COLLEGE OF AGRICULTURAL AND

ENVIRONMENTAL SCIENCES

Department of Food Technology and Nutrition

Nutritional Composition, Anti-oxidant Potential and

Shelf Stability of Mint Flavored Passion Fruit Juice
William Ondoma

16/U/18973

A report Submitted to the Department of Food Technology and Nutrition in

Partial Fulfillment of the Requirements for the Award of the Degree of
Bachelor of Science in Food Science and Technology of Makerere University

January 2021
DECLARATION

i
APPROVAL

ii
 DEDICATION

This research is dedicated to my beloved father Mr. Drileyo Jino, mother, Ms. Candiru Mary, as
well as Ms. Ndera Loyce, relatives, course mates, and friends who supported me throughout my
educational journey.

iii
 ACKNOWLEDGEMENT

I am grateful to Dr. Stellah Byakika for her guidance and assistance throughout the research
especially with regard to the microbiological analyses. Great thanks to Assoc. Prof. Ivan Muzira
Mukisa not only for his fatherly advice but also for the supervision of this research. I also thank
Mr. Emmanuel Okalany, the Principal Technician in the Food Chemistry Laboratory of the
Department of Food Technology and Nutrition, Makerere University for his technical support
during the physicochemical analyses. This research project was sponsored by Wales of Africa
Grants Scheme (WFA_Rd1_07) through the Community Enterprise Model for Plant Oil
Production (CEMPOP) U Ltd. I acknowledge CEMPOP’s Mr. John Ssebbale for all the support
towards the development of mint flavored juices. Lastly, I thank the Government of Uganda for
the scholarship offered that enabled me pursue my studies at Makerere University.

iv
 TABLE OF CONTENTS

DECLARATION ......................................................................................................................i

APPROVAL ........................................................................................................................... ii

DEDICATION ....................................................................................................................... iii

ACKNOWLEDGEMENT ....................................................................................................... iv

TABLE OF CONTENTS.......................................................................................................... v

LIST OF TABLES ................................................................................................................viii

LIST OF FIGURES ................................................................................................................. ix

ABBREVIATIONS AND ACRONYMS .................................................................................. x

ABSTRACT ........................................................................................................................... xi

1 INTRODUCTION ............................................................................................................ 1

1.1 Back ground ............................................................................................................... 1

1.2 Problem Statement...................................................................................................... 2

1.3 Overall objective ........................................................................................................ 3

1.3.1 Specific objectives ............................................................................................... 3

1.4 Hypotheses ................................................................................................................. 3

1.5 Significance ............................................................................................................... 4

2 LITERATURE REVIEW .................................................................................................. 5

2.1 Passion fruit ............................................................................................................... 5

2.1.1 Description, production and utilization ................................................................. 5

2.1.2 Passion fruit juice composition and processing ..................................................... 6

2.1.3 Passion fruit juice blends ................................................................................... 11

2.2 Mint ......................................................................................................................... 12

v
 2.2.1 Description and production ................................................................................ 12

2.2.2 Utilization of mint ............................................................................................. 13

2.2.3 Composition and use of mint oil and hydrosol .................................................... 14

2.2.4 Antioxidant properties and other health benefits of mint ..................................... 16

2.2.5 Antimicrobial properties and other effects of mint on composition, and stability of
foods 17

3 MATERIALS AND METHODS ..................................................................................... 20

3.1 Sample preparation ................................................................................................... 20

3.1.1 Raw materials .................................................................................................... 20

3.1.2 Preparation of the mint extract ........................................................................... 20

3.1.3 Preparation of mint hydrosol .............................................................................. 20

3.1.4 Preparation of the passion fruit juice................................................................... 20

3.1.5 Preparation of mint flavored passion fruit juice ................................................... 21

3.2 Determination of effect of spearmint extract and hydrosol concentration on sensory

acceptability of the prepared juices...................................................................................... 21

3.3 Determination of effect of spearmint extract and hydrosol concentration on the nutritional
composition and antioxidant property of passion fruit juice .................................................. 22

3.3.1 Nutritional Composition .................................................................................... 22

3.3.2 Anti-oxidant property ........................................................................................ 23

3.4 Determination of effect of spearmint extract and hydrosol concentration on storage

stability of passion fruit juice .............................................................................................. 23

3.4.1 Physicochemical analyses .................................................................................. 24

3.4.2 Microbial storage stability .................................................................................. 24

3.5 Data analysis ............................................................................................................ 25

4.0 RESULTS AND DISCUSSION ................................................................................... 26

vi
 4.1 Effect of spearmint extract and hydrosol concentrations on the sensory acceptability of
passion fruit juices .............................................................................................................. 26

4.2 Effect of spearmint extract and hydrosol concentration on the nutritional composition and
antioxidant property of passion fruit juice ............................................................................ 28

4.3 Effect of spearmint extract and hydrosol concentration on storage stability of passion fruit
juice 31

4.3.1 Physicochemical storage stability ....................................................................... 31

4.3.2 Microbial stability ............................................................................................. 32

5 CONCLUSIONS AND RECOMMENDATIONS ............................................................ 33

5.1 CONCLUSIONS ...................................................................................................... 33

5.2 RECOMMENDATIONS .......................................................................................... 33

REFERENCES ...................................................................................................................... 34

APPENDICES ....................................................................................................................... 48

Appendix I: Juices acceptable to consumers and selected for nutritional composition and shelf
stability studies ................................................................................................................... 48

Appendix II: Total Plate Count of the mint flavored juices ................................................... 48

Appendix III: Yeasts and Moulds of the mint flavored juices during storage ......................... 49

Appendix IV: Total coliforms in the mint flavored juices during storage .............................. 49

Appendix V ANOVA tables for sensory analysis of the mint flavored passion fruit juices .... 49

Appendix VI: ANOVA tables for nutritional composition of the mint flavored passion fruit juice
.......................................................................................................................................... 52

Appendix VII: Anova tables for physicochemical properties of the juice during storage........ 53

Appendix VII Sensory evaluation form ............................................................................... 55

vii
 LIST OF TABLES

Table 1 Proximate composition of passion fruit juice (per 100 g) ............................................... 8

Table 2: Mineral composition (per 100g dry weight) of passion fruit juices ................................ 9
Table 3 Compounds found in mint essential oil ....................................................................... 15
Table 4 Antimicrobial properties of mint ................................................................................. 18
Table 5 Formulations of mint flavored passion fruit juice ........................................................ 21

viii
 LIST OF FIGURES

Figure 1: Appearance of Commercially important species of mint ............................................ 12

Figure 2: Effect of mint extract (a) and hydrosol (b) on the sensory acceptability of the passion
fruit juice ............................................................................................................................... 26
Figure 3: Vitamin C content of the mint flavored juices ........................................................... 28
Figure 4: Effect of Mint on beta-carotene content of passion fruit juice .................................... 29
Figure 5: Effect of mint extract and hydrosol on total antioxidant content of passion fruit juice 30
Figure 6: Effect of mint extract and hydrosol on the pH (a) and titratable acidity (b) of passion fruit
juices during storage at 25°C for 140 days............................................................................... 31

ix
 ABBREVIATIONS AND ACRONYMS

SFTNB School of Food Technology Nutrition and Bio-engineering

CEMPOP Community Enterprise Model for Plant Oil Production

ME Mint Extract

EO Essential Oil

PDA Potato Dextrose Agar

PE Petroleum Ether

SDGs Sustainable Development Goals

DPPH 2,2-diphenyl-1-picrylhydrazyl

DCPIP Dichlorophenol indophenol

ANOVA Analysis of Variance

CVDs Cardio Vascular Diseases

AOX Alternative oxidase

BHA Butylated hydroxylanisole

VRBL Violet Red Bile Lactose

x
 ABSTRACT

Previously, mint has been used to flavor several foods but no emphasis was put on flavoring certain
juices such as passion fruit juice. Thus, its effect on the properties of passion fruit juice were
unknown. Much as different extracts of mint are used to improve the properties of foods, there is
scanty information on use of mint hydrosol, a by-product of mint essential oil production. This
research evaluated mint as a flavorant in passion fruit juice. Juice formulations containing 4-16%
of either mint extract (n=4) or hydrosol (n=4) were used. The effect of mint on the nutritional
composition, antioxidant potential, sensory acceptability (n=25 panelists), and shelf stability of the
fruit juice were monitored at 0, 35, 70, and 140 days of storage at room temperature (24±4°C).
Addition of mint extract or hydrosol significantly (p<0.05) influenced the sensory acceptability of
passion fruit juice. Acceptability scores reduced with increase in mint extract or hydrosol
concentration. Among the mint flavored juices, that with 4% extract or hydrosol was the most
accepted and was quite similar to the control. This was followed by juice flavored with 8% extract
or hydrosol while that with 16% was unacceptable. Nutrient composition of the juice in terms of
vitamin C, beta-carotene and total antioxidants significantly increased with increase in mint extract
concentration in the juice (p<0.05). Of the three acceptable juice formulations, that containing 8%
mint extract had the highest amount of vitamin C (7.81±0.00 mg/100ml), beta carotene
(104.81±0.43 µg/100ml), and total antioxidants (303.04±0.34 mg ascorbic acid equivalent /100
ml). The vitamin C and beta carotene of samples containing hydrosol was not significantly
different from plain passion fruit juice (p>0.05). The juice samples were stable throughout the
storage time. Passion fruit can, therefore, be flavored with up to 4% mint extract or hydrosol
without negatively affecting its sensory properties whilst improving its antioxidant potential.

xi
 1 INTRODUCTION

1.1 Back ground

Fruits are an important part of human diet and posses several nutritional and health benefits. They
are good sources of essential elements such as water, vitamins, minerals and organic compounds
very important for proper functioning of the body (Okwu & Emenike, 2006). They are great
sources of antioxidants responsible for scavenging free radicals (Kaur & Kapoor, 2001). These
benefits make them a predominant raw material for making juices rich in vitamins, minerals and
naturally occurring phytonutrients hence contributing to good health (Septembre-Malaterre,
Remize, & Poucheret, 2018). Several fruits, including passion fruits, can be used to make juices.

