Annals of the „Constantin Brancusi” University of Targu Jiu, Engineering Series, No.

3/2021

DETERMINATION OF VITAMIN C IN FOOD

BY THE TITRATION OF IODINE

Cazalbaşu Violeta Ramona

"Constantin Brâncuşi" University of Tg - Jiu, Faculty of Engineering

ABSTRACT: Vitamin C (ascorbic acid) is an essential antioxidant for human nutrition. Vitamin C deficiency
can lead to a disease called scurvy, which is characterized by abnormalities in the bones and teeth. Many fruits
and vegetables contain vitamin C, but cooking destroys the vitamin, so raw citrus fruits and their juices are the
main source of ascorbic acid for most people. Ascorbic acid is an organic acid with antioxidant properties,
involved in a number of processes that take place in living cells. In the human body vitamin C plays a complex
and important role; protects biologically active compounds from oxidative degradation, strengthens the immune
system, stimulates the biosynthesis processes of collagen, steroid hormones and neurotransmitters.

Keywords: vitamin C, iodine, food

1. INTRODUCTION No organ has ascorbate reserves as its

As a participant in hydroxylation,
vitamin C is required for the production of
collagen in connective tissue. These fibers are
ubiquitous throughout the body, providing it with
a stable but flexible structure. Some tissues have
a higher percentage of collagen in them,
especially the skin, mucous membranes, teeth
and bones. Vitamin C is needed for the synthesis
of dopamine, norepinephrine and adrenaline in
the nervous system or adrenal glands.
Vitamin C is also needed in the synthesis
of carnitine, which is important in the transfer of
energy to the mitochondria of cells. It is a
powerful antioxidant. The biological tissues with
the highest percentage of vitamin C content -
over 100 times the level of blood plasma - are the
adrenal glands, pituitary gland, thymus, corpus
luteum and retina. The brain, spleen, lung,
testicle, lymph node, liver, thyroid, small
intestinal mucosa, leukocyte, pancreas, kidney
and salivary glands have a concentration of
vitamin C 10 to 50 times higher than in plasma.
91
main function, so the body is very quickly left
without this substance if there is no continuous
intake to be absorbed by the digestive system.
Eventually, it can lead to death if the deficiency
is neglected.
The recommended nutritional intake of
vitamin C is 35 to 65 milligrams per day for
children, from 60 to 100 milligrams for adults
and adolescents. An increased intake of 120
milligrams per day is recommended for smokers.
This vitamin is easily oxidized and very sensitive
to heat and infrared and ultraviolet radiation.
Vitamin C deficiency, rare in
developing countries and exceptional in
industrialized countries. It is responsible for the
appearance of scurvy, a form of avitaminosis.
Caused by insufficient food intake, digestive
malabsorption, increased need or excessive
elimination, deficiency most often occurs in
elderly, alcoholic subjects, suffering from
chronic malabsorption or feeding without
vitamin C supplementation. It results in fatigue
and anemia, osteoarticular pain, edema,
gingivitis and tooth loss, bleeding, low
 Annals of the „Constantin Brancusi” University of Targu Jiu, Engineering Series, No. 3/2021
immunity. It was a condition very common
among sailors who went on long voyages or
during the winter, because fruits and vegetables
could not be kept fresh for a very long time.
The states of sub-deficiency (the stage
preceding the deficiency) would be much more
numerous, and currently the question is asked
about the possible relationship between a
deficiency in vitamin C intake and various
diseases (cancer, cardiovascular disease,
cataracts, etc.).
Vitamin C is prescribed orally in the
treatment of appropriate deficiencies, fatigue,
capillary and venous disorders, and intravenously
in case of methemoglobinemia (abnormal
increase in blood levels of methemoglobin, a
