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Physicochemical, thermal, and pasting

properties of flours and starches of eight
Brazilian maize landraces (Zea mays L.)
ARTICLE in FOOD HYDROCOLLOIDS MARCH 2013
Impact Factor: 4.09 DOI: 10.1016/j.foodhyd.2012.08.005

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Food Hydrocolloids 30 (2013) 614e624

Contents lists available at SciVerse ScienceDirect

Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Physicochemical, thermal, and pasting properties of ours and starches of eight

Brazilian maize landraces (Zea mays L.)
Virgilio Gavicho Uarrota a, *, Edna Regina Amante b, Ivo Mottin Demiate c, Flavia Vieira d,
Ivonne Delgadillo d, Marcelo Maraschin a
a

Plant Morphogenesis and Biochemistry Laboratory, Federal University of Santa Catarina, 1346, SC 401 Road, P.O. Box 476, SC Florianopolis, Brazil
Laboratory of Fruits and Vegetables, Departament of Food Science and Technology, Federal University of Santa Catarina, Florianopolis, Brazil
Department of Food Engineering, University of Ponta Grossa, Paran, Brazil
d
Department of Chemistry, University of Aveiro, Portugal
b
c

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 5 December 2011
Accepted 7 August 2012

Both genetic and environmental factors create signicant variation in the amount and quality of maize
landrace constituents. Details on the ours and starch characteristics have not been fully investigated.
The physicochemical, pasting and thermal properties of 8 promising cultivars were assessed in this study
and those properties were correlated. Higher values of swelling and solubility (RJ e 13.14%; 14.39%), lipid
content (MG e 5.53%), WBC (PR e 18.89%), and amylose content (PR e 27.43%) were found for those
genotypes. Lower onset temperatures of gelatinization (To) were observed for RX-F1 (66.1
C) as RX-F1
(68.7
C) genotype showed the lower pasting temperatures. A wide range of viscosity values was
found among the maize landraces (MG-F0, 343 mPa s and RJ-F1, 175 mPa s) as well as for the retrogradation (R8C-F1, 796 mPa s and RX-F1, 22 mPa s). ATR-FTIR spectroscopy revealed amylose, amylopectin, lipids, and proteins as major ours constituents and their differences were discriminated by PCA
analysis.
2012 Elsevier Ltd. All rights reserved.

Keywords:
Maize landraces (Zea mays L.)
Chemometrics
Starch
Amylose
Viscosity
Solubility
ATReFTIR

1. Introduction
Recent advances in biotechnology have accelerated the development and characterization of new crop genotypes and given rise
some questions related to food security, conservation of plant
genetic resources, access and sustainable use of biological diversity,
and environmental friendly agricultural production models. Indeed,
a signicant part of agro-systems worldwide have moved from
subsistence to intensive and market-oriented cultivation systems
which commonly cultivate genetically improved varieties. Accordingly, local and creole genotypes (landraces e Zaid, Hughes,
Porceddu, & Nicholas, 2001) have continuously been replaced in
cultivation systems worldwide, leading to a reduction of the genetic
diversity of important crop species such as maize (Zea mays L. e
Lemos & Maraschin, 2008). This can result in an increased vulnerability of that species to pest and diseases, restricting future
adaptive potential and uses of it (FAO, 1998, p. 510; Zaid et al., 2001).
In fact, it is well known that the main reasons given for loss of agrobiodiversity (e.g., maize landraces e ML) are the replacement of
local varieties by modern varieties, market integration and

* Corresponding author. Tel.: 5548 96128360; fax: 5548 37215400.

E-mail address: uaceleste@yahoo.com.br (V.G. Uarrota).
0268-005X/$ e see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodhyd.2012.08.005

industrialization of agriculture including agro-processing and

marketing requirements (Drucker, Costa, & Magnusson, 2008).
In South hemisphere countries smallholder farmers have cultivated hybrid Z. mays varieties to provide raw materials for agroindustry and, in parallel, maintain agro-ecological production
systems of ML which have a current and meaningful local use in
human and animal nutrition. Such genotypes have been selected
over generation for stability under low-external input conditions
and adverse environments and generally could not compete with
new high-yielding varieties bred for intensive production systems
(FAO, 1998, p. 510). Besides, ML generally show distinct characteristics as to the standard established by the market, specially
regarding their agronomic traits and continuous offering of raw
material (e.g., grains and ours), with difculties to entering the
commercial circuit. This weak commerce appeal makes difcult
their cultivation as some initiatives are needed to support small
farmers interested in keeping them under crop. On the other hand,
at present there is a paradox characterized by the food standardization of one side, with over 75% of human food being based in less
than a ten species, and on the other side the elevation of the
demand for more diversied, natural, healthful and organically
(syn. biologically) foods such as ML ours (Vogt, 2005, 116 p.).
In this context, the creation of strategies to maintain landraces
on cultivation is crucial and one possible approach for that involves

