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Physicochemical,thermal,andpastingpropertiesoffloursandstarchesofeightBrazilianmaizelandraces(ZeamaysL.)

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Food Hydrocolloids 30 (2013) 614e624

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Food Hydrocolloids

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

Physicochemical, thermal, and pasting properties of flours and starches of eightBrazilian 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, Brazilb Laboratory of Fruits and Vegetables, Departament of Food Science and Technology, Federal University of Santa Catarina, Florianopolis, BrazilcDepartment of Food Engineering, University of Ponta Grossa, Paraná, BrazildDepartment of Chemistry, University of Aveiro, Portugal

a r t i c l e i n f o

Article history:Received 5 December 2011Accepted 7 August 2012

Keywords:Maize landraces (Zea mays L.)ChemometricsStarchAmyloseViscositySolubilityATReFTIR

* Corresponding author. Tel.: þ5548 96128360; faxE-mail address: uaceleste@yahoo.com.br (V.G. Uar

0268-005X/$ e see front matter � 2012 Elsevier Ltd.http://dx.doi.org/10.1016/j.foodhyd.2012.08.005

a b s t r a c t

Both genetic and environmental factors create significant variation in the amount and quality of maizelandrace constituents. Details on the flours and starch characteristics have not been fully investigated.The physicochemical, pasting and thermal properties of 8 promising cultivars were assessed in this studyand those properties were correlated. Higher values of swelling and solubility (RJ e 13.14%; 14.39%), lipidcontent (MG e 5.53%), WBC (PR e 18.89%), and amylose content (PR e 27.43%) were found for thosegenotypes. 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 wasfound among the maize landraces (MG-F0, 343 mPa s and RJ-F1, 175 mPa s) as well as for the retro-gradation (R8C-F1, 796 mPa s and RX-F1, 22 mPa s). ATR-FTIR spectroscopy revealed amylose, amylo-pectin, lipids, and proteins as major flours constituents and their differences were discriminated by PCAanalysis.

� 2012 Elsevier Ltd. All rights reserved.

1. Introduction

Recent advances in biotechnology have accelerated the devel-opment and characterization of new crop genotypes and given risesome questions related to food security, conservation of plantgenetic resources, access and sustainable use of biological diversity,and environmental friendly agricultural productionmodels. Indeed,a significant part of agro-systems worldwide have moved fromsubsistence to intensive and market-oriented cultivation systemswhich commonly cultivate genetically improved varieties. Accord-ingly, local and creole genotypes (landraces e Zaid, Hughes,Porceddu, & Nicholas, 2001) have continuously been replaced incultivation systemsworldwide, leading to a reduction of the geneticdiversity of important crop species such as maize (Zea mays L. eLemos & Maraschin, 2008). This can result in an increased vulner-ability of that species to pest and diseases, restricting futureadaptive 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 agro-biodiversity (e.g., maize landraces e ML) are the replacement oflocal varieties by modern varieties, market integration and

: þ5548 37215400.rota).

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industrialization of agriculture including agro-processing andmarketing requirements (Drucker, Costa, & Magnusson, 2008).

In South hemisphere countries smallholder farmers have culti-vated hybrid Z. mays varieties to provide raw materials for agro-industry and, in parallel, maintain agro-ecological productionsystems of ML which have a current and meaningful local use inhuman and animal nutrition. Such genotypes have been selectedover generation for stability under low-external input conditionsand adverse environments and generally could not compete withnew high-yielding varieties bred for intensive production systems(FAO, 1998, p. 510). Besides, ML generally show distinct character-istics as to the standard established by the market, speciallyregarding their agronomic traits and continuous offering of rawmaterial (e.g., grains and flours), with difficulties to entering thecommercial circuit. This weak commerce appeal makes difficulttheir cultivation as some initiatives are needed to support smallfarmers interested in keeping them under crop. On the other hand,at present there is a paradox characterized by the food standardi-zation of one side, with over 75% of human food being based in lessthan a ten species, and on the other side the elevation of thedemand for more diversified, natural, healthful and organically(syn. biologically) foods such as ML flours (Vogt, 2005, 116 p.).

In this context, the creation of strategies to maintain landraceson cultivation is crucial and one possible approach for that involves

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624 615

the increase of smallholder farmer’s incomes. Thus, a suitableapproach seems to be the characterization of the landraces’ rawmaterials regarding their chemical traits that industrial applica-tions might be envisaged and developed. Such an approach isthought to be an add-value alternative to increase the economicsignificance of those genotypes, directly contributing for theimprovement of the family agriculture sector and preserving thatimportant germplasm. Indeed, some landraces have been used forthe development of new commercial varieties with desired char-acteristics that may result in innovations on food, chemical, phar-maceutical, and personal care industries.

Maize flours typically present a high starch and a wide range offood and non-food applications. Starches, because of their desirablephysical and chemical properties, are known to be suitable for use asthickeners, extenders, stabilizers, gelling agents, dietary calories,and texture modifiers in food formulations (Adebooye & Singh,2008). To enhance the suitability of different products, starch ismodified using chemical modifications techniques, e.g., hydrox-ypropylation, acetylation and cross-linking (Kaur, Singh, & Singh,2004). However, the use of chemically modified starches in foodproducts is either inhibited or discouraged in a number of countries.

