ABSTRACT

COMPARATIVE EVALUATION OF THE PHYSICOCHEMICAL AND

SENSORIAL QUALITY OF DRAGON FRUIT JAMS MADE WITH

DIFFERENT TYPES OF CITRUS JUICES AS ACIDULANTS

CHAI CHENG KAI

The processing of dragon fruit into jams has emerged as a promising solution

against the short shelf life of the fruit at ambient temperature that could limit its

commercial success. As the quality of dragon fruit jam is heavily influenced by

the acidulant used and with the growing preference for natural food additives

among the consumers, the potential use of citrus juices as acidulants in jam

making was explored in this study. With the aim of comparing and evaluating

the physicochemical, sensorial and microbiological quality of dragon fruit jams

made with orange, lemon and calamansi juices, the three jam treatments were

analysed for their physicochemical characteristics, which included colour,

moisture content, pH, firmness, total soluble solids content and viscosity, via a

variety of instrumental methods, and their likability in terms of aroma,

appearance, taste, mouthfeel and overall acceptability was assessed through

sensory evaluation. The jam treatments were also subjected to microbiological

analysis where their total plate count was determined for a period of three weeks.

The results revealed that orange, lemon and calamansi juices gave rise to dragon

fruit jams with rather similar physicochemical properties, except for their pH,
where orange juice gave rise to jams with a significantly higher pH (4.55)

compared to the other treatments, and viscosity, where each jam treatment was

found to have a viscosity that was significantly different from each other. The

jam treatments were also found to have similar sensorial qualities, where the

major differences were the significantly lower acceptability of dragon fruit jams

made with orange juice in terms of their taste (5.81) and overall acceptability

(6.07), as compared to those of the jams made with calamansi juice, which were

6.64 and 6.83, respectively. Additionally, it was demonstrated that all jam

treatments were able to remain microbiologically stable for three weeks.

 CHAPTER 1

INTRODUCTION

1.1 Background Study

Ever since the dawn of mankind, fruits have undoubtedly become an integral

component of man’s sustenance, with efforts being continuously made in order

to have these sources of nourishment well preserved long after their harvesting.

Amid the vast variety of preserved fruits that our ancestors had successfully

fabricated, jams, which might have arisen from a primeval attempt on fruit

preservation, comprise products of undeniable significance (Vibhakara and

Bawa, 2006). Being a technique of fruit preservation that precedes canning and

freezing, as according to Peckham (1964 cited in Vibhakara and Bawa, 2006,

p.189) and Thakur, et al. (1997 cited in Vibhakara and Bawa, 2006, p.189), jam

preparation, as a method of fruit preservation that was employed during the

harvesting season of fruits, was initially established as an artistry by the

homemakers. These products of fruits would then become more popular and

obtainable following the increment in affordability of sugar production

(Vibhakara and Bawa, 2006).

Today, a number of specifications and regulatory standards, to which jam

products manufactured at large scale must show compliance so as to avoid being

deemed to be misbranded, have been established. To the jam manufacturers, the

key to producing jams that are consistently conforming to the quality

requirements has always been the right utilization of understanding towards

pectin, as well as the principles underlying the generation of the gel matrix of

pectin, sugar and acid. Whilst preparation of jam products involves the

employment of dissolved solids of an enormous concentration, pectin and sugar

alone would not be enough to give rise to the final products (Vibhakara and

Bawa, 2006). Instead, the acidity of the fruit possesses an equal importance as

well, as according to Breverman (1963 cited in Vibhakara and Bawa, 2006,

p.189), and this aspect of jam making is well managed by what is known as an

acidulant.

Acidulants have been utterly essential in bringing the pH of the jam into the

range that is necessary for the formation of gel, and optimizing the sensory

properties of the final product (Bourne, 2017). Being either acids that are

inherent in fruits or those that are added to the fruits later in the processing,

acidulants have been versatile regardless of their form, such that these

compounds have had critical roles in preservation, flavouring and so on, other

than their main function of acidification (Vibhakara and Bawa, 2006).

Owing to the fact that no two acids would possess identical physicochemical

properties, selection of a suitable acid as an acidulant in jam making is cardinal.

Whereas the behaviour of each acid is not dependent upon pH and there is no

necessary preference of one particular acid over another, citric acid is the

predominant acidulant employed in the production of most fruit jams

(Vibhakawa and Bawa, 2006). This leads to the potential application of citrus

juices as acidulants in jam making, as the organic acid content in the juices of

citrus fruits has been found to be comprising of predominantly citric acid, along

with malic and succinic acid (Vandercook, 1977 cited in Liu, Heying and

Tanumihardjo, 2012, p.534). On top of that, the already existing application of

citrus juices in food industry as additives, as suggested by He, et al. (2011 cited

in Lv, et al., 2015, p.1) and Kelebek and Selli (2011 cited in Lv, et al., 2015,

p.1), has further boosted such potential. In this study, juices of orange, lemon

and calamansi had been chosen as three separate sources of acidulants for jam

making as oranges and lemons are the most widely known instances of citrus

fruits with considerable significance in commerce, whereas calamansi are citrus

fruits of economic importance in Malaysia as well as other Southeast Asian

nations (Liu, Heying and Tanumihardjo, 2012; Hoyle and Santos, 2010).

In this study, the effect of different types of citrus juices as acidulants on the

attributes of the final jam product would be investigated via the making and

analysis of dragon fruit jam. Dragon fruit had been selected as the type of fruit

from which jams would be derived as the quality and features of dragon fruit

has made the fruit crop highly demanded in the domestic market of countries

where the fruits are produced, and the international market (Le Bellec, Vaillant

and Imbert, 2006; Ortiz-Hernández and Carrillo-Salazar, 2012). In spite of its

status in the markets around the globe, dragon fruit could have its commercial

success restricted by its short shelf life, as it is only able to last for 3 to 4 days

at room temperature (Le Bellec, Vaillant and Imbert, 2006). This undesirable

attribute of dragon fruit can be easily overcome by having the fruit processed
into jams, whose quality, in turn, is significantly influenced by the acidulant

added therein. Besides, as the flesh of dragon fruits has been found to be of low

acidity, as according to Le Bellec, Vaillant and Imbert (2006), the acidulant

would be a crucial factor that determines whether jam can be successfully

produced from this type of fruits.

The aim of this study was to compare and evaluate the effect of different types

of citrus juices, which are orange, lemon and calamansi juices, as acidulants on

the physicochemical, sensorial and microbiological qualities of dragon fruit jam.

1.2 Objectives

The objectives of this study were:

i. To compare the physicochemical properties of dragon fruit jam

made with different types of citrus juices (orange, lemon and

calamansi juices) as acidulants.

ii. To evaluate the sensory properties of dragon fruit jam made with

different types of citrus juices (orange, lemon and calamansi

juices) as acidulants.

iii. To determine the microbiological quality of dragon fruit jam

made with different types of citrus juices (orange, lemon and

calamansi juices) as acidulants.

 CHAPTER 2

LITERATURE REVIEW

2.1 Jam

2.1.1 Legal Definition, Essential Composition and General Requirements

for Jams

Jam, as defined by Codex Alimentarius Commission (2017 p.2), is the product

that is derived from at least one type of fruits by mixing the fruit, which can be

in the form of whole fruit, sliced fruit, fruit pulp or puree that have or have not

been concentrated, with sweeteners and subsequently, having the consistency

of the mixture adjusted appropriately with or without water addition. According

to this definition, jams must have been composed of two ingredients, namely

the fruit ingredient and the sweetener, and these ingredients have also been

strictly defined by Codex Alimentarius Commission (2017 p.2) as well. For the

fruit ingredient, Codex Alimentarius Commission (2017 p.3) has specified that

the fruit content of finished jam product shall be at least 45% or 35% in general,

with exception given to certain fruits. In the case of mixed fruit jam, reduction

of the minimum fruit content must be done in accordance with the percentages

of the fruits used. Sweeteners, on the other hand, have been defined as (i) all

sugars that are defined by Codex Alimentarius Commission in its standard for

sugars, (ii) fruit sugars, (iii) syrup of fructose, (iv) brown sugar and (v) honey,
which Codex Alimentarius Commission has defined in its standard for honey.

Codex Alimentarius Commission (2017 p.4) has also permitted the use of any

other proper ingredients of plant origin, such as herbs and spices, provided that

these ingredients are not added with the intention of disguising poor product

quality and deceiving the consumer.

Codex Alimentarius Commission (2017 p.4) has mandated that the finished jam

products shall have soluble solids content of 60 to 65%. The upper limit of

permitted soluble solids content for jams has not been restricted to 65%, as it

can be greater depending on the federal legislation established in the nation

where the finished products are retailed. The end product shall possess a proper

gel consistency in addition to a colour and flavour that are not deemed to be odd

to the type of fruit to which the jam is derived, while having the flavour and/or

colour imparted by any other ingredients added taken into account. Also, it shall

not contain any defective materials that are commonly found in fruits (Codex

Alimentarius Commission, 2017 p.4).

