EFFECT OF EDIBLE BASED COATINGS INCORPORATED

WITH ALOE VERA ON THE SHELF-LIFE OF FRESH CUT

APPLE

NOR SYAHIRAH BINTI MAHMAD SHAUPI

Written Proposal Submitted in

Partial Fulfillment of the Requirements for the
Degree of Bachelor of Science (Hons.) Biology
In the Faculty of Applied Sciences
Universiti Teknologi MARA

JUNE 2018
 TABLE OF CONTENTS

PAGE

1.0 INTRODUCTION
1.1 Background Study 1
1.2 Problem Statement 3
1.3 Significance of the Study 4
1.4 Objectives of the Study 5

2.0 LITERATURE REVIEW

2.1 Aloe Vera (Aloe barbadensis miller) 7
2.1.1 Chemical composition of aloe vera 7
2.1.2 Aloe vera in context of coating 8
2.1.3 Antimicrobial efficacies of aloe vera 9
2.2 Quality Atrributes 10
2.2.1 Appearance and color 10
2.2.2 Texture 11
2.3 Edible Coating 12
2.3.1 Gelatin films 13
2.3.2 Calcium chloride 14
2.4 Acidifying Agent 15
2.4.1 Ascorbic acid 16
2.4.2 Citric acid 17
2.5 Synthetic Coating on Fruits 17

3.0 METHODOLOGY
3.1 Materials 19
3.1.1 Raw materials 19
3.1.2 Chemicals 19
3.1.3 Apparatus 19
3.2 Methods 20
3.2.1 Sample preparation 20
3.2.2 Preparation of aloe vera gel 20
3.2.3 Preparation of calcium chloride solution 21
3.2.4 Preparation of gelatin solution 21
3.2.5 Coating the Apple Slices 21
3.3 Product Quality Attribute Analysis 22
3.3.1 Measurement of Titrable Acidity (TA) 22
3.3.2 Weight loss determination 23
3.3.3 Sensory evaluation 23
3.3.4 Microbial evaluation 24
3.4 Statistical Analysis 24

CITED REFERENCES
GANTT CHART
STUDENT-SUPERVISOR MEETING FORM
1.0 INTRODUCTION

1.1 Background of Study

Consumer acceptance toward fresh fruit depends on the quality of the fruit.

Appearances of the fruits play a vital role to describe their quality in terms of

size and color uniformity, glossiness, and absence of defects in shape or skin

finish. In addition, quality of high perishable fruit will decrease due to

senescence, oxidation, and microbial growth. Therefore, one method of

extending the postharvest shelf life of the fruit is by the application of edible

coatings. Edible coatings are thin films of consumable material deposited on

the surface of the fruits for additional protective of natural wax (McHugh &

Senesi, 2000).

The use of edible coatings have showed its effectiveness in improving food

attribute and prolonging shelf life of fruits and vegetable by protecting the

products from mechanical and microbial damage (Arnon, Granit, Porat, &

Poverenov, 2015). Moreover, edible coating can avoid water loss, at the same

time preventing shriveling, increasing marketability, and maintaining their

quality through delayed ripening and senescence (Mahajan, Tandon, Kapoor,

& Sidhu, 2017). Usually, shellac and carnauba wax are used by the food

industry as edible coating for apples and citrus fruits. Petroleum based waxes

and resin also being used to coat the fruits (Dhall, 2016)

1
Recently, a number of studies of using Aloe Vera gel-based edible coating

element for fruits and vegetable have been carried out earlier (Misir, Brishti,

& Hoque, 2014; Song et al., 2013). Aloe vera is a cactus-like plant that

belongs to the family Liliaceace. It is widely known that aloe gel have

medicinal and therapeutic properties (Rajeswari et al., 2012). Aloe vera gel

also provide the evidence of the best consumable and biologically safe

preservative films for variety types of fruits and vegetable due to its film-

forming properties, antimicrobial actions, biodegradability and biochemical

process (Misir, Brishti, & Hoque, 2014). Another study proved that aloe vera

incorporated with gelatin can extend the shelf life of fresh cut oranges (Radi,

Firouzi, Akhavan, & Amiri, 2017). The use of gelatin in the study will helps

to reduce oxygen, moisture and oil transport (Bourtoom, 2008).

