Executive Summary of Minor Research Project on

Analysis of Vitamin C in Commercial and

Natural Fruit Juices

Fruits are not only delicious but healthful too. Botanically, fruits
are the part of the plant that carries the seeds for future generations of
plants. They are often attractively 'packaged' by nature to encourage
animals and birds to eat them and scatter the seeds. Rich in vitamins A
and C, plus folate and other essential nutrients, they may help prevent
heart disease and stroke, control blood pressure and cholesterol, prevent
some types of cancer and guard against vision loss. Fresh fruits were
Nature's greatest gifts. They were a unique group of foods because they
have a wide variety of types, flavors, colors, and textures. Relatively
recent preservation techniques and modern transport facilities have
increased the variety of fruit available.

. Vitamin C is one of the most important vitamins for human

nutrition that is supplied by fruits and vegetables. Citrus fruits
(especially oranges, lemons, and grape fruits), gooseberries, strawberries,
and raspberries provide a good source. Vitamin C or ascorbic acid is an
essential, water soluble organic compound important in animal
metabolism. L-Ascorbic acid (AA) is the main biologically active form
of vitamin C. Ascorbic acid is reversibly oxidized to form L-
Dehydroascorbic acid (DHA), which also exhibits biological activity.
Since DHA can be easily converted into AA in the human body it is
important to measure both AA and DHA in fruits and vegetables to
know vitamin C activity (Lee and Kader, 2000).Among the vitamins,
vitamin C (ascorbic acid) is an essential micronutrient required for
normal metabolic function of the body ( Jaffe, 1984). Vitamin C is
easily oxidized, and the majority of its functions invivo rely on this
property. Most animals produce it in their bodies, but humans, other

1
primates, guinea pigs lack an enzyme called Gulonolactone oxidase, that
convert glucose to Vitamin C. So they lack the ability to synthesis
Vitamin C in their body and need to take it through their diet. A vitamin
C deficiency in humans results in the disease called scurvey, whose
symptoms include hemorrhaging, joint pain and exhaustion (Brody,
1994 and Pauling, 1976).A very small daily intake of vitamin C (10-15
mg/day for an adult) is required to avoid deficiency and stave off
scurvey (Kallner,1986).

Vitamin C is the major water-soluble antioxidant within the body

(Sies, et al., 1995; Levine, 1986; Levine, 1995). It lowers blood pressure
and cholesterol levels ( Rath, 1993). Numerious analysis have shown
that an adequate intake of vitamin C is effective in lowering the risk of
developing cancers of the breast, cervix, colon, rectum, lung, mouth,
prostate and stomach (Levine, 1996; Block, 1992; Block, 1991).
Vitamin C is generally non-toxic. For maintaining a good and sound
health and for prevention from common cold, human body should be
kept saturated with vitamin C. Vitamin C toxicity is very rare, because
the body cannot store the vitamin however ,amounts greater than 2000
mg/day are not recommended because such high doses can lead to
stomach upset and diarrhea. Keeping in view its importance, the
estimation of Vitamin C containing fruits assumes significance.

Citrus species (Rutaceae), the most popular fruits, originated in

South-East Asia and then gradually spread to different parts of the
world. These fruits contain a variety of sugars, citric acid, ascorbic acid,
carotenoids, minerals, essential oils, etc and play an important role in
human nutrition as excellent source of antioxidants (ascorbic acid,
carotenoids and phenolic compounds). These constituents are considered
to be essential components of functional foods. Many of these
substances prevent damage to cell membrane and other structures by
neutralizing free radicals. Ascorbic acid is the most important
antioxidant in citrus fruit juices and it protects the organism from
oxidative stress. Vitamin C cannot synthesize through body cells, nor
does it store it. It is therefore important to include plenty of vitamin C
containing foods in daily diet.