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624

the increase of smallholder farmers incomes. Thus, a suitable

approach seems to be the characterization of the landraces raw
materials regarding their chemical traits that industrial applications might be envisaged and developed. Such an approach is
thought to be an add-value alternative to increase the economic
signicance of those genotypes, directly contributing for the
improvement of the family agriculture sector and preserving that
important germplasm. Indeed, some landraces have been used for
the development of new commercial varieties with desired characteristics that may result in innovations on food, chemical, pharmaceutical, and personal care industries.
Maize ours typically present a high starch and a wide range of
food and non-food applications. Starches, because of their desirable
physical and chemical properties, are known to be suitable for use as
thickeners, extenders, stabilizers, gelling agents, dietary calories,
and texture modiers in food formulations (Adebooye & Singh,
2008). To enhance the suitability of different products, starch is
modied using chemical modications techniques, e.g., hydroxypropylation, acetylation and cross-linking (Kaur, Singh, & Singh,
2004). However, the use of chemically modied starches in food
products is either inhibited or discouraged in a number of countries.
Starches are found as water-insoluble granules of varying sizes
and characteristic shapes depending on their botanical source. They
are composed by a linear a-(1 / 4)-glucan, amylose (AM) and an a(1 / 6)-branched a-(1 / 4)-glucan, amylopectin (AP). Such starch
components differ in their secondary and tertiary structures, giving
rise to distinct arrangements of the crystalline and amorphous
areas in the granules, which impart variability to the chemical and
physical properties of that polysaccharide (Mukerjea & Robyt,
2010).
Amylose is used in the textile industry as a sizing and shing
agent, in the food industry as a thickening, stabilizing, gelling,
encapsulating agent and in the paper industry for adhesives,
binding agents and surface sizing applications (Young, 1984).
Common amylose sources are inappropriate to food industry
purposes because of the chemical residues left by fractionation
methods. To overcome this constraint, high-amylose maize genotypes (up to 70% amylose) have been developed. On the other hand,
the branch chain length of amylopectin has been related to affect
starch crystallization (Garcia, Martino, & Zaritzky, 1995), gelatinization and pasting properties (Singh, Inouchi, & Nishinari, 2005).
Studies on the functional properties of the starch have been
carried out mostly using hybrid varieties of maize, as reports on the
physicochemical properties of ours and starches of maize landraces are scarce. Such genotypes are claimed to show high genetic
variability, a trait that might be correlated in any extension to
a hypothesized chemical diversity, i.e., distinct types of starches
with interesting possibilities for industrial applications. Besides, in
several cases, the focus of investigation on maize has been on starch
gel viscoelasticity, while details on the ours (of whole grain or
decorticated grain) and starch characteristics have not been fully
investigated, especially for maize landraces. This study was
designed to investigate the physicochemical and thermal properties of eight Brazilian ML, focusing on their ours and starchy
fractions. The results are expected to be useful in providing guidance on the potential for industrial uses of those biomasses,
improving the knowledge on the ML biomasses and stimulating the
production and conservation of such important genetic resource.
2. Material and methods
2.1. Selection of maize landraces
Maize landrace grains were produced (2008/2009 harvest)
under agro-ecological management by small farmers at the far-

615

western region of Santa Catarina State, southern Brazil (Anchieta

county, 26
3101100 S, 53
200 2600 W). Eight genotypes were selected
for this study according to their importance for local populations as
follows: Roxo (RX), Palha-Roxa (PR), Mato Grosso Palha Roxa (MG),
Rajado (RJ), Rajado 8 Carreiras (R8C), Roxo do Emilio (RXE), MPA1
(refer to a local maize population-composite variety, grown by
farmers at Anchieta county based on Brazilian MPAeMovimento de
Pequenos Agricultores) and Lngua de Papagaio (LP). For purpose of
comparison as to the physicochemical characteristics in study, two
maize hybrid varieties, e.g., BR SC 154 and Fortuna, recommended
by the ofcial agricultural services for cultivation in southern Brazil,
were also investigated. This study was carried out in accordance
with the current Brazilian legislation on biodiversity usage (Genetic
Heritage Management Council e Provisional Act 2.186-16, August
23, 2001) and is part of an agreement involving small farmer
communities of Santa Catarina State and the Federal University of
Santa Catarina.
2.2. Experimental design
A rst set of experiments used samples (500 g e dry weight) of
grains (hereafter named F0) obtained as previously described
(Section 2.1) aiming at to evaluate the physicochemical and thermal
properties of the maize our and starch fractions. In a second set of
experiments, eld trials were performed at the Experimental Field
of the Plant Science Center (Federal University of Santa Catarina,
Florianopolis, southern Brazil e 27
350 4800 S, 48
320 5700 W) by
cultivating the F0 maize seeds following typical agro-ecological
management adopted in southern Brazil for that cereal. In
a completely randomized design (DCC), maize seeds were sown in
August-2009 (1200 m2, 1 m  0.25 m e 4800 individuals). For each
maize varieties 600 individuals were sown in two consecutive lines.
The harvest period occurred in February-2010 and the grains (F1
progeny) were allowed to dry in the eld, followed by an ovendried treatment (65
C e 48 h) in laboratory.
2.3. Moisture content
The moisture content was determined following the method
described previously (AOAC, 1994).



Ca  Cv
U%
*100
a

(1)

where: (U) moisture content (%), (Ca) weight of crucible with

sample, (Cv) weight of crucible without sample, (a) weight of
sample (see Supplementary Table 1 e moisture content of the
maize genotypes).
2.4. Flour sample preparation
Seeds (250 g e dry weight) were selected from each maize
genotype and ground to pass a 0.5 mm sieve using a laboratory
cyclone mill (MB Braeski C.Q).
2.5. Starch isolation
Starch isolation was based on the methods described previously
(AOAC, 1994; Jane et al., 1999; Whistler, BeMiller, & Paschall, 1984)
with some modications. Briey, about 200 g of integral grain (8e
14% moisture content) were added to 100 ml of a 10% sodium
metabisulte solution. The mixture was maintained at room
temperature (25
C) for 48 h. After that, the steep water was
drained off, seeds were washed with distilled water and kept
soaked in water while removing the germ and the husk with the