Hence, for keeping the above views, this study will demonstrates the

effectiveness of Aloe vera as edible coating. Furthermore, this study will

measure the ability of the Aloe vera to prolong the conservation of fresh fruit

since fresh-cut apples are more difficult to preserve than whole apple due to

browning effect.

2
1.2 Problem Statements

Fruits and vegetables are very nutritious and form as essential component in

human diets. This fresh produce had encountered some problems due to their

low shelf life. Furthermore fruits and vegetables are easily susceptible to

physiological decay and quality loss after harvested. Fruits and vegetables

however, can be classified as perishable living products that require more

attention from growers, processors and retailers to maintain quality of the

fruit. Moreover, about 80 % of harvested fresh fruits was estimated to lost due

to spoilage (Arowora et al., 2013). In order to overcome these problems, there

is a need to develop methods to preserve commodities for extra storage time.

Fruits usually subjected with many different environmental effects such as

enzymatic browning and contamination by bacteria. As a result, undesirable

reaction can occur that will cause the fruits to become rotten. When the fruit is

visibly spoiled, it contains large of bacterial colonies growing in it (Gram et

al., 2002). Therefore it will cause adverse health effect if the fruits are

consumed. Thus, the fruits require protection to avoid spoilage during storage.

Traditional preservation techniques such as drying, salting and acidification

have been used to prevent the spoilage and growth of bacteria in fruits, but it

will reduce the nutritional content inside the fruits. In contrast, preservation

by using edible coating has been demonstrated to be more effective in

protecting the fruits without degrading the nutritional value inside the fruit.

3
 The development and application of these aloe vera based edible coating is

limited due to two main factors. First, there is a lack of knowledge about the

effectiveness aloe gel in extending the shelf life of fruits, consumer resistance,

and economic impact of the aloe vera based edible coating. Second, there are

variety of edible coatings that are very popular used by traders to enhance the

quality of fresh fruits usage such as beeswax, carnauba wax and shellac.

Consequently, a more exhaustive study of the chemical, microbiological, and

physiological effects of the aloe gel based edible coating should be carried

out.

1.3 Significance of the Study

Nowadays, greater public awareness and international regulations have

emphasized the application of materials that show health benefits and are

environmentally friendly, especially in applications involving food products

(Arnon et al., 2015). One of the contributions of this study would be to show

the various benefits that people can get from aloe vera.

This study will provide a significant endeavor in using natural product coating

that is safe and environmental friendly as the alternative to synthetic films. As

stated by Siti Hazirah Fauzi & Noor Armylisas Abu Hassan (2017) synthetic

packaging materials are widely used for their ability in reducing mass, gas and

4
 soluble transfer between the foods and surrounding, but usually the synthetic

packaging are not biodegradable. In contrast with edible coating, it will give

some insight to society that aloe vera edible coating can contribute to

reduction of packaging waste, since they are considered as biodegradable.

Moreover, this study will be beneficial to human’s health. It is because Aloe

vera that applied for new coating poses no risk to human health and also they

are edible, odorless and does not affect the taste of fruits (Misir et al., 2014).

In addition, the use Aloe vera as the coating matrix may result in the increase

of nutritional value and health-promoting properties of fruit (Alberio et al.,

2015). The formulation of edible coating from Aloe Vera is hoped to be as

effective as synthetic films in maintaining the shelf life of the fruit. Besides,

the findings of this study will provide additional information or knowledge to

public regarding the benefits of using Aloe vera as the edible coating to

preserve the freshness of fruits.

1.4 Objectives of the Study

The objectives of the study are:

1. To determine combined effect of edible coatings incorporated with Aloe

vera on the shelf-life of fresh cut apples

5
2. To investigate the combined effect of edible coatings incorporated with

Aloe era on physiochemical and microbial properties of fresh cut apples.

3. To evaluate the sensory quality of fresh cut apples.

6
2.0 LITERATURE REVIEW

2.1 Aloe Vera (Aloe barbadensis miller)

Aloe vera is an important and traditional medicinal plant belonging to the

family Liliaceae (Rajeswari, et al., 2012). Aloe is a shrubby or arborescent,

perennial, xerophytic, succulent, pea-green color plant and the size of aloe

vera leaves is up to 20 inches long and 5 inches wide with triangular shaped

fleshy leaves that have spikes along the edges (Moghaddasi & Verma, 2011).