Passion fruit (Passiflora edulis) is grown as a fresh fruit for making products like fruit juice for
human consumption (Akpan & Kovo, 2005). Passion fruit is a nutritious fruit and juice extracted
from it is of high nutritional value and great importance to human health. It is a rich source of
vitamin A, vitamin C and contains fair amounts of iron, potassium, phosphorus and magnesium
(Thokchom & Mandal, 2017). Passion fruit juice also has anti-inflammatory, anticonvulsant,
antimicrobial, anticancer, anti-diabetic, antihypertensive, anti-sedative, antioxidant properties and
has been used for treating conditions such as osteoarthritis, asthma and act as colon cleanser (Zas
& John, 2016). These make it an important fruit for making a nutritious and healthy drink.

Several foods/flavourants can be incorporated in passion fruit juice with positive outcomes. For
instance, Sousa et al., (2010) blended passion fruit with cashew apple, papaya, guava and acerola
fruit juices as well as caffeine to produce a mixed fruit nectar. The nectar was high in vitamin C
and remained microbiologically stable during storage for 6 months at room temperature (25 °C).
However, vitamin C content significantly reduced during storage at room temperature (25 °C).
The product also had good sensory acceptance. Despite this information, the use of popular
flavourants such as mint for flavoring passion fruit juice and its effect on the juice properties is
lacking.

Mint (M. spicata L.) is a herbaceous, rhizomatous, and perennial Mentha species native to northern
England (Kuruppacharil, 2006). It is widely cultivated in tropical and temperate climates
(Lawrence, 2007; Hajlaoui, Trabelsi, Noumi, & Snoussi, 2009). Mint is largely used in the

1
pharmaceutical, perfume, cosmetic, tobacco and food industries. It has significant antimicrobial,
antioxidant (Kanatt, Chander, & Sharma, 2007), anti-inflammatory (Arumugam, Priya, Subathra,
and Ramesh., 2008), antitumor, anti-cancer and antiallergenic potential (Sato, and Tamura., 2015).
Its leaves are a good source of beta-carotene, calcium, iron and vitamin C (Majumdar, Vasudish,
Premavalli, & Bawa, 2008). The leaves also contain riboflavin and thiamin (Bose, 1985). It is an
important flavor after vanilla and citrus flavors (Arslan, Ozcan, & Mengeş, 2010). These benefits
make mint widely utilized in foods. For example it has been used in ash gourd juice (Majumdar
et al., 2008), grape juice (Poornimah & Ritu, 2018), pineapple and guava juice (Shalini & Ritu,
2008), herbal teas and traditional medicine (Lv, et al., 2012).

Much as mint has been used in many foods with positive impact on their properties, there is no
literature on its adoption as a flavour in passion fruit juice. Its effects on properties of passion fruit
juice remain unknown. This study, therefore, focused on adopting mint extract and hydrosol (by-
product of essential oil production) as a flavor for passion fruit juice and determining its effect on
consumer acceptability, nutrient content, antioxidant property and shelf stability of the fruit juice.

1.2 Problem Statement

There is an increasing occurrence of diseases like cardiovascular diseases and cancer in the world
(Mendis et al., 2011). They are mainly caused by oxidation of the body cells after exposure to
oxidative species like free radicals (Bahorun, Soobrattee, Luximon-ramma, & Aruoma, 2006).
Free radicals cause gene mutations which result into cancer (Ames & Shigenaga 1992),
development of atherosclerosis (Schachinger and Zeiher 2002), pathogenesis of atherosclerosis
(Galle et al 2006), and congestive heart failure (Mariani et al 2005). Damages induced by free
radicals are also associated with impaired glucose tolerance, β-cell dysfunction, and type 2 diabetes
mellitus (Jay et al 2006; Wright et al 2006). Halliwell (1996) indicates that harmful effects of free
radicals are balanced by the action of antioxidants, some of which are enzymes present in the body.
Vitamin C, carotenoids, and natural flavonoids are some of the non-enzymatic antioxidants
(McCall and Frei, 1999). These can be obtained from antioxidant rich foods such as fruits and
herbs like mint (Haseeb, Ghulam, & Imtiaz, 2018).

Mint contains vitamins, minerals, phenol, flavonoids and carotenoids (Bose, 1985). It has been
widely used in foods such as herbal teas (Lv, et al., 2012), and fruit juices; ash gourd (Benincasa
hispida) juice (Majumdar et al., 2008), grape juice (Poornimah & Ritu, 2018), and pineapple juice
2
and guava leaves extract (Shalini & Ritu, 2008). Despite its wide range of application in different
fruit juices, there is no literature of its utilization in passion fruit juice. There is thus limited
information on the effects of mint extracts or hydrosol on the composition and acceptability of
passion fruit juice. This study, therefore, evaluated the effect of mint extract and its hydrosol on
sensory acceptability, antioxidant potential and shelf stability of passion fruit juice.

1.3 Overall objective

To develop an acceptable, shelf stable, nutrient rich mint-flavored passion fruit juice

1.3.1 Specific objectives

1. Determine the effect of mint extract and hydrosol on the sensory acceptability of passion
fruit juice
2. Determine the effect of mint extract and hydrosol on nutritional composition and anti-
oxidant potential of passion fruit juice
3. Determine effect of mint extract and hydrosol on the storage stability of passion fruit juice

1.4 Hypotheses

1. Mint extract and hydrosol improves the sensory acceptability of passion fruit juice.
2. Addition of mint extract or hydrosol improves the nutritional composition and antioxidant
potential of passion fruit juice.
3. Addition of mint extract or hydrosol improves the storage stability of passion fruit juice.

3
1.5 Significance

The Sustainable Development Goals (SDG) of 2030; no poverty (SDG 1), no hunger (SDG 2), and
good health (SDG 3), can be achieved through production and consumption of nutritious vegetable
and fruit-based foods. Malnutrition and related diseases like cancer and cardio-vascular diseases
(CVDs) can be overcome by nutrient rich foods and with new food strategies (Thompson &
Amoroso, 2014). Cardiovascular diseases are one of leading causes of morbidity and mortality
globally (Mendis et al., 2011). Consumption of antioxidant rich foods is one of the preventive
measures which could help prevent the CVDs. Passion fruit and mint-based foods contain high
vitamins, minerals, fibre, and phenolic compounds which confer antibacterial, anticancer and
antioxidant benefits (Bardaweel et al., 2018; Kanatt, Chander, Sharma, 2005). This study,
therefore, contributes to managing the CVDs by developing an antioxidant rich mint flavored
passion fruit juice. Information is also provided on the effect of mint on sensory acceptability,
nutritional composition and shelf stability of passion fruit juice. This information can be utilized
by food processors and researchers in formulating mint flavored fruit products. Mint flavored
passion fruit juice may, therefore, be helpful in managing hidden hunger and cardiovascular
diseases hence promoting good health (SDG 3) and ensuring no poverty (SDG 1).

4
 2 LITERATURE REVIEW

2.1 Passion fruit

2.1.1 Description, production and utilization

Passion fruit (Passiflora edulis) is a perennial fruit vine native to tropical America (Knight &
Winters, 1962). The ripe fruit is refreshing with delicate flavors, pleasing aromas and high nutritive
value. Passion fruit mainly exists in two forms; purple and yellow passion fruit.

Passion fruit has nutritive and medicinal benefits (Joy, 2010). It is a high acid food (pH = 3.2),
and a good source of nutrients; vitamins (A, B2 and C), phytochemicals, carotenoids, minerals (K,
P, Ca, Fe, Na, Cl, S) (Ramaiya, Bujang, Zakaria, & Saupi, 2018; Thokchom & Mandal, 2017;
Zibadi & Watson, 2004). Passion fruit plant contains anti-inflammatory, anticonvulsant,
antimicrobial, anticancer, anti-diabetic, antihypertensive, anti-sedative, antioxidant properties and
various remedial measures for treating conditions like osteoarthritis, asthma and act as colon
cleanser (Zas & John, 2016). It is, therefore, an attractive and nutritive fruit for both fresh
consumption and industrial processing.

According to SHEP PLUS ( 2016), the average yield of passion fruit in Kenya is about 4,000 kg
per acre per year which can be more than 6,000 to 8,000 Kg per acre with good management and
crop husbandry. According to IBGE, (2020), passion fruit has average yield of 14 tons ha-1 in
Brazil which varies among producers in different states and crop systems. The differences are even
greater when considering the profile of producers and also the productive potential of genetically
improved cultivars (Gontijo et al., 2019; Jesus et al., 2018; Cavichioli, Corrêa, Garcia, & Fischer,
2011). The yield reaches 60 tons ha-1 year-1 in open fields and 110 tons ha-1 year-1 in a greenhouse
(Gontijo et al., 2019; Jesus et al., 2018; Cavichioli et al., 2011). Minas Gerais state in 2018 showed
the fifth largest production of passion fruit in the country, with 31,500 tons in a harvested area of
2,135 ha (IBGE, 2020). This state has capacity to process up to 100 tons of passion fruit juice a
day (ABIR, 2020). According to (Conab, 2020), 1,270 tons of passion fruit were sold by Ceasa-
Uberlândia in 2018, with 75% of the fruits grown by producers in the Triângulo Mineiro. Besides
production, there is also increasing consumption of fruit juices, nectars and other fruit beverages
in Brazil, Europe and China.

5
Passion fruit can be consumed directly or processed into passion fruit-based products or blended
into other products. It is a highly acidic fruit (2.96-4.02 g of citric acid per 100g of juice) (Aline,
et al., 2011). It is thus, rarely used for direct consumption. It can be added to products like fruit
salads, ice cream, cool drinks, and concentrates. The fruit can be processed into juice (Lin, 2014;
Zhu, Duan, Yang, & Huang, 2017; Du, Li, & Chen, 2016), fruit wine (Pan, Qin, & Huang, 2011;
Nzabuheraheza & Nyiramugwera, 2014), jam (Kan & He, 2008), and fruit vinegar (Wang, Zhou,
& Chen, 2015; Pan, Huang, & Huang, 2012). Its peel can be processed into animal feeds or pectin
and dietary fibre (Zhu, et al., 2017). The wide application of passion fruit as a raw material for
several foods is due to its unique aroma (Zhu, et al., 2017). According to Zhu et al., (2017) passion
fruit juice processing is mainly based on the pulp and the nutrients of the pulp is added to the juice
giving the composition of the juice.

