Chemistry Investigatory Project-Class XII

Topic:- WHY DO APPLES TURN BROWN ON EXPOSURE TO AIR?

Introduction: Apples have been found to have very strong antioxidant activity,
inhibit cancer cell proliferation, decrease lipid oxidation, and lower cholesterol.
Apples contain a variety of phytochemicals, including quercetin, catechin,
phloridzin, and chlorogenic acid, all of which are strong antioxidants. The
phytochemical composition of apples varies greatly between different varieties of
apples, and there are also small changes in phytochemicals during the maturation and
ripening of the fruit.
Fruit and vegetables have health benefits for consumers, due to their content of
fibre, vitamins, and antioxidant compounds. However, for the antioxidant
compounds, many changes occur during harvesting, preparation (fresh-cut fruits),
and storage of these fruits. These changes induce a pronounced loss of
microbiological and antioxidant qualities. Thus, preservation against oxidation
in food during processing and storage has become an increasing priority in the food
industry. Oxidation is the second most important cause of food deterioration
after that induced by microbiological contamination. The main oxidative reactions
are enzymatic browning. They involve two enzymes: polyphenol oxidase (PPO)
and peroxidase (POD). PPO catalyzes two reactions; the first is the hydroxylation of
monophenols to diphenols, which is relatively slow and results in colourless products.
The second, the oxidation of diphenols to quinines, is rapid and gives coloured
products. The substrates involved in these reactions are located in the vacuoles while
enzymes are in the cytoplasm; the reactions can take place only if they are mixed and
in the presence of oxygen. So, all phenomena (cutting, shock, loss of firmness) lead to
the starting of browning reactions which induce losses or changes of flavour, odour,
and nutritional value. To avoid this phenomenon various methods are developed.
The role of these methods is either to inactivate polyphenol oxidase (PPO) or to
avoid contact between the enzyme and its substrate, either by adding antioxidants or
by maintaining the structural integrity of the food.
AIM: To study the chemical reaction behind the browning of apples.

Apparatus/Materials Required:
Apple slices
Diluted lemon juice
Salt water
Baking soda water
Vinegar
Milk
Small containers
Theory: This unappetizing phenomenon is actually due to a chain of biochemical
reactions known as “enzymatic browning.” When an apple is injured (or cut into
pieces), the plant tissue is exposed to oxygen. This triggers an enzyme known as
polyphenol oxidase (PPO) to oxidise polyphenols in the apple’s flesh. This results
in new chemicals (o-quinones) that react with amino acids to produce
brown-coloured melanins. This reaction of enzymatic browning occurs readily at
warm temperatures when pH is between 5.0 and 7.0. The presence of Iron(Fe) or
Copper(Cu) can increase the rate of reaction. This can be observed when the fruit is
cut with a rusty knife or mixed in a copper bowl. Different apple varieties contain
different amounts of both the initial enzyme and the polyphenols; thus, they brown
at different rates. Bruising or other injury to the plant tissue disrupts the arrangement
of chemicals within the cells and allows these chemicals to make contact with oxygen.
This may lead to browning of uncooked fruits. Enzymatic browning can be a
significant problem, limiting the shelf life of many fruits and vegetables which have
had little heat applied during processing. However, enzymatic browning is not
always a defect. The browning reaction contributes to the desirable colour and
flavour of raisins, prunes, coffee, tea and cocoa.
Enzymatic browning is not unique to apples. Pears, bananas, and eggplants also turn
brown reasonably quickly when cut. Enzymatic browning is also responsible for the
desirable dark colour of prunes, coffee, black tea, and cocoa. To reduce browning in
apples, keep the slices refrigerated to slow the reaction. You can also coat them with
lemon or pineapple juice. The acids in these juices slow the reaction, and the
antioxidants inhibit it. If you don't mind the added sugar, covering the slices with
honey, caramel, or sugar syrup effectively blocks their exposure to oxygen.

Treatment of apple slices with ascorbic acid, citric acid, or acetic acid will reduce the
level of browning. The reduction in browning is dependent on the type of substance
and its concentration. Soaking in water will temporarily reduce the level of browning
by restricting the amount of oxygen in contact with the apple slices.