2.1.2 Common Constituents of Jam

A jam is typically comprised of four fundamental components, and since it

originated as a means of fruit preservation, the jam would undoubtedly have

fruits as its main ingredient. According to Vibhakara and Bawa (2006), jams

can be derived from five distinctive forms of fruits, which are (i) fresh fruits, (ii)

fruits under cold storage, (iii) fruits that have received thermal treatment, (iv)

fruits that are treated with sulphur dioxide, and (v) dried or dehydrated fruits.
Whilst all these forms of fruits can be processed into jams, it is claimed that

jams of the highest quality are generally given rise by fresh fruits (Vibhakara

and Bawa, 2006). Regardless of the claim, it is always recommended to choose

high quality fruit to work with, no matter which form of fruit is intended to be

processed into jam, as the quality of fruit would be the primary determinant of

that of the finished product. The selection of fruit of superior quality is also

essential because it is mostly the original fruit that contributes to the colour and

flavour of the jam (Featherstone, 2016).

Another major ingredient of jams, apart from fruits, would be sweetening agents.

Sweetening agents have been employed in the manufacturing of jams for a

number of reasons other than supplementation of sweetness in the finished

product (Vibhakara and Bawa, 2006). Other than giving jams their body and

bulk, sweeteners are held accountable for the texture of jams as these substances

control viscosity of jams by aiding in the formation of gel within the fruit

preserves. On top of that, the hygroscopic characteristic of the sweetening

agents has also made them an eligible preserving agent as they could bind to

moisture in the jam that is needed by the microbes for survival and growth

(Vibhakara and Bawa, 2006).

Being one of the important factors that govern the formation of the gel, gelling

agents are also an essential component in the making of jam. According to

Fishman and Jen (1986 cited in Vibhakara and Bawa, 2006, p.191), a gelling

agent would be considered ideal if the addition of this gelling agent in a product
does not lead to the disruption of product aroma or flavour. Whereas multiple

types of gelling agent, such as gelatin and agar, are applicable in the production

of jams, pectin remains to be the most common one used in fruit and jam

processing (Vibhakara and Bawa, 2006). Addition of commercial pectin is

normally necessary to ensure the uniformity of jam consistency, as the amount

of pectin that occurs naturally in fruits is varying depending on the type of fruit,

its degree of ripeness during the period of processing, and the environmental

conditions (Featherstone, 2016). Under suitable conditions, pectin would form

gels when it has been dissolved in water. Being a colloid with negative charges

in a fruit substrate that is acidic, pectin would establish a fibrous network with

the capability of holding up liquids, following the disruption of pectin-water

equilibrium as resulted from the addition of sugar to the colloid. This fibrous

network would then give rise to the gel that is critical for jams (Smith, 2003).

The fourth and the final fundamental constituent of jams would be the acidulants.

Acidulants are added in jams for various purposes. Other than the main function

of acidification, acidulants are also essential in pH regulation and maintenance,

preservation, flavour enhancement, metal chelation, gel formation and

inhibition of oxidation in jams (Vibhakara and Bawa, 2006). These functions of

acidulants in jams will be discussed in detail in the later section.

2.1.3 Basic Steps of Jam Making

Though it may have been viewed as a process with definite simplicity, the

processing of jam would nonetheless be requiring extensive control and strict

adherence to the scientific approaches in each and every step of manufacture.

Whilst a number of methods have been applicable for the making of jam, all of

these methods would have processed fruits into jam through the basic steps of

fruit preparation, boiling, and finishing (Vibhakara and Bawa, 2006). The

process of jam manufacturing begins with the selection of suitable fruits for

processing, where fruits with full maturity, rich flavour and desirable texture are

desirable. The selected fruits would have their surface cleaned with water to

remove any of the dirt present. In the case where the residues of lead or arsenic

spray may be present on the fruit, the fruit should then be sanitized in a warm

1% hydrochloric acid solution before being rinsed in water (Giridhari Lai, et al.,

1986 cited in Vibhakara and Bawa, 2006, p.198).

Fruit selection would be crucial to the finished jam product as it determines the

fruit-to-sugar ratio that is suitable for employment in jam manufacturing

(Vibhakara and Bawa, 2006). This ratio is dependent upon the fruit variety, the

degree of ripeness of the fruit, and the desired effect, though the ratio of 1:1 is

commonly employed in most fruit jams as it is suitable for a wide range of fruits

including berries and pineapples. Conversely, for fruits that are sweet and have

low acidity, lesser amount of sugar would have been required (Vibhakara and

Bawa, 2006).

The next step, boiling, is one of the most vital steps in the making of jam. As

boiling is performed with the chief objective of concentrating sugar to enable

gel formation by means of evaporating the excess moisture present in the

mixture, fruits that display resistance towards boiling are desired (Ashish

Kumar, 1988 cited in Vibhakara and Bawa, 2006, p.199). During the process of

boiling, skimming of the fruit pulp should be done when necessary for the sake

of removing coagulated material and thorough mixing should be ensured by

constant stirring of the mixture. Boiling will be continued until the jam has

obtained a desirable consistency, and it is crucial to exert adequate control over

this process so as to avoid prolonged boiling that will lead to excessively

concentrated soluble solids, excessively inversed sugars and hydrolysed pectin

(Vibhakara and Bawa, 2006).

Subsequently, the boiled jam would be subjected to finishing which is

comprised of three major steps, namely pre-cooling, filling and cooling of filled

jam. According to Rauch (1965 cited in Vibhakara and Bawa, 2006, p.200),

cooling of the jam prior to filling should be done until the jam is close to its

setting point, and extensive control is essential to prevent the limit from being

exceeded so that disruption of the gel and curdling of the jam would not occur.

After that, the cooled jam would then be filled into the glass jars. Filling should

be done in a manner that results in a headspace of 1.25 cm at most for each filled

glass jar. Loose placement of the jar lids on the jars should be done as soon as

filling has taken place, and these lids should be tightened within 2 to 3 min after

the filling process. This would allow the exhaustion of the air from the

headspace of each filled jar. Following the filling process, the filled jam must

not be cooled in a hasty manner. Cooling of the glass jars and their content has

to be done by air cooling and it is carried out until the jam product has been well

set. Upon cooling, condensation of the steam in the headspace of the filled jar
will also take place, and this will result in the creation of a hermetic seal that

can largely prevent the deterioration of the jam product during storage

(Vibhakara and Bawa, 2006). This concludes the basic steps of jam

manufacturing.

2.2 Acidulants

2.2.1 Functions of Acidulants in Jams

2.2.1.1 Acidification and pH Regulation

Acidulants are crucial in the processing of jam where the pH of the jam affects

the formation of the gel and determines whether the gel formed would be of

optimum characteristics and strength (Deshpande, Deshpande and Salunkhe,

1994). Use of acidulants for the regulation of pH of the jam is especially

essential to ensure the success in gel formation whenever pectin, particularly

pectin with high degree of methoxylation (HM pectin), is used as the gelling

agent for jam making (Baker, et al., 2004). This is because whilst the properties

of gel are influenced by the concentrations of sugar, pectin and acid, mass

production of jam will usually have the batches of jam cooked to an established

total soluble solids content and with a fixed amount of pectin, leaving the

acidulants to be the sole determinant of the gel character. Through pH reduction,

these acidulants would drive the formation of a network of water and solute

trapping lattice from the precipitated molecules of pectin. They propel the
formation of such a network by reducing the electrostatic repulsion between the

adjacent, negatively charged pectin chains and facilitating of hydrogen bonds

formation among them (Featherstone, 2016). This network of lattice would then

give rise to the gel that gives the jam its body. In addition, via pH reduction,

acidulants would also allow the achievement of sterilization and inactivation of

spores in jam products at a lower temperature and/or a shorter of period of time,

thus minimizing the loss of flavour and firmness owing to degradation of

components and structures in jam upon thermal treatment (Dziezak, 2003).

2.2.1.2 Preservation and Microbial Inhibition

Acidulants have also been significant in the preservation of jam as they are

capable of preventing or retarding microbial growth as well as bacterial spore

germination that will result in the spoilage of jam or even cause foodborne

disease in consumers (Dziezak, 2003). According to Dziezak (2003), whilst

both the pH-lowering effect of the acidulants as well as the action of the acids

in their undissociated state are accredited for this effect, it is widely known that

the latter plays a predominant role in exhibiting the antimicrobial activity of the

acidulants. The undissociated acids carry out such function by penetrating

across the cell membrane of microbes and inhibiting microbial growth by

dissociating within the interior of the microorganisms and thus, interfering with

the pH gradient that was maintained across the plasma membrane of the

microbes (Raju and Bawa, 2006). It is also worth mentioning that the pH-

lowering effect of the acidulants would enhance the activity of the undissociated

acids by shifting the equilibrium in a manner that favours the latter. Also, in the
case where preservatives like benzoates are added to jam, acidulants would

enhance the antimicrobial activity of these preservatives by maintaining the pH

of jam at a range that is optimum for their actions (Dziezak, 2003).