Each leaf consists of two basic products which is latex and an inner clear pulp.

For the latex, it is a bitter yellow exudate from the pericyclic tubules beneath

the epidermis of the leaf, it representing approximately 20-30% by weight of

the whole leaf and referred as “aloe juice” or “aloe sap” (Christaki & Florou-

Paneri, 2010). The inner leaf pulp makes up the majority of the plant by

volume, and is composed of large thin-walled parenchymal cells containing

Aloe vera gel (Guo & Mei, 2016).

2.1.1 Chemical composition of aloe vera

The chemical composition and the potency of the aloe vera are influenced by

many factors, for example like species or subspecies, climate, land and

irrigation, cultivation methods, harvesting, extraction processing, and storage

conditions (Guo & Mei, 2016). Aloe gel is 99% water and contains an

emollient polysaccharide, glucomannan which is a good moisturizer

(Christaki & Florou-Paneri, 2010). The gel also has pH of 4.5. Aloe vera latex

7
 is rich in anthraquinones, phenolic compounds, that have strong laxative

effects while they can also act as anti-bacterial especially against Gram-

positive bacteria, analgetics and antivirals (Christaki & Florou-Paneri, 2010).

Moreover, aloe vera contains 75 potentially active constituents which are

vitamins, enzymes, minerals, sugars, lignin, saponins, salicylic acids and

amino acids. These include lignin that have the ability of penetrating the

human skin, saponins which is an antiseptic property as well as foaming

agent, minerals, vitamins and amino acids (Raksha, Pooja, & Babu, 2014). In

some study, they performed the screening for phytochemical compounds in

aloe vera, the result show positive test in presence of tannin, saponin,

flavonoids and terpenoids but tests for steroids gave negative results

(Arunkumar & Muthuselvam, 2009).

2.1.2 Aloe vera in the context of coatings

In recent years, the use of Aloe vera gel has gained much attention for use as a

safe and environment-friendly postharvest treatments (Chrysargyris, Nikou, &

Tzortzakis, 2016). According to Chauhan, Gupta and Agrawal (2014), Aloe

vera can be used as edible coatings for fruits as its biological activities can

avoid loss of moisture, firmness, control respiration rate and maturation

development, reduce the time for oxidative browning, and minimize

microorganism proliferation. Moreover, the use of aloe as coating matrix may

result in the increase of the nutritional and health-promoting properties fresh

8
 fruits due to vitamins content inside the aloe gel. Recently, many research

have been done towards the efficacy of Aloe vera on maintaining several

fruits postharvest quality such as sweet cherry (Martínez-Romero et al.,

2006), grapes (Ali, Pandey, Singh, & Joshi, 2016), and peach (Hazrati, et al.

2017).

2.1.3 Antimicrobial efficacy of aloe vera

An antimicrobial is a substance that kills or inhibits the growth of microbes

such as bacteria, fungi, viruses or parasites (Misir, Brishti, & Hoque, 2014).

Fresh cut fruits are mostly susceptible to pathogenic and microorganism

exposure because of their surface is removed. Based on past study conducted

by Habeeb (2007), it shows that aloe vera efficient to inhibit the growth of

gram positive bacteria and gram negative bacteria. Another study also conduct

the potential of antimicrobial effect of aloe vera, it shows that aloe vera

extracts showed inhibitory action on the gram positive bacteria and Bacillus

subtilis except Bacillus cerus, and also inhibitory activity against gram

negative with the exception of Pseudomonas aeroginosa (Carac et al., 2016).

This is due to the composition of antimicrobial content in aloe vera. Radi et

al., (2017) listed the components in aloe vera such as saponins, acemannan,

and antraquinone derivatives are responsible for antibacterial activity.

9
2.2 Quality Attributes

Quality is a term which denotes a degree of excellence, a high standard or

value. The rising public demand for fresh-cut fruits and vegetable becomes

the increasing trend because of convenient and easy to eat food because

consumers can eat right out of the package after purchase the cut fruits and

vegetables (Barrett, Beaulieu, & Shewfelt, 2010). Due to that, quality criteria

for edible film coated fruits must be determined carefully, and the quality

parameters must be monitored throughout the storage period (Park, 1999).

