Accepted Manuscript

Characterization of corn (Zea mays L.) bran as a new food ingredient for snack bars

Milena Figueiredo de Sousa, Rafaiane Macedo Guimarães, Marcos de Oliveira

Araújo, Keila Rezende Barcelos, Nárgella Silva Carneiro, Daniele Silva Lima,
Daiane Costa Dos Santos, Karla de Aleluia Batista, Kátia Flávia Fernandes, Mayra
Conceição Peixoto Martins Lima, Mariana Buranelo Egea

PII: S0023-6438(18)31049-1
DOI: https://doi.org/10.1016/j.lwt.2018.11.088
Reference: YFSTL 7652

To appear in: LWT - Food Science and Technology

Received Date: 27 March 2018

Revised Date: 26 November 2018
Accepted Date: 28 November 2018

Please cite this article as: Figueiredo de Sousa, M., Guimarães, R.M., de Oliveira Araújo, M., Barcelos,
K.R., Carneiro, Ná.Silva., Lima, D.S., Dos Santos, D.C., de Aleluia Batista, K., Fernandes, Ká.Flá.,
Peixoto Martins Lima, Mayra.Conceiçã., Egea, M.B., Characterization of corn (Zea mays L.) bran
as a new food ingredient for snack bars, LWT - Food Science and Technology (2018), doi: https://
doi.org/10.1016/j.lwt.2018.11.088.

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to
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1 CHARACTERIZATION OF CORN (Zea mays L.) BRAN AS A NEW FOOD

2 INGREDIENT FOR SNACK BARS

4 Milena Figueiredo de Sousa12, Rafaiane Macedo Guimarães1, Marcos de Oliveira Araújo1,

5 Keila Rezende Barcelos1, Nárgella Silva Carneiro1, Daniele Silva Lima1, Daiane Costa Dos

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6 Santos1, Karla de Aleluia Batista34, Kátia Flávia Fernandes4, Mayra Conceição Peixoto

Martins Lima1, Mariana Buranelo Egea1*

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9 1 Programa de Pós-Graduação em Tecnologia de Alimentos, Instituto Federal de Educação,

10 Ciência e Tecnologia Goiano – Campus Rio Verde, Rio Verde, GO, Brazil

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2 Faculdade Morgana Potrich (FAMP) – Mineiros, GO, Brazil
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12 3 Instituto Federal de Educação, Ciência e Tecnologia de Goiás – Campus Goiânia Oeste,
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13 Goiânia, GO, Brazil

14 4 Laboratório de Química de Proteínas, Departamento de Bioquímica e Biologia Molecular,

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15 Instituto de Ciências Biológicas, Universidade Federal de Goiás, Goiânia, CP131, CEP

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16 74001-970, GO, Brazil

17 * Corresponding author: Goiano Federal Institute of Education, Science and Technology,

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18 Campus Rio Verde, Rod. Sul Goiana, Km 01, CP 66, CEP 75901-970, Rio Verde, GO, Brazil

Phone no.: +55 64 3620 5600; e-mail address: mariana.egea@ifgoiano.edu.br

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20 Abstract

21 This study aimed to demonstrate the potential of a corn byproduct, namely corn bran (CB) as

22 a new source of nutrients in the diet by evaluating its application to snack bars (SB). CB was

23 separated into five granulometric measurements (F1: 0.125; F2: 0.250; F3: 0.5; F4: 1.0; and

24 F5: 2.0 mm) that had different profiles: F5 showed the highest dietary fiber content (42%), F4

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25 had the highest moisture content (11%), F3 exhibited the highest protein content (13%) and

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26 F2 had the highest ash content (3.47%). Concerning absorption properties in water, milk and

27 oil, F3, F4 and F5 exhibited the highest values, whereas F1 and F2 had the highest solubility

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28 in water, besides foaming and emulsifying activity. F3 fraction (1.0 mm) was chosen for its

29 physico-chemical characteristics for the development of SB. In SB formulation, CB was

30
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substituted in the following proportions in the mass of dry ingredients: 0% (SB0, control),
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31 10% (SB10), 25% (SB25), 40% (SB40) and 55% (SB55). Results of chemical analyses
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32 demonstrated that SB formulations with the highest proportions of CB had the highest levels

33 of dietary fiber. Furthermore, it should be highlighted that SB made with CB have expressive
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34 nutritional value that are important to health maintenance.

