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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

INTRODUCTION

In 2008, Lin and Zhao said that processed fruits and vegetables have the

most promising development portion within the nourishment retail industry. Due to

spoilage, various specialists have evaluated that up to 25 to 80% of harvested fresh fruits

and vegetables are lost. Deterioration can occur both chemical and physical, water loss,

enzymatic based and light induced colour changes (e.g. browning), oxidation, loss of

cellular integrity (softening) are example of microbiological process that is affected by

deterioration, even the growth of the microorganisms. Listeria monocytogenes is

psychotropic bacteria which is a food-borne pathogen which is relevant for food safety as

the said bacteria can grow in the cooling chain and species of the family

Enterobacteriaceae like Salmonella enteric or Escherichia coli (Muranyi, 2013). This

results in much economic waste in developed countries and more devastating

consequences in many tropical regions in the world that is why producers put the

chemical preservation into the harvested fruits and vegetables.

Elsevier said in 2008 that the use of chemicals on minimally processed fruits and

vegetables have been on less demand as the search for naturally occurring substances are

much more on the highlight to be used as alternative antimicrobials. There are several

preservation methodologies that have been presented. One of these methods of extending

postharvest shelf life is the use of the edible coatings.

In the past years, edible films and coatings have gotten large attention because of

their edges including use as edible packaging materials over synthetic or artificial films

as acknowledged by Bourtoom in 2018. It is applied to multiple products to manage


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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

moisture transfer, gas exchange, or oxidation processes. Edible coatings are also an

environmentally friendly technology and can generate the same effect as modified

atmosphere storage in modifying internal gas composition and it can also give an addition

protective coating to produce (Dhall,2012). They are applied directly on the food exterior

by dipping, spraying or brushing. Edible coatings are used to diminish weight loss during

transport and storage and they are also used to produce a modified atmosphere.

Chitosan is a modified, natural nontoxicbiopolymer derived by deacetylation of

chitin (poly- β-(1→4)-N-acetyl-D-glucose-amine) and it is the second most abundant

polymer. (Maghsoudlou et al.,2012; Sitanggang et al., 2012). It is a kind of fiber which is

from chitin. Chitin is a substance that can be found in the hard-outer shells of crustaceans

such as crab, crayfish, shrimp, and lobster. This is where you can find the chitosan which

is a type of a fiber. For effective control of fruit decay, chitosan can be used for it has

strong antimicrobial and antifungal activities (Aider,2010). According to Abdou in the

year 2013, chitosan could easily form coating on fruit and vegetable, and it can lessen the

respiration rate of fruit and vegetable by adjusting the permeability of carbon dioxide and

oxygen. Though chitosan coating has a lot of edge on the preservation of fruit and

vegetable, it demonstrates a certain defect on that specific fruit or vegetable. One of the

defects is that limited inhibition to special microorganisms which can make the fruit

decay. Another defect on the use of chitosan is the poor coating structure in order to

adjust the permeability of carbon dioxide and oxygen (Jianglian and Shaoying,2013). The

chitosan was mixed with other substances so that it can be effectively applied as an edible

coating (Castell et. al,2013).


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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Essential oils are natural antioxidant and antimicrobial lipidic substances

extracted from plants. Most of them consist of a mixture of terpenes, terpenoids and other

aromatic and aliphatic constituents (Bakalli, 2008). It is said that there has been a large

amount of interest due to the comparatively secured status, general acceptance by buyers

and the utilization for having several benefits which are functional (Chiralt et al., 2012).

By the use of machinery, essential oils are attained by the process of heating and cooling,

though they still produce the same scent and taste because of where they were originated

from. Every essential oil is rarely remarkable, it varies from different constituents of

nature's ingredients that may result from a different smell and absorption on the skin. The

properties of essential oil can have the same general class depending on what

composition is inside of a plant, it can be from plant to plant or the species of that plant.

When it comes to food trade, essential oils are mostly used to make the taste of the food

agreeable, they also acquire a natural antimicrobial that keeps the food fresh and to avoid

any microorganisms. It is important to understand the substances of essentials oils and

how to put or make use it for food preservations. Examples of these are the minimum

inhibitory concentration (MIC) the range of target organisms, the mode of action, and the

effect of food matrix components on their antimicrobial properties.

One essential oil came from Citrus bergamia also known as “Bergamot”; it is a

plant that belongs to the Rutaceae family, defined as a hybrid of bitter orange and lemon.

It is an endemic plant of the Calabria region (Italy). This fruit which is widely known as

Bergamot is mainly being used for the extraction of its essential oil which is commonly

used in perfume, cosmetics, food and confections (Calapai, 2015). It has been reported

that Bergamot Essential Oil (BEO) has both antibacterial and antifungal activity
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

against Campylobacter jejuni, Escherichia coli O157:H7, Listeria monocytogenes,

Bacillus cereus, Staphylococcus aureus and dermatophytes, respectively (Karaca et al.,

2007). The in vitro activity of BEO against Candida spp suggests BEO’s potential role in

the topical treatment of Candida infections (Romano et al., 2005). BEO is also active

against dermatophytes in vitro (Sanguinetti et al., 2007).Bergamot essential oil is as well

used on aromatic ingredient in cosmetics that may be applied on sun exposed skin areas,

but there are also components such as bergapten, citropten, bergamotene, and other

furocoumarins may cause phototoxic effects.

