Professional Documents
Culture Documents
Jun Ma
Faculty of Engineering
September 2015
PLEASE TYPE
THE UNIVERSITY OF NEW SOUTH WALES
Thesis/Dissertation Sheet
Banana pseudostems are crop waste, which cause economic loss and environment issues after harvest.
However, pseudostems are rich in dietary fibre and have health benefits. This study explored the chemical
composition (proximates, minerals, vitamins) as well as the digestibility and functionality of the carbohydrates.
Dietary fibre was estimated using three methods namely the established AOAC method, Gas Chromatography
and Nuclear Magnetic Resonance. As fresh banana pseudostems have limited shelf life, drying the stems using
cabinet dryer using several conditions such as, drying at 40 °C and 50°C with/ without blanching, were
compared with regard to drying time, colour as well as quality of the dried product in terms of the retention of
nutrients. Drying at 50 °C without blanching provided the whitest colour and shortest drying time. Thus the
optimum condition for drying was established at 50 °C without blanching based on nutrient retention. Musa
balbisiana and Musa acuminata banana pseudostems were used in this study. There was no significant
difference between protein, fat and carbohydrate content of banana pseudostems dried under different
conditions. Moisture content was significantly higher in banana pseudostems dried at 40 °C without blanching;
ash content was significantly higher in pseudostems dried at 50 °C without blanching. According to the
percentage of total dietary fibre and resistant starch, pseudostem dried at 40 °C with blanching had the lowest
digestibility. The neutral sugars in the non-starch polysaccharides were studied and compared with
commercially available natural dietary fibre supplements sold in Australia. Pseudostems had higher ratio of
soluble dietary fibre to insoluble dietary fibre, as compared to the commercial supplements. The main neutral
sugars in the pseudostems were glucose, mannose and xylose, while those in the commercial supplements
were xylose, arabinose and mannose, which had different functionality compared to pseudostem fibre. This
study is the first to demonstrate that banana pseudostem is a potential dietary fibre supplement, which may
bring health benefits to consumers and economic profits to the banana growers.
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archive and to make available my thesis or dissertation in whole or part in the
University libraries in all forms of media, now or here after known, subject to the
provisions of the Copyright Act 1968. I retain all proprietary rights, such as patent
rights. I also retain the right to use in future works (such as articles or books) all
or part of this thesis or dissertation.
I also authorise University Microfilms to use the 350 word abstract of my thesis in
Dissertation Abstract International (this is applicable to doctoral theses only).
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ABSTRACT
Banana pseudostems are crop waste, which cause economic loss and
fibre and have health benefits. This study explored the chemical composition
the carbohydrates. Dietary fibre was estimated using three methods namely the
Resonance. As fresh banana pseudostems have limited shelf life, drying the
stems using cabinet dryer using several conditions such as, drying at 40 C and
50C with/ without blanching, were compared with regard to drying time, colour
drying time. Thus the optimum condition for drying was established at 50 C
Pseudostems had higher ratio of soluble dietary fibre to insoluble dietary fibre,
which may bring health benefits to consumers and economic profits to the
banana growers.
Table of Contents
1. Introduction ........................................................................................................................ 1
i
2.4.2.1 Air properties ............................................................................................................................................ 27
ii
2.6.3 Vitamin B1 (Thiamin) ................................................................................................................. 56
iii
3.3.5.3 Procedure for sugar extraction ......................................................................................................... 71
iv
3.4.2 Procedure for determining swelling capacity (SWC) and solubility analysis ....101
pseudostem ...................................................................................................................................104
v
5.2.2.2 Method validation ................................................................................................................................ 121
5.2.2.3 Quantitative analysis of the vitamins in the banana pseudostem .................................. 124
6.2.3 Comparison between HPLC and Englyst methods for analysis of SDS, RDS and
vi
7. Effect of drying on the physicochemical properties of
vii
List of Tables
TABLE 2.1 THE TOP PRODUCERS OF BANANAS IN THE WORLD AND THE PRODUCTION IN AUSTRALIA IN 2012 ............ 4
TABLE 2.4 PROXIMATE COMPOSITION OF TENDER CORE OF BANANA PSEUDOSTEM FLOUR ............................................ 11
TABLE 2.7 CLASSIFICATION OF THE PRINCIPAL DIETARY CARBOHYDRATES (CUMMINGS ET AL., 1997)...................... 33
TABLE 3.14 CALIBRATION CONCENTRATION OF THE STANDARD SUGAR MIXTURE (MG/ L) ........................................... 91
viii
TABLE 3.21 EQUIPMENT USED FOR CI ANALYSIS ................................................................................................................. 102
TABLE 4.1 COLOUR OF DRIED GROUND BANANA PSEUDOSTEM (N= 3 ± SD) .................................................................. 107
TABLE 4.2 WATER ACTIVITY (AW) OF MUSA BALBISIANA AND MUSA ACUMINATA ......................................................... 109
TABLE 5.1 MEAN PROXIMATE CONTENTS OF MUSA BALBISIANA AND MUSA ACUMINATA.............................................. 110
TABLE 5.2 MEAN MINERAL CONTENT (MG/100 G) OF MUSA BALBISIANA AND MUSA ACUMINATA ............................ 115
TABLE 5.3 LINEARITY OF STANDARD CURVES AND SENSITIVITY FOR THE SEVEN B‐COMPLEX VITAMIN. ................... 121
TABLE 5.4 RESULT OF HPLC ANALYSIS OF B GROUP VITAMIN CONCENTRATIONS IN CERTIFIED REFERENCE MATERIAL
TABLE 5.5 INTER‐DAY (OVER A PERIOD OF 7 CONSECUTIVE DAYS) AND INTRA‐DAY (N=6) VALUES FOR THE B‐
TABLE 5.6 THE CONCENTRATION OF B‐COMPLEX VITAMINS (ΜG/G) IN BANANA PSEUDOSTEM DRIED IN DIFFERENT
TABLE 6.1 MEAN FRUCTOSE, GLUCOSE, SUCROSE AND TOTAL SUGAR CONTENT OF MUSA ACUMINATA PSEUDOSTEM
TABLE 6.2 RAPIDLY DIGESTIBLE STARCH, SLOWLY DIGESTIBLE STARCH, RESISTANT STARCH AND TOTAL STARCH
(ENGLYST METHOD & HPLC METHOD) OF MUSA ACUMINATA AND MUSA BALBISIANA PSEUDOSTEM (%, DB)
........................................................................................................................................................................................... 135
TABLE 6.3 RDS, SDS, RS AND TS CONTENTS IN SOME CARBOHYDRATE‐ CONTAINING FOODS ................................... 141
TABLE 6.4 RDS, SDS, RS AND TS CONTENTS IN SOME CARBOHYDRATE‐ CONTAINING FOODS ................................... 142
TABLE 6.5 MEAN SDF, IDF, TF OF MUSA BALBISIANA PSEUDOSTEM AND MUSA ACUMINATA PSEUDOSTEM ............ 146
TABLE 6.6: CHEMICAL SHIFTS AND PORTION OF SUGAR FRACTIONS IN THE BANANA PSEUDOSTEM DIETARY FIBRE
TABLE 6.7 GC DATA OF MONOSACCHARIDES IN STANDARD MIXTURE AND BANANA PSEUDOSTEM SAMPLES ............ 158
TABLE 6.8 CALIBRATION CONCENTRATION OF THE STANDARD SUGAR MIXTURE (MG/ L) ........................................... 159
TABLE 6.9 SENSITIVITY AND LINEARITY CHARACTERISTICS OF NEUTRAL SUGARS DETERMINATION IN THE PRESENCE
TABLE 6.10 INTER‐DAY (OVER A PERIOD OF 5 CONSECUTIVE DAYS) AND INTRA‐DAY (N=7) PRECISION AND
TABLE 6.11 DISTRIBUTION OF NEUTRAL SUGARS (G/ 100 G OF DRY SAMPLE) IN THE SOLUBLE, INSOLUBLE AND
ix
TOTAL FRACTIONS OF NSP FROM BANANA PSEUDOSTEM ........................................................................................ 165
TABLE 7.1 PHYSICOCHEMICAL PROPERTIES OF DRIED BANANA PSEUDOSTEM POWDER ............................................... 177
TABLE 7.2 PASTING PROPERTIES OF BANANA PSEUDOSTEM (BP40, BP40B, BP50, BP50B, BPA) MEASURED BY
x
List of Figures
FIGURE 2.1 BANANA PSEUDOSTEM ............................................................................................................................................... 6
FIGURE 2.7 SIMPLIFIED MECHANISM FOR THE HYDROXYLATION AND OXIDATION OF DIPHENOL BY PHENOLOXIDASE17
FIGURE 4.1 EFFECT OF PROCESSING ON THE DRYING RATIOS OF BANANA PSEUDOSTEM .............................................. 104
FIGURE 4.2 EFFECT OF SPECIES ON THE DRYING RATIOS OF BANANA PSEUDOSTEM ...................................................... 106
FIGURE 5.1 CHROMATOGRAPHY OF STANDARD THIAMIN AND 5‐MTHF WITH PH ADJUSTMENT (A) AND WITHOUT PH
FIGURE 6.1 EFFECT OF DRYING CONDITION ON THE RATIO OF FRUCTOSE, GLUCOSE AND SUCROSE OF MUSA
FIGURE 6.2 EFFECT OF BANANA SPECIES ON THE RATIO OF FRUCTOSE, GLUCOSE AND SUCROSE ................................. 133
FIGURE 6.3 EFFECT OF DRYING CONDITIONS ON THE RATIO OF RAPIDLY DIGESTIBLE STARCH (RDS), SLOWLY
DIGESTIBLE STARCH (SDS) AND RESISTANT STARCH (RS) OF MUSA BALBISIANA PSEUDOSTEM .................... 138
FIGURE 6.4 EFFECT OF BANANA SPECIES ON THE RATIO OF RAPIDLY DIGESTIBLE STARCH (RDS), SLOWLY DIGESTIBLE
FIGURE 6.5 PLOT OF HPLC METHOD VERSUS ENGLYST METHOD FOR TOTAL STARCH OF BANANA PSEUDOSTEM ... 143
FIGURE 6.6 TOTAL STARCH CONTENT OF BANANA PSEUDOSTEM DETECTED BY HPLC METHOD AND ENGLYST
FIGURE 6.7 A: 13C CP/MAS NMR SPECTRUM OF SOLUBLE DIETARY FIBRE OF DRIED BANANA PSEUDOSTEM
xi
B: 13C CP/MAS NMR SPECTRUM OF INSOLUBLE DIETARY FIBRE OF DRIED BANANA PSEUDOSTEM ............. 149
FIGURE 6.8 13C CP/MAS NMR SPECTRA OF COMMERCIAL PECTINS ............................................................................... 150
FIGURE 6.9 CP/MAS 13C NMR SPECTRUM OF CELLULOSE ISOLATED FROM SWITCHGRASS FROM LITERATURE ..... 151
FIGURE 6.10 1H NMR SPECTROSCOPY OF BANANA PSEUDOSTEM TOTAL DIETARY FIBRE EXTRACTION .................... 152
FIGURE 6.11 CHROMATOGRAMS OF DERIVATISED MONOSACCHARIDES IN A STANDARD SOLUTION (A) AND BANANA
FIGURE 6.12 CHROMATOGRAM OF (A) STANDARD WITH WRONG COMBINATION RATIO OF STANDARDS TO INTERNAL
STANDARD AND (B) SAMPLE WITH STALE AMMONIA SOLUTION ............................................................................. 157
FIGURE 7.1 XRD RESULTS OBTAINED FROM MUSA ACUMINATA (A), MUSA BALBISIANA (B) BANANA PSEUDOSTEM AND
xii
ACKNOWLEDGEMENTS
experimental and writing processes, I have been accompanied and supported by many
people. It is a pleasant aspect that I have now the opportunity to express my gratitude to
all of them.
First, I would like to thank my parents for their financial support to ensure my excellent
education abroad, their encouragement for me to learn new things and their mental
Jayashree Arcot and Dr. George Srzednicki, the School of Chemical Engineering,
inspired my love for food science and research. Without their excellent guidance,
I thank Paul Nicholson, Sydney Royal Botanic Garden, Australia, for banana
pseudostems supply. My sincere thanks to Mr. Lewis Adler for teaching me to use Gas
Chromatography and Dr. Maria Veronica Chandra-Hioe for teaching me to use High
Performance Liquid Chromatography. I thank them for their immense patience and
I would like to acknowledge the lab manager Camillo for his assistance and my friends
Yiqing Zhao, Ji Liang, Xin Sun, Yannie Pan, Na Wang and Kitty Tang for their help,
friendship and laughter in the labs. Finally, I would like to thank those closest to me
xiii
List of Abbreviations
5-MTHF 5-Methyltetrahydrofolate
Ara Arabinose
aw Water Activity
BD Breakdown
40 C without Blanching
50 C without Blanching
50 C without Blanching
DF Dietary Fibre
FA Folic Acid
Fuc Fucose
FV Final Viscosity
xiv
Gala Galactose
GI Glycemic Index
Glu Glucose
Spectrometry
IS Internal Standard
Man Mannose
NA Nicotinic Acid
NAM Nicotinamide
xv
NSP Non-starch Polysaccharide
POD Peroxidase
PV Peak Viscosity
PYR Pyridoxine
Rha Rhamnose
RIB Riboflavin
RS Resistant Starch
SB Set Back
TG Total Glucose
THF Tetrahydrofolate
THI Thiamin
TS Total Starch
TV Through Viscosity
xvi
UV Ultraviolet
Xyl Xylose
xvii
1. Introduction
production, it is in the second place after citrus, accounting for about 16% of the
world’s total fruit production (Deharveng et al., 1999). There are two wild
species of banana, namely Musa acuminata and Musa balbisiana. Almost all
modern edible parthenocarpic bananas come from these two species (Robinson
& Sauco, 2010). Therefore, these two species of bananas were chosen for this
study. The stem of the banana plant, which is also called pseudostem produces
(Anhwange et al., 2009). This crop generates a large amount of residue, due to
the fact that each plant produces only one bunch of bananas. After the harvest,
the bare pseudo-stem is cut and usually left on the plantation or burned, which
could ultimately cause environment issues (Cordeiro et al., 2004). Thus the
recent years. The banana pseudostem has been used as material for paper,
furniture and forage (Buragohain et al., 2010; Umaz et al, 2005). Moreover, it
has been reported that these banana waste materials are rich in minerals and
nutrients, especially dietary fibre (Aziz et al., 2011). However, little is known on
potentially be used more in food rather than in other industries. The exploitation
1
environment and increase its economic value. This study focused on efficient
fibre components.
2
2. Literature Review
The origin of the banana plant is complex because of the nature of the banana’s
domesticated banana was grown in New Guinea around 8,000 BC. From New
and then radiated widely across the tropics. It took probably two millennia for
Malaysia. Plantains may have been grown in eastern Africa as early as 3,000
Indian traders travelling through the Malaysian region had tasted the fruit and
brought plants back with them in 600 BC. In 327 BC, when Alexander the Great
and his army invaded India, he discovered banana crop and tasted the fruit in
the Indian Valley. Then he introduced this new discovered crop to the Western
By 200 AD banana had spread to China and grew only in the southern region of
China at that time. The Chinese never really popularised this fruit until the 20th
Century as they were considered to be a strange and exotic alien fruit. At about
Table 2.1 shows the production of banana worldwide and the production in
3
Australia. Foulkes et al. (1978) indicated that banana fruit occupied 25% of the
total banana plant weight, while banana pseudostem occupied 61% of the
weight (Pérez & Fujita, 1997). This means every year approximately 600,000
produced annually.
Table 2.1 The top Producers of bananas in the world and the production in Australia in 2012
not contribute a major share. In Australia, bananas are grown in both tropical
the farming practices, the geographical location of banana farms, the size and
4
type of farms that grow bananas, the varieties of bananas grown and their
Table 2.2. All states in Australia have banana production, but Queensland
dominates the production. All fresh bananas available in Australia are grown
Banana (Musa sp) belongs to the Musaceae family. Over 70 species of Musa
were recognized by the World Checklist of Selected Plant Families, but only few
species are edible. There are two wild species of banana, including Musa
bananas come from these two species (Valmayor et al., 1999). As a result, this
5
study focused on these two species. The hybrid of Musa acuminata and Musa
Banana pseudostem (Fig 2.1 & Fig 2.2) is the stem of banana plant; it produces
a single bunch of banana before dying and then is replaced by new pseudostem
(Anhwange et al., 2009). Since each plant produces only one bunch of bananas
and cannot be used for the next harvest, this agricultural activity generates a
large amount of residue (Cordeiro et al., 2004). It has been reported that
about 16% of the world’s total fruit production (Mohapatra et al., 2010).
cut and left behind as waste worldwide, which ultimately causes contamination
of water sources as well as can affect the environment and health of living
6
Figure 2.2 Banana plant (Source: Photograph taken by the author)
Hence, the exploitation of waste banana pseudostems into products could
Figure 2.3 shows the structure of banana plant. The plant is normally tall and
fairly sturdy, as a result is often mistaken for a tree. However, the trunk of the
banana plant is actually a false stem or pseudostem (Stover & Simmond, 1972).