Starches are found as water-insoluble granules of varying sizesand characteristic shapes depending on their botanical source. Theyare composed by a linear a-(1/4)-glucan, amylose (AM) and an a-(1/ 6)-branched a-(1/ 4)-glucan, amylopectin (AP). Such starchcomponents differ in their secondary and tertiary structures, givingrise to distinct arrangements of the crystalline and amorphousareas in the granules, which impart variability to the chemical andphysical properties of that polysaccharide (Mukerjea & Robyt,2010).

Amylose is used in the textile industry as a sizing and fishingagent, 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 industrypurposes because of the chemical residues left by fractionationmethods. To overcome this constraint, high-amylose maize geno-types (up to 70% amylose) have been developed. On the other hand,the branch chain length of amylopectin has been related to affectstarch crystallization (Garcia, Martino, & Zaritzky, 1995), gelatini-zation and pasting properties (Singh, Inouchi, & Nishinari, 2005).

Studies on the functional properties of the starch have beencarried out mostly using hybrid varieties of maize, as reports on thephysicochemical properties of flours and starches of maize land-races are scarce. Such genotypes are claimed to show high geneticvariability, a trait that might be correlated in any extension toa hypothesized chemical diversity, i.e., distinct types of starcheswith interesting possibilities for industrial applications. Besides, inseveral cases, the focus of investigation onmaize has been on starchgel viscoelasticity, while details on the flours (of whole grain ordecorticated grain) and starch characteristics have not been fullyinvestigated, especially for maize landraces. This study wasdesigned to investigate the physicochemical and thermal proper-ties of eight Brazilian ML, focusing on their flours and starchyfractions. The results are expected to be useful in providing guid-ance on the potential for industrial uses of those biomasses,improving the knowledge on theML biomasses and stimulating theproduction 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-

western region of Santa Catarina State, southern Brazil (Anchietacounty, 26�3101100 S, 53�2002600 W). Eight genotypes were selectedfor this study according to their importance for local populations asfollows: 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 byfarmers at Anchieta county based on Brazilian MPAeMovimento dePequenos Agricultores) and Língua de Papagaio (LP). For purpose ofcomparison as to the physicochemical characteristics in study, twomaize hybrid varieties, e.g., BR SC 154 and Fortuna, recommendedby the official agricultural services for cultivation in southern Brazil,were also investigated. This study was carried out in accordancewith the current Brazilian legislation on biodiversity usage (GeneticHeritage Management Council e Provisional Act 2.186-16, August23, 2001) and is part of an agreement involving small farmercommunities of Santa Catarina State and the Federal University ofSanta Catarina.

2.2. Experimental design

A first set of experiments used samples (500 g e dry weight) ofgrains (hereafter named F0) obtained as previously described(Section 2.1) aiming at to evaluate the physicochemical and thermalproperties of the maize flour and starch fractions. In a second set ofexperiments, field trials were performed at the Experimental Fieldof the Plant Science Center (Federal University of Santa Catarina,Florianopolis, southern Brazil e 27�3504800 S, 48�3205700 W) bycultivating the F0 maize seeds following typical agro-ecologicalmanagement adopted in southern Brazil for that cereal. Ina completely randomized design (DCC), maize seeds were sown inAugust-2009 (1200 m2, 1 m � 0.25 m e 4800 individuals). For eachmaize varieties 600 individuals were sown in two consecutive lines.The harvest period occurred in February-2010 and the grains (F1progeny) were allowed to dry in the field, followed by an oven-dried treatment (65 �C e 48 h) in laboratory.

2.3. Moisture content

The moisture content was determined following the methoddescribed previously (AOAC, 1994).

�Uð%Þ ¼ Ca� Cv

a*100

�(1)

where: (U) ¼ moisture content (%), (Ca) ¼ weight of crucible withsample, (Cv) ¼ weight of crucible without sample, (a) ¼ weight ofsample (see Supplementary Table 1 e moisture content of themaize genotypes).

2.4. Flour sample preparation

Seeds (250 g e dry weight) were selected from each maizegenotype and ground to pass a 0.5 mm sieve using a laboratorycyclone mill (MB Braeski C.Q).

2.5. Starch isolation

Starch isolationwas based on the methods described previously(AOAC, 1994; Jane et al., 1999; Whistler, BeMiller, & Paschall, 1984)with some modifications. Briefly, about 200 g of integral grain (8e14% moisture content) were added to 100 ml of a 10% sodiummetabisulfite solution. The mixture was maintained at roomtemperature (�25 �C) for 48 h. After that, the steep water wasdrained off, seeds were washed with distilled water and keptsoaked in water while removing the germ and the husk with the

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624616

help of forceps. After removal, the grains were ground in a labora-tory grinder with 600 ml of distilled water. The ground slurry wasfiltered through a nylon cloth (100 mesh) and the residue waswashed with distilled water until to become free of starch. Thefiltrate was passed successively over 200 and 325 mesh sieves andthe starcheprotein slurry was then allowed to stand for 30min. Thesupernatant was removed by suction and the settled starch layerwas resuspended in distilled water and centrifuged (3000 rpm,20 min). The upper non-white layer was drained off and the whitelayer was then collected and dried in an oven (45 �C, 12 h) untilconstant humidity (12%).

