USING HONEY AND WATER TO PREVENT ENZYMATIC BROWNING

Chapter I

Introduction

Background of the Study

Prepping fruits and vegetables ahead of time makes putting together the final dish a lot
easier, but once you cut them up, they start to turn brown almost instantly. However, thanks
to this quick and easy tip from America’s Test Kitchen, you can keep them looking fresh for
much, much longer. Most cooks would tell you to try using lemon water instead, but dunking
fruits and veggies in honey water for 30 seconds prevents them from browning for a lot longer,
even as long as 8 hours. Dunk them for a couple minutes and they could even last a whole day!
The reason this trick works is because a peptide compound found in honey prevents the
enzyme polyphenol oxidase from converting phenolic compounds into dark pigments. Many
people add a little lemon or citrus to keep sliced fruit like apples or pears from browning but it
turns out a mixture of honey and water is much better than adding lemon or citrus fruit to the
fruit and it also keeps your fruit fresher for much longer. Enzymatic browning is one of the
most important reactions that takes place in most fruits and vegetables as well as in
seafood. These processes affect the taste, color, and value of such foods. Generally, it is a
chemical reaction involving polyphenol oxidase, catechol oxidase, and other enzymes that
creates melanins and benzoquinone from natural phenols. Enzymatic browning (also called
oxidation of foods) requires exposure to oxygen. It begins with the oxidation
of phenols by polyphenol oxidase into quinones, whose strong electrophilic state causes high
susceptibility to a nucleophilic attack from other proteins. These 1uinines are then polymerized
in a series of reactions, eventually resulting in the formation of brown pigments (melanosis) on
the surface of the food.The rate of enzymatic browning is reflected by the amount of active
polyphenol oxidases present in the food.Hence most research investigating methods to inhibit
enzymatic browning has focused on hindering polyphenol oxidase activity. However, not all
browning of food produces negative effects.

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Statement of the Problem

A lot of people are complaining about fruits having a rotting color when not eaten. This is
called to a chemical process which is called enzymatic browning. It is caused by an enzyme
called polyphenol oxidase which results in brown pigments when the cells from reaction. So we
planned to use honey and water to prevent enzymatic browning. What will happen? Fruits or
vegetables are exposed to oxygen. Bruising on a fruit is caused by the same kind of problem.

1. Can honey really prevent the enzymatic browning of the fruits and vegetables?
2. About how many tablespoons of honey should we use
3. Are there any other solutions to the problem?
4. Do you think the experiment will work?

Null Hypothesis

H01: The apple will still have a hint of brown

H02: The experiment will not work

Significance of the Study

The importance of the study is to help people in the environment to find a solution to
enzymatic browning for it to stop. It is also for chefs and bakers that can be able to use this
technique in their dishes, desserts and so on.

Scope and Delimitation

The scope of this study is about honey because it helped our apple prevent to have
enzymatic browning. Honey is a sweet, syrupy substance produced by honeybees from the nectar
of flowers and used by humans as a sweetener and a spread. Honey is also helpful in many ways.

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Definition of Terms
To provide clear understanding on the subject of this study, the following terms are
defined and operationally:
Apple: An apple is a sweet, edible fruit produced by an apple tree. Apple trees are cultivated
worldwide and are the most widely grown species in the genus Malus. The tree originated in
Central Asia, where its wild ancestor, Malus sieversii, is still found today.
Honey: Honey is a sweet, viscous food substance produced by bees and some related insects.
Bees produce honey from the sugary secretions of plants or from secretions of other insects, by
regurgitation, enzymatic activity,
Polyphenol Oxidase: Polyphenol oxidase is a tetramer that contains four atoms of copper per
molecule, and binding sites for two aromatic compounds and oxygen. The enzyme catalyses the
o-hydroxylation of monophenol molecules in which the benzene ring contains a single hydroxyl
substituent to o-diphenols.
Enzymatic Browning: Enzymatic browning is the process of food turning brown due to the
chemical reactions that take place within. The process of enzymatic browning is one of the
chemical reactions that take place in food chemistry and represents an interesting research topic
regarding health, nutrition, and food technology.
Melanin: Melanin is a broad term for a group of natural pigments found in most organisms.
Melanin is produced through a multistage chemical process known as melanogenesis, where the
oxidation of the amino acid tyrosine, is followed by polymerization.
Benzoquinone: Benzoquinone, commonly known as para-quinone, is a chemical compound with
the formula. In a pure state, it forms bright-yellow crystals with a characteristic irritating odor,
resembling that of chlorine, bleach, and hot plastic or formaldehyde.
Catechol Oxidase: Catechol oxidase is a copper oxidase that contains a type 3 di-copper
cofactor and catalyzes the oxidation of ortho-diphenols into ortho-quinones coupled with the
reduction of molecular oxygen to water. It is present in a variety of species of plants and fungi
including Ipomoea batatas and Camellia sinensis.