The pseudostem is normally 5 to 7.6 meters tall (varies from species to species)
growing from a corm (Nelson et al., 2006). As shown in Figure 2.4, the
pseudostem consists of a tender core and several outer sheaths. The tender
core (Fig 2.5) inside the pseudostem carries the immature inflorescence until
7
Figure 2.3 The structure of banana plant
8
Outer sheaths
banana
9
2.2.3 Nutritional value of banana pseudostem
Banana pseudostem has very high content of dietary fibre. It has caught the
attention of food scientists in recent years. It could be used more in food rather
than in the feed industry. Aziz et al. (2011) and Bhaskar et al. (2011)
Protein 2.5
Fat 1.7
Starch 27.3
Ash 0.3
Moisture 15.1
10
Table 2.4 Proximate composition of tender core of banana pseudostem flour
Moisture 8.8
Fat 1.2
Protein 3.5
Ash 10.1
The values from these two studies are considerably different to each other. One
possibility is that some of the nutrients were underestimated from the study of
Bhaskar et al. (2011). For example, the ash value of that study was 0.3%. In
contrast, Ho et al. (2012) reported that the total ash was 6.8% and mineral
zinc and manganese was 3.1%. The total mineral value detected by the study
was even more than 0.3%. Hence, it is reasonable to believe the ash measured
that the methods used in the two studies were different. The total carbohydrate
obtained from the study of Bhaskar et al. (2011) was the sum of sugar, starch
and dietary fibre, while the total carbohydrate was calculated as 100% minus
moisture, fat, protein, ash and crude fibre in the study of Aziz et al. (2011). The
study of Aziz et al. (2011) has some limitations. The crude fibre is part of total
11
would underestimate the total carbohydrate value. The value of sugar, starch,
dietary fibre could not be shown from this kind of calculation. The studies on the
may all cause the differences in the nutrient values of the banana pseudostem.
For example, Saravanan and Aradhya (2011) claimed that total phenolics and
cultivars are different. They may range from 7.58 to 291 mg gallic acid
sugar content and decrease in starch. Marlett & Vollendorf (1994) compared
two methods, the AOAC method (Prosky et al., 1987) and Uppsala method
(Theander et al., 1990), to determine dietary fibre content in fruits. They found
that the AOAC method always had greater concentration of dietary fibre than
Uppsala method, and the two data sets were significantly different.
2.2.3.2 Minerals
Minerals play an important role in maintaining proper function and good health
in the human body (Bhat et al., 2010). Deficiency of minerals in the diet is
12
pseudostem are shown in Table 2.5. The mineral content will be affected by
(Ho et al., 2012; Happi-Emaga et al., 2007; Lahav et al., 1985). For instance,
tissues.
Sodium 444.1
Potassium 944.1
Calcium 1335.3
Magnesium 255.0
Phosphorus 137.8
Iron 3.3
Zinc 8.1
Manganese 1.3
and paper industry. Buragohain et al. (2010) claimed that banana pseudostem
could be used as an important staple food for pigs in banana producing areas.
They can be used as feed material in both fresh and sun-dried forms and both
13
could also be used to produce the glue used in the manufacture of cartons.
Umaz et al. (2005) reported that due to the fact that the fibre of banana
pseudostems is widely recognized for its good qualities vs. synthetic fibres, it is
used for making apparels, garments and home furnishing. Banana pseudostem
has been seen as a kind of vegetable in some countries. For example, in India
and Malaysia, the fresh tender core of banana pseudostem is cooked and
Australia is rare.
This browning reaction will change sensory properties and decrease nutritional
acceptance. Since the browning reaction can affect the quality of food products
storage, the following paragraphs will review the causes of browning reactions
Browning reaction is one of the most important and common reactions that
takes place during food processing and storage. There are three main causes
14
2.3.1.1 Enzymatic browning
It is estimated that enzymatic browning is the main cause in plant material and
one of the most important colour reactions that affects the quality of fruits and
vegetables, such as apple, banana and potato. After harvest, the cell wall of
plant material is disrupted and the enzymes form brown or sometimes yellow,
tyrosinase that are widely distributed from bacteria to mammals and are
defense mechanism against plant pathogens. They make the tissues brown by
are oxidoreductases that are able to oxidize phenol compounds. They act by
shown in Figure 2.6. PPOs catalyze the hydroxylation to the o-position adjacent
form high molecular weight polymers, such as melanoidin, which display brown
and dark pigments in injured vegetable and fruit tissues (Mai & Glomb, 2013).
Figure 2.7 describes the simplified mechanism for the hydroxylation and
15
oxidation of diphenol by phenoloxidase. PPO, as a bi-functional enzyme,
containing copper in its structure, has been described as oxygen and four
expense of peroxides. Although they are highly specific for hydrogen peroxide,
Peroxidases are widely distributed, especially in plants, but it is implied that they
vegetables following mechanical stress (Caballero et al., 2003). Thus, PPOs are
16
Figure 2.6 Polyphenol oxidase pathway
(Source: Corzo-Martínez et al., 2012)
Figure 2.7 Simplified mechanism for the hydroxylation and oxidation of diphenol by
phenoloxidase
Source: Corzo-Martínez et al., 2012
17
(Manzocco, 2000). These reactions can promote nutritional changes such as
reaction. It occurs as a result of the reaction between free amino groups from
amino acids, peptides or proteins and the carbonyl group of a reducing sugar. In
this stage, the reaction loses a molecule of water and forms glycosylamines.
(Lee & Whitaker, 1995). Figure 2.8 shows the outline of the chemical pathway
of Maillard reaction.
18
Figure 2.8 Outline of the chemical pathway of Maillard reaction
plant organs which are rich in oxidizable phenols such as fruits and vegetables
and decrease their economic returns in the food industry, measures should be
browning. There are various methods that can be used for browning
19
prevention. They can be divided into two categories—physical methods and
this section.
Although there are several methods to prevent enzymatic browning, they have
the same principle, which is to reduce or inactivate the enzyme, and thus to
Freezing
Freezing (below -18°C) is estimated to block colour changes due to the fact that
enzymes such as PPOs could be inactive under this condition. However, when
the temperature rises, browning starts again, and will be even more visible if the
cellular structures of the plant organ have been severely damaged by freezing,
chemical peeling and slicing (Caballero et al., 2003). Cano et al. (1997) support
Blanching
to the fact that blanching could reduce the microbial load and inactivate
20
deleterious enzymes, such as PPOs and PODs. Castro et al. (2008) implied
significant inactivation of PPO and POD (96-100%). Bahçeci et al. (2005) also
agreed that blanching could inactivate enzymes. They found that a blanching
There are two major methods that are widely used in the food industry for
blanching. One is hot water blanching and the other is steam blanching. Hot
from steam to inactive enzymes. Compared with hot water blanching, steam
blanching uses less energy and water. Johnson (2011) illustrated that when
steam blanching was applied directly to the food products, it used approximately
half the amount of water vs. that used in water blanching. It could save
approximately $4000- $8000 in energy cost by steam blanching one million lbs
(454 metric tons) of carrots or peas than water blanching. Moreover, steam
blanching achieved better nutrient retention than water blanching. The nutrient
loss of steam blanching is only one third of hot water blanching (Lee &
Whitaker, 1995). When food enters hot water, some soluble nutrients such as
soluble vitamins may dissolve in the hot water and the nutrients in the hot water
blanched food could be lost. Because steam blanching minimizes the leaching
of soluble solids, this method could leave more nutrients and natural
21
sugars in food products, thus it could improve flavour retention and colour
retention to produce a final product with better flavour, texture and colour than
sensory (texture, taste, flavour and colour) and nutritional quality attributes
(Castro et al., 2008). Jackson et al. (1996) blanched whole green bananas at
50, 60,70,80,90 and 100 °C for 2, 15 and 30 minutes, then fried peeled and
would have negative effects on crispness. Castro et al. (2008) stated that
more severe. The higher the temperature and longer treatment time, the lower
the ascorbic acid content in the samples. The retained ascorbic acid content in
green and red peppers was 45% and 30% respectively. Therefore, blanching
temperature and time should be well controlled. The optimal temperature and
duration time is the condition that could both inhibit enzyme activities and
are also used in the food industry to protect the colour of foodstuff. Palou et al.
22
Caballero et al. (2003) stated that browning can also be prevented by keeping
the food products in low oxygen atmospheres. Nowadays, food industry uses
nitrogen provides better protection of the original flavour and aroma. Segovia-
atmosphere for 24 h. They found that the browning of bruised areas of the
olives was very similar to that suffered by healthy olives maintained in the air,
which meant the N2 atmosphere was able to prevent the browning. However, N2
atmosphere is not suitable for any food. There is evidence showing that
2012).
are used to inhibit browning (Queiroz et al., 2011). Some of the compounds
could also prevent both enzymatic and non-enzymatic browning. Despite the
fact that some compounds, such as sulfur dioxide, have significant effects on
colour and flavour preservation, they cause safety issues and thus are
most countries allows only ascorbic acid and its derivatives, sodium chloride
and, within stricter limits, citric acid (Caballero et al., 2003) to be used for this
23
synergistic effects will be reviewed in the following paragraphs.
It has been reported that ascorbic acid showed strong inhibition of PPO,
causing full enzyme inactivation even at low concentration. Ascorbic acid with
Ünal (2006) proved that ascorbic acid at 0.2 mM and 0.8 mM resulted in 99%
and 100% inhibition of banana PPO, respectively (Queiroz et al., 2011). Täufel
apple pieces, whereas Pizzocaro et al. (1993) argued that sodium chloride in
concentrations between 0.2 g/L to 1 g/L activated PPO and 1 g/L even
than that of single compounds. Pizzocaro et al. (1993) studied the synergistic
effect of ascorbic acid and citric acid plus ascorbic acid and sodium chloride.
They discovered that both citric acid and sodium chloride increased the
inhibiting effect of ascorbic acid. They concluded that compared with citric acid,
sodium chloride is more efficient. Adding 0.5 g/L of sodium chloride instead of 2
g/L of citric acid to 10 g/L of ascorbic acid showed that the PPO inhibition was
24
2.4 Drying
Drying is one of the most effective methods used to preserve fruit and
vegetables, because fruits and vegetables have high moisture and this method
can remove the moisture in food by evaporation. The rate of microbial activity
and other deteriorative reactions in fresh fruit and vegetables therefore can be
the physical properties of the fibre matrix and affect the hydration properties as
well (Dhingra et al., 2012). Different drying conditions may also cause a
difference in food quality. If the drying time is too long or the drying temperature
is too high, the quality of the food including nutritional values, colour, taste etc.
chosen during food preservation. In this section, the principle of drying, drying
curve, factors that will affect drying rate as well as effect of drying conditions will
be presented.
Drying is a process that utilizes the dry air to evaporate bound water from inside
the fresh food into the atmosphere. Breaking water bonds, releasing and
convective (warm air), contact (cooled surface), radiative (infrared rays), and
excitation (microwave) energies (Hui et al., 2006). There are two basic
25
mechanisms involved in the drying process; one is the migration of moisture
from the interior of the food to the surface and the other is the evaporation of
moisture from the surface to the surrounding air. When hot air is blown over a
wet food, water vapor diffuses through a boundary film of ambient air and is
carried away by the moving air (Figure 2.9). A water vapour pressure gradient is
established from the moist interior of the food to the dry air, thereby most
constant rate period and falling rate period (Figure 2.10). Initial rate period is a
short initial settling down period as the surface of the food heats up to wet bulb
temperature. At this stage, the drying rate keeps increasing. Constant rate
period is the stage where the water moves from the interior of the food at the
same rate as it evaporates from the surface. This step stays till the critical
moisture content is reached. The unbound water is removed from the food
product. The evaporation is not dependent on the solid matrix and does not end
26
until water from the interior is no longer available at the food surface
(Geankoplis, 2003). Falling rate period is the last and the longest part of drying
operation. The drying rate falls down slowly until the moisture content in the
product reaches the humidity of the air in the dryer. This moisture content of the
The properties of the air flowing around the food being dried are a major factor
in determining the rate of drying. Air temperature, air humidity and air velocity
are three key properties of air that affect the drying rate (Fellows, 2002). Low
27
temperatures promote drying rate. Sacilik and Elicin (2006) studied the
apple slices. They stated that an increase of drying air temperature from 40 to
Meanwhile, Doymaz (2007) proved that the increase in the air temperature in
drying, because fresh air passing over a wet food helps removal of moisture.
The drying rate is affected by different physical properties of food such as the
example, for foods that have the same volume, the one in small pieces and
larger surface area has quicker drying rate than the one with small surface area
Other factors such as blanching, freezing, osmotic pretreatment will also affect
the drying rate. Agarry et al. (2005) concluded that freezing and increasing
freezing time could significantly increase the drying rate and therefore decrease
the drying time of potatoes. Similarly, Dandamrongrak et al. (2003) claimed that
blanching combined with freezing were 22.4 and 27.2 hours respectively. This is
28
2.4.3 Drying models
Various drying models were developed for different foods owing to their
models are very useful in prediction of drying times under particular process
conditions; therefore they are widely used in the drying process. Four typical
two-term exponential model of drying solids and some equations for drying are
is moisture ratio
Wo Wf
MC (2-1)
Wo
Where, Wo and Wf is the initial and final weight of the food material (g).
29
• Moisture ratio (MR); equation (2-2)
(2-2)
moisture content of the sample at each moment (M) and initial moisture content
enhance product quality, such as colour and flavour. Although high drying
temperature could improve the drying rate and decrease drying time in food
processing, it also has an adverse effect on food quality. It has been reported
that lower drying temperature should maintain original colour of fresh apple
slices (Sacilik & Elicin, 2006). However, the studies on the influence of drying
2.5 Carbohydrates
Carbohydrates, the main source of the human diet, represent the primary
30
energy source, contributing to nearly 55-70% of the total energy consumption
precursors for aroma and coloring substances in the food industry, especially in
thermal processing (Belitz et al., 2009). For instance, the Maillard reaction and
into the classification of carbohydrate. The 3 main groups are then subdivided
on the basis of the types of monosaccharide that are consistent, with a view to
(Table 2.7) not only presents the chemistry of the carbohydrates, but also
31
2.5.1.1 Mono- and di-saccharides
Free sugars are the simplest form of carbohydrate, which are present in fruits
and vegetables. Mono-saccharides, such as glucose and fructose, are the basic
According to the chain length of the mono-saccharides, they can be divided into
usually the most abundant disaccharide in the diets (Asp, 1995), which consists
rapidly digested and absorbed in the small intestine and provide a ready source
of energy.
32
Table 2.7 Classification of the principal dietary carbohydrates (Cummings et al., 1997)
and can be only partially digested by humans. The most commonly known
oligosaccharides are mainly polymers of fructose and galactose. They are found
in plant seeds, especially legumes such as beans and peas. According to the
portal blood to the liver and eventually to the systemic circulation. The non-
digestible oligosaccharides pass into the large bowel as they are eaten
2.5.1.3 Polysaccharides
which can be further divided into starch and non-starch polysaccharides (NSP).
2.5.1.3.1 Starch
Starch is the major carbohydrate in the human diet, which occupies 80-90 % of
all polysaccharides eaten (Cummings et al., 1997). The starches are storage
34
Figure 2.11 Bioavailability of digestible and no-digestible oligosaccharides
35
According to the chemical structure, starch is a mixture of two large
anhydroglucose units and this molecular size varies depending on the plant
1, 6 linked D-glucose and its molecular weight exceeds 107 (Gallant et al.,
1992). Most starches have amylose content of about 25% and amylopectin
content of about 75% (Bemiller and Whistler, 1996). Starches that are made up
Molecular structure Linear (- 1,4) Branched (- 1,4 and -1,6)
amylase
There are three crystalline structures of starch granules in the plant, namely A,
36
formed in cereal starches, where water molecules are located between starch
molecules forming double helices. Potato and other tuber starches mostly have
the B structure; the double helices are packed densely in a hexagonal pattern
with water molecules only inside this structure. C pattern is mostly found in
In 1992, Englyst et al. further divided starch into rapidly digestible starch (RDS),
slowly digestible starch (SDS) and resistant starch (RS), according to their
Table 2.9 In vitro nutritional classification of starch (Source: Englyst et al., 1992)
small intestine
RS
seeds
starches
RDS = rapidly digestible starch; SDS = slowly digestible starch; RS= resistant
starch; RS1 = physically inaccessible starch; RS2= resistant starch granules;
RS3 = retrograded starch; RS4 = chemically modified starch
37
It has been once assumed that starch hydrolyzed and absorbed more slowly
than sugars, because they have larger molecular sizes than sugars. However,
RDS is rapidly digested and gives raise to blood glucose responses similar to or
even greater than sugars (Wolever & Miller, 1995). On the other hand, SDS and
RS can slow gastric emptying; reduce the glycemic index and insulin
glucosidic linkage, which are mostly the plant cell walls. They are found in all
cereals, vegetables and fruits (Blackwood et al., 2000; Englyst et al., 1994).
According to their solubility, NSP can be divided to soluble NSP and insoluble
NSP. The NSP cannot be digested in the small intestine because human
digestive enzymes can only cleave α - (1→ 4) glucan bonds (Nielsen, 2010).
The NSP offers health benefits as well. They are fermented by microbes in the
large intestine (Bhaskar et al., 2011). Due to their high water binding capacity,
NSP play an important role in providing bulk to gut contents and allow easy
passage through the human intestine. Thus, they play a crucial role for the
correct functioning of the digestive system (Nielsen, 2010; Kumar et al., 2012).