2.6. Physicochemical and functional properties

2.6.1. Swelling and solubilitySwelling power and solubility determinations were based on

Leach, McCowen, and Schoch (1959) with modifications of themethods previously described by Marcon, Avancini, & Amante,2007; Adebooye & Singh, 2008; Aryee, Oduro, Ellis, & Afuakwa,2006. Briefly, a flour sample (500 mg) was weighed into a 50 mlgraduated centrifuge tube. Distilled water was added to give a totalvolume of 40ml. The suspensionwas stirred sufficiently and heatedat 95 �C in a water bath for 30 minwith constant stirring. The tubeswere cooled to room temperature and centrifuged (2500 rpm,20 min). The supernatant was decanted carefully and 10 ml of theresidue was collected, transferred to a Petri dish (of knownweight),dried (60 �C for 12 h), cooled and weighed for solubility indexdetermination. Solubility and swelling power were estimated withthe following equations:

�SIð%Þ ¼ Wr*Ww

ðWs*10Þ*100�

(2)

where: (SI-%) ¼ Solubility index; (Wr) ¼ weight of dry residue;(Ww) ¼ weight of water; (Ws) ¼ weight of the sample.

�SPð%Þ ¼ Wg

Wm*100

�(3)

where: (SP-%) ¼ Swelling power; (Wg) ¼ weight of the gel;(Wm) ¼ weight of the sample with moisture corrected.

2.6.2. Water binding capacity (WBC)Water binding capacity (WBC) was determined by using the

describedmethod (Yamazaki, 1953 withmodifications of Medcalf &Gilles, 1965). An aqueous suspension was made by dissolving500 mg of maize flour in 10 ml of water. The suspension wasagitated for 30 min, followed by centrifugation (3000 rpm, 10 min).The free water was decanted from the wet maize flour, drained offfor 10 min and the wet sample weighed. WBC was estimated asdescribed previously (Yamazaki, 1953).

�WBCð%Þ ¼ 1� ðDw�WwÞ

ðTwÞ *100�

(4)

where: (WBC) ¼ water biding capacity; (Dw) ¼ dry weight;(Ww) ¼ wet weight; (Tw) ¼ total water added.

2.6.3. Fractionation of amyloseAmylose was separated from starch using a described method

(Mua & Jackson, 1995). Briefly, using 10% aqueous n-butanol, starchslurries (4%w/v)werepreparedandheatedat45 �C for1hwithgentlestirring. After dispersion, the slurry was centrifuged at 3000 rpm for20 min. To precipitate amylose, 3 volumes of 100% n-butanol wereadded. Themixturewas swirled, incubated (2h) at room temperature

and centrifuged in order to obtain the precipitate (amylose). Residuesobtained after the first centrifugations were reslurried in methanoland centrifuged to yield amylopectin. Both amylose and amylopectinfractions were dried at 45 �C in a forced-air oven.

2.6.4. AmyloseAmylose content of the isolated starches was determined

according the describedmethod (Williams, Kuzina, & Hlynka,1970).Starch samples (20 mg) were taken and 10 ml of 0.5 N KOH wereadded into it. The suspensionwas thoroughly mixed; the dispersedsample was transferred to a 100 ml volumetric flask and diluted tothe mark with distilled water. An aliquot of the test starch solution(10ml)waspipetted into 50mlvolumetricflask and5mlof 0.1NHClwere added, followed by 0.5 ml of iodine reagent. The volume wasdiluted to 50 ml and the absorbance measured at 625 nm (Goldspectrum lab 53 UVeVis spectrophotometer, BEL photonics, Brazil).The amylose content was determined from a standard curve(y ¼ 0.09520x; r2 ¼ 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 floursamples was carried out by extraction under reflux (ethyl ether, 6 h)in a Soxleth apparatus. The defatted biomass was dried (100 �C) andca. 10 mg of the flour were mounted on double adhesive carboncoated tape on an aluminum stub. The samplewas coatedwith goldusing polaron E5001 SEM coating system. The coated sample wasthen viewed under scanning electron microscopy (JEOL JSM-6390LV model, JEOL Ltd., Tokyo, Japan) at 10 Kv and the micro-graphs were recorded with four replicates for each sample.

2.6.6. Fatty acid contentLipid content was determined using the established method

(AOAC, 1994) as described (Cereda et al., 2002; Seabra, Zapata,Nogueira, Dantas, & Almeida, 2002) and calculated according tothe following equation:

�LCðmg=gÞ ¼ ðWbl�WbÞ

ðWÞ *100�

(5)

where: (LC) ¼ lipid content; (Wbl) ¼ weight of balloon glass withlipids; (Wb) ¼weight of balloon glass, (W) ¼weight 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 witha thermal analysis data station. Starch (13.0 mg dry weight) wasloaded into 40 ml capacity aluminum pan and distilled water wasadded to achieve a starchewater suspension containing 30% water.Samples were hermetically sealed and allowed to stand (1 h, roomtemperature) before heating in a DSC. The DSC analyzer was cali-brated using indium and an empty aluminum pan was used asreference. Sample pan was heated from 20 �C to 100 �C, at 10 �C/min. Thermal transitions of starch samples were defined as T0(onset), TP (peak of gelatinization) and Tc (conclusion). The gelati-nization temperature range (R), was calculated as 2(TP � T0) asdescribed firstly (Freitas et al., 2004).

2.8. Pasting properties

2.8.1. ViscosityRheological behavior of maize flours was studied using a Rapid

Visco Analyser (RVA-4, Newport Scientific, Narabeen, Australia).

Table 1Physicochemical and functional properties of flour of F0-maize landraces. Values expressed as mean � standard deviations of three independent experiments (n ¼ 3) arepresented for the protein, amylose and lipid contents, swelling power, solubility index, water binding capacity, and granule size. Distinct letters represent statistical differencesfor the mean values at P < 0.05, by Tukey test.

