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Article history: Banana resistant starch samples were extracted and isolated from two banana cultivars (Musa AAA group,
Received 5 September 2013 Cavendish subgroup and Musa ABB group, Pisang Awak subgroup) at seven ripening stages during post-
Received in revised form 15 January 2014 harvest storage. The structures of the resistant starch samples were analysed by light microscopy, polar-
Accepted 3 February 2014
ising microscopy, scanning electron microscopy, X-ray diffraction, and infrared spectroscopy.
Available online 12 February 2014
Physicochemical properties (e.g., water-holding capacity, solubility, swelling power, transparency,
starchiodine absorption spectrum, and Brabender microviscoamylograph prole) were determined.
Keywords:
The results revealed signicant differences in microstructure and physicochemical characteristics among
Banana resistant starch
Physicochemical properties
the banana resistant starch samples during different ripening stages. The results of this study provide
Ripening stage valuable information for the potential applications of banana resistant starches.
Starch structure 2014 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2014.02.012
0308-8146/ 2014 Elsevier Ltd. All rights reserved.
320 J. Wang et al. / Food Chemistry 156 (2014) 319325
Kingdom; Soltani et al., 2011), the fruits were divided into seven where m0 is the weight of the starch sample, m1 is the weight of the
ripening states: 1-entirely green; 2-green with a trace of yellow; centrifuge tube, and m2 is the weight of the starch sample and cen-
3-more green than yellow; 4-more yellow than green; 5-yellow trifuge tube following water drainage.
with a trace of green; 6-entirely yellow; 7-entirely yellow with
brown speckles. Bananas at ripening stage 1 were selected and 2.4.2. Solubility and swelling power
purchased from a local market. Subsets of these bananas were Solubility (S) and swelling power (SP) were determined, using
stored at room temperature for different periods of time to attain the method reported by Aparicio-Saguiln et al. (2005). In this
ripening stages 27. However, only bananas at stages 15 were experiment, 20 ml of starch suspension (5 g/100 ml) were trans-
used in this study because RS samples from bananas at stages 6 ferred to centrifuge tubes and heated in a water bath for 30 min
and 7 were colloid-like substances that were difcult to grind. at 50 C, 70 C, or 90 C. After the tubes had cooled to room temper-
All chemicals used in this study were of analytical grade. ature, they were centrifuged at 3000 rpm for 15 min. The sediment
and supernatant were separated; the sediment was dried and
2.2. Isolation and determination of banana resistant starch (BRS) weighed. S and SP were calculated using Eqs. (2) and (3),
content respectively.
A
Using the method reported by Cheng Yanfeng et al. (2008), ba- S % 100% 2
W
nanas were peeled, pulped, and digested with pectinase and amy-
lase to remove pectin, cellulose, protein, and digestible starch. The D
SP % 100% 3
digested banana pulp was centrifuged at 3 000 rpm for 15 min; the W1 S
resulting precipitate was dehydrated at 50 C, ground, and stored
at 5 C. RS content was determined by the method reported by where A is the weight of dry dissolved solids in the supernatant, W
Goni, Garcia-Diz, Manas, & Saura-Calixto (1996). Briey, the meth- is the weight of the sample, and D is the weight of the sediment.
od consisted of the removal of protein and digestible starch, the
solubilisation and enzymatic hydrolysis of RS, and the quantica- 2.4.3. Transparency
tion of RS. Human gastric and intestinal conditions (pH and transit An aqueous starch solution was preparing by mixing 1.0 g of
time) were simulated. starch with 99.0 g of water. This solution was heated in a boiling
water bath for 15 min under continuous stirring and subsequently
cooled to room temperature. The transparency of the resulting
2.3. Structural observations of BRS
starch paste was detected at 620 nm (UV-1800 spectrophotometer,
Shimadzu Co., Japan). Distilled water was used as a blank control,
2.3.1. Light microscopy and polarising microscopy
which was considered to have a transparency of 100%.
BRS samples were dissolved in glycerol (50% concentration) and
observed under a microscope (Vanox BHS-2, Olympus Corporation,
2.4.4. Starchiodine absorption spectra
Japan) using both natural and polarised light.
