Accepted Manuscript

Effects of different drying conditions on the starch content,

thermal properties and some of the physicochemical parameters of
whole green banana flour

Amir Amini Khoozani, Alaa El-Din Ahmed Bekhit, John Birch

PII: S0141-8130(18)36477-8
DOI: https://doi.org/10.1016/j.ijbiomac.2019.03.010
Reference: BIOMAC 11840
To appear in: International Journal of Biological Macromolecules
Received date: 25 November 2018
Revised date: 15 February 2019
Accepted date: 2 March 2019

Please cite this article as: A.A. Khoozani, A.E.-D.A. Bekhit and J. Birch, Effects of
different drying conditions on the starch content, thermal properties and some of the
physicochemical parameters of whole green banana flour, International Journal of
Biological Macromolecules, https://doi.org/10.1016/j.ijbiomac.2019.03.010

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 ACCEPTED MANUSCRIPT

Effects of different drying conditions on the starch content, thermal

properties and some of the physicochemical parameters of whole green

banana flour

Amir Amini Khoozani*, Alaa El-Din Ahmed Bekhit, John Birch

Department of Food Science, University of Otago, Dunedin, New Zealand

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Postal address: University of Otago PO Box 56, Dunedin 9054

[email protected]

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Correspondence Authors:

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Amir Amini Khoozani

Email: [email protected]
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Prof. John Birch
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Email: [email protected]

Tel: +64220894289
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Abstract
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The physicochemical and thermal properties of whole green banana flour obtained from oven
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air-drying (ODF) at three temperatures (50, 80 and 110 °C) and freeze-drying (FDF) were

compared to wheat flour (WF). Lightness and yellowness were negatively affected by the
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temperature increment. The FDF samples exhibited higher a* and L* values and had the closest
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browning index to WF (P-value <0.01). Also, the ODF50 samples had the highest emulsion

activity, whereas FDF had the highest emulsion stability (P-value <0.05). The oil holding and

water holding capacities of the FDF samples were significantly higher than all other samples

(P-value <0.05). A higher resistant starch content was found in the FDF (46.72%) and ODF50

(44.58%) samples. The ODF50 samples had the highest amylose content and degree of

crystallinity (P-value < 0.05) as well as the lowest gelatinization temperature amongst the green

banana flour samples (4.69 °C). Oven air-drying increased the gelatinization temperature
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drastically for all GBF samples (P-value < 0.05). In conclusion, drying the whole green banana

at 50 °C resulted in the least negative effects on RS content compared to the other drying

temperatures investigated in this study and was the closest to the characteristics of the FDF

samples.

Keywords

Banana, drying, physicochemical properties, resistant starch, thermal properties

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1. Introduction

The increased demand for functional food products has led to an enhanced interest in whether

nutrients such as minerals, vitamins, bioactive compounds, fiber and prebiotics are present in

food formulations [1]. Amongst the prebiotics, RS has gained a lot of attention in recent years,

due to its acknowledged health benefits. Resistant starch is present in grains, legumes and seeds

(RS1), raw starchy plants (RS2), retrograded starchy foods (RS3), chemically modified starch

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(RS4) and in amylose-lipid complexes (RS5) that are considered to be physically inaccessible

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to digestion [2]. Banana, the world’s most favorite fruit, is one of the richest sources of RS2 at

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early stages of ripeness [3].

Globally, banana is reported to be the fourth most demanded food after rice, wheat and corn
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[4]. According to FAO, banana (Musa Cavendish, AAA group) is an essential part of New

Zealanders diet with a per capita consumption on average of 20 kg of banana per year, which
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is reported to be more than the worldwide average consumption [5]. However, banana is the

second most discarded food in New Zealand after bread, contributing to 3 % of the total food
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waste [6]. Banana is a perishable food commodity and is also sensitive to postharvest defects
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including bruising during transportation, browning due to inappropriate ripening environments

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and it is reported may even be rejected because of its “unacceptable” curved shape [7].
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Unripe banana is rich in RS which is recognized to have positive effects on colon health [8],

while ripe banana contains more digestible starch and protein [9]. Bananas consist of two parts,
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the banana pulp (BP) and the banana peel (BPe). The pulp is a rich source of essential

phytonutrients, phenolic compounds, B group vitamins, ascorbic acid and tocopherols, while

the BPe is a rich source of minerals, bioactive compounds and dietary fiber (DF) such as pectin,

cellulose, hemicelluloses and lignin [10].

Considering the nutritional value of both pulp and peel of green bananas, the production of

green banana flour (GBF), which can be obtained by drying, provides a way to preserve the
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nutritional benefits and increase the shelf-life of banana material [11]. The drying conditions

used, such as different temperatures and humidity levels, can have a profound influence on

both functional and technological characteristics of the obtained GBF. In addition, the way in

which green banana can be prepared for the drying process (whole, paste, slices) exerts

different properties on the final product [12-14].

Unripe banana flour is considered to be a suitable formatted product for use in food processing,

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in addition to its high RS2 content and low digestible starch [15]. Hence, the preparation of

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green banana flour requires a drying method that results in retention of nutritional properties,

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especially RS2.