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35 Keywords: product development, dietary fiber, functional foods

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36 1 Introduction
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38 Cereals play an important role in human nutrition, whether for direct consumption or

39 as a raw material for flour production. Therefore, corn (Zea mays L.) is one of the major

40 cereals cultivated in the world, followed by wheat and rice (De Vasconcelos et al., 2013). The

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41 2016/17 corn harvest, which yielded 1075.22 million tons, was 9.5% higher than the 2015/16

42 one and represented an increase that was strongly influenced by the boost in supply in South

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43 America, especially in Brazil (USDA, 2018).

Expansion of the global corn market has led to the production of a large amount of

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45 corn bran (CB), a byproduct of the milling industry. Even though CB has traditionally been

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46 used in animal feed, it can be applied to human food, since it is rich in dietary fiber (Rose,
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47 Inglett, & Liu, 2010). Consequently, all ingredients which are rich in dietary fiber provide

48 functional properties, such as water retention capacity and ability to absorb fecal mutagens
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49 (due to their high dietary fiber content), and protect the human body against their harmful

50 effects (Zeng & Lazarova, 2014).

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51 Snack bars (SB), also known as cereal bars, have been widely consumed all over the
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52 world because they are low-calorie snack options which are easy to carry (Suhem, Matan,
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53 Matan, Danworaphong, & Aewsiri, 2015). SB are commercially and nutritionally interesting

54 not only because they mitigate immediate hunger, but also because they favorably influence
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55 people’s nutritional status (Trier & Johnston, 2012). Most consumers are people who care for
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56 their diet and health and eat SB as sources of some nutrients, such as fiber, protein, minerals

57 and vitamins (Boustani & Mitchell, 1990).

58 Food industries have been using strategies to supplement products with ingredients

59 that are vehicles for functional compounds, such as plant-based products and byproducts

60 (Helkar, Sahoo, & Patil, 2016). SB may be important vehicles for transporting these

61 ingredients and providing bioactive compounds to the human diet (Sun-Waterhouse, Teoh,
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62 Massarotto, Wibisono, & Wadhwa, 2010). Thus, this study aimed to evaluate CB as a novel

63 food ingredient for SB production.

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65 2 Material and Methods

66 2.1 Plant material and snack bar (SB) formulation

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67 CB was obtained by the dry milling process of an agro-industry. The byproduct was

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68 separated into five granulometric measurements (0.125; 0.250; 0.5; 1.0; and 2.0 mm) and

69 fractions were named F1, F2, F3, F4 and F5, respectively. Due to its physical-chemical

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70 characteristics, F3 (1.0 mm) was chosen for SB development. The SB formulation (Table 1)

71 was previously tested for complete agglomeration of solid ingredients. CB substitution in SB

72
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formulation was performed in the following proportions in the mass of dry ingredients: 0%
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73 (SB0, control), 10% (SB10), 25% (SB25), 40% (SB40) and 55% (SB55). Ingredients were
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74 heated at 60 ± 3 ºC, mixed, laminated, cut and stored in polyethylene plastic bags at room

75 temperature until analysis (Figure 1).

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77 2.2 Chemical and biochemical composition

78 Proximal composition (moisture, protein, lipids, ashes, carbohydrates by difference) of

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79 CB fractions and SB was performed in agreement with AOAC (2007) methods. The total

dietary fiber in CB was determined by a K-TDFR-Megazyme enzyme kit, while dietary fiber
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81 (DF), insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) in SB were evaluated by

82 the enzymatic-gravimetric method. Caloric value was calculated by the Atwater conversion

83 factor (Merril & Watt, 1973).

