EXPERIMENT 6

ENZYMATIC BROWNING
OBJECTIVE:
1. To determine the enzymatic browning in apples by using several
treatments
2. To determine the time taken for fruit to turn brown.
3. To determine the control of browning reaction.
4. To determine the acceptability of enzymatic browning in apples
including colour, odour and texture.
INTRODUCTION
Enzymatic browning is one of the most important reactions that occur
in fruits and vegetables, usually resulting in negative effects on colour,
taste, flavour, and nutritional value. These enzymatic browning also can be
observe in seafood such as shrimps, spiny lobsters and crabs. Enzymatic
browning is the second largest cause of quality loss in fruits and
vegetables. Enzymatic required three conditions that are substrate, enzyme
and oxygen. The reaction is a consequence of phenolic compounds
oxidation by polyphenol oxidase (PPO), which triggers the generation of
dark pigments.
Polyphenols are the main components in enzymatic browning.
Polyphenol are also called phenolic compounds are group of chemical
substances present in plants that play an important role during enzymatic
browning,

because

they

are

substrates

for

the

browning-enzymes.

Polyphenols can be divided into many different sub categories, such as

anthocyanins (colours in fruits), flavonoids (catechins, tannins in tea and
wine) and non-flavonoids components (gallic acid in tea leaves). Flavonoids
are formed in plants from the aromatic amino acids phenylalanine and
tyrosine
Enzymatic browning can be desirable in production of black tea from
green tea leaves, colour development in cocoa, development of golden
brown colour in dried fruits such as raisins, prunes, dates and figs. Although
enzymatic browning is beneficial to the colour and flavour development of
certain food items such as tea, coffee and cocoa, it impairs the quality and
stability of fresh-cut produce. In this experiment, the reaction of enzymatic

browning in several treatments and also the evaluation on colour, odour

and texture prior to enzymatic browning was determined.
MATERIALS
Apples, 1% catechol solution, 1% syrup solution, 20% syrup solution, 1%
citric acid solution, 1% vinegar solution, sugar, water, lemon juice, 1% salt
solution, 1% ascorbic acid solution and 10 ppm sodium metabisulphite
PROCEDURE
A. OBSERVATION OF BROWNING REACTION
PART I
1. Apples was cut into 4 part (approximately 1 quarter each)
2. 1 quarter was reserved intact
3. 1 quarter was bruised
4. 1 quarter was sliced
5. The times taken for the fruit to turn brown was recorded
PART II
1. Apples was cut into 4 part (approximately 1 quarter each)
2. 1 quarter was placed in water
3. 1 quarter was blanched in boiling water for 30 seconds
4. 1 quarter was placed in 1% catechol solution
5. The times taken for the fruit to turn brown was recorded
B. CONTROL OF BROWNING REACTION
1. Apples was cut into quarters and was sliced as it required
2. The cut fruits was treated by following methods which are water, 1%
syrup solution, 20% syrup solution, sugar, 1% salt solution, 1% vinegar
solution, 1% citric acid solution, lime juice, 1% ascorbic acid solution and
10 ppm sodium metabisulphite solution
3. The times taken for the fruit to turn brown was recorded
4. The fruits was evaluated for acceptability including colour, odour and
texture

RESULTS
A. OBSERVATION OF BROWNING REACTION
PART I
SAMPLE

TIME TAKEN FOR THE FRUIT TO TURN BROWN

(minutes)

Reserve

2.15

Bruise

2.14

Crush

0.51

Slice

7.05

PART II

SAMPLE

TIME TAKEN FOR THE FRUIT TO

TURN BROWN (minutes)

Place in water

33.15

Place in 1% catechol
solution

0.59

Undergo blanching

6.15

B. CONTROL OF BROWNING REACTION

SAMPLE

TIME (MIN)