VarietiesF0 progeny

Protein content(g/100 g)

Amylose (mg/ml) Lipids (%) Swellingpower (%)

Solubility index (%) Water bindingcapacity (%)

Granule sizea (mm)

MPA1 9.07 7.74 � 0.16e 3.29 9.88 � 1.87a 7.51 � 0.91b 18.17 � 1.86b 11.80e18.80PR 10.59 5.30 � 0.05g 3.74 8.74 � 0.42b 5.83 � 2.66c 18.89 � 0.67a 8.20e11.80MG 8.54 8.69 � 0.18d 5.53 9.57 � 1.73 ab 7.86 � 0.49ab 18.83 � 0.81a 8.40e12.00LP 11.59 8.52 � 0.08d 3.38 8.07 � 0.16d 3.82 � 2.03d 18.36 � 0.46ab 10.00e15.20R8C 7.04 15.85 � 0.01a 4.46 10.52 � 1.28a 8.24 � 0.50a 17.43 � 0.47c 7.20e16.40RJ 9.28 14.32 � 0.05b 3.10 8.44 � 0.47c 7.60 � 0.86b 17.56 � 0.78c 7.20e10.20RXE 10.55 5.58 � 0.06f 3.01 9.82 � 0.59a 7.12 � 0.22b 18.13 � 1.28b 9.40e16.60RX 11.02 9.60 � 0.18c 3.02 8.70 � 0.31b 8.52 � 0.11a 18.19 � 0.23b 4.80e13.60

a Size of starch granules of the samples under SEM.

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624 617

Flour samples (28 g e dry weight), suspended in deionized water(8.0% m/v), were subjected to different timeetemperature profiles,e.g., heating from 48 �C to 95 �C with a stirring speed of 160 rpmand incubation at 95 �C/5 min. The viscosity was expressed incentiPoise (cP, 0.1 kg m�1 s�1) and converted in mPa s(1 cP¼ 1mPa s). The temperature of the onset in viscosity rise is thepasting temperature. TPV is the temperature where the peak ofviscosity is reached. The breakdown is the difference between thepeak of viscosity and the minimum viscosity during pasting, andsetback is the difference between final and minimum viscositiesduring pasting.

2.9. Fourier-transform infrared spectroscopy (FTIR)

ATR-FTIR spectra ofmaize flourswere recorded in a Bruker IFS-55(Model Opus v. 5.0, Bruker Biospin, Germany) spectrometer witha DTGS detector equipped with a golden gate single reflection dia-mond attenuated total reflectance (ATR) accessory (45� incidence-angle). A background spectrum of the clean crystal was acquiredand samples (100 mg) were spread and measured directly afterpressing them on the crystal. The spectra were recorded at theabsorbancemode from4000 to 500 cm�1 at the resolution of 4 cm�1.Five replicate spectra (128 co-added scans before Fourier transform)were collected for each sample, in a total of 130 spectra. For pro-cessing the spectrawerenormalized, baseline-corrected in the regionof interest by drawing a straight line before resolution enhancement(k factor of 1.7) was applied using Fourier self deconvolution (Ceredaet al., 2002; Krueger, Walker, Knutson, & Inglett, 1987; Rubens,Snauwaert, Heremans, & Stute, 1999). The assumed line shape wasLorentzian with a half width of 19 cm�1 (�Copíková et al., 2006).

2.9.1. Statistical analysisThe data were examined statistically by using the one-way

analysis of variance (ANOVA), followed by the Tukey test

Table 2Physicochemical and functional properties of flour of F1-maize landraces and hybridexperiments (n ¼ 3) are presented for the protein, amylose and lipid contents, swellinrepresent statistical differences for the mean values at P < 0.05, by Tukey test.

Varieties Protein content(g/100 g)

Amylose (mg/ml) Lipids (%) Swelling

MPA1 6.70 12.97 � 0.10c 5.03 9.08 � 0PR 6.99 17.56 � 0.10a 4.30 8.89 � 0MG 6.05 8.05 � 0.01g 4.74 9.55 � 0LP 6.96 9.58 � 0.18f 5.43 12.25 � 0R8C 5.60 7.91 � 0.05h 4.15 11.46 � 0RJ 6.18 10.61 � 0.04e 4.58 13.14 � 1RXE 7.05 14.11 � 0.05b 5.03 10.87 � 0RX 7.18 10.84 � 0.04d 4.96 9.85 � 0BR SC 154 9.52 7.64 � 0.03b 6.65 10.88 � 1FORTUNA 6.03 19.47 � 0.00a 4.97 10.87 � 0

a Size of starch granules of the samples under SEM.

(P < 0.05) as suitable for comparison of means among differentlevels within a factor (Statistica v.6. andGraphPad Prism 5 statisticalpackages). In all the experiments, the mean values are relative to aminimum of three independent measurements (n ¼ 3). The ATR-FTIR spectral data set and physicochemical variableswere subjectedto multivariate analysis (PCAs and clustering) by using The Rstatistical package (v.2.13.1). Previously to PCA analysis each spec-trum within the 3000e600 cm�1 region was standard normaldeviates corrected (Van Soest, Tournois, Wit, & Vliegen, 1995).Analytical grade solvent and reagents were used in all theexperiments.