The effects of room drying [16], including oven air-drying [17, 18], spouted bed with hot air
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flow [19], pulsed-fluidized bed agglomeration [20], ultrasound and pulsed-vacuum followed

by air drying [13], microwave heating [12], freeze-drying [12, 15] and spray drying [21] on
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some of the attributes of different green banana parts have been reported. However, the

collective information available comparing different oven air-drying temperatures with non-
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heat treatment processes on properties of unpeeled GBF, especially on RS content and

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hydration properties, have scarcely been reported in the literature. Also, the effect of freeze
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drying, which is considered to be one of the best dehydration methods for heat-sensitive
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materials, on the thermal properties and starch content of GBF has not yet been investigated.

Compared to other methods of drying, freeze-drying is considered to be the best dehydration

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method for heat-sensitive materials to enable achievement of the highest possible quality,

minimal comprising of color, structure, nutrients and flavor. As most of the leading banana

producers are located in developing countries, a simple and efficient drying method would be

preferable for production of GBF with the highest possible yield and quality. Consequently,

from the feasibility point of view in the food industry and the minimum effect on green banana

nutrients, oven air-drying and freeze-drying (followed by blast freezing) were chosen for
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analysis in this study, to investigate the effect of processing on the properties of the produced

flour.

This study aimed to evaluate the effect of freeze-drying and of three different oven air-drying

methods on whole GBF, with examination of the effects of these methods on the starch content,

functional behavior and some of the physicochemical properties.

2. Materials and Methods

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2.1. Sample preparation

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A total of 20 kg green banana (Cavendish of Musa (AAA) Nanica cultivar) was supplied by a

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local food supply company in Dunedin, New Zealand between February and March 2018. The

bananas were harvested approximately 100 days after anthesis and were not exposed to any
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maturation treatments. Samples of the bananas were processed on arrival and were used for the

measurement of pH, firmness, total soluble solids and titratable acidity. The production steps
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used to produce the dried green banana flour are shown in Fig. 1. Wheat flour was provided by

the same local food supply company in Dunedin New Zealand and was used as a reference
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sample.
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Figure 1. Flow chart of green banana flour production by two different drying methods
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2.2. Sample characterization

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The pH of processed banana samples were measured using a pH meter (HANNA Instruments,

Woonsocket, USA). Titratable acidity was determined according to the official method 942.15
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(2005) AOAC [22] using an analytical radiometer (Ph-STAT, Colorado, US). A hand-held

refractometer (ATAGO N1, California, USA) was used to determine the total soluble solids

with correction for acidity and temperature values according to Kar, Chandra, Prasad, Samuel

and Khurdiya [23]. Total solids were assessed by gravimetric determination after drying at 60

°C under a pressure of 60 mmHg for 16 hours.

2.3. Green banana flour production

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After rinsing the supplied green bananas with distilled water containing sodium hypochlorite

(10 g/l), the bananas were immersed in 0.5 % (w/v) citric acid solution for 15 min and then

drained, in order to reduce enzymatic browning. The green bananas were then cut into slices

with 2 mm thickness and then dipped in 0.5 % (w/v) citric acid solution for 15 min and then

drained well. The treated sliced bananas were dried using either an oven air-drying method or

freeze-drying method.

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2.3.1. Oven air-drying method

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The sliced bananas were distributed on screened trays and transferred into a pre-heated forced-

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oven air-dryer (LABEC, Laboratory Equipment Pty Ltd., Marrickville, NSW, Australia) at

either 50 °C, 80 °C, or 110 °C for either 2, 4 or 7 h, until no further change in weight was
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achieved.

2.3.2. Freeze-drying method

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The banana samples were placed on stainless steel trays in a blast freezer (IRINOX, Treviso,

Italy) at -30 °C for 4 h and then transferred to a freeze dryer (SP Scientific, New York, USA).
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The dried green banana slices that were subjected to either of the drying methods were then
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ground and the powder passed through a 70 mesh (212 µm) screen, then vacuum packed, and
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stored at 4 °C until used for analysis.

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2.4. Physicochemical properties of the banana flour

Some of the physicochemical properties, such as water activity (aw), moisture content, ash,
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protein, lipid and carbohydrate, were determined according to the AOAC 2000 standard

methods [22]. The aw was measured by an Aqualab device (Decagon Devices, Washington DC,

USA) at 24 °C. The moisture content of the samples was reduced using a vacuum oven dryer

(Thermoline, Australian Marketing Group, Marrickville, NSW, Australia) at 60 °C for 16 hours

(Method 925.40; AOAC 2000). The ash content was determined following ashing of samples

in a furnace (Method 923.03; AOAC 2000). The Kjeldahl method was used to determine the
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protein content (%N×6.25) of samples using Method 923.03; AOAC 2000. The lipid content

was determined using the Soxhlet extraction method (Method 920.39; AOAC 2000) [22]. The

carbohydrate content was calculated by the following equation:

Carbohydrate (g) = 100 − [moisture content (g) + ash (g) + protein (g) + lipid (g)].

2.4.2. Color measurements

The color of banana samples was assessed by using a Hunterlab Spectrocolorimeter (Hunter

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Lab Mini Scan Plus Colorimetric, USA). The color parameters were defined using the CIE

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L*a*b* system, where L∗ (L∗ = complete black and L∗ = 100 perfect white), a∗ (−a∗ = greenness

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and + a∗ = redness) and b∗ (−b∗ = blueness and +b∗ = yellowness) values were calculated [24].