Spectra of iodine-bound starch samples were determined, using
the method reported by Klucinec and Thompson (1998). BRS
2.3.2. Scanning electron microscopy (SEM) (50 mg) was dispersed into 10.0 ml of DMSO containing 10% of
Particles of BRS powder were scanned, using a S3700N scanning 6.0 M urea. Subsequently, 2.0 ml of the dispersed solution, 25 ml
electron microscope (Hitachi, Japan). Samples were xed on an of distilled water, and 1.0 ml of I2-KI (2.0 mg I2/ml and 20.0 mg
objective table coated with platinum (1020 nm thickness). KI/ml) were pipetted into a 50 ml volumetric ask and mixed.
The mixed solution was brought to a volume of 50 ml with distilled
2.3.3. X-ray diffraction (XRD) water. Control solutions were prepared without BRS. A UVvisible
Cu Ka radiation was used to scan BRS samples over the 2h = 4 spectrophotometer (UV-1800, Shimadzu Co., Japan) was used to
60 range, with a step interval of 0.04, a scanning rate of 17.7 s per scan each sample from 500 to 800 nm; kmax for each sample was
step, a voltage of 40 kV, and a current of 40 mA. The D8 ADVANCE dened as the wavelength that resulted in the highest absorbance
X-ray diffractometer from Bruker Corporation (Germany) was used value.
for the XRD analyses.
2.4.5. Pasting properties
2.3.4. Infrared spectroscopy A microviscoamylograph (Visgraph-E, Brabender Instruments,
BRS samples were pressed in KBr. An infrared spectrometer Inc., Germany) was used to determine the viscosity proles (in
(VECTOR33, Bruker Corporation, Germany) was used to scan the Brabender units, BU) of the starch samples. Dispersions of BRS
samples from 4000 cm1 to 400 cm1 of the infrared region. (6%, dry basis) were transferred to the microviscoamylograph
and subjected to thorough agitation. The dispersion was brought
2.4. Physicochemical properties of BRS to an initial temperature of 30 C and subsequently to 95 C at a
rate of 1.5 C/min. The temperature of the dispersion was main-
2.4.1. Water-holding capacity (WHC) tained at 95 C for 30 min; subsequently, the dispersion was cooled
WHC was measured by the method reported by Toyokawa, to 50 C at a rate of 1.5 C/min and maintained at 50 C for 30 min
Rubenthaler, Powers, and Schanus (1989). Briey, 20 ml of starch (Aparicio-Saguiln et al., 2005).
suspension (5 g/100 ml) were transferred to centrifuge tubes and
heated in a water bath for 15 min at 50 C, 70 C, or 90 C. The 2.5. Statistical analyses
tubes were centrifuged at 3,000 rpm for 15 min. The supernatant
was discarded; tubes containing sediment were placed at a 45 an- Data were analysed by the SPSS statistical software package,
gle for 10 min to allow water drainage and weighed. WHC was cal- v19.0 (IBM company). Data were expressed as means standard
culated by Eq. (1). deviation. One-way analysis of variance (ANOVA) was used to
compare the different BRS samples, Levenes test was used to as-
m2 m1 m0 sess homogeneity of variances, and the Bonferroni test was used
WHC % 100% 1
m0 for multiple comparisons. Statistical signicance was set at
J. Wang et al. / Food Chemistry 156 (2014) 319325 321
P < 0.05. Values followed by the same letter in the same row or col- RS decreased slowly from ripening stages 45; however, the differ-
umn are not signicantly different (P < 0.05) in the tables. ences were signicant. Crystallinity, which represents the degree
of structural order, has signicant effects on hardness, density,
3. Results and discussion transparency, and diffusion (Oxford dictionary of science., 1999).
In the two banana cultivars, RS crystallinity was not reduced at
3.1. Changes in BRS content during ripening the same rate, suggesting that the crystallization behaviour in
starches is possibly different among banana varieties.