3. Results and discussion

3.1. Physicochemical traits and functional properties

The amylose content ranged from5.3mg/ml (8.3%) to 15.9mg/ml(24.8%) for F0-flour samples (Table 1) and from 7.9mg/ml (12.4%) to17.6 mg/ml (27.4%) for F1-samples (Table 2), as hybrid varietiesshowed 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 thatprogeny in their grains. Regarding the F1-samples, PR landraceshowed the highest content of amylose (27.4%) and R8C (12.4%) thelowest one, not differing from the MG-ML. Previous studies onArgentineanmaize varieties have been shown to contain amylose inthe range of 16.1e23.5% (Garcia et al., 1995) as Brazilian ML haveshown amylose amounts varying from 11.3 to 25.4% as previouslyreported for our research group (Kuhnen et al., 2010). The amylosecontent of starch granules varies with the botanical source and isaffected by climatic and soil conditions during grain development(Sandhu, Singh, & Malhi, 2005). Amylose content of ML raw mate-rials is recognized as an important trait because it affects the

varieties. Values expressed as mean � standard deviations of three independentg power, solubility index, water binding capacity, and granule size. Distinct letters

power (%) Solubility index (%) Water bindingcapacity (%)

Granule sizea (mm)

.47cd 6.82 � 0.16c 18.17 � 1.86b 6.40e15.80

.52d 7.05 � 0.48c 18.89 � 0.67a 10.40e15.40

.44c 6.14 � 2.13d 18.83 � 0.81a 8.60e12.80

.54a 12.73 � 1.09a 18.36 � 0.46ab 7.30e13.40

.81b 8.46 � 1.09b 17.43 � 0.47c 7.40e18.40

.50a 14.39 � 3.11a 17.56 � 0.78c 11.60e16.60

.19b 7.50 � 1.14c 18.13 � 1.28b 9.80e13.60

.30c 12.25 � 1.12a 18.19 � 0.23b 7.40e12.00

.46a 7.81 � 0.67a 17.44 � 1.82a 9.00e16.40

.72a 8.04 � 0.12a 17.62 � 1.82a 7.80e12.00

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624618

functional properties of the starch such as swelling power andsolubility as further discussed.

Lipid contents varying from 3.01% (RXE) to 5.53% (MG) e F0varietiese and from 4.15% (R8C) to 5.43% (LP)e F1 varietiesewerefound (Tables 1 and 2). Interestingly, the highest amount of lipidwas detected for the BR SC 154 hybrid variety, i.e., 6.65%.

The moisture content (Supplementary Table 1) of the maizegenotypes varied from 8 to 13%, in accordance with the desiredvalue for analysis (below 14%) of the swelling power and solubilityindex. 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-flour samples RJ(13.14�1.50), LP (12.25� 0.54), and R8C (11.46� 0.81), even higherthan those detected for the flour samples of all the F0-ML. Similarswelling power values were observed for the maize hybrid varie-ties, 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 flour samples and from 6.14% (MG genotype)to 14.39% (RJ ML) for the F1-ML progeny. The hybrid varietiesshowed similar values of solubility index (P < 0.05).

Data of swelling power of maize starches (20.6%e24.5%) andsolubility (15.4%e21.7%) superior to those herein found have beenreported (Van Soest et al., 1995), mostly for starches extracted fromhybrid varieties as for ML information is scarce. The formation ofamyloseelipid complexes could be responsible for reducing thesolubility of starch (Kuhnen, 2007; Morrison, 1988), but we werenot able in finding a direct correlation between amyloseelipidcontents and their solubility indexes for the studied ML samples.A large body of work has characterized amyloseelipid complexes interms of lipid type and, to lesser extent, starch type. Its known thatdifferences occur between the complexing ability of amylose andamylopectin in different solutions, like as reported by Villwock,Eliasson, Silverio, & BeMiller, 1999. On the other hand, the differ-ences of swelling power and solubility among the samples could beattributed to the presence of more fibrous materials in the wholegrain flours that possess more water holding capacity. In this study,a direct relation between swelling power and moisture wasdetected, as this behavior is attributed to the molecular

Fig. 1. (AeD). Scanning electron microscopy (SEM) micrographic images of starch granulesprevious figure showing spherical shape of the starch granule of individualized RJ-1. (C) Deshape of the individual bead array MG-1.

reorganization of starch caused by hydrothermal modification,which provides an increased hydration of that macromolecule andtherefore enlarging the grain swelling (Olayinka, Adebowale, &Olu-Owolabi, 2008). Besides, the swelling power of starches hasalso been found to depend on the water binding capacity of starchmolecule by hydrogen bonding (Adebooye & Singh, 2008).

The discrepancies in solubility and swelling powers of starchesfrom different sources may also be due to differences in morpholog-ical structure of starch granules (Fig. 1). Higher swelling power andlower solubility havebeen found inpotato starches showing large andirregular granules (Singh, Singh, Kaur, Sodhi, & Gill, 2003). Maize andwheat granules may swell up to 30 times their original volumewithout disintegration (Schoch, 1942). It has been suggested thatamylose plays a role in restricting initial swelling because this formofswelling proceeds more rapidly after amylose has been exuded. Theincrease in starch solubility, with concomitant suspension clarity isseen mainly as the result of granule swelling. The extension of thesoluble leaching depends on the lipid content of the starch and theability of that polysaccharide to form amyloseelipid complexes.Indeed, amyloseelipid complexes are insoluble in water and requirehigher temperatures to dissociate (Kaur, Singh,& Sodhi, 2002). Takinginto account the lower values of solubility and swelling power hereinfound, a strong interactions between lipids and starch componentsseems to occur in the ML samples (Tables 1 and 2).