38
diabetes, are by far the leading causes of mortality in the world, representing
dietary fibre may be the constituents that could protect human beings from
these modern diseases. Dietary fibre has been shown to have numerous
benefits, which are not easily disputed like improving intestinal function,
and resistant starch include weight reduction, satiety, regulating glucose and
tumor necrosis factor alpha in obesity and diabetes (Cui & Roberts, 2009).
2.5.2.1 Hyperlipidemia
People now pay more attention to this disease since it could cause other
39
low density lipoproteins (VLDL), low density lipoproteins (LDL), intermediate
It has been reported that the proper diet treatment can preserve and reduce the
from hyperlipidemia. Kim et al. (2003) studied the effect of resistant starch on
hyperlipidemia actions in diabetic rats. They found that the total plasma lipid
treated rats than the diabetic control ones. Furthermore, some forms of dietary
fibre are also believed to have positive effect on lowering blood lipids, total
cholesterol and LDL cholesterol (Leeds, 2005). Soluble dietary fibre like pectin,
guar gum and gum may lower both serum cholesterol and triglycerides (Dhingra
et al., 2012). Brown et al. (1999) studied the cholesterol lowering effects of
dietary fibre. The results showed that intake of 2 to 10 grams of soluble dietary
fibre every day was associated with small but significant fall in total cholesterol
[-0.045 mmol/L· g soluble fibre (95% CI: -0.054, -0.035)] and LDL cholesterol [-
0.057 mmol/L · g (95% CI: -0.070, -0.044)]. Leeds (2005) claimed that intake of
food products providing more than 3 g of soluble fibre every day had a greater
40
(insulin-dependent or type I diabetes) or impaired responses of tissues to insulin
(Bender, 2009). Diabetes has been a growing public health problem worldwide.
(Cui & Roberts, 2009). In Australia, the prevalence of diabetes more than
doubled from 1.5% to 5.1% of Australians from 1989 to 2008. A large body of
evidence has demonstrated that the presence of diabetes doubles the risk of a
with death from cancers of the liver, pancreas, ovary, colorectum, lung, bladder
and breast (Dong et al., 2011). Numerous reports claimed that dietary therapy
preventive foods are urgently needed to reduce the huge burden of diabetes.
glycemic response from a standard food, usually glucose or white bread. Low
41
classification of carbohydrate foods based on the absorption rate of
responses of both single and mixed meals (Miller, 1994). Thus, it has now been
Epidemiological studies implied that low GI diets may reduce the risk of
believed to resist the raising of blood glucose after meal. One hypothesis
NSP, pectins and gums, may affect glucose and lipid metabolism. The major
acids, which are rapidly absorbed by the colonic epithelia with significant
quantities of acetate and propionate entering the portal blood stream. The
acetate may lower plasma nonesterified fatty acids levels. It has been implied
that elevated plasma nonesterified fatty acids levels may contribute to the
impairment of insulin action (Cho, 2001). Thus the effect of acetate to lower the
level of plasma nonesterified fatty acid may prevent the insulin action from
impairing.
Behall et al. (2006) stated that resistant starch has the effects to lower the
42
plasma glucose and insulin responses of both normal and overweight women.
On the one hand, some food scientists support the idea that insoluble dietary
fibre could decrease the risk of type II diabetes. Meyer et al. (2000) studied the
Iowa women aged 55-69 for 6 years. The result showed that, whole-grain, total
dietary fibre and cereal fibre had strong inverse associations with incidence of
diabetes after adjustment, which indicated a protective role of total dietary fibre
found that the soluble fibre had no significant association with type II diabetes;
therefore fibre from fruit and vegetables was unrelated to diabetes risk. The
twenty-five foods and their GI, had similar results. The result of Wolever’s study
showed that total dietary fibre was significantly related to GI and soluble fibre
On the other hand, some researchers claim that soluble fibres had greater
studied the effect of increasing the intake of dietary fibre on glycemic control in
twelve men and a woman patient with type II diabetes. They found that an
soluble fibre, significantly improved glycemic control and decreased the degree
type II diabetes.
43
Coronary heart disease is the major cause of death in most western countries.
Healthy eating for CHD prevention is a hot issue in the latest decade. High
carbohydrates reducing the risk of CHD improve blood lipid profiles by lowering
blood pressure and improving insulin sensitivity and fibrinolytic activity (Pereira
et al., 2004).
During the past decades, there were numerous key epidemiological studies to
prove that dietary fibre could prevent CHD. In 1999, Wolk et al. indicated that
higher fibre intake, especially cereal dietary fibre intake, reduced the risk of
CHD in women. They studied 68782 women aged 37 to 64 years for ten years
and concluded that women in the highest quintile of cereal fibre intake had a
34% lower risk of total CHD compared with those in the lowest quintile. Pereira
et al. (2004) followed up 91058 men and 245186 women over 6 to 10 years and
corrected total dietary fibre. They found a 14% decrease in risk of all coronary
studies by Pietinen et al. (1996) of Finnish men; studies of Ludwig et al. (1999)
44
material under specified conditions of temperature, time soaking, and duration
and speed of centrifugation (Elleuch et al., 2011). Dietary fibre holds water by
adsorption and absorption. WHC has a significant effect on fecal output and
property that has been widely studied in food, since they are associated with
food quality. The WHC of fibre differs between fibre sources. Table 2.10 shows
the water holding capacity of some foods and food by products. According to
Table 2.10, cereal dietary fibre presents lower water holding capacity than fibre
from fruit by-products and algae. For WHC purposes, dietary fibre from fruit by-
45
Table 2.10 Water holding capacity of some fibre
Algae 20.8
(Source: Elleuch et al., 2011; Aziz et al., 2011; Grigelmo-Miguel et al., 1999)
46
2.5.3.2 Swelling capacity (SWC)
material that can be absorbed. Food materials high in dietary fibre are always
associated with higher swelling capacity. For example, the swelling capacity of
the tender core of banana pseudostem, whose dietary fibre content is 47.98, is
13.82 grams swollen granules per gram of dry matter (Aziz et al., 2011). The
temperature (Elleuch et al., 2011). Raghavendra et al., (2006) claimed that with
the reduction of the particle size of coconut milk residue from 1127 to 550 μm,
the swelling capacity increased from 17 to 20 mL/g. They guessed that this
might be due to the increase in theoretical surface area; total pore volume and
structural modification.
2.5.3.3 Viscosity
Viscosity (g), which is the resistance to flow, is defined as the ratio of shear
stress (C) to shear rate (c). It is one of the most important physical properties of
solution increases. For example, long chain polymers, such as the gums (guar
gum, tragacanth gum) bind significant amount of water and exhibit high solution
47
increasing the viscosity of a solution (Kumar et al., 2012; Elleuch et al., 2011).
DFs, which have the ability to form viscous solutions or gels, can change the
rheology of the intestinal contents, and are known to produce local responses
along the gastrointestinal tract. Dietary fibre interacts with the intestinal brush
border and thickens the rate-limiting unstirred water layer of the mucosa
Moreover, high digested viscosity delays gastric emptying and feed transit time
growth in the intestine (Kumar et al., 2012). Since the characteristic of DFs is to
Additionally, dietary fibres could affect the rheological behavior and enhance
viscosity development when added into food products. Soukoulis et al. (2009)
studied the enrichment effect of ice cream with dietary fibre. They concluded
that dietary fibre significantly increased viscosity development when mixed with
ice cream. They inferred that the enhancement is due to the contribution of
insoluble fibres to the increase of total solids, which affects the three-
dietary fibre into rice starch to detect its influence on the physicochemical
characteristics of rice starch. They found that addition of dietary fibre (at 5%
wt% on dry starch basis) into Taichung Sen 10 rice flours could increase peak
2.5.3.4 Gelatinization
Undamaged starch granules are tightly packed and generally insoluble in water
48
because of the collective strength of the hydrogen bonds binding the chains
together (Coultate, 2002). Since the starch/ water system undergoes an order-
disorder transition during heating, the textural and digestive properties of food
(Liu, Lelievre & Ayoung-Chee, 1991). Water begins to be imbibed when the
temperature. When the water is 30 °C, the starch granules are intact and
absorb some water. When the temperature reaches 40 °C, more surface water
is adsorption, hydrogen bonds start to loosen slightly and water may enter some
adsorbed onto the surface and absorbed into granule. At this stage, some of the
amylose can move into water around granules and granule structure further
opens and swell. When the temperature is 60 °C, water forms H bonds with
amylose and amylopectin. As a result, the amount of free water outside the
granule decreases, the amount of amylose outside the granule increases and
the granule expands. The starch solution becomes more translucent and
the starch. Shorter amylose chains are more likely to leave granule and the
temperature raising closes 100 °C, granules may implode and fragment, hence
2.5.3.4.1 RVA
49
viscometer with ramped temperature and variable shear capability optimized for
testing the viscous properties of starch, grain, flour and foods. It provides a
viscograph that can assess the starch viscosity and gelatinization properties.
The RVA was developed by Newport Scientific, Australia, with the aim of
offering starch analysis using smaller sample size as well as a choice of short
and long test time (Haase et al., 1995). The RVA has been widely used in
research into food products like wheat, potatoes, cereal, maize and rice (Haase
et al., 1995; Almeida-Dominguez et al., 1997; Mijland et al., 1999; Yun and
(Prosky et al., 1987). The residue of the enzymatic digestion is used for
insoluble dietary fibre measurement after correction for protein and ash in the
aqueous ethanol.
Enzymatic –gravimetric method is one of the most widely used methods for
dietary fibre analysis. It could provide the amount of soluble, insoluble and total
dietary fibre in dried samples. However, the limitation of the gravimetric method
buffers and samples are insoluble in alcohol, as a result, the amount of ash is
50
Moreover, this method is based on a definition of dietary fibre as the sum of
are not measured by this method, which appears to underestimate the value of
fractions in the dietary fibre cannot be identified in this method (Caprita &
Caprita, 2011).
polysaccharide (NSP) has evolved from the principles laid down by Southgate
in 1969 (Englyst et al., 1994). The enzymatic removal of starch and some
as the sum of the constituent sugars released by acid hydrolysis. This method
gives values for individual monosaccharides and uronic acids released from
NSP.
Furthermore, it quantifies the sugar fractions in the NSP, which indicates the
51
based on selectivity by volatility and requires derivatization; however
utilizes the spin properties of certain atomic nuclei, the most prominent being 1H
13
and C. The spins will align either parallel or anti-parallel to the magnetic field
when exposed to a strong magnetic field. The strength of the magnetic field is
difference in energy. As a result, spins can be excited from the low to the high
weaken the magnetic field as well and thus give characteristic deviations in the
Moreover, the signal of different sugar fractions may overlap, no unique signal
can be used to identify and quantify the fractions in the dietary fibre.
52
prediction of the component in new samples. Since this method relies on the
NIR analysis is very rapid, requires little or no sample preparation, and there is
no creation of chemical waste (Kays et al., 1999). However, this analysis can
suitable for large scale determination of dietary fibre in the industry rather than
Vitamins can be divided into two major groups, including water-soluble and fat-
soluble vitamins. The major part of the water-soluble vitamins comprises the B-
is required daily, they play very important roles in our health. Deficiency of the
anemia (Aminoff et al., 1999; Burch et al., 1950; Chindarkar et al, 2014). This
chapter will review some of the B-complex vitamins, their requirements, excess
and deficiency, sources from food matrix, bioavailability and stability. Moreover
2.6.1Vitamin B3 (Niacin)
53
Niacin (known as vitamin B3) is the generic descriptor for two vitamers (nicotinic
acid and nicotinamide), which are colorless and odorless and both based on a
group (Rose-Sallin et al., 2001). The two compounds have similar absorption
spectra in water with absorption maximum at near 260 nm. These two forms of
niacin have equal biological activity and are easily inter-convertible. They are
unaffected by air, oxygen, light, acid, alkali and heat in the dry state and in
neutral aqueous solution (Combs, 2008; Ball, 2005; Eitenmiller et al., 2007).
Hence, in food processing, niacin would not be lost during storage, cooking and
heating. However, since niacin is freely soluble in boiling water (Ball, 2005), it is
fifty percent of the available niacin is lost when combining soaking and cooking
chicken (CNF, 2010). However, its bioavailability in natural foods is often low.
For example, it has been found that natural cereal grains contain a significant
amount of niacin, but 85% - 90% of the niacin is present in chemically bound
form, which cannot be absorbed or utilized. Niacin often occurs in food, bound
peptides via amide linkage between amino group and peptide as niacinogen. In
54
hydrolysis is often used to liberate the nicotinic acid from the bound forms. This
allows the determination of accurate total content of niacin, but at the same time
can overestimate the available niacin from food (Ball, 2005; Eitenmiller et
al., 2007).
adult women and men respectively (NHMRC, 2006). Deficiency of niacin will
cause pellagra, while a high dosage of niacin (3-6g/ day) can affect liver
Folate is the generic descriptor for folic acid and related compounds exhibiting
the biological activity of folic acid. The folates from natural sources usually have
a single carbon unit at N-5 and/or N-10 position. The single carbon units that
may be transported and stored by folates can vary in oxidation state from the
acid). Folate is sensitive to heat, oxygen, light, acid and alkali, but folic acid is
stable. Thereby, folic acid is always used in fortified foods and supplementation
absorption region of the folates are mostly 280 – 300 nm (Combs, 2012).
Animal organ meats like liver and kidney, avocado, nuts, whole grains, leafy
green vegetables and peas are all food sources of natural folate. The naturally
55
storage conditions, it will slowly degrade to monoglutamates and be oxidized to
less available folate (Ottaway, 1993). The bioavailability of folate in foods varies
from extrinsic factors such as chemical and physical form of the vitamin and the
concentration of the vitamin, food matrix and antagonists to the intrinsic factors
including animal species of the food and health of small intestine (Combs, 2008;
Eitenmiller et al., 2007; Zempleni et al., 2013). Folic acid has 85%
bioavailability, which is 50% more than other forms of folate as a result of its
from natural source found in foods such as THF and 5-MTHF has 75%
The recommend intake of folate is 400 μg/ day for adults, 600 μg/ day for
pregnancy and 500 μg/ day for lactation (NHMRC, 2006). Folate deficiency will
folate plays an important role in B12 deficiency, since B12 induced anemia can
be alleviated by treating with folic acid. If folate supplements are taken without
56
Free thiamin is unstable as its quaternary nitrogen will be cleaved to the thiol
form in water (Combs, 2012). For this reason, the hydrochloride and
however, they are unstable in alkaline solutions and are sensitive to ultraviolet
light and high temperature (Matarese & Gottschlich, 1998; Shils & Shike, 2006).
intake is high, only a small amount of the thiamin is absorbed and elevated
lean pork, macadamia nuts, sunflower seeds are all good sources for thiamin
(Combs, 2008). The recommended daily intake of thiamin is 1.2 mg/day for
adult men and 1.1 mg/day for adult women (NHMRC, 2006). The deficiency of
however, the excess of thiamin over 3 g/day will also lead to toxic headaches,
57
2.6.4 Vitamin B6
Vitamin B6 is a group of compounds, which can be divided into free form and
pyridoxic acid. Free form is mostly found in plants, while phosphorylated form is
are white to off-white platelets or rod crystals, and the commercial preparations
Good food sources of VB6 include meats, whole grain products, vegetables,
nuts and bananas. More than 50% of vitamin B6 may be lost during cooking,
Recommended daily intake of Vitamin B6 is 1.4 mg/day for adult people under
50; 1.7 mg/day for adult men over 50 and 1.5 mg/day for adult women over 50
58
dinucleotide that catalyze many oxidation-reduction reactions. The bioactive
forms of riboflavin are the oxidized and reduced forms of flavin adenine
dinucleotide (FAD and FADH2) and flavin mononucleotide (FMN and FMNH2)
(NHMRC, 2006).
the light intensity, exposure time, light wavelength, packaging materials and
Riboflavin is necessary for overall normal growth and development of the body,
production and regulation of certain hormones, and formation of red blood cells
and proteins and is crucial for the production of biological energy in the electron
transport system. It is also essential for the maintenance of healthy vision, skin,
depression.
skin and mucous membrane issues, such as chelosis, angular, stomatitis and
2013). The recommend daily intake (RDI) of riboflavin for adult men under 70 is
59
1.3 mg/day and 1.6 mg/day for men over 70; the RDI for adult women under 70
is 1.1 mg/day and 1.3 mg/ day for women over 70 (NHMRC, 2006). Liver, milk,
cheese, egg, some green vegetables and beer are all dietary sources of
riboflavin.
60
3. Materials and Methods
Fresh banana pseudostem from banana plants (Musa acuminata & Musa
balbisiana) was used for the experimental work. The banana pseudostem was
collected from the Royal Botanic Garden in Sydney. The banana pseudostem
was refrigerated immediately after collection and was dried within few days.
61
3.2 Sample Preparation and Drying
3.2.1 Equipment
The equipment for sample preparation and drying is listed in Table 3.1.