84 For mineral quantification, samples were dried in an oven at 105ºC and digested in

85 HNO3 and HClO4. Regarding minerals in CB, calcium (Ca), magnesium (Mg), copper (Cu),

86 iron (Fe), manganese (Mn) and zinc (Zn) were determined by dual beam atomic absorption
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87 spectroscopy GBC-XPLORAA-2 (Laeside, Australia). Potassium (K) was determined by

88 flame photometer B-462 (Micronal), phosphorus (P) and sulfur (S) were determined by

89 molecular absorption spectrophotometry (Silva, 2009). Sodium quantification in SB was

90 performed by a Shimadzu Atomic Absorption Spectrometer AA-6300 (Kyoto, Japan) with

91 background correction by a Deuterium Lamp.

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92 Fatty acid composition in SB was determined by CP 3800 Gas Chromatography

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93 (Varian) with a DB-23 capillary column (60 m, 0.25 D, 0.25 µm thick). Gas fluxes were 1.3

94 mL/min for H2 entrainment gas, 30 mL/min for auxiliary gas N2 and 30 and 300 mL/min for

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95 H2 flame gas and synthetic air, respectively. Sample split ratio was 1:100. The temperature of

96 the column was programmed at 150 ºC/16 min, raised to 180 ºC at a rate of 2 ºC/min and kept

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at this temperature for 20 min. Injector and detector temperatures were 250 and 280 °C,
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98 respectively.
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99 To evaluate the inhibitory activity of trypsin, analyses proposed by Arnon (1970) were

100 performed. Briefly, sample of corn bran extract (1% w⁄v) were incubated with 0.1 mL trypsin
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101 0.5 mg/mL solution and 0.1 mol/L phosphate buffer (pH 7.6) at 37 ºC/10 min and 1 mL of a
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102 casein solution 1% (w⁄v) was added to the solution. The reaction was interrupted after 10

103 minutes by adding of 1.5 mL of TCA 10% (v ⁄ v) solution. Trypsin inhibitory unit (TIU) was
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104 defined as the difference between the units observed in the maximum activity and the activity

of the samples containing the inhibitors. Activity of α-amylase inhibitor was determined in
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106 agreement with Deshpande, Sathe, Salunkhe, & Cornforth (1982), using starch as the

107 substrate for the enzyme. Phytic acid content was determined by the method described by

108 Latta & Eskin (1980) with modifications for resin DOWEX-AGX-4, as proposed by Ellis &

109 Morris (1986). A standard curve of phytic acid (Sigma, P8810) was done and results were

110 expressed as percentage (g/100g) of the sample. Hemagglutination assays, with the use of

111 rabbit erythrocytes, were performed by the method described by Moreira & Perrone (1977),
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112 with modifications. CB extract (1% w/v) was diluted in two-fold dilution series against a 0.15

113 mol/L NaCl solution. One milliliter of a 2% erythrocyte suspension was added to an equal

114 volume of the sample and the mixture was incubated at 37 ºC/30 min. Then, it was kept at

115 room temperature (25 ºC/30 min). Tubes were centrifuged at 3000 rpm/1min and the last tube

116 to show visible agglutination was considered the point of equivalence.

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117

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118 2.3 Physical and technological characterization

119 A Color Flex EZ spectrophotometer (Hunterlab, Virginia, USA) was employed to

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120 determine the color of CB fractions and SB, in agreement with the CIE (Commission

121 Internationale de I'Eclairage) system (standard illuminant D65/10°). The microstructural

122
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analysis of CB fractions with increments of 100x, 300x, 2,000x and 8,000x was conducted by
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123 an electronic scanning electron microscope (acceleration voltage 5 kV, JSM-6610/Jeol®)
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124 equipped with EDS (Thermo Scientific NSS Spectral Imaging) which used degreased

125 samples, dried at 40 °C.