Radish (Raphanus sativus) is broadly developed everywhere throughout the world

and has been utilized for over 1,500 years. The root vegetable known as radish belongs to

the Brassicaceae Family that is consumed all throughout the world and is considered as a

part of the human diet. Radishes have diverse varieties of skin hues, while its substance is

ordinarily white. Likewise, the flavor, size, and length of the radish have different

variations throughout the world. Roots, seeds and leaves are some of the distinctive parts

of radish that is used for restorative purposes. Radish has been utilized ethically as a

laxative, stimulant, biological process aid, appetizer, and within the treatment of

abdomen disorders (Kapoor, 2000).

Throughout the pre-harvest and postharvest stage of the radish it’s physical and

physiological and pathological factors comes it to spoil. Perishing of radish usually occur

since it is a root crop that naturally prefers warm climate. Spring and autumn are the ideal

time for the crop to grow and at the same time it can also tolerates cold climate. The

radishes tend to have tiny tops and roots with strong smell and soft texture when it grows
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

during summer due to the hot temperature. Thus, production of radishes with good

quality during mid-summer can be tough.

Since radishes are extremely helpless to shrivelling, we need to gather them in the

cool and keep them cool and damp until the point when they can be put into cool storage

at a temperature of 0°C and a relative stickiness of 90%. Yet the issue is that not all spots

are appropriate for the radish storage that is the reason makers and customers are

concerned that the collected radish plant will most likely be unable to keep going

sufficiently long for them to be eaten.

Consumers around the world demand for food that is in good quality, without

chemical preservatives, and an extended shelf life. Therefore, an increased effort has

been made to develop new natural preservatives and antimicrobials (Lin & Zhao, 2007).

Different preservation methodologies have been developed. One method of extending

postharvest shelf life is the use of the protective coatings. In analysis of the fact that

consumers demand for natural preservatives is increasing and there is no recent report on

the utilization of Chitosan protective coating with Bergamot Essential Oil in prolonging

the shelf-life and preserving quality of fruits and vegetable.

The overall objective of the researchers is to determine whether chitosan edible

coating with the help of Bergamot Essential Oil is an effective protective coating in

preserving quality and prolonging postharvest physiology of Raphanus sativus. The

researchers’ goal is also to introduce an alternative method which can preserve the

quality and shelf life of fruits and vegetable. Mainly the researchers would like to answer

the following questions;


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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

1. Will the protective coating delay the change of the radish color compared to

the uncoated radish?

2. Will the protective coating delay preserve the usual smell radish compared

to the uncoated radish?

3. Will the protective coating delay the appearance of mold in the radish

compared to the uncoated radish?

4. Will the protective coating be as effective as the commercial gelling agent?

The findings of this study will be a benefit of the society considering that this

research is about having Chitosan with Bergamot Oil as an edible coating in preserving

quality and prolonging the post-harvest physiology of Raphanus sativus. Quality

assurance and shelf-life assurance are both important factors when consumers choose the

food to utilize. The discovery of this research will be beneficial to all. It can specifically

help the producers, vendors and consumers in identifying the processes that can

contribute greatly to these crops by having an alternative way of maintaining its quality

and extending its postharvest physiology. There are crops which usually rot, earlier than

expected due to incompatible temperature and crops are often left outside the

refrigerators. The outcome can also be used as a reference for future researchers who aim

to grasp more on the effectiveness of Chitosan as a protective coating.

This study was conducted at Angeles University Foundation from the month of

August until March 2018-2019. It mainly identified and assessed the effectiveness of

chitosan-based gel with Bergamot Essential Oil on one specific vegetable which is a

radish. Fresh cut radishes with similar weight will be used to acquire more precise data.

The researchers stored the treated and untreated radishes inside a room temperature with
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

a relevant degree of 25- 30-degree celsius. Only shrimp wastes were utilized to create the

chitosan. The study was conducted with limited amount of financial resources since the

researchers are only students. The results of this study can be inferred in different ways,

and the researcher think that there will be no significant differences in the use of Chitosan

edible coating with Bergamot Essential Oil in preserving the quality and prolonging the

shelf-life of Rapahnus savitus.

The method that the researchers used is the method that was introduced by

Iqbal&Hossain in 2015. There are three significant procedures that researchers did during

this experimentation and these are the processes called Demineralization,

Deproteinization and Deacetylation. The researchers used these methods to acquire an

assurance that the experimentation would be successful since the method was already

proven by another researcher.

This study is an example of an economic and biodegradable natural preservative,

applied as a protective coating, on the change of physiochemical parameters and shelf life

in Radish, related to fruit quality during storage.


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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

METHOD

The researchers' used a mixed method procedure in regards of classification of the

research method that was used on this paper. Qualitative and Quantitative research was

used on this paper – description and the narrative style that was used to present the

methodologies, relevant information in the research showed the used of Qualitative

research. Quantitative research method was used in regards of utilization of an

experimental design type of research. This design means to see the effectiveness of

chitosan with bergamot oil as an edible coating to the radish. The method that was

utilized by the researchers was the method used by Iqbal&Hossain in 2015.

Demineralization, Deproteinization and Deacetylation are the most significant procedure

in the method.

In this research, the variables that were used were Chitosan, Bergamot Essential

Oil and radishes.