2. Drying trays
5. Sterile knife
6. Chopping board
7. Desiccator
8. Aluminum containers
The stored banana pseudo stem was taken out of the fridge and the outer layer
of the pseudo stem was peeled off leaving the core of the pseudo stem. The
tender core of the pseudo stem was cut transversally into 4 mm slices
62
using the slicer. Afterwards, the cut pseudo stem slices were separated into two
parts. Half the quantity was blanched at 70 °C water for 3 minutes and another
The drying tray used for drying was first weighed. Afterwards, blanched and
untreated samples were further divided into two parts separately. Samples were
spread out neatly onto the drying tray. Tray of banana pseudo stems was
weighed again before they were placed into a batch dryer. The temperature of
the dryer was set at 40±2 °C and 50±2 °C, respectively. The weight of banana
pseudo stem was recorded manually every hour until the weights were
constant. Banana pseudo stem samples were taken out of the dryer and the
final weight was recorded. The samples were placed immediately into a
desiccator to cool down. From these recorded weights, drying curves of the
samples were plotted on a graph with time in hours against weight loss in
M Me
MR (3-1)
Mo Me
63
3.3 Nutrient analysis
3.3.1 Moisture
3.3.1.1 Equipment
1. Aluminum containers
4.Desiccator
The initial moisture content of fresh and dried banana pseudo stem was
uses the air oven method. Triplicates of fresh and dried banana pseudo-stem
Then the samples in aluminum containers were dried in an air oven overnight at
105 ± 2 °C. After drying, the samples were taken out and placed into the
desiccators to cool down and final weights of the samples were recorded. The
moisture content of fresh and dried banana pseudo stem was determined.
The moisture contents (wet basis) were calculated using equation (3-2).
Wo- Wf
MCwb= 100 (3-2)
Wo
64
The moisture contents (dry basis) were calculated using equation (3-3).
Wo- Wf
MCdb= 100 (3-3)
Wf
3.3.2 Fat
3.3.2.1 Equipment
2. Beakers 50 mL
9. Glass beads
11. Spatula
65
3.3.2.2 Chemicals
The chemicals used for fat analysis are listed in Table 3.4.
Technology (NIST)
Total fat content in the banana pseudo stem samples was determined by acid
sample (2 g) were weighed into beakers. Blanks were run along with the
samples. Ethanol and 7 N HCl were added and heated for 40 minutes while
flask; 25 mL of diethyl ether were added and shaken vigorously for 1 minute.
Furthermore, 25 mL of petroleum ether was added into the flasks and inverted
gently without shaking. The upper ether-fat layer was decanted off and filtered
through a cotton pledget into pre-weighed Erlenmeyer flasks with glass beads.
The remaining liquid in the Mojonnier flasks were re-extracted twice with 15 mL
66
using both ethers followed by same procedure with decanting and filtered into
same flasks and the funnels were washed thoroughly with 10 mL of mixture of
the two ethers in equal volumes. Then, the ethers were slowly evaporated on a
steam bath in the fume cupboard. The Erlenmeyer flasks containing the fat
were dried in an oven at 100 ± 2 °C for 90 minutes; furthermore, the flasks were
allowed to stand in air for exactly 30 minutes and weighed to determine the final
fat content in the samples. A reference material obtained from NIST, US, typical
Wt of fat
%Fat in sample = 100 (3-4)
Wt of sample
3.3.3 Proteins
3.3.3.1 Equipment
Analyser
67
3.3.3.2 Protein analysis
Protein content of the samples was determined by using the LECO Analyser.
The percentage of nitrogen (N) in the samples was determined and the protein
conversion factor, 6.25 was used to convert the nitrogen content into protein
content.
3.3.4 Ash
3.3.4.1 Equipment
Crucibles
Aluminium containers
Desiccator
Spatula
68
The ash content of banana pseudostem was quantified by dry ashing method
942.05 (AOAC, 2006). The crucibles were prepared by washing with warm
soapy water and rinsing in tap water. The crucibles were soaked in 7.5 M HNO3
overnight in a fume cupboard. The crucibles were then rinsed with deionized
water and dried in an air oven at 110±2 °C, followed by conditioning in a muffle
furnace at 530±2 °C overnight. Finally the crucible was cooled and stored in a
The previously conditioned crucibles with lid were weighed. Duplicate of each
sample (4 g) were weighed into the crucibles. Blanks were run along with the
samples. Then the samples were charred on a hot plate. The crucible and lid
were placed into the muffle furnace at 530 ± 2 °C overnight. If the ash was still
grey, a few drops of 7.5 M Nitric acid was added to the cooled crucible and
evaporated to dry on a hot plate in the fume cupboard and returned to the
taken out and placed into a desiccator to cool and then reweighed in order to
(3-6)
69
3.3.5 Sugars
3.3.5.1 Equipment
Beakers 100 mL
Measuring cylinder 25 mL
pH meter TPS
Syringe 10 mL
70
3.3.5.2 Chemicals
The Chemicals used for sugar analysis are listed in Table 3.8.
Sugars were extracted with aqueous ethanol following the AOAC method
were weighed into each beaker. Whatman No.541 filter papers were also pre-
measured. Then, a few drops of 0.5 NaOH were added to increase the pH to
containing food samples and placed it in a steam bath for a few minutes,
covered with a watch glass and stirred frequently. The solution was then
removed from the steam bath and filtered through the filter paper into a round
bottom flask. The extraction was repeated three times with 25 mL of boiling
85% ethanol. Then the flask containing the ethanol solution was evaporated off
71
approximately 3 mL. The aqueous solution was transferred into a 10 mL
volumetric flask and made up to volume with milliQ water. Furthermore, the
3.3.6 Starch
3.3.6.1 Equipment
PH meter TPS
Volumetric flasks 1L
Autosampler vials
Beakers 100 mL
Syringe 10 mL
72
3.3.6.2 Chemicals
The chemicals used for starch analysis are listed in Table 3.10.
Amylogluosidase Megazyme
Invertase Sigma-Aldrich
Pancreatin Sigma-Aldrich
Pullulanase Novozymes
The residues from sugar extraction were further used for starch extraction. 350
73
mg residue of each sample was dissolved into 10 mL Milli-Q water. The
dissolved samples were sealed with foil and placed into a boiling shaking water
bath for 4 hours. 0.3 mL acetate buffer was added, followed by addition of 0.4
were filtered through Whatman No. 541 filter paper. Aliquots of 25 mL were
solution was left. Then the solution was passed through a 10 mL syringe and
Sugar and starch content was determined by using HPLC with Refractive Index
column was used in the analysis. Acetonitrile and MilliQ water (87:13) was used
as the mobile phase. The conditions were: 20 μL injection, isocratic mode, flow
rate of 0.4 mL/min and temperature control of column set to 30 °C. Samples
and standards were injected to determine the final weight of sugar and starch in
methods described by Englyst et al. (1992) with slight modifications. The recipe
for glucose measurement was modified. 100 μL samples mixed with 2 mL GOD-
74
GOD-PAP
Enzyme solution 1
g pancreatin was weighed into each of the four centrifuge tubes and each
magnetically for 10 min and then centrifuged for 10 min at 1500g. 13.5 mL
supernatant from each tube was pipetted out into another tube and mixed with 6
acid solution, and made upto 1 L with water. pH was adjusted to 5.2 with 0.1 M
acetic acid. To stabilize and activate enzymes, 4 mL 1 M CaCl2 was added into
a liter of buffer.
200 mg glucose (dried to constant weight) was weighed into a test tube to the
nearest 0.1 mg. The volume was made up to 20 mL with sodium acetate buffer
to give a 10 mg/mL solution. The following gradient table was used to build the
standard curve.
75
Table 3.11 Standard concentration table
0.1 2 18
0.5 1 19
Dried ground banana pseudostem (500 mg) was mixed with 0.1 M acetate
buffer and 51.5 cm diameter glass balls in a test tube and vortex mixed
vigorously. The tubes were placed into a boiling water bath for 30 min. Tubes
again. When the temperature stablised, 0.2 mL invertase was added and the
tubes were capped and immersed horizontally in the shaking water bath at 37
°C for 30 min.
The tubes were taken out of the water bath and manually shaken vigorously.
0.2 mL of the contents was removed into eppendorf tubes containing 1mL
absolute ethanol and vortex mixed. They were then centrifuged at 1,500 × G for
inversion.
76
3.3.6.5.3 Measurement of RDS, SDS and RS fractions
Guar gum powder (50 mg) was added and 20 mL of 0.1 M acetate buffer was
pipetted into each sample tube with 500 mg dried ground banana pseudostem
and 51.5 cm diameter glass balls. The contents were mixed thoroughly to
disperse the guar gum. The samples, standards and blank were placed in a
water bath at 37 °C. Into each tube was added 5 mL enzyme solution 1. After 1
min the second sample was added. The tubes were capped and immersed
After 20 min, 0.2 mL of the hydrolysate was pipetted out into a labeled
eppendorf tube containing 1 mL absolute ethanol and mixed well. The tube was
placed back into the shaking water bath. After a further 100 min, a second 0.2
mL sample was removed from the same tube in the same way, but this time the
tube was not placed back into the water bath. The portion taken after 20 min
The samples were vortex mixed after 120 min to break up large particles. The
tubes were placed in a boiling water bath for 30 min. The tubes were then
vortex mixed again and cooled in ice water for 15 min.10 mL of 7M KOH was
added and mixed well. The tubes were immersed horizontally in the ice-water
shaking bath (0 °C) for 30 min. Removed the sample tubes from the ice water
and 0.2 mL of the contents were pipetted into eppendorf tubes containing 1 mL
0.5 M acetic acid. The contents were mixed well. 0.2 mL diluted
amyloglocosidase (50 AGU/ml) was added, mixed and incubated for 30 min at
77
70 °C. The tubes were transferred into a boiling water bath for 10 min and
cooled to room temperature. 0.2 mL sample was pipetted and diluted with 1 mL
water. The tubes were centrifuged at 1,500 × G for 5 min to remove the
precipitate.
10 μL blank, samples and standards in duplicate were pipetted into labeled test
tubes. 270 μL GOD-PAP reagent was added into each and mixed. Each mixture
was pipetted into the microplate and incubated at 37 °C for 20 min. The
RS = TS – (RDS + SDS)
78
3.3.7 Dietary fibre
3.3.7.1 Equipment
The equipment used for dietary fibre analysis is listed in Table 3.12.
Fritted crucible
Heavy-walled filtering 1L
flask
Desiccator
PH meter TPS
Cylinder 1L
stirring bars
Spatulas
79
Vacuum pump
NMR
Agilent 7890A
80
3.3.7.2 Chemicals
The chemicals used for dietary fibre analysis are listed in Table 3.13.
4. Protease Megazyme
5. Amyloglucosidase Megazyme
--aminomethane (TRIS)
81
Table 3.13 Chemicals used in dietary fibre analysis (Cont’d)
Total dietary fibre, which consists of soluble and insoluble dietary fibre in the
suitable for cereal products, fruits and vegetables (Megazyme Total Dietary
82
fibre assay procedure, 2012). The method was based on AOAC method 991.43
Duplicates of each sample (1 g) were weighed into beakers. Blanks were run
added to each beaker. The beakers were placed on the magnetic stirrer with the
addition of a magnetic stirring bar until all samples were completely dispersed in
solution. Then the samples were incubated in order with three different
covered with foil and placed into the shaking water at 100 °C to incubate for 35
minutes. After incubation, the beakers were cooled to 60 °C and the sidewalls of
the beakers were rinsed with distilled water. The second incubation was
initiated after addition of 100 μL of protease solution. This time the incubation
temperature was set to 60±1 °C and the time was 30 minutes. After second
incubation, 5 mL of 0.561 N HCL was added into the beakers while stirring.
into the beakers and incubated in the shaking water bath at 60±1 °C again for
30 minutes. The samples were removed from the water bath after incubation
and filtered. The crucibles containing Celite® was weighed and the Celite® was
mixture from above was filtered through the crucible into the filter flask using
vacuum. The residue in the crucible was washed twice using 20 mL of distilled
83
Insoluble dietary fibre
The filtrates were saved for determination of soluble dietary fibre. The residue in
the crucibles was washed twice with 10 mL of 95% ethanol and acetone. Finally
The filtrates from above were transferred into pre-weighed beakers. Then 4
volumes of 95% ethanol preheated to 60 °C were added into the beakers. The
beakers were left at room temperature for 60 minutes to allow for the formation
of precipitate. After that, new crucibles containing Celite® were weighed and the
Celite® was redistributed using 15 mL of 78% ethanol and vacuum source. The
precipitated enzyme digest was filtered through the crucible with vacuum and
the residue in the crucibles was washed with 78% ethanol, followed by 95%
ethanol and finally acetone. Again, the crucibles were dried in the air oven at
103 °C overnight.
On the next day the crucibles were removed from the oven and cooled in the
desiccators for approximately 1 hour. The crucibles were weighed and the
weight of residue was obtained by subtraction. The residue from one duplicate
sample was analysed for protein and then the second duplicate sample was
analysed for ash. For protein, the residues were determined by Kjeldahl
method. For ash, the crucibles containing residue were placed into the muffle
furnace for 5 hours at 525 °C. Then they were taken out and cooled in the
84
desiccators and the weight was recorded
R1 R 2
p AB
Dietary fibre (%)= 2 100 (3-7)
m1 m 2
2
where:
BR1 BR 2
B BP BA (3-8)
2
Soluble and insoluble dietary fibre extraction methods were followed by the
Megazyme method described above. The dried residues of both soluble and
13
C CP/MAS NMR spectra of samples were measured using Bruker Avance III
85
4 kHz).
Dissolved 8.2 g of sodium acetate and made to 1 L with MillQ water. Adjusted to
pH 5.2 with 0.1 mol/L acetate acid. In order to stabilize and activate the
beaker in a bowl of ice water in a fume cupboard and slowly added 390 mL of
Enzyme solution 1
Took 2.5 mL of Termamyl and made up to 200 mL with acetate buffer, mixed
Enzyme solution 2
mixed initially and then mixed for 10 min with a magnetic stirrer. Vortex mixed
86
added 2.5 mL of pullulanase and vortex mixed again.
determined using the methods described by Englyst et al. (1994) with minor
modifications.
A sample aliquot of 300 mg sample was weighted into the glass tube with 2 mL
boiling water bath and were removed after 20 s, vortex mixed and immediately
placed into the boiling water bath. This procedure was repeated until all the
After 30 min, one tube was removed at a time, vortex mixed, uncapped and 8
mL of enzyme solution 1 (keep at 50 °C) was immediately added, the tube was
capped and vortex mixed thoroughly, ensuring that no material adhered to the
tube wall. Tubes were replaced in the boiling water bath for 10 min. The rack of
tubes was transferred to a 50 °C water bath. After 3 min, the rack was removed
and 0.5 mL of enzyme solution 2 was added to each tube and the contents
were mixed thoroughly to aid distribution of the enzyme throughout the sample.
The tubes were replaced in the 50 °C shaking water bath and left for 30 min.
The rack of tubes was transferred to the boiling water and left for 10 min. The
For total NSP, 40 mL of absolute ethanol was added to each sample and they
were mixed well by inversion. They were then placed in ice water for 30 min.
87
supernatant liquid. Removed as much of the supernatant as possible by
pipette.10 mL of 85% ethanol was added to the samples and vortex mixed.
For insoluble NSP, added 40 mL of sodium phosphate buffer and put in the
boiling shaking water bath for 30 min. After cooling down the samples,
was added to the samples and vortexed. Then made to 50 mL with water,
absolute ethanol.
For both treated total NSP and insoluble NSP, uncapped tube was placid in a
beaker of water at 80 °C on a hot plate stirrer in the fume cupboard and the
residue was stirred until dry to make sure all of the samples were free of
acetone.
vortex mixed. Left the tubes in shaking water bath at 35 °C for 1 h to disperse
the cellulose. Added 25 mL of water rapidly and vortex mixed. Placed into a
shaking boiling water bath and left for 1 h. Cooled the tubes in tap water at
room temperature.
The acid hydrolysed samples from 3.3.7.5 were neutralized with 1.5 g of
88
supernatant liquid using centrivap and re-dilute with 1 mL D2O (100%).
1
H NMR spectra of samples were measured using Bruker Avance III HD 600
obtained by pulse program zgesgp with 90 degrees pulse. The solvent used
For the separation of neutral sugars, a Supelco SP-2380 wide bore capillary
(Agilent, 7890A, USA) and auto-injector (Agilent, 7683B, USA) was used. The
injector temperature and detector temperature were set to 275 °C and the
column temperature was set to 220 °C. Helium was selected as the carries gas
and the flow rate was 8 mL/min. A split/ splitless topered liner (Agilent, USA)
was used for the injection. The injection volume was 1 L and a split injector
89
with a dilution factor of 30 was used.
Standard stock solution and internal standard (allose) stock solution were
separately prepared in 50% saturated benzoic acid. Made the internal standard
pressure with phosphorus pentoxide) to the nearest 1 mg. Diluted to 100 ml with
50% saturated benzoic acid to give a 1 mg/mL solution. The stock sugar
solution was made by weighing (all sugars dried to constant mass under
calibrated flask and diluted to volume with 50% saturated benzoic acid. All of
the stock solutions were stored in a refrigerator covered with aluminum foil.
50% saturated sugar mixture, which was used to build up the standard curves
Table 3.14.
90
Table 3.14 Calibration concentration of the standard sugar mixture (mg/ L)
Rha: Rhamnose; Fuc: Fucose; Ara: Arabinose, Xyl: Xylose; Man: mannose;
The method described has been validated with respect to accuracy, within-day
examined neutral sugars, covering the entire working range; nine concentration
levels were used. Each solution was injected twice. Correlation coefficients for
/internal standard sugar) against peak area ratio (external standard sugars/
91
The detection limits and quantitation limits were considered to be the quantities
that produce a signal of peak height three times the size of the background
noise.
