablets

i) Prepare tables of titration data for the analysis of ascorbic acid in vitamin C tablets.
ii) Calculate the individual mass of ascorbic acid (vitamin C) found in each tablet in
milligrams.
iii) Calculate the average mass (in mg) of ascorbic acid found in all three tablets,
determine the standard deviation and relative standard deviation.

Part II: Determination of vitamin C content in juices and real-lemon

Materials and Method

Glassware and Apparatus

 Pipettes; 25 and 50 mL
 Burette; 50 mL and stand
 Conical (Erlenmeyer) flasks; 250 mL
 Measuring (graduated) cylinders; 10, 100, and 500 mL
 Beakers; 400 and 600 mL
 Volumetric flask; 250 mL

Chemicals
 Juice sample
 Real-lemon sample
 Potassium iodide (KI)
 Potassium iodate (KIO3)
 Vitamin C tablets
 Starch indicator (containing HgI2 as the preservative)
 Sulfuric acid; 3 M

Procedure

A. Reagents preparation

Iodine solution: Dissolve 5.00 g potassium iodide (KI) and 0.268 g potassium iodate (KIO 3)
in 200 mL of distilled water in a 400 mL beaker. Add 30 mL of 3 M sulfuric acid. Then pour
the solution into a 500 mL graduated cylinder, and dilute to a final volume of 500 mL with
distilled water. Mix thoroughly and transfer to a 600 mL beaker. Do not put this solution in a
volumetric flask!

Vitamin C standard solution (approximately 0.065 molL −1): Dissolve 0.250 g vitamin C in
100 mL water. Dilute to volume in a 250 mL volumetric flask.
B. Titration 1: Standardization of iodine solution

1. Add 25.0 mL of vitamin C standard solution into a 250 mL Erlenmeyer flask.

2. Add 10 drops of 1% starch solution.
3. Rinse your burette twice with 5-10 mL of iodine solution, and then fill it. Record your
initial burette volume.
4. Titrate the solution until the end-point is reached (the first sign of blue colour that
remains after at least 20 s of swirling). Record the final volume. This first titration is for
rough titration.
5. Repeat this titration at least three more times for accurate titrations (titres agreeing within
0.10 mL).
6. Use the Q-test check for bad data.

C. Titration 2: Analysis of vitamin C in juice sample

1. Add 25.0 mL of your juice sample solution into a 250 mL Erlenmeyer flask.
2. Add 10 drops of 1% starch solution.
3. Rinse your burette twice with 5-10 mL of iodine solution, and then fill it. Record your
initial burette volume.
4. Titrate the solution until the end-point is reached (the first sign of blue colour that
remains after at least 20 s of swirling). Record the final volume. This first titration is for
rough titration.
5. Repeat this titration at least three more times for accurate titrations (titres agreeing
within 0.10 mL).
6. Use the Q-test check for bad data.

D. Titration 3: Analysis of vitamin C in real-lemon sample

1. Add 10.0 mL of your real-lemon sample solution into a 250 mL Erlenmeyer flask.
2. Add 10 drops of 1 % starch solution.
3. Rinse your burette twice with 5-10 mL of iodine solution, and then fill it. Record your
initial burette volume.
4. Titrate the solution until the end-point is reached (the first sign of blue colour that
remains after at least 20 s of swirling). Record the final volume. This first titration is for
rough titration.
5. Repeat this titration at least three more times for accurate titrations (titres agreeing
within 0.10 mL).
6. Use the Q-test check for bad data.

Results and Discussion

Standardization of the Iodine solution

i) Prepare tables of the titration data for standardization of the iodine solution.
ii) Calculate the average volume of iodine solution from all three titrations, determine
the standard deviation and relative standard deviation.
 iii) Calculate the molarity of the standardized iodine solution based on average value of
titration volume.

Analysis of ascorbic acid in the juice and real-lemon

i) Discuss your Q-test result for the analysis of vitamin C in juice and real-lemon
samples.

ii) Prepare tables of titration data for the analysis of vitamin C of juice and real-lemon
samples.

iii) Based on the volume for each titration, calculate the amount of vitamin C in juice and
real-lemon samples including:

 moles of vitamin C present

 molarity of vitamin C
 concentration of vitamin C in g/L

iv) Using Microsoft Excel, calculate the average concentration of vitamin C, standard
deviation and relative standard deviation for both samples.

v) Draw bar chart consisting of average concentration for vitamin C in juice and real-
lemon samples. Include error bar for y-axis.

vi)Using manual calculation and Microsoft Excel, perform t-test at 95% confidence level
to compare the concentration of vitamin C contained in juice and real-lemon samples.
Discuss your finding.

Questions

1. What is the type of reaction that produces iodine from iodate and iodide?

2. Draw the structure of the organic compound of vitamin C.

3. Explain why the Na2CO3 need to be used in the preparation of thiosulfate solution.

4. Explain why the amber bottle that was used to store the thiosulfate solution needs to
be tightly capped.

5. A standard iodine solution was standardized against a 0.4123 g primary standard

As4O6 by dissolving the As4O6 in water solution. A small amount of NaOH solution
 was added to adjust the pH to 8. The titration of the sample requiring 40.28 mL iodine
solution. What is the concentration of the iodine solution?

As4O6(s) + 6 H2O 4 H3AsO3

H3AsO3 + I3- + H2O H3AsO4 + 3 I- + 2 H+

6. The purity of a hydrazine (N2H4) sample is determined by titration with iodine

solution. A sample of the oily liquid weighing 1.4286 g is dissolved in water and
diluted to I L in a volumetric flask. A 50.00 mL aliquot is taken with a pipette and
titration with the standard iodine solution in Question 5 requiring 42.41 mL titrant.
What is the percent purity by weight of the hydrazine?

N2H4 + 2 I3- N2 + 6 I- + 4 H+