Besides, fruit are known as the products that easily undergo enzymatic

browning, texture decay, microbial contamination and undesirable volatile

production if they are wounded (Tapia et al., 2008). So this perishable product

must be protected with good coating in order to maintain its freshness.

2.2.1 Appearance and color

In essence, the appearance of fruits and vegetable is the quality requirement

for every consumer and strongly affects the decision to buy. In addition,

another important factor in eye appeal of fruit is good color. For example,

bananas should be yellow in color without brown spots, tomatoes are red not

orange, cherries supposed to be red not yellow, and kiwifruit is green-fleshed

not yellow (Barrett et al., 2010). According to Barrett et al (2010), consumers

will evaluate visual appearance and color first, followed by the taste, aroma,

and texture. The major concern of the producer is the color that is the critical

10
 property of many fresh cut fruit usually for white flesh fruits such as apples

and pears (Toivonen & Brummell, 2008). The process operation of fresh cut

fruit causes damage and wounds to the fruit cell membrane thus it leads to

undesirable browning (Albanese, Cinquanta, & Di Matteo, 2007).

2.2.2 Texture

Consumers have clear expectations for the texture of fresh cut vegetables and

fruits. A research done by Harker, Gunson, & Jaeger (2003) shows that,

generally consumer preferences for apples are based on interactions between

texture and taste. For example, certain consumers most likely preferred a

sweet and hard apple, and some preferred a juicy and slightly acidic apple.

However, fresh fruits and vegetables with better maintenance in firmness,

crispness and crunchy texture are highly desired by consumers due to their

close association with tissue deterioration (Ma, Zhang, Bhandari, & Gao,

2017). Loss of firmness can be linked with tissue deterioration primarily due

to enzymatic degradation spaces, making the tissue firmer and apparently less

juicy (Yousuf, Qadri, & Srivastava, 2018). Thus, the most common way of

controlling softening phenomena in fresh-cut fruits is the use of treatments

with calcium salts.

11
2.3 Edible Coating

High perishability of minimally processed fruit and vegetables makes it

important to establish a method that can preserve these commodities for a

longer time period (Panwar & Mishra, 2015). The alternative of edible

coatings to provide active substances is one of the recent major advances

made in order to improve the shelf-life of fresh produce. Thus, edible coating

come out with an excellent method to carry additives since they are able to

maintain high concentrations of preservatives on the food surfaces, reducing

the impact of such chemicals on overall consumer acceptability of fresh fruit

(Oms-Oliu et al., 2010).

The use of edible coatings on fruits had been done since early 12th century

(Mahajan et al., 2018). Edible coating is any type of material used for

wrapping various foods to prolong shelf life of the product that may be

consumed together with food with or without further removal. Edible films

provide protection to substitute natural layers of fruit and vegetable to prevent

moisture losses, while selectively allowing for controlled exchange of

important gases, such as oxygen, carbon dioxide, and ethylene, which are

involved in respiration processes. A film or coating can also provide surface

sterility and prevent loss of other important components. Basically, its

thickness is not more than 0.3 mm (Embuscado & Huber, 2009).

12
 Mahajan et al. (2018) states, from an environmental perspective, replacement

of plastic films with edible or biodegradable materials are desirable

considering the concerns about the use of plastic materials as coatings. So, it

is important to replace non-food-grade materials by bio-based and

biodegradable food-grade materials in the particular case of the food sector.

Coatings can be formulated from plants extracts such as polysaccharides,

proteins and lipids, Aloe vera, essential oil alone or in combination. These can

either be applied as thin film to form wraps or pouches, or as coatings on food

(Mahajan et al., 2018). From this, edible coating can establish as an

alternative source to synthetic packaging due its biodegradability, preventing

environment-related problems.