Chroma values (C) that were near zero were interpreted to indicate subdued colors, whereas
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high chroma values were interpreted to specify a more vibrant color. Browning index and

chroma were measured by the following formula [25].

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Chroma = √a∗2 + b ∗2

100(x − 0.31)
Browning index = , where x is obtained using the following formula:
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0.17
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a∗ + 1.75L∗
x=
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5.645L∗ + a∗ − 3.012b ∗

2.4.3. Oil and water holding properties

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The oil holding capacity (OHC) and water holding capacity (WHC) were determined as
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previously described by Nasrin, Noomhorm and Anal [24]. The solubility index (SI) and

swelling power (SP) were determined according to the method of Nasrin, Noomhorm and Anal

[24]. One g aliquots of banana sample were dispersed in either 50 mL of distilled water to

determine the WHC, or 50 mL of canola oil to measure the OHC in previously weighed

centrifuge tubes (NALGENE®). The tubes were capped and placed in a water bath at various

temperatures (40 °C, 60 °C and 80 °C) for 30 min with intermittent stirring. The tubes were

then cooled to room temperature and then centrifuged at 1500 g for 20 min. The OHC was
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calculated as g of oil per g of dried flour. The supernatant generated from the centrifugation

step in the WHC measurement was dried at 105 °C for 5 h to determine the SI (g/100g). The

paste was weighed and then dried for 5 h at 100 °C. The WHC (g/g) and SP (g/g) were

determined according to the following formula [26]:

WHC = (W3 − W4 )/W4 , where W3 and W4 are the weights of wet residue and dry residue,

respectively.

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W2
SI = × 100, where W1 and W2 are the weights of the dry sample itself and the dry sample
W1

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in supernatant, respectively.

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W3
SP =
W4
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For determination of emulsion activity (EA), a mixture consisting of 10 mL of distilled water,

10 mL of soybean oil and one g of the sample was put in a calibrated centrifuge tube. After
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centrifugation at 1500 g for 5 min, the ratio of the height of emulsion layer to the total height

of the mixture in the tube was recorded as EA percentage [27]

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After keeping the emulsion at 80 °C for 30 min in a heated water-bath, the tubes were cooled
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down with cold tap water and then centrifuged at 1500 g for 15 min. The emulsion stability
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(ES) was calculated as the ratio of the height of the emulsified layer compared to the total
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height of the mixture in the tube [27]

2.5. Relative Crystallinity

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An X-Ray diffractometer (Bruker Model D8 Discover, Billerica, USA) was set up with

parameters of 30 mA and 40 kV, a diffraction angle (2θ) range of 5–40°, a 0.03° step size and

measuring time of 15 s, according to the method reported by Campuzano, Rosell and Cornejo

[3]. The relative crystallinity (RC) was calculated according to the following equation:
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Ac
RC(%) = ( ) × 100, where Aa is the amorphous area and Ac is the crystalline area. These
Ac+Aa

areas were quantified using the software Origin®2017 (OriginLab Corporation, Northampton,

USA).

2.6. Starch analysis

2.6.1. Resistant starch

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The RS content was determined according to the methodology of AOAC 2002.02, using a

Megazyme kit (Resistant starch assay kit; Megazyme International, Wicklow, Ireland)

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according to the manufacturer guidelines. The samples were incubated in a shaking water bath

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with two enzymes, pancreatic α-amylase and amyloglucosidase (AMG) for 16 h at 37 °C in

order to achieve complete hydrolysis of the non-resistant starch. After centrifugation, the pellet
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was washed with ethanol (50 % v/v) and the supernatant was decanted. The RS in the pellet

was dissolved in 2 M KOH by vigorously stirring in an ice-water bath for 20 min. The starch
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was quantitatively hydrolyzed to glucose with AMG and then the glucose determined using a

glucose oxidase/peroxidase reagent (GOPOD) and RS was calculated as mg of glucose × 0.9

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(as the conversion factor).

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2.6.2. Total Starch

The total starch content was assessed according to the AOAC method 996.11 using a kit (total
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starch assay kit; Megazyme International, Wicklow, Ireland). The principle of the method

involves the hydrolysis of starch using a thermostable α-amylase into soluble branched and
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unbranched maltodextrins, according to the method of Moongngarm, Tiboonbun, Sanpong,

Sriwong, Phiewtong, Prakitrum and Huychan [8]. Digestible starch (DS) was determined by

pooling and washing the supernatant, adjusting the volume to 100 mL, and measuring the D-

glucose content with GOPOD. RS was subtracted from TS to determine DS.

2.6.3. Amylose and Amylopectin

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The amylose content was quantified using a commercial kit (amylose/amylopectin kit;

Megazyme International, Wicklow, Ireland) based on the separation of amylose and

amylopectin and precipitation of amylopectin with the addition of concanavalin-A (Con A) and

removing it by centrifugation.