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126 Water solubility (WS), water absorption index (WAI), milk absorption index (MAI)
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127 and oil absorption capacity (OAC) were determined by the methods described by Okezie &

128 Bello (1988). Least gelation capacity (LGC) and foaming properties were determined in
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129 agreement with Coffmann & Garcia (1977). In the analysis of foaming properties, a CB

suspension (3 g flour in 150 mL distilled water) was prepared and whipped at 10.000 rpm for
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131 5 min by a HMD200 mixer (Hamilton Beach, Virginia, USA). Immediately afterward, the

132 mixture was transferred to a 250 mL graduated cylinder. Volumes were recorded before and

133 after the whipping process. The percentage ratio of the increased volume, by comparison with

134 the initial volume, was calculated before whipping as the index of foam capacity (FC). The

135 sample beaker was kept at room temperature and foam stability (FS) was evaluated by

136 checking volume decrease at 30, 60, 90 and 120 min. Emulsifying capacity (EC) was
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137 determined in agreement with Yasumatsu et al. (1972). The calculation was done by dividing

138 the height of the emulsified layer by the total height.

139 Specific volumes of CB formulations were determined by the displacement of millet

140 seed, as proposed by Goula & Adamopoulos (2008), using millet seed. SB texture was

141 performed by a TA-XT Plus texturometer (Texture Analyzer, Surrey, England), as described

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142 by Alam et al. (2017), who analyzed maximum shear strength (tangential force) with blade set

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143 with guillotine HDP/BSG, using velocity of 0.5 mm/s, distance of 15 mm and the limit force

144 of 0.05 N.

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145

146 2.4 Microbiological analyses and sensory evaluation

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Microbiological analyses (Bacillus cereus per gram, Coliforms at 45 ºC and
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148 Salmonella sp. by 25 grams) were performed by standard protocols (Doores, Salfinger, &
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149 Tortorello, 2013).

150 Sensory analyses were performed by 80 panelists (84% female and 16% male)
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151 between 17 and 49 years old. They were students, professors and employees at the Morgana
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152 Potrich School (FAMP) located in Mineiros, GO, Brazil. The sensory evaluation was

153 previously approved by the Research Ethics Committee at the Faculdades Integradas de Santa
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154 Fé do Sul (CAAE no. 58184016.4.0000.5428). SB were served in individual booths and the

preference sorting test was used. Participants were asked to order the samples from the lowest
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156 to the highest preference.

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158 2.5 Statistical analysis

159 Data were subjected to analysis of variance (ANOVA) to detect significant differences

160 among treatments whereas means were compared by the Tukey’s test using Statistica 6.0

161 software (StatSoft Inc., USA). Differences were considered significant when p<0.05. All
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162 experiments were performed in triplicate. Sensory results were performed by the Friedman

163 test and by Newell & MacFarlane (1987).

164

165 3 Results and Discussion

166 3.1 Chemical and biochemical composition

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167 CB fractions with the highest yield in the separation came from sieves with 0.5 (F3);

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168 0.25 (F2) and 1 mm (F4) meshes. There were differences among them (totaling 96.45%).

169 There was no retention in the aperture sieve size of 0.063 mm; therefore, this fraction was not

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170 considered in the other evaluations. Singh, Liu, & Vaughn (2012) obtained 91% particle

171 retention in 0.251 and 0.354 mm meshes and stated that particle size is an important criterion

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in the case of raw ingredients, since it may affect the quality and taste of food products.
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173 Granulometric fractions had different profiles of proximal composition (Table 2). The
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174 highest protein content was assigned to F3, whereas the highest ash content was provided by

175 the F2 fraction. CB is the byproduct of corn milling for the production of brewers’ grits. The
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176 largest fractions (F4 and F5) exhibited corn hull (fiber), while small-sized particles (F1, F2
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177 and F3 fractions) were composed of particles resulting from corn grain breaking and polishing

178 (the internal part) (see the graphical abstract).

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179 Dietary fiber content of fractions varied between 8.60 and 42.11 g/100g. The larger the

mesh, the higher the dietary fiber content. It shows that CB can be a good source of dietary
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181 fiber in the human diet. Several studies have shown that dietary fiber intake plays a protective

182 role against obesity and other chronic diseases. In addition, CB-rich diets have led to

183 significant improvement towards pain severity, stool frequency, stool consistency, number of

184 additional gastrointestinal symptoms and total symptom score in patients with irritable bowel

185 syndrome (Cook et al., 1990).