TEXTURE

Water

33.09

Tender

1% Syrup solution

38.04

Tender

20% Syrup solution

40.13

Crisp

Sugar

42.21

Crisp

1% Salt solution

43.05

Crisp

Apple

1% Vinegar solution

24.41

Tender

Pungent

1% Citric acid solution

21.51

Soft,
tender

Apple

Lime juice

44.52

Crisp

Apple &
Lime

1% Ascorbic acid solution

46.15

Crisp

Apple

10 ppm Sodium
metabisulphite

51.54

Very crisp

Apple

DISCUSSION

ODOUR
Fresh
apple
Fresh
apple
Fresh
apple
Fresh
apple

Through the experiments, the reaction of enzymatic browning in

several treatments and also the evaluation on colour, odour and texture
prior to enzymatic browning was determined. Enzymatic browning occurs
through the reaction of phenolic compounds oxidation by polyphenol
oxidase (PPO), which triggers the generation of dark pigments. Apples are
fruits that rich in polyphenols and highly susceptible to enzymatic
browning.
In the first part (Part I), the browning reaction of several sliced of
apples was observed where the sliced apples was reserved, bruised,
crushed and sliced into smaller in size. When fruits or vegetables are peeled
or cut, enzymes contained in the plant cells are released. In the presence of
oxygen from the air, the enzyme phenolase catalyses one step in the
biochemical conversion of plant phenolic compounds to form brown
pigments known as melanin.
From the results, the time taken for the fruits to turn brown which
indicates browning reaction was determined. Cut apples slices that has
been crushed has the fastest time taken to turn brown (0.51 minutes)
meanwhile apples that has been sliced to smaller in size has the slowest
time taken to turn brown (7.05 minutes). For apples that have been bruised
and reserved, the time taken for the apples to turn brown was 2.14 minutes
and 2.15 minutes respectively. Crushing the apple will cause injury to the
apple tissue that disrupts the arrangement of chemicals within the cell and
allow these chemical to make contact with oxygen more rapidly than the
other method.
For second part (Part II), the cut apples was placed in water, 1%
catechol solution and also undergo blanching for around 30 seconds before
the time taken for the apples to turn brown was observed. For apples cut
that have been placed in water, the time taken for the fruits to turn brown
is 33.15 minutes which is the slowest time taken for the fruits to turn
brown. This is because soaking in water alone will temporarily reduce the

level of browning by restricting the amount of oxygen in contact with the

apple slices.
Meanwhile, fruits that undergo blanching also take longer time to turn
brown which is 6.15 minutes where enzyme in the apple might be
denatured during blanching. According to Javdani et al. (2013), blanching
was used to prevent browning reactions and maintain texture in various
vegetables and fruits. In contrast, apples that have been placed in 1%
catechol solution shows the fastest time taken for the fruits to turn brown
which is 0.59 minutes. This is because catechol solution contains a
chemical which resembles a substance which occurs naturally in fruit which
will accelerate the browning reaction.
For control of browning reaction, the apples cuts was treated with
water, 1% syrup solution, 20% syrup solution, sugar, 1% salt solution, 1%
vinegar solution, 1% citric acid solution, lime juice, 1% ascorbic acid
solution and 10 ppm sodium metabisulphite. The acceptability of the fruits
in terms of it texture and odour was then determined. Cuts of apples that
have the longest time to turn brown with good acceptability is apples that
have been treated with 10 ppm sodium metabisulphite where it takes 51.54
minutes to turn brown. It also has very crisps in texture and also was having
fresh apples odour.
Meanwhile, apples that has been treated with 1% of citric acid
solution shows the fastest time taken to turn brown which is 21.51 minutes
with soft and tender texture and have apples odour. On the hand, sliced cut
apples that has been treated with 1% ascorbic acid solution (46.15
minutes), 1% salt solution (43.05 minutes), sugar (42.21 minutes) and 20%
of syrup solution (40.13 minutes) was also the best control method to slow
down browning reaction as it has longer time taken to turn brown with
better acceptability.
During the experiment, they are some sources of error that occur
which may affect the accuracy of the result. For example, the cuts fruit was
not placed in the treated solution immediately as it is exposed to the

oxygen before the treatments has been started. In order to minimize the
errors, the apples should be treated immediately and correct techniques of
handling experiments should be applied.
CONCLUSION
In conclusion, blanching and soaking in water can reduce rate of browning
besides treating the apples in 1% ascorbic acid, 10 ppm sodium
metabisulphite, sugar, 20% sugar solution can be used to control browning
reaction.

REFERENCES
Ioannou, I. & Ghoul M. (2013). Prevention of enzymatic browning in fruit and
vegetables.

Europen Scientific Journal. 9 (30), 1857-7881.

Javdani, Z., Ghasemnezhad, M. & Zare, S. (2013). A comparison of heat

treatment and

ascorbic acid on controlling enzymatic browning of fresh-

cuts apple fruit. International Journal of Agriculture and Crop Sciences, 5(3),
186-193.
STUDY QUESTIONS
1. List the essential components and conditions that must be present if
browning is to occur in fresh fruits and vegetables
Browning can occur in presence of oxygen, enzyme and substrate. It
occurs in injured tissue that was exposed to air or oxygen. The example
of substrate is phenolic compound and the examples of enzyme are
polyphenol oxidase (PPO). This PPO catalyst oxidation of certain phenolic
compound and form browning in fresh fruits and vegetables.
2. Give 5 examples of fresh fruits that are particularly susceptible to
browning

The examples of fresh fruits that are susceptible to browning include

banana, apple, pear, apricot, mango, and peach.
3. Explain six treatments or procedures that might be followed to retard
browning during food preparation and browning
Firstly, browning can be retard by destruction of enzyme by heat
treatment to denature PPO enzyme. Besides browning can be inhibited
using vacuum packaging because oxygen are not available in these
packaging. Sugar and sugar syrups can be used to retard the diffusion of
oxygen. Acid also can be used such as ascorbic acid, citric acid or malic
acid. This acid will low the pH that will inactivate PPO enzyme. Low pH
can prevent or slow down the browning. Furthermore, the addition of
sulphite can inhibit the browning in fruits as it acts as anti-browning
agent.

Lastly, addition of reducing agent can inhibit browning. For

example L-ascorbic acid was used in production of fruit juice and purees.