2.2.1.3 Flavour Enhancement

Acidulants are also incredibly important for the modification of the flavour of

jam, where their absence would have rendered the product to have an utterly flat

taste. Acidulants allow the development of the flavour of the jam up to its full

potential by imparting sourness or tartness that is necessary to have the

immoderate sweetness brought by the sweetening agent balanced off properly

(Deshpande, Deshpande and Salunkhe, 1994). With acidulants, the sweetness

to sourness ratio, which is also known as the ratio of brix to acid, can be adjusted

even though it is close to balance in order to have the chief flavour of the jam

expressed and those peripheral flavour notes enhanced so that they would not

be concealed by surplus sourness or sweetness (Sausville, 1974 cited in

Deshpande, Deshpande and Salunkhe, 1994, p.14). In addition, acidulants are

also capable of modifying or intensifying the sensations of taste brought by

flavour components that are distinct from themselves, mixing taste

characteristics that are not related to each other, and masking the unpleasant

aftertaste by having the sourness sensation that they impart to be prolonged

(Dziezak, 2003).
2.2.1.4 Metal Chelation

The application of acidulants in jam making also serves a purpose in the

prevention of oxidative reactions that take place naturally in jam products and

give rise to unwelcomed results including unpleasant changes in colour as well

as loss of nutrients and flavour (Dziezak, 2003). The ability of acidulants to

inhibit the occurrence of oxidation in jams is attributed to their metal chelating

activity, where these acidulants would bind or confine metallic ions that are

present in jams and are the catalysts for these oxidation reactions (Deshpande,

Deshpande and Salunkhe, 1994). Chelation of these metallic ions by the

acidulants would have rendered them to be unavailable for the oxidation

reactions, and with that, these reactions would be inhibited successfully.

Moreover, when acidulants are employed in conjunction with antioxidants, they

would also exert a synergistic effect on the activity of these antioxidants and

thus, further improving the stability of jam products (Dziezak, 2003).

2.2.1.5 Nutrient Fortification

Enhancement of the nutritional value of jam products can also be done with

acidulants that have incorporated ascorbic acid in them. Conventionally and

preferentially, oxidation of the ascorbic acid that is present in jams to

dehydroascorbic acid would take place, and its occurrence would be associated

with the swift depletion of the activity of vitamins in jams. This reaction can

only be reversed if ascorbic acid occurs in a substantial quantity in jams. With

the employment of acidulants, the preservation of ascorbic acid in jam products

is enabled as the acidulants are capable of driving the reversal reaction by

imparting a great degree of acidity in jams that is necessary for the maintenance

of a low degree of redox potential that does not favour the oxidation of ascorbic

acid. With that, the recovery of ascorbic acid from its oxidized form is made

possible, and the nutritional value of jam products is preserved (Desphande,

Deshpande and Salunkhe, 1994).

2.2.2 Influence of Acidulants on the Overall Quality of Jams

The efficacy of various acidulants in achieving their function of optimising

various quality attributes of jams had been indirectly evaluated in a number of

studies. Ferreira, et al. (2004), who examined a total of eighteen different

branded quince jam products that were commercialized in the markets of

Portugal in relation of their physicochemical, sensorial and microbiological

qualities, had found that the acidulants would not be as influential as the nature

of the raw fruit materials and the parameters established for the jam production

were, on the sensorial properties of the jam products assessed. This conclusion

had been made from their discovery that the sensorial quality of the different

quince jam products was not correlated with the organic acid profile of these

products at all. Ferreira, et al. (2004) went on to infer that it was the manner in

which the quince pulp was processed that would be the sole determinant of the

quality of the final quince jam product. Whilst the findings and inferences of

Ferreira, et al. (2004) are indeed solid, it is noteworthy that the study was not

able to determine if the organic acids that were detected from the samples were

naturally occurring compounds in the quince pulp or were the acidulants added

during the processing of quince jam, thus making the statement made by
Ferreira, et al. (2004) against the effect of acidulants on the quality of jam to be

inconclusive.

The inference made by Ferreira, et al. (2004) against the efficacy of acidulants

in jam quality optimisation was later overthrown by the findings of Khan, et al.

(2017) in their research on the quality of the newly developed jam that they

produced from seven different formulations, three of which incorporated 0.2 g

of citric acid as the acidulant, throughout three months of storage at ambient

temperature. The results of the experiment conducted by Khan, et al. (2017)

showed that jams made from two of the three formulations that employed citric

acid as the acidulant retained their desirable quality best throughout the storage

period. The results had also shown that although jams made with the acidulant

were not significantly better than those that were made without the acidulant in

terms of their quality characteristics at the beginning of the storage period, the

acidulant would later be proven to be essential for the maintenance of the jam

quality as it minimized the undesirable quality changes that took place in the

jams along their storage (Khan, et al., 2017). With these findings of Khan, et al.

(2017), the importance of acidulants on the quality of jams have been effectively

verified.
2.3 Citrus Juices

2.3.1 Demonstration of the Capability of Citrus Juices as Acidulants in Jam

Making

2.3.1.1 Existing Application of Citrus Juices in Food Production

The versatility of a number of citrus juices has been demonstrated by the

employment of these citrus juices in the production and preparation of various

types of food. Among the citrus juices that are utilized in the food and beverage

industry, lemon juice might have been the most prominent one. Whilst the high

acidity of lemon juice has made it less popular for consumption in its pure form,

it is the same property that grants the juice its miscellaneous uses in the

industries. According to Goodrich (2003), lemon juice has been being a

common component in a large number of foods and beverages that are

processed and produced on a commercial scale, most famously as a constituent

in juices. Apart from that, its tartness, tanginess and freshness has also made it

a valuable component in the recipes of a number of home-made foods (Goodrich,

2003).

Similar to lemon juice, calamansi juice has also been useful in the preparation

and production of a variety of foods owing to its acidity. In Asian nations, the

great acidity of calamansi juice has rendered it to be a popular component in the

production of numerous beverages, including mixes of juices, sweetened

beverages that are flavoured with fruits, as well as carbonated drinks

(Ramadugu, et al., 2017; Hoyle and Santos, 2010). On top of that, it is also a

regular constituent in marinades and salad dressings (Bhat and Paliyath, 2016).

In certain nations, people have also been using calamansi juice as an additive

for the preparation of various foods (Abdullah, Ch’ng and Yunus, 2012).

As for orange juice, despite this particular citrus juice has yet to have

noteworthy application in the food and beverage industry, its potential for

utilization as an acidulant in jam making remains promising considering that

the acids that contribute to the acidity of the juices of lemon and calamansi are

also found to be present therein (Pao and Fellers, 2003). In an experiment

conducted by Kale, et al. (2012), orange juice was blended with soy milk as an

attempt to mask the unpleasant beany flavour of the latter. This attempt was

proven to be successful as not only that the orange juice had the undesirable

flavour masked, it had also enhanced the likability of the soy milk. The findings

of Kale, et al. (2012) had proven that orange juice could be useful as an

ingredient or additive in the preparation and production of foods, thus making

the utilization of orange juice as acidulant in jam making plausible and

reasonable.

Perhaps the most credible demonstration of the plausibility of applying citrus

juices as acidulants in jam making comes from the statements made by Igoe and

Hui (1996), Cheong, et al. (2012), Najafpour (2007), and Rababah, Al-u’datt

and Brewer (2015), respectively. According to Igoe and Hui (1996), and
Cheong, et al. (2012), citric acid, which is an acidulant that is commonly used

in the production of jams, are present in the fruits of lemons, calamansi as well

as oranges as the acid of predominance. Igoe and Hui (1996), and Najafpour

(2007) had also stated that this acid is extractable from, but not limited to, the

juices of lemon. On the other hand, the abundance of citric acid in the particular

juices of lemon had also been highlighted by Rababah, Al-u’datt and Brewer

(2015), and according to them, it is this attribute that leads to the frequent

incorporation of this juice in the manufacturing of jam. Though the juices of

lemon, calamansi and orange are neither the primary sources of commercial

citric acid nor the main acidulants employed in jam manufacturing, the above

statements have undoubtedly demonstrated the capability of these juices, as well

as the juices of other citrus fruits, as acidulants in jam making (Igoe and Hui,

1996; Raju and Bawa, 2006).

2.3.1.2 Exhibition of Antimicrobial Activity by Citrus Juices

The capability of citrus juices as acidulants in the making of jam has also been

demonstrated by the antibacterial activity as exhibited by the juices. Firstly, as

a citrus juice that is known for its acidity, lemon juice has had its antimicrobial

properties assessed and verified in multiple researches conducted by different

groups of scientists. In an experiment that was conducted by Tomotake, et al.

(2006), lemon juice was found to be inhibitory against seven Vibrio strains,

including three different strains of Vibrio cholerae. Based on their findings,

Tomotake, et al. (2006) attributed the antibacterial properties of lemon juice to

the citric acid present therein, saying that this acid inhibited microbial growth
via a mode of action that is the same as that of the commercial, synthetic

acidulants. On the other hand, the antibacterial properties of lemon juice was

evaluated and compared against those of orange juice in a research carried out

by Aburowais, Banu and Nisha (2017). In their research, Aburowais, Banu and

Nisha (2017) discovered that the antibacterial activity of lemon juice was

evident and superior as compared to that of orange juice, as the former was

found to be effective in inhibiting the growth of five of the six microbial strains

tested, including Clostridium spp., which is the predominant pathogenic

microbe of concern in canned foods, while orange juice was shown to be non-

inhibitory against all of the microbial strains.