616

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624

help of forceps. After removal, the grains were ground in a laboratory grinder with 600 ml of distilled water. The ground slurry was
ltered through a nylon cloth (100 mesh) and the residue was
washed with distilled water until to become free of starch. The
ltrate was passed successively over 200 and 325 mesh sieves and
the starcheprotein slurry was then allowed to stand for 30 min. The
supernatant was removed by suction and the settled starch layer
was resuspended in distilled water and centrifuged (3000 rpm,
20 min). The upper non-white layer was drained off and the white
layer was then collected and dried in an oven (45
C, 12 h) until
constant humidity (12%).
2.6. Physicochemical and functional properties
2.6.1. Swelling and solubility
Swelling power and solubility determinations were based on
Leach, McCowen, and Schoch (1959) with modications of the
methods previously described by Marcon, Avancini, & Amante,
2007; Adebooye & Singh, 2008; Aryee, Oduro, Ellis, & Afuakwa,
2006. Briey, a our sample (500 mg) was weighed into a 50 ml
graduated centrifuge tube. Distilled water was added to give a total
volume of 40 ml. The suspension was stirred sufciently and heated
at 95
C in a water bath for 30 min with constant stirring. The tubes
were cooled to room temperature and centrifuged (2500 rpm,
20 min). The supernatant was decanted carefully and 10 ml of the
residue was collected, transferred to a Petri dish (of known weight),
dried (60
C for 12 h), cooled and weighed for solubility index
determination. Solubility and swelling power were estimated with
the following equations:



Wr*Ww
SI%
*100
Ws*10

(2)

where: (SI-%) Solubility index; (Wr) weight of dry residue;

(Ww) weight of water; (Ws) weight of the sample.



Wg
SP%
*100
Wm

(3)

where: (SP-%) Swelling power; (Wg) weight of the gel;

(Wm) weight of the sample with moisture corrected.
2.6.2. Water binding capacity (WBC)
Water binding capacity (WBC) was determined by using the
described method (Yamazaki, 1953 with modications of Medcalf &
Gilles, 1965). An aqueous suspension was made by dissolving
500 mg of maize our in 10 ml of water. The suspension was
agitated for 30 min, followed by centrifugation (3000 rpm, 10 min).
The free water was decanted from the wet maize our, drained off
for 10 min and the wet sample weighed. WBC was estimated as
described previously (Yamazaki, 1953).



Dw  Ww
WBC% 1 
*100
Tw

(4)

where: (WBC) water biding capacity; (Dw) dry weight;

(Ww) wet weight; (Tw) total water added.
2.6.3. Fractionation of amylose
Amylose was separated from starch using a described method
(Mua & Jackson, 1995). Briey, using 10% aqueous n-butanol, starch
slurries (4% w/v) were prepared and heated at 45
C for 1 h with gentle
stirring. After dispersion, the slurry was centrifuged at 3000 rpm for
20 min. To precipitate amylose, 3 volumes of 100% n-butanol were
added. The mixture was swirled, incubated (2 h) at room temperature

and centrifuged in order to obtain the precipitate (amylose). Residues

obtained after the rst centrifugations were reslurried in methanol
and centrifuged to yield amylopectin. Both amylose and amylopectin
fractions were dried at 45
C in a forced-air oven.
2.6.4. Amylose
Amylose content of the isolated starches was determined
according the described method (Williams, Kuzina, & Hlynka, 1970).
Starch samples (20 mg) were taken and 10 ml of 0.5 N KOH were
added into it. The suspension was thoroughly mixed; the dispersed
sample was transferred to a 100 ml volumetric ask and diluted to
the mark with distilled water. An aliquot of the test starch solution
(10 ml) was pipetted into 50 ml volumetric ask and 5 ml of 0.1 N HCl
were added, followed by 0.5 ml of iodine reagent. The volume was
diluted to 50 ml and the absorbance measured at 625 nm (Gold
spectrum lab 53 UVeVis spectrophotometer, BEL photonics, Brazil).
The amylose content was determined from a standard curve
(y 0.09520x; r2 0.99) using amylose and amylopectin blends.
2.6.5. Scanning electron microscopy (SEM)
As previously described (Freitas, Paula, Feitosa, Rocha, &
Sierakowski, 2004), the defatting prior to preparation of our
samples was carried out by extraction under reux (ethyl ether, 6 h)
in a Soxleth apparatus. The defatted biomass was dried (100
C) and
ca. 10 mg of the our were mounted on double adhesive carbon
coated tape on an aluminum stub. The sample was coated with gold
using polaron E5001 SEM coating system. The coated sample was
then viewed under scanning electron microscopy (JEOL JSM6390LV model, JEOL Ltd., Tokyo, Japan) at 10 Kv and the micrographs were recorded with four replicates for each sample.
2.6.6. Fatty acid content
Lipid content was determined using the established method
(AOAC, 1994) as described (Cereda et al., 2002; Seabra, Zapata,
Nogueira, Dantas, & Almeida, 2002) and calculated according to
the following equation:



Wbl  Wb
LCmg=g
*100
W

(5)

where: (LC) lipid content; (Wbl) weight of balloon glass with

lipids; (Wb) weight of balloon glass, (W) weight of the sample.
2.7. Thermal properties
2.7.1. Differential scanning calorimetry (DSC)
Thermal characteristics of isolated starches were studied by
using DSC-50 Shimadzu, Japan, Model Shimadzu equipped with
a thermal analysis data station. Starch (13.0 mg dry weight) was
loaded into 40 ml capacity aluminum pan and distilled water was
added to achieve a starchewater suspension containing 30% water.
Samples were hermetically sealed and allowed to stand (1 h, room
temperature) before heating in a DSC. The DSC analyzer was calibrated using indium and an empty aluminum pan was used as
reference. Sample pan was heated from 20
C to 100
C, at 10
C/
min. Thermal transitions of starch samples were dened as T0
(onset), TP (peak of gelatinization) and Tc (conclusion). The gelatinization temperature range (R), was calculated as 2(TP  T0) as
described rstly (Freitas et al., 2004).
2.8. Pasting properties
2.8.1. Viscosity
Rheological behavior of maize ours was studied using a Rapid
Visco Analyser (RVA-4, Newport Scientic, Narabeen, Australia).

