BRAZILIAN JOURNAL OF THERMAL ANALYSIS

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Thermal, structural, pasting properties and digestibility investigation on green

banana starch: Combination of organic acids and Heat–Moisture Treatment

Jennifer Santos Ramos1, Bárbara Ruivo Válio Barretti1, Vanessa Soltes de Almeida1, Vivian Cristina Ito1,2, Ivo Mottin

Demiate1, Egon Schnitzler1, Luiz Gustavo Lacerda1*

1 Food Science and Technology Graduate Program, State University of Ponta Grossa (UEPG), Ponta Grossa, Brazil

2 Department of Agroindustry, Food, and Nutrition, “Luiz de Queiroz” College of Agriculture, University of São Paulo

(USP), Piracicaba, Brazil

*Luiz Gustavo Lacerda, e-mail: lglacerda@uepg.br Phone: +55 41 984320018. Food Science and Technology Graduate

Program – State University of Ponta Grossa (UEPG), Av. Carlos Cavalcanti, 4748 Uvaranas Campus, ZIP 84.030-900 –

Ponta Grossa, PR Brazil.

Received: 01/03/2020

Published online: 10/10/2020

ABSTRACT

Green banana is a great potential source for starch. Its starch functional

properties can be improved by applying various innovative and safe

technologies, using either single or double modification. The objective of

this study was to investigate the impacts of combination of organic acids

(lactic and citric) and heat-moisture treatment (HMT) on the thermal,

structural, pasting properties, and digestibility of green banana starch.

After conventional HMT (with deionised water), the transition

temperatures (onset - To, peak - Tp and endset - Tc) increased. However,

the starches treated by HMT using both lactic and citric acid did not

present gelatinisation peaks. The starch samples modified showed a

significant decrease in relative crystallinity. For pasting properties, it can

be seen that there was an increase in pasting temperature and a decrease in

peak viscosity, breakdown, setback, and final viscosity for samples

modified. Combination of organic acids and HMT changed the contents of

the rapidly digestible starch (RDS), slowly digestible starch (SDS) and

resistant starch (RS). It was observed the increase in RDS and SDS content

and a decrease in RS. The RS fraction decreased from 77.63% to 61.26%

when treated with lactic acid in the presence of heat and moisture. These

results can be indicating applications for green banana starch in functional

foods with reduced viscosity.

Keywords: Citric acid, lactic

acid, resistant starch, viscosity.

BRAZILIAN JOURNAL OF THERMAL ANALYSIS

http://www.bjta.com.br

1 Introduction

Glycemic indexes and predisposition to

diabetes have been increasing worldwide based

on research by the International Diabetes

Federation. In 2014, 387 million people

worldwide had diabetes mellitus (DM), aged

between 20 and 79 years. In Brazil, 11.6

million people have diabetes, with an estimate

for 2030 increasing to 16.3 million people.

According to the American Diabetes

Association, DM has been a major public

health problem in the country, becoming a

precursor of risk for the development of

cardiovascular diseases [1]. The importance of

consuming foods with a low glycemic index

consists of less insulin release in the body to try

to keep blood glucose levels low. This leads to

satiety, reducing the risk of developing diseases

such as type 2 diabetes mellitus [2].

The choice of the green banana is due to

its aspects in relation to the nutritional and

sensory value. The banana has an energy and

fiber source and contains vitamins and minerals

[3]. The energy supply of the banana is due to

the presence of starch in high concentrations

(approximately 70-80% on a dry basis) from

the flour. This high content, coupled with the

fact that it presents resistance, arouses interest

as an alternative source of food and use in the

food industry, such as in diet products, baby

foods and bakery products [4,5].

In general, starches in their native form

have technological limitations due to their low

thermal resistance, reduced shear tolerance and

high retrogradation in storage [6]. Thus, to

enhance its industrial use, the biopolymer

undergoes chemical modifications; biological;

physical properties such as HMT or combined.

Thermal modification can alter its

digestibility [5]. After modifications, resistant

starch can cause a reduction in the glycemic

index, increase in satiety, also helping to reduce

the risk of certain diseases such as obesity and

hyperglycemia [2]. In addition, acids, such as

lactic and citric, are food-grade ingredients,

which can be used in the food processing as

inorganic acids replacements. Thus, the present

study investigated the impact of a combination

of organic acids and heat-moisture treatment on

the thermal, structural, pasting properties, and

digestibility, of green banana starch.