The findings of Aburowais, Banu and Nisha (2017) on the antibacterial

properties of orange juice were contradicted by the results of an experiment that

was conducted by Mothershaw and Jaffer (2004), which involved the

assessment of orange juice’s antimicrobial activity against a number of spoilage

and pathogenic microbes that included Escherichia coli as well as

Staphylococcus aureus. The results of this experiment verified the antimicrobial

properties of orange juice as the juice was shown to be inhibitory against all of

the microbial species tested. Based on the results of their experiment,

Mothershaw and Jaffer (2004) inferred that the inhibitory effect of orange juice

against the microbes was contributed by the low pH of the juice, and that orange

juice’s antibacterial activity was heavily dependent upon its pH. While the

influence of pH on the antimicrobial activity of orange juice might be able to

explain the contradiction between the results obtained by Aburowais, Banu and
Nisha (2017), and Mothershaw and Jaffer (2004), the antibacterial activity of

orange juice certainly needs further verification, indeed.

On the other hand, the antibacterial activity of calamansi juice had also been

evaluated indirectly in a study conducted by Chew, et al. (2018). In their

research, calamansi juice was added to juice of sugarcane as an attempt to

prolong the shelf life of the latter by improving its microbiological stability.

This measure of improving the microbiological stability of sugarcane juice was

proven to be effective as the addition of calamansi juice was found to be

coincided with the decrease in the microbial load of sugarcane juice. On top of

that, it had also been discovered that the extent of decrement in both the total

plate count as well as the yeast and mould count of sugarcane juice was directly

proportional to the concentration of the calamansi juice added, thereby verifying

the inhibitory effect of calamansi juice against microbial growth as well as

growth of yeast and mould (Chew, et al., 2018).

2.4 Dragon Fruit

2.4.1 Nutritional and Functional Properties of Dragon Fruit

A number of health benefits have been associated with dragon fruit owing to

the nutritional and functional components that are found in the flesh and seed

of the fruit, as well as their bioavailability. From nutritional perspective, dragon

fruit is perhaps best known as a rich source of minerals, with substances such
as sodium, magnesium, potassium as well as phosphorus present in the fruit in

amounts that are remarkable and are apparently greater than those of other

tropical fruits including pineapple and mango (Liaotrakoon, 2013). Dragon fruit

is also rich in certain vitamins like vitamins C and B3, despite other vitamins,

namely vitamin A, vitamin B1 and vitamin B2, are found in the fruit in rather

small amounts. In addition, according to Wichienchot, et al. (2010 cited in

Liaotrakoon, 2013, p.13), some fructooligosaccharides are suggested to be

making up part of the oligosaccharides that exist in the dragon fruit, and these

fructooligosaccharides possess prebiotic properties and are good to the

gastrointestinal tract of human beings.

The seeds of the dragon fruit are also an undeniable contributor to the nutritional

value of the fruit as well. The presence of vitamin E and essential fatty acids in

these edible seeds in considerable amount have given rise to seed oil that

contains properties that are beneficial to human health. Besides, the seeds of

dragon fruit may also contain insoluble fibre such as cellulose and

hemicellulose, and these substances confer a number of health benefits to

humans as well (Tarpila, et al., 2005 cited in Liaotrakoon, 2013, p.14).

Apart from its nutritional value, dragon fruit has also been known to be

possessing several functional properties, most famously its antioxidative

activity. The high level of such activity as exhibited by dragon fruit is

considerably greater than those in other tropical fruits including lychee and

papaya. This is owing to the presence of abundant amounts of phenolic acids,

which have the potential of functioning as reducing agents, quenchers of singlet

oxygen and metal chelators, in dragon fruit (Liaokratoon, 2013). On the other

hand, it has also been discovered that the red-fleshed dragon fruit variety

exhibits an even greater antioxidative activity as compared to that of its white-

fleshed counterpart. This can be explained by the antioxidative properties of the

pigment betacyanin (Liaokratoon, 2013). Regardless of the varieties, the

antioxidative activity of dragon fruit is also attributed to other naturally

occurring antioxidative components that include flavonoids, tocopherol and

vitamin C, each of which being present in the fruit in high concentrations as

well (Liaokratoon, 2013).

Other than the antioxidative activity, dragon fruit is said to be exhibiting anti-

proliferative activity as well (Kim, et al., 2011 cited in Liaokratoon, 2013, p.20).

The phytochemical components that exist in dragon fruit can bring

chemoprevention, cancer cell growth inhibition, antimicrobial and anti-

inflammatory effects on cells. With that, the risk of degenerative neurological

disorder, inflammation, damage on the liver, as well as diseases and cancers

concerning the cardiovascular system can possibly be lowered with the

consumption of dragon fruit (Liaokratoon, 2013).

2.4.2 Commercial Significance of Dragon Fruit

Whilst dragon fruit has been popular in Asia as well as South America, it was a

relative unknown for Europeans and North Americans until mid-1990s (Le

Bellec and Vaillant, 2011). Although it is still being labelled as a niche product,
exports of dragon fruit from the producer countries have seen a considerable

increase in recent years, and now the fruit has earned a place in the retail of rare

exotic fruits (Le Bellec, Vaillant and Imbert, 2006). Whilst the implementation

of commercial policies in some countries from which dragon fruits are produced

and exported has been the driving force behind the success of dragon fruit in the

market of exotic fruits, the inherent qualities and attributes of the fruit is equally

important for it to gain its current status (Le Bellec and Vaillant, 2011).

At the present time, there are two main segments in the market that the dragon

fruit attracts, namely Asian and European customers. Asian customers’ demand

for the fruit is rather regular along the year, and the trade of dragon fruit in this

market segment usually peaks at the Spring Festival (Le Bellec, Vaillant and

Imbert, 2006). Conversely, European customers would not purchase dragon

fruits on a regular basis throughout the year. Instead, the purchase of dragon

fruit by these customers would primarily take place during the Christmas season,

which is also when dragon fruit, along with other exotic fruits, is popularized

(Le Bellec, Vaillant and Imbert, 2006).

2.4.3 Justification for the Processing of Dragon Fruit into Jams

The making of dragon fruit jam is primarily rationalized by the susceptibility of

the fruit towards deterioration at ambient temperature. According to Le Bellec,

Vaillant and Imbert (2006), dragon fruit has a rather short shelf life at this

temperature range, where it is able to remain fit for human consumption for only

3 to 4 days. Despite the shelf life of dragon fruit can be improved by subjecting
the fruit to storage at low temperature, rapid deterioration of the eating quality

of the dragon fruit would inevitably take place once the fruit is moved out of

cold storage (Nerd, et al., 1999 cited in Le Bellec and Vaillant, 2011, p.263).

This has been proven by Nerd, et al. (1999, cited in Mercado-Silva, 2018, p.344),

who reported an abrupt decrease in malic acid content as well as a great decrease

of firmness in the dragon fruit, which was previously stored at 6 ºC, once the

fruit was removed from storage and exposed to ambient conditions. The

increased susceptibility of the dragon fruit in the environmental conditions can

be tackled by having the fruit processed into jams, which as a means of fruit

preservation, can prolong the shelf life of the fruit and maintain its eating quality

for a greater period of time at ambient temperature, thus justifying the making

of dragon fruit jam.

 CHAPTER 3

MATERIALS AND METHODS

3.1 Materials

3.1.1 Ingredients

The ingredients utilized for this study had been classified into three categories,

namely fruits, sweetener and thickener. Whilst all of the fruits were purchased

from a local market in Kampar, Malaysia, sweetener and thickener were

obtained from the hypermarket TESCO and Siti Bakery Ingredients House

respectively, both of which being located in Kampar, Malaysia as well. The

details of the ingredients are shown in Table 3.1.

Table 3.1: List of ingredients utilized in this study.
Ingredient Category Brand Country

Dragon fruit Fruits - Malaysia

(Hylocereus

polyrhizus

(Web.) Britton &

Rose)

Orange (Citrus Fruits - Malaysia

sinensis (L.)

Osbeck)

Lemon (Citrus Fruits - Malaysia

limon (L.)

Burm.)

Calamansi Fruits - Malaysia

(Citrus

microcarpa)

Fine granulated Sweetener Gula Prai Malaysia

sugar

Pectin powder Thickener - Malaysia

3.1.2 Instruments and Materials

The instruments and microbiological materials employed in this study were

outlined in Table 3.2 and Table 3.3, respectively.

Table 3.2: List of instruments employed in this study.
Instrument Model Brand Country

Analytical ML304T Mettler Toledo USA

balance

Colorimeter CM-600D Konica Minolta Japan

Incubator IN260 Memmert Germany

Induction cooker FCC Faber Malaysia

FORNELLO

2000

Laminar flow AHC-401 ESCO Singapore

cabinet

Moisture MX-50 A&D Japan

analyser

pH meter FP20 Mettler Toledo USA

Refractometer PAL-3 ATAGO Japan

Stomacher BagMixer® 400 INTERSCIENCE France

Texture analyser TA.XT Plus Stable Micro UK

Systems

Viscometer DV2T Brookfield UK

Table 3.3: List of microbiological media utilized in this study.