2.7. Thermal properties

Starch gelatinization temperature was examined by differential scanning calorimetry (Diamond

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DSC, Perkin Elmer, Connecticut, USA), using an instrument equipped with an intercooler unit

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(Perkin Elmer, Model 2P, USA.), with nitrogen as the purge gas. Based on the method of

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Tribess, Hernández-Uribe, Méndez-Montealvo, Menezes, Bello-Perez and Tadini [28],

approximately 2 mg (db) of the sample was accurately weighed in an aluminum pan and 7 µL
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of deionized water was added. After tightly sealing the pans and maintaining them at room

temperature for an hour, the samples were exposed to a temperature gradient from 24 °C to 120
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°C at a rate of 10 °C min-1. The gelatinization/ peak temperature (Tp), onset temperature (T0)

and transition enthalpy (ΔH) and gelatinization range (R) were obtained directly from the data
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curves using the Pyris® manager software.

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2.8. Statistical analysis

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Statistical analysis was performed by Minitab® program version 16. After confirming that the
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data were normally distributed, the ANOVA test was applied to assess the significance of the

differences among the treatments for each variable. The correlation between the variables was
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determined using Pearson’s correlation. The differences among the means were resolved using

Tukey’s test. The significance level was set at P-value < 0.05, with a confidence interval of 95

%. All the tests were performed in triplicates.

3. Results and discussion

3.1. Physicochemical properties

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The physicochemical characteristics and firmness of green bananas are presented in Table 1

compared to data reported by Tribess, Hernández-Uribe, Méndez-Montealvo, Menezes, Bello-

Perez and Tadini [28] and Abbas, Saifullah and Azhar [26]. The results in Table1 suggests that

the banana samples used for GBF production was at the first stage of ripeness.

Table 1. Physicochemical properties and firmness of green banana compared to

previous findings

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3.1.1. Proximate composition

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The composition of GBF flour samples compared to the reference control (WF) is shown in

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Table 2. Compared to WF, all GBF samples regardless of drying method showed significantly

lower moisture content (MC), aw, protein content and lipid content values, and significantly
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higher ash and carbohydrate contents (P-value < 0.05). This behavior was also reported in a

report on GBPF [29]. The lower water content in all GBF samples indicated better stability and
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longer shelf life due to the lower potential for microbial growth (aw < 0.4) and better stability

against physical and chemical reactions. Amongst the GBF samples, as the drying temperature
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increased above 50 °C, a substantial change in water content was observed (P-value < 0.05),
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which may be related to the higher rate of drying compared to others [30]. The FDF samples
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had the same carbohydrates, lipid, protein and ash contents found in oven-dried samples.
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However, the FDF samples had higher moisture content than the ODF80 and ODF110 (P-value

< 0.05) and was not different from ODF50 (Table 2).
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As green bananas have been considered as valuable fruits due to their abundance in minerals,

taking advantage of the minerals in the BPe also resulted in higher amount of ash compared to

peeled green banana flour [11]. The results indicated that the carbohydrates content was over

80 % of the GBF (g/100 g db), which was similarly found by Bezerra, Amante, de Oliveira,

Rodrigues and da Silva [29] who investigated on GBPF. The inclusion of the peel in addition
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to the pulp could lead to a greater amount of RS2 and RS3. Overall, the effect of drying methods

on the composition of GBF was not statistically significant (P-value > 0.05).

Table 2. The composition of green banana flour and wheat flour samples (db g/100 g)

3.1.2. Color

The differences in color parameters of GBF samples and WF are shown in Table 3. The results

showed that FDF had a light green bright flour; with the highest values for lightness,

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yellowness and chroma amongst the GBF samples. Increasing the temperature resulted in a

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darker color amongst oven dried samples (p < 0.05). The same trend was observed by previous

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researchers in GBPF [17, 31, 32]. The degradation of chlorophyll, which is sensitive to oxygen,

heat and pH, produces pheophytin and pheophorbide which accounts for the green–brown color
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[32]. Moreover, the high amount of starch and protein along with the high temperatures may

have resulted in the partial Maillard reaction [33]. Similarly, Yangilar [17] stated that FDF had
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a greener color than spray dried banana pulp flour. The degree of color saturation (chroma)

was notably higher in FDF compared to oven dried GBF and WF (P-value < 0.05) in the current
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study, which also corresponds with Aziah and Komathi [34] findings on unpeeled pumpkin
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four. Compared to the spouted bed drying method, GBF obtained from freeze-drying and all
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oven air-drying methods were darker in color [19]. Also, it was reported that GBF made from
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BPe had shown a darker color when produced by microwave drying and freeze-drying,

respectively [12, 15]. According to Jeet, Immanuel and Prakash [35], the bright color of flour
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could be more acceptable for the consumer. Yet, the incorporation of flour as an ingredient into

food products could eventually affect the color of the flour. In general, it seems that more starch

would be preserved in FDF and ODF50 compared to other samples.

Table 1. Color analyses of green banana flour and wheat flour

3.2. Hydration properties
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As shown in Table 4, as the temperature increased, the mean value of WHC increased

significantly in all GBF and control samples (P-value < 0.05). At 40 °C, all GBF samples

absorbed more water than WF, and the FDF samples showed the highest significant value (P-

value < 0.05). Similar trends among the samples were observed at higher temperatures with the

ODF110 values had lower values than WF at 60 °C and 80 °C. While WHC could be related

to the physical status of amylose [36], the amount of insoluble dietary fibers, protein and

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particle porosity of the flour could also play an important role in water absorption [17, 24].