2.1.2 Passion fruit juice composition and processing

Passion fruit juice is a natural nutritious drink containing all the necessary essential nutrients
required by the body in the right proportion that meets the daily body requirements (Ramaiya,
Bujang, Zakaria, & Saupi, 2018). The juice contains carbohydrates, fibre, vitamins, minerals and
organic acids (Ramaiya et al., 2018; USDA, 2011). The trend in the composition of passion fruit
juice varies depending on the species and environment (Wardlaw, 2003). The proximate
composition of passion fruit juice from the different species are shown in the table 1. Passion fruit
juice has a moisture content of about 84-88 %, a good medium for enzyme action and general
metabolic processes (Ramaiya et al., 2018). The moisture is also important for stability and quality
of juices. Passion fruit contains good protein content and fiber compared to other fruits like
pineapple, oranges, papaya and apple (Ramaiya et al., 2018). Its protein content ranges between
0.67 % in P. edulis flavicarpa (USDA, 2011) and 2.81 % in P. edulis (Purple) (Ramaiya et al.,
2018). The juice also contains fiber (0.2 to 8.49±0.40 %) (Table 1). According to Chau and Huang
(2004), adequate comsumption of dietary fiber lowers the risk of coronary heart disease, diabetes,
colon and breast cancer, serum cholesterol level, constipation, hypertension. Passion fruit has low
fat content (<0.5%) just like most fruits (Ramaiya et al., 2018). Passion fruit juice also contains
carbohydrates which is lower in the mesocarp of P. quadrangularis (3.00±0.19 % ) compared to
juice of the different passion fruit species (e.g 14.25% in P. edulis flavicarpa and 6.57±0.29 in P.
quadrangularis) (Ramaiya et al., 2018; USDA 2011, NZ passion growers 2007). Passiflora fruit

6
juices with low carbohydrate are ideal for diabetic and hypertensive patients requiring low sugar
diets (Ramaiya et al., 2018).

Passion fruit juice is also rich in minerals. It has high ash content (0.34 - 1.8%) compared to other
fruits such as oranges, banana and papaya (Ramaiya et al., 2018). The high ash content indicates
the richness of passion fruit juice in minerals which is very important for proper functioning of the
body (Rao, 1996). Mineral composition of passion fruit juice from different species is shown in
the tables 2.

7
Table 1 Proximate composition of passion fruit juice (per 100 g)

Species Moisture content Ash Crude protein Crude fiber Crude fat Carbohydrate
(g) (g) (g) (g) (g) (g)
P. edulis (Purple) 85.45±0.56 1.18±0.11 2.81±0.19 4.31±0.31 0.50±0.14 6.95±0.73
P. edulis (Frederick) 84.37±0.63 1.03±0.03 2.40±0.11 3.47±0.26 0.51±0.10 7.44±0.69
P. maliformis 85.44±0.36 0.91±0.01 2.20±0.14 3.63±0.18 0.45±0.02 7.17±0.64
P. quadrangularis 86.16±0.46 1.37±0.14 2.35±0.09 2.40±0.11 0.35±0.0.01 6.57±0.29
P. quadrangularis (mesocarp) 86.63±0.33 0.51±0.02 1.13±0.11 8.49±0.40 0.24±0.01 3.00±0.19
P. edulis 84.7 1.00 2.80 3.30 0.50 7.39
P. edulis flavicarpa 84.21 0.70 0.67 0.20 0.18 14.25
P. edulis(wet basis) 87.5±0.014 0.34+0.014 0.23+0.014 ND ND 11.9+0.028
Sources: Ramaiya, Bujang, Zakaria, & Saupi (2018), NZ passion growers (2007), USDA (2011), Adeyeye, Ekiti, & Aremu (2017).

8
Table 2: Mineral composition (per 100g dry weight) of passion fruit juices

Species Ca Mg K Na P Cu Zn Fe
(mg) (mg) (mg) (mg) (mg) (mg) (mg) (mg)
P. edulis 54.91± 79.49 110.00±87.79 453.33±32.87 37.50±14.43 115.41±53.19 0.12±0.29 0.30±2.08 3.13±1.09
(purple)
P. edulis 51.75±72.17 108±52.04 402.50±42.56 20.83±44.09 117.92±48.05 0.11±0.89 0.28±1.53 2.98±0.93
(Frederick)
P. maliformis 54.33±58.33 170±76.38 362.5±25.25 39.17±50.69 237.50±36.08 0.11±2.08 0.46±0.88 1.22±0.33
P. 44.50±58.33 107.52±28.87 408.67±44.23 41.67±80.47 111.67±58.33 0.16±1.50 0.42±1.09 3.37±1.36
quadrangularis
P. 53.92±97.33 75.83±30.05 283.33±8.33 3.58±7.41 77.08±29.17 0.07±0.87 0.17±0.87 1.23±1.16
quadrangularis
(mesocarp)
P. edulis 12.00 29.00 348.00 28.00 68.00 0.09 0.10 1.60
(purple)
P. edulis f. 4.00 17.00 278.00 6.00 25.00 0.05 0.06 0.36
flavicarpa
P. edulis 16.00 39.00 350.00 28.00 54.00 Nd 0.10 2.42
P. edulis 10.4 0.73 24.7 22.6 26.6 0.12 0.8 1.10
Sources: Ramaiya et al., (2018), NZ passion growers (2007), USDA (2011), Adeyeye, Ekiti, & Aremu (2017).

9
The mineral composition of passion fruit juice varies with the species and environmental
conditions (Martin, 1997; Babasaheb, 2000; Wardlaw, 2003). Potassium is the most abundant
mineral in passion fruit just like in other fruits (Ramaiya et al., 2018). P. edulis (Purple) has the
highest amount of potassium (Table 2). This content is comparable with that of guava (417 mg/100
g) but higher than orange (181 mg/100 g) and pineapple (109 mg/100 g) (USDA 2011). Potassium
is important in controlling blood pressure and acid-base balance (Wardlaw, 2003). It is a co-factor
for many enzymes and is required for insulin secretion, creatine phosphorylation, carbohydrate
metabolism and protein synthesis (Ringer & Bartlett, 2007). P. maliformis has the highest amount
of phosphorus (237.50±36.08 mg/100 g) (Table 2). Phosphorus is important for metabolic
processes in the body and helps in strengthening the teeth and bones (Wardlaw, 2003). Phosphorus
together with calcium are important to the supportive structures of the body (Wardlaw, 2003;
Babasaheb, 2000). Magnesium is an important co-factor in enzyme systems and together with
calcium is important in neuro-chemical transmission and muscle excitability (Martin, 1997;
Wardlaw, 2003). Calcium is also important for bone formation and strength, and blood coagulation
(Ramaiya et al., 2018). Sodium in passion fruit juice is higher than in other fruits such as papaya
(8.00 mg/100 g) and guava (2.00 mg/100 g) (Ramaiya et al., 2018). A cup containing 274 g of
passion fruit juice supplies 3-6 % of the daily requirement of sodium for adult men and women
aged 19 to 30 years (Ramaiya et al., 2018).

Passion fruit is a rich source of non-heme or plant based iron (Ramaiya et al., 2018). Iron helps
during formation of red blood cells and is very important especially in the diet of infants and
pregnant women (Martin, 1997; Babasaheb, 2000; Oluyemi et al., 2006). A cup of P. edulis and
P. quadrangularis juices (1 cup containing 247g of passion fruit juice) provides more than 90%
for men and 40% for women of the recommended allowance for iron (Ramaiya et al., 2018). Most
studies did no detect managanese in passion fruit juice however, 0.09 and 0.22 mg of manganese
per 100g was reported by NZ Passion Fruit Growers (2007) and Adeyeye, Ekiti, & Aremu, (2017),
respectively.

Passion fruit juice also contains organic acids (Ramaiya et al., 2018). Organic acids play an
important role on the organoleptic properties, stability and microbiological quality of passion fruit
juice (Hasib et al., 2002). Passion fruit juice mainly contains citric, malic and acetic acids
(Ramaiya et al., 2018). According to Joy, (2010), passion fruit is also reported to contain lactic

10
acid:18.78 mg 100 g-1 in P. edulis (Purple) and 1.45 mg 100 g-1 in P. edulis f. flavicarpa. The level
of these acids varies depending on the species (Macrae, Robinson, & Sadler, 2007). Citric acid
which is the most abundant organic acid in passion fruit juice is higher in P. edulis (Purple)
(1487.30±0.28 mg 100 g-1) than in P. quadrangularis (1137.00±0.13 mg 100 g- 1) (Ramaiya et
al., 2018). Passion fruit juice has high citric acid (Fainsod, 2001) which together with other acids
makes it acidic. Citric acid is a weak, soluble organic acid that enhances flavor and increases
stability of soft drinks and syrups, and prevents color change by oxidation (Fainsod, 2001).

Passion fruit can be processed into pulp and blended beverages. According to Fernandes et al.,
(2011) at industrial level, passion fruit juice processing involves weighing, immersion of the fruits
into chlorinated water (25 mg mL-1) to disinfect the fruit and grading in terms of physical integrity,
color uniformity and degree of ripeness. The fruit is cut and pulp separated from the peel. A pulp
finisher is used to separate the pulp from the seeds. Formulation of the juice (addition of
preservatives such as sodium benzoate, sodium metabisulphite, potassium sorbate and acidulant
such as citric acid to the pulp) and homogenization is done. The juice is then deaerated using
vacuum deaerator at temperature of 50 °C and pressure of 600 mmHg. Pasteurization at 90 °C for
60 seconds in a tubular heat exchanger can then be done followed by hot filling at 85°C in bottles.
Capping and sealing are done and the juices are then stored.