The discrepancies of swelling power and solubility of starchesamong species and varieties are caused, for instance, by differences inthe amylose and lipid contents (Tables 1 and 2), as well as by thegranule structure (Fig. 1). The granules become increasingly suscep-tible to sheardisintegrationas theyswell, releasing solublematerialasthey disintegrate. The hot starch past is amixture of swollen granulesand granule fragments, together with colloidal and molecularly-dispersed starch granules. The mixture of swollen and fragmentedgranules depends on the botanical source, water content, tempera-ture and shearing during heating (Tester & Morrison, 1990). Finally,water binding capacity (WBC) of the F0- and F1-maize flours in study(Tables 1 and 2) presented similar values.

SEM analysis revealed a wide range of starch granule size forboth F0 (4.80e18.80 mm) and F1 progenies (6.40e18.40 mm e

from grains of maize landraces. (A) Detail of starch grain of RJ-1. (B) Magnification oftail of starch grain of MG-1. (D) Magnification of previous figure showing the ellipsoid

Table 3Thermal properties of F0, F1 and hybrid maize starches under DSC analysis.To ¼ onset temperature; Tm ¼ mid point; Tc ¼ final temperature. Different lettersrepresent significant differences for the mean values (Tukey test, P < 0.05) (seeSupplementary Fig. 1AeC for heat flow endotherms).

Varieties Gelatinization temperatures

Onset To (�C) Mid Tm (�C) End Tc (�C)

LP-0 71.74 73.80 75.43MG-0 69.96 71.74 73.34MPA1-0 68.70 70.71 72.87PR-0 69.03 74.01 78.21R8C-0 69.26 71.25 73.41RJ-0 70.84 73.92 76.80RX-0 70.18 72.12 74.03RXE-0 68.95 70.67 73.03

LP-1 74.51 76.88 78.76MG-1 75.88 77.06 78.40MPA1-1 67.23 69.88 72.64PR-1 67.15 70.36 73.58R8C-1 75.85 76.86 77.92RJ-1 67.93 71.42 74.48RX-1 66.10 69.73 73.62RXE-1 66.72 68.95 73.49

BRSC154 74.09 76.42 78.73FORTUN 67.62 69.99 72.35

0 5 10 15 20

0

200

400

600

800

40

60

80

100

MPA1-F0Temp(ºC)PR-0

R8C-0

RJ-0

RX-0RXE-0 LP-0MG-0

Time(min)

Visco

sity (m

Pa.s)

Tem

peratu

re (ºC

)

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

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624 619

Tables 1 and 2). Morphological details of the starch grains of the F1-ML and hybrid varieties are shown in Fig. 1. Differences in swellingpower and solubility have also been attributed to changes in themorphological granule size (Hormdok & Noomhorm, 2007).

Protein content ofML flours ranged from7.04e11.59 g/100 ge F0samples to 5.60e7.18 g/100 g e F1 samples as the hybrid varietiespresented 6.03 g/100 g e Fortuna and 9.52 g/100 g e BR SC 154(Tables1 and2). Thegenotypes LP, RX (F0), andRX (F1)presented thehigher valuesof protein amounts.MLpresented interestingvalues ofprotein amounts as previously reported by our research groupcomparatively to other studies. Protein contents varying from 5.7 to8.9 g/100 g inmaize hybrid varieties (Eriksson, Johansson, Kettaneh-

Table 4Pasting characteristics of maize landraces and hybrid varieties. Tpasting (�C) (temperaturbreakdown (mPa s) (peak viscosityeminimumviscosity), end viscosity (mPa s), setback (mF0, F1 and hybrid maize flours respectively.

Varieties Pasting properties

Pastingtemperature (�C)

Temperature atviscosity rise (�C)

Peak viscos(mPa s)

MPA1-0 69.85 48.95 193PR-0 87.95 49.05 188MG-0 91.60 48.75 343LP-0 69.90 48.95 200R8C-0 95.05 48.90 212RJ-0 75.15 49.20 179RXE-0 74.70 49.05 188RX-0 79.95 49.05 212

MPA1-1 87.60 49.05 191PR-1 69.55 48.90 188MG-1 92.50 49.05 201LP-1 86.85 48.70 217R8C-1 99.95 48.95 227RJ-1 94.50 49.00 175RXE-1 99.05 49.10 190RX-1 68.70 48.70 247

FORT 91.60 49.05 170BR SC 154 75.95 48.95 177

Wold, &Wold, 2001, 533 pp.; Schulz & Baranska, 2007). On the otherhand, such a trait seems to be influenced by the region of cultivationas herein detected. Indeed, awide rangeof protein amount in theMLgrains 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%).

3.2. Thermal properties

Thermal properties of ML starches were studied through DSCanalysis. The transition temperatures (onset temperature e To, midpoint e Tm, and end set temperature e Tc) of maize starches areshown in Table 3. To and Tc ranged from 66.1 to 75.9 �C and 72.6 to78.8 �C, respectively. Similar results of To and Tc for maize starcheswere also reported (Morrison, 1988). The lower To values wereobserved 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 vari-eties ranged from 67.6 �C to 74.1 �C. Glass transition observed in ourstudy means the occurrence of amorphous constituents in the

e at viscosity rise), TPV (�C) (temperature at peak viscosity), peak viscosity (mPa s),Pa s) (end viscosityeminimumviscosity), and minimumviscosity (mPa s) at 95 �C of

ity Finalviscosity(mPa s)

Minimumviscosity(mPa s)

Breakdown(mPa s)

Setback(mPa s)

125 12 181 113228 37 151 191291 112 231 179125 49 151 76772 94 18 578446 103 76 343176 36 152 140145 46 166 99

224 55 136 169374 77 111 297697 180 21 517271 70 147 201826 197 30 796318 50 125 268484 105 85 379105 83 164 22

251 40 130 211159 16 161 143

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

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624620

starch and the presence of lipids that affect the starch gelatinization(Dautant, Simancas, Sandoval, & Muller, 2007).