Chitosan is a sugar that is gotten from the hard-external skeleton of shellfish,

including crab, lobster, and shrimp. It was utilized for prescription and used to treat

obesity, high cholesterol, and Crohn's sickness. It is additionally used to treat

inconveniences that kidney failure patients on dialysis often confront, including high

cholesterol, "tired blood" (iron deficiency), loss of solidarity and craving, and trouble

sleeping (insomnia). Some inquire about recommends that applying chitosan ascorbate

specifically to the gums appears to help in the treatment of periodontitis. Researchers

propose that applying N-carboxybutyl chitosan straightforwardly to the influenced

territory appears to encourage wound recuperating and decrease scar arrangement after
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

plastic surgery. It was likewise referenced that taking chitosan by mouth may diminish

elevated cholesterol, help to address frailty, and enhance physical quality, hunger, and

rest in individuals with kidney disappointment who are accepting long haul hemodialysis.

Some individuals apply chitosan straightforwardly to their gums to treat aggravation that

can lead to tooth loss (periodontitis), or bite gum that contains chitosan to forestall

"holes" (dental caries).

Bergamot is a citrus fruit whose rind was employed for extracting bergamot

essential oil. The scientific name of bergamot is Citrus aurantium var. or Citrus

bergamia. It is a tropical plant however thrives in Europe yet. It is derived from cold

compression, as hostile the steam distillation through that several alternative essential oils

are derived. It helps mitigated depression since its elements like alpha-pinene and

limonene, are antidepressant and stimulating in nature. It helps to stimulate hormonal

secretions and thus helps to maintain correct metabolism. This helps to digest, correctly

absorb nutrients, assimilate and decompose sugar and thus reduce blood sugar. Research

conducted by the Department of Drug Sciences and Health Products of the University of

Messina, Italy, suggests that certain components of the essential bergamot oil are

antibiotic in nature and disinfectant (Delbo et al., 2015). According to research done in

2007, Bergamot essential oil is capable of inhibiting germ, virus and fungus growth. A

research that was conducted at the Department of Pharmacobiology and University

Centre for Adaptive Disorders and Headache, Section of Neuropharmacology of Normal

and Pathological Neuronal Plasticity, University of Calabria, concluded that essential

bergamot oil helps to reduce pain in the body. It stimulates the secretion of certain

hormones that reduce nerve pain sensitivity. It is therefore very useful for headaches,
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

sprains, muscle aches or any other symptoms/diseases that require a high dose of

analgesic pills. The essential oil of Bergamot is a tonic that tones the respiratory,

circulatory, digestive, excretory, and nervous system, as well as the skin and muscles. In

2009, Amantea et al. said it is also anti-congestive and is used in vaporizers to alleviate

congestion and breathing problems, especially in coughs and colds.

Raphanus savitus (Radish) is a root vegetable which was first cultivated in China

years ago that has been able to pave its way to other parts of the world. It is a root

vegetable that has a lot of beneficiaries that can help your body to be healthy. Radishes

are a good source of vitamin C which is an antioxidant that prevents the damaging of

cells caused by aging, an unhealthy lifestyle, and environmental toxins and it also fights

free radicals in your body. It contains similar antioxidants found in citrus fruits and

vegetables (Akruti, 2016). These antioxidants defend the body toward oxidation reactions

which may be cancerous to the body. According to the Higdon (2005), cruciferous

vegetables contain compounds that are broken down into isothiocyanates when combined

with water. Isothiocyanates help purge the body of cancer-causing substances and

prevent tumor development. It is also proven that radish is a natural antifungal which

contains RsAFP2 that can kill a common fungus found in the human body called Candida

albicans (Bink, 2016).

Along with these three main components that will be used in the researchers’

experiment, several chemicals will be used in the experiment. First on the list is Sodium

Hydroxide which is a dangerous and strongly alkaline and was utilized to make synthetic

substances and soap to refine petroleum. Chemical Equation; NaOH. A strong, caustic

alkali NaOH, utilized in the manufacture of soap, paper, detergent, and having other
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

numerous mechanical applications. Glycerol, a dull, sweet, thick fluid formed as a result

in soap manufacture. It was utilized as an emollient and laxative and for making

explosives and antifreeze. Acetic acid is additionally utilized in the usage or production

of pharmaceuticals. In the food industry, acetic acid in lower focus was utilized as

nourishment added to its substance, seasoning and preservative. Acetic acid regulates

food acidity. Lastly, Hydrochloric acid is likewise utilized in the creation of batteries,

photoflash bulbs, and firecrackers. The said chemical is also called as the “workhorse”

chemical for being useful in various ways, proving so, Hydrochloric acid is even used in

processing sugar and making gelatine.

The study was conducted at the laboratory of Angeles University Foundation,

Pampanga. The radishes and the shrimp that was utilized came from a local market in

Pampang, Angeles City, Pampanga. The researchers monitored the experiment for seven

days and that determined the differences that occurred during the experiment.

Materials

The materials that were utilized by the researchers in completing the study differs

from each other depending on what they are supposed to be used for.

First, the researchers prepared the vegetable that was utilized during the

experimentation which is Raphanus sativus (radish). The researchers bought three

radishes that were cut with same shape and the same weight for accuracy that served as

the experimental variable in the study that determined the effectiveness of Chitosan with

edible coating with Bergamot Essential Oil.