2.1.3 Passion fruit juice blends

Passion fruit, whether pure or in combination with other juices, has good acceptability, and is
considered an important source of vitamins, minerals, soluble and insoluble fibers (Righetto,
Beleia, & Ferreira, 1999). Several studies have reported blending of passion fruit. Pruthi &
Girdhari., (1955) reported on the blending of passion fruit juice and squash. Nectar blends of
passion fruit and papaya was reported the best compared to those of mango and papaya pulp,
passion fruit and pear juice, mango pulp and pear juice, and pear juice and papaya pulp (Imungi &
Choge, 1996). The same study also reported blend of passion fruit and pear. Energy drinks from
mixed fruit nectar containing passion fruit (3.9%), papaya (5.7), guava (5.7) and acerola (5.7) were
acceptable (De Sousa, Maia, & De Azeredo, 2007). Salomon et al., (1999) reported high
acceptance of papaya-passion fruit mixed fruit nectar. Nectar from orange and passion fruit juices
is also reported which had a reduction in sensory acceptance of blends with higher proportion of
passion fruit due to the strong passion fruit flavor (Shaw & Wilson, 1988). Sousa et al., (2010)
11
produced a mixed fruit nectar blend composing of cashew apple, passion fruit, papaya, guava and
acerola fruit juices and caffein. This was acceptable, microbiologically stable with high vitamin C
content suggesting its potential for market. Much as blending of passion fruit juice with other foods
has been reported, there is scanty information on blending of passion fruit juice with flavourants
such as mint.

2.2 Mint

2.2.1 Description and production

Mint is a cultivated species in Europe and southwest Asia that belongs to the Lamiaceae family
(Moreno, Bello, & Primo-yu, 2002). The mint family is distinguished from others with a broad
square shaped stem (Elmastaş, Dermirtas, Isildak, & Aboul‐ Enein, 2006). Spearmint
(M. spicata L.) is a herbaceous, rhizomatous, perennial mentha species with flavor and pungent
taste because of the high concentration of essential oils such as carvone (Yadav, Kumar, & Dikshit,
2006). There are mainly three mentha species that have been cultivated around the world for food
and medicinal purposes. These are M. arvensis L. (cornmint), (M. × piperita L. (peppermint)
and M. spicata L. (spearmint) (Figure 1) (Moreno et al., 2002; Villasenor et al. 2002). Spearmint
(Mentha spicata L), also known as brown mint, garden mint, lady’s mint, and sage of Bethehem
is indigenous to northern England (Kuruppacharil, 2006). Spearmint is a creeping rhizomatous
(Jirovetz, Buchbauer, Shahabi, & MB, 2002), perennial herb with sharply serrated broad leaves
having a characteristic aromatic odour and pungent taste. Spearmint produces long pink or white
flowers in slender spikes. In contrast to Japanese mint and peppermint, Spearmint lacks the after-
cooling effect (Elmastaş, Dermirtas, Isildak, & Aboul‐ Enein, 2006).

Peppermint Corn mint Spearmint

Figure 1: Appearance of commercially important species of mint.

12
Spearmint is cultivated from climates ranging from tropical and temperate such as America, Brazil,
China, Europe and South Africa (Lawrence, 2007; Hajlaoui, Trabelsi, Noumi, & Snoussi, 2009).
Spearmint is widely grown in the entire regions worldwide (Lawrence, 2007). Peppermint and
spearmint are the main mint crops produced in US (Vining et al., 2020). According to Food and
Agriculture Organization of the United Nations (FAOSTAT, 2018), Morocco, Argentina, Mexico,
Bulgaria, and Spain have reported largest peppermint production. There is no data available on
mint production in Uganda. In 2018, world mint oil production was 106,728 MT (FAOSTAT,
2018). Annual farm gate value of peppermint and spearmint production in the US ranged from
$100 to $150 million and $45 to $50 million, respectively during the past decade (NASS, 2020).
Highest US mint oil production was in 2011, at about 7 million pounds (3,175 MT) of peppermint
oil and 2.2 million pounds (998 MT) of spearmint oil (NASS, 2020). Since then, US production
has dropped to under 6 million pounds (2,721 MT), as production of menthol from oils of M.
arvensis in China and India has been increasing. The expansion of market has been driven by the
demand for mint oil by Southeast Asian countries (Liu and Lawrence, 2006).

2.2.2 Utilization of mint

Mint has a wide range of applications in the medical, food, pharmaceutical world. It has been used
as a medical and aromatic plant by eastern and western cultures since ancient times (Park,
Vohnikova, & Brod, 2002). Spearmint is used broadly in cosmetics and soaps (Bensabah,
Houbairi, Essahli, Lamiri, & Naja, 2013), toothpaste, breath fresheners and antiseptic mouth rinses
(Kumar, Mishra, Malik, & Satya, 2011; Tyagi & Malik, 2011). It is also used as an antimicrobial
(Sulieman, Abdelrahman, & Rahim, 2011), antioxidant (Mata et al., 2006), and insecticide
(Samarth & Kumar, 2003). In the medical world, spearmint is used as a herbal medicine for treating
respiratory tract diseases, colds and flu, gastralgia, hemorrhoids, and stomachache (Asekun,
Grierson, Afolayan 2007; Kanatt et al., 2007). According to Bensabah, Houbairi, Essahli, Lamiri,
& Naja, (2013), carminative properties of spearmint taken as tea is important for digestive
disorders, fever (Kuruppacharil, 2006), nausea, vomiting and gastrointestinal disorders (Kumar,
Mishra, Malik, & Satya, 201;1 Tyagi & Malik, 2011).

Mint has also been widely used in food, confectionary and chewing gum industries (Igoumenidis,
Lekka, & Karathanos, 2016; Bensabah, Houbairi, Essahli, Lamiri, & Naja, 2013). It is used to
preserve (Scherer, Lemos, Lemos, Martinelli, Damasceno, et al., 2013), impart aroma and taste
13
(Igoumenidis, Lekka, & Karathanos, 2016). It has been used to flavor foods products like cheese
and dough (Singh, Shushni, & Belkheir, 2015). It has also been utilized as an ingredient in herbal
teas (Lv, et al., 2012), ash gourd Benincasa hispida) juice (Majumdar et al., 2008), grapes juice
(Poornimah & Ritu, 2018) and pineapple juice and guava leaves extract (Shalini & Ritu, 2008).
Mint has also been used in Indian and Italian cuisine in dried and fresh form to fish and shellfish
plates before or after cooking (Kizil, Hasimi, Tolan, Kilinc, & U, 2010). There is currently no
literature on use of mint in passion fruit juice.

2.2.3 Composition and use of mint oil and hydrosol

Mint essential oil (EO) contains several compounds (table 3). Mentha piperita (peppermint),
Mentha spicata (native spearmint) and Mentha gracilis (Scotch spearmint) EO mostly contain
monoterpenes and their derivatives (76–90%), but differ in major compounds (menthol and
menthone (50%) in peppermint and carvone (70%) in spearmint EO) (Tortajada, Cao, & Ji, 2019).

14
Table 3 Compounds found in mint essential oil
Mint type Major compounds References
Spearmint essential oil Carvone (40.8 – 76.65%), limonene (1.59% -20.8%), pulegone (Dhifi, Jelali, Mnif, Litaiem, & Hamdi,
(24.72% to 26.67%), mentone (21.92%), piperitenone oxide (12.55 - 2020; Bardaweel et al., 2018; Franciscato et
25.84%), muurolene (2.29 - 2.3%), myrcene (0.59 - 2.1%), beta- al., 2018; Adjou, Gbaguidi, Dahouenon-
caryophyllene (1.2% - 2.7%), 1,8-cin- eole (3.6-17%), β-pinene (0.69- ahoussi, & Soumanou, 2017; Sevindik,
2.2%), cis dihydrocarvone (0.2-3.9%), germacrene D (0.9-2.1%), Yamaner, Kurtoğlu, & Tin, 2017; Snoussi
linalool (0.48-58.51%), 4-terpineol (1.3% - 5.73 %), beta-bourbonene et al., 2015; Scherer, Lemos, Lemos,
(1.18%), trans-dihidrocarvone (1.03%), iso- dihydrocarveol (3.4%), Martinelli, Martins, et al., 2013; Şarer,
cis-Calamenene (0.72%), L-menthone, (32.74%), menthol (11.42%), Toprak, Otlu, & Durmaz, 2011; Chauhan,
borneol (0.1 -7.17%), alpha-cadinol (0.21 - 1.52%) dihydrocarveol Nautiyal, & Tava, 2010; Sokovic & Van
(0.06 – 3.29%), sabinene (0.14 - 1.4%), piperitone (0.9 – 6.08%), cis- Griensven, 2006; Kokkini, Karousou, &
Isopulegone (1.69) Lanaras, 1995)
Mentha longifolia L Trans-dihydrocarvone (23.64%), piperitone (17.33%) and cis- (Džamić et al., 2010)
essential oil dihydrocarvone (15.68%)
Peppermint leaves oil 50-78 % menthol, monoterpene, Menthofurane and traces of jasmine (Dew & Evans, 1984)
Mentha piperita hydrosol Menthols and menthones (Garneau, Collin, & Gagnon, 2014)
Peppermint essential oil Dominated by α-Phellandrene acid and β-Bourbonene, followed by (Siddeeg, Salih, & Mukhtar, 2018)
Trans-carveol, Methyl caproate (C6) (Fresh sample) and α-Humulene,
Bornylacetate followed by Cis-Limonene oxide, 4, Methyloleate (C
18:1) (Dry sample)
Mentha piperita Menthol (Singh, Shushni, & Belkheir, 2011)
Essential oil of dried aerial Piperitenone (29.9%), pulegone (13.9%) and 1, 8-cineole (10.5%) (Wojcik-Stopczynska, Jakowienko,
parts of (Mentha crispa L.) Wysocka, 2012)

15
Mints have variations in their essential oil chemical composition (table 3). The main component
reported is carvone in M. spicata, and menthol in M. piperita. Concentration of these components
also vary with geographical locations (Viljoen, Petkar, VanVuuren, Figueiredo, Pedro, Barroso,
2006). Natural variation in the composition of essential oil, many factors in growing conditions,
such as temperature, climate, humidity, radiation, soil type season and the time of collection of the
plant material may also lead to these variations (Viuda-Martos, Ruiz-Navajas, Fernández-López,
& Perez-Álvarez, 2008; Shanjani, Mirza, Calagari, & Adams, 2010; Butkiene & Mockute, 2011).