Microbiological media Brand Country

Plate count agar Friendemann Schmidt USA

Ringer’s solution Merck Germany

3.2 Methodology

3.2.1 Preparation of Dragon Fruit Jam Treatments

Three treatments of dragon fruit jam, each of which differs in the citrus juice

used, were prepared according to the formulations shown in Table 3.4.

Table 3.4: Formulations of dragon fruit jam.

Treatment A B C

Ingredient (Weight, g)

Calamansi juice 5.0 - -

Lemon juice - 5.0 -

Orange juice - - 5.0

Dragon fruit 250.0 250.0 250.0

Fine granulated 125.0 125.0 125.0

sugar

Pectin 2.5 2.5 2.5

The dragon fruit was processed into jam in accordance with the procedure as

described by Mohamad, Saheed and Jamal (2012), with certain modifications.

Firstly, the cut and sliced pulp of dragon fruit was simmered over medium heat

by means of induction cooker, with the cooking power set to be 500 W. After

simmering had been allowed to persist for 5 min, fine granulated sugar was

added and the mixture was mixed well. Concurrently, the cooking power of the

induction cooker was increased to 800 W so as to allow the mixture to be

brought to boil more rapidly. Once boiling had occurred, the mixture was left to

boil for 5 min before the particular citrus juice (calamansi/orange/lemon juice)

was added. Subsequently, the mixture was allowed to boil for another 5 min,

and 2.5 g of pectin powder was added into the mixture. Simultaneously, the

cooking power of the induction cooker was reduced to 500 W. The mixture was

stirred well and finally, the jam produced was hot filled into a Mason jar that

had been sterilized prior. The jam sample was stored at room temperature until

the conduction of the analyses.

3.2.2 Physicochemical Analysis

3.2.2.1 Measurement of Colour

Colour measurement was conducted according to the procedure reported by

Basu and Shivhare (2012), such that the colour of each jam sample was

determined via a colorimeter that was calibrated with standard white calibration

plate prior to the measurement. Illuminant D65 and 10°viewing angle were

employed as the references for the colorimeter, which would measure colour by

CIELAB colour space. For each jam sample, an adequate amount of the sample

was placed in a Petri dish such that the dish was completely filled with the

sample. The values of L*, a* and b*, which represent lightness, redness to

greenness, and yellowness to blueness, respectively, were taken by the

colorimeter for each sample. The colour measurement of each jam treatment

was conducted in triplicate and the results were expressed as mean ± standard

deviation.
3.2.2.2 Measurement of Moisture Content

Moisture content of each jam sample was measured by means of moisture

analyser. For each jam sample, 2.0 g of the sample was spread evenly on a glass

fibre sheet, which was then placed on a disposable aluminium foil pan. The

aluminium foil pan, together with the content within, was placed onto the

sample pan of the moisture analyser. Quick mode operation was selected as the

measurement method used by the moisture analyser for this analysis, whereby

each sample would be subjected to heating at 200°C for about 3 min before the

drying temperature would be lowered to the temperature that was set for the

program, which was 160°C (A&D Company, 2004). The measurement would

be completed once the change in moisture content of the sample in 1 min, as

detected by the moisture analyser, had reached 0.10%. The measurement of

moisture content for each jam treatment was completed in triplicate and the

results were expressed as mean ±standard deviation.

3.2.2.3 Measurement of pH

The pH value of each jam sample was measured with a pH meter. Before

measurement, calibration of the pH meter was done using buffer solutions of

pH 4.0, 7.0 and 9.0. The pH value of each sample was obtained by having the

electrode dipped into the sample after it was rinsed with distilled water and

wiped clean. The pH measurement was completed in triplicate for each jam

treatment and the results were expressed as mean ±standard deviation.

3.2.2.4 Texture Analysis

Texture of each sample was analysed by the texture analyser. A cylindrical

probe of 36 mm in diameter was employed for the analysis. The jam sample, as

contained in a jar, was placed on a heavy duty platform. The sample was then

subjected to a compression test, whereby the cylindrical probe would be

compressing the sample at a velocity of 1.0 mm/s. Once the probe had

compressed the sample for a distance of 8 mm, it would then stop the

compression and remain stationary for 30 s, before commencing decompression.

The texture of each sample was expressed as its firmness. For each jam

treatment, texture analysis was conducted in triplicate and the results were

expressed as mean ±standard deviation.

3.2.2.5 Measurement of Total Soluble Solids (TSS)

The total soluble solids of each jam sample was measured via a refractometer,

which was calibrated with distilled water before each measurement was taken.

An adequate amount of jam sample was placed on the prism of the refractometer

such that the prism was completely covered by the sample. The total soluble

solids of the sample would be measured in °Brix by the refractometer. The

measurement of total soluble solids for each jam treatment was conducted in

triplicate and the results were expressed as mean ± standard deviation.

3.2.2.6 Measurement of Viscosity

Viscosity of each jam sample was determined using a viscometer. Spindle

number 6 was employed for the measurement. The measurement was done by

having the spindle submerged in the sample and allowing it to rotate at a rotating

speed of 5 rpm for 1 min. The viscosity of the sample would then be calculated

by the viscometer. The viscosity measurement for each jam treatment was

completed in triplicate and the results were expressed as mean ± standard

deviation.

3.2.3 Sensory Evaluation

Sensory evaluation was conducted in accordance to the procedure described by

Olivera and Salvadori (2009 cited in Tan, 2017, p.37). The sensory test

employed for this sensory evaluation was acceptance test, whereby 75 untrained

panellists from Universiti Tunku Abdul Rahman were recruited and required to

evaluate the acceptability of each jam treatment in terms of appearance, aroma,

taste, mouthfeel and overall acceptability on a 9-point hedonic scale, in which

each of the nine verbal responses was assigned with a number, with ‘9’ being

assigned to the response of ‘Like extremely’, ‘1’ being assigned to the response

of ‘Dislike extremely’, and a central point of ‘5’ that represented ‘Neither like

nor dislike’ (Olivera and Salvadori, 2009 cited in Tan, 2017, p.37; Wichchukit

and O’Mahony, 2014). Each of the jam samples served was assigned with a

three-digit random code and the samples were served in random order based on

the master sheet constructed prior. The panellists were mandated to rinse their

mouths with the rinsing water provided before evaluating each sample. The
sensory test was conducted in a sensory evaluation laboratory for interference

avoidance, and the results of sensory evaluation were expressed as mean ±

standard deviation (Olivera and Salvadori, 2009 cited in Tan, 2017, p.37).

3.2.4 Microbiological Analysis

3.2.4.1 Determination of Total Plate Count (TPC)

Total plate count of each jam sample was determined throughout a storage

period of three weeks where the samples were stored at room temperature

(Gómez, et al., 2013). The analysis of total plate count was conducted based on

the pour plate method as described by Maturin and Peeler (2001), with slight

modifications. A 10 -1 homogenate of each sample was prepared by mixing 25 g

of the sample with 225 mL of Ringer’s solution and subjecting the mixture to

homogenization by means of stomacher. Subsequently, serial dilutions of 10 -2

and 10-3 were prepared from the homogenate. One mL of each prepared dilution

was then transferred to two empty, sterile Petri plates, before 15-20 mL of

cooled molten plate count agar was added to each plate. Thorough and uniform

mixing of the mixture was done by rotating each plate in alternate directions,

and subjecting the plates to back-and-forth motion. The agar was allowed to

solidify before the plates were incubated at 37°C for 48 h. Following incubation,

the number of colonies observed on each plate was counted. Only plates with

25-250 colonies were taken into consideration, whereas counts were recorded

to be too few to count (TFTC) and too numerous to count (TNTC) for plates

with less than 25 colonies or more than 250 colonies, respectively. Total colony
forming units per gram of jam sample (cfu/g) was calculated for each jam

treatment and the results were expressed as mean ±standard deviation.

3.2.5 Statistical Analysis

The data obtained from each analysis were subjected to statistical analysis using

IBM SPSS Statistics software (Version 25). One-way analysis of variance

(ANOVA) was conducted among the data for each analysis to determine the

presence of significant difference among the mean values of the jam treatments

at significance level of p ≤ 0.05, and for the mean values in which significant

difference was found to be existing, Fisher’s Least Square Difference (LSD)

test was conducted to identify the pair(s) of mean values that is significantly

different.
 CHAPTER 4

RESULTS

4.1 Physicochemical Analysis

The results of physicochemical analysis that had been conducted on the samples

of the three jam treatments, as expressed in mean ± standard deviation of the

triplicate measurements that had been done, are shown in Table 4.1. The three

different jam treatments were shown to be possessing a rather similar colour,

moisture content, texture and total soluble solids content, with no significant

difference existing among the mean values of these three treatments as obtained

for these physicochemical parameters. As for pH, Treatment B, which used

orange juice as acidulant, had been found to acquire a distinctive and markedly

higher pH value. On top of that, in terms of viscosity, the three jam treatments

differed from each other significantly, with Treatment B (Dragon fruit jam made

with orange juice as acidulant) and Treatment C (Dragon fruit jam made with

lemon juice as acidulant) obtaining the highest and lowest mean values of

viscosity, respectively, for their samples.