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Table 4. Hydration properties of Green banana flour and wheat flour

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Increasing the temperature results in protein denaturation and starch gelatinization. This may

lead to greater water absorption due to the availability of hydroxyl (OH) groups [37].
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According to Alkarkhi, Ramli, Yong and Easa [38], the release of amylose into water during

heating, occurs more in pulp flour than peel flour. Compared to other findings, the current
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results showed higher WHC for FDF than those reported by previous findings [9, 17]. High

WHC indicates the capability of FDF to be used as a thickener in semi-liquid and liquid foods
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[26]. Also, Grigelmo-Miguel, Gorinstein and Martín-Belloso [39] stated the syneresis in food
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can also be controlled by adding flour with high WHC due to the increment in expansibility of
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starch matrix.
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The OHC of flour relates to the hydrophobic character of starch: the higher the OHC, the higher

the EA [27]. In other words, stabilization of food systems with high fat content, such as in
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bakery products, can be easily achieved by emulsifiers [40]. By promoting the formation of

starch-protein complex and strengthening the gluten-gluten interaction, emulsifiers can

improve the aeration in bakery products [34]. The results of this study specified a substantial

increase in OHC for FDF sample at all temperatures compared to the control and other GBF

(P-value < 0.05). Comparing results at 80 °C showed a significant increase in OHC of ODF50,

which means it can be considered as an emulsifier compared to the control and other GBF
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samples. However, ODF50 showed significantly lower values for OHC compared to FDF. The

current results for FDF were higher than those reported for green banana pulp flour (GBPF)

[17, 27], mango peel flour [41] and citrus peel flour [42]. The OHC found in FDF is almost

equals to the OHC content of Balady flat bread which contained GBPeF [16] and unripe

plantain peel flour [37].

It has been previously stated that the amylose-lipid complex formation, which can be formed

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during heating at above gelatinization temperatures, produces RS5 which is resistant to

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hydrolysis formulation [43]. As a result, FDF could increase the RS content, and also be used

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as an alternative emulsifying agent in food. The results of the emulsifying properties were

correlated with OHC properties (r > 0.92, P-value < 0.05). The ES of different samples ranged
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between 30.54 % and 36.47 %, where ODF110 had the lowest stability and ODF50 showed the

highest stability (P-value < 0.05) as shown in Table 5. However, the ES values varied from
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9.69 % to 28.99 % for WF and FDF, respectively. Consequently, FDF appears to act as an

emulsifier as it has the highest stability amongst whole green banana flours as well as WF.
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Table 5. Emulsifying properties of green banana flour and wheat flour

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The SI is related to the soluble solid contents in dried flour, while the SP is an index that relates
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to the strength of the starch granule to stay intact during high cooking temperatures [29]. The
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SI and SP specify the range of interaction within the amorphous (amylopectin) and crystalline

(amylose) areas of starch molecule together with the length of branches, degree of branching,
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and formation of the molecules [24]. As mentioned earlier, an increase in SI and SP brings

about gelatinization, which is the basis for making pre-cooked starchy foods, especially in

bakeries [44].

The obtained results showed that all GBF samples had higher SI compared to WF, and this

increase was significant for ODF80 and ODF110 (P-value < 0.05). However, the increment in

SI was not as high as GBPF [3, 29, 45], but greater than in green banana peel flour (GBPeF)
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[24, 46]. The same pattern was observed for SP as well; considering FDF had a more stable

starch structure than other GBFs as temperature increases. As Campuzano, Rosell and Cornejo

[3] reported, the ripening stage is positively correlated with SI. By increasing the temperature

over the gelatinization temperature (more than 65 °C), starch degradation and sugar release

lead to an increase in both SI and SP.

The distribution of amylopectin and amylose in starch granule was suggested as a factor that

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influenced the SI value by Seguchi, Tabara, Iseki, Takeuchi and Nakamura [47]. As amylose

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plays a key role in maintaining the starch structure, the higher amylose content could eventually

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create a more compact starch granule. At temperatures rose above 65 °C, both the SI and SP

were dramatically increased (P-value < 0.05). As the hydrogen bonds break, the water
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molecules bind to the exposed OH groups and the granule expands and exudes amylose,

meaning it would be more difficult for starch to overflow outside of the granules [45].
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According to Ratnayake, Hoover and Warkentin [46], green banana starches compared to those

of corn, chayote and cassava, swell more slowly, which may signify that a strong micellar
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arrangement needs to be broken. In consequence, incorporation of the peel into the pulp flour
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has improved the SI and SP of the whole GBF.