Carvone, which is a phenolic compound, is the main compound found in the essential oil of
spearmint, followed by limonene (Snoussi et al., 2015 ; Jirovetz, Buchbauer, Buchbauer, Shahabi,
& MB, 2002). Carvone helps in inhibiting bacterial growth (Helander, et al., 1998), acts as insect
repellent (Lee, Tsao, Peterson, & Coats, 1997) and fungicide (Smid, de Witte, & Gorris, 1995).
The odor of spearmint and caraway seeds is due to (S)-carvone and (R)- carvone, respectively
which are enantiomers of carvone with different response. S-carvone also has high antioxidant
activity (Elmastaş, Dermirtas, Isildak, & Aboul‐ Enein, 2006).

2.2.4 Antioxidant properties and other health benefits of mint

Mint has antioxidant properties (Džamić et al., 2010). This is mainly reported in the oil (Snoussi
et al., 2015), and methanolic extracts of the leaves (Scherer, Lemos, Martinelli, Martins, et al.,
2013; Al-juhaimi & Ghafoor, 2011; Kanatt et al., 2007). The leaves and stems of mint have high
quantities of total phenols (>1.02 mg GAE/100 ml) and antioxidant capacity. The antioxidant
activity of mint extract is higher than that of synthetic Alternative oxidase (AOX) and is quite
similar to Butylated hydroxyl anisole (BHA). The high antioxidant capacity is mainly due to the
presence of high amounts of vitamin E, total phenolic compounds, flavonoids and fatty acid methyl
esters in mint (Rameshwar, Ismail, Yeng, Sasidharan, & Kumar, 2012).

Spearmint oil has been used in medicine due to its several other health benefits. It has been used
as a herbal medicine in folkloric remedies for treating colds and flu, respiratory tract problems,
hemorrhoids, gastralgia and stomach ache (Tetik, Civelek, & Cakilcioglu, 2013; Asekun, DS, &
Afolayan, 2007; Kanatt, Chander, & Sharma, 2007). According to Bensabah, Houbairi, Essahli,
Lamiri, & Naja, (2013), spearmint tea has carminative properties which help to treat digestive
disorders, fever and minor ailments (Kuruppacharil, 2006). Spearmint has also been reported to
treat nausea, vomiting and gastrointestinal disorders (Kumar, Mishra, Malik, & Satya, 2011; Tyagi

16
& Malik, 2011). It improves memory (Adsersen, Gauguin, Gudiksen, & Jager, 2006), and is a
stimulant (Papachristos & Stamopoulos, 2002). Spearmint also has several biological uses. It has
been used as insecticides (Samarth & Kumar, 2003), antimicrobials (Ozgen, et al., 2006),
antioxidants (Choudhury, Kumar, & Garg, 2006), antispasmodics, and anti-platelets (Tognolini, et
al., 2006). These several health benefits make spearmint to be widely used in foods or directly
consumed as medicine.

2.2.5 Antimicrobial properties and other effects of mint on composition, and stability of
foods
Mint has antimicrobial effects against several microorganisms (Franciscato et al., 2018; Snoussi
et al., 2015; Şarer, Toprak, Otlu, & Durmaz, 2011; Scherer, Lemos, Martinelli, Martins, et al.,
2013; Sokovic & Van Griensven, 2006). Most of the mint species possess antimicrobial activity
as shown in table 4. According to Kadoglidou, et al. (2011), the antimicrobial activity of mint is
associated with the presence of terpenoids. Spearmint has terpenes and terpenoids as the main
chemical groups. Several studies have reported that terpenes do not represent a group of
constituents with high inherent antimicrobial activity. For example, Koutsoudaki, Krsek, &
Rodger, (2005) reported low or no antimicrobial activity of ß-pinene, p-cymene, ß- myrcene,
limonene, and γ-terpinene against Escherichia coli, Staphylococcus aureus, and Bacillus cereus.
p-cymene and γ-terpinene were also reported ineffective as fungicides against Saccharomyces
cerevisiae (Rao, Zhang, Muend, & Rao, 2010). This is contradicting with several studies which
reported terpenoids as a group of antimicrobial compounds, active against wide range of
microorganisms (Dorman & Dean, 2000). This is linked to their functional groups. The hydroxyl
group and the delocalized electrons are vital for the antimicrobial activity (Dorman & Dean, 2000).
According to Bassolé, et al., (2010) menthol, thymol and linalool possess antimicrobial activity
against Listeria monocytogenes, Enterobacter aerogenes, Escherichia coli, and Pseudomonas
aeruginosa. This antimicrobial activity of essential oils can be very important for prolonging shelf
life of foods.

17
Table 4 Antimicrobial properties of mint
Mint variety/sample Microbes acted upon References
Pepper mint EO Staphylococcus aureus and Salmonella E. coli and Klebsilla pneumonia (Siddeeg et al., 2018).
Spearmint oil Staphylococcus epidermidis (Bardaweel et al., 2018).
Spearmint oil from Aspergillus parasiticus and Fusarium versicolor; and Mucor spp. (Adjou et al., 2017)
Benin
volatile vapours of Staphylococcus aureus, Escherichia coli, Bacillus subtilis, and Pasturella multocida, (Yadav et al., 2006).
Mentha spicata Aspergillus niger, Mucor mucedo, Fusarium solani, Botryodiplodia theobromae, and
L.var. MSS-5 Rhizopus solani
essential oil
M. pulegium and M. Methicillin-resistant Staphylococcus aureus, Staphylococcus aureus ATCC 6538, (Sevindik et al., 2017)
spicata subsp. Pseudomonas aeruginosa, Enterococcus faecium DSM 13590, Escherichia coli O157:H7
Tomentosa and Bacillus cereus CCM99
Peppermint oil Gram-negative and Gram-positive bacteria (Singh, Muftah, & Shushni, 2011). (Sartoratto, Machado, &
Salmonella spp. Delarmelina, 2004).

Mentha longifolia Aspergillus and Fusarium species, Penicillium funiculosum, Trichoderma viride, (Džamić et al., 2010).
(L.) Hudson Trichophyton menthagrophytes, Candida albicans, Alternaria alternate, (Cladosporium
(Lamiaceae) EO fulvum, Cladosporium cladosporioides and Penicillium ochrochloron).
Essential oil of dried Highest inhibitory action against Botrytis cinerea and Cladosporium herbarum, and the Wojcik-Stopczynska,
aerial parts of least–against Aspergillus parasiticus and Fusarium oxysporum. Higher amounts inhibited Jakowienko, Wysocka, (2012)
wrinkled-leaf mint the growth of Aspergillus fumigatus
(Mentha crispa L.)

18
Majumdar et al., (2008) reported microbial stability of ash gourd (Benincasa hispida) and mint
leaves (Mentha spicata) juice (in ratio 75:25) during 6 months of storage at room temperature
(28±2 °C). Mentha spicata L essential oil exhibited high antifungal activity against Aspergillus
parasiticus, Fusarium versicolor and Mucor spp. in mango puree stored at 4°C and 25°C (Adjou
et al., 2017).

Besides antimicrobial effect of mint on foods, other effects on the property of foods has also been
reported. Kanatt et al., (2005) reported significant retardation (p < 0.05) of lipid oxidation in meat
stored at chilled temperatures for 4 weeks. Pineapple juice and guava leaves extract flavored with
mint was reported the best in terms of carbohydrate, energy, calcium and iron content compared
to the same beverage flavored with dates (Shalini & Ritu, 2008). Mint used to flavor grape juice
also increased antioxidant, iron, carotene, calcium and vitamin C content of herbal mint flavored
grapes juice (Poornimah & Ritu, 2018). These effects of mint could result into improvement on
the composition and stability of passion fruit passion fruit juice.

19
 3 MATERIALS AND METHODS

3.1 Sample preparation

3.1.1 Raw materials

Fresh fully ripened purple passion fruits (Passiflora edulis), and tender fresh spearmint (Mentha
spicata L.) leaves were obtained from Gayaza market (Uganda) and brought to the School of Food
Technology, Nutrition and Bio-engineering, Makerere University.

3.1.2 Preparation of the mint extract

Mint extract was prepared from fresh mint leaves. The leaves were removed from the stalks,
washed using potable water, weighed (1.3 Kg), blanched for 2 minutes and blended with equal
amount of fresh warm water (40 °C). The extract was sieved using a clean muslin cloth and stored
at -20 °C for 12 h.

3.1.3 Preparation of mint hydrosol

Mint hydrosol was prepared using steam distillation process. Steam was passed through 15 kg of
the aerial parts of spearmint plant. Volatile essential oil and steam were condensed and the oil
which settled on top of the water separated. The spearmint hydrosol was then collected. Five litres
of hydrosol and 15 mL of essential oil were obtained from steam distillation of spearmint.

3.1.4 Preparation of the passion fruit juice

Ripe purple passion fruits were weighed, washed in potable water containing 0.05% sodium
hypochlorite. The fruits were cut into two halves using a knife and the pulp together with the seeds
scooped out using a spoon. The pulp was obtained by blending (using a blender) with water in the
ratio 1:3 (pulp: water). It was then filtered using a clean muslin cloth.

20
3.1.5 Preparation of mint flavored passion fruit juice

Sugar (sucrose) (about 378 g) required to obtain 12°Brix, 0.2 % Carboxy Methyl Cellulose
(Shanghai, China), 0.1 % citric acid (Shanghai, China) and 0.03 % potassium sorbate (Shanghai,
China) were added to the juice (section 3.1.4). The juice was then blended with mint extract or
hydrosol in the ratios shown in Table 5. The juices were pasteurized at 90 °C for 1 min and hot
filled in 300 mL plastic bottles previously disinfected with 0.05% sodium hypochlorite. They were
then stored at room temperature (25 °C) for 140 days.