Table 4.1: Mean values obtained for the jam treatments on the physicochemical
parameters measured.
Treatment A B C

Colour

L* 29.31a ±0.64 29.74a ±0.56 29.69a ±0.26

a* 0.93a ±0.26 0.75a ±0.20 0.95a ±0.30

b* 2.43a ±0.22 2.22a ±0.34 2.27a ±0.27

Moisture 36.27a ±2.70 38.44a ±0.69 35.06a ±4.63

content (%)

pH 4.26b ±0.50 4.55a ±0.35 4.32b ±0.95

Firmness (N) 1.33a ±0.30 1.09a ±0.22 1.35a ±0.28

Total soluble 77.10a ±1.22 74.77a ±1.27 76.07a ±2.32

solids (ºBrix)

Viscosity 92900.00b ± 110933.33a ± 73333.33c ±

(mPa·s) 6199.19 6123.18 1665.33

A: Dragon fruit jam made with calamansi juice as the acidulant

B: Dragon fruit jam made with orange juice as the acidulant
C: Dragon fruit jam made with lemon juice as the acidulant
a-b
: Mean values in the same row that were assigned with different superscripts
are significantly different (p ≤ 0.05).

4.2 Sensory Evaluation

The sensory scores obtained for the jam treatments with regards to their aroma,

appearance, taste, mouthfeel and overall acceptability had been expressed as

mean ±standard deviation of the scores given by 75 panellists and are presented

in Table 4.2. The three jam treatments had shown no significant difference in

the degree to which their aroma is liked, with mean scores ranging from 5.57 to
5.80. Likewise, the absence of significant difference was also recorded in the

acceptability of the jam treatments with respect to their appearance and

mouthfeel, where all of these treatments were indicated to possess a look and a

texture that were in overall, liked moderately and slightly by the panellists,

respectively. Apart from that, the taste of Treatment A (Dragon fruit jam made

with calamansi juice as acidulant) had been found to be significantly more well

received as compared to that of Treatment B (Dragon fruit jam made with

orange juice as acidulant), whereas the acceptability on taste of Treatment C

(Dragon fruit jam made with lemon juice) was not significantly different from

the previous two treatments. Similar trend had also been observed on the overall

acceptability among the jam treatments. Whilst the acceptability of the three

jam treatments on the various sensory parameters evaluated had been positive,

it is noteworthy that Treatment A (Dragon fruit jam made with calamansi juice

as acidulant) had scored a relatively higher mean values in all attributes than the

other two treatments, whereas Treatment B, on the other hand, had been

recorded with the lowest mean scores among the jam treatments.
Table 4.2: Mean sensory scores of the jam treatments with regards to the
sensory parameters evaluated.
Treatment A B C

Aroma 5.80a ±1.30 5.57a ±1.22 5.57a ±1.15

Appearance 7.12a ±1.22 6.77a ±1.51 6.85a ±1.27

Taste 6.64a ±1.54 5.81b ±1.81 6.32ab ±1.59

Mouthfeel 6.41a ±1.37 6.00a ±1.68 6.12a ±1.41

Overall 6.83a ±1.33 6.07b ±1.67 6.48ab ±1.33

acceptability

A: Dragon fruit jam made with calamansi juice as the acidulant

B: Dragon fruit jam made with orange juice as the acidulant
C: Dragon fruit jam made with lemon juice as the acidulant
a-c
: Mean values in the same row that were assigned with different superscripts
are significantly different (p ≤ 0.05).

4.3 Microbiological Analysis

Table 4.3 showed the total plate count (cfu/g) of samples of the three jam

treatments throughout three weeks of storage at room temperature. From the

table, it is indicated that the samples, regardless of the period of storage, had

been constantly giving rise to plates with less than 25 colonies, and such

phenomenon was observed in all of the three jam treatments.

Table 4.3: Total plate count (cfu/g) of samples of the jam treatments after 0, 1,
2, and 3 weeks of storage.
Week of storage Treatment

A B C

0 TFTCa TFTCa TFTCa

1 TFTCa TFTCa TFTCa

2 TFTCa TFTCa TFTCa

3 TFTCa TFTCa TFTCa

A: Dragon fruit jam made with calamansi juice as the acidulant

B: Dragon fruit jam made with orange juice as the acidulant
C: Dragon fruit jam made with lemon juice as the acidulant
TFTCa: Too few to count in 25 g
 CHAPTER 5

DISCUSSION

5.1 Physicochemical Analysis

A number of trends had been observed from the results of the physicochemical

analysis conducted against the three jam treatments. Firstly, the analysis of the

colour of each of the three treatments had shown that none of the treatments had

a rather distinguishable colour as compared to each other, as the L*, a* and b*

values obtained for the three treatments were found to be lacking of significant

difference. Despite the absence of significant difference is logical and could be

explained by plausible theory, the obtained values of L*, a* and b* for the jam

treatments, however, are perceived to be unexpected and even uncanny when

they are compared against those obtained for the pulp of different varieties of

dragon fruit as conducted by Chemah, et al. (2011). As the samples of all three

jam treatments employed red-fleshed dragon fruits, they were expected, and did

possess a colour that did not deviate much from the original dragon fruit pulp

colour. In other words, they were supposed to have L*, a* and b* values that

match those of the red-fleshed dragon fruit pulp. Whilst the L* values of the

treatments were indeed close to the L* value of the pulp of the red dragon fruit

variety (Hylocereus polyrhizus), the a* and b* values, however, are found to be

more alike to those of the pulp of white-fleshed dragon fruit (Hylocereus

undatus), in accordance with the results obtained from the experiment executed

by Chemah, et al. (2011). The only plausible reason behind this phenomenon is
that the seeds present in the dragon fruit jam might have been widely dispersed

over the Petri dish to which the samples were added and the readings were taken,

thus causing the colorimeter to measure the colour of the seeds themselves

rather than the true colour of the jam sample. This is supported by the fact that

the L* values of the three treatments, which indicate the lightness of the colour

detected and were ranging from 29.31 to 29.74, are not even close to the L*

value of the white dragon fruit pulp, which was 54.9 as according to the results

obtained by Chemah, et al. (2011). The lower L* value of the samples have also

indicated that the colour detected is much darker than that of the pulp of H.

undatus, thus reinforcing the speculation proposed.

Whilst the values of a* and b* of the three jam treatments were out of

expectations, it is deduced that all of the citrus juices used as the acidulant for

the dragon fruit jam did give rise to samples with a colour that is similar to the

pulp of the red-fleshed dragon fruit, judging from the similarity between the L*

values of the treatments and that of the red-fleshed dragon fruit pulp, as well as

the visual appearance of the samples. The retention of the colour of the pulp in

the jam samples could be attributed to the ability of each of the citrus juices to

improve the stability of betacyanin, which is the predominant pigment found

within the red-fleshed pitahaya’s pulp, and prevent it from degradation upon

thermal treatment (Liaotrakoon, 2013). According to Liaotrakoon (2013), citric

acid, along with ascorbic and isoascorbic acids, is able to enhance the stability

of betacyanin which would otherwise decompose and result in colour loss upon

heating. As the citrus juices are rich in citric and ascorbic acids, it is apparent
that these juices were responsible for the preservation of colour in the dragon

fruit jam samples.

Other than the values of L*, a* and b*, the three jam treatments had also shown

to be indistinguishable in terms of their moisture content. With the range of

35.06% to 38.44%, the mean moisture content obtained for the three jam

treatments is reasonable as the range does not deviate much from the range of

various types of fruit jams available on the markets in Malaysia (31.23-33.36%),

the latter of which being obtained from an experiment conducted by Mohd

Naeem, et al. (2017). On the other hand, it has also been discovered that the

moisture content of the jam samples is enormously lower than that of the dragon

fruit, which ranges from 83% to 89% according to Liaotrakoon (2013). The

moisture content of the three jam treatments is apparently a result of moisture

removal of the jam samples upon thermal treatment, as the samples were

allowed to boil for 10 min. In other words, the source of acidulant would have

no effect over the moisture content of the jam samples.

Despite there has been no significant difference over the colour and moisture

content of the three jam treatments, the different type of citrus juices used had

been proven to be influential to the pH of the jam samples. This was indicated

by the results that Treatment B (Dragon fruit jam made with orange juice as the

acidulant) had a significantly higher pH than the other two jam treatments. The

significant difference observed among the pH of the three jam treatments is

evidently resulted from the difference in acidity of the citrus fruits to which the
citrus juices were extracted. According to Bamise and Oziegbe (2013), and Lee

(2000), among the three citrus juices that were employed for the current

experiment, orange juice would be the one with the lowest acidity, and

calamansi juice, on the other hand, would be the most acidic citrus juice. This

was proven by the results of their analyses, whereby the mean pH of the juices

of oranges, lemons and calamansi were found to be 4.0, 3.1 and 2.4, respectively

(Bamise and Oziegbe, 2013; Lee, 2000). These findings are discovered to be

matching with the results obtained for the current experiment, where Treatment

B (Dragon fruit jam made with orange juice as the acidulant) was determined to

be the treatment with the highest pH, and Treatment A (Dragon fruit jam made

with calamansi juice as the acidulant) was found to have the lowest pH among

the three treatments analysed, thus justifying the positive correlation between

the acidity of the citrus juice used and that of the dragon fruit jam.