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624

617

Table 1
Physicochemical and functional properties of our of F0-maize landraces. Values expressed as mean  standard deviations of three independent experiments (n 3) are
presented for the protein, amylose and lipid contents, swelling power, solubility index, water binding capacity, and granule size. Distinct letters represent statistical differences
for the mean values at P < 0.05, by Tukey test.
Varieties
F0 progeny

Protein content
(g/100 g)

Amylose (mg/ml)

MPA1
PR
MG
LP
R8C
RJ
RXE
RX

9.07
10.59
8.54
11.59
7.04
9.28
10.55
11.02

7.74
5.30
8.69
8.52
15.85
14.32
5.58
9.60










0.16e
0.05g
0.18d
0.08d
0.01a
0.05b
0.06f
0.18c

Lipids (%)

Swelling
power (%)

3.29
3.74
5.53
3.38
4.46
3.10
3.01
3.02

9.88
8.74
9.57
8.07
10.52
8.44
9.82
8.70










1.87a
0.42b
1.73 ab
0.16d
1.28a
0.47c
0.59a
0.31b

Solubility index (%)

7.51
5.83
7.86
3.82
8.24
7.60
7.12
8.52










0.91b
2.66c
0.49ab
2.03d
0.50a
0.86b
0.22b
0.11a

Water binding
capacity (%)
18.17
18.89
18.83
18.36
17.43
17.56
18.13
18.19










1.86b
0.67a
0.81a
0.46ab
0.47c
0.78c
1.28b
0.23b

Granule sizea (mm)

11.80e18.80
8.20e11.80
8.40e12.00
10.00e15.20
7.20e16.40
7.20e10.20
9.40e16.60
4.80e13.60

Size of starch granules of the samples under SEM.

Flour samples (28 g e dry weight), suspended in deionized water

(8.0% m/v), were subjected to different timeetemperature proles,
e.g., heating from 48
C to 95
C with a stirring speed of 160 rpm
and incubation at 95
C/5 min. The viscosity was expressed in
centiPoise (cP, 0.1 kg m1 s1) and converted in mPa s
(1 cP 1 mPa s). The temperature of the onset in viscosity rise is the
pasting temperature. TPV is the temperature where the peak of
viscosity is reached. The breakdown is the difference between the
peak of viscosity and the minimum viscosity during pasting, and
setback is the difference between nal and minimum viscosities
during pasting.

(P < 0.05) as suitable for comparison of means among different

levels within a factor (Statistica v.6. and GraphPad Prism 5 statistical
packages). In all the experiments, the mean values are relative to a
minimum of three independent measurements (n 3). The ATRFTIR spectral data set and physicochemical variables were subjected
to multivariate analysis (PCAs and clustering) by using The R
statistical package (v.2.13.1). Previously to PCA analysis each spectrum within the 3000e600 cm1 region was standard normal
deviates corrected (Van Soest, Tournois, Wit, & Vliegen, 1995).
Analytical grade solvent and reagents were used in all the
experiments.

2.9. Fourier-transform infrared spectroscopy (FTIR)

3. Results and discussion
ATR-FTIR spectra of maize ours were recorded in a Bruker IFS-55
(Model Opus v. 5.0, Bruker Biospin, Germany) spectrometer with
a DTGS detector equipped with a golden gate single reection diamond attenuated total reectance (ATR) accessory (45
incidenceangle). A background spectrum of the clean crystal was acquired
and samples (100 mg) were spread and measured directly after
pressing them on the crystal. The spectra were recorded at the
absorbance mode from 4000 to 500 cm1 at the resolution of 4 cm1.
Five replicate spectra (128 co-added scans before Fourier transform)
were collected for each sample, in a total of 130 spectra. For processing the spectra were normalized, baseline-corrected in the region
of interest by drawing a straight line before resolution enhancement
(k factor of 1.7) was applied using Fourier self deconvolution (Cereda
et al., 2002; Krueger, Walker, Knutson, & Inglett, 1987; Rubens,
Snauwaert, Heremans, & Stute, 1999). The assumed line shape was

et al., 2006).
Lorentzian with a half width of 19 cm1 (Copkov
2.9.1. Statistical analysis
The data were examined statistically by using the one-way
analysis of variance (ANOVA), followed by the Tukey test

3.1. Physicochemical traits and functional properties

The amylose content ranged from 5.3 mg/ml (8.3%) to 15.9 mg/ml
(24.8%) for F0-our samples (Table 1) and from 7.9 mg/ml (12.4%) to
17.6 mg/ml (27.4%) for F1-samples (Table 2), as hybrid varieties
showed 7.6 mg/ml (11.9% e BR SC 154) and 19.5 mg/ml (30.4% e
Fortuna) of amylose. The landraces F0-R8C (24.8%) and F0-RJ
(22.4%) showed superior amounts (P < 0.05) of amylose for that
progeny in their grains. Regarding the F1-samples, PR landrace
showed the highest content of amylose (27.4%) and R8C (12.4%) the
lowest one, not differing from the MG-ML. Previous studies on
Argentinean maize varieties have been shown to contain amylose in
the range of 16.1e23.5% (Garcia et al., 1995) as Brazilian ML have
shown amylose amounts varying from 11.3 to 25.4% as previously
reported for our research group (Kuhnen et al., 2010). The amylose
content of starch granules varies with the botanical source and is
affected by climatic and soil conditions during grain development
(Sandhu, Singh, & Malhi, 2005). Amylose content of ML raw materials is recognized as an important trait because it affects the

Table 2
Physicochemical and functional properties of our of F1-maize landraces and hybrid varieties. Values expressed as mean  standard deviations of three independent
experiments (n 3) are presented for the protein, amylose and lipid contents, swelling power, solubility index, water binding capacity, and granule size. Distinct letters
represent statistical differences for the mean values at P < 0.05, by Tukey test.
Varieties

Protein content
(g/100 g)

Amylose (mg/ml)

Lipids (%)

Swelling power (%)

MPA1
PR
MG
LP
R8C
RJ
RXE
RX
BR SC 154
FORTUNA

6.70
6.99
6.05
6.96
5.60
6.18
7.05
7.18
9.52
6.03

12.97  0.10c
17.56  0.10a
8.05  0.01g
9.58  0.18f
7.91  0.05h
10.61  0.04e
14.11  0.05b
10.84  0.04d
7.64  0.03b
19.47  0.00a

5.03
4.30
4.74
5.43
4.15
4.58
5.03
4.96
6.65
4.97

9.08
8.89
9.55
12.25
11.46
13.14
10.87
9.85
10.88
10.87

Size of starch granules of the samples under SEM.