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186 Agro-industrial byproducts that are rich in dietary fiber, such as CB, are excellent raw

187 materials for the food industry, since consumers prefer natural supplements to synthetic

188 ingredients (Bharat Helkar, Sahoo, & Patil, 2016; Rose et al., 2010). Supplementation with

189 ingredients which are rich in dietary fiber can result in health-promoting foods which have

190 low calories, low cholesterol and low fat, besides changes in textural properties (Elleuch et

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191 al., 2011; Zeng & Lazarova, 2014).

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192 Macro and micro mineral contents of CB fractions are shown in Table 3. The highest

193 iron contents were found in F1 and F2 fractions (4.71 and 4.47 mg/100g, respectively),

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194 significantly different from the other fractions. In relation to the phosphorus content, the

195 highest amounts were observed in fractions F1, F2 and F3 fractions, whereas the lowest ones

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were found in fractions F4 and F5. Phosphorus contents of F1, F2 and F3 fractions were
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197 compatible with the RDI for adults. The F2 fraction had the highest potassium content
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198 (870.00 mg/100 g), significantly different from the other fractions. The highest magnesium

199 content was found in the F2 fraction (306.67 g/100 g), whereas the lowest one was exhibited
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200 by the F5 fraction (93.33 g/100 g). Regarding the sulfur content, there was no significant
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201 difference among F2, F3, F4 and F5 fractions. Concerning manganese and copper, F5 had the

202 highest contents. F1, F2 and F3 fractions did not differ from each other; they had higher zinc
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203 content than the one reported for quinoa (Chenopodium quinoa W.) (2.48 g/100 g) (Li & Zhu,

2017). Therefore, 100 g CB contributes with 50% copper and zinc contents to the RDI in the
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205 diet of adults. Variations in the mineral content found in corn may be due to genetic and/or

206 environmental factors (Sokrab, Mohamed Ahmed, & Babiker, 2012).

207 Table 4 shows antinutritional factors of CB fractions. The content of trypsin inhibitors

208 ranged from 1.88 to 3.08 IU/mg, close to results reported by Batista, Prudêncio, & Fernandes

209 (2010) for beans flour. The phytic acid content was observed to increase as the sieve meshes

210 decreased (from F5 to F1); besides, it was lower than the one reported for peach palm (6.66
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211 and 6.34 mg/g) (Bolanho, Danesi, & Beléia, 2013). Phytic acid is considered an

212 antinutritional factor since it forms complexes with proteins and minerals, mainly cations of

213 potassium, magnesium, calcium, iron and zinc, and reduces their bioavailability. However,

214 this property confers antioxidant function to phytic acid because of its iron chelating ability

215 and prevents the mineral from catalyzing the formation of hydroxyl groups, thus, resulting in

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216 benefits to human health (Jenab & Thompson, 2002).

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217 The fact that no haemagglutinating activity was detected in CB fractions represents a

218 positive result, since agglutination of blood cells reduces efficiency of the oxygen transport

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219 system. This result shows that, in the samples, there is neither lectin nor proteins of this class

220 that can induce hemagglutination (Peumans & Van Damme, 1995). Lecithin is a compound

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which is considered toxic because it may cause ulceration and necrosis of intestinal villi
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222 (Oliveira, Vidal, & Sgarbieri, 1989).
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224 3.2 Physical and technological characteristics

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225 Table 5 shows the chromatic analysis and technological properties of CB. In the
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226 chromatic analysis, values of a* positive were less expressive than values of b* positive,

227 indicating that the CB predominant color is yellow (28.92-31.93). Values of the hue angle
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228 decreased as fraction size increased. Values were between 80 and 87, indicating yellow color.