Though significant difference had been found to be present among the pH of

the three jam treatments, the results of texture analysis revealed that such

difference would have no influence over the firmness of the three jam treatments.

As the firmness of the jam treatments reflects the strength of the gel structure

formed within the samples of the jam treatments, it is apparently determined by

the extent of gel formation within the samples. According to Featherstone

(2016), the amount of pectin used would be the pivotal determinant of the

continuity of the gel structure formed, which in turn influences the gel strength.

On the other hand, the acidity, and the concentration of the sugar in the jam

would control the rigidity of the gel (Featherstone, 2016). As both the amount

of pectin and sugar employed in the formulation of the three jam treatments was
the same, the absence of significant difference among the firmness of the three

jam treatments is deemed to be logical. Conversely, whilst significant difference

was found to be existing among the pH of the jam treatments, the results of

firmness suggested that the difference in pH was not extensive enough to lead

to a distinction in the firmness of the jam treatments. Consequently, the type of

citrus juices used as the acidulant is speculated to have no notable effect on the

firmness of the dragon fruit jam.

Next, a similar trend had also been observed for the results of total soluble solids

content of the three jam treatments, where significant difference was found to

be absent. The total soluble solids content of the three jam treatments, which

ranged from 74.77% to 77.10%, is rather similar with those of the strawberry

jam analysed by Owolade, et al. (2016), the latter of which exhibited a mean

total soluble solids content of 80.00% at Day 0 of storage. This indicates that

the results of total soluble solids of the jam treatments are rather unsurprising

and understandable. According to Silva, et al. (2006 cited in Rodiah, Nurul

Aishikin and Nurul Ain, 2017, p.51), there exists a high and positive correlation

between the sugar content of a jam and its total soluble solids content.

Consequently, as the amount of sugar used in the formulation of the three jam

treatments was constant, the absence of significant difference among the total

soluble solids content of the jam treatments is rational and reasonable. Apart

from that, the obtained results have also indicated that the type of citrus juices

used as the acidulant would not pose an impact on the total soluble solids

content of the dragon fruit jam, despite Liu, Heying and Tanumihardjo (2012)

pointed out that orange juice has markedly higher sugar content compared to
lemon juice. This is likely to be owing to the amount of citrus juice added to

each jam sample. With only 5 mL of citrus juice added to each sample that was

in turn derived from 250 g of dragon fruit pulp, it is reasoned that the distinction

in the total sugar content of the citrus juice would be too minimal to lead to a

difference in the total sugar content and ultimately, the total soluble solids

content of the three jam treatments.

Finally, the results of the viscosity of the jam treatments revealed that the three

different citrus juices used as the acidulant for the jam treatments had given rise

to dragon fruit jam with viscosity that was distinctive from one another.

Whereas Treatment C (Dragon fruit jam made with lemon juice as the acidulant)

did have a viscosity (73333.33 mPa·s) that is within the viscosity range

exhibited by the commercial jam samples examined by Jaiswal, Patel and Naik

(2015), which was 36820-78980 mPa·s, Treatment A (Dragon fruit jam made

with calamansi juice as the acidulant) and Treatment B (Dragon fruit jam made

with orange juice as the acidulant) had a noticeably higher viscosity compared

to those of the commercial jam samples. According to Bourne (2002), the

difference in the viscosity of the jam treatments could be explained by the

difference in either the average molecular weight of the solutes present in the

jam, or the suspended matter present in the jam, or even both. If the average

molecular weight of the solutes was the sole factor that resulted in the distinction

in the viscosity among the jam treatments, it would indicate that the average

molecular weight of solutes of the dragon fruit jam made with orange juice

would be the greatest among the three jam treatments, and that lemon juice

would have given rise to dragon fruit jam with the lowest average molecular
weight of solutes, based on the relationship between viscosity and the molecular

weight of solute. Conversely, if the difference in viscosity of the jam treatments

was attributed to merely the difference in the suspended matter, it would then

infer that the dragon fruit jam with the greatest concentration of suspended

matter would be the one that employed orange juice, followed by those that were

derived from calamansi juice, and lemon juice. Unfortunately, the inferences

made could not be validated as the average molecular weight of solutes, as well

as the concentration of suspended matter of each of the citrus juices are yet to

be well established, making comparison among the different citrus juices in

such aspects not feasible at the moment. Nevertheless, the type of citrus juices

used as the acidulant has been proven by the results to be influential on the

viscosity of the dragon fruit jam.

5.2 Sensory Evaluation

Several inferences on the effect of different types of citrus juices as the

acidulants on the sensorial properties of the dragon fruit jam can be made based

on the results of the sensory evaluation conducted. First and foremost, it is

speculated that the acidulant used would not influence, impart or enhance the

aroma of the dragon fruit jam, as there is no significant difference among the

mean scores obtained for the acceptability of each jam treatment in terms of

aroma. Whilst the generalized response towards the aroma of the three jam

treatments was leaning towards neutral, there had been a considerable number

of panellist who commented that little or no aroma had been detected from the

served jam samples. This can be explained by the effect of heat treatment on the
aroma-active compounds of the freshly-squeezed citrus juices used as the

acidulants. According to Perez-Cacho and Rouseff (2008), fresh orange juice

that has been extracted manually is not stable, and its aroma composition would

be greatly affected if the juice is subjected to thermal processing. On top of that,

the orange juice that has received thermal processing would have its flavour

negatively affected by the process owing to the reduction in the concentration

of the original volatile compounds in the juice as induced by the heat treatment

(Bazemore, Goodner and Rouseff, 1999 cited in Perez-Cacho and Rouseff, 2008,

p.9785). Considering that the jams were allowed to boil for another 5 min after

the addition of the citrus juices as the acidulant, the heat treatment had likely

resulted in the weak or absence of aroma in the final dragon fruit jam samples.

In addition to the aroma of the three jam treatments, the panellists had no

preference on any of the jam samples served in terms of their appearance. The

obtained results for the acceptability of the appearance of the three jam

treatments indicated that every treatment had received a mean score that is close

to 7 (Like moderately), and there is no significant difference between these

mean scores. As Leon, et al. (2006 cited in Tanner, 2016, p.2) stated that the

surface colour of a food would be the principal determinant of its appearance, it

was inferred that the acceptability of the appearance of the jam treatments was

attributed to the natural colours of the dragon fruit pulp. Consequently, the

acidulants should also be accredited for the results owing to their function of

colour preservation. On the other hand, the absence of significant difference as

observed in both the mean scores of the acceptability in appearance and the

values of L*, a* and b* obtained by the three jam treatments has also reinforced
the proposition that the three citrus juices added to the dragon fruit jam had

exhibited similar extent of colour retention.

As for the acceptability in terms of taste, significant difference had been found

to be existing among the mean scores obtained for the three jam treatments. The

taste of Treatment A (Dragon fruit jam made with calamansi juice as the

acidulant) was indicated to be better received than Treatment B (Dragon fruit

jam made with orange juice as the acidulant), and the acceptability of the taste

of Treatment C (Dragon fruit jam made with lemon juice as the acidulant) was

found to be indistinguishable from those of the previous two treatments. On top

of that, a number of panellists had also remarked that the jam samples served

were considerably too sweet to be evaluated alone without the use of bread as

carrier. This leads to the speculation that the significant difference obtained for

the acceptability of the taste of the three jam treatments is resulted from the

differing extent to which the sweetness of the dragon fruit jam was cut by the

citrus juices added. According to Agam (1994), acidulants are essential for the

balancing of the excessive sweetness as imparted by the sugar as they act by

supplying a “sour” taste to the food. The “sour” taste, on the other hand, is

dependent upon the pH and the organic acid content of the acidulants (Da

Conceicao Neta, et al., 2007). Whilst the organic acid content of the three jam

treatments had not been analysed, the results of the analysis of pH do support

the inference made, as Treatment A did have a significantly lower pH compared

to Treatment B. This would mean that calamansi juice (Treatment A) was able

to impart a greater sourness to the dragon fruit jam and hence, reduce the

sweetness of the sugar to a greater extent compared to orange juice. On the other
hand, the pH of Treatment C was found to be significantly different from

Treatment B, however the acceptability of taste of these two treatments is not

distinguishable from each other. Considering that both the pH and acceptability

of taste of Treatment C was also not significantly different from those of

Treatment A, the phenomenon could have been attributed to the differing

organic acid profile of each of the citrus juice used. Nonetheless, the correlation

between the sourness imparted by the used acidulant and the acceptability of the

taste of the dragon fruit jam infers that the panellists had a preference on

whichever jam sample that was less sweeter than the others, thus explaining the

result that Treatment A was preferred over Treatment B.