Procedure:
1. Cut out small pieces of paper and write the name of the liquids you are going to
test. One of them should be “air” for your control variable.
(Having a control variable is necessary so that you can compare it to the other apples
that have been submerged in various liquids.)

2. Pour the other liquids into separate containers.

3. Place the labels next to the corresponding containers.

4. Place an apple slice in each container so that it’s submerged in the liquid.

5. Let the apple slices sit for a few minutes in the respective containers.
6. Place the apple slices on a plate and put the corresponding labels next to them.

7. Wait for about an hour and check on the apples. Discuss the progress of the
oxidation.

8. Wait for a couple more hours and check on the apples. Discuss any new
observations.

9. Leave your apple slices for a total of 5 hours.

Observations:

Lemon juice- The apple slice dipped in lemon juice did not show any browning.
This is because of the low pH value of lemon juice(between 2-2.5) which prevented
the apple from turning brown. The acid in lemon juice inactivates polyphenol
oxidase to prevent browning.

Salt water- Salt water prevented the oxidation of the apple slice because it
surrounded the apple cells with a saltier environment than the inside of the cells. As a
result, water moves outside of the cells through osmosis in an attempt to equalise the
concentration of salt and water inside and outside of the cell. The water and salt on
the surface of the apple slice interfere with oxygen reaching the fruit’s surface and
therefore slowing down oxidation.

Baking Soda Water- Baking soda, which has a high pH value, made the apple
brown faster. This is because substances with a high pH contain higher amounts of
oxygen. Therefore, covering the apple with baking soda solution actually brought
more oxygen to the surface of the apple, accelerating the browning.
Vinegar- Apple Slices dipped in vinegar turned out to be slightly brown. Even
though vinegar has a pH between 2.4-3.0, it seemed to have accelerated the browning
slightly despite being acidic.

Milk- Milk is not acidic and apple slices turn brown after some time. It was able to
stop browning for a little time by coating the apple slice and coming in contact with
air. The apple slices turned less brown in the experiment with milk.

The control was not treated with any solution and therefore the reaction was able to
proceed and the fruit turned brown.

Conclusion:
According to our results, lemon juice had the lowest browning effects across all time
periods, and the vinegar treatment group had the highest browning effects across all
time periods. However, other treatment groups showed various browning effects
across time. At time zero, all treatment groups showed better delaying browning
effects in comparison to the control group. Thus, we can suggest that all five
treatment groups will be effective for delaying browning effects within a short period
of time, such as an hour. From 4 hours to 6 hours, only the vinegar group had much
more severe browning effects than the control group. This suggested that lemon
juice, saltwater, and milk, could all be potential ways to delay browning effects in the
relatively long term, but some of the effects might be too small to be statistically
significant.
Pretreatment methods:
1. An effective method is Sulfuring. Sulfuring commercially is done on fruits using
sulphur dioxide gas
2. A sulfite dip can also be used. Either sodium bisulfite, sodium sulfite, or sodium
meta-bisulfite can be used; use 3/4 to 1.5 teaspoons of sodium bisulfite per quart of
water. Place sliced fruit in the mixture and soak for 5 minutes. If the fruit is in halves,
place in the mixture for 15 minutes. Rinse lightly under cold water and place on
drying trays. This can be done indoors or outdoors.
3. A fruit juice dip can also be used. Any fruit juice that is high in vitamin C is an
effective pretreatment though it does not work as well as ascorbic acid (remember,
ascorbic acid is pure vitamin C). Examples are orange, lemon, pineapple, and grape
juice.
4. Honey dip — Many store bought dried fruits have been dipped in a honey
solution. Honey dipped fruit is much higher in calories
In recent years, valuable progress has been made in understanding
polyphenol oxidases and polyphenol of apple and apple products. However many
studies have been restricted to model systems with and enzyme acting on a single
phenolic substrate. There is a great need for studies of enzymatic action on mixtures
of phenols and of non-enzymatic reactions that follow o-quinone formation, as these
studies would provide a better understanding of the relationship between the extent
of browning and the phenolic composition of fruit and vegetables.