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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Second, for the preparation of the Chitosan there were a lot of materials to be

considered. The most important material for the preparation of Chitosan is definitely the

shrimp. The shrimp waste served as an origin of the chitin that will eventually become

chitosan. Tap water was used to wash the shrimp waste to remove all unwanted

contaminants. The Woven Traywas used for the drying of shrimp waste. Mortar and

pestle were used to crush the shrimp waste. Next is polythene bags, this was used to store

the crushed shrimps for the shrimp partial extraction. Strainer was used as a tool for

removing the solvent after demineralization, deproteinization and deacetylation process.

Stirring Rod was used to stir the mixtures. The tool that the researchers will be using to

measure solute and solvents needed are measuring cup, graduated cylinder and weighing

scale. Hydrochloric Acid was used to demineralize the chitin flakes. During the

deproteinization, Sodium Hydroxide was utilized together with distilled water to be able

to make a NaOH Solution. Lastly, essential oil was used to add efficiency of the chitosan

coating.

For the making of chitosan coating, researchers used acetic acid and glycerol to

add efficiency into the chitosan. As for the experimentation process, the researchers

stored the cut radishes inside the Tupperware.

Procedure

The researchers applied an experimental type of research study that would study

the potency of Chitosan coating with the help of Bergamot Essential Oil in preserving

quality and prolonging shelf life of radish.

1. Collection and preparation of radish for coating


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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Three fresh mature radishes were collected from a local market with uniform size,

shape, color, maturity and without any signs of mechanical damage or fungal decay.

After collecting the radishes, surface preparation of the radishes where the researchers

removed all contaminants that would interfere the proper coating application and to give

a sound clean substrate, suitable for firm bonding by washing them thoroughly. After

washing the radishes, the researchers cut the first radish into six parts with uniform size

and weight. The first six radishes which were cut were the first set of samples. The

second and third radishes were also cut into six equal parts with uniform size and weight

and it served as the second and third set of samples.

2. Preparation of Chitosan with Bergamot Essential Oil (BEO) coating

2.1 Preparation of shrimp waste

The researchers collected shrimp head and shrimp skin from a local market in

Pampang, Angeles. The shrimp wastes were then washed with tap water and were

dried under the sun for two days. After drying the shrimp waste, it was then crushed

into small substances which were kept in a polyethylene bags at an ambient

temperature (30±2oC) for 24 hours for partial autolysis that facilitated chemical

extraction of chitosan and to improve the quality of shrimp.

2.2 Isolation of Chitosan

Isolation of chitosan was carried out using the following 3 (three) steps, namely

Demineralization, Deproteinization and Deacetylation.

Demineralization of shrimp shell has been carried out with the use of 3% HCI at

ambient temperature (28±2oC) with a solid to solvent ratio 1:5 (w/v) for 16 hours.
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

After 16 hours, the residue was then washed and soaked in tap water until neutral pH

by the use of pH-Fix.

Deproteinization of shrimp shell was done with 4% NaOH at ambient temperature

(28±2oC) with a solid to solvent ratio 1:5 (w/v) for 20 hours. After 20 hours, the

residue was washed and soaked in tap water until neutral pH. Then purified chitin

was dried until it was become crispy. Chitin flakes was grounded to small particle to

facilitate deacetylation (Removal of acetyl groups from chitin).

Deacetylation was experimented the concentration of NaOH (60%) at 65C

temperature with a solid to solvent ratio 1:10 (w/v) for 20 hours. After 20 hours, the

residue was washed until neutral pH with tap water. The resulting chitosan was then

dried for two hours at 65±50C and Bergamot Essential Oil will be added

subsequently used for coating purposes.

2.3 Making of the Chitosan with BEO coating solution

Sixsolutions were prepared by adding of 1 gram of chitosan powder in a 200 ml

distilled water. The solution resulted to a 0.5% chitosan solution. The researchers

added 0.5% of acetic acid and 0.5 ml of glycerol (0.25% of the weight in grams of

chitosan powder)into the solution to be able to dissolve the chitosan. Bergamot

Essential Oil (0.25%, 0.5% and 1%) was added into three solutions.

3. Application of the Chitosan with BEO coating

In this procedure, three sets of samples containing six radishes were utilized

during this procedure. The chitosan edible coating that was made was applied to the 15

radishes which were cut while the three remaining cut radishes were coated. The

radishes that were coated were dipped into the coating for atleast 15 seconds. The coated
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

and uncoated radishes will be stored into a tupperware. The container of the radishes will

be stored in an ambient storage with a similar temperature.

4. Evaluation of the chitosan protective coating to the radish

In this final procedure, the researchers observed the coated and the uncoated

radishes by its appearance, weight loss, firmness, and its smell. The researchers were able

to determine the difference between the coated and uncoated radishes. This procedure

proved whether the chitosan with bergamot essential oil coating is a really effective

protective coating in preserving the quality and prolonging the shelf life of fruits and

vegetable such as radishes.

To evaluate the weight loss of the radishes, the weight of the radish on the day of

observation was subtracted from its initial weight


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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

RESULTS

The data that were collected by the researchers pertaining to the effectiveness of the

Chitosan with Bergamot Oil based coating in preserving the quality and prolonging the

shelf life were interpreted. The following parameters were observed:

a) Color

Table 1. Color of Coated Radish (Only Chitosan)

Day Sample no.