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3.3. Starch analysis

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Starch is classified into two groups based on its digestibility: digestible starch (comprises of

slow and rapidly digestible starches) and resistant starch [48]. In the whole green banana, 70-
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80 % of the dry weight contributes to starch, considering that the peel contains up to 12 % of

the total amount of starch [3]. It has been previously described that the RS type in green banana

is type two, which is resistant to hydrolysis [49]. However, during the production of GBF the

amount of RS2 decreases due to gelatinization, especially in higher temperature [23]. That fact

signifies the importance of choosing the right processing condition together with proper pre-

treatments.
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The present results corroborate the previous findings about the destructive effect of drying at

higher temperatures (Fig. 2). Interestingly, ODF50 displayed similar values to FDF, which had

the highest amount of RS. Although the difference between FDF and ODF50 samples was

significant (P-value < 0.05), the RS content was well preserved during the ODF50 heat-treated

flour compared to ODF80 and ODF110.

More than half of the total starch in FDF and ODF50 was RS. While the digestible starch

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represented a range between 17 % and 36 % dry weight base (db) in GBF samples, the WF

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contained the highest amount of available starch. Also, the results indicated that drying

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temperatures over 80 °C did not have a significant effect on RS content reduction (P-value >

0.05).
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Drying conditions at temperatures below 60 °C, the RS content of FDF and ODF50 were higher

than the reported values for GBPF [18, 19, 30] and only lower than the findings of Surendra
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Babu, Mahalakshmi and Parimalavalli [50], who also worked on GBPF, which might be due

to the different variety (Monthan) used in their study. Considering the dramatic increase in the
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levels of digestible starch in both ODF80 and ODF110, it can be suggested that retrogradation
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could be a reason for the disruption of the starch molecule, precisely amylose.
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Figure 2. Starch analysis of green banana flour and wheat flour

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As shown in Fig. 3, the amount of amylose was decreased dramatically by the application of

heat treatment. Contrarily, during the freeze-drying process the amylose crystallites remained
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nearer to the double amount compared to other GBF samples and WF. The linear structure of

amylose (α-1→4) is responsible for the RS2 structure, whereas amylopectin is branched and

its chains are connected through (α-1→6) bonds to form a composite structure [51]. Multiple

significant correlations were found between amylose and RS (r > 0.91, P-value < 0.01) as well

as TS (r > 0.89, p = 0.05), and WHC (r > 0.90 p = 0.05) According to Eshak [16], higher amount

of amylose led to higher a OHC, which was seen in the current study. This, brings about the
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possibility of amylose-lipid complex that has been shown is resistant against α-amylase (RS5).

The FDF results were higher than those found in GBPF reported by Bi, Zhang, Jiang, Hong,

Gu, Cheng, Li and Li [49] but almost the same for ODF50 [31, 52] and even more than culled

plantain starch [24]. As the amount of RS found in FDF and OFD50 were higher than other

samples, it may be advisable for nutritional purposes to use RS as a functional ingredient, as it

contributes to the prevention the colorectal cancer [51, 53].

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3.4. Relative Crystallinity

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The XRD patterns of starches are categorized into three types: A, B and C. A-type starches

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possess surface pores and channels which are slowly digestible, while C-type, can be easily

lost in foods due to the hydrothermal procedures such as boiling, cooking and baking which
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brings about complete or partial starch gelatinization [54]. However, B-type starches do not

have pores in their structure and are inherently resistant to digestion [55]. In green banana, the
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abundance of B-type makes it valuable in terms of health benefits [52].

The XRD patterns of GBF different samples and WF are shown in Fig. 4. Three prominent
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peaks were detected by diffractogram within the diffraction angles of 15° and 25°, which relates
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to B-type crystallinity [55]. While WF showed the least amount of crystallinity, the increased
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drying temperature resulted in the disruption of the starch crystalline section. FDF had 58.34
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% crystallinity, which was the greatest amongst the GBF samples (P < 0.05). These results

explained the existing significant correlation between the obtained RS amount and the effect
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of temperature (r > 0.91, p = 0.05). These results also confirmed that destroying the granular

crystalline structure of starch was associated with decreasing in the amount of RS. This relation

was presented by Zhang and Hamaker [56] who investigated the starch properties in green

banana. The current findings suggest a greater crystallinity in FDF compared to some studies

on GBPF obtained from different cultivars [49]. Furthermore, Wang, Huang and Chen [11]
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reported the effect of cultivar on the percentage of crystallinity in this order: Pisan Awak >

Bluggoe > Cavendish > Pisang.

Figure 3. Amylose and Amylopectin content of green banana flour and wheat flour

Most of the RS found in GBF is type two [4]. The crystalline area is structurally tight and

orderly in a way that makes it hard for it to be eroded by acids and enzymes, whereas the non-

crystalline area (amorphous part) can be easily degraded [27, 28]. In this study, all heat-treated

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flours showed that crystallinity decreased along with the amylose content.

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Figure 1. X-Ray diffraction patterns of banana flours and wheat flour

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3.5. Thermal properties

Thermal properties of the whole GBF and WF are presented in Table 6. All of the thermal
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variables, except for gelatinization range, were significantly higher in GBF samples than the

control (P-value < 0.05). Due to the presence of RS in the GBF samples, a higher Tp than WF
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was found, ranging from 74.99 °C to 75.9 °C. This significant difference could be related to

the high amount of DF in pulp, and especially peel [4]. The values of Tp found in this study
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were higher than those found in corn starch (67.88 °C) [57], potato (64.48 °C) [58] and cassava
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(71.08 °C) [59]. This trend was similar in T0. Almost the same results were reported in previous
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studies with different banana varieties but same drying conditions [60, 61], except for Musa
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paradisiaca Macho, which showed a Tp value of 77 °C [62]; and the freeze-drying method

exhibited a higher Tp than other methods such as ultrasound treatment [20] and pulsed-fluidized
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bed agglomeration [28].