The sample preparation for the GC analysis followed the method described by
Englyst et al. (1994). To prepare the standard sugar mixture, 0.2 mL of the
sugar mixture solution and 0.5 mL of 2.4 mol/ L sulfuric acid were mixed.
Duplicate standard sugar mixtures at each concentration level were used for
calibration.
The tubes were placed in ice-water and 0.4 mL of 12 mol/L ammonia solution
tetrahydroborate solution were added and vortexed. The sample tubes were left
in a 40 °C water bath for 30 min. 0.2 mL of glacial acetic acid was added and
mixed again. 0.5 mL sample was transferred to a 50 mL glass tube and add 0.5
immediately. 0.9 mL of absolute ethanol was added. After being vortex mixed,
samples were left 5 min for reaction. 10 mL water was added, vortex mixed and
left for 5 min. 0.5 mL Bromophenol Blue solution was added. The tubes were
placed in ice water and 7.5 mol/ L potassium hydroxide was added; 5 min later,
92
a further 5 mL of 7.5 mol/L potassium hydroxide was added. The samples were
mixed by inversion. The samples were centrifuged at 5000 × G for 5 min. Part of
3.3.8 Minerals
3.3.8.1 Equipment
Ion Chromatography
3.3.8.2 Chemicals
The chemicals used for mineral analysis are listed in Table 3.16.
93
3.3.8.3 Procedure of mineral analysis
with nitric acid and hydrogen peroxide before being analysed by the ICP-OES.
94
3.3.9 B-complex Vitamins
3.3.9.1 Equipment
The equipment used for B-complex vitamins analysis is listed in Table 3.17.
1. Beakers 100 mL
3. pH meter TPS
visible diode-array
detector (PDA)
Column
cartridge
10. Syringe 10 mL
95
3.3.9.2 Chemicals
The chemicals used for B-complex vitamins analysis are listed in Table 3.18.
Biphenyl 100A (150*4.6 mm, 2.6 m) was used at ambient temperature. The
96
modification. The mobile phase of the HPLC system was delivered at a flow rate
B v/v composition of 99:1, and remaining isocratic for 8 min. This composition
was changed linearly to reach 40% of solvent B after 18 min, to reach 55% of
solvent B after 28 min, reach 100% of solvent B after 33 min and finally elution
Detection was performed with a photodiode array detector (PDA) at 280 nm.
follows: 260 for nicotinic acid, 257nm for pyridoxine, 256 nm for nicotinamide,
270 nm for thiamin, 280 nm for folic acid, 290 nm for 5-methylfolate and 265 nm
executed by comparing their spectra with those derived from standard solutions.
The aqueous stock and standard solutions of the B-complex vitamins were
of the stock solutions was 250 ng/ L, while the concentration of the working
solutions ranges from 0.25 ng/ L to 50 ng/ L. The stock solutions were stored
in a refrigerator and covered with aluminum foil in order to protect them from
97
3.3.9.3.3 Method validation
The method described has been validated with respect to accuracy, within-day
concentration levels were used in the range 0.25-50 ng/ L. Each solution was
injected three times. Correlation coefficients for the B-complex vitamins on the
basis of plots of concentration (ng/ L) against peak area (maul) were found to
be >0.996.
The extraction procedure used in this study was based on a combination of acid
of standard solution was mixed with 8 mL 0.1 N hydrochloric acid and boiled in
the water bath for 20 min. After cooling down the sample, the pH of sample was
correct for the addition of vitamins from the enzyme suspension. After 18 h
inactivate the enzyme. After cooling down, the samples were centrifuged for 10
98
min at 14,000 × G. The supernatant was kept for SPE.
(55 m, 70A) cartridge was used for purification. The SPE method of Cho et al.
(2000) was used of the extraction of water soluble vitamins. The stationary
phase was flushed with 10 mL methanol and 10 mL acidified Milli Q water (pH
4.2) to activate the stationary phase. The supernatant of the sample was loaded
and eluted with 5 mL Milli Q water (pH 4.2) then 10 mL methanol at a flow rate
of 1 mL/ min. The eluent was collected in a bottle covered with foil and
solution was passed through a 10 mL syringe and filtered with PTFE membrane
99
3.4 Physicochemical properties analysis
3.4.1.1 Equipment
Magnetic stirrers
Stirring bars
Centrifuge tubes 50 mL
The water holding capacity was determined based on the standard methods
weighted centrifuge tube. It was stirred with a magnetic stir bar at room
temperature for 30 min and left for 30 min. It was then centrifuged at 3500 rpm
for 30 min.
100
After that, the supernatants were decanted, each centrifuge tube was weighed
then the WHC was calculated as g water of dry sample using equation (3-9).
WHC
wt of centrifuge tube after decanting wt of dry tube total sample wt
total sample wt (3-9)
solubility analysis
3.4.2.1 Equipment
Magnetic stirrers
Stirring bars
Centrifuge tubes 50 mL
The determination of SWC and solubility were following the method suggested
accurately weighed and quantitatively transferred into a clear dried test tube
101
distilled water using a stirrer. The resultant slurry was heated at the 95 °C for 30
Aliquots (5 mL) of the supernatant were dried to a constant weight at 110 °C.
The residue obtained after drying the supernatant represented the amount of
The residue obtained from the above experiment (after centrifugation) with the
water it retained was quantitatively transferred to the clean dried test tube used
3.4.3.1 Equipment
(XRD). The X’Pert PRO Multi-purpose X-ray Diffraction System (MPD system)
was used in this experiment. The banana pseudostem powder was packed in a
102
standard sample holder with 30 mm diameter and 2.5 mm depth. After packing,
all of the samples were placed on the automatic multi-sample changer for X-ray
diffraction. The XRD was operating at tension of 45 kV/ 40 mA. Scans were
obtained from 4 to 55 degrees 2θ in 0.026 degree steps for 50.05 s per step.
extracted by a curve fitting process from the diffraction intensity profiles. A peak
functions for each peak and a broad peak at around 21.5 °C assigned to the
from the diffractogram of the whole sample, the CI was calculated by dividing
the remaining diffractogram area due to crystallinity by the total area of the
original diffractogram.
103
4. Drying of banana pseudostem
banana pseudostem
The drying curves show variations of the moisture ratio versus drying time for
shown in Figure 4.1. The moisture ratio is defined as the ratio of moisture
content of the sample at each moment (M) and initial moisture content (Mo) of
the sample (Agarry et al., 2013). The patterns of the curves obtained under 4
shapes.
At the initial stages of the drying process, the drying rates were high and with
104
the increase in time the drying rates decreased until the samples reached the
conditions (temperature and relative humidity) the end of the drying process
was attained. The reason for this trend to occur is due to the migration of water
out of the sample (Fellows, 2002). Initially, the surface of the samples heated up
to wet bulb temperature and the free water moved out of the samples and
moved from the interior of the material to the surface following a falling rate of
drying. Figure 4.1 shows that drying at 50 °C without pretreatment exhibited the
fastest drying rate. This drying temperature appears to be suitable for the
industry as it can save both energy and cost. This is due to the fact that an
increase in air temperature causes water to evaporate more rapidly from a wet
balbisiana pseudostem, and this condition was also selected to dry Musa
acuminata pseudostem.
105
4.1.2 Effects of species on the drying curve
Figure 4.2 shows the drying curves of these two different species.
According to the figure above, Musa balbisiana pseudostem has a higher drying
rate than Musa acuminata pseudostem, which means the loss of moisture from
Musa balbisiana pseudostem per each unit of time is higher than that from
dry, while Musa acuminata pseudostem requires 6 to 7 hours to dry. This may
indicate that there is less free water and more bound water in Musa acuminata
Table 4.1 shows the colour (L*, a*, b*) of BP40, BP40B, BP50, BP50B and
106
BPA. L* value reflects the lightness of the samples, the higher L* value is, the
whiter the sample is. The a* values are indicative of the red and green colours,
indicates green colour. The b* values are indicative of yellow and blue colour, in
which a negative b* value indicates blue and a positive b* value indicates yellow
L* a* b*
different (P<0.05).
Significant difference (p<0.05) of colour (L*, a* and b*) was observed in Musa
highest L* value and lowest b* value, while BP40 showed the lowest L* value
and highest b* value. This result indicates that the banana pseudostem dried at
colour. It is believed that blanching can inactivate enzyme and inhibit colour
107
deterioration. Blanching was proved to progressively decrease
peeled bananas (Castro et al., 2008; Cano et al., 1990). However, blanching
samples in this study didn’t provide a whiter colour than samples without
blanching. One possibility is that the samples collected in this study were fresh.
There was several hours gap between collection and sample preparation. The
oxidation was still at the beginning stage and there was not much browning
effect on the samples. Despite the fact that BP50 provided the whitest colour,
Figure 4.1 also shows that BP50 had the highest drying rate, while BP40 had
the lowest drying rate. Thus, in the case of drying and white colour preservation,
provided higher L* and lower b* value than Musa acuminata pseudostem, which
indicated that Musa balbisiana pseudostems were whiter than those of Musa
difference in colour between the two banana species under the sample
Water activity differs from moisture content. Water activity measures the
availability of free water in a food system that is responsible for any biochemical
reactions, while the moisture content represents the water composition in a food
system (Fazaeli et al., 2012). Water activity of dried food below 0.6 is
108
believed to be good for food preservation as no microbes can multiply at this
pseudostem in this study were below 0.6. This means all of the drying methods
Table 4.2 Water activity (aw) of Musa balbisiana and Musa acuminata
aw
BP40 0.41
BP40B 0.51
BP50 0.40
BP50B 0.35
BPA 0.41
109
5. Effects of drying on the nutrient stability of banana
pseudostem
Table 5.1 Mean proximate contents of Musa balbisiana and Musa acuminata
Musa balbisiana
Musa acuminata
different (P<0.05).
All values are based on dry weight basis and means of replicate determinations
5.1.1 Moisture
BP40, BP40B, BP50, BP50B were 6.2%, 5.0%, 4.4% and 4.5% respectively
110
(Table 5.1). These values were lower than those reported in literature that
oven drying method. Both Aziz et al. (2011) and Bhaskar et al. (2011) reported
higher moisture contents in dried banana pseudostem (8.82% and 15.1%). The
time). Thicker pieces of cut banana pseudostem would take longer time to dry
and the moisture content in these dried pieces would be higher than in thinner
dried pieces. The banana species used in the studies of Aziz et al. (2011) and
Bhaskar et al. (2011) were different from this study, which may cause the
difference in the moisture content. The banana species collected from Bhaskar
et al. (2011) is Musa sp. var. elakki bale and from Aziz et al. (2011) was the
50 °C had the lowest moisture content, while samples dried at 40 °C had the
highest moisture content. This result was similar to the drying curve obtained at
50 °C without pretreatment which was the optimal condition for drying banana
(P>0.05), but there was significant difference between BP40, BP50 and BP40B
or BP50B. This result implies that temperature has no effect on the drying
compared with 4.4% of dried Musa balbisiana pseudostem. These two values
were significantly different (P<0.05). This result correlates with the result
111
that drying at 50 °C without pretreatment has the highest drying rate. This may
pseudostem.
5.1.2 Ash
As shown in Table 5.1, the ash content found in BP40, BP40B, BP50, BP50B
and BPA samples was 12.9%, 12.3%, 14.0%, 13.4% and 15.9%, respectively.
All values reported in this study were on a dry weight basis. Significant increase
in ash content was observed with the increase in drying temperature (P < 0.5).
No significant change in ash content was found between BP40 and BP40B (P >
0.5), but the ash contents of BP50 and BP50B were significantly (P<0.05)
different. These results indicate that drying temperature has an effect on ash
results, the value of ash varied with different drying temperatures and banana
species.
Ho. et al. (2012) and Aziz et al. (2011) reported lower ash contents, which were
6.75% and 10.08%. Higher ash content in this study indicates higher mineral
this study was the tender core. It is likely that minerals are concentrated in the
112
tender core of the pseudostem. Another possibility is that the banana species
5.1.3 Protein
The protein content of BP40, BP40B, BP50 and BP50B were 3.1%, 3.2%, 3.4%
and 3.4%. All values are based on a dry weight basis. There is no significant
difference (P>0.05) among these values. Temperature and blanching did not
affect the protein values in Musa balbisiana pseudostem. Bhaskar et al. (2011)
reported lower protein content in banana pseudostem, which was 2.5%. The
differences in the values may be due to the various methods used to analyze
protein. Aziz et al. (2011) showed similar protein in tender core of banana
pseudostem flour with this study, which was 3.52%, but the protein content of
the native banana pseudo-stem flour was only 0.89 in their study. The protein
than Musa balbisiana pseudostem (3.4%), which was nearly double the value of
(NUTTAB, 2010), which was 10.8 g/100 g, the protein contents in both banana
pseudostem species are low. It implies that dried banana pseudostem flour has
5.1.4 Fat
The fat contents of BP40, BP40B, BP50, BP50B and BPA were found to be
3.2%, 4.0%, 3.4%, 3.0% and 2.8%. There was no significant difference (P>0.05)
113
Musa acuminata pseudostem was slightly lower than in Musa balbisiana
pseudostem, but there was no significant difference between these two species.
These values were higher than those reported by others for dried banana
pseudostem, which had a fat content of 1.18% and 1.7% (Aziz et al., 2011 &
5.1.5 Carbohydrates
The total carbohydrates in BP40, BP40B, BP50, BP50B and BPA were 66.3%,
66.2%, 64.4%, 68% and 62.7%. Although BP50B had the highest carbohydrate
content, there was no significant difference (P >0.5) between all the banana
5.2 Micronutrients
5.2.1 Minerals
The mineral contents in the two species are reported in Table 5.2. The mineral
copper, iron, zinc, manganese and selenium. The predominant elements found
For Musa balbisiana, potassium was the most predominant (5374 mg/100 g,
5015 mg/100 g, 6091 mg/100 g and 5524 mg/100 g for BP40, BP40B, BP50,
114
BP50B, respectively) followed by Mg (197 mg /100 g, 200 mg/100 g, 255
mg/100 g and 233 mg/100 g for BP40, BP40B, BP50, BP50B, respectively), Ca
(115 mg/100 g, 130 mg/100 g, 129 mg/100 g and 145 mg/100 g for BP40,
mg/100 g and 64 mg/100 g for BP40, BP40B, BP50, BP50B, respectively). For
acuminata was 262 mg/100 g, which was almost double that of the Musa
balbisiana.
Table 5.2 Mean mineral content (mg/100 g) of Musa balbisiana and Musa acuminata
Ca Cu Fe K Mg Mn Na P Se Zn
Musa balbisiana
40 115 0.5 8.5 5374 197 0.7 0.7 59 0.1 16.6
40 B 130 0.6 6.8 5015 200 0.5 13.2 65 0.1 11.1
50 129 0.4 9.4 6091 255 0.7 1.5 69 0.1 22.4
50 B 145 0.9 11.6 5524 234 0.7 13.6 64 0.1 13.8
Musa acuminata
BPA 262 0.5 8.5 6963 230.4 0.9 1.4 163 0.0 9.5
All values are based on a dry weight basis. All values are means of replicate
determinations
However, the results obtained from the present study differed from those
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pseudostem (Musa acuminata), which was the same species as in this study,
content was almost 6 times that found in the present study. Meanwhile, the
potassium content reported by Ho et al. (2012) was 944.12 mg/ 100 g, which
was only about one sixth of that in the present study. The reason for this may
collected. Ca concentration increased with age (Ho et al., 2012). More mature
the present study. Therefore the calcium content in the study of Ho et al. (2012)
was significantly higher and the potassium was lower than in this study.
the pseudostem decreases during the fruiting phase, because the bananas
require substantial amounts for fruit development (Ho et al., 2012). At the
other cations, especially potassium (Ho et al., 2012). The potassium in the
banana pseudostem found in this study was extremely high; it even exceeded
the amount of banana fruit, which is considered as a high potassium food. The
K content of raw peeled Cavendish banana and ladyfinger banana are 346 and
322 mg/100 g (NUTTAB, 2010, wet weight basis), whereas the K of Musa
balbisiana and Musa acuminata pseudostem in this study were both over 450
mg/ 100 g (wet weight basis). Banana pseudostem could be a good source of
mg/100 g and 13.6 mg/100 g in BP40B and BP50B compared with 0.7 mg/100
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g, 1.5 mg/100 g and 1.4 mg/100 g in BP40, BP50 and BPA. This may be
because the banana pseudostem was in contact with water when blanched and
In the case of micro-elements, the contents of Cu, Fe, Zn, Mn, and Se were
found to be low in this study. Among these elements, Zn was the highest, which
was 16.6 mg/100 g, 11.1 mg/100 g, 22.4 mg/100 g, 13.8 mg/100 g and 9.5
mg/100 g in BP40, BP40B, BP50, BP50B and BPA, respectively. Iron was the
next highest with 8.5 mg/100 g, 6.8 mg/100 g, 9.4 mg/100 g and 11.6 mg/100 g,
8.51 mg/100 g in BP40, BP40B, BP50, BP50B and BPA, respectively. These
values were higher than those described by Ho et al. (2012), which were 8.05
mg/100 g for Zn and 3.31 mg/100 g for Fe. The difference may be due to the
balbisiana and Musa acuminata, while the pseudostems used in the study of Ho
et al. was the hybrid of Musa acuminata and Musa balbisiana, Compared with
other dried fruits and vegetables, such as dried apple, apricot and tomato, the
The normal level of Mn in plants ranged from 2.0-50.0 mg/100 g (Ho et al.,
2012), whereas this study indicated that the Mn contents in banana pseudostem
were lower than the normal range, which were 0.7 mg/100 g, 0.5 mg/100 g, 0.7
mg/100 g, 0.7 mg/100 g and 0.88 mg/100 g in BP40, BP40B, BP50, BP50B and
to four drying treatments were high in minerals, which agree with the high ash
117
described in chapter 5.1.2. The total amount of minerals of Musa balbisiana
detected in this study was 6579 mg/ 100 g and of Musa acuminata pseudostem
was 7639 mg/ 100 g, which were nearly half the values of ash contents.