1 2 3
1 white white white
2 white white white
3 dirty white dirty white dirty white
4 dark brown dirty white dirty white
5 dark brown dirty white dirty white
6 dark brown dirty white dirty white
7 black dirty white light yellow
8 black dark yellow dark yellow

Table 1 presents that the use of Chitosan coating enables the radish to preserve its normal

white color for two days.

Table 2. Color of Coated Radish (Chitosan with 0.25% Bergamot Oil)

Day Sample no.

1 2 3
1 white white white
2 white white white
3 dirty white dirty white dirty white
4 brown dirty white dirty white
5 brown dirty white dirty white
6 brown light yellow dirty white
7 brown light yellow light yellow
8 brown brown yellow
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Table 2 presents that the use of Chitosan with 0.25% Bergamot Oil enables the radish to

preserve its normal white color for two days.

Table 3. Color of Coated Radish (Chitosan with 0.5% Bergamot Oil)

Day Sample no.

1 2 3
1 white white white
2 white white white
3 dirty white dirty white dirty white
4 dirty white dirty white dirty white
5 dirty white dirty white dirty white
6 dirty white dirty white dirty white
7 yellow light yellow dirty white
8 black yellow dark yellow

Table 3 presents that the use of Chitosan with 0.5% Bergamot Oil enables the radish to

preserve its normal white color for two days.

Table 4. Color of Coated Radish (Chitosan with 1% Bergamot Oil)

Day Sample no.

1 2 3
1 white white white
2 white white white
3 dirty white dirty white dirty white
4 dirty white dirty white dirty white
5 dirty white light yellow dirty white
6 dirty white light yellow dirty white
7 dirty white light yellow light yellow
8 yellow yellow yellow

Table 4 presents that the use of Chitosan with 1% Bergamot Oil enables the radish to

preserve its normal white color for two days.


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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Table 5. Color of Coated Radish (Commercial gelling agent)

Day Sample no.

1 2 3
1 white white white
2 white white white
3 yellow dirty white dirty white
4 yellow light yellow dirty white
5 yellow light yellow dirty white
6 yellow light yellow dirty white
7 yellow light yellow dirty white
8 brown brown dirty white

Table 5 presents that the use of commercial gelling agent enables the radish to preserve

its normal white color for two days.

Table 6. Color of Uncoated Radish

Day Sample no.

1 2 3
1 white white white
2 light yellow dirty white dirty white
3 brown dirty white dirty white
4 dark brown yellow light yellow
5 black yellow light yellow
6 black yellow yellow
7 black dark yellow dark yellow
8 black dark yellow dark yellow

Table 6 presents that the untreated radishes shows change of color after one day of

storage, unlike the coated radishes, the uncoated radises was not able to maintain its color

.
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

b) Smell

Table 7. Smell of Coated Radish (Only Chitosan)

Day Sample no.

1 2 3
1 usual smell usual smell usual smell
2 usual smell usual smell usual smell
3 usual smell usual smell usual smell
4 usual smell usual smell usual smell
5 unpleasant unpleasant unpleasant
6 unpleasant unpleasant unpleasant
7 unpleasant unpleasant unpleasant
8 unpleasant unpleasant unpleasant

Table 7 presents the smell of radishes with Chitosan coating applied to it. It shows that

the samples that were tested had a usual smell for four days, but the radishes started to

have an unpleasant odor on its fifth day.

Table 8. Smell of Coated Radish (Chitosan with 0.25% Bergamot Oil)

Day Sample no.

1 2 3
1 usual smell usual smell usual smell
2 usual smell usual smell usual smell
3 usual smell usual smell usual smell
4 usual smell usual smell usual smell
5 usual smell unpleasant usual smell
6 unpleasant unpleasant unpleasant
7 unpleasant unpleasant unpleasant
8 unpleasant unpleasant unpleasant

Table 8 presents the smell of radishes with Chitosan coating applied to it. It shows that

the two of the samples that were tested had a usual smell within five days while the
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

second sample started to grow an unpleasant odor on its fifth day. On the sixth day, the

first and third sample acquired an unpleasant odor.

Table 9. Smell of Coated Radish (Chitosan with 0.5% Bergamot Oil)

Day Sample no.

1 2 3
1 usual smell usual smell usual smell
2 usual smell usual smell usual smell
3 usual smell usual smell usual smell
4 usual smell usual smell usual smell
5 usual smell usual smell usual smell
6 usual smell usual smell usual smell
7 unpleasant unpleasant unpleasant
8 unpleasant unpleasant unpleasant

Table 9 presents the smell of radishes that has a Chitosan with 0.5% Bergamot Oil

coating applied to it. It shows that the samples that were tested had a usual smell within

six day, but the radishes started to grow an unpleasant odor on its seventh day.

Table 10. Smell of Coated Radish (Chitosan with 1% Bergamot Oil)

Day Sample no.

1 2 3
1 usual smell usual smell usual smell
2. usual smell usual smell usual smell
3 usual smell usual smell usual smell
4 usual smell usual smell usual smell
5 usual smell usual smell usual smell
6 usual smell usual smell usual smell
7 unpleasant unpleasant unpleasant
8 unpleasant unpleasant unpleasant

Table 10 presents the smell of radishes that has a Chitosan with 1% Bergamot Oil coating

applied to it. It shows that the samples that were tested had a usual smell within six day,

but the radishes started to grow an unpleasant odor on its seventh day.
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Table 11. Smell of Coated Radish (with Commercial Gelling Agent)

Day Sample no.