2 Materials and methods

2.1 Materials and Methods

For the experiments, green banana flour

was purchased from the local trade in Ponta

Grossa / PR / Brazil. The extraction of starch

from the flour was carried out according to the

methodology of UDA et al. [7]. All reagents

used were of analytical standard. Porcine

pancreatic α-amylase (E.C. 3.2.1.1, 8 x USP

specifications, P7545) and Aspergillus niger

amyloglucosidase (E.C. 3.2.1.3, A7095, ≥

260U/mL) were purchased from Sigma

Chemical Co. (St. Louis, MO, USA). The

remaining reagents were of analytical grade.

2.2 Acid and heat-moisture treatment (HMT)

To carry out the modification by HMT,

the moisture content of the starches was

adjusted by 22% with the addition of organic

acids (lactic and citric) at a concentration of 0.2

mol L-1 and water. The samples were submitted

to controlled heat in an oven of 110 ºC for 8 h.

Then, the samples were neutralized with 1mol

L-1 NaOH and dried in an oven at 40 ºC for 24h

[8]. The samples were identified as follows:

HMT H2O (heat–moisture treatment with

deionised water); HMT LA (heat–moisture

treatment with lactic acid) and HMT CA (heat–

moisture treatment with citric acid).

2.3 Differential scanning calorimetry

Differential Scanning Calorimetry was

performed using DSC-Q200 equipment (TA-

Instruments, USA). The conditions used were

air flow of 50 mL min-1, heating range from 20

to 100 ºC and heating rate of 10 ºC min-1.

Samples were suspended in water and sealed in

aluminum crucibles [9].

2.4 Powder X-ray diffractometry (PXRD)

Powder X-ray diffractometry was

performed using an Ultima 4 X-ray

diffractometer (Rigaku, JAPAN) with CuKα

radiation, in the 40 kV and 20 mA

configurations and with an interval in the

angular range of 3 º to 40 º to 2 (θ) [10].

2.5 Pasting properties (Rapid Visco Analyser -

RVA)

The paste properties were evaluated by

the viscoamylographic profile obtained in the

RVA-4 equipment (Newport Scientific,

Australia). A suspension in water of 8% (w / w)

of dry starch (28 g total weight) was prepared

and subjected to a heating cycle at 95 °C and

controlled cooling under constant circular

stirring. The analysis totaling 23 minutes,

profile consists of continuous heating from 50

to 95 ° C at a heating rate of 6 ° C min-1,

followed by cooling until completion of the

analysis [11].

2.6 In vitro digestibility

In vitro digestibility was determined

according to the Englyst method [12] with

modifications [13]. A 900 mg mass of green

banana starch was suspended in a sodium

acetate buffer solution (0.1 mol L-1, pH 5.2).

Then, the samples were submitted to a shaking

water bath at 37 ºC. Then starch was

hydrolysed using porcine pancreatin extract and

A. niger amyloglucosidase (Sigma-Aldrich Co.,

St. Louis, MO) with continuous agitation (100

movements per minute). The contents of fast

digesting (RDS) and slow (SDS) starch were

measured after incubation periods of 20 and

120 min, respectively, and the fraction that was

not hydrolysed at the end of 120 min was

defined as resistant starch (RS).

2.7 Statistical analysis

The results were expressed as mean ±

standard deviation and were analysed using

Action Stat 3.3 software (Estatcamp, Sao

Paulo, Brazil). Tukey's tests were conducted to

determine the differences between the means at

a 95% confidence level (p < 0.05).

3 Results and discussion

3.1 Differential scanning calorimetry

The DSC curves of samples are

presented in Figure 1. All temperatures (onset –

To, peak – Tp and endset – Tc) and

gelatinisation enthalpies of native and modified

samples of green banana are presented in Table

1.

After conventional HMT (with

deionised water) treatment, the transition

temperatures revealed an increasing. Possibly,

after HMT, there are a stronger interaction

between amylose-amylose chains that can

reduce the mobility of amorphous regions

leading to increase in onset, peak, and

conclusion temperature [14].