molecule unable to transport oxygen). In
contrast, contrary to popular belief, vitamin C
has no direct influence on the influenza virus, its
prophylactic action on influenza, the common
cold and any infections in general being
nonspecific, explained by stimulating anabolic
processes. Excessive intake of vitamin C (with
additional medication) can cause agitation and
insomnia, but it is not dangerous, the excess
being eliminated through urine and feces. For
doses equal to or less than 1000 milligrams per
day, there are no contraindications. Higher doses
should not be given in case of haemochromatosis
(disease due to iron accumulation in the body
tissues), oxalic kidney stones, gluco-6-phosphate
dehydrogenase deficiency or renal failure. It is
important to choose a suitable method for
preparing food to preserve its vitamin C content.
When cooking vegetables, the cooking time and
duration should be reduced and the water used in
cooking should not be discarded, for example by
steaming or soup cooking. Vitamin C in food is
identical to that in supplements. The structure of
vitamin C is very well understood, see ascorbic
acid, and there is no difference between the
benefits of natural and synthetic (although fruits
and vegetables also contain various other
nutrients, so vitamin C is not their only
advantage).
Recent observations suggest that the
effect of temperature and cooking on vitamin C
may have been overestimated: Because it is
water-soluble, vitamin C will dilute in cooking
92
water during the preparation of most vegetables -
but this does not mean that vitamin C is
destroyed. - She was still there, with the
difference that she went into the cooking water.
Not only the temperature, but also the
exposure time is significant. Contrary to what
was previously assumed (and still believed), it
takes much more than two to three minutes to
destroy vitamin C at boiling point. It also seems
that cooking does not extract vitamin C from all
vegetables as quickly; It has been suggested that
the vitamin is not destroyed when broccoli is
boiled. This can be due to a slower loss of
vitamin C in cooking water than other
vegetables.
Copper pans destroy vitamin C. Teas and
infusions with vitamin C supplements continue
to appear more and more on store shelves. Such
products would be pointless if the boiling
temperature did indeed destroy vitamin C at a
rate that had been previously suggested.
2. EXPERIMENTAL PART
One way to determine the amount of
vitamin C in food is to use a redox titration.
The redox reaction is better than an acid-base
titration because there are additional acids in a
juice, but few of them interfere with the
oxidation of ascorbic acid by iodine. Iodine is
relatively insoluble, but this can be improved
by complexing iodine with iodide to form
triiodide:
I 2 + I - ↔ I 3-
Triiodide oxidizes vitamin C to form
dehydroascorbic acid:
C 6 H 8 O 6 + I 3 - + H 2 O → C 6 H 6 O 6 + 3I -
+ 2H +
As long as vitamin C is present in the
solution, the triiodide is converted to iodide ion
very quickly. However, when all vitamin C is
oxidized, iodine and triiodide will be present,
which react with starch to form a blue-black
complex. The blue-black color is the end point of
the titration. This titration procedure is suitable
for testing the amount of vitamin C in vitamin C
 Annals of the „Constantin Brancusi” University of Targu Jiu, Engineering Series, No. 3/2021
tablets, juices and fresh, frozen or packaged fruits
and vegetables. Titration can be performed using
only iodine solution and not iodine, but the iodine
solution is more stable and provides a more
accurate result.
PREPARATION OF SOLUTIONS
Iodine solution
▪ Dissolve 5.00 g of potassium iodide
(KI) and 0.268 g of potassium iodate
(KIO3) in 200 ml of distilled water.
▪ Add 30 ml of 3 M sulfuric acid.
▪ Pour this solution into a 500 ml
graduated cylinder and dilute to a final
volume of 500 ml with distilled water.