1 2 3
1 usual smell usual smell usual smell
2. usual smell usual smell usual smell
3 usual smell usual smell usual smell
4 usual smell usual smell usual smell
5 usual smell usual smell usual smell
6 usual smell usual smell usual smell
7 usual smell usual smell usual smell
8 unpleasant unpleasant unpleasant

Table 11 presents the smell of radishes that has a Chitosan with 1% Bergamot Oil coating

applied to it. It shows that the samples that were tested had a usual smell within seven

day, but the radishes started to grow an unpleasant odor on its eight day.

Table 12. Smell of Uncoated Radish

Day Sample no.

1 2 3
1 usual smell usual smell usual smell
2. usual smell usual smell usual smell
3 usual smell usual smell usual smell
4 unpleasant unpleasant unpleasant
5 unpleasant unpleasant unpleasant
6 unpleasant unpleasant unpleasant
7 unpleasant unpleasant unpleasant
8 unpleasant unpleasant unpleasant

Table 12 presents the smell of the uncoated radishes. It shows that the samples were able

to maintain a usual smell for three days and on its fourth day the samples acquired an

unpleasant smell.
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CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

c) Appearance (Presence of molds)

Table 13.Appearance of Coated Radish ( Only Chitosan )

Day Sample no.

1 2 3
1 none none none
2 none none none
3 none none none
4 none none none
5 present present none
6 present present presemt
7 present present presemt
8 present present presemt

Table 13 presents that the use of Chitosan coating prevented the presence of molds into

the the first and second sample for four days while it prevented the presence of molds

into the third sample for five days . The molds started to appear only on the first and

second sample on its fifth day and only appeared on the third sample on its sixth day.

Table 14. Appearance of Coated Radish (Chitosan with 0.25% Bergamot Oil )

Day Sample no.

1 2 3
1 none none none
2 none none none
3 none none none
4 none none none
5 present none present
6 present present present
7 present present present
8 present present present
Table 14 presents that the use of Chitosan with 0.25% Bergamot Oil coating prevented

the presence of molds into the first and third sample for four days while it prevented the
23
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

presence of molds into the second sample for five days. The molds started to appear only

on the first and third sample on its fifth day and only appeared on the second sample on

its sixth day.

Table 15. Appearance of Coated Radish (Chitosan with 0.5% Bergamot Oil )

Day Sample no.

1 2 3
1 none none none
2 none none none
3 none none none
4 none none none
5 none none none
6 present none present
7 present present present
8 present present present

Table 15 presents that the use of Chitosan with 0.5% Bergamot Oil coating was able tp

prevent the presence of molds into the first and third sample for five days while it

prevented the presence of molds into the second sample for six days. The molds started to

appear only on the first and third sample on its sixth day and only appeared on the second

sample on its seventh day.

Table 16. Appearance of Coated Radish (Chitosan with 1% Bergamot Oil )

Day Sample no.

1 2 3
1 none none none
2 none none none
3 none none none
4 none none none
5 none none none
6 none none none
7 none none none
8 none none none
24
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Table 16 presents that the use of Chitosan with 1 % Bergamot Oil coating prevented the

presence of molds into the samples for seven days.

Table 17. Appearance of Coated Radish (with Commercial Gelling Agent )


Day Sample no.

1 2 3
1 none none none
2 none none none
3 none none none
4 none none none
5 none none none
6 none none none
7 none none none
8 none none none

Table 17 presents that the use of commercial gelling agent prevented the presence of
molds into the samples for seven days

Table 18. Appearance of Uncoated Radish

Day Sample no.

1 2 3
1 none none none
2 present none none
3 present present present
4 present present present
5 present present present
6 present present present
7 present present present
8 present present present

Table 18 presents that uncoated radish prevented the presence of molds into the the first

sample for only one day and while it prevented the presence of molds into the second and

third sample for two days. The molds started to appear only on the first on its second day

and only appeared on the second and third sample on its third day.
25
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