2.3.1 Gelatin films

Gelatin is formed when collagen is exposed to a medium heat treatment under

acidic or alkaline conditions (Embuscado et al., 2009). Gelatin is obtained by

controlled hydrolysis from the fibrous insoluble protein, collagen, which is

widely found in nature as the major constituent of skin, bones and connective

tissue (Bourtoom, 2008). Gelatin has been used as a coating by the food and

pharmaceutical industry for years. Gelatin coatings show good barrier

characteristics against oxygen and aroma transfers at low and intermediate

relative humidity. However, they have poor barrier properties against water

vapor transfer due to their hydrophilic nature (Andrade et al., 2014). In

13
 addition, gelatin is used to encapsulate low moisture or oil phase food

ingredient and pharmaceutical. Such encapsulation provides protection against

oxygen and light, as well as defining ingredient amount or drug dosage

(Bourtoom, 2008). According to Ramos, Valdés, Beltrán, & Garrigós (2016),

different types of additives could be added to improve or modify the final

properties in order to achieve suitable gelatin-based films or coatings for food

packaging.

2.3.2 Calcium chloride

Maintaining the fruit texture in order to prolong the shelf-life and maintaining

good flavor is very important because fruit firmness usually influences the

consumer perception about the freshness of the fruits and its storage

characteristics. Calcium level in the fruit tissue is related to the softening and

firmness of fresh-cut fruits (Silveira, Aguayo, Chisari, & Artés, 2011). For

this reason, calcium dips have been employed to improve firmness and extend

the postharvest shelf-life of a wide range of fruit and vegetables (Hernández-

Muñoz, Almenar, Ocio, & Gavara, 2006). According to Liu et al., (2009) the

firmness of fruit increase after calcium chloride treatment. This is due to

increase of the binding of the cell wall polysaccharides by calcium chloride.

Calcium ions form cross-links or bridges between free carboxyl groups of the

pectin chains, resulting in strengthening of the cell wall (Martı´n-Diana et al.,

2007). In addition dipping and vacuum or pressure infiltrations are common

14
 techniques used to increase cell wall calcium content of fruit tissue after

harvest (Hernández-Muñoz et al., 2006). Chong, Lai, & Yang (2015) also

proved that the combined effect of calcium chloride with the addition of

chitosan provide the highest firmness of honey dew melon.

2.4 Acidifying Agent

Acidifying agent plays an important role to prevent enzymatic browning.

Browning effect on the fresh fruit is a main factor to reduction in quality and

acceptability of fresh fruits by consumers. The phenomenon is usually caused

by the enzyme polyphenol oxidase (PPO), which in presence of oxygen,

converts phenolic compounds into dark colored pigments (María Alejandra

Rojas-Graü, Soliva-Fortuny, & Martín-Belloso, 2009). The substrates

involved in these reactions are located in the vacuoles while enzymes are in

the cytoplasm and the reactions can take place only if they are mixed and in

the presence of oxygen (Ioannou & Ghoul, 2013). Basically, browning

reaction that occurs in food is divided into two types which are enzymatic and

non-enzymatic browning. Most browning reactions occurred in fresh fruits

and vegetables are associated with the enzymatic browning type (Supapvanich

et. al., 2016). However, Suttirak & Manurakchinakorn (2010) states that, the

role of the acidifying agent is to inactivate polyphenol oxidase (PPO) by

lowering the pH of the product to minimize the activity of PPO. They also

propose that PPO has little activity at pH values below 4 due to the loss of

15
 cooper at the active site (Suttirak & Manurakchinakorn, 2010). The main

acidifying agents that had been used in edible coating are ascorbic acid and

citric acid.

2.4.1 Ascorbic acid

Ascorbic acid acts as reducing agent in fruit. According to Rojas-Graü et al.,

(2009) ascorbic acid are responsible for the reduction of the o-quinones,

generated by the action of the PPO enzymes, back to the phenolic substrates.

In addition, ascorbic acid (L-ascorbic acid) and its various neutral salts and

other derivatives have been the leading generally recognized as safe (GRAS)

antioxidants for use on fruit and vegetables and in fruit juices (Rico, Martín-

Diana, Barat, & Barry-Ryan, 2007). Besides, the effect of the ascorbic acid

have been the most studied for the use in processed foods (Cortez-Vega,

Becerra-Prado, Soares, & Fonseca, 2008). Several thiol containing

compounds such as cysteine, N-acetylcysteine, and reduced glutathione also

can be used an alternative to ascorbic acid in enzymatic browning studies

(Rojas-Graü, Soliva-Fortuny, & Martín-Belloso, 2008).