Nucleophilic:(of a molecule or group) having a tendency to donate electrons or react at electron-

poor sites such as protons.

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 Chapter II

Review of Related Literature

This study evaluated the antioxidative capacity of 13 US Northwest honeys from

different floral sources and their anti-browning effect on fresh-cut apples. The inhibitory effect
of honey on enzymatic browning of fresh-cut apples were studied by simply immersing apple
slices in 10% honey solution for 30 min or vacuum impregnating (vacuum at 75 mmHg for 15
min followed with 30 min restoration at atmospheric pressure) in the same honey solution. The
10% diluted high-fructose corn syrup solution was used as a comparison. The surface color of
the apple slices was monitored during 14 days of storage at 3 degrees C and 90% relative
humidity. Physicochemical properties of the apples immediately after treatment were also
evaluated. Wildflower honey had the darkest color and the highest antioxidative capacity among
all test honeys. Vacuum impregnation with honey was more effective in controlling browning
discoloration than that of simple immersion treatment. Honey in combination with vacuum
impregnating operation may have a great potential for developing high-quality fresh-cut fruits.

https://www.ncbi.nlm.nih.gov/pubmed/16009631

Enzymatic browning of fruits and vegetables has not ceased to be a problem for
processors, although the underlying basic reactions have been known for a long time. One of the
difficulties in selecting the mode of browning prevention consists in the necessity of complying
with food safety regulations, while at the same time taking into account the marketability of the
product as affected by taste and flavor, texture, etc. The ever increasing amount of literature
accumulated on the topic shows great progress in establishing the phenolic composition of foods,
enzyme purification and the introduction of new browning inhibitors. This review gives an
account on the latest achievements in the prevention of browning along with some of the
author’s earlier results.

https://pubs.acs.org/doi/abs/10.1021/bk-1995-0600.ch004?src=recsys

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Unlike other sweeteners, honey has shown antioxidant activity that provides this food with
nutritional and technological advantages. Honey has proved to prevent or delay food spoilage
due to oxidative reactions, protecting meats against lipid oxidation (Antony, Rieck, & Dawson,
2000;McKibben & Engeseth, 2002), and vegetable products against enzymatic browning (Chen
et al., 2000;McLellan, Kime, Lee, & Long, 1995;Oszmianski & Lee, 1990). Therefore, honey
has a great potential to be used as a natural antioxidant for foods (Gheldof et al., 2002;Nagai,
Inoue, Kanamori, Suzuki, & Nagashima, 2006).Many of these substances have antioxidant
activity that makes them preservatives (Chen et al. 2000). Honey has been investigated as an
antibrowning agent in raisins (10% and 20% honey solution; McLellan et al. 1995), grape juice
(Lee 1996) and fresh- cut apple (10% honey solution; Jeon and Zhao 2005). Furthermore, honey
prevents enzymatic browning of fruit and vegetable homogenates (Chen et al. 2000), and of
fresh-cut apples (Jeon and Zhao 2005). The first batch of cubes was dipped into 10% diluted
honey solution, the second batch into 20% diluted honey solution, and the third batch into water
(the control), each for 5 min. The 10% diluted solution was chosen due to a previous honey dip
related work (McLellan et al. 1995) and the 20% diluted solution due to similarity with soluble
solids content (SSC) of persimmons used the present study. The cubes were removed from the
water or diluted honey solution using a plastic strainer and then drained by shaking the strainer
that was half-filled with cubes. Ates et al. (2001) reported that a peptide from honey could inhibit
polyphenol oxidase (PPO) extracted from mushroom. Recently, honey has been proved as an
effective anti- browning agent by inhibiting browning activity in raisins ( McLellan et al., 1995),
fresh-cut apple (Oszmianskii and Lee, 1990;Jeon and Zhao, 2005), fresh-cut persimmon ( Son et
al., 2001;Ergun and Ergun, 2010) and fresh-cut mango fruit. This refers to the formation of a
brown color in plant anthocyanin extracts as a result of a two-step process. First, anthocyanins
are oxidized by plant polyphenol oxidases present in the plant extract (Oszmianski and Lee
1990; Mclellan et al. 1995; Tsai et al. 2004). Second, the oxidized anthocyanins undergo
condensation and form brown pigments, which are usually undesired by the food industry.
Honey has been used to inhibit polyphenoloxidase activity in apple slices and grape juice
(Oszmianski & Lee, 1990). Indeed, honey acts as an antibrowning agent in light raisin
processing and results in lighter and more yellow raisins than those fruits produced by the com-
mercial, pressure-infused sulphite solution treatments (McLellan et al., 1995). Surface color
changes can be considered a major sensory parameter and an important attribute of a food