2
 One of the objective of the present study was to relate the content
of ascorbic acid of six Citrus fruits namely Citrus sinensis (Orange),
Citrus limon (Lemon),Citrus paradisi (Grape), Citrus maxima (Babloos
naranga), Citrus limetta (Mosambi) and Punica granatum (Pomegranate)
with the view of making recommendations for their intake. A redox
titration, involving an Iodometric method, has been used to do the
analysis.
Various reports have shown fruits to be excellent sources of
vitamin C. But it is lost from foods during preparation, cooking or
storage. To find out the effect of temperature on stability of Vitamin C a
temperature dependence study of vitamin C was carried out under three
temperature regimes representing the ranges the fruits may be exposed
to during processing and storage. The PH values of selected fruit juice
samples were also determined.
The pH of citrus fruit juices selected ranged from 2.60 to 4.10
and was found to be lesser in case of lemon fruits when compared to
others.PH values of fruit juices are different due to the variation in
amount of acidic contents. All samples were found to be acidic whose
intake generates acidic environment in the stomach.
The result of the average value of vitamin C in each fruit juice
samples under the specified condition are tabulated in Table below. It
showed the highest concentration of vitamin C found in Babloos juice,
hitting (31.97) mg /100g, the lowest level was found in Grape juice,
when it reached (2.34)mg /100 g of juice. The amount of Vitamin C in
juices of six different citrus fruits (Orange, Mosambi, Pomegranate,
Babloos, Grape and Lemon) is as follows.

Bambloos > Orange >Mosambi>Lemon>Pomegranate>Grape

3
No Fruit Juice Condition Temperature Total Vitamin C
Samples (mg/100g)

1 Orange Fresh 320C 31.24

2 Lemon Fresh 320C 16.56

3 Grape Fresh 320C 2.3409

4 Babloos Fresh 320C 31.97

5 Mosambi Fresh 320C 30.506

6 Pomegranate Fresh 320C 12.77

Table 2: Total Vitamin C Content in Natural Fruit Juice Samples

The effect of temperature on the amount of Vitamin C in citrus fruits

was also calculated by titrating the juices using iodine solution. It can be seen
from analytical results in table 3 that the lower the temperature the better the
concentration of Vitamin C in fruit juice.

Concentration
No Fruit sample At room At 600C At 900C
temperatu
re (mg/g) (mg/g) (mg/g)

1 Orange 31.24 29.507 22.59

2 Lemon 16.56 14.0348 3.1958

3 Grape 2.3409 2.1608 1.639

4 Babloos 31.97 28.071 23.096

4
 5 Mosambi 30.506 28.229 24.35

6 Pomegranate 12.77 12.77 9.295

Table 3: Total Vitamin C Content in Natural Fruit Juice Samples at

different temperature

Higher temperature does not favour Vitamin C. it is better to

maintain or store Vitamin C in a place below the room temperature. This
is consistent with reports that, climate, especially temperature affect
vitamin C level. Areas with cool nights produce citrus fruits with higher
vitamin C levels. Hot tropical areas produce fruit with lower levels of
vitamin C. Vitamin C loss during storage depends on the type of storage
method employed, for example, handling and storage; oxygen is the
most destructive ingredient in juice, causing degradation of vitamin C.
Juice should be discouraged from being display in the hot weather above
room temperature in order to maintain production concentration.
Our temperature dependence study on citrus fruits were found to
follow a similar pattern of loss. This is because the vitamin C is more
sensitive to temperature. Degradation was observed high in our selected
samples as the temperature was raised to 90oC

Conclusion
Determination of ascorbic acid content by iodometric titration is
an easy, safe, and fast method. The redox reaction is preferable to an
acid-base titration because a number of other species in juice can act as
acids. This would help in quickly determining an estimate of ascorbic
acid content of fruits for the purpose of recommending them for
consumption to curb deficiency problems. Adequate consumption of
fruits with high level of vitamin C can help in health improvement and
thus reduce diseases such as diabetes, glaucoma, atherosclerosis, stroke,
heart diseases and cancer The ascorbic acid content of the juices of six

