Abstract

The objective of the experiment is to determine the environmental factors that affected

the vitamin C content in orange juices.The experiment is carried out by using iodine solution.

The number of drops of orange juice used to react with all the iodine solution is recorded and

tabulated. From the result obtained, freshly squeezed orange juice contains the highest

amount of vitamin C compared to orange juices that are refrigerated, exposed to the sunlight,

and exposed to the air for an hour respectively. The average number of drops of freshly

squeezed orange juice used to react with all the iodine is the smallest which is 12 drops.

Environmental factors like air, sunlight, temperature have significant effects on the vitamin C

content in orange juice.

 Introduction

Vitamin C is a L-enantiomer of ascorbic acid, with chemical formula of C6H8O6, is a

water soluble vitamin which can be easily absorbed into our body. It is found in many

biological systems and foodstuffs, mainly from vegetables and fruits. It is essential in collagen

biosynthesis, osteogenesis, iron absorption and immune response activation. It is also reputed

as an excellent antioxidant due to its strong reducing properties.

It is a labile substance; therefore it is easily degraded by cooking and exposure to air.

Its oxidation is accelerated by the presence of heat and light. However, there are some

environmental factors negatively influence vitamin C concentration easily in fruit juices, such

as high temperature over certain values and freezing. Therefore this study is designed to

evaluate the effect of environmental factors on vitamin C concentration in fruit juice, mainly

about the duration of storage and effect of different temperature.

According to journal of ‘Ascorbic Acid Content of Commercial Fruit Juices and Its

Rate of Loss Upon Storage’, the highest values for ascorbic acid are obtained for natural juice

made by fruit squeezing. Upon exposure to air, ascorbic acid is oxidized by atmospheric

oxygen; the longer the exposure, the higher the oxidation time.

Besides, temperature will also affect the concentration of vitamin C. Analytical results

of the journal of temperature effects on vitamin C content in citrus fruit showed that the lower

the temperature the better the concentration of vitamin C in fruit juice. It is also supported by

the article of ‘ascorbic acid stability during refrigerated storage of orange–carrot juice treated

by high pulsed electric field and comparison with pasteurized juice’, the ascorbic acid

degradation rate in the juice stored at 2 °C was less than in the juice stored at 10 °C.
 Methodology

In order to achieve the objective which had outlined and mentioned above, we decided

to carry out the experiment by using iodine. Iodine has a brown colour when in solution and

iodide ions are colourless in solution. To make the iodine colour easier to see, starch is added-

this make a blue-black coloured complex. We can use this reaction to compare the amount of

vitamin C in different juices.

Before starting the experiment, the oranges are squeezed to obtain the juices. The

orange juices are equally divided into 20cm3 and pour into four beakers. All of the beakers are

labelled as A, B, C and D and will be put under different conditions. Beaker A is the freshly

squeezed orange juice, Beaker B will refrigerated for one hour whereas Beaker C and D will

exposed to sunlight and air for one hour respectively.

Firstly, 1 cm3 of starch solution is measured by using measuring cylinder and put into

a clean test tube. Then, 5 cm3 of distilled water is measured and added to the same test tube.

The test tube is shook gently. Next, 1 drop of iodine solution is added into the test tube by

using a dropper. A blue-black colour is appeared. A piece of white card is held as background

behind the test tube. After that, the freshly squeezed orange juice is added drop by drop by

using a clean dropper into the test tube. The test tube is shaken gently after every five drops

are added. The freshly squeezed orange juice is kept adding into the test tube drop by drop

until the blue-black colour has disappeared from view and no further change is observed. The

number of drops of freshly squeezed orange juice used is recorded. Excess drops of orange

juice are then added into the test tube to ensure that the blue-black colour has disappeared

completely.
 The experiment is repeated by using orange juices which are refrigerated, exposed to

the sunlight and exposed to the air for an hour respectively. All the results are recorded and

tabulated.