Our results of onset temperatures, gelatinization, and finaltemperatures can be explained based on the internal structure ofthe molecules in the starch granule. Higher transition temperaturesresult from higher degree of crystallinity, which provides structuralstability 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 branchchain of that polysaccharide display the higher gelatinizationenthalpy, indicating that more energy is required to gelatinize thecrystallites of that starchy constituent. The discrepancies amongthe starches from the ML in study are thought to be due to thepresence of crystalline regions of different strengths in the gran-ules. DSC endotherms for the ML samples presented similar gela-tinization temperatures (see Supplementary Fig. 1AeC).

Protein

Amylose

Lipids

Swelling

Solubility

Wbinding

GranuleS

TPasting

PeakViscBreakdown

Setback

GelatTemp

MPA

PR

MG

LP R8C

RJ

RXE

RX

MPA1

PR1

MG1

LP1

R8C1

RJ1

RXE1

RX1

BR

FORT

-4 -3,2 -2,4 -1,6 -0,8 0,8 1,6 2,4 3,2

PC 1 (30.76%)

-3

-2,4

-1,8

-1,2

-0,6

0,6

1,2

1,8

2,4

PC2

(18.

41%

)

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

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624 621

3.3. Pasting properties

The flour samples in study showed pasting temperature rangingfrom 68.7 �C to 99.9 �C. RX-F1 genotype showed the lower pastingtemperature (68.7 �C) followed by MPA1-F0 (69.9 �C). Contrarily,the higher values of pasting temperature (w99 �C) were detectedfor R8C-F0/F1 and RXE-F1 ML. The setback was detected to varyfrom 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 evenlower setback, i.e., 22 mPa s, as the hybrid varieties presentedsetback of 143 mPa s (BR SC 154) and 211 mPa s (Fortuna). Inter-estingly, the temperature at peak viscosity was similar for all themaize flours samples. The higher viscosity was found for theMG-F0(343 mPa s) and F1-RX (247 mPa s) genotypes as for the hybridvarieties Fortuna presented superior pasting temperature (91.6 �C),but those genotypes were similar in their peak viscosities (Table 4).

Theviscosityof cereal starchpastes isdeterminedby lipids,mostlyphospholipids, creating complexes with amylose, slowing down oreven hindering granules swelling. Other effects are related todecreased amylose solubility, retarded pasting, and limited gelformation. Such complexes require higher temperatures to be sub-jected into dissociation (Morrison,1988). Viscosity amilographswerenot discrepant for the ML (Fig. 2 and Supplementary Fig. 2AeB),a finding thought to be related to the similar lipid content of thosegenotypes (Tables 1 and2). Finally, ourfindings are inagreementwithprevious studies on the viscosity of maize flours (Barichello, Yada,Coffin, & Stanley, 1990; Shirai et al., 2007; Wu & Norton, 2001) sothat ML flours seem not to have a peculiar trait for that variable.

3.4. Attenuated total reflectanceeFourier-transform infraredspectroscopy (ATReFTIR)

Typical ATR-FTIR spectra of maize flours are presented inSupplementary Fig. 3AeC for the F0, F1-progenies and hybrid

varieties, respectively. The visual analysis of the spectral profilereveals the presence of many chemical constituents in the finger-printing region (700e1500 cm�1), where signals associated mostlyto the occurrence of proteins, lipids, amylose, and amylopectinwere clearly identified. Indeed, characteristic bands at 1635,1670 cm�1 were related to the presence of proteins as lipids/fattyacids were detected at 2940, 2885, and 1750 cm�1. Typical bandsassociated to amylose (941 cm�1) and amylopectin (1022 cm�1)were also detected (Berski et al., 2011; �Copíková et al., 2006;Rubens et al., 1999).

3.5. Principal component and cluster analyses (PCAs)

In this study, PCAwas used to objectively interpret and comparethe ATR-FTIR spectral data of the flour samples in analysis andevaluate the most important physicochemical variables able todiscriminate ML. The application of PCA for the spectral profile(fingerprinting) allowed the large FTIR data set to be reduced to PC1(50.90%) and PC2 (30.85%), which expressed 81% of the total vari-ance of the spectral data set (Fig. 3). A clear separation of maizelandraces was observed, forming two groups of genotypesaccording to their similarities along PC1 axis. Besides, the hybridvarieties showed to be discrepants in their metabolic profiles incomparison to the ML in study, as determined by ATR-FTIR. Sucha finding is relevant taking into account the possibility of to exploreeventual new ML's chemical traits of significance to food andpharmaceutical industries, for instance. Interestingly, the PR MLdistinguished from all the genotypes analyzed, suggesting an evenmore peculiar chemical composition and further studies in order toelucidate its metabolic profile. mostly in respect to their lipidcontents (2796 cm�1 and 2654 cm�1) as indicated by the loadingvalues (data not shown). Further PCA analysis of the spectralwindow associated to the lipid signals (3000e2800 cm�1)confirmed such findings and revealed an additional grouping

900

800

700

600

500

400

300

200

100

Distance

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 sufficiently similar are represented in the same group and distinctions between groups. Vertical lines are used to connect branches at the similarity levels.