Whereas the taste of the three jam treatments were found to be liked to a

different degree, their acceptability in terms of mouthfeel had been proven

otherwise. All jam treatments evaluated had received a mean score that is close

to 6, showing that the mouthfeel of each of these jam treatments was liked

slightly by the panellists. The results obtained for the acceptability of mouthfeel

of the jam treatments were apparently attributed to the structure and properties

of the gel formed therein, which in turn were influenced by the acidulant in

conjunction with sugar and pectin. Whilst the results obtained for the

acceptability in mouthfeel of the three jam treatments match that of the firmness

of the jam treatments, such that significant difference was found to be absent in

both, an opposite trend is observed when the former is compared against the

results of the viscosity of the jam treatments, to which significant difference had

been found to be existing. Such contradiction would suggest that the difference
in the viscosity of the jam treatments did not influence the acceptability of the

mouthfeel of these jam treatments.

Last but not least, the results obtained for the overall acceptability of the jam

treatments indicated that Treatment A (Dragon fruit jam made with calamansi

juice) was significantly better-liked compared to Treatment B (Dragon fruit jam

made with orange juice) in overall, and Treatment C (Dragon fruit jam made

with lemon juice) had been found to be indistinguishable from the other

treatments. Considering that the results obtained for overall acceptability of the

jam treatments are rather similar in trend to that of the taste of the jam treatments,

and significant difference was found to be absent among all the other sensory

attributes evaluated, it was apparent that the overall acceptability of the jam

treatments was directly influenced by the acceptability in taste of these jam

treatments.

5.3 Microbiological Analysis

The results of the microbiological analysis conducted against the samples of the

three jam treatments have indicated that the dragon fruit jam, regardless of the

type of citrus juices used as the acidulant, would be microbiologically stable for

at least three weeks following its preparation, when stored under room

temperature. These results were indeed expected, considering the acidification

effect of the different citrus juices as well as the synergistic antimicrobial effect

of pH by the acidulants and the heat treatment that the jam samples had received.
In the first round of microbiological examination, all the samples of the three

jam treatments, which were prepared 1 day prior to the analysis, had given rise

to plates with less than 25 colonies. This result was mainly resulted from the

heat treatment that the jam samples received and the employed technique of hot

filling during the transfer and storage of jam samples in Mason jars, as

suggested by McGlynn (2016).

Besides, the acidulant used in each jam treatment is also a critical cause as well.

All of the pH of the final jam samples were 4.55 or below, thus making the

dragon fruit jam a high-acid food possessing a pH value that is not greater than

4.6 (McGlynn, 2016). Being a high-acid food would mean the dragon fruit jam

does not require a severe heat treatment that would be compulsory for those

foods with pH of greater than 4.6. Instead, the heat treatment of the dragon fruit

jam would only mandate the sample to reach a certain temperature of

pasteurization that is recommended based on its pH. According to McGlynn

(2016), the temperature of 99°C is the highest pasteurization temperature among

the temperatures recommended, all of which being adequate to be lethal to all

of the viable microorganisms present in the food matrix. Therefore, since each

of the jam samples had been heated to boiling, which is certainly at a

temperature greater than 99°C, and allowed to boil for 10 min upon its

preparation, the results for the first round of microbiological analysis was

completely reasonable.
The samples of the three jam treatments, which had been stored under room

temperature, would continue to give rise to plates with fewer than 25 colonies

for the subsequent rounds of microbiological analysis. This is understandable

as the dragon fruit jam samples had been well preserved by means of pH control

via the acidulants in addition to the control of water activity by the sugar

incorporated, despite Lücke (2003) stated that the employment of acidulants in

foods is often associated with objectives other than to achieve microbial

stability in these foods. Whilst citric acid and malic acid, as the predominant

organic acids in citrus juices, are not lipophilic and thus, are not capable of

equilibrating across the cell membrane of the microbes in the absence of specific

carrier proteins and inhibiting the microorganisms via cytoplasm acidification,

these acids would nonetheless be effective in microbial inhibition by means of

establishing a low pH environment (Lücke, 2003). According to Booth and

Kroll (1985 cited in Shafiur Rahman, 2007, p.291), by lowering the pH of the

dragon fruit jam, the acidulants would be able to limit the rate of microbial

growth by inactivating at least one indispensable enzyme activity that occurs on

the surface layers of microbial cells, and obstructing the uptake and excretion

of ions and nutrients by the microorganisms. This justifies the observed

antimicrobial effect of the citrus juices used as the acidulant for each of the three

jam treatments.

The results of the microbiological examination are found to be consistent with

that of a similar analysis conducted by Rodiah, Nurul Aishikin and Nurul Ain

(2017) against their newly developed banana jam. However, their research had

incorporated sodium benzoate, a preservative, into their banana jam formulation

and hence, the microbial stability of their banana jam was attributed to

predominantly the action of this preservative rather than the acidulant, though

the antimicrobial effect of the latter could be critical as well. This in turn

suggests that the dragon fruit jam must have not been preserved by solely pH

control via the acidulant, otherwise it would not be as microbiologically stable

as the banana jam developed by them.

According to Raju and Bawa (2006), the dragon fruit jam might have also been

protected by the chelating action of citric acid towards essential minerals, which

poses a secondary inhibitory effect apart from acidification towards the

microorganisms, as these compounds are vital for their growth. They also

argued that the chelating action of citric acid on essential metal ions would be

the primary mechanism behind the acid’s antimicrobial effect, rather than its

ability to lower the pH of the food.

On the other hand, Steward-Woods (2017) discovered that the lemon juice acts

on the bacteria by a method that is distinctive from acidity in his research on the

lemon juice’s antimicrobial effect towards Acinetobacter baumannii. Instead, it

would affect the bacteria by interfering with the RNA synthesis within these

microorganisms. Other than that, the flavonoids present in the lemon juice are

also accountable for its antimicrobial effect (Steward-Woods, 2017). Given the

similarity of the three citrus juices used in the current research, these findings

of Steward-Woods (2017) could be able to explain the antimicrobial effect of

the citrus juices and ultimately, the microbial stability of the three dragon fruit

jam treatments.

5.4 Recommendations for Future Research

Several types of analyses can be taken into consideration for future research in

order to investigate the effect of different type of citrus juices as acidulants on

the properties of the dragon fruit jam. Firstly, determination of the yeast and

mould count of the jam treatments can be carried out to further reinforce the

interpretations and the results of the conducted microbiological analysis that all

of the citrus juices are capable of inhibiting yeast and mould growth in the

dragon fruit jam effectively for at least three weeks as well.

Next, as citrus fruits are known to be rich in biologically active phytochemicals,

as according to Liu, Heying and Tanumihardjo (2012), analyses on the

antioxidant activity of dragon fruit jam made with the three different citrus

juices as acidulants is highly recommended. Last but not least, rheological

analysis of the jam treatments is also suggested so that a more comprehensive

understanding can be gained on the effect of different type of citrus juices used

as the acidulant on the rheological properties of the dragon fruit jam.

 CHAPTER 6

CONCLUSION

In conclusion, a number of similarities and differences have been found among

the physicochemical and sensorial properties, as well as the microbiological

quality of dragon fruit jam made with different type of citrus juices (orange,

lemon and calamansi) as acidulants. In terms of the physicochemical properties,

samples of dragon fruit jam, regardless of the type of citrus juices used as the

acidulant, possessed similar colours, moisture content, firmness, and total

soluble solids content. On the other hand, the type of citrus juices used was

proven to be influential to the pH of dragon fruit jam, with orange juice being

discovered to have given rise to dragon fruit jam with pH that was significantly

higher than those of the jams as derived from lemon and calamansi juices. On

top of that, it has also been found that each of the citrus juices used would give

rise to dragon fruit jam with viscosity that was distinctive from one another.

As for the sensorial properties, it has been proven that the type of citrus juices

used as the acidulant would not pose an influence of the acceptability of the

dragon fruit jam in terms of its aroma, appearance and mouthfeel. The taste and

overall acceptability of dragon fruit jam, however, was indeed dependent upon

the type of citrus juices used. It was indicated that calamansi juice, owing to its

superior acidity over orange juice, had given rise to dragon fruit jam that was

evidently better-liked by the consumer in terms of its taste and overall

acceptability. In overall, all the citrus juices used as the acidulants were able to

give rise to dragon fruit jams that were well received by the panellists in terms

of their appearance, taste, mouthfeel, and overall acceptability, though these

citrus juices were not able to impart a likable aroma in these dragon fruit jams.

Last but not least, it has also been implied that each of the citrus juices used was

able to preserve the dragon fruit jam to an extent where the jam, when stored at

room temperature, could remain microbiologically stable for at least three

weeks following its preparation. In other words, there was no significant

difference in the antimicrobial activity of the citrus juices used, as all of these

citrus juices were shown to have the ability to inhibit microbial growth and

proliferation in the dragon fruit jam for at least three weeks, even when the jam

had been stored under the condition that was in microorganisms’ favour.