Figure. 1 - DSC curves of green banana starch. B

Native – Green banana starch, HMT H2O - distilled

water heat moisture treatment, HMT LA - lactic acid

heat moisture treatment and HMT CA – citric acid heat

moisture tretament.

In accordance to a recent study by

MAIOR et al. [15] treated starch by HMT with

both lactic and citric acid did not present

gelatinisation curves. It is possible to observe

similar behaviour where curves showed lower

pasting properties by RVA analysis (Figure 3).

LIU et al. [16] studied maize starch

modification using HMT with organic acids

and according to them; this combination can

affect the internal structure of starch granules.

HMT with acid treatments can promote the

appearance of structures with different melting

temperatures and damage the gel-forming

ability. The reduction in Hgel observed can be

explained by the degradation of double helices

present mainly in the crystalline region of

starch granules [15].

Table 1 - Results of DSC curves for normal and modified green banana starches. B – green banana

starch; HMT H2O - distilled water heat-moisture treatment, HMT LA - lactic acid heat-moisture

treatment, HMT CA - citric acid heat-moisture treatment.

Treatments To/ °C TP/ °C Tc/ °C ΔHgel/J.g-1

B Native 59.1a±0.60 64.0b±0.10 69.7b±0.20 10.2a±0.90

HMT H2O 60.5a±0.70 72.0a±0.10 80.2a±1.20 5.5b±0.40

HMT LA - - - -

HMT CA - - - -

Note: To – onset, TP - peak temperature, Tc - conclusion temperature, ΔHgel - gelatinization enthalpy.

Values presented as mean values ± standard deviation. The values followed by the same letter in the

same column are not significantly different by Tukey's test (p <0.05).

3.2 Powder X-ray diffractometry (PXRD)

The X-ray diffraction patterns and

relative crystallinities of the green banana

starch samples are given in Figure 2. The native

starch exhibited an XRD pattern characterised

by the presence of a peak at 5°, peaks at 15°

and with an around 17° and a broad peak at 23°

2θ, this diffraction pattern of the banana

starches is consistent with a B-type crystallinity

pattern [17].

On the other hand, when HMT was

employed the diffraction pattern changed, with

intensity decrease until peak in 5°.

Similar results were found in studies

with potato starch treated with HMT and lactic

and citric acids [18]. Due to the thermal energy

and moisture of treatment can occur to the

disruption and reformation of starch chain

interactions within the amorphous and

crystalline domains causing the change in the

XRD pattern [5,6].

Figure. 2 - Diffractograms of green banana starches and

relative crystallinity (RC %) - B Native – Green banana

starch, HMT H2O - distilled water heat moisture

treatment, HMT LA - lactic acid heat moisture treatment

and HMT CA – citric acid heat moisture treatment.

The relative crystallinity of green

banana starch native was approximately

30.18%, which was consistent with other

studies on native green banana starch [5]. The

relative crystallinity of modified starches

decreased to around 24.10%. The high

temperature during HMT can decrease in

relative crystallinity due the disruption of

hydrogen bonds between the double helices

[19].

3.3 Pasting properties (Rapid Visco Analyser -

RVA)

The pasting properties show us its

performance when used in the food industry for

manufacturing pasta, soups, children’s foods,

and frozen products. According to MAIOR et

al. [15] pasting properties are studied to

observe changes in viscosity during the heating

of a starch suspension. The evaluation of these

properties is important when formulating a

product that is going to be exposed to heating

in acidic conditions.

The HMT performed under acid

conditions promoted significant changes in the

starch pasting properties relative to the native

green banana starch. It can be seen that there

was an increase in pasting temperature and a

decrease in peak viscosity, breakdown, setback,

and final viscosity for samples modified. The

pasting profiles are illustrated in Figure 3 and

the values related to the viscoamylographic

analyses obtained by RVA curves are shown in

Table 2.

Figure 3 - RVA profile of green banana starch. B

NATIVE – Green banana starch, HMT H2O - distilled

water heat moisture treatment, HMT LA - lactic acid

heat moisture treatment and HMT CA – citric acid heat

moisture treatment.

The pasting temperature increased in the

green banana starch samples modified. Similar

behaviour was observed by [20] analyzing

Prata green banana starch modified with HMT

(moisture contents of 15%, 20%, and 25%).