▪ Mix the solution.
▪ Transfer the solution to a 600 ml
beaker. Label the glass as an iodine
solution.
Vitamin C standard solution
▪ Dissolve 0.250 g of vitamin C
(ascorbic acid) in 100 ml of distilled
water.
▪ Dilute up to 250 ml with distilled
water in a graduated flask. Label
the vial as a standard solution for
vitamin C.
Soluții de standardizare
▪ Add 25.00 ml of standard
vitamin C solution to a 125
ml Erlenmeyer flask.
▪ Add 10 drops of 1% starch
solution.
▪ Rinse the foam with a small
volume of iodine solution
and then fill it. Record the
initial volume.
▪ Titrate the solution until the
end point is reached. This
happens when you see the
first sign of blue that
persists after 20 seconds of
turning the solution.
▪ Record the final volume of
iodine solution. The
required volume was the
93
start volume minus the final
volume.
▪ Repeat the titration at least
twice. The results should be
0.1 ml.
HOW TO DETERMINE VITAMIN C
IN FRUITS AND VEGETABLES
• Weigh 15 g of the material to be
analyzed (eg onions, bell peppers, apples,
oranges, lemons, kiwis, etc.) with an accuracy
of 0.01 g.
• The weighed analyte is vigorously
ground with about 10 cm3 of 2% HCl solution
and 2.5 g of quartz sand or glass powder for
10 minutes.
• Quantitatively pass the mixture obtained
in a 50 ml volumetric flask and make up to
the mark with 2% HCl.
• Filter through a dry filter into a dry
vessel or centrifuge the suspension.
• Pipette 10 cm3 of filtrate into a 100 ml
Erlenmeyer flask, add 30 ml of distilled
water, 5 ml of KI, 5 ml of HCl and 1.5 ml of
starch solution.
• Titrate with a KIO3 solution to blue.
When potassium iodate is added to the
mixture, which also contains potassium
iodide, iodine is generated which oxidizes the
vitamin C present. After vitamin C has been
completely oxidized, the iodine generated will
form with the starch present an intense blue
colored inclusion complex. The color must
persist for 30 seconds.
• The analysis is repeated twice, the
average value of titrations is calculated (V in
ml)
CALCULATION OF RESULTS
Based on the equations of chemical
reactions, it is observed that one mole of
potassium iodate generates 3 moles of iodine,
which oxidizes 3 moles of vitamin C. The number
of moles of potassium iodate is: V * 0.0008 /
1000. The number of moles of oxidized ascorbic
acid in the sample analyzed by titration is then 3 *
V * 0.0008 / 1000
 Annals of the „Constantin Brancusi” University of Targu Jiu, Engineering Series, No. 3/2021
Since only 10 ml (of the 50 ml suspension
in the dimensioned flask) were introduced during
titration, it must be multiplied by 5. With the
molecular mass of vitamin C, of 176, the quantity
of vitamin C from the analyzed material is
obtained by multiplying initially weighed material
to be used in the extraction of vitamin C by
grinding): V * 0.0008 / 1000 * 5 * 176 The
vitamin C content is generally calculated in mg /
100 g of material, ie:
• Vitamin C mg% = V * 0.0008 / 1000 * 5
* 176 / G * 100 * 1000
• Vitamin C mg% = V * 0.0008 * 5 * 176
/ G * 100 = V * 70.4 / G
where:
V = no. ml KIO3 0.0008 M used for
titration;
G = Mass in g of material taken into
analysis.
BIBLIOGRAPHY:
1. 1. Marcel Avramiuc „Biochimie” vol.
I, University of Suceava Publishing
House,
94
2. 2. Neamţu G, Câmpeanu G, Socaciu
C, “Plant biochemistry (structural
part)”, Didactic and Pedagogical
Publishing House, R.A-Bucharest,
1993,
3. 3. I.F Dumitru „Biochemistry”,
Didactic and Pedagogical Publishing
House, Bucharest, 1980,
4. 4. S. Zinca „Animal Biochemistry”
Didactic and Pedagogical Publishing
House, Bucharest, 1971,
5. 5. I. Brunea and others „Plant
chemistry and biochemistry” Didactic
and Pedagogical Publishing House,
Bucharest, 1977,
6. 6. Ingerborg Theil-Tenchea “Plant
Biochemistry”, “Ştefan cel Mare
University” Publishing House,
Suceava, 1995,
7. 7. Dinu V and others „Medical
Biochemistry” Medical Publishing
House, Bucharest, 2002.