5.2.2.1 Chromatography
There are two operation modes for HPLC separations, including isocratic and
gradient elution is most frequently used in chemical separation, which elutes the
The method used in this study was followed using the method of
Chatzimichalakis et al. (2004) with modification. The gradient elution using the
H2O/CH3OH (50/50) was selected for the separation of seven water soluble
99:1, and remaining isocratic for 8 min. This composition was changed linearly
to reach 40% of solvent B after 18 min, to reach 55% of solvent B after 28 min,
reached 100% of solvent B after 33 min and finally elution was performed
injections. The pH of the mobile phase is an extremely critical factor for the
118
impact the chromatographic selectivity, peak shape and retention (Dabre et al.,
2011). Without pH adjustment all the vitamins were separated except thiamin
and 5-MTHF (Figure 5.1a). Therefore, the influence of the mobile phase pH
within the range from 3.5 to 7.5 was studied, and it was found that thiamin and
m AU(x100)
9.0 260nm 1nm (1.00)
8.5 a
8.0
5-MTHF
7.5
7.0
6.5
6.0
5.5
5.0
Thiamin
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
-0.5
18.0 19.0 20.0 21.0 22.0 23.0 24.0 25.0 26.0 27.0 m in
m AU(x1 00)
6.0 26 0n m ,1 nm (1 .00)
5.5
HF
5.0
-M
5 T
b
4.5
m e
in
4.0
h
Tia
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
-0.5
20 .0 22 .5 25 .0 27 .5 30 .0 m in
Figure 5.1 Chromatography of standard thiamin and 5-MTHF with pH adjustment (a) and
119
shown in Figure 5.2. Retention times of examined vitamins were: 1. Nicotinic
p
mAU
Riboflavin
350
325
300
275
250
225
200
175
Nicotinic Acid
5-MTHF
150
125
Folic acid
Thiamine
100
Nicotinamide
75
50
Pyridoxine
25
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0 42.5 45.0 47.5 50.0 min
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5.2.2.2 Method validation
Table 5.3 Linearity of standard curves and sensitivity for the seven B-complex vitamin.
The calibration graphs were constructed by plotting the peak area based on
coefficient ranged from 0.9965 to 0.9997. The limit of detection (LOD) of the
noise of the standard finally diluted in the same buffer as the banana
thiamin, folic acid, 5-methyfolate and riboflavin were 0.06, 0.17, 0.08, 0.11,
0.05, 0.16 and 0.04 ng/ L, respectively. The limit of quantification (LOQ) was
calculated as ten times SD. The values for nicotinic acid, pyridoxine,
nicotinamide, thiamin, folic acid, 5-methylfolate and riboflavin were 0.20, 0.58,
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0.27, 0.38, 0.17, 0.54 and 0.13 ng/ L. LOD and LOQ proved to be comparable
improve the quality of vitamin analysis in food (Ollilainen et al., 2001). The
accuracy of the method was estimated by analyzing certified material BCR 485
(mixed vegetable). The BCR reference material can be used to verify the
accuracy of results and to monitor the performance of the method. The values
expressed as mean, standard deviation and recovery are reported in Table 5.4.
Table 5.4 Result of HPLC analysis of B group vitamin concentrations in certified reference
(SD).
The analytical results obtained for the reference material were comparable to
the certified values and the recoveries ranged between 91.9% and 106.3%.
Moreover, the banana pseudostem samples were spiked with known amounts
of the method.
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The intra-day repeatability was evaluated based on 10 measurements of the
same sample, independently prepared, within the sample set. The inter-day
consequent days. Validation results obtained for the vitamins analyzed are
The RSD for the same day ranged between 0.6% and 2.5% for peak areas. The
RSD between days were slightly higher (4.2%-9.7%) than the intra-day RSD
values. However, both of the RSDs of intra-day and inter-day experiments were
below 10%. The recovery rate of the vitamins in the intra-day experiment
ranged between 92.54% and 105.25%, while the recovery rate in the inter-day
123
Table 5.5 Inter-day (over a period of 7 consecutive days) and intra-day (n=6) values for the B-
complex vitamins
Intra‐day (n=10) Inter‐day (n=7)
124
determination of the seven B group vitamins in the banana pseudostem. The
peaks were identified by the following two steps. Firstly, by comparing the
retention time obtained from the samples, standards and the samples spiked
with the standards under identical conditions. Secondly, by comparing the UV-
in the banana pseudostem samples. The data obtained are given in Table 5.6.
125
Table 5.6 The concentration of B-complex vitamins (μg/g) in banana pseudostem dried in
Different species
Musa balbasiana
Musa acuminata
deviations (SD).
Nicotinic acid, pyridoxine and thiamin were found in banana pseudostem, while
nicotinamide, folic acid, 5-methylfolate and riboflavin were not detected in the
shown in Table 5.6, the concentration ranges for nicotinic acid, pyridoxine and
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thiamin of Musa balbisiana dried in different conditions were 0.24-0.8, 6.72-
10.11 and 2.05-2.23 μg/g respectively. Compared with dried fig (0.05 mg/100
g), currant (0.11 mg/100 g), litchi (0.01 mg/100 g), banana (0.18 mg/100 g) and
mixed fruit (0.10 mg/100 g), banana pseudostem (0.205- 0.223 mg/ 100g) had
higher thiamin content (NUTTAB, 2010 & USDA, 2007). Dried banana
dried apricot (0.1 mg/100 g), plums (0.21 mg/100 g) and litchi (0.09 mg/100 g)
and similar value with dried prunes (0.75 mg/100 g; NUTTAB, 2010 & USDA,
associated with the high level of pyridoxine in banana fruit. There was 0.2 mg
values of other raw fruit reported in NUTTAB Database (2010) were all below
0.1 mg/100 g. The niacin content detected in banana pseudostem powder was
comparatively less than in other dried vegetable and fruit. The niacin values of
dried apricot, cranberry, fig, peach, apple, seaweed and mashed potato were
2.5, 0.9, 0.5, 1.5, 0.91, 12.8 and 0.4 mg/100 g, while only 0.024-0.08 mg/ 100 g
niacin were found in dried banana pseudostem samples (NUTTAB, 2010 &
CNF, 2010). Compared with Musa balbisiana, Musa acuminata has less
127
6. Effect of drying on the carbohydrate digestibility of
banana pseudostem
have varied gastrointestinal and metabolic properties (Englyst et al., 2007). The
recent decades, the traditional belief that carbohydrates such as starch are
believed that the digestibility of carbohydrates determines the place and form in
of carbohydrates has benefits for some aspects of sports nutrition, this is not
metabolic syndrome (Englyst & Englyst, 2005). Slowly digested and absorbed
small intestine and poorly metabolized like resistant starch, some of the
oligosaccharides and dietary fibre will enter the colon and the fermentable
128
carbohydrates can reduce the risk of cardiovascular disease, diabetes, obesity,
This chapter will provide values of rapidly digestible, slowly digestible and non-
129
6.1 Sugars
The fructose, glucose, sucrose and total sugar contents of the pseudostems
acuminata pseudostem (dried at 50 °C) are presented in Table 6.1. All values
Table 6.1 Mean fructose, glucose, sucrose and total sugar content of Musa acuminata
Different species
balbisiana
acuminata
different (P<0.05).
130
significant decrease (P<0.05) in fructose from 8.0% to 3.1% was observed in
and total sugar content as well. These results indicate that blanching affects the
fructose, glucose and total sugar values in Musa balbisiana pseudostem, while
Saldivar et al. (2010) postulated that there was a significantly lower sugar
content in soybean seeds, which were blanched in water, than those without
water blanching. This could be attributed to some of the soluble sugars leaching
into the blanching water. In order to prove this assumption, the sugar contents
in the blanching water were detected by HPLC in the present study. A loss of
8.2 mg glucose/ 100 g fresh sample and 4.6 mg of fructose/ 100 g fresh sample
temperature and blanching in this study. BP50 had the lowest sucrose content
(3.9%) compared with the other three drying conditions. The sucrose content of
BP40, BP40B and BP50B were 4.4%, 4.9% and 4.5%, respectively. There was
water (Gamboa- Santos et al., 2012). Thermal destruction may be the cause of
of the cell wall, as well as disrupts subcellular organelles and their contents
131
become free to interact within the cell (Fellows, 2002). Another possibility is that
blanching stops the enzyme reactions in the banana pseudostem, which deters
Glucose, sucrose and fructose are metabolized differently. It has been stated
response than sucrose and glucose (Asp, 1995; Melanson et al., 2007). As a
especially in diabetes. Fig. 6.1 shows the ratio of single sugars in Musa
balbisiana pseudostem.
Figure 6.1 Effect of drying condition on the ratio of fructose, glucose and sucrose of Musa
balbisiana pseudostem
BP40B and BP50B have a similar ratio. This implies that drying
132
temperature has no influence on the change of single sugars ratio in Musa
balbisiana pseudostem, while blanching affects the ratio. The glucose ratio in all
standpoint. With the same drying conditions, different banana species provided
Figure 6.2 Effect of banana species on the ratio of fructose, glucose and sucrose
fructose, 9.4% glucose and 18.9% total sugar), Musa balbisiana (1.8% fructose,
133
4.0% glucose and 9.2% total sugar) has a higher value of fructose, glucose and
Musa acuminata. The sugar values of the two banana species significantly differ
(P<0.05) from each other. The glucose to total sugar ratio in these two species
is similar, but the fructose to total sugar ratio and sucrose to total sugar ratio
The sugar content of the banana pseudostem is similar to the range of most
vegetables. The fructose content of the banana pseudostem ranges from 0.1 g
/100 g to 0.7 g /100 g on a wet weight basis. This range of fructose is similar to
that of most vegetables, such as raw artichoke (0.3 g /100 g), fresh raw green
bean (0.1 g/100 g), fresh raw broccoli (0.2 g/100 g), raw celery (0.5 g/ 100 g),
raw peeled garlic (0.6 g/100 g) and raw red skin peeled pumpkin (0.8 g/100 g)
(NUTTAB, 2010), but lower than most fruits like cherry (5.4 g/ 100 g), banana
(4.9 g/ 100 g), pineapple (2.1 g/ 100 g) etc. (CNF, 2010). The glucose value of
artichoke (0.4 g / 100 g), fresh raw green bean (0.4 g/ 100 g), raw celery (0.7 g/
100 g), raw peeled garlic (0.4 g/ 100 g), raw cherry tomato (1 g/ 100 g) and raw
red skin peeled potato (0.9 g/ 100 g) (NUTTAB, 2010). The sucrose levels in the
banana pseudostem ranged from 0.2 g/ 100 g to 0.5 g/ 100 g, which is similar
value to artichoke (0.2 g / 100 g), fresh raw green bean (0.2 g/ 100 g), fresh raw
broccoli (0.1 g/ 100 g), raw peeled garlic (0.5 g/ 100 g), raw peeled pumpkin
(0.2 g/100 g), as well as muscadine grapes (0.6 g/ 100 g), raw sweet cherries
(0.2 g/ 100 g) and green kiwifruit (0.2 g/ 100 g) (NUTTAB, 2010 & USDA, 2007).
134
6.2 Starch
Starch content of the banana pseudostem in this study has been detected by
two methods, including HPLC method and Englyst method (1992). Results are
Table 6.2 Rapidly digestible starch, slowly digestible starch, resistant starch and total starch
(Englyst Method & HPLC Method) of Musa acuminata and Musa balbisiana
Musa balbisiana
Musa acuminata
The HPLC method determined the total starch content of the banana
pseudostem by digesting into glucose; while Englyst method not only detected
the total starch content but also categorized starch further into rapidly digestible
starch (RDS), slowly digestible starch (SDS) and resistant starch (RS),
135
which provides more information about the digestibility of starch. The starch
The total starch content of the blanched samples was 32.8% and 31.7% in
temperature, the sample with blanching had a notably higher total starch
The result above indicates that the drying temperature had no effect on total
Harvested, bananas have the ability to convert starch into sugar, such as
glucose and fructose (Dadzie &Orchard, 1997). The enzymes in the cell wall,
starch in the plant (Paliyath et al., 2009). Since the samples used in this study
were banana pseudostems, it is probable that the samples have some similarity
in properties to the banana fruit. Due to the heating effect of blanching, which
can destruct enzymes as well as damage the cytoplasm and other membranes
(Grandison, 2006), the reactions degrading starch into sugar will be stopped by
136
blanching. By contrast, samples without blanching have enzymatic reactions
that are likely to degrade starch further into glucose and fructose. As a result,
the starch contents of BP40 and BP50 are significantly lower than BP40B and
BP50B, but the fructose and glucose contents of BP40 and BP50 are
dramatically higher than BP40B and BP50B. This is one of the possibilities of
why blanched samples have higher starch values, but lower glucose and
fructose values. Further studies are required to prove that the enzymatic
The RDS, SDS, RS and TS contents of Musa balbisiana and Musa acuminata
by Englyst method are presented in Table 6.2. All values are on a dry weight
basis.
A highly significant increase (P<0.05) in RDS and SDS was noted in BP50B.
Compared with BP50, the RDS in BP50B rose over 100% from 7.7% to 16.4%.
The resistant starch in BP40B is significantly higher (P<0.05) than the other
samples detected by Englyst method with the same blanching conditions (with
each group of samples with the same temperature, blanched samples had a
dramatically higher total starch content than that without blanching. This trend in
total starch was the same as that observed in using the HPLC method. A similar
137
RDS, SDS, RS ratio was found in BP40 and BP50, however, the ratio observed
Figure 6.3 Effect of drying conditions on the ratio of rapidly digestible starch (RDS), slowly
digestible starch (SDS) and resistant starch (RS) of Musa balbisiana pseudostem
but also affects the ratio of RDS, SDS and RS in the banana pseudostem.
BP50B has the highest RDS portion, which is up to 50%, and lowest RS level,
which is only 22.9%. In contrast, BP40B has the lowest RDS ratio (30.2%) and
138
BP50B. According to the ratio of RS, SDS and RDS of the four samples, the
The effect of banana species on the fraction of RDS, SDS, RS and TS is shown
in Table 6.2. Despite SDS of Musa acuminata being lower than Musa
balbisiana, other fractions of starch and total starch content are higher than
differences (P<0.05).
139
It has been found in Figure 6.4 that the ratio of RDS of both banana species is
almost the same, but the ratio of RS of Musa acuminata is remarkably higher
than Musa balbisiana and the ratio of SDS is considerably lower than Musa
balbisiana. Therefore, the banana species has a great effect on the starch
content of pseudostem.
Figure 6.4 Effect of banana species on the ratio of rapidly digestible starch (RDS), slowly
140
Compared with other food sources, banana pseudostem has a higher level of
RS. Table 6.3 shows the RDS, SDS, RS and TS content of some cereals, fruits
and vegetables.
Table 6.3 RDS, SDS, RS and TS contents in some carbohydrate- containing foods
Wholemeal bread 56 4 1 61
Corn flakes 73 2 3 78
Potato biscuit 23 17 15 55
Banana flour 3 15 57 75
Spaghetti (cooled) 33 42 4 79
cold)
Maize RS product 55 9 34 98
min)
Bean flakes 27 16 6 49
As shown in the table, boiled potato (hot and cold), cold boiled millet and bean
of the whole-meal bread, corn flakes and cooled spaghetti were lower than the
pseudostem samples, while the RS value of potato biscuit, banana flour and
141
pseudostem is a good source of resistant starch, because though its RS content
was not the highest, the ratio of RS and SDS was relatively higher than other
Table 6.4 RDS, SDS, RS and TS contents in some carbohydrate- containing foods
The RS content of potato biscuit is nearly double that of the pseudostem, but
RS is notably less and RDS is more than the pseudostem. Meanwhile, the
fraction of RDS level of Maize RS product is more and the SDS is less than
pseudostem. Among the foods displayed in the tables, only banana flour had
142
significantly higher content and ratio of RS to RDS than banana pseudostem.
Partial least squares (PLS) were the regression method selected to relate total
starch content using Englyst method in comparison to the HPLC method. Linear
starch (Y = 0.97X -0.22) gave an intercept significantly different from 0.0 (P >
0.5), but the slope showed no significant difference from 1.0 (P < 0.5) and R2 of
Figure 6.5 Plot of HPLC method versus Englyst method for total starch of banana pseudostem
The results obtained for total starch by HPLC and Englyst methods were
similar, though these two methods were not perfectly related with each other.
Thus, other in vitro methods, such as the Nutriscan artificial gut can be used to
determine the RDS, SDS, RS and TS, to validate the two methods. Most of the
143
data obtained using the Englyst method was lesser than the HPLC method,
except for BP50B (Figure.6.6). The difference is more likely due to analytical
errors.