229 Luminosity ranged from 71.11 to 83.50. The smaller the CB fraction, the higher the L* value.
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230 These values of luminosity are within the range found in the literature for dry corn flour
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231 (78.1-87.4) (Lozano-Alejo, Carrillo, Pixley, & Palacios-Rojas, 2007). Results of chroma

232 values were higher (31.90-32.16) in F2 and F4 fractions, thus showing high intensity of

233 yellow in these fractions.

234 Table 5 shows absorption properties in water, milk and oil. The F5 fraction had the

235 highest values, by comparison with the other fractions. This behavior may be due to the

236 contact surface and the high dietary fiber content found in this fraction (Table 2). Conversely,
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237 the smallest particles (<0.5 mm) were more soluble in water, and consequently, both FC and

238 EC were higher. According to Wang, Rocha, Nascimento, & Ascheri (2006), the highest WAI

239 values are more desirable in bakery products because they allow the addition of more water to

240 the dough and contribute to improve handling characteristics which, in this case, would be

241 associated with the smallest fractions of corn bran (F1, F2 and F3 fractions).

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242 High results of foam and emulsion properties may be related not only to particle size,

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243 but also to high protein value (8.26 - 12.39%), as shown by Siddiq, Ravi, Harte, & Dolan,

244 (2010). Furthermore, Zheng, Yang, Tang, Li, & Ahmad (2008) reported that foam capacity is

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245 determined by the amount and nature of proteins used for making foaming easier and

246 reducing surface tension at the air/ water interface.

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Hydration properties of dietary fiber are related to particle size, shape and ionic
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248 strength, pH, temperature, chemical structure of polysaccharides and porosity. Likewise, fiber
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249 has oil absorption capacity which is mainly related to the surface properties of bran particles

250 and to the overall charge density and hydrophilic nature of constituents (Elleuch et al., 2011).
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251 There was gelatinization in all CB fractions; minimum concentrations were 4% for the
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252 F5 fraction and 2% for the other fractions. They were enough to compose the gelled matrix.

253 The gelatinization phenomenon results from the formation of a carbohydrate three-
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254 dimensional network, partially denatured protein molecules and lipids (Adebowale & Lawal,

2003). Consequently, F1, F2 and F3 fractions can be applied to foods, such as mousse, bread,
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256 meringue and ice cream, which require foaming, emulsifying and gelatinizing properties. The

257 F4 fraction was chosen for SB development since it combines both high yield and fiber

258 content (30.6 g/100 g).

259 Images of the scanning electron micrograph analysis (SEM) (Fig.2) showed large

260 structural differences among fractions with the largest particles (F5) and the other fractions.

261 The F5 fraction was found to have a flat and rectilinear surface in relation to others that have
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262 flattened and rounded forms. De la Hera, Talegón, Caballero, & Gómez (2013) also found

263 differences in the microstructure of fractions of different types and particle sizes of corn flour.

264 They observed compact leaf fragments in fractions >180µm and loose fragments with few

265 intergranular spaces in the smallest fractions.

266 In agreement with what was reported by Uarrota et al. (2013) for corn starch, in this

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267 study, few granules and small starch sizes were observed in the F5 fraction, whereas larger

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268 granules were found in other CB fractions. Moreover, the smaller the CB particle size, the

269 smoother the surface of the starch granules (neither irregularities nor superficial porosity were

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270 found in spherical structures and small size particles).

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272 3.3 Snack bar (SB) development

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273 SB proximal composition is shown in Table 6. Moisture contents ranged between 8.42
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274 and 9.55 g/100 g. The higher the added CB, the higher the ash content in SB (1.00 - 1.22

275 g/100 g). As a result, SB55 was significantly different from the other formulations. The ash
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276 content of all SB formulation was higher than the one reported by Zamora-Gasga, Bello-
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277 Pérez, Ortíz-Basurto, Tovar, & Sáyago-Ayerdi (2014) for granola bars with agave syrup (0.95

278 g/100 g). It is important because increase in ash content implies increase in mineral content.
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279 Amounts of total dietary fiber with prevalence of insoluble dietary fiber found in CB

were significantly different among formulations under investigation, since they were
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281 relatively proportional to the increase in the CB proportion in the formulations (as expected