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624622

profile according to the content of those metabolites, corroboratingthe chemical analysis (Tables 1 and 2). The MPA1-F0 ML was alsodiscriminated by PCA analysis from the other genotypes for thecarbohydrate region of the FTIR spectrum (1060 cm�1e950 cm�1).The variable loadings correlated with PC1 were the bands at1016 cm�1 and 974 cm�1, being these wave numbers related to thedistinction of amylose and amylopectin, respectively. On the otherhand, the fingerprint region to proteins (1650 cm�1e1500 cm�1)and phenolic compounds (900 cm�1e690 cm�1) did not allowa good discrimination for the genotypes in study. Similarities ofFTIR data with 71.74% of cophenetic correlation are represented incluster analysis (see Supplementary Fig. 4).

ATR/FTIR is a surface analytical method that can acquire infor-mation on the outer region of a sample as the infrared beampenetrates into the first few micrometers (w2 mm) of it. Thispenetration depth is smaller than the average size of starch gran-ules for the varieties studied which ranged from 4.80 mm to18.80 mm (Tables 1 and 2). This implies that the IR spectra acquiredare representative of the external part of the flour granules.Therefore, it is believed that the differences in the infrared spectraamong these flour samples are not related to the organization of thestarch’s growth rings. The study of the surface and edge of starchgranules has been pointed out relevant to the understanding of therelationship between the starch granule and its surrounding envi-ronment, for example (Freitas et al., 2004). Besides, the nature ofthe granule surface with respect to crystallinity and absorbed non-starch materials was suggested to delay enzyme action (Kruegeret al., 1987), an important trait to define further industrial appli-cations of flour/starch biomasses. Thus, the variation among theZ. mays genotypes in study might be interpreted in terms of the

level of ordered structure present on the edge of flour granules,suggesting the existence of discrepancies to their enzyme hydro-lysis resistance.

When principal component analysis where performed forphysicochemical variables between ML, the first three componentsaccounted for 30.71; 18.41 and 12% of the variation respectivelyexpressing 61.12% of total variance (Fig. 4). Swelling, pastingtemperature and setback were positively correlated with PC1þ andprotein content, water binding content and breakdown with PC2�axis. The second component was correlated with solubility,breakdown, peak viscosity, lipid content (PC2þ) and Gelatinizationtemperature, setback and granule size (PC2�). The third compo-nent was expressed with peak viscosity, water binding, setback(PC3þ) and granule size, solubility and swelling (PC3�) as repre-sented by loadings (see Supplementary Fig. 5AeC). According tovariable plotting PCA allowed a clear separation between ML.Hierarchical cluster analysis of physicochemical is showed in Fig. 5.Similarities were defined on the basis of distance (Euclideandistance) between two samples using unweighted arithmeticaverage clustering method (UPGMA) as a suitable for this analysis.Most similar varieties in their physicochemical variables are rep-resented. A cophenetic correlation (similarity at which varietiesbecame members of the same cluster) was 71.75%.

4. Conclusions

The analytical approach herein described allowed detectinginteresting traits in flours and starches of ML cultivated in southernBrazil. This way, industrial dedicated applications could beattempted according to the genotype source of those peculiar

V.G. Uarrota et al. / Food Hydrocolloids 30 (2013) 614e624 623

biomasses. For instance, the genotypes RJ, RXE, and PR (F0 progeny)can be used for purpose of industrial application such as elabora-tion of desserts (i.e., puddings) by their viscosities. Furthermore, RJand R8C (F0 progeny) are not desirable for the production of foodsthat pass through storage with cycles of freezing and thawing fortheir higher retrogradation. ATR-mid-infrared vibrational spec-troscopy combined with chemometrics (PCAs) revealed discrep-ancies for the chemical profile of the genotypes related in anyextension to the ordered structure on the edge of flour samples.Such a finding is thought to be relevant for further analysisconsidering the optimization of use of that raw material in foodindustries, for instance. Besides, a clear separation of the ML wasfound, revealing a peculiar chemical profile of that genotype, aswell as the effect of distinct environmental conditions of thecultivation regions on the flour samples’ chemical traits. Starchesand flours of F0-, F1-progeny ML presented desirable features sothat they can be used in agrifood industry to obtain certain prod-ucts with peculiar characteristics. Such an approach corroborates toadd value and eventually new prospects for usage of maize land-races biomasses, encouraging the small farmer in southern Brazil topreserve those valuable genetic resource materials.

Author’s contributions

The first author was the person who developed the work undersupervisoring of Prof. Marcelo Maraschin (last author e CNPq fel-lowshiper). Physico-chemical analyses were done under the guid-ance of the second author (Prof. Edna R. Amante). The viscosityanalyses were done by the third author (Prof. Ivo Demiate) and theinfrared analysis (ATR-FTIR), by the research group of Portugal(fourth and fifth author).

Acknowledgments

This research was carried out under CNPq-Brazil/MCT-Mozambique Postgraduate fellowship program. We are thankful toProf. 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 (LCME-UFSC) and to Mr. Ricardo Brasil for his invaluable support in thisresearch.

Appendix A. Supplementary data

Supplementarydataassociatedwith thisarticle canbe found, in theonline version, at http://dx.doi.org/10.1016/j.foodhyd.2012.08.005.

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