The required energy for breaking the molecular interactions within the starch structure during

gelatinization defines the ∆H [62]. Lower values of ∆H would indicate either a fractional

melting of amylopectin branches or a possible variance in the crystals’ stability related to the

size of the starch granules [61]. For that reason, the control samples had the least amount of

∆H (5.41 J/g), while FDF and ODF50 showed a dramatically higher temperature amongst GBF
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samples. Also, as seen in Table 6, the R value of WF was the highest (11.95 °C), while ODF50

represented the smallest gelatinization range amongst all GBF samples. This can be explained

by the presence of the large amount of amylose per unit mass in flour.

Compared to the results acquired from the oven drying for producing GBPF and GBPeF [58,

63], the freeze-dried sample and ODF50 presented higher values which means that these flours

can potentially be useful in products which delayed pasting is favorable. Also, the current

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results suggest that the incorporation of peel flour into pulp flour may increase the

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gelatinization temperature positively.

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Table 6. Thermal properties of green banana flour samples and wheat flour

4. Conclusion
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Considering that the largest banana producers are developing countries and that thermal

processing is the most common preservation treatment, optimizing a feasible drying method
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that has least negative effects on green banana components, especially its RS2 content, is very

useful for streamlining production of banana flour. Further, banana peel is rich in dietary fibers,
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minerals and RS2 and the production of whole green banana flour was made from whole green
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banana can add nutritional and economic incentives to process. The present study report for
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the first time, investigated the effects of drying conditions on the properties of whole green
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banana flour.

Color analysis showed that FDF had a bright greenish-like flour with the closest browning
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index to wheat flour and the ODF50 treatment may contribute to a better consumer acceptancy

amongst heat-treated green banana samples. The emulsifying properties showed that FDF and

ODF50 can act as an emulsifier in oil-containing food products and they are better than ODF80

and ODF110 samples. Additionally, the high WHC of all GBF samples, especially the FDF,

suggests that they can be applied in food products as a thickening agent.

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High drying temperatures decreased the percentage of RS starch and increased the percentage

of digestible starch. These findings, which were in accordance to high crystallinity and

gelatinization temperature of starch, showed that 50 °C is the best temperature for hot-air oven

drying that can be applied with the least possible effect on RS content. Further, the ODF50

samples were the closest properties to the non-heat-treated sample (i.e. FDF). Future studies

should focus on the particle size distribution, starch granule microstructure and rheological

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properties of the whole green banana flour to understand their potential behavior during

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packaging, transportation and use in food systems.

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Conflict of interest

The authors declare that they do not have any conflict of interest.
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Acknowledgment

The authors would like to express their gratitude to the University of Otago for their support
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of this study.
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Figure 1. Flow chart of green banana flour production by two different drying methods
* WF, wheat flour, FDF, freeze dried flour, ODF50, oven-dried flour at 50 °C, ODF80, oven-dried flour at 80 °C,
ODF110, oven-dried flour at 110 °C.
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Figure 2. Starch analysis of green banana flour and wheat flour
* WF, wheat flour, FDF, freeze dried flour, ODF50, oven-dried flour at 50 °C, ODF80, oven-dried flour at 80 °C,
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ODF110, oven-dried flour at 110 °C.
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Figure 3. Amylose and Amylopectin content of green banana flour and wheat flour
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* WF, wheat flour, FDF, freeze dried flour, ODF50, oven-dried flour at 50 °C, ODF80, oven-dried flour
at 80 °C, ODF110, oven-dried flour at 110 °C.
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20000 Sample Crystallinity SEM

e
18000 WF 41.16 1.23
a
16000 FDF 58.34 1.34

14000 ODF50 48.39b 1.19

ODF80 45.64c 1.41
12000
d
Intensity

ODF110 43.37 1.49

10000

8000
ODF110
6000 ODF80

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ODF50
4000
FDF
2000 WF

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0
5 8 12 15 18 22 25 28 31 35

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2θ (degrees)
Figure 4. X-Ray diffraction patterns of banana flours and wheat flour
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* WF, wheat flour, FDF, freeze dried flour, ODF50, oven-dried flour at 50 °C, ODF80,
oven-dried flour at 80 °C, ODF110, oven-dried flour at 110 °C.
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Table 1. Physicochemical properties and firmness of green banana compared to previous

findings

Data from literature*

Banana characteristics First ripening Second ripening Current findings
stage stage
pH 5.3 ± 0.1 4.8 ± 0.1 5.3 ± 0.1
Soluble solids (°Brix) 5±2 7±1 4.2 ± 1.0
Total solids (g/100 g) NR NR 30.7±2.3
Firmness (N) 26 ± 6 14 ± 2 26.8 ± 3.0

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Titratable acidity 0.24 ± 0.03 0.44 ± 0.03 0.32 ± 0.07
Mean values ± SD of triplicate determinations. NR, not reported

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*
Tribess, Hernández-Uribe, Méndez-Montealvo, Menezes, Bello-Perez and Tadini [28]