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product such as dehydrated fruit. The best sensory evaluation score (4.03) was recorded for the
berries stored after immersion in honey, while control 1Changes in weight loss (a), soluble solid
content (b), titratable acidity (c) and maturity index (d) of minimally processed grape berries
during storage (n=3) fruits (3.34) declined to the critical level of acceptability, probably due to
higher water loss. In a similar manner, McLellan et al. (1994) evaluated the effects of various
agents (commercial sulfur dioxide gas, honey solution, sulfite, erthorbic acid, citric acid and
CaCl 2) on quality of Thompson Seedless raisin. Among them, honey treatment alone was
ranked the highest by each of the 10 panelists, indicating the promising feature of honey dip
treatment on reducing quality loss. Many of these substances owe their preservative properties to
their antioxidant activity (Chen et al., 2000). Honey has been investigated for browning control
in raisin (McLellan et al., 1995), grape juice (Lee, 1996) and fresh-cut apple (Jeon and Zhao,
2005), for preventing/delaying both spoilage and microorganisms and foodborne pathogens in
different media (Mundo et al., 2004). Thus, honey dip treatment is a potential tool to control
enzymatic browning and to maintain quality of minimally processed pomegranate arils. As
mentioned previously, honey has a wide number of substances that may act in this way,
including ascorbic acid, small peptides, flavonoids, α-tocopherol, and enzymes such as glucose
oxidase, catalase, and peroxidase (Ferreas and others 1993; Jeon and Zhao 2005). Therefore,
honey may be a natural alternative to sulfites for controlling enzymatic browning during fruit and
vegetable processing and for obtaining juices and preserves (McLellan and others 1995; Lee
1996). Parkin et al. (2003) observed that beetroot extracts have strong protecting effect towards
β-carotene. Similar to beet, honey, which has been used as sweetening agent from ancient times
also serve as a good source of natural antioxidants that are effective in preventing oxidative
deterioration in foods (Wang, Gheldof, & Engeseth, 2004) such as inhibiting browning reaction
in fruits and vegetables (Chen, Mehta, Berenbaum, Zangerl, & Engeseth, 2000; McLellan, Kime,
Lee, & Long, 1995; Oszmianski & Lee, 1990) and lipid oxidation in cooked ground poultry
(Antony, Han, Rieck, & Dawson, 2002; Antony, Rieck, & Dawson, 2000; McKibben &
Engeseth, 2002). The combine activity of many components in honey (ascorbic acid, phenolic
compounds, Maillard reaction products, enzymes like peroxidase and catalase etc.) might be
responsible for overall antioxidant property of honey (Wang et al., 2004). Most carboxylic acids
have shown inhibitory effects on enzymatic browning due to their chelating characteristics and
lowering of pH (Son et al., 2001). Since honey showed potent ant browning activity in raisins