d) Weight Loss

Table 19. Weight Loss of Treated and Untreated Radish in grams

Treatment Sample no. Days

1 2 3 4 5 6 7 8
Untreated 1 14 2(12) 4(10) 6(8) 7(7) 8(6) 9(5) 12(2)
2 14 3(11) 3(11) 5(9) 6(8) 7(7) 8(6) 11(3)
3 14 3(13) 3(12) 5(9) 7(7) 7(7) 8(6) 12(2)
Chitosan 1 14 3(13) 3(11) 4(10) 6(8) 8(6) 9(5) 9(5)
2 14 2(12) 2(12) 2(12) 6(8) 7(7) 8(6) 8(6)
3 14 1(13) 1(13) 3(11) 5(9) 6(8) 6(8) 7(7)
Chitosan w/ 1 14 1(13) 1(13) 2(12) 3(11) 4(10) 4(10) 5(9)
0.25% BEO 2 14 1(13) 2(12) 2(12) 2(12) 5(9) 5(9) 6(8)
3 14 1(13) 1(13) 2(12) 3(11) 4(10) 5(9) 6(8)
Chitosan w/ 1 14 0(14) 1(13) 2(12) 2(12) 7(7) 12(2) 12(2)
0.5% BEO 2 14 0(14) 0(14) 2(12) 3(11) 4(10) 7(7) 10(4)
3 14 0(14) 1(13) 2(12) 2(12) 4(10) 6(8) 8(6)
Chitosan w/ 1 14 1(13) 1(13) 1(13) 2(12) 2(12) 3(11) 4(10)
1% BEO 2 14 0(14) 1(13) 1(13) 1(13) 2(12) 4(10) 6(8)
3 14 0(14) 0(14) 0(14) 1(13) 2(12) 3(11) 6(8)
Commercial 1 14 0(14) 1(13) 2(12) 3(11) 4(10) 4(10) 4(10)
Gelling Agent 2 14 0(14) 1(13) 2(12) 2(12) 3(11) 3(11) 6(8)
3 14 1(13) 1(13) 2(12) 3(11) 2(12) 3(11) 5(9)

Table 19 presents the weight loss in grams per day of each coated and uncoated

variables. For the last day of storage, the weight loss in grams of the uncoated variable

group is greater than the grams of the coated samples.


26
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Statistical Treatment

The researchers used Analysis of Variance or also known as ANOVA Testing in

order to see if the experiment results are significant. It is a statistical treatment where the

researchers were able to analyze the differences among group means in a sample. If the

p-value of the result in ANOVA Testing is significant, it means that the researchers must

use T-test with two-sample assuming unequal means to identify which of the treatments

will have a difference. To be able to identify if there is a significant difference between

the treatments p-value should be less than 0.05.

Table 20 ANOVA Test Results for weight loss between the positive control and the

SS Df MS F P-value F crit
55.06666667 4 13.76666667 8.97826087 0.002407067 3.47805
15.33333333 10 1.533333333

70.4 14

coatings

Table 20 presents that the p-value 0.002407067 of the results for weight loss between the

positive control and the edible coatings have a significant difference.

Table 21. T-test Result for weight loss between the radishes with commercial gelling

agent and radishes with Chitosan coating

commercial gelling agent Chitosan coating


Mean 5 8
Variance 1 1
Observations 3 3
Df 4
P(T<=t) two-tail 0.021311641
27
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Table 21 presents that the p-value 0.021311641 of the results for weight loss between the

radishes with commercial gelling agent and the radishes with chitosan coating have a

significant difference. We can interpret that Chitosan coating cannot be as effective as the

positive control which is the commercial gelling agent.

Table 22. T-test Result for weight loss between the radishes with commercial gelling

agent and radishes with Chitosan and 0.25% of Bergamot Oil coating

Chitosan with 0.25%


commercial gelling agent Bergamot Oil coating
Mean 5 5.66666666666667
Variance 1 0.333333333333336
Observations 3 3
Df 3
P(T<=t) two-tail 0.391002218469803

Table 22 presents that the p-value 0.0.391002218469803 of the results for weight loss

between the radishes with commercial gelling agent and the radishes with with 0.25%

Bergamot Oil coating have no significant difference. We can interpret that Chitosan with

0.25% Bergamot Oil coating can be as effective as the positive control which is the

commercial gelling agent.

Table 23. T-test Result for weight loss between the radishes with commercial gelling

agent and radishes with Chitosan with 0.5% of Bergamot Oil coating

Chitosan with 0.5%


commercial gelling agent Bergamot Oil coating
Mean 5 10
Variance 1 4
Observations 3 3
Df 4
P(T<=t) two-tail 0.0304662916591708
28
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Table 23 presents that the p-value 0.0.391002218469803 of the results for weight loss

between the radishes with commercial gelling agent and the radishes with 0.5%

Bergamot Oil coating have a significant difference. We can interpret that Chitosan with

0.5% Bergamot Oil coating cannot be as effective as the positive control which is the

commercial gelling agent.

Table 24. T-test Result for weight loss between the radishes with commercial gelling

agent and radishes with Chitosan and 0.5% of Bergamot Oil coating

Chitosan with 1%
commercial gelling agent Bergamot Oil coating
Mean 5 5.33333333333333
Variance 1 1.33333333333334
Observations 3 3
Df 4
P(T<=t) two-tail 0.724658636519015

Table 24 presents that the p-value 0.0.391002218469803 of the results for weight loss

between the radishes with commercial gelling agent and the radishes with 1% Bergamot

Oil coating have no significant difference. We can interpret that Chitosan with 1%

Bergamot Oil coating can be as effective as the positive control which is the commercial

gelling agent.

Table 25. ANOVA Test Results for weight loss between the negative control and the

coatings

SS df MS F P-value F crit
50.26666667 4 12.56666667 8.195652174 0.003370759 3.47805
15.33333333 10 1.533333333

65.6 14
29
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Table 25 presents that the p-value 0.003370759 of the results for weight loss between the

positive control and the edible coatings have a significant difference.

Table 26. T-test Result for weight loss between the untreated radishes and radishes with

Chitosan coating

No coating Chitosan coating


Mean 9 8
Variance 1 1
Observations 3 3
Df 4
P(T<=t) two-tail 0.287864134785968

Table 26 presents that the p-value 0.287864134785968 of the results for weight loss

between the radishes with untreated radishes and the radishes with chitosan coating have

no significant difference. We can interpret that Chitosan coating is notan effective

alternative coating in preserving the qualityof radishes.