Result On the Vitamin C Content in Orange Juice

Results

Factors 1st 2nd 3rd Average

Freshly Squeezed Orange Juice 10 15 10 12

Orange Juice Is Refrigerated For 15 20 25 20

One Hour

Orange Juice Is Exposed To 25 35 30 30

Sunlight For One Hour

Orange Juice Is Exposed To Air 20 30 25 25

For One Hour

Figure 1: Number Of Drop Of Orange Juice Is Used To Decolourises Iodine Solution

 35

30

25

20

15

10

0
Freshly Squeezed Refrigerated Orange Orange Juices Exposed Orange Juices Exposed
Orange Juices Juices For One Hour To Sunlight For One To Air For One Hour
Hour

Graph 1: number of drops of orange juice is needed to react with iodine.

Discussions

Iodine is used to determine which environment factors affect the most vitamin c

content in orange juice in this experiment. I2 will form a blue complex in the presence of

starch (Vitex) and hence can act as its own indicator. Whilst the ascorbic acid is in excess, the

brown colour of I2 which is being reduced and decolourised. As soon as the ascorbic acid is

added into the test tube is being oxidized by the iodine present in the test tube, the iodine will

no longer be reduced and due to the Vitex that has been added, the solution becomes pale blue.

Therefore, an orange juice which has higher content of Vitamin C is needed in small amount

to react with the iodine. In addition, ascorbic acid in orange juice is readily oxidized and lost

depends on the condition of storage.

From the results obtained, we found that freshly squeezed orange juice contained the

highest amount of Vitamin C which only need 12 drops of orange juice to react completely

with iodine as compared to the orange juice that were refrigerated, exposed to sunlight, and
exposed to air for one hour. The content of Vitamin C in orange juice that was exposed in

sunlight for one hour is the lowest which was needed 30 drops of orange juice to reduce the

iodine in the test tube. This is probably due to the oxygen and light reacts with Vitamin C to

degrade it and this is accelerated when there are high temperatures as well.

Besides, ascorbic acid is a water soluble vitamin. When oxygen in the air reacts with

Vitamin C, oxidation occurs which is a chemical change that results in the breakdown of

Vitamin C. As temperature increases, the rate of chemical reactions will also increase and

some Vitamins C will be destroyed. Photodegradation will also occur when Vitamin C is

exposed to sunlight. When orange juice is exposed to air, ultraviolet ray from incidence light

causes the hydroxyl bond in ascorbic acid to break and being oxidised. Therefore, content of

of vitamin c is lowered.

According to the statement: juice with lots of ascorbic acid (Vitamin C) needs a small

number of drops to react with all the iodine and vice versa. In a nutshell, the highest content

of Vitamin C is the freshly squeezed orange juice, followed by the refrigerated orange juice,

orange juice is exposed to air and the lowest content must be the orange juice which is

exposed to sunlight.
 Conclusion

From the research that we had done, Vitamin C is easily oxidized and we had to

provide a good condition of storage to maintain the content of ascorbic acid in orange juice.

Besides, we found that the highest content of ascorbic acid is in the freshly squeezed orange

juice. Therefore, it is advised that we better consumed the orange juice after the juice is

squeezed.

Apart from that, the accuracy in finding the content of ascorbic acid in orange juice by

using iodine is lower as compare to the accuracy of using titration method.

 Reference

Aurelia Magdalena pisoschi et al; Ascorbic acid determination in commercial fruit juice

samples by cylic voltammetry, journal of automated methods and management in chemistry

volume 2008 (2008), article ID 937651

F.Torregrosa, M.J.Esteve, A.Frigola, C.Cortes, Journal of Food Engineering ,Volume 73,

Issue 4; April 2006, pages 339–345

Kabasakalis, V., Siopidou, D. and Moshatou, E. ; Ascorbic acid content of commercial fruit

juices and its rate of loss upon storage 2000; Food Chemistry 70 (3): 325-328.

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