16
2.4.2 Citric acid

Citric Acid acts as the chelating agent in the edible coating (Suttirak &

Manurakchinakorn, 2010). According to Rojas-Graü et al., (2008) carboxylic

acids such as citric acid have been also suggested as effective antioxidant

agents in fresh-cut fruits. Citric acid is generally recognized as safe chemical

preservatives and compared to other organic acids, citric acid is relatively

cheap and easily obtained (Chiumarelli et.al, 2010). Collaboration of citric

acid with ascorbic acid shows potential to inhibits the enzymatic browning

and maintain the quality of fresh cut apple in research done by Jo et al.,

(2014).

2.5 Synthetic Coating on Fruits

Evidence in the recent years indicates that the use of morpholine in synthetic

fruit coatings in the United States is common and the usage of ammonia can

be considered as alternative instead (Hagernmaier, 2004). However, consumer

trends nowadays prefer more natural product because the utilization of

synthetic coating has unfavorable effect on environment and health as

morpholine that are water soluble can lead to formation of carcinogenic

compound N-nitrosomorpholine (NMOR) which can enter our body trough

absorption through the skin and ingestion if not properly handled or consumed

directly (Kumar & Kapur 2016). Hagernmaier (2004) added that ammonia-

17
based coatings are difficult to prepare because ammonia is highly volatile and

its vapors are unpleasant and toxic.

In addition, a section of irresponsible traders used calcium carbide in fruits to

ensure the fruits are fresh and ripen faster (Misir et al., 2014). Calcium

carbide is a chemical compound that will reacts with water to produce

acetylene in ripening process (Dhembare, 2014). The use of carbide gas from

calcium carbide is can cause bad effect to the human body as calcium carbide

also contains traces of arsenic and phosphorus (Rahim, 2014). Rahim, 2014

added that fruits ripened with calcium carbide are soft with good skin color

but the quality of flavor is poor and also they have a shorter shelf-life.

18
3.0 METHODOLOGY

3.1 Materials

3.1.1 Raw materials

Aloe vera leaves will be obtained from Kuala Lipis, Pahang and will be

harvested from the mature plant. Red apples (Red Delicious) are purchased

from a local wholesale market in Jengka, Pahang and will be stored in a

refrigerator at 4°C for one day until processing. Size uniformity and peel

coloration will be carefully selected before purchase to eliminate the damaged

ones.

3.1.2 Chemicals

Gelatin, calcium chloride, citric acid, ascorbic acid

3.1.3 Apparatus

Blender, 2000 rpm centrifuge, Agar plate, Refrigerator, pH paper, Magnetic

stirrer, Precision balance, conical flask, beaker

19
3.2 Methods

3.2.1 Sample Preparation

Apple with uniform size and shape and without any mechanical damage will

be selected. Apples will be sanitized by immersion in chlorine solution (300

ppm). Then the apple will be rinsed with distilled water and dried at room

temperature. All utensils and surfaces used during cutting also will be

sanitized with 300 ppm chlorine solution. Cut the apple into slices with a

knife with uniform size.

3.2.2 Preparation of aloe vera gel

Aloe vera extract will be obtained from fresh aloe vera leaves with absence of

visual defects. Matured leaves of Aloe vera will be washed with a mild 25%

chlorine solution. The upper epidermis of aloe vera will be removed and

inner mucilaginous material will be cut into small cubes then blender the gel

(Panwar & Mishra, 2015). The the mixture will filtered to remove fibers. The

gel matrix will be pasteurized at 70oC for 45 min. The gel will cool

immediately to an ambient temperature for 5 minutes. 2.0 g/L ascorbic acid

will be added then, 4.5 g/L citric acid will be added to maintain the pH at 4

(Adetunji et al., 2012).

20
3.2.3 Preparation of calcium chloride solution

0.5% w/w calcium chloride at will be dissolved in sterile distilled water at

room temperature.

3.2.4 Preparation of gelatin solution

Gelatin solution will be prepared by adding 1% w/w gelatin at in sterile

distilled water previously heated at 50°C on a stirring hot until gelatin was

completely dissolved.

3.2.5 Coating the apple slices

Before coating, all the formulations will be cooled to ambient temperature

(Sharmin et al., 2015). A five-step procedure (layer-by-layer) will be used to

ensure the proper coating of the fruit pieces. At room temperature

approximately 27°C, apple pieces will be dipped into each coating solution for

1 minute. Then, the excess coating solution will be allowed to drip off for 1

minute, just before submerging the apple pieces for additional 1 min in the

solution of calcium chloride.