5
Citrus fruits – Orange, Mosambi, Pomegranate, Babloos, Grape and
Lemon were determined iodometrically in order to know which fruit
would best supply the ascorbic acid need for the body. Results showed
that Bambloos had the highest value of ascorbic acid, 31.97mg/100g
followed by Orange, 31.24mg/100g and then Mosambi, 30.50mg/100g.
Grape had the least value, 2.34mg/100g. It therefore follows that
Bambloos would supply more ascorbic acid per 100gram for body need
compared to the other fruits.
Bambloos > Orange >Mosambi>Lemon>Pomegranate>Grape
Temparature effects on Vitamin C content in citrus fruits were
also determined using iodometric titration method under three
temperature regimes (room temperature, 60oC and 80oC), representing
the range of temperatures the fruits may be exposed to during processing
and storage. It was observed that Vitamin C content was decreased as it
was exposed to higher temperature. The decrease was observed high
when the temperature of the juice was raised and kept at 60oC. This is as
a result of increase in oxidation of ascorbic acid with increase in
temperature, as higher temperature favours redox reaction. This work helped
to demonstrate the effects of processing and storage on the ascorbic acid
contents of these fruits. Also lower pH value was preferred to prolong
the shelf life.
Vitamin C exist in the form of drugs as swallow tablets, chewing
tablets, swallow capsules, solvents and injection. We can had access to
the handling of the recommended amount of vitamin C easily through
alternative medicine but the damage will be limited in the short term if
they are to stop dealing dose, therefore preferred experts dealt vitamins
by natural eating and drinking, and not to rely on synthetic substitutes.
Our project here makes some suggestions for the preferential intake of
fruit juices.

6. References
1. Jain, J. L., Jain, Sunjay and Jain, Nithin, Fundamentals of Biochemistry, S.
Chand and Company Ltd., Ram Nagar, New Delhi, pp. 1009–1010.
2. Swaminathan, M., Handbook of Food and Nutrition, The Bangalore
Printing and Publishing Co., Ltd, pp. 58–60.

6
3. Nay, S., Project Done on Vitamin C Contents of Citrus Fruits and their
Products.
4. Ashwah, F.A. et. al, Project Done on Effect of Stage of Maturity on the
Properties of Orange JuiceConcentrates.
5. Andrew, et. al, Project Done on, “Stability of Ascorbic Acid in Orange
Juice Exposed to Light & Air.”
6.Arya, S.P., Mahajan, M., and Jain, P. Photometric methods for the
determination of Vitamin C. Analytical Sciences. 1998: 14; 889-895.
7. Brody, T. Nutritional Biochemistry; Academic Press: san Diogo, CA. 1994;
pp. x and 450-9.
8. Block, G. Epidemiologic evidence regardingvitamin C and cancer.
American Journal of Clinical Nutrition. 1991: 54; 1310S-14S.
9. Block, G. The data support a role for antioxidants in reducing cancer risk.
Nutrition Reviews. 1992: 50(7); 207-13.
10.. Biswas S.K. and Mannan M.A. Determination of vitamin C (ascorbic acid)
in some fruits and vegetables; B. J. Sci. & Ind. Res. 1996: 1; 31.
11. Carpenter, K.J., 1986. The history of scurvy and vitamin C. Cambridge:
Cambridge University Press.
12. Roberts, J.D. and M.C. Caserio, 1977. Basic Principles of Organic
Chemistry. California: W. A. Benjamin, Inc.
13. Ohio State University, Department of Human Nutrition and OSU
Extension 2004. Vitamin C (Ascorbic Acid)., HYG-5552-05.
14. Ajibola, V. O., Babatunde, O. A. and Suleiman, S. 2009. The Effect of
Storage Method on Vitamin C Content in Some Tropical Fruit Juices. Trends
in Applied Sciences Research. 4(2), pp. 79-84.
15. Arya, S. P. and Mahajan, M. 1997. Colorimetric Determination of
Ascorbic Acid in Pharmaceutical Preparations and Biological Samples.
Microchim. Acta. 127, pp. 45-49.
16. Arya, S. P., Mahajan, M. and Jain, P. 2000. Non-spectrophotometric
methods for the determination of Vitamin C. Analytica Chimica Acta. 417, pp.
1-14.

**************************

7
8