Table 5 Formulations of mint flavored passion fruit juice

Treatment Passion fruit juice : mint Treatment Passion fruit juice : mint hydrosol
extract
T0 100 : 0 100 : 0
T1 96 : 4 T5 96:4
T2 92 : 8 T6 92 : 8
T3 88 : 12 T7 88 : 12
T4 84 : 16 T8 84 : 16

3.2 Determination of effect of spearmint extract and hydrosol concentration on sensory

acceptability of the prepared juices

The mint flavored passion fruit juices were subjected to sensory acceptability tests by 25 semi-
trained panelists chosen from among Makerere University students. A nine-point hedonic scale
(1= dislike extremely, 2= dislike very much, 3=dislike moderately, 4= dislike slightly, 5 neither
like nor dislike 6= like slightly, 7= like moderately, 8=like very much, 9= like extremely) was used
by the panelists. Panelists rated the acceptability of the different attributes of the juices such as
appearance, color, aroma, taste and overall acceptability using a provided questionnaire (appendix
VII). Panelists were provided with mineral water to rinse the pallet before and in between tasting
of samples. The most accepted passion fruit juices from the sensory evaluation were selected for
nutritional composition and shelf-life studies.

21
3.3 Determination of effect of spearmint extract and hydrosol concentration on the
nutritional composition and antioxidant property of passion fruit juice

3.3.1 Nutritional Composition

Vitamin C

Vitamin C content of the samples was determined using Dichlorophenolindophenol (DCPIP)

titrimetric method (AOAC 967.21, 45.1.14, 2007). Five milliliters of the sample were pipetted into
a 50 mL volumetric flask, topped to the mark using 5% extracting solvent and then shaken
vigorously. Five milliliters of the extract were titrated against DCPIP solution until the solution
turned pink. The vitamin C content of the sample was calculated using the formula

Titre Value × 0.0651 × total Volume

Vitamin C content (mg/100ml) = × 100
sample volume × Volume pipetted for titration

Beta-Carotene

The beta-carotene content of the juice was determined using the Harvest plus a method as
described by Rodrigues-Amaya & Kimura (2004). Five milliliters of the juice were pipetted into a
50 mL volumetric flask. Fifty milliliters of acetone were used to extract the carotenoids using
motor and pestle in acetone. The extract was filtered in volumetric flask through a glass wool in a
funnel until 50 ml mark. Thirty milliliters of Petroleum ether (PE) were added to the separating
funnel containing 50 mL of extract. About 900 mL of distilled water were added into separating
funnel at intervals of 200 mL and water separated from extract by opening the tap. The PE-
carotenoid extract was run through a funnel containing cotton wool and anhydrous sodium sulphate
to absorb any remains of water in the extract. Remains of the extract in the separating funnel were
rinsed using PE into the flask until 50 mL mark. The extract was shaken, covered and kept in dark.
Absorbance of the extract was measured using spectrophotometer at 450 nm. Beta-carotene
content of the juices was calculated using the formula below;

Absorbance × Total Volume × 10,000

Beta Carotene (µg/100mL) = × 100
2569 × Sample Volume

22
3.3.2 Anti-oxidant property

Total anti-oxidant in the juice was determined using DPPH method (AOAC, 2012.04). Five
millilitres of extracting solvent (80% acidified methanol) was added to 1 mL of the juice and
incubated at 50 °C for 30 min. This was then frozen for 10 min and centrifuged (Thermo fisher
scientific, Heraeus megafuge 8, China) at relative centrifugal force of 3260×g (4500 RPM) for 10
min. The supernatant was decanted and extraction repeated. The pool was mixed with the
supernatant and centrifuged at a force of 3260×g for 5 min. Color was then developed by adding
0. 9 mL of acidified methanol and 2.9 mL of DPPH and absorbance read at 517 nm. Different
volumes (0.05-1 mL) of 0.01g per 100 mL of ascorbic acid were pipetted and 80% acidified
methanol (0.95-0 mL) added to make 1 mL. DPPH (2.9 mL) was added and then incubated for 30
minutes. Absorbance was measured at 517 nm. Standard curve of equation of line 𝑦 = 30.023𝑥 +
0.2862 was used to obtain the slope and intercept. Total antioxidant was determined using the
formula below;

Total antioxidants (mg ascorbic acid equivalent /100mL)

Actual absorbance − intercept × dilution factor × total volume
= × 100
sample volume × slope

Where actual absorbance = DPPH absorbance at 517nm −

(absorbance of sample at 517nm + absorbance of 80% acidified methanol)

3.4 Determination of effect of spearmint extract and hydrosol concentration on storage

stability of passion fruit juice

Juices formulations containing 0% mint, 4% mint extract, 8% mint extract, 4% mint hydrosol, 8%
mint hydrosol which were the most acceptable were selected for shelf stability studies. Juices were
then stored at room temperature (25 °C) for 140 days. Physicochemical analyses were done at
intervals of 0, 35, 75, and 140 days whereas microbial analyses were done at 0, 35, and 70 days of
storage.

23
3.4.1 Physicochemical analyses

pH

The pH of the juice was measured using a pH meter (ISFET pH meter, model IQ125 mini LAB,
Japan) calibrated with buffered solutions (pH 4.0 and 7.0) according to AOAC official Method
981.12, (1981). Two drops of the sample were added on to the probe of calibrated pH meter and
reading on the display screen recorded.

Titratable acidity (TA)

Titratable acidity was determined as described by the AOAC official method 942.15, (2000). Three
drops of phenolphthalein were added into 5 mL of the juice in a conical flask and titrated against
0.1N NaOH. The volume of NaOH used was noted and the acidity of the sample was calculated
as % citric acid using 0.064 as acid conversion factor.

volume of NaOH × Normality of NaOH × Acid Conversion Value

TA = × 100
sample volume

3.4.2 Microbial storage stability

Total Plate Count (TPC)

The total plate count was determined according to the International Standard -ISO 4833, 2003
method. Plate Count Agar (Laboratorios CONDA, Madrid, Spain) was dissolved in distilled water,
autoclaved at 121 C° for 15 min and then cooled to 45 - 47°C in a water bath. Serial dilutions were
carried out on the juice using quarter strength ringers lactate diluent, (Laboratorios CONDA,
Madrid, Spain). One millilitre of inoculum was aseptically transferred to the center of each petri
dish. About 20 mL of molten agar was poured into each petri dish and the inoculum carefully
mixed with agar by rotating. The mixture was allowed to solidify, petri dishes inverted and then
incubated at 37 °C for 24 h. Using a colony counter, counts ranging from 30-300 were taken and
results expressed as cfu/mL.

24
Yeasts and Molds

Yeast and molds were determined using surface spread technique (International Standard-ISO
21527-2, 2009). Potato dextrose agar (Laboratorios CONDA, Madrid, Spain) was thoroughly
mixed with distilled water and sterilized by autoclaving at 121°C for 15min. The medium was
immediately cooled in a water bath at 45 - 47 °C. One percent sterile lactic acid was added to the
sterile molten PDA. About 20 mL of molten agar was aseptically poured on to the petri dishes and
left to set at room temperature. The plates were inverted and serial dilutions prepared. Using a sterile
pipette, 0.1 mL of inoculum was transferred onto the center of solidified PDA and inoculum spread
using a sterile spreader. The petri dishes were incubated upright at 30 °C for 3-5 days. Using a colony
counter, counts ranging from 30-300 were taken and results expressed as cfu/mL.

Total coliforms

Total coliforms were determined using colony count technique (International Standard-ISO 4832,
2006). The Violet Red Bile Lactose (VRBL agar, Laboratorios CONDA, Madrid, Spain) was
thoroughly mixed with distilled water, heated until boiling with occasional stirring and allowed to
boil for 2 min. The molten VRBL media was cooled to 45 – 47 °C in a water bath. Serial dilutions
were carried out using quarter strength ringers lactate diluent, (Laboratorios CONDA, Madrid,
Spain). About 20 mL of molten agar was aseptically poured on to the petri dishes and left to set at
room temperature. One milliliter of inoculum was added to the center of the petri dish and about
10 mL of molten Violet Red Bile Lactose (VRBL agar, Laboratorios CONDA, Madrid, Spain) into
the petri dishes. The inoculum was carefully mixed with the agar and allowed to solidify. Control
plate was prepared with about 20 ml of the agar. Dishes were inverted and incubated at 30 °C for
24 h. Counts of purplish colonies were taken using a colony counter and results expressed as cfu/
mL.

3.5 Data analysis

All the experiments were done in duplicate. Microsoft Windows Excel 2019 and XLSTAT 2020
software was used for analysis of sensory, nutritional composition and shelf-life study data. One-
way Analysis of Variance (ANOVA) was used to check significant differences among designated
treatments (p < 0.05).

25
 4.0 RESULTS AND DISCUSSION

4.1 Effect of spearmint extract and hydrosol concentrations on the sensory acceptability
of passion fruit juices

Generally, sensory acceptability of the juices reduced with increase in concentration of mint
extract or hydrosol (Figure 2). The control, and juices containing 4% and 8% mint were the most
acceptable.

Figure 2: Effect of mint extract (a) and hydrosol (b) on the sensory acceptability of the passion
fruit juice

There was no difference (p>0.05) between scores for aroma and taste of passion fruit juices
flavored with mint extract. Appearance and color scores of the juices flavored with mint extract
were lower (p<0.05) than that of the control. Juice containing 4% mint extract was the most
acceptable (p < 0.05) among mint extract flavored juices. According to Bajaj, Urooj, &
Prabhasankar, (2006), color measurement of mint powder and mint extract-biscuits were lower
than mint menthol biscuits and control due to green color of mint. The change in appearance and
color (green) could have resulted into lower acceptability of the passion fruit juice flavored with
mint extract since passion fruit juice is typically associated with a yellow-orange colour. Mint
extract introduced chlorophyll which changed the color of passion fruit juice to a more undesirable
hue. This implies that lower concentrations of mint extract should be used in passion fruit or

26
chlorophyll should be removed in order not to affect the color and/or overall acceptability of the
juice. Singh & Duber, (2018) reported acceptability of grape and celery juice flavored with ≤ 8%
mint extract and juice with 8% mint extract was most acceptable followed by 6%, and 4% mint
extract. Acceptability of guava fruit juice and whey based beverages flavored with ≤ 6% mint
extract was also reported and 6% mint extract was the most acceptable (Maya et al., 2016). Maurya
& Ritu, (2018) reported acceptability of pineapple-guava leave extract beverage containing upto
10% mint extract. The beverage containing 6% mint extract was the most acceptable. These studies
reported mint extract concentrations of 8% or less as the most acceptable in foods quite similar to
the one in this study.