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Table 2. The composition of green banana flour and wheat flour samples (db g/100 g)

Moisture Water
Sample Ash Protein Lipid Carbohydrate
Content Activity
WF 9.47a 0.64a 2.83a 9.78a 1.24a 76.68b
FDF 5.27b 0.24c 5.21bc 3.97b 0.92b 84.61a
ODF50 5.09bc 0.25c 5.19b 4.17b 0.93b 84.62a
ODF80 4.56d 0.34bc 5.21bc 4.13b 0.93b 85.14a
ODF110 4.46d 0.39b 5.24bc 4.14b 0.93b 85.26a

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SEM 0.12 0.23 0.61 0.13 0.26 0.71
Mean values of triplicate determinations. Mean values in the same column followed by different letters are
significantly different (P-value < 0.05). SEM, Standard Error of Mean. WF, wheat flour, FDF, freeze dried flour,

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ODF50, oven-dried flour at 50 °C, ODF80, oven-dried flour at 80 °C, ODF110, oven-dried flour at 110 °C.

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Table 3. Color analyses of green banana flour and wheat flour

Browning
Sample L* a* b* Chroma
Index
WF 66.69e 0.87b 11.82a 20.05a 11.85a
FDF 59.72d -0.75a 15.55d 28.45b 15.57e
ODF50 51.46c 2.08c 14.32c 33.67c 14.47d
ODF80 46.55b 2.55d 12.13b 34.93cd 12.40b
ODF110 42.42a 2.97de 13.63c 42.90d 13.94c
SEM 0.76 0.43 0.63 0.42 0.53

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Mean values of triplicate determinations. Mean values in the same column followed by different letters are
significantly different (p≤0.05). SEM, Standard Error of Mean. WF, wheat flour, FDF, freeze dried flour, ODF50,
oven-dried flour at 50 °C, ODF80, oven-dried flour at 80 °C, ODF110, oven-dried flour at 110 °C.

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Table 4. Hydration properties of Green banana flour and wheat flour

T WF FDF ODF50 ODF80 ODF110 SEM

c c a ba a
WHC40 1.97 3.58 3.01 3.00 3.06 0.74
WHC60 4.71b 5.23b 4.31cb 4.86dc 3.90c 0.63
WHC80 7.87ba 9.62a 7.61b 7.64c 7.07c 0.71
OHC40 2.74cb 5.81b 2.77cb 2.70dc 2.49c 0.68
OHC60 2.54c 5.91b 2.77cb 2.76dc 2.53c 0.87

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OHC80 2.34c 5.9a 2.92a 2.74dc 2.47c 0.62
SI40 0.06c 0.12c 0.14b 0.19cb 0.18ba 0.54

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SI60 0.08c 0.13c 0.14b 0.19cb 0.18ba 0.75
SI80 0.16c 0.14d 0.17cb 0.20b 0.20a 0.78

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SP40 3.28c 5.00cb 4.64b 4.65c 4.58b 0.61
SP60 6.24cb 5.96d 6.02cb 6.77b 5.70b 0.88
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SP80 10.73b 11.75b 10.31cb 10.63c 9.93c 0.70
Mean values of triplicate determinations. Mean values in the same line followed by different letters are
significantly different (P-value < 0.05). SEM, Standard Error of Mean. WHC, water holding capacity. OHC, oil
holding capacity. SI, solubility index. SP, swelling power. WF, wheat flour, FDF, freeze dried flour, ODF50,
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Table 5. Emulsifying properties of green banana flour and wheat flour

Sample Emulsion Activity Emulsion Stability

WF 32.61b 9.69e
FDF 32.59b 28.99a
ODF50 36.47a 19.21d
ODF80 32.65b 23.15c
ODF110 30.54c 25.36b
SEM 0.17 0.16
Mean values of triplicate determinations. Mean values in the same column followed by different letters are

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significantly different (P-value < 0.05). SEM, Standard Error of Mean. WF, wheat flour, FDF, freeze dried flour,
ODF50, oven-dried flour at 50 °C, ODF80, oven-dried flour at 80 °C, ODF110, oven-dried flour at 110 °C.

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Table 6. Thermal properties of green banana flour samples and wheat flour

Start
Sample Onset (°C) Maximum (°C) Stop (°C) Delta H (J/g) R (°C)
(°C)
WF 54.7d 56.66d 62.62c 69.42d 5.41e 11.95a
FDF 64.18c 71.08c 74.99b 90.64a 23.16a 8.02b
ODF50 67.73ab 73.45a 75.9a 85.51a 19.48b 4.69d
ODF80 66.69b 71.03c 74.60b 81.61c 10.31d 6.88c
ODF110 68.54a 72.9b 75.86a 84.37b 11.7c 5.92cd
SEM 0.63 0.75 0.85 0.61 0.81 0.59

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Mean values of triplicate determinations. Mean values in the same column followed by different letters are
significantly different (P-value < 0.05). SEM, Standard Error of Mean. WF, wheat flour, FDF, freeze dried flour,
ODF50, oven-dried flour at 50 °C, ODF80, oven-dried flour at 80 °C, ODF110, oven-dried flour at 110 °C.

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