Table 27. T-test Result for weight loss between the untreated radishes and radishes with

Chitosan and 0.25% Bergamot Oil coating

Chitosan with 0.25%


No coating Bergamot Oil coating
Mean 9 8
Variance 1 1
Observations 3 3
Df 4
P(T<=t) two-tail 0.0153924380719779

Table 27 presents that the p-value 0.0153924380719779 of the results for weight loss

between the radishes with untreated radishes and the radishes with chitosan with 0.25%
30
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Bergamot Oil coating have a significant difference. We can interpret that Chitosan with

0.25% Bergamot Oil coating is an effective alternative coating in preserving the quality

of radishes.

Table 28. T-test Result for weight loss between the untreated radishes and radishes with

Chitosan and 0.5% Bergamot Oil coating

Chitosan with 0.5%


No coating Bergamot Oil coating
Mean 9 10
Variance 1 4
Observations 3 3
Df 4
P(T<=t) two-tail 0.495025346091885

Table 28 presents that the p-value 0.0153924380719779 of the results for weight loss

between the radishes with untreated radishes and the radishes with chitosan with 0.5%

Bergamot Oil coating have no significant difference. We can interpret that Chitosan with

0.5% Bergamot Oil coating is not an effective alternative coating in preserving the

quality of radishes.

Table 29. T-test Result for weight loss between the radishes with untreated radishes and

radishes with Chitosan with 1% Bergamot Oil coating

Chitosan with 0.5%


No coating Bergamot Oil coating
Mean 9 10
Variance 1 4
Observations 3 3
Df 4
P(T<=t) two-tail 0.0141725949711996
31
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

Table 29 presents that the p-value 0.0153924380719779 of the results for weight loss

between the radishes with untreated radishes and the radishes with chitosan with 1%

Bergamot Oil coating have a significant difference. We can interpret that Chitosan with

1% Bergamot Oil coating is an effective alternative coating in preserving the quality of

radishes.
32
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

SUMMARY OF FINDINGS

The statistical treatments that had been utilized by the researchers are ANOVA

Testing and T-test. The ANOVA Testing reveals that there is a significant difference

between the use of the Chitosan with Bergamot Oil coating and the positive control

which is the commercial gelling agent since the p-value 0.002407067 is less than 0.05.

The t-test revealed that the use of Chitosan coating and Chitosan with 0.05% of Bergamot

Oil is not as effective as the commercial gelling agent in preserving the quality of radish

since their p-value is greater than 0.05. While the t-test revealed that the use of Chitosan

with 0.25% of Bergamot Oil and Chitosan with 1% of Bergamot Oil is as effective as the

commercial gelling agent in preserving the quality of radish since their p-value is greater

than 0.05.

The ANOVA Testing also reveals that there is a significant difference between

the use of the Chitosan with Bergamot Oil coating and uncoated radishes since the p-

value 0.003370759 is less than 0.05. The t-test revealed that the use of Chitosan coating

and Chitosan with 0.05% of Bergamot Oil is not an effective edible coating in preserving

the quality of radish since their p-value is greater than 0.05. While the t-test revealed that

the use of Chitosan with 0.25% of Bergamot Oil and Chitosan with 1% of Bergamot Oil

is an effective edible coating in preserving the quality of radish since their p-value is

greater than 0.05.

CONCLUSION

The use of Chitosan coating with Bergamot Essential Oil is effective to preserve

the quality and prolong the shelf-life of different vegetables such as radish. The p-value
33
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

of the ANOVA Test results determined that there is a significant difference between the

coatings and the positive and negative control which are the commercial gelling agent

and untreated radishes subsequently because the p-value from both test are less than 0.05.

Because of that, the researchers proceeded with the t-test of two sample mean with

unequal variances. The researchers were able to identify which of the coating served as a

good alternative preservative in prolonging shelf-life and preserving the quality of

radishes. The results show that Chitosan with 0.25% Bergamot Oil and Chitosan with 1%

Bergamot Oil is an effective coating. Based from the observations of the researchers in

the application of the coating, it prevented the forming of molds and spoiling of radish

later than the uncoated radishes. The radishes with coating also prevented the change of

the color for almost two days. Additionally, the smell of uncoated radishes from the

second day acquired an unpleasant smell compared to the coated radishes which were

able to maintain its usual smell for at least five days.

RECOMMENDATION

Based on the findings and conclusion of the research conducted, here are several

recommendations to be considered:

1. The researchers recommend this study to be used in future reference towards in

assistance of further improvement of agricultural industry regarding crops.

2. The researchers suggest the future researchers to bring the sample in Food and Drug

Administration (FDA) and Department of Health (DOH) for further inspection if it is safe

to eat the experimented edible coating.


34
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

3. The researchers advised the future researchers in considering the limitations that had occurred

during the experiment such as:

a.) Fruits and vegetable that will be used will be harvested on the same day

b.) Use different kinds of essential oils

c.) Use different kinds of crustaceans not only shrimp waste

d.) In testing the samples, use different kinds of method, not only the weight loss
35
CHITOSAN WITH BERGAMOT OIL AS A PROTECTIVE COATING TO RADISH

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