 0.47cd
 0.52d
 0.44c
 0.54a
 0.81b
 1.50a
 0.19b
 0.30c
 1.46a
 0.72a

Solubility index (%)

6.82
7.05
6.14
12.73
8.46
14.39
7.50
12.25
7.81
8.04

 0.16c
 0.48c
 2.13d
 1.09a
 1.09b
 3.11a
 1.14c
 1.12a
 0.67a
 0.12a

Water binding
capacity (%)
18.17
18.89
18.83
18.36
17.43
17.56
18.13
18.19
17.44
17.62

 1.86b
 0.67a
 0.81a
 0.46ab
 0.47c
 0.78c
 1.28b
 0.23b
 1.82a
 1.82a

Granule sizea (mm)

6.40e15.80
10.40e15.40
8.60e12.80
7.30e13.40
7.40e18.40
11.60e16.60
9.80e13.60
7.40e12.00
9.00e16.40
7.80e12.00

618

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624

functional properties of the starch such as swelling power and

solubility as further discussed.
Lipid contents varying from 3.01% (RXE) to 5.53% (MG) e F0
varieties e and from 4.15% (R8C) to 5.43% (LP) e F1 varieties e were
found (Tables 1 and 2). Interestingly, the highest amount of lipid
was detected for the BR SC 154 hybrid variety, i.e., 6.65%.
The moisture content (Supplementary Table 1) of the maize
genotypes varied from 8 to 13%, in accordance with the desired
value for analysis (below 14%) of the swelling power and solubility
index. The swelling power, solubility, water binding capacity
(WBC), and granule size are shown in Tables 1 and 2. Interestingly,
superior swelling powers were found for the F1-our samples RJ
(13.14  1.50), LP (12.25  0.54), and R8C (11.46  0.81), even higher
than those detected for the our samples of all the F0-ML. Similar
swelling power values were observed for the maize hybrid varieties, e.g., 10.88  1.46 e BR SC 154 and 10.87  0.72 e Fortuna.
The solubility index ranged from 3.82% (LP genotype) to 8.52%
(RX ML) for F0-maize our samples and from 6.14% (MG genotype)
to 14.39% (RJ ML) for the F1-ML progeny. The hybrid varieties
showed similar values of solubility index (P < 0.05).
Data of swelling power of maize starches (20.6%e24.5%) and
solubility (15.4%e21.7%) superior to those herein found have been
reported (Van Soest et al., 1995), mostly for starches extracted from
hybrid varieties as for ML information is scarce. The formation of
amyloseelipid complexes could be responsible for reducing the
solubility of starch (Kuhnen, 2007; Morrison, 1988), but we were
not able in nding a direct correlation between amyloseelipid
contents and their solubility indexes for the studied ML samples.
A large body of work has characterized amyloseelipid complexes in
terms of lipid type and, to lesser extent, starch type. Its known that
differences occur between the complexing ability of amylose and
amylopectin in different solutions, like as reported by Villwock,
Eliasson, Silverio, & BeMiller, 1999. On the other hand, the differences of swelling power and solubility among the samples could be
attributed to the presence of more brous materials in the whole
grain ours that possess more water holding capacity. In this study,
a direct relation between swelling power and moisture was
detected, as this behavior is attributed to the molecular

reorganization of starch caused by hydrothermal modication,

which provides an increased hydration of that macromolecule and
therefore enlarging the grain swelling (Olayinka, Adebowale, &
Olu-Owolabi, 2008). Besides, the swelling power of starches has
also been found to depend on the water binding capacity of starch
molecule by hydrogen bonding (Adebooye & Singh, 2008).
The discrepancies in solubility and swelling powers of starches
from different sources may also be due to differences in morphological structure of starch granules (Fig. 1). Higher swelling power and
lower solubility have been found in potato starches showing large and
irregular granules (Singh, Singh, Kaur, Sodhi, & Gill, 2003). Maize and
wheat granules may swell up to 30 times their original volume
without disintegration (Schoch, 1942). It has been suggested that
amylose plays a role in restricting initial swelling because this form of
swelling proceeds more rapidly after amylose has been exuded. The
increase in starch solubility, with concomitant suspension clarity is
seen mainly as the result of granule swelling. The extension of the
soluble leaching depends on the lipid content of the starch and the
ability of that polysaccharide to form amyloseelipid complexes.
Indeed, amyloseelipid complexes are insoluble in water and require
higher temperatures to dissociate (Kaur, Singh, & Sodhi, 2002). Taking
into account the lower values of solubility and swelling power herein
found, a strong interactions between lipids and starch components
seems to occur in the ML samples (Tables 1 and 2).
The discrepancies of swelling power and solubility of starches
among species and varieties are caused, for instance, by differences in
the amylose and lipid contents (Tables 1 and 2), as well as by the
granule structure (Fig. 1). The granules become increasingly susceptible to shear disintegration as they swell, releasing soluble material as
they disintegrate. The hot starch past is a mixture of swollen granules
and granule fragments, together with colloidal and molecularlydispersed starch granules. The mixture of swollen and fragmented
granules depends on the botanical source, water content, temperature and shearing during heating (Tester & Morrison, 1990). Finally,
water binding capacity (WBC) of the F0- and F1-maize ours in study
(Tables 1 and 2) presented similar values.
SEM analysis revealed a wide range of starch granule size for
both F0 (4.80e18.80 mm) and F1 progenies (6.40e18.40 mm e

Fig. 1. (AeD). Scanning electron microscopy (SEM) micrographic images of starch granules from grains of maize landraces. (A) Detail of starch grain of RJ-1. (B) Magnication of
previous gure showing spherical shape of the starch granule of individualized RJ-1. (C) Detail of starch grain of MG-1. (D) Magnication of previous gure showing the ellipsoid
shape of the individual bead array MG-1.