There was no difference (p>0.05) between the acceptability scores for appearance, color, and
aroma of mint hydrosol flavored passion fruit juices and control. Passion fruit juices containing 12
% or less mint hydrosol were liked (score of >5) with regards to taste. The taste of passion fruit
juice containing 16% mint hydrosol was disliked (score of <5). This may be due to 1-menthol in
the spearmint hydrosol which could have, in higher concentrations affected the acceptability of the
passion fruit juice. 1-menthol in spearmint oil causes bitterness (EP 0 690 678 B1, 1997).
According to European Patent Specification (EP 0 690 678 B1, 1997), removal of at least 25% of
1-menthol content of mint flavor reduced bitterness of mint flavor in chewing gum. 1-menthol in
mint flavor can be removed by distilling mint oil into top, middle and bottom fractions and then
removing portion of the middle fraction containing 1-menthol (EP 0 690 678 B1, 1997).
Acceptability scores for taste of juice containing 4% mint hydrosol was not significantly different
from the control (p>0.05). Juices containing 8%, 12% and 16% mint hydrosol scored significantly
lower than the control. All the mint hydrosol flavored juices were accepted by the consumers (score
of > 5) except that containing 16% mint hydrosol. Acceptability score of juice containing 4% mint
hydrosol was not significantly different from the unflavored juice (p>0.05). Acceptability of
samples containing 8%, 12% and 16% mint hydrosol was significantly lower than unflavored
passion fruit juice (p<0.05). However, there were no reports in literature to compare acceptability
of mint hydrosol in passion fruit juice and other foods. The concentrations of less than or equal to
8 % mint extract or hydrosol is, therefore, recommended to produce an acceptable mint flavoured
passion fruit juice. Basing on sensory results, juices that were selected for nutritional composition
and shelf study analysis were 0% mint extract, 4% mint extract, 4% mint hydrosol, 8% mint extract
and 8% mint hydrosol (appendix I).

27
 4.2 Effect of spearmint extract and hydrosol concentration on the nutritional composition
and antioxidant property of passion fruit juice

Vitamin C content of juices (Figure 3) ranged between 3.26±0.65 and 7.81±0.00 mg/100 mL.
Vitamin C content of passion fruit juice significantly increased with increase in mint extract
concentration. The Vitamin C content of the juices containing hydrosol was not significantly
different from the control.

9
Vitamin C Content (mg/100ml

8
7
6
5
4
3
of juice)

2
0% mint 4%mint 4% mint 8% mint 8% mint
Extract hydrosol extract hydosol
Mint concentration in the juice (%)

Figure 3: Vitamin C content of the mint flavored juices.

Several studies reported increase in vitamin C content of foods incorporated with mint. According
to Singh & Duber, (2018), vitamin C content of grape and celery juice flavored with 10% herbal
mint extract increased from 6 mg to 28.55 mg/100 mL. Vitamin C content of a pineapple based
beverage incorporated with gauava leave extract and mint extract ranged from 29.89 to 31 mg/100
mL (Maurya & Ritu, 2018). Maya et al., (2016) reported higher ascorbic acid content of guava
fruit juice and whey based beverage flavoured with mint (13.32 mg/100mL) than the unflavored
beverage (11.42 mg of ascorbic acid/100mL). The increase in vitamin C content of the juice is
attributed to addition of spearmint that had an additive effect on the vitamin C content of the juice.
The difference in vitamin C content of passion fruit juice containing mint extract and hydrosol

28
could be due to higher concentration of vitamin C in the mint extract than the hydrosol. The
hydrosol is not expected to contain vitamin C since it is prepared by distillation processes of mint
extracts. This implies that mint extract could be used to improve the vitamin C content of passion
fruit juice and other related juices.

Beta-Carotene content of the samples ranged between 45.98±3.11 and 104.81±0.43 µg/100 mL of
juice (Figure 4). Beta-Carotene content of passion fruit juice significantly increased with increase
in mint extract concentration in the juices (Figure 4).

120
B-carotene (µg/100 mL of juice)

110
100
90
80
70
60
50
40
30
20
0% mint 4%mint 4% mint 8% mint 8% mint
Extract hydrosol extract hydosol
Mint concentrations (%)

Figure 4: Effect of Mint on beta-carotene content of passion fruit juice

A similar trend was observed in a study by Singh & Duber, (2018) who reported an increase in
carotene content of the guava and celery juice from 2 to 244.4 µg when flavored with up to 10 %
mint extract. There was no significant change in the beta-carotene content of the juice by adding
4 % mint hydrosol however, addition of 8 % mint hydrosol significantly decreased beta-carotene
concentration of passion fruit juice. This could be due to dilution of the juice by hydrosol since it
only contains water and essential oil.

29
Total antioxidant level of the juices ranged between 300.70±0.00 and 303.04±0.34 mg ascorbic
acid equivalent (AAE)/100mL of juice (Figure 5). Generally, there was a significant (p<0.05)
increase in the total antioxidant level of the juice as concentration of the mint extract and hydrosol
in the juice increased. Addition of 4% mint hydrosol did not significantly increase the total
antioxidants in the juice (p>0.05).

304
Total antioxidants (mg AAE/100ml)

303.5
303
302.5
302
301.5
301
300.5
300
0% mint 4%mint 4% mint 8% mint 8% mint
Extract hydrosol extract hydosol
Concentration of mint

Figure 5: Effect of mint extract and hydrosol on total antioxidant content (Ascorbic Acid
equivalent AAE/100ml) of passion fruit juice.

Increase in antioxidants of foods incorporated with mint has been reported in many studies but
there is scanty information on effect of mint on total antioxidants of passion fruit juice. Maurya &
Ritu, (2018) reported a significant increase in total polyphenol and flavonoids antioxidants (131.47
– 225.48mg/100mL, and 147 – 293.58 mg/100mL respectively) in pineapple juice and guava
leaves extract flavored with up to 10% mint extract. Singh & Duber, (2018) reported an increase
in vitamin C and carotene antioxidants from 2 mg/100 mL and 6 µg/100 mL to 28.14 mg/ 100 mL
and 244.5 µg/100 mL, respectively as amount of mint increased in grape juice. Spearmint extract
exhibited excellent antioxidant activity in radiation processed lamb meat (Kanatt et al., 2005).
Application of mint hydrosol increased the total phenolics and antioxidants of carrot (Panayiota,
30
Antonios, George, & Nikos, 2018). Antioxidant activities of essential oils from aromatic plants
are mainly attributed to the active compounds present in them. This can be due to the high
percentage of main constituents, presence of other constituents in small quantities or synergy
among them (Siddeeg et al., 2018). The antioxidant efficacy of aqueous extract from Mentha
species is strongly associated with the phenol content (Dorman, Kosar, Kahlos, Holm, & Hiltunen,
2003) though individual antioxidants in spearmint for example flavonoids, vitamin C, carotenoids
also contribute significantly to the overall antioxidants in the product.

4.3 Effect of spearmint extract and hydrosol concentration on storage stability of passion
fruit juice

4.3.1 Physicochemical storage stability

The pH of the juice samples reduced from 3.1 to 2.65 during storage (Figure 6a). Generally, pH of
passion fruit juice was not significantly (p>0.05) affected by addition of up to 8% mint extract and
hydrosol though there was a slight reduction in the pH of all the samples during 140 days of
storage. Generally, the titratable acidity of the juices increased from 0.79±0.01 to 1.31±0.02 g/100
ml during storage (Figure 6b).

Figure 6: Effect of mint extract and hydrosol on the pH (a) and titratable acidity (b) of passion
fruit juices during storage at 25°C for 140 days

31
Majumdar et al., (2008) reported a reduction in pH of ash gourd-mint leaves juice from 4.0 to 3.93
during 6 months storage at room temperature (28±2 °C). The same study reported a significant
increase in total acidity of ash gourd-mint leaves from 0.25 to 0.32 g/100 mL during storage at
room temperature (28±2 °C). However, these slight changes in pH and titratable acidity did not
affect the product quality (Majumdar et al., 2008). Addition of mint extract and hydrosol did not,
therefore, affect storage stability of passion fruit juice with respect to pH and titratable acidity.

4.3.2 Microbial stability

The juices remained microbiologically stable during 70 days of storage. No coliforms (appendix
IV) and yeasts and molds (appendix III) were detected in the samples during 70 days of storage.
All TPC was <20 cfu/ml (appendix II) during 70 days of storage. Maya et al., (2016) reported < 4
colonies of bacteria, yeast and molds per ml of guava fruit-whey based beverage flavored with
mint. Majumdar et al., (2008) also reported low TPC (<10 cfu/ml) and no counts of yeasts and
molds in ash gourd–mint leaves juice. The low counts in the juices could be due to efficient
pasteurization and antimicrobial effect of mint and this implies that the microbial quality of the
product was satisfactory during 70 days of storage.

32
 5 CONCLUSIONS AND RECOMMENDATIONS

5.1 CONCLUSIONS
The purpose of this study was to evaluate the use of mint extract and hydrosol in flavoring and
improving the nutritional composition and antioxidant potential of passion fruit juice. Addition of
0-16% mint extract or hydrosol significantly lowers the sensory acceptability of passion fruit
juices. Up to 8% extract or hydrosol could be used to produce mint flavored passion fruit that is
acceptable to the consumer. Addition of mint extract to passion juice significantly improves the
vitamin C, beta-carotene and antioxidant activity of the juice. However, addition of mint hydrosol
(up to 8%) has no significant effect on vitamin C and beta-carotene content but increases the
antioxidant activity of passion fruit juice. Addition of 4-8% mint extract or hydrosol does not
significantly affect the shelf stability of passion fruit juice. Therefore, mint extract and hydrosol
can be used to flavor and also improve the nutritional and antioxidant properties of passion fruit
juice. Despite its negative effects on color, mint extract is better at improving the vitamin C, beta-
carotene and antioxidant activity of passion fruit juice.

