DETERMINATION OF ASCORBIC ACID (VITAMIN-C)

AIM:
To determine the weight of Ascorbic acid (vitamin-C) present in the given formulation
sample namely CELIN - 500mg tablets.

PRINCIPLE:
Ascorbic acid, commonly known as vitamin-C is one of the essential vitamins for human
beings. Chemically it is related to monosaccharides. Its molecular formula is C6H8O6. It
is a reducing agent and is readily oxidized by oxidizing agents such as Potassium bromate
(KBrO3), Iodine etc, to Dehydroascorbic acid. When Ascorbic acid reacts with Iodine, the
following reaction takes place.

C6H8O6 + I2 → C6H6O6 + 2H+ + 2I−

ASCORBIC ACID DEHYDRO ASCORBIC ACID

OH
HO

O O
O O
HO HO

HO OH O O

ASCORBIC ACID DEHYDRO ASCORBIC ACID

Ascorbic acid can be determined by direct titration with standard Iodine solution or by
back-titration method, in which an aliquot sample of Ascorbic acid is treated with a known
excess of Iodine solution; when all the Ascorbic acid content of the sample solution is
quantitatively converted into Dehydroascorbic acid. The excess of unreacted Iodine
solution is back-titrated by standard Sodium thiosulphate solution, using starch indicator.

Ascorbic + I2 solution → Dehydro ascorbic acid + I2 solution

(A known excess) (un reacted)

I2 + 2S2O32-  2I − + S4O62-
(un reacted) tetra thionate
At the equivalence point
 meq I2 ≡ meq of Ascorbic acid + meq of Sodium thiosulphate

From the above titration reaction, it is clear that the equivalent weight of Ascorbic acid is
½ of the formula weight. (The formula weight of Ascorbic acid is 176.14 g/mol).

Since Sodium thiosulphate is not a primary standard substance, the solutions of Sodium
thiosulphate are to be standardized by using standard Potassium dichromate or standard
copper (II) solution such as standard copper sulphate solution.

Potassium dichromate reacts quantitatively with Potassium Iodide in the presence of

mineral acid medium and liberates an equivalent amount of Iodine. The liberated Iodine
can be titrated with Sodium thiosulphate solution, when the following reaction takes
place.
Cr2O72- +14H+ +6e-  2Cr3+ +7H2O
6I-  3I2 + 6e-
________________________________________________________
Cr2O72- + 6I- +14H+  2Cr3+ + 3I2 +7H2O
________________________________________________________
I2 + 2S2O32-  2I- + S4O62-
tetrathionate ion

From the above titration reaction, it is clear that, at the equivalence point,

1 mmol Cr2O72- ≡ 3 mmol I2 ≡ 6mmol Na2S2O3

Therefore
1 mmol Cr2O72- ≡ 6 mmol Na2S2O3

The equivalent weight of Potassium dichromate is 1/6 of its formula weight and that of
Sodium thiosulphate is same as its formula weight.

The end point in Iodometric titrations is determined based on the principle that starch
solution reacts with Iodine to form an intense blue coloured complex and is visible at a
very low concentration of Iodine. At the end point, Sodium thiosulphate reacts with
Starch-Iodine complex and liberates free Starch into the solution. Since starch is
colourless, the colour change at the end point is from dark blue to colourless in case of
simple Iodine solutions. However, due to the presence of hydrated Cr (III) ions, the colour
of solution becomes the colour of hydrated Cr (III) ions, i.e. emerald green cololur.
Therefore, the colour change at the end point is from grayish blue to the emerald green.
REAGENTS REQUIRED:
K2Cr2O7 AR Crystals, ~ 0.1 M Sodium thiosulphate solution, Solid Sodium
hydrogencarbonate, Conc.HCl, 10% KI solution,Glacial Acetic acid and Starch indicator

PROCEDURE:
PREPARATION OF STANDARD POTASSIUM DICHROMATE SOLUTION:
Weigh out accurately about 1.25 g of Potassium dichromate AR crystals into a 250 mL
volumetric flask. Dissolve the contents in de-ionized water and make up the solution to
the mark with de-ionized water. Shake the contents well and calculate the concentration
of Potassium dichromate solution.

STANDARDIZATION OF SODIUM THIOSULPHATE SOLUTION:

Rinse and fill the burette with the Sodium thiosulphate solution to be standardized.
Pipette out 20 mL of the standard potassium dichromate solution into a 250 mL conical
flask. To this add about 1 g of solid sodium bicarbonate and 6 mL of Conc HCl solution.
Swirl the contents to dissolve the solid, when the liberated CO 2 will displace air in the
flask. There by it prevents aerial oxidation of KI. Now add 15 mL of 10% KI solution, and
swirl the flask and stopper it. Allow the flask to stand for 2 minutes in the dark. Rinse the
stopper into the flask. Dilute the contents by adding about 20 mL of de-ionized water.
Titrate the liberated Iodine with Sodium thiosulphate solution until a pale greenish yellow
colour is obtained. At this stage, add 2 mL of Starch indicator and rinse down the inner
sides of the flask with de-ionized water. The colour of the solution becomes dark blue due
to the formation of Starch-Iodine complex. Continue the titration with constant swirling
until one drop of the titrant changes the colour from dark blue to emerald green. The
green colour of the solution is due to the hydrated Cr (III) ions. Record the burette reading
and repeat the titrations with fresh aliquots of the standard Potassium dichromate
solution. Tabulate the results and calculate the concentration of the Sodium thiosulphate
solution.

STANDARDIZATION OF IODINE SOLUTION:

Pipette out 20 mL of I2 solution into a 250 mL conical flask. Dilute the contents to about
100 mL by adding de-ionized water. Titrate the contents against std Sodium thiosulphate
solution until it turns to pale yellow. At this stage, add 2 mL of starch indicator and
continue the titration until the solution just becomes colourless. Record the burette
reading and repeat the titrations with fresh aliquots of I2 solution. Tabulate the results
and calculate the concentration of Iodine solution.
PREPARATION OF SAMPLE SOLUTION:
Place the given Ascorbic acid formulation sample i.e. CELIN-500 mg tablet into a 250
mL volumetric flask. Dissolve the tablet in about 50 mL of de-ionized water and swirl the
flask to hasten the dissolution. A small amount of binder and other inert material present
in sample may not be dissolved and remain as suspension. Fill the flask with de-ionized
water to the mark and shake the contents well.

TITRATION OF UNREACTED IODINE:

Transfer 20 mL of the sample solution into a 250 mL conical flask and add 20 mL of
standard Iodine solution (0.05 N I2 solution); when the Ascorbic acid content of the
sample solution reacts quantitatively with an equivalent amount of Iodine and the excess
unreacted Iodine remains in the solution. Titrate the unreacted Iodine against std. Sodium
thiosulphate solution until it turns pale yellow. At this stage add 2 mL starch indicator
and continue the titration till the solution becomes colourless. Record the burette reading
and repeat the titrations with fresh aliquots of sample solution. Tabulate the results and
calucate the weight of Ascorbic acid present in the whole of the given Ascorbic acid
formulation namely CELIN-500 mg tablet.

RESULT:
The given Ascorbic acid formulation i.e. CELIN-500 mg tablet contains _______ mg
Ascorbic acid.