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624

Gelatinization temperatures

Onset To ( C)

Mid Tm ( C)

End Tc ( C)

LP-0
MG-0
MPA1-0
PR-0
R8C-0
RJ-0
RX-0
RXE-0

71.74
69.96
68.70
69.03
69.26
70.84
70.18
68.95

73.80
71.74
70.71
74.01
71.25
73.92
72.12
70.67

75.43
73.34
72.87
78.21
73.41
76.80
74.03
73.03

LP-1
MG-1
MPA1-1
PR-1
R8C-1
RJ-1
RX-1
RXE-1

74.51
75.88
67.23
67.15
75.85
67.93
66.10
66.72

76.88
77.06
69.88
70.36
76.86
71.42
69.73
68.95

78.76
78.40
72.64
73.58
77.92
74.48
73.62
73.49

BRSC154
FORTUN

74.09
67.62

76.42
69.99

78.73
72.35

100

600

Temp(C)
PR-0

MPA1-F0

R8C-0
RXE-0
MG-0

RX-0

RJ-0

LP-0

80

400
60

200
0

Temperature (C)

Varieties

800

Viscosity (mPa.s)

Table 3
Thermal properties of F0, F1 and hybrid maize starches under DSC analysis.
To onset temperature; Tm mid point; Tc nal temperature. Different letters
represent signicant differences for the mean values (Tukey test, P < 0.05) (see
Supplementary Fig. 1AeC for heat ow endotherms).

619

40
0

10
15
Time(min)

20

Fig. 2. Typical RVA pasting curves under Rapid Visco Analyser (RVA) from F0 maize
landraces (see Supplementary Fig. 2A and B for other amilographs of F1 and hybrid
varieties).

Wold, & Wold, 2001, 533 pp.; Schulz & Baranska, 2007). On the other
hand, such a trait seems to be inuenced by the region of cultivation
as herein detected. Indeed, a wide range of protein amount in the ML
grains was detected in our study from a given genotype, e.g., LP (F0,
11.59% x F1, 6.96%); RX (F0, 11.02% x F1, 7.18%).

Tables 1 and 2). Morphological details of the starch grains of the F1ML and hybrid varieties are shown in Fig. 1. Differences in swelling
power and solubility have also been attributed to changes in the
morphological granule size (Hormdok & Noomhorm, 2007).
Protein content of ML ours ranged from 7.04e11.59 g/100 g e F0
samples to 5.60e7.18 g/100 g e F1 samples as the hybrid varieties
presented 6.03 g/100 g e Fortuna and 9.52 g/100 g e BR SC 154
(Tables 1 and 2). The genotypes LP, RX (F0), and RX (F1) presented the
higher values of protein amounts. ML presented interesting values of
protein amounts as previously reported by our research group
comparatively to other studies. Protein contents varying from 5.7 to
8.9 g/100 g in maize hybrid varieties (Eriksson, Johansson, Kettaneh-

3.2. Thermal properties

Thermal properties of ML starches were studied through DSC
analysis. The transition temperatures (onset temperature e To, mid
point e Tm, and end set temperature e Tc) of maize starches are
shown in Table 3. To and Tc ranged from 66.1 to 75.9
C and 72.6 to
78.8
C, respectively. Similar results of To and Tc for maize starches
were also reported (Morrison, 1988). The lower To values were
observed for PR-F0 (69.0
C) and RX-F1 (66.1
C) as the ML LP-F0
(71.7
C) and R8C-F1 (75.9
C) showed higher values. Hybrid varieties ranged from 67.6
C to 74.1
C. Glass transition observed in our
study means the occurrence of amorphous constituents in the

Table 4
Pasting characteristics of maize landraces and hybrid varieties. Tpasting (
C) (temperature at viscosity rise), TPV (
C) (temperature at peak viscosity), peak viscosity (mPa s),
breakdown (mPa s) (peak viscosityeminimum viscosity), end viscosity (mPa s), setback (mPa s) (end viscosityeminimum viscosity), and minimum viscosity (mPa s) at 95
C of
F0, F1 and hybrid maize ours respectively.
Varieties

Pasting properties
Pasting
temperature (
C)

Temperature at
viscosity rise (
C)

Peak viscosity
(mPa s)

Final
viscosity
(mPa s)

Minimum
viscosity
(mPa s)

Breakdown
(mPa s)

Setback
(mPa s)

MPA1-0
PR-0
MG-0
LP-0
R8C-0
RJ-0
RXE-0
RX-0

69.85
87.95
91.60
69.90
95.05
75.15
74.70
79.95

48.95
49.05
48.75
48.95
48.90
49.20
49.05
49.05

193
188
343
200
212
179
188
212

125
228
291
125
772
446
176
145

12
37
112
49
94
103
36
46

181
151
231
151
18
76
152
166

113
191
179
76
578
343
140
99

MPA1-1
PR-1
MG-1
LP-1
R8C-1
RJ-1
RXE-1
RX-1

87.60
69.55
92.50
86.85
99.95
94.50
99.05
68.70

49.05
48.90
49.05
48.70
48.95
49.00
49.10
48.70

191
188
201
217
227
175
190
247

224
374
697
271
826
318
484
105

55
77
180
70
197
50
105
83

136
111
21
147
30
125
85
164

169
297
517
201
796
268
379
22

FORT
BR SC 154

91.60
75.95

49.05
48.95

170
177

251
159

40
16

130
161

211
143

620

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624

Fig. 3. (A) Principal component analysis (PCAs) scores scatter plot of ATR-FTIR spectra of Brazilian maize landraces and hybrid varieties ours. (B) Simple illustration of FTIR
spectrum of MPA-F0 ML (see Supplementary Fig. 3AeC for other FTIR images and Fig. 4 for hierarchical cluster dendogram).