5.2 RECOMMENDATIONS

Further research should focus on determining the effect of mint on other nutrients like iron,
calcium, total sugars, carbohydrates, protein and total energy of the passion fruit juice.
Physicochemical stability of the juice flavored with mint beyond 140 days should be studied.
Microbial stability of the juice beyond 70 days should also be studied. Sensory acceptability of the
juices during storage should also be studied to determine the time when the juice becomes un
acceptable to the consumers.

33
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 APPENDICES

Appendix I: Juices acceptable to consumers and selected for nutritional composition and shelf
stability studies

a b c d e

Appearance of passion fruit juices flavored with mint extract or hydrosol: control (a), 4% extract
(b), 8% extract (c), 4% hydrosol (d) and 8% hydrosol (e).

Appendix II: Total Plate Count of the mint flavored juices

Treatments/control 0 35 70
0%mint:100% Passion fruit juice ND ND ND
4% mint extract: 96% passion fruit juice ND ND ND
8% mint extract: 92% passion fruit juice ND ND <10cfu/ml
4% mint hydrosol: 96% passion fruit juice <10cfu/ml <10cfu/ml 17±2cfu/ml
8% mint hydrosol: 92% passion fruit juice ND ND ND
ND means not detected in the100 dilution

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Appendix III: Yeasts and Moulds of the mint flavored juices during storage
Sample/Storage time (days) 0 35 70
0%mint:100% Passion fruit juice ND ND ND
4% mint extract: 96% passion fruit juice ND ND ND
8% mint extract: 92% passion fruit juice ND ND ND
4% mint hydrosol: 96% passion fruit juice ND ND ND
8% mint hydrosol: 92% passion fruit juice ND ND ND
ND= Not detected

Appendix IV: Total coliforms in the mint flavored juices during storage
Sample/Storage time (days) 0 35 70
0%mint:100% Passion fruit juice ND ND ND
4% mint extract: 96% passion fruit juice ND ND ND
8% mint extract: 92% passion fruit juice ND ND ND
4% mint hydrosol: 96% passion fruit juice ND ND ND
8% mint hydrosol: 92% passion fruit juice ND ND ND

Appendix V ANOVA tables for sensory analysis of the mint flavored passion fruit juices
Mint extract
Analysis of variance (Appearance):
Sum of Mean
Source DF F Pr > F
squares squares
Model 4 170.217 42.554 16.138 <0.0001
Error 115 303.250 2.637
Corrected
Total 119 473.467

Analysis of variance (Color):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 196.367 49.092 17.391 <0.0001
Error 115 324.625 2.823
Corrected
Total 119 520.992

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Analysis of variance (Aroma):
Sum of Mean
Source DF F Pr > F
squares squares
Model 4 10.717 2.679 1.223 0.305
Error 115 251.875 2.190
Corrected
Total 119 262.592

Analysis of variance (Taste):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 22.217 5.554 2.049 0.092
Error 115 311.750 2.711
Corrected
Total 119 333.967

Analysis of variance (Acceptability):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 69.967 17.492 8.178 <0.0001
Error 115 245.958 2.139
Corrected
Total 119 315.925

Summary of ANOVA tables for Passion fruit juice flavored with mint extract
Appearance Color Aroma Taste Acceptability
R² 0.360 0.377 0.041 0.067 0.221
F 16.138 17.391 1.223 2.049 8.178
Pr > F < 0.0001 < 0.0001 0.305 0.092 < 0.0001

Mint hydrosol
Analysis of variance (Appearance)
Sum of Mean
Source DF F Pr > F
squares squares
Model 4 35.357 8.839 3.903 0.005
Error 110 249.130 2.265
Corrected Total 114 284.487

50
Analysis of variance (Color):
Sum of Mean
Source DF F Pr > F
squares squares
Model 4 5.617 1.404 0.686 0.603
Error 110 225.043 2.046
Corrected Total 114 230.661

Analysis of variance (Aroma):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 20.817 5.204 1.829 0.128
Error 110 313.043 2.846
Corrected Total 114 333.861

Analysis of variance (Taste):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 134.574 33.643 8.148 <0.0001
Error 110 454.174 4.129
Corrected Total 114 588.748

Analysis of variance (Acceptability):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 113.304 28.326 7.848 <0.0001
Error 110 397.043 3.609
Corrected Total 114 510.348

Summary of ANOVA tables for Passion fruit juice flavored with mint hydrosol

Appearance Color Aroma Taste Acceptability

R² 0.124 0.024 0.062 0.229 0.222
F 3.903 0.686 1.829 8.148 7.848
Pr > F 0.005 0.603 0.128 < 0.0001 < 0.0001

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Appendix VI: ANOVA tables for nutritional composition of the mint flavored passion fruit juice
Analysis of variance (Vitamin C mg/100ml of juice)
Source DF Sum of squares Mean squares F Pr > F
Model 4 28.140 7.035 20.750 0.003
Error 5 1.695 0.339
Corrected
Total 9 29.836

Analysis of variance (Beta-carotene µg/100ml of juice)

Source DF Sum of squares Mean squares F Pr > F
Model 4 4107.484 1026.871 58.978 0.000
Error 5 87.055 17.411
Corrected
Total 9 4194.540

Analysis of variance (Total antioxidants Concentration mg/100ml)

Source DF Sum of squares Mean squares F Pr > F
Model 4 6.266 1.567 10.762 0.011
Error 5 0.728 0.146
Corrected
Total 9 6.994

Summary of anova for total antioxidants, vitamin c and b-carotene

Total antioxidants
Concentration Vitamin C
mg/100ml mg/100ml of juice B-carotene µg/100ml of juice
R² 0.896 0.943 0.979
F 10.762 20.750 58.978
Pr > F 0.011 0.003 0.000
10.762 20.750 58.978
Samples
0.011 0.003 0.000

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Appendix VII: Anova tables for physicochemical properties of the juice during storage
ANOVA tables for pH at different days of storage
Analysis of variance (Day 0):
Sum of Mean
Source DF F Pr > F
squares squares
Model 4 0.024 0.006 3.261 0.113
Error 5 0.009 0.002
Corrected Total 9 0.033

Analysis of variance (Day 35):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 0.014 0.003 1.750 0.275
Error 5 0.010 0.002
Corrected Total 9 0.024

Analysis of variance (Day 70):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 0.030 0.008 2.500 0.171
Error 5 0.015 0.003
Corrected Total 9 0.045

Analysis of variance (Day 140)

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 0.026 0.006 0.929 0.515
Error 5 0.035 0.007
Corrected Total 9 0.061

Summary of ANOVA for pH

0 35 70 140
R² 0.723 0.583 0.667 0.426
F 3.261 1.750 2.500 0.929
Pr > F 0.113 0.275 0.171 0.515
3.261 1.750 2.500 0.929
Samples
0.113 0.275 0.171 0.515

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ANOVA tables for titratable acidity

Analysis of variance (day 0):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 0.004 0.001 14.391 0.006
Error 5 0.000 0.000
Corrected Total 9 0.004

Analysis of variance (day 35):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 0.013 0.003 28.917 0.001
Error 5 0.001 0.000
Corrected Total 9 0.013

Analysis of variance (day 70):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 0.005 0.001 5.994 0.038
Error 5 0.001 0.000
Corrected Total 9 0.007

Analysis of variance (day 140):

Sum of Mean
Source DF F Pr > F
squares squares
Model 4 0.003 0.001 16.746 0.004
Error 5 0.000 0.000
Corrected Total 9 0.003

ANOVA summary for titratable acidity

0 35 70 140
R² 0.920 0.959 0.827 0.931
F 14.391 28.917 5.994 16.746
Pr > F 0.006 0.001 0.038 0.004
14.391 28.917 5.994 16.746
Samples/Storage time
0.006 0.001 0.038 0.004

54
Appendix VII Sensory evaluation form
Sensory evaluation form for passion fruit beverage flavored with mint extract.
Date ……………………… Sex……………………………. Time …………………………….
You are requested to make sensory evaluation of mint flavored passion fruit beverages as presented
to you and evaluate their overall acceptability basing on the parameters shown below. Please grade
the samples on the scale of 1 to 9 by placing your score in the box next to the sensory parameter
under each sample in the box next to the sensory parameter under each sample in the table.
Rinse your mouth with water provided before tasting, re-tasting and in between tasting of the
beverages. Then evaluate the beverage in front of you.
Score the beverages using hedonic scale below
Like extremely 9
Like very much 8
Like moderately 7
Like slightly 6
Neither like nor dislike 5
Dislike slightly 4
Dislike moderately 3
Dislike very much 2
Dislike extremely 1

Quality Formulations
parameter 581 146 972 461 760
Appearance
Color
Flavor
Aroma
Taste
General
acceptability

Which formulation (only one) would you buy and why

………………………………………………………………………………………………………
………………………………………………………………………………………………………
General comments ………………………………………………………………………………..

Thank you for participating

55
Sensory evaluation form for passion beverage flavored with mint hydrosol
Date ……………………… Sex……………………………. Time …………………………….
You are requested to make sensory evaluation of mint flavored passion fruit beverages as presented
to you and evaluate their overall acceptability basing on the parameters shown below. Please grade
the samples on the scale of 1 to 9 by placing your score in the box next to the sensory parameter
under each sample in the box next to the sensory parameter under each sample in the table.
Rinse your mouth with water provided before tasting, re-tasting and in between tasting of the
beverages. Then evaluate the beverages in front of you.
Score the beverages using hedonic scale below
Like extremely 9
Like very much 8
Like moderately 7
Like slightly 6
Neither like nor dislike 5
Dislike slightly 4
Dislike moderately 3
Dislike very much 2
Dislike extremely 1

Quality Formulations
parameter 232 769 384 524 471
Appearance
Color
Flavor
Aroma
Taste
General
acceptability

Which formulation (only one) would you buy and why?

………………………………………………………………………………………………………
………………………………………………………………………………………………………
General comments………………………………………………………………………………….

Thank you for participating

56