Aim

To study and calculate the content of ascorbic acid in different citrus fruits.

Theory
Vitamin C or ascorbic acid in citrus fruits is a water soluble carbohydrate like substance
involved in certain metabolic processes of animals. Although most of the animals can
synthesise vitamin C, it is necessary in the diet of some including men and other primates. In
order to prevent scurvy, disease which is haemorrhage especially in skin and mucous
membranes Vitamin C was identified as a curative agent for survey in 1928. The name
ascorbic acid is derived from the expression anaisearbatic vitamin, referring to vitamins
ability to prevent and to curve scurvy. First discovered in 1928 by Hungarian biochemist and
Nobel Prize winner Albert Seent Ceyorghi. Vitamin C has been the object of continued active
laboratory research to determine the specific mechanism of action of cells.
Structure

Ascorbic acid (AA) is a simple sugar. It is the most active reducing agent known to occur
naturally in living tissues, and is easily reversibly oxidized to Dehydroascorbic acid (DHA).
Ascorbic acid is a white crystalline stable substance, when in dry light and air. It is easily
soluble in water and easily oxidized especially in alkaline medium and exposure to heat and
light, it reacts with metals, particularly copper. It is fairly soluble in cold acid solution.
Though the first stage in its oxidation to DHA is reversible, oxidation to diketo-gulnic acid
cannot be reversed.

Chemistry
1. ACIDITY
Ascorbic acid, the formula of which C6H8O6, behaves as a vinologous carboxylic acid, There
are two resonance structures for the deprotonated form, differing in the position of double
bond.
Another way to look at ascorbic acid is to consider it an enol. The deprotonated form is an
enolate, which is usually strong basic. However, adjacent double bond stabilized the
deprotonated form.
2. TAUTOMERISM
Ascorbic acid is rapidly in converts into two unstable diketone tautomers by proton transfer,
although it is the most stable in the enol form. The proton of the enol is lost, reacquired by
electrons in the form of double bond, to produce a diketone. It is an enol reaction there are
two possible forms: 1,2-diketone and 1,3-diketone.
3. DETERMINATION
The concentration of the solution of ascorbic acid can be determined in many ways; the most
common way involves titration with an oxidizing agent.
DCPIP
A commonly- used oxidizing agent is the dye 2,6-dichlorophenol indophenols, or DCPIP
for short. The blue dye is run into the ascorbic acid solution until a faint pink colour persists
for 15 seconds.
IODINE
Another method involving using iodine and a starch indicator, wherein iodine reacts with
ascorbic acid, and when all the ascorbic acid had reacted, the iodine is excess, then forming a
blue black complex with starch indicator. This indicates the end point of the titration. As an
alternative, ascorbic acid can be reacted with iodine in excess, followed by back titration with
sodium thiosulfate while using starch as indicator
IODATE AND IODINE
The above method involving iodine requires making up and standardizing the iodine solution.
One way around this is to generate the iodine in presence of ascorbic acid by the reaction of
iodate and iodide ion in acid solution.

N BROMOSUCCINIMIDE
A much-less-common oxidizing agent is N bromosuccinimide, (NBS). In titration the NBS
oxidizes the ascorbic acid 9 in presence of potassium iodide and starch). When the NBS in
excess (i.e., the reaction is complete). The NBS liberates iodine from the potassium iodide,
which then forms the blue/black complex with starch, indicating the endpoint of the
titration.

Uses
Ascorbic acid is easily oxidized and so is used a reductant in photographic developer
solutions amongst ethers and as a preservative. Exposure to oxygen, metals, light, and heat
destroys ascorbic acid, so it must be stored in dark, cold and not metal container. The Lenantiomer of ascorbic acid is known as vitamin C. The name ascorbic comes from its
property of preventing and curing scurvy. Primates including humans and few other species
of the same animal kingdom, notably the guinea pig, have lost the ability to synthesize
ascorbic acid, and must obtain it in their food. Ascorbic and its sodium, potassium, calcium
salts are commonly used as antioxidant food additives. These compounds are water-soluble
and thus cannot protect fact from oxidation. For this purpose, the fat soluble esters of
ascorbic acid with long chain fatty acid can be used as food antioxidants. 80% of Worlds
supply of ascorbic acid is produced in China
Requirements
Water
Spirit
Starch
Iodine(solid)
Lemon Juice
Orange Juice
Citrus Maxima Juice
Burette
Pipette
Conical flask
Standard flask
Bunsens burner
Muslin cloth
Procedure
Weight 0.254g of solid iodine and pour in a dry beaker. Add 4g of solid potassium iodide.
Then add distilled water then dissolve iodine and potassium in it. Transfer this solution to a
clean 100ml volumetric flask and prepare required quantity of distilled water that was added
to make 100ml of iodine solution. In this way another 100ml of iodine solution is prepared.
This solution has a molarity of 0.01M.

Now a starch solution is prepared by adding a spatula of starch to 100ml of water and
subsequently boiling it.
The fruit juices are extracted and filtered using muslin cloth.
The iodine solution of 0.01M is taken in a burette and 5ml of filtered juice is pippeted out in
a conical flask. To the juice 1ml of starch solution is added.
The solution of juice is titrated against iodine solution. The process is stopped at the point
the colour of solution in conical flask changes from fruit juice to violet colour.
Three concordant readings are taken.
Result
The following are the percentage of ascorbic acid to various citrus fruits.
Lemon juice = 31.68%
Orange juice = 49.28%
Citrus Maxima = 70.4%
Conclusions:
The percentage of ascorbic acid was found to be less in lemon juice and more in Citrus
Maxima juice. Here starch solution was added to the fruit juices, before doing titration due to
which some of the fruit content reacted starch and a permanent complex was formed which
could not be oxidized

Bibliography
Sarawathi lab manual chemistry XII
Comprehensive lab manual chemistry
www.britanica.com
www.google.com
www.wolframalpha.com