Figure 6.6 Total starch content of banana pseudostem detected by HPLC method and Englyst
method
Dietary fibre is composed of total dietary fibre (TDF), which includes both
soluble (SDF) and insoluble dietary fibre (IDF). Moreover, dietary fibre is
composed of different sugar fractions. The sugar contents within the soluble
144
and insoluble fibre contents. Three methods, namely the Megazyme method
145
6.3.1 Enzymatic-gravimetric method (Megazyme method)
The Table 6.5 illustrates the TDF, SDF and IDF of banana pseudostem with
different drying conditions and species using Megazyme total dietary fibre kit
Table 6.5 Mean SDF, IDF, TF of Musa balbisiana pseudostem and Musa acuminata
pseudostem
different (P<0.05).
All values are based on a dry weight basis and means of replicate.
146
6.3.1.1 TDF
The total dietary fibre contents of BP40, BP40B, BP50, BP50B and BPA were
19.2%, 18.5%, 21.5%, 24.0% and 15.89%. Drying at 50 °C with blanching had a
significantly (P < 0.05) high dietary fibre content. There was no significant
(P>0.05) difference between BP40 and BP50, as well as BP40 and BP40B, but
there was a significant difference (P <0.05) between BP40B and BP50. From
the result, both blanching and drying temperatures seemed to have effect on
significantly lower value of total dietary fibre than Musa balbisiana. Species had
The total dietary fibre content in this study was similar to that of Bhaskar et al.
(2011) (28.8%). All of the banana pseudostem samples contained much higher
levels of fibre than dried bulgur, unprocessed oat bran, dried apple and
2010). This result indicated that dried banana pseudostem has the potential to
6.3.1.2 IDF
The dominant portion of fibre in all banana pseudostem samples was insoluble
fibre. The content of insoluble fibre in BP40, BP40B, BP50, BP50B and BPA
was 15.6%, 15.2%, 17.8%, 20.3% and 11.2%, respectively. BP50B has a
notably high value of insoluble dietary fibre (P < 0.5), but no significant
difference was observed between BP40, BP40B and BP50. The IDF of Musa
147
dietary fibre in the banana pseudostem could promote intestinal regulation by
similar fibre containing foods (Jenkins et al., 2001, Cui & Roberts, 2009). Hence
6.3.1.2 SDF
Soluble dietary fibre in BP40, BP40B, BP50, BP50B were 3.5%, 3.3%, 3.6%
fibre between these four conditions. Though the total dietary fibre in banana
pseudostem reported by Bhaskar et al. (2011) and Aziz et al. (2011) were both
higher than in this study, the soluble dietary fibre content detected in this study
is higher (1.40% and 1.89%, for Bhaskar et al., 2011 and Aziz et al., 2011,
respectively). The dietary fibre analysis in this study implied that dried banana
oat bran. In order to know the properties of the banana pseudostem and to use
it for developing food products, further studies should be done to estimate the
(70.2%). As labeled, Benefibre contains 75% soluble dietary fibre, and this
148
6.3.2 NMR method
Solid state NMR and solution state NMR were used to explore the structure of
determined by NMR and hence only BP50 was used for this study.
The solid state NMR technology had been used to determine the structure and
characterization of plant cell wall polymers in apple, kiwifruit, lemon and maize
(Irwin et al, 1984; Newman, Ha & Melton, 1994; Newman & Redgwell, 2002).
Figure 6.7 a: 13C CP/MAS NMR spectrum of soluble dietary fibre of dried banana pseudostem
b: 13C CP/MAS NMR spectrum of insoluble dietary fibre of dried banana pseudostem
149
of acetyl groups in acetylated samples were also situated in this region. The
The peaks at 67-72 ppm came from the other carbons of pyranoid ring. The
which can be used for the estimation of degrees of methylation. The resonance
at 21 ppm represents methyl carbons of the acetyl ester OCOCH3, which can
be used for the estimation of acetylation values (Synytsya et al., 2003). The
chemical shift in the dried banana pseudostem soluble dietary fibre fraction was
compared with the chemical shift of the commercial pectins in the literature as
other compounds in the soluble dietary fibre could not be elucidated using this
150
method.
cellulose observed from the literature were the peaks at 105.5 ppm for C1,
peaks at 84.9 and 89.4 for C4, peaks at 62.5 and 65.4 for C6, and peaks at 73.0
and 75.0 for C2,3,5 (Ratnayake et al., 2011). Compared with Figure 6.9, similar
spectrum was observed in Figure 6.7b. As a result, the insoluble dietary fibre of
The weaker signal at 21 ppm in the figure was assigned to the acetyl group. By
this method, it could be found that cellulose was the major compound in the
Figure 6.9 CP/MAS 13C NMR spectrum of cellulose isolated from switchgrass from literature
1
H NMR spectroscopy is a valuable technique of characterization and
151
classification of dietary fibre structural features. This study used the Chenomx
NMR Suite library to fit the 1H NMR spectroscopy and to identify the sugar
fractions in the banana pseudostem dietary fibre. Figure 6.10 showed the1H
Figure 6.10 1H NMR spectroscopy of banana pseudostem total dietary fibre extraction
Black peaks represented spectroscopy detected by NMR and red peaks were
spectroscopy fitted by Chenomx NMR Suite. As shown in Table 6.6, the main
152
Table 6.6: Chemical shifts and portion of sugar fractions in the banana pseudostem dietary fibre
extraction
Although the 1H NMR can detect the sugar fractions in the dietary fibre and give
a high signal to noise ratio in a short experimental time, which reflects the
properties of the dietary fibre within several minutes, this method has its
drawbacks. It suffers from severe signal overlaps due to its short chemical shift
confirmed, due to the limitation of the library. For example, in Figure 6.10 the
technique, which cannot be used for accurate quantification of the neutral sugar
contents of dietary fibre. Hence for quantification of the fibre components, a gas
153
6.3.3 Enzymatic-chemical method (Englyst Method)
(1994). The GC analysis was by the same method with following modifications:
multi-point calibration; 3) Split injection was chosen for better separation. This
method identified and quantified the individual constituent sugars. This method
has also been described to be useful in studies where the relationship between
intakes of NSP and health as well as the relationship between NSP and its
physico-chemical properties (Englyst & Geoffrey, 1996; Kumar et al., 2012) can
the NSP result of banana pseudostem are presented in the following sections.
acceptable for its intended purpose (Green, 1996). In order to provide accurate
and reliable data from GC for NSP analysis, the linearity, limits of detection and
154
6.3.3.1.1 GC chromatography
The GC-FID separation was carried out using a Supelco SP-2380 wide-bore
and 275 °C for inlet and detector, which results in a good resolution and
within 9 min. A typical chromatogram is shown in Fig 6.11, and the elution times
and peak areas are listed in Table 6.7. The peaks for all neutral sugars were
sharp and symmetrical. It must be noted that good separation of the peaks are
related to 1) the ratio of the combination of the standard sugar mixture and
derivation; 3) dilution factor of split injection. Fig 6.12 shows the chromatogram
of standards with a wrong combination ratio (a) and with stale ammonia solution
(b) as examples.
155
a
156
a
Figure 6.12 Chromatogram of (a) standard with wrong combination ratio of standards to internal
157
Table 6.7 GC data of monosaccharides in standard mixture and banana pseudostem samples
Glu = D-(+)-Glucose
Allose was chosen as internal standard (IS) since it was eluted within the time
that all sugars were eluted and it did not co-elute with any of the tested sugars.
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6.3.3.1.2 Linearity
The calibration curves were obtained by plotting the peak area ratio between
the derivatives of neutral sugar standards and that of allose (IS) against their
levels of sugar mixture with 1 mg/ml internal standard as described in Table 6.8.
The reasons for choosing the calibration concentration at these levels were: 1)
the ratio of neutral sugars in each level was similar to the ratio in real food
provided excellent separation of the peaks; 3) the sugar values of test samples
the linear range, regression equation and correlation coefficients are shown in
159
Table 6.9. The calibration plots showed satisfactory linearity for all sugars as
the coefficient of determination (R2) were all higher than 0.99, which ranged
Table 6.9 Sensitivity and linearity characteristics of neutral sugars determination in the presence
measuring the magnitude of the analytical background response, plus 3.3 and
10 fold of the mean background signal ratio, respectively. The values obtained
are listed in Table 6.9. The LOD varied between 6.0 mg/L (Fuc) and 76.7 mg/L
(Man).
160
mg/L and 230.0 mg/L for optimal quantification.
standard deviation (RSD). The data are presented in Table 6.10. The precision
of banana pseudostem sample. According to the RSD values obtained for the
same sugar mixture sample, which was injected seven times repeatedly on the
RSD values from 0.12 to 4.35%, indicating that the consistency of the
monosaccharide level injected into the GC system was within 5%. The results
show good repeatability for the optimized method. The inter-day precision was
of the random errors between different extractions and days. The RSD values
between days range between 0.17% and 5.67%, which is less than 6%.
As shown in Table 6.10, the recovery rate of the sugars in the intra-day
experiment range from 97.39% to 102.73%, while the recovery rate in the inter-
day experiment range between 96.74% and 104.71%. The recoveries of both
inter-day and intra-day experiment are 100± 5%. This implies that the method is
deemed to be accurate.
161
Table 6.10 Inter-day (over a period of 5 consecutive days) and intra-day (n=7) precision and
Fuc 120 119.52 ± 1.72 1.44 99.6 119.84 ± 1.08 0.90 99.87
Ara 1188 1185.32 ± 4.14 0.35 99.77 1181.9 ± 1.99 0.17 99.53
Xyl 1113 1129.22 ± 3.16 0.28 101.46 1132.60 ±3.51 0.31 101.81
Man 920 916.24 ± 25.28 2.76 99.59 913.46 ± 32.12 3.50 99.29
Gal 705 707.00 ± 8.70 1.23 100.28 701.95 ± 5.63 0.80 99.57
Glu 2350 2351.26 ± 3.53 0.15 100.05 2350.75 ± 4.03 0.17 100.03
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6.3.3.2 NSP of banana pseudostem
were found to be in the range of 40.67-47.21 g/100g of dry sample (Table 6.11).
conditions (p>0.05). All samples analyzed contained less soluble NSP than
different conditions ranged from 10.71 to 17.47 g/100 g on a dry weight basis
and insoluble NSP content varied from 26.37 to 36.50 g/100 g on a dry weight
samples dried at 40 °C. Although the value of total NSP detected by the GC
method was higher than that detected by the AOAC method, the trend showed
that drying at higher temperature had slightly higher total NSP using both
methods. Samples dried without blanching had significantly high soluble content
drying and blanching did not have an effect on the total value of the NSP.
However, through thermal treatment, the soluble and insoluble fractions may
vary because they can be converted into more insoluble compounds. High
soluble NSP. The insoluble/ soluble NSP ratio varied from 1.5 to 3.4 and the
Figuerola et al. (2005), fibre sources have an insoluble/ soluble dietary fibre
ratio close to 1:2 which is suitable for use as a food ingredient. Furthermore,
163
was important for health properties and also for technological characteristics. A
BP50B was 1.5, 2.5, 2.0 and 3.4 respectively. Although the ratio of all the
samples was close to 2.0, drying at 50°C without blanching is the optimal
colour and used less drying time with less energy consumption. As a result,
The total NSP of Musa balbisiana and Musa acuminata dried at similar
NSP and lower insoluble of NSP than Musa balbisiana (p<0.05). The soluble
NSP content of Musa balbisiana and Musa acuminata were 33% and 41%
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Table 6.11 Distribution of neutral sugars (g/ 100 g of dry sample) in the soluble, insoluble and total fractions of NSP from banana pseudostem
BP40B Soluble NSP 0.12b±0.01 0.10c±0.01 0.74c±0.09 1.01b±0.10 3.21c,d±0.28 0.99c,d±0.19 4.35c±0.68 10.53b 0.98a±0.02 11.51b
Insoluble NSP ND ND 1.98b±0.07 4.06c,d±0.05 4.11b±0.45 1.07b±0.00 17.64c±0.69 28.85c,d 0.31a±0.01 29.13c,d
Total NSP 0.12b±0.01 0.10c±0.01 2.71b±0.16 5.07b,c±0.15 7.33c±0.73 2.06c±0.19 21.99c±1.66 39.38b 1.29a±0.03 40.67b
BP50 Soluble NSP 0.14b±0.00 0.11b,c±0.00 1.07b,c±0.01 1.42b±0.07 3.26a,b±0.04 1.27a,b±0.02 7.25a,b±0.77 14.52a 0.92a±0.02 15.44a
Insoluble NSP ND ND 1.71b±0.02 4.83b,c±0.13 4.52b±0.14 0.95b±0.02 19.18b±0.31 31.18b,c 0.27a±0.02 31.46b,c
Total NSP 0.14b±0.00 0.11b,c±0.00 2.78b±0.02 6.25b±0.05 7.78c±0.09 2.22c±0.01 26.43a±1.03 45.71b 1.19a±0.04 46.90b
BP50B Soluble NSP 0.14b±0.01 0.12b±0.01 0.99b,c±0.16 1.16b±0.12 3.77b,c±0.29 1.17b,c±0.16 2.48c,d±0.05 9.82b 0.89b±0.08 10.71b
Insoluble NSP ND ND 1.79b±0.00 5.14b±0.14 4.37b±0.36 1.01b±0.00 23.98a±1.84 36.30b 0.20b±0.03 36.50b
Total NSP 0.14b±0.01 0.12b±0.01 2.78b±0.16 6.30b±0.26 8.14c±0.65 2.18c±0.16 26.46a±2.64 46.12b 1.09b±0.11 47.21b
Banana pseudostem with different species
Musa Soluble NSP 0.14b±0.00 0.11b,c±0.00 1.07b,c±0.01 1.42b±0.07 3.26a,b±0.04 1.27a,b±0.02 7.25a,b±0.77 14.52b 0.92a±0.02 15.44b
balbisiana Insoluble NSP ND ND 1.71b±0.02 4.83b,c±0.13 4.52b±0.14 0.95b±0.02 19.18b±0.31 31.18b,c 0.27a±0.02 31.46b,c
Total NSP 0.14b±0.00 0.11b,c±0.00 2.78b±0.02 6.25b±0.05 7.78c±0.09 2.22c±0.01 26.43a±1.03 45.71b 1.19a±0.04 46.90b
Musa Soluble NSP 0.15b±0.01 0.13a±0.00 1.04b,c±0.06 1.15b±0.11 5.53a±0.45 1.46a±0.07 7.45b±1.50 16.90a 0.63b±0.07 17.53a
acuminata Insoluble NSP ND ND 1.69b±0.01 2.87e±0.02 4.69b±0.02 1.02b±0.00 14.84d±0.26 25.11d 0.32a±0.02 25.43d
Total NSP 0.15b±0.01 0.13a±0.00 2.72b±0.05 4.02c±0.08 10.22b±0.46 2.48b±0.07 22.28c±1.14 42.00b 0.95b±0.09 42.95b
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6.3.3.2.2 Sugar composition of soluble, insoluble and total NSP
The contents of total sugars, uronic acid, sugar composition of the soluble,
insoluble and total NSP isolated from banana pseudostem are depicted in
Table 6.11. The total sugar content in the total, soluble and insoluble NSP of
(500C) had significantly higher (p<0.05) insoluble NSP content than when
dried at low temperature and samples without blanching had significantly high
different temperatures. The total uronic acid content varied from 1.09 to 1.29
significantly (p<0.05) lower uronic acid than banana pseudostem dried using
significantly high insoluble NSP and uronic acid and low soluble NSP contents
were detected from Musa balbisiana. Uronic acid contents in the soluble
fraction were dramatically higher than that in the insoluble fraction in all
samples. This may be because uronic acid is one of the most important
166
Glucose, Mannose and xylose were the dominant neutral sugars in the
soluble, insoluble and total NSP fractions of the banana pseudostem samples
6.11. Glucose occupied 53-60% of the total neutral sugar content in banana
pseudostem, which had similar value with that estimated by NMR (58.7%).
which agrees with reports from other authors (Aziz et al., 2011; Cordeiro et al.,
2004 & Aisah et al., 2013). Results from this study were similar to those from
The cellulose microfibrils are remarkably distinctive features of the cell walls of
all plants since it may be associated with water and matrix polysaccharides
glucomannans (Kumar et al., 2012). β-glucan not only has the viscosity
enhancement property, but also has an elastic gel network formation property.