BRS content gradually decreased during storage. Cavendish BRS
content decreased rapidly during the rst four ripening stages but 3.2.4. Infrared spectroscopy
decreased slowly during the nal three ripening stages. In contrast, Similar IR spectra of RS were obtained at all ve ripening stages
Pisang Awak BRS content decreased slowly during the initial three for Cavendish and Pisang Awak, indicating that the characteristic
ripening stages but decreased rapidly during the nal four ripening functional groups of RS were not affected by the maturation
stages. At the same ripening stage, Pisang Awak bananas consis- process (Fig. 2). The broad band at 3400 cm1 is caused by OH
tently had higher BRS content than had Cavendish bananas. There groups, whereas the band at 2800 cm1 is generated by CH2 groups
were signicant differences in BRS content between the two bana- (Garcia-Rosas et al., 2009). The peak at 1600 cm1 results from
na cultivars, a result that may be attributed to differences in enzy- carboxylate ion (COO) stretching vibrations in carboxylate
matic reactions that convert starches into sugars. The reaction rate groups. The IR bands at 1300 cm1 and 1022 cm1 are produced
of Cavendish bananas was possibly faster, thereby contributing to a by COH bending and COH bending vibrations, respectively.
rapid reduction in BRS content. The skeletal modes of the pyranose ring generate the peak at
620 cm1.
1 2 3 4 5
(a) Optical microscopy images (200) of Musa AAA Cavendish BRS
1 2 3 4 5
(b) Optical microscopy images (200) of Musa ABB Pisang Awak BRS
1 2 3 4 5
(c) Polarised light microscopy images (200) of Musa AAA Cavendish BRS
1 2 3 4 5
(d) Polarised light microscopy images (200) of Musa ABB Pisang Awak BRS
1 2 3 4 5
(e) Scanning electron microscopy images (500) of Musa AAA Cavendish BRS
1 2 3 4 5
(f) Scanning electron microscopy images (500) of Musa ABB Pisang Awak BRS
Fig. 1. Optical microscopy, polarised light microscopy and scanning electron microscopy images of resistant starch samples isolated from bananas at different ripening
stages. (The numbers 15 indicate the ripening stage of each banana sample.)
and less amylose than does Pisang Awak BRS. The maximum stages 1, 2, 3, 4 and 5, the kmax of Pisang Awak BRS samples oc-
absorption wavelengths of Cavendish BRS at ripening stages 15 curred at 585, 600, 605, 573, and 595 nm, respectively, suggesting
were similar at approximately 570 nm. However, the maximum that the amylopectin content of Pisang Awak BRS decreased
absorption wavelengths of Pisang Awak BRS shifted as bananas quickly prior to ripening stage 3, causing kmax to shift to wave-
matured through ripening stages 15. In particular, for ripening lengths near the maximum absorption wavelength of amylose. It
J. Wang et al. / Food Chemistry 156 (2014) 319325 323
Table 1
The relative degrees of crystallinity (%) of banana resistant starch samples.*
Values followed by the same letter in the same row are not signicantly different (P < 0.05).
*
Mean of three replicates standard error.
Fig. 2. Infrared spectra of resistant starch samples isolated from bananas at different ripening stages. (The numbers 15 indicate the ripening stage of each banana sample.)
Table 2
The water-holding capacity, solubility, swelling power, and transparency of resistant starch samples isolated from bananas at different ripening stages.*
Values followed by the same letter in the same row are not signicantly different (P < 0.05).
*
Mean of three replicates standard error.
is likely that amylose was rapidly degraded between ripening 3.3.5. Pasting properties
stages 3 and 4, causing kmax to shift to 573 nm. At ripening stage The pasting properties of BRS were measured by a Brabender
5, both amylopectin and amylose contents continued to decrease microviscoamylograph; the results are shown in Table 3. The initial
in the Cavendish and Pisang Awak cultivars, as indicated by a gelatinisation temperatures of Cavendish BRS were similar during
reduction in the starchiodine absorption peaks with increasing ripening stages 13, whereas the gelatinisation temperatures of Pi-
banana fruit maturity. sang Awak BRS increased as banana fruits matured. In Cavendish
324 J. Wang et al. / Food Chemistry 156 (2014) 319325
Fig. 3. Starchiodine absorption spectra of resistant starch samples isolated from bananas at different ripening stages. (The numbers 15 indicate the ripening stage of each
banana sample.)