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(McLellan et al., 1995) and juices (Lee, 1996; Gacche et al., 2009), it can be use for preventing
browning in fresh-cut fruit. Many previous works had reported that honey solution retarded
browning discoloration in fresh-cut apple (Oszmianski and Lee, 1990; Son et al., 2001; Jeon and
Zhao, 2005) and fresh-cut persimmon (Ergun and Ergun, 2010).

https://www.researchgate.net/publication/229713068_Effect_of_honey_as_an_anti-
browning_agent_in_light_raisin_processing

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 Chapter III

Methodology

Procedure:
In this experiment you will be needing the following items:

 Honey
 Apple
 Knife
 Bowl
 Water
 Spoon

After gathering the materials you should do the following steps:

Step 1: Cut the apple into slices and set them aside

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Step 2: Pour ¼ cup of water to the bowl then add 2 tablespoons of honey to the water

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Step 3: Place the slices into the solution, but leave one out for comparison

Step 4: Compare and contrast both of the apples.

(Left being the one in the solution whileright being the one left out the solution.)

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 Chapter IV

Presentation, Analysis and Interpretation of Data

4 – Outstanding: The apple’s color has no change at all, the apple is very crunchy.
3 – Very Satisfactory: The apple’s color is slightly brown, the apple is slightly soggy.
2 – Satisfactory: The apple’s color is very brown, the apple is soggy.
1 – Needs Improvement: The apple’s color is completely brown, the apple is very soggy.

Characteristics of the apple. Apple in the solution. Apple not in the solution.

The rotting of the apple 43 2 1 4 3 2 1

No enzymatic browning happened. 4 3 2 1 4 3 21

The apple is still crunchy. 4 3 2 1 4 3 2 1

The table above shows the state of the apple after we left it for 24 hours. The grading
goes 4, 3, 2 then 1. 4 means it’s outstanding, the apple has no change at all and it is very crunchy.
3 means very satisfactory, the color of the apple is just slightly brown and it is just slightly
soggy. 2 means satisfactory, the apple has become very brown and soggy. And finally, 1 means
needs improvement, the apple has become completely brown and it is very soggy.

Results

For the first characteristic, the apple in the solution gets a 3 because it became slightly
brown but for the apple not in the solution it gets a 1 because the apple became completely
brown. For the second characteristic, the apple in the solution gets another 3 because again the
apple is slightly brown and for the apple not in the solution, it gets a 1 because the apple has
become completely brown. For the third and final characteristic, the apple in the solution gets a 4
because the apple is still crunchy and for the apple not in the solution gets another 1 because it is
very soggy.

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 Chapter V

Summary, Conclusion and Recommendation

Summary

This experiment is all about how to prevent enzymatic browning from happening. We
decided to choose this experiment because it has become a problem in our community; it has
been affecting the foods that we eat and the foods that we make for ourselves or for others. We
looked for a solution for the problem and we found 2 options, the first one was to use lemon and
the second one was honey, we decided to use honey because honey because it works much better
than lemon and a 30 second dip from the solution can make the fruit last for a solid 8 hours. So
we tested it and it worked! We put sliced apples into a solution of honey and water for 24 hours
and left one slice next to it. The apple in the solution became slightly brown but still crunchy, the
opposite happened for the apple not in the solution. We hope that this can help people and the
community.

Conclusion

Based on the table that we created, the result of our experiment was that it was
successful. It was successful because when we tried our experiment, the experiment worked right
away as we expected.

 Our experiment worked therefore, chefs and bakers may use this technique in their foods.
 The results of the experiments was that the apple’s color was better if it was in the honey
and water solution.
 The experiment showed that the apple’s texture became soggy if it was not in the honey
and water solution.

Recommendation

In consideration to the results of this study the researchers have drawn the following
recommendations:

 Our recommendation is to use honey and water instead of lemon or citrus fruits.

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 We recommend this Science Investigatory Project to other people because fruit and
vegetables rotting is rampant in our society these days.

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 REFERENCES

https://www.wikipedia.com

https://www.google.com

https://www.researchgate.net/publication/229713068_Effect_of_honey_as_an_anti-
browning_agent_in_light_raisin_processing

https:/lifehacker.com

https://pubs.acs.org/doi/abs/10.1021/bk-1995-0600.ch004?src=recsys

https://www.ncbi.nlm.nih.gov/pubmed/16009631

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