Wood & Webster (1986) stated that β-glucan can be utilized as thickening
For example, it has cholesterol lowering effects (Kahlon et al., 1993) and
167
mannose and glucose may relate to the hemicellulose family such as
are water soluble and have the ability to 1) lower glucose absorption from the
intestine; 2) lower the total plasma cholesterol and triglycerides and control
weight due to the satiation feeling produced by the filling of the intestine with
mannan gel (Kumar et al., 2012). Xylose occupied 12-14% of the total neutral
NMR (9.17%). High amounts of glucose and xylose could be due to neutral
with the cellulose microfibrils (Knudsen, 2014). It has been reported that
6.3.3.3 Comparison of NSP of banana pseudostem with commercial dietary fibre supplement
Two commercial dietary fibre supplements were tested in this study. They
were Metamucil natural granular powder (Procter & Gamble, USA) and
Benefiber (Novartis, Australia) powder, which are the most common fibre
168
completely in water, nearly all of the NSP in Benefiber were removed with
sugar and starch during the NSP precipitation procedure. As a result, the NSP
Metamucil had a significantly higher insoluble and total NSP than banana
(Table 6.11). However, the insoluble/ soluble dietary fibre ratio of the
Metamucil was 4.7, which was far more than the ratio of banana pseudostem
(1.5-3.5). This difference in insoluble/ soluble dietary fibre ratio may be due to
the different types of the NSP source. The main component of Metamucil is
that fibre extracted from fruits and vegetables contain a considerably higher
proportion of soluble dietary fibre, while cereal fibre has more insoluble fibre.
considered as a good source for dietary fibre. The insoluble/ soluble dietary
Metamucil. If the dietary fibre of the banana pseudostem was extracted and
food applications, such as in weight control diets due to its ability to retain
169
the consumption of insoluble dietary fibre.
pseudostem. The main neutral sugars in Metamucil were xylose (38% of the
total neutral sugar), followed by arabinose (23%) and mannose (21%), while
60%), mannose (17-19%) and xylose (12-14%). Rhamnose and fucose were
arabinogalactans. This agrees with Englyst and Englyst (2005) that xylose is
viscosity, good water holding capacity and good emulsion ability (Kumar et al.,
2012; Michniewicz et al., 1992). Additionally, Biliaderis et al. (1995) stated that
170
A survey was done on the popular dietary fibre supplements sold in the
supplements sold in the market were made from cereals and only one
(Normafibe Natural fibre daily supplement) among the six was a fruit dietary
fibre.
dextrin
daily supplement
soluble dietary fibre and better insoluble/ soluble ratio than cereals. The fruit
and vegetable based dietary fibre also contains different proportions of neutral
sugars in NSP from cereal based NSP, which provide different functionality.
Developing fruit and vegetable based dietary fibre supplements may meet the
demand of consumers who may require it to improve satiety rather than only
171
being used for fecal bulking. Banana pseudostem is a by-product of banana
fruit and its price is low. Based on its ideal NSP ratio, banana pseudostem has
and purity of the NSP fraction in banana pseudostem should be the next step
the characteristics of the dietary fibre. Three methods were used in this study
linkages of the pseudostem fibre, NMR was used. Although NMR is a quick
accurately. Furthermore, due to the limitation of the library, some of the sugars
quantify uronic acid and neutral sugars within the non-starch polysaccharides
172
component. The difference in the sugar composition and glycosidic linkages
The results from GC method were similar to that from the NMR method.
However, results from the AOAC method using the total dietary fibre assay kit
than that obtained from the GC analysis. One possibility is that the GC and
calibration method used in this study is different from that used in Englyst
method (Englyst et al., 1994). Nine level of calibration were selected according
to the NSP range of the banana pseudostem and commercial dietary fibre
supplement used in this study, while Englyst method used one point
calibration curve are more accurate than one point calibration. The operational
the real samples; this kind of calibration method can reflect the results much
closer to the real samples. From the study, it has been found that the results
from one-point calibration in the calibration range used in this study. Moreover,
the aim of the two methods is different, which causes the difference in the
method (985.29 & 991.43; AOAC, 2006) is to measure the sum of indigestible
173
method did not (Englyst et al., 2007). The AOAC method does not measure all
Both methods have their own benefits and disadvantages. AOAC method
gives values of TDF, SDF and IDF with good reproducibility which provides
enough information for routine analysis. However, it does not account for
analysis based on the structure and functionality of the dietary fibre. Difficulties
in the filtration step after enzymatic hydrolysis was seen in samples with good
Metamucil. When 1 g of Metamucil was used for the analysis, the Metamucil
formed a gel, which covered the celite layer and could not be filtered. Lowering
the sample size to 0.1g sample still proved difficult to filter which caused the
this method is not as time consuming as AOAC method when a large amount
1.5 days and 15 min is required for each sample on GC analysis. However,
Englyst method has a drawback as well. For example, it cannot determine the
value of dietary fibre samples with special processing, which makes the
174
hydrolysis. Hence, the NSP content of these samples cannot be determined
175
7. Effect of drying on the physicochemical properties
of banana pseudostem
Crystallinity index (CI), water holding (WHC), swelling (SWC) and solubility
different species are presented in Table 7.1. These properties suggest some
food products. For example, high WHC implies that the powder can be used
reflect its health benefits. For instance, high viscosity and solubility is able to
increase fecal bulking and reduce intestinal transit time, which may improve
intestinal peristalsis (Fleury & Lahaye, 1991; Gupta & Premavalli, 2011). The
176
Table 7.1 Physicochemical properties of dried banana pseudostem powder
WHC SWC SC CI
acuminata
balbisiana
different (P<0.05).
All values are based on a dry weight basis and means of triplicate
determinations.
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7.1 Water holding capacity (WHC)
under specified conditions of the temperature, time soaked and duration and
property that has been widely studied in food, since it is associated with food
dry matter. It has been reported that WHC of oat bran, rice bran, soy flour and
wheat bran fibre were 2.10, 4.89, 4.79-6.74 and 5.03 g water/g of dry matter,
respectively (Abdul-Hamid & Luan, 2000; Chen et al., 1988; Heywood et al.,
2002). The WHC content of banana pseudostem detected in this study far
exceeds those values from the literature. This implied that banana
pseudostem powder was able to bind or entrap more water than oat bran, rice
difference had not been observed between BP40 and BP50. Both drying
arabinoxylans, have good water holding capacity, which may hold up to 100 g
178
arabinoxylans are not bound to the cell walls, they can absorb about ten times
their weight of water (Choct, 1997). According to Chau and Huang (2003), the
proportion of cellulose in the NSP affected the WHC. Those foods containing
more cellulose would have lower WHC than those foods containing more of
physicochemical properties.
granules /g of dry matter). This indicated that banana species influenced the
SWC of the pseudostem. This finding agrees with the study by Mei et al.
than unblanched samples. Akter et al. (2010) and Aziz et al. (2011) stated that
blanching led to the increase of SWC, however this was opposite to the finding
in this study. One of the reasons is that other factors may also affect the SWC.
For example if the ratio of amylopectin and amylose is low in blanched banana
pseudostem powder, it may lead to the low SWC (Bello-Perez et al., 1999).
Thus, further studies need to be done to detect the amylose and amylopectin
difference (p>0.05) among BP40B, BP50 and BP50B, but BP40 has a
179
significantly higher value (p<0.05) of SWC than BP40B and BP50B. The range
matter. This value was similar to pea hulls and grape (5.2 g and 6.69of swollen
granules/ g of dry matter) and lower than carrot insoluble fibre, coconut fibre
and sugar beet fibre (18, 17 and 11 ml/g; Elleuch et al., 2011).
The solubility of BP40 was significantly higher (p<0.05) than other samples
relating to the SWC. The solubility was not affected by banana pseudostem
species.
Metamucil had significantly higher (p<0.05) SWC but lower solubility capacity
than banana pseudostem. This may be because the Metamucil has the
al., 2008). For example, the crystallinity of the banana pseudostem powder
may influence the flowability, bulk density, ease of handling, dust forming,
180
functions were assumed for each peak and a broad peak at around 21.5.
The patterns of XRD from Musa balbisiana dried at different conditions were
sharp peaks in the figures represent crystalline area, while the humps
were in the range of 13 - 25° and 32.5 – 40° with semi-crystalline peaks.
Despite these similar peaks, Musa acuminata had sharp crystalline peaks at
CI, while drying at 40 °C with blanching showed lowest CI. Blanching has no
effect on the CI. However, with the increase in drying temperature, the CI was
starch was destroyed gradually and more amorphous networks were formed.
species.
As shown in Figure 7.1 (c), Metamucil has more crystalline and less
pattern of the Metamucil was different from that of banana pseudostem. The
181
crystalline peaks of Metamucil were in the range of 7.5 – 27.5°. These
182
Crystalline
a
Amorphous
Crystalline
b
Amorphous
Figure 7.1 XRD results obtained from Musa acuminata (a), Musa balbisiana (b) banana
183
7.4 Pasting properties (RVA)
The pasting properties study using the Rapid Visco Analyzer (RVA) showed
viscosity (PV); through viscosity (TV); breakdown (BD); final viscosity (FV) and
set back (SB) between the banana pseudostem samples in different drying
conditions and species (Table 7.2). The pasting profiles of the banana
Figure7.2.
184
Table 7.2 Pasting properties of banana pseudostem (BP40, BP40B, BP50, BP50B, BPA) measured by Rapid Visco Analyser
PV (RVU) TV (RVU) BD (RVU) FV (RVU) SB (RVU) PEAK TIME (min) PASTING TEMP (C)
Different drying condition pseudostem
BP40 219.5a± 8.9 115.6a ±3.6 103.6a ± 7.1 146.8a ± 4.1 31.6b ± 1.2 4.2d ± 0.0 73.9c ± 0.5
BP40B 169.8b,c±4.3 95.5b ± 8.4 74.3b,c ± 1.2 112.8b ± 7.3 17.3c ± 0.2 4.5b ± 0.1 80.0a ± 0.1
BP50 170.0b,c ± 4.8 89.6b ± 2.6 80.4b ± 5.5 108.9b ± 3.7 19.3c ± 1.5 4.3c ± 0.0 74.4c ± .02
BP50B 188.9b ± 3.9 116.4a ± 5.3 72.5b,c ± 1.2 153.5a ± 4.6 37.0a ± 0.9 4.3c ± 0.0 77.1b ± 0.5
Different species pseudostem
Musa balbisiana
170.0b,c ± 4.8 89.6b ± 2.6 80.4b ± 5.5 108.9b ± 3.7 19.3c ± 1.5 4.3c ± 0.0 74.4c ± .02
Musa acuminate
157.3c ±3.5 100.3b ± 5.8 57.0c ± 0.2 117.8b ± 7.4 17.5c ± 1.8 4.7a ± 0.0 79.8a ± 0.2
Where: PV - Peak viscosity; TV – Through viscosity; FV - Final viscosity; BD – Breakdown; SB – Set back
All values are means ± SD triplicate measurements
Values in the same column with different superscripts are significantly different from each other (p< 0.05)
185
According to the figure, all of the differently treated banana pseudostem
samples dried at different conditions, BP40 had the highest PV, BD and FV,
while BP50B had the highest TV and SB. No significant difference (P > 0.05)
was observed in the value of PV, TV, BD, FV and SB of BP40B and BP50.
BP40B required longest time (4.5 min) to reach maximum viscosity. This might
be due to the lower rate of absorption and swelling of starch granules, which is
similar to the previous results as shown in Table 7.1. BP40B has the lowest
BP40 had significant higher (p<0.05) PV and required the shortest period of
time to reach the peak viscosity than banana pseudostem dried at other
conditions. This result implies that BP40 has the highest swelling power, which
agrees with the previous result (Table 7.1). The peak viscosity prior to
these swollen granules. Kim et al. (1999) reported that starch granules with high
swelling capacity occupied more volume in the water suspension, thus the
space between the granules became narrower and frictions among granules
were likely to occur. BP40 was assumed to have less structure rigidity during
BP40, which observed strong swelling capacity, had the highest PV and is easy
to reach maximum viscosity. This result suggests that BP40 would behave
differently during cooking and processing compared with samples dried in the
other three conditions. The soluble to insoluble NSP ratio is also significantly
186
a higher swelling capacity (Table 6.11; page 165). No significant difference (p >
0.05) was exhibited between PV of BP40B, BP50 and BP50B, and between
During the holding period of the viscosity test, the material slurries are
subjected to high temperature (95 C) and mechanical shear stress, which
further disrupt starch granules resulting in amylose leaching out and alignment
with high peak viscosities, which in turn is related to the swelling capacity of the
granules during heating (Ragaee et al., 2006). As shown in table 7.2, BP40 had
> 0.05) of BV. This result indicated that BP40 has poor paste stability when it is
Compared with Musa balbisiana, Musa acuminata had significantly lower (P <
0.05) BV. Musa acuminata pseudostem is more resistant against heat and
shear force than Musa balbisiana. This indicates that Musa acuminata may
structure and thereby viscosity will increase to a final viscosity (Ho et al., 2012).
The low SB values implies low rate of starch retrogradation and syneresis.
187
BP40B and BP50 showed lowest SB (17.3 RVU and 19.3 RVU, respectively)
followed by BP40 (31.6 RVU) and BP50B (37.0 RVU). This result indicated that
is an important indicator of the strength of the gel formed upon cooling and
BP40B and BP50 are significantly lower (p < 0.05) than BP40 and BP50B. No
188
8. Conclusions, recommendations and contributions
8.1 Conclusions
Two banana pseudostem species (Musa balbisiana and Musa acuminata) were
used in this study together with 4 drying conditions in order to figure out the
content, higher ash content, and no significant difference (p>0.05) in protein, fat
drying. This condition has been selected to drying both Musa balbisiana and
Musa acuminata for comparing the effect of species on the nutrients retention,
higher (p<0.05) moisture, protein and ash content than Musa balbisiana. There
189
percentage of total dietary fibre and resistant starch in samples dried at this
condition were the highest. Musa acuminata had lower digestibility than Musa
balbisiana. It has also been found that higher temperature (50 C) provided
higher level of total dietary fibre content. Moreover, blanching affected the sugar
and starch content in banana pseudostem. Higher starch and lower sugar
relationship was observed between total starch and sugars in the pseudostem
samples.
three methods were used and compared for determining the structure of dietary
fibre, namely AOAC, NMR and GC methods. According to the result, cellulose
was the main insoluble dietary fibre and pectin was the dominant soluble dietary
quantified. It has been identified that glucose, mannose and xylose were the
primary neutral sugars in the Metamucil were xylose, arabinose and mannose,
which were different from that in the banana pseudostem and resulted in
The WHC, SWC, SC and CI of Musa balbisiana were in the ranges of 9.0-11.0
g water/g of dry matter, 4.3-7.5 granules/g of dry matter, 25.5-34.2 % and 36.5-
190
than Musa acuminata. No significant difference (p>0.05) was observed in WHC,
significantly higher (p<0.05) PV and BD, which indicated that BP40 had the
highest peak viscosity and poor pasting stability. BP40B and BP50 had
Metamucil were significantly higher than banana pseudostem (p<0.05) and the
banana pseudostem.
8.2 Recommendations
191
development of pseudostem as a food ingredient.
(FID).
192
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10. Publications Derived from this Project
Ma, J., Srzednicki G., and Arcot J. 2015. Effect of Drying on the Dietary
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11. Appendix
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EFFECTS OF DRYING ON THE DIETARY FIBRE OF BANANA
PSEUDOSTEM (MUSA BALBISIANA & MUSA ACUMINATA)
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Jun Ma, George Srzednicki(*) and Jayashree Arcot
Food Science and Technology Group, School of Chemical Engineering, UNSW Australia, Sydney 2052
(*)
Email: g.srzednicki@unsw.edu.au
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ABSTRACT
This study used two methods to identify the dietary fibre in banana pseudostem. The first
method was using Megazyme (2012) to identify the total, soluble and insoluble fibre in
banana pseudostem of two species, Musa balbisiana and Musa acuminata. Pseudostem
samples were subjected to four drying treatments namely drying at 40 °C with or without
blanching and drying at 50 °C with or without blanching. Banana pseudostem drying at 50°C
with blanching resulted in significantly higher (p<0.05) insoluble and total dietary fibre than
banana pseudostem content than other treatments. No significant difference was observed in
soluble fibre of banana pseudostem between treatments or species. Musa balbisiana
pseudostem showed significantly higher total and insoluble dietary fibre value (p<0.05) than
Musa acuminata pseudostem. The second method was using enzyme and acid hydrolysis to
extract dietary fibre and break down the links to release single sugars and then used solution
state 600 MHz NMR 1H and solid state 13C CP/MAS NMR method to identify the
composition and network of dietary fibre in banana pseudostem. The results of the study of
fibre structure and value in the banana pseudostem characterise its physicochemical
properties, nutritional value and thus provide information on the utilisation of banana
pseudostem in the food industry that have potential to increase the nutritional and economical
value of banana pseudostem.
Keywords: Banana, Pseudostem, Dietary fibre, Drying, Blanching
INTRODUCTION
The stem of the banana plant, which is also called pseudostem, produces a single bunch of
banana before dying and replaced by a new pseudostem (Anhwange et al., 2011). Due to this
character, the crop generates a large amount of residue. It has been estimated that a few
tonnes of banana pseudostem per hectare are produced every year (Aziz et al., 2011). After
the harvest, the bare pseudostem is cut and usually left on the plantation or burned, which
could ultimately cause environmental issues and economic loss (Cordeiro et al., 2004). In
Australia, over 300,000 tonnes of bananas are produced annually and million tonnes of
pseudostems are wasted (Horticulture Australia, 2014; Hossain et al., 2011). Thus, the
utilisation of the banana waste - pseudostems and the processing methods to enhance their
economic value has gained more attention in recent years.
Nowadays, the banana pseudostems are being used as a raw material for paper, furniture and
forage (Buragohain et al., 2010; Umaz et al., 2005). In some regions such as India and
Malaysia, the fresh tender core of banana pseudostem is cooked and consumed, whereas the
consumption of banana pseudostem as food in Australia is rare. Some studies also state that
banana pseudostem is rich in minerals and nutrients especially dietary fibre (Cordeiro et al.,
2004; Ho et al., 2012; Aziz et al., 2011; Bhaskar et al., 2011). However, the studies on the
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