Table 3
Banana resistant starch viscosity parameters, as measured by the Brabender microviscoamylograph .*
Banana variety Ripe stage A(C) B(BU) C(BU) D(BU) E(BU) F(BU) B-D(BU) E-D(BU) E-B(BU)
Musa AAA Cavendish 1 78.8 0.5a 498 35a 490 36a 377 40a 588 33a 519 26a 121 211 90
2 78.4 0.7a 469 31a 467 32a 311 30b 501 35b 445 34b 158 190 32
3 79.1 0.7a 466 23a 462 28a 328 39b 496 31b 442 33b 138 168 30
Musa ABB Pisang Awak 1 65.5 1.0b 302 26b 300 26b 240 35c 329 22c 293 20c 62 89 27
2 79.9 0.8c 270 31b 263 37b 209 24c 292 30c 272 32c 61 83 22
3 80.0 0.7c 266 37b 262 44b 230 40c 345 37c 314 31c 36 115 79
BU = Brabender unit.
Values followed by the same letter in the same column are not signicantly different (P < 0.05).
*
A-initial gelatinization temperature; B-peak viscosity (BU); C-viscosity at 95 C (BU); D-viscosity after 30 min at 95 C (BU); E-viscosity at 50 C (BU); F-viscosity after
30 min at 50 C (BU); B-D: Breakdown; E-D: Consistency. E-B: Setback. Mean of three replicates standard error.
BRS, the properties of peak viscosity after gelatinisation (point B), Awak banana fruits. The Maltese crosses of BRS were clear under
viscosity at 95 C (point C), viscosity at 50 C (point E), and viscos- polarised light at the beginning of storage but became weak during
ity after 30 min at 50 C (point F) decreased during ripening stages the ripening process. BRS crystallinity gradually decreased during
13, whereas the viscosity after 30 min at 95 C (point D) was not the ripening process. The infrared spectra of Cavendish and Pisang
affected. In contrast, in Pisang Awak BRS, only the peak viscosity at Awak BRS were similar and remained relatively unaffected
point B and the viscosity at 95 C (point C) decreased during ripen- throughout the maturation of banana fruits, suggesting that the
ing stages 13. The viscosity of Cavendish BRS was consistently typical functional groups of RS were well maintained at all ve
higher than the viscosity of Pisang Awak BRS at points AF. Caven- examined ripening stages.
dish BRS had the highest peak viscosity (point B) during the rst Differences in WHC, solubility, swelling power, and transpar-
ripening stage. ency were observed in BRS from the two cultivars and at different
The heat stability of BRS paste determines the viscosity after ripening stages. Starchiodine absorption spectra indicated that
30 min of incubation at 95 C (B-D; breakdown), whereas the cold Cavendish BRS contained more amylopectin and less amylose than
stability of BRS determines the viscosity after 30 min of incubation did Pisang Awak BRS. The degradation processes of amylopectin
at 50 C (E-D; consistency). The setback refers to the difference be- and amylose in the two cultivars were asynchronous. Pasting prop-
tween the peak viscosity after gelatinisation and the viscosity at erties, determined by Brabender microviscoamylograph analyses,
50 C (Qian & Kuhn, 1999). In this study, Cavendish BRS had the revealed differences in the heat stability (breakdown value), cold
highest breakdown (B-D) and consistency (E-D) values at ripening stability, and setback characteristics of BRS from the two cultivars
stage 2 and the maximum setback (E-B) value at ripening stage 1. at ripening stages. In general, viscosity values were higher for
Pisang Awak BRS had the highest breakdown value during the rst Cavendish BRS than for Pisang Awak BRS. These results reveal that
ripening stage and the highest consistency and setback values dur- appropriate cultivars and ripening stages should be chosen for the
ing ripening stage 3. Moreover, higher breakdown (B-D), consis- preparation of RS products.
tency (E-D), and setback (E-B) values were obtained from
Cavendish BRS than from Pisang Awak BRS, with the exception of
the setback (E-B) value at the third ripening stage. Acknowledgments
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