starch and the presence of lipids that affect the starch gelatinization
(Dautant, Simancas, Sandoval, & Muller, 2007).
Our results of onset temperatures, gelatinization, and nal
temperatures can be explained based on the internal structure of
the molecules in the starch granule. Higher transition temperatures
result from higher degree of crystallinity, which provides structural
stability and makes the granules more resistant to gelatinization
(deWilligen, 1976). In fact, starches with higher content of

amylopectin, such as the ML samples in study, and longer branch

chain of that polysaccharide display the higher gelatinization
enthalpy, indicating that more energy is required to gelatinize the
crystallites of that starchy constituent. The discrepancies among
the starches from the ML in study are thought to be due to the
presence of crystalline regions of different strengths in the granules. DSC endotherms for the ML samples presented similar gelatinization temperatures (see Supplementary Fig. 1AeC).

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624

621

MG
RX1

LP1

2,4

1,8
Solubility

Breakdown

1,2
PeakVisc

RJ1

Lipids

FORT

Swelling

PC 2 (18.41%)

0,6
Wbinding

RX
-4

-3,2

-2,4

RXE1

-1,6

PR

-0,8

TPasting
1,6

0,8
Amylose

2,4

3,2

BR

Protein

-0,6 MPA1
GranuleS MG1

RXE MPA
LP

Setback

-1,2

R8C

GelatTemp

PR1
RJ
-1,8

R8C1
-2,4

-3
PC 1 (30.76%)

Fig. 4. Principal component analysis (PCAs) scores scatter plot of physicochemical variables of Brazilian maize landraces and hybrid varieties ours (see Supplementary Fig. 5AeC
for PCA loadings).

3.3. Pasting properties

The our samples in study showed pasting temperature ranging
from 68.7
C to 99.9
C. RX-F1 genotype showed the lower pasting
temperature (68.7
C) followed by MPA1-F0 (69.9
C). Contrarily,
the higher values of pasting temperature (w99
C) were detected
for R8C-F0/F1 and RXE-F1 ML. The setback was detected to vary
from 22 mPa s to 796 mPa s, with lower values for the F0-LP
(76 mPa s) and RX (99 mPa s). The F1-RX landrace showed even
lower setback, i.e., 22 mPa s, as the hybrid varieties presented
setback of 143 mPa s (BR SC 154) and 211 mPa s (Fortuna). Interestingly, the temperature at peak viscosity was similar for all the
maize ours samples. The higher viscosity was found for the MG-F0
(343 mPa s) and F1-RX (247 mPa s) genotypes as for the hybrid
varieties Fortuna presented superior pasting temperature (91.6
C),
but those genotypes were similar in their peak viscosities (Table 4).
The viscosity of cereal starch pastes is determined by lipids, mostly
phospholipids, creating complexes with amylose, slowing down or
even hindering granules swelling. Other effects are related to
decreased amylose solubility, retarded pasting, and limited gel
formation. Such complexes require higher temperatures to be subjected into dissociation (Morrison, 1988). Viscosity amilographs were
not discrepant for the ML (Fig. 2 and Supplementary Fig. 2AeB),
a nding thought to be related to the similar lipid content of those
genotypes (Tables 1 and 2). Finally, our ndings are in agreement with
previous studies on the viscosity of maize ours (Barichello, Yada,
Cofn, & Stanley, 1990; Shirai et al., 2007; Wu & Norton, 2001) so
that ML ours seem not to have a peculiar trait for that variable.
3.4. Attenuated total reectanceeFourier-transform infrared
spectroscopy (ATReFTIR)
Typical ATR-FTIR spectra of maize ours are presented in
Supplementary Fig. 3AeC for the F0, F1-progenies and hybrid

varieties, respectively. The visual analysis of the spectral prole

reveals the presence of many chemical constituents in the ngerprinting region (700e1500 cm1), where signals associated mostly
to the occurrence of proteins, lipids, amylose, and amylopectin
were clearly identied. Indeed, characteristic bands at 1635,
1670 cm1 were related to the presence of proteins as lipids/fatty
acids were detected at 2940, 2885, and 1750 cm1. Typical bands
associated to amylose (941 cm1) and amylopectin (1022 cm1)

were also detected (Berski et al., 2011; Copkov
et al., 2006;
Rubens et al., 1999).
3.5. Principal component and cluster analyses (PCAs)
In this study, PCA was used to objectively interpret and compare
the ATR-FTIR spectral data of the our samples in analysis and
evaluate the most important physicochemical variables able to
discriminate ML. The application of PCA for the spectral prole
(ngerprinting) allowed the large FTIR data set to be reduced to PC1
(50.90%) and PC2 (30.85%), which expressed 81% of the total variance of the spectral data set (Fig. 3). A clear separation of maize
landraces was observed, forming two groups of genotypes
according to their similarities along PC1 axis. Besides, the hybrid
varieties showed to be discrepants in their metabolic proles in
comparison to the ML in study, as determined by ATR-FTIR. Such
a nding is relevant taking into account the possibility of to explore
eventual new ML's chemical traits of signicance to food and
pharmaceutical industries, for instance. Interestingly, the PR ML
distinguished from all the genotypes analyzed, suggesting an even
more peculiar chemical composition and further studies in order to
elucidate its metabolic prole. mostly in respect to their lipid
contents (2796 cm1 and 2654 cm1) as indicated by the loading
values (data not shown). Further PCA analysis of the spectral
window associated to the lipid signals (3000e2800 cm1)
conrmed such ndings and revealed an additional grouping