This increase suggests that the granules, after

modification, initiated to swell at higher

temperatures than the native green banana

starch.

The peak viscosity, breakdown, setback,

and final viscosity decreased in the green

banana starch samples modified. CAHYANA

et al. [5] observed similar changes in HMT of

the green banana flour. In the research carried

by SILVA et al. [21] with acid modification,

there was also a decrease in the measured

viscosities of the green banana starch.

Table 2 – RVA profile for normal and modified green banana starches. B – green banana starch; HMT H2O -

distilled water heat-moisture treatment, HMT LA - lactic acid heat-moisture treatment, HMT CA - citric acid

heat-moisture treatment.

Treatments Pasting

temperature/ºC

Peak Viscosity

/mPa s

Breakdown

/mPa s

Setback

/mPa s

Final Viscosity

/mPa s

B Native 72.77c ± 0.12 2877.50a ± 0.70 1190.50a ± 0.70 1529.50a ± 2.12 3216.50a ± 2.12

HMT H2O 91.90b± 0.07 1214.00b ± 1.41 - 498.00b ± 1.41 1716.00b± 1.41

HMT LA 94.05a± 0.65 798.00c ± 1.41 0.50c± 1.41 353.00b ± 1.41 1106.00c± 1.41

HMT CA - 471.00d ± 1.41 15.00b ± 1.41 102.00c ± 2.82 558.00d± 2.82

Note: mPa s - millipascal-second, s - second. Values presented as mean values ± standard deviation. Values

followed by the same letter in the same column are not significantly different by Tukey’s test (p < 0.05).

MAIOR et al. [15] also found a reduction

in the measured viscosities when studying

maize starches modified by organic acid and

HMT. JUANSANG et al. [22] reported that the

decreased paste viscosity is caused by

alterations in the amorphous fraction, and these

amorphous fractions may be hydrolysed under

acidic conditions. This behaviour results in the

close packing of starch chains. Therefore, the

combination with HMT can result in the

absorption of less water, in other words, less

viscosity.

After combination of organic acids and

HMT the decrease in breakdown shows that

shear stability improved and the tendency to

starch retrogradation was reduced (setback).

The decrease in setback and final viscosities

mainly occurs due to the reordering or

polymerisation of leached amylose and

amylopectin [23].

3.4 In vitro digestibility

The values of starch fractions rapidly

digestible starch (RDS), slowly digestible

starch (SDS) and resistant starch (RS) are

described in Table 3. Native green banana

starch had lower rapidly digestible (9.72%) and

higher resistant (77.63%) contents compared to

potato (18 and 60%, respectively) [24][24] or

maize (29.77% and 39.11%, respectively)[16]

starches.

Previous studies with green banana

starch corroborate the in vitro digestibility

results obtained. For example, AMAYA et al.

[25] found low content of RDS (4.23), SDS

(10.79) and high content of resistant starch,

about 85%. A smooth, dense surface of native

banana starch granules may explain its

resistance to enzymatic digestion [26].

Table 3 - RDS, SDS and RS fractions of native and

modified green banana starches. B Native – green banana

native starch; HMT H2O - distilled water heat-moisture

treatment, HMT LA - lactic acid heat-moisture treatment,

HMT CA - citric acid heat-moisture treatment

Note: Values presented as mean values ± standard

deviation. The values followed by the same letter in the

same column are not significantly different by Tukey's

test (p <0.05).

Previous studies with green banana

starch corroborate the in vitro digestibility

results obtained. For example, AMAYA et al.

[25] found low content of RDS (4.23), SDS

(10.79) and high content of resistant starch,

about 85%. A smooth, dense surface of native

banana starch granules may explain its

resistance to enzymatic digestion [26].

Modification by HMT changed the

composition of the RDS, SDS and RS contents.

In general, was observed the pronounced

increase in SDS content (Table 3). This

behavior was also observed in sweet potato and

yam starches when treated with HMT and

organic acids [8]. The cross-linked and

esterification modification also caused an

increase in SDS in banana starches [25].

The RS content of the green banana

starch decreased significantly (p ≤ 0.05) after

acid and acid-heat moisture treatment. The RS

fraction decreased from 77.63% to 61.26%

when treated with lactic acid after treated by

heat moisture. The modification with citric acid

was the mildest in relation to the decrease in

RS levels. On the other hand, in corn and

sorghum starches treated with HMT and citric

and lactic acids the contents were significantly

increased [27].