The order of the coating solutions was as follows: calcium chloride, aloe vera

with ascorbic acid and ascorbic acid, calcium chloride, gelatin, and calcium

chloride. Next, the coated apple slice will be drained for 30 min. This step

will be repeated for different concentration of aloe vera 50%, 100% and

150%. Control samples (uncoated) only dipped into sterile distilled water for

1minutes and then allowed to drip off 2 minutes before further analysis. The

21
 prepared apple slices will be placed in polyethylene terephthalate (PET)

clamshells and will be store at 4°C within 14 days.

3.3 Product Quality Attribute Analysis

3.3.1 Measurement of Titratable Acidity (TA)

The test will be performed in triplicate at room temperature. Three pieces per

treatment (coated samples and controls) will be used. The apple slices will be

homogenized in a blender and the blended puree will be used to determine

titratable acidity (TA). The homogenate will be centrifuged at 2000 rpm for 1

min to obtain a clear juice.

TA values will be determined by diluting each 10 mL aliquot of homogenized

apple juice in 50 mL of distilled water and titrated by using 0.1 M NaOH.

Results will be expressed as mg of ascorbic acid per gram of sample.

3.3.2 Weight Loss Determination

Weight loss will be evaluated by weighting all samples with a precision

balance at the initial of storage (day 0) and at day 2, 6, 9 and 13. The

difference between the two values will be considered as the weight loss. The

percentage of weight loss was determined by the following equation: (Song et

al., 2013)

22
Weight Loss Percentage =

(Initial weight − weight in the specific time of storage) X 100

Initial weight

3.3.3 Sensory Evaluation

Sensory evaluation will be performed immediately after the apple slices are

removed. 5 students will be chosen to evaluate the samples by visual

inspection of color, odor, texture, firmness and overall quality for days 0, 2, 6,

9 and 13 of storage at 4 ºC.. The students will scored the samples using a five-

point hedonic scale, where a score of 1 represents attributes most dislike and a

score of 5 represents attributes most like. Scores higher or equal to 5 will

consider as acceptable.

Sensory
Sensory Scores
Characteristic
1 2 3 4 5
Dislike Dislike Neither Like Like
Color
extremely moderately nor moderately extremely
dislike
Dislike Dislike Neither Like Like
Odor extremely moderately nor moderately extremely
dislike
Dislike Dislike Neither Like Like
Texture extremely moderately nor moderately extremely
dislike
Dislike Dislike Neither Like Like
Firmness extremely moderately nor moderately extremely
dislike
Dislike Dislike Neither Like Like
Overall
extremely moderately nor moderately extremely
acceptability
dislike

23
 Figure 3.5: Five-Point Hedonic Scale Rubric (Lim, 2011)

3.3.4 Microbial Evaluation

Standard plate count will be used to estimate the total aerobic count of apple

over storage time. The apple samples will be evaluated for bacterial count on

days 0, 2, 6, 9 and 13. Two apple pieces in each treatment will be smashed

inside a sterile plastic bag with 90 ml of 0.85% NaCl. 1 mL of juice sample

will be serially diluted to 10-2 dilution in saline (0.85% NaCl w/v) and wil be

spread on agar plate for analysis (Nayik, Amin, & Bhat, 2013). Petri dishes

will be incubated at 37°C for 48 hours. The mean of triplicate plates are a

single measurement per package; bacterial growth will be calculated as

follows:

BG = Log Nf
No

Where Nf, is the colony forming units per gram at time t of shelf life; and No

is the colony forming units per gram on day 0 of shelf life.

3.4 Statistical Analysis

Data analysis will be performing using SPSS software. Apple properties as

well as differences among experiments will be evaluated. Differences between

variables were tested for significance by one-way ANOVA using Duncan's

multiple range tests. Statistical significance was expressed at the P<0.05 level.

24
CITED REFERENCES

Albanese, D., Cinquanta, L., & Di Matteo, M. (2007). Effects of an innovative

dipping treatment on the cold storage of minimally processed Annurca

apples. Food Chemistry, 105(3), 1054-1060.

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