622

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624

Distance
100

200

300

400

500

600

700

800

900

RJ
RXE1
R8C
MG1
R8C1
LP1
PR
MPA1
BR
PR1
RJ1
MG
RX1
RXE
FORT
LP
MPA
RX
Fig. 5. Similarity of maize landraces in respect to their physicochemical variables. Hierarchical cluster dendogram analysis UPGMA method with 71.74% of cophenetic correlation.
Varieties sufciently similar are represented in the same group and distinctions between groups. Vertical lines are used to connect branches at the similarity levels.

prole according to the content of those metabolites, corroborating

the chemical analysis (Tables 1 and 2). The MPA1-F0 ML was also
discriminated by PCA analysis from the other genotypes for the
carbohydrate region of the FTIR spectrum (1060 cm1e950 cm1).
The variable loadings correlated with PC1 were the bands at
1016 cm1 and 974 cm1, being these wave numbers related to the
distinction of amylose and amylopectin, respectively. On the other
hand, the ngerprint region to proteins (1650 cm1e1500 cm1)
and phenolic compounds (900 cm1e690 cm1) did not allow
a good discrimination for the genotypes in study. Similarities of
FTIR data with 71.74% of cophenetic correlation are represented in
cluster analysis (see Supplementary Fig. 4).
ATR/FTIR is a surface analytical method that can acquire information on the outer region of a sample as the infrared beam
penetrates into the rst few micrometers (w2 mm) of it. This
penetration depth is smaller than the average size of starch granules for the varieties studied which ranged from 4.80 mm to
18.80 mm (Tables 1 and 2). This implies that the IR spectra acquired
are representative of the external part of the our granules.
Therefore, it is believed that the differences in the infrared spectra
among these our samples are not related to the organization of the
starchs growth rings. The study of the surface and edge of starch
granules has been pointed out relevant to the understanding of the
relationship between the starch granule and its surrounding environment, for example (Freitas et al., 2004). Besides, the nature of
the granule surface with respect to crystallinity and absorbed nonstarch materials was suggested to delay enzyme action (Krueger
et al., 1987), an important trait to dene further industrial applications of our/starch biomasses. Thus, the variation among the
Z. mays genotypes in study might be interpreted in terms of the

level of ordered structure present on the edge of our granules,

suggesting the existence of discrepancies to their enzyme hydrolysis resistance.
When principal component analysis where performed for
physicochemical variables between ML, the rst three components
accounted for 30.71; 18.41 and 12% of the variation respectively
expressing 61.12% of total variance (Fig. 4). Swelling, pasting
temperature and setback were positively correlated with PC1 and
protein content, water binding content and breakdown with PC2
axis. The second component was correlated with solubility,
breakdown, peak viscosity, lipid content (PC2) and Gelatinization
temperature, setback and granule size (PC2). The third component was expressed with peak viscosity, water binding, setback
(PC3) and granule size, solubility and swelling (PC3) as represented by loadings (see Supplementary Fig. 5AeC). According to
variable plotting PCA allowed a clear separation between ML.
Hierarchical cluster analysis of physicochemical is showed in Fig. 5.
Similarities were dened on the basis of distance (Euclidean
distance) between two samples using unweighted arithmetic
average clustering method (UPGMA) as a suitable for this analysis.
Most similar varieties in their physicochemical variables are represented. A cophenetic correlation (similarity at which varieties
became members of the same cluster) was 71.75%.
4. Conclusions
The analytical approach herein described allowed detecting
interesting traits in ours and starches of ML cultivated in southern
Brazil. This way, industrial dedicated applications could be
attempted according to the genotype source of those peculiar

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624

biomasses. For instance, the genotypes RJ, RXE, and PR (F0 progeny)
can be used for purpose of industrial application such as elaboration of desserts (i.e., puddings) by their viscosities. Furthermore, RJ
and R8C (F0 progeny) are not desirable for the production of foods
that pass through storage with cycles of freezing and thawing for
their higher retrogradation. ATR-mid-infrared vibrational spectroscopy combined with chemometrics (PCAs) revealed discrepancies for the chemical prole of the genotypes related in any
extension to the ordered structure on the edge of our samples.
Such a nding is thought to be relevant for further analysis
considering the optimization of use of that raw material in food
industries, for instance. Besides, a clear separation of the ML was
found, revealing a peculiar chemical prole of that genotype, as
well as the effect of distinct environmental conditions of the
cultivation regions on the our samples chemical traits. Starches
and ours of F0-, F1-progeny ML presented desirable features so
that they can be used in agrifood industry to obtain certain products with peculiar characteristics. Such an approach corroborates to
add value and eventually new prospects for usage of maize landraces biomasses, encouraging the small farmer in southern Brazil to
preserve those valuable genetic resource materials.
Authors contributions
The rst author was the person who developed the work under
supervisoring of Prof. Marcelo Maraschin (last author e CNPq fellowshiper). Physico-chemical analyses were done under the guidance of the second author (Prof. Edna R. Amante). The viscosity
analyses were done by the third author (Prof. Ivo Demiate) and the
infrared analysis (ATR-FTIR), by the research group of Portugal
(fourth and fth author).
Acknowledgments
This research was carried out under CNPq-Brazil/MCTMozambique Postgraduate fellowship program. We are thankful to
Prof. Dr. Valdir Soldi and Marly Soldi (Laboratory of Polymers,
Department of Chemistry-UFSC), for their support in DSC analysis.
Special thanks to Electron Microscopy Central Laboratory (LCMEUFSC) and to Mr. Ricardo Brasil for his invaluable support in this
research.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in the
online version, at http://dx.doi.org/10.1016/j.foodhyd.2012.08.005.
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