282 with increase in CB) (Table 6). Values found by this study in the formulations were higher

283 than those reported by Sun-Waterhouse et al. (2010) for cereal bars with apple fiber (0.34

284 g/100 g). Furthermore, the US Food and Drug Administration allows foods to be labeled as ‘a

285 good source of fiber’ or ‘high fiber’ if they contain from 2.5 g to 5.0 g dietary fiber per
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286 serving, respectively (FAO, 2008). Considering 40 g SB per serving, SB35, SB40 and SB55

287 can be considered and labeled ‘good sources of fiber’.

288 These results confirm the potential this byproduct has to increase fiber contents in

289 food products. Physiological benefits of fiber to the body include reduction of cardiovascular

290 diseases, obesity, some types of cancer and intestinal constipation. Fiber also favors the

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291 immune system, acts in weight reduction, increases postprandial energy expenditure and leads

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292 to excretion of bile acids (Brownlee, Chater, Pearson, & Wilcox, 2017; Zeng & Lazarova,

293 2014).

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294 The highest lipid content was found in SB55 (2.64 g/100 g), significantly different

295 from the other treatments, without significant differences among them. Saturated fat contents

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in the formulations under development were higher in F25 and F55 (without significant
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297 difference) (1.09 and 1.14 g/100 g, respectively), which differed from the other formulations
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298 under study. No amount of trans fat content was found in the snack bars developed by this

299 study. With regards to the sodium content, the highest value was found in SB0 (110.29
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300 mg/100 g), which only differed significantly from SB40 (79.04 mg/100 g). They were lower
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301 than values reported by Possinger (2014) for peanut butter and chocolate cereal bars and

302 peanut and granola bars (600 mg/100 g and 300 mg/100 g sodium, respectively). SB10, SB25
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303 and SB40 were less caloric (from 365 to 367 kcal/100 g), without significant difference

among them, than SB0 and SB55 (370.32 and 373.09 kcal/100 g, respectively). A 40% CB
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305 substitution in cereal bars results in almost 30% reduction in energy value, 90% saturated fat

306 and 60% sodium, by comparison with commercial snack bars consumed in the United

307 Kingdom (Minister of Agriculture and Agri-Food, 2013). These three factors (energy value,

308 saturated fat and sodium content) should be evaluated together (John et al., 2016), since they

309 have been controlled by regulatory agencies to avoid public health problems (Mayne &
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310 Spungen, 2017). In addition, foods containing few calories, besides little saturated fat and

311 sodium, have attracted consumers’ attention.

312 Physical parameters found for the five formulations under investigation are shown in

313 Table 7. In general, SB were rectangular, heterogeneous, yellowish, bright and crispy. Both

314 formulations SB40 and SB55 had the highest brightness (L*), differing from SB0, SB10 and

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315 SB25. There were no significant differences among formulations with respect to parameter

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316 a*, while an increase in parameter b* was observed when the amount of CB increased. As a

317 result, SB were yellow, a characteristic of CB (Table 5).

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318 The highest hardness was found in SB55 (126.26 N) whereas SB10 showed the lowest

319 hardness (76.78 N). Chen & Opara (2013) stated that texture is an attribute for quality control

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and product acceptance, especially when it comes to snack bars. Formulations with CB
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321 showed the highest hardness (between 76.78 and 126.26 N) by comparison with fruit bars
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322 elaborated by Parn, Bhat, Yeoh, & Al-Hassan (2015) (21.47 N). It may happen due to the

323 amount of carbohydrates in cereal products that can affect physical properties, mainly texture
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324 (Heenan et al., 2012).

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325 The Bacillus cereus count was <1.0x101 CFU/g in all formulations under evaluation.

326 Salmonella spp. was not found in CB formulations. Concerning thermotolerant coliforms
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327 (0.31 NMP/g), all formulations were within the parameters, thus, microbiologically safe.