Changes in the behaviour of green

banana starches during digestibility may be

related to changes in the X-ray diffraction

pattern. According to the literature have

demonstrated the correlation of the crystalline

type or polymorph with its digestibility [5].

According CAHYANA et al. [5] HMT

can increase the porosity of granules with

several holes, providing more accessibility,

thus increasing digestibility. The HMT

treatment combined with organic acids may

have intensified this accessibility of the

enzymes to the granules as indicated by the

lower RS and increased SDS and RDS

contents.

Conclusions

Modification of the green banana starch

by the organic acids with heat–moisture

treatment provided significat changes in the

thermal, structural, pasting properties, and

digestibility. In the thermal analysis, the

Treatments RDS % SDS % RS %

B Native 9.72c ± 0.20 12.66c ± 0.45 77.63a ± 0.65

HMT H2O 13.32b ± 0.07 21.37a ± 0.18 65.30c ±0.18

HMT LA 16.54a ± 0.08 22.22a ± 0.08 61.26d±0.18

HMT CA 12.82c ± 0.17 18.62b ± 0.32 68.55b ±0.15

transition temperatures revealed an increasing,

due to a stronger interaction between amylose-

amylose chains that reduced the mobility of

amorphous regions. The diffraction pattern

changed, with intensity decrease until peak in

5°. The starch samples modified showed a

significant decrease in relative crystallinity.

It can be seen that there was an increase

in pasting temperature and a decrease in peak

viscosity, breakdown, setback, and final

viscosity for samples modified. The decrease in

breakdown shows that shear stability improved

and the tendency to starch retrogradation was

reduced (setback).

In digestibility, there are changed the

composition of the rapidly digestible starch

(RDS), slowly digestible starch (SDS) and

resistant starch (RS) contents. It was observed

the increase in RDS and SDS content and a

decrease in RS. The RS fraction decreased

from 77.63% to 61.26% when treated with

lactic acid in the presence of heat and moisture.

Our results provide information on the

combination of organic acids and heat–

moisture treatment, in green banana starch. The

lower viscosities of the studied samples suggest

that green banana starch modified could be use

in children’s foods, for example. However,

more starch studies should be performed in

order to improve its technological and

nutritional applications.

Acknowledgements

The authors are grateful to the

Coordination for the Improvement of Personnel

in Higher Level (CAPES), and CNPq (Process

155859/2018-8 and 307654/2017-6), for

scholarship and C-LABMU-UEPG for the

infrastructure. E. Schnitzler, I. M. Demiate, and

L. G. Lacerda are research fellows from CNPq.

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Resumo (em português)

A banana verde é uma potencial fonte de amido. As propriedades funcionais dos grânulos de amido

podem ser aprimoradas utilizando-se diversas tecnologias inovadoras e seguras utilizando-se para tanto

modificações simples ou duplas, por exemplo. O presente trabalho teve por objetivo investigar os

impactos da combinação de ácidos orgânicos (lático e cítrico) combinados com tratamento de umidade

e calor (HMT) nas propriedades térmicas, estruturais, de pasta e digestibilidade do amido de banana

verde. Após o tratamento convencional com HMT (com água), as temperaturas de transição

aumentaram. No entanto, os amidos tratados por HMT usando ácido lático e cítrico não apresentaram

picos de gelatinização. As amostras de amido modificadas mostraram uma diminuição significativa na

cristalinidade relativa. Para propriedades de pasta, foi observado que houve um aumento na

temperatura de pasta e uma diminuição no pico de viscosidade, quebra, setback e viscosidade final das

amostras modificadas. A combinação de ácidos orgânicos e HMT alterou a composição dos teores de

amido rapidamente digerível (RDS), amido lentamente digerível (SDS) e amido resistente (RS).

Observou-se aumento do conteúdo de RDS e SDS e diminuição do RS. A fração RS diminuiu de

77,63% para 61,26% quando tratada com ácido lático no tratamento de HMT. Esses resultados podem

indicar aplicações de amido de banana verde em alimentos funcionais com viscosidade reduzida.