According to Hausner, Hartvig, Reinbach, Wendin, & Bredie (2012), preference for
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328
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329 whole foods may be related to food choices which are usually guided by preferences that the

330 individual establishes early in life. Food acceptance is determined by perception and food

331 taste and texture. These characteristics may be modified by cognitive effects, appetite, food

332 situation and culture. In the group of sensory evaluators, 82% of the judges claimed to like

333 foods and products with whole grains, 36.25% rarely consume these products, 21.25%

334 consume them up to 3 times a week and 15% eat them 3 times per month.
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335 The preference sorting-test was used for verifying which SB formulation under

336 development would be the favorite one. Calculated F was 12.56, higher than tabulated F

337 (9.53), i. e., there were significant differences among formulations. In this case, the Newel

338 table was used for comparing formulations and verifying which samples differed from each

339 other (n=80, t=5). When differences between the frequency sum of the formulations evaluated

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340 with the table value were compared, significant difference was only observed between SB0

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341 and SB55 (minimum significant difference=55). Thus, substitution of dry ingredients at

342 concentrations up to 40% CB did not interfere with the preference for snack bars. Aigster,

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343 Duncan, Conforti, & Barbeau (2011) observed that granola bars with starch addition were

344 accepted by the judges, i. e., 68% demonstrated to like them (slightly or extremely). This

345
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positive result resembles the one found by this study and shows that snack bars are important
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346 vehicles for potentially functional ingredients.
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347

348 4 Conclusion
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349 CB fractions were shown to have potential for food application. All fractions had high
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350 fiber content, a fact that reinforces their advantages to food industry. SB formulations

351 developed by this study with CB in ratios up to 55% showed increase in nutritional value
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352 when there was increase in CB content. Formulations with up to 40% CB were well-accepted

by the judges, by comparison with the formulation without any CB. Thus, results show that
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353
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354 corn bran, a byproduct of the dry milling industry, can be used for developing food products

355 intended for human consumption.

356

357 Acknowledgments

358 The authors are grateful to FAPEG, CAPES, CNPq and IF Goiano for their financial

359 support.
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360

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519
520 Figure captions

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522 Figure 1. Snack bar development flow chart

523

524 Figure 2. Scanning electron spectrographs at 2000x (upper) and 8000x (bottom) of the five

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525 CB fractions
526

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Table 1. Ingredients (g) used in the production of snack bars with corn bran

Ingredients SB0 SB10 SB25 SB40 SB55

(0%) (10%) (25%) (40%) (55%)

Rice flakes 13.72 12.35 10.29 8.23 6.17

Whole oat flakes 18.75 16.87 14.05 11.24 8.33

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Thin oat flakes 19.00 17.09 14.24 11.39 8.54

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Corn bran 0.00 5.14 12.86 20.58 28.30

Raisins 29.30 29.30 29.30 29.30 29.30

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Corn glucose 28.46 28.46 28.46 28.46 28.46

Crystal sugar 26.03 26.03 26.03 26.03 26.03

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Table 2. Chemical composition of corn bran (CB)

Analyzes F1 F2 F3 F4 F5

Moisture (g/100g) 8.19±0.15d 8.48±0.04d 9.38±0.03c 11.66±0.19a 10.59±0.06b

Proteins (g/100g) 10.33±0.31c 11.62±0.31b 12.39±0.00a 8.08±0.31d 8.26±0.31d

Lipids (g/100g) 12.81±0.14b 13.85±0.10a 12.74±0.14b 8.91±0.10c 7.69±0.36d

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Ash (g/100g) 3.11±0.05b 3.47±0.02a 3.14±0.04b 1.78±0.04c 1.43±0.18d

56.97±1.51a 51.90±0.56b 46.35±0.21c 39,50±0.61d 29.92±0.77e

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Carbohydrates

TDF (g/100g) 8.60±1.18e 10.67±0.22d 16.00±0.35c 30.07±0.85b 42.11±0.77a

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Energy (kcal/100g) 418.91±0.99a 421.45±0.45a 413.60±0.65b 390.82±1.12c 390.40±2.30c

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Table 3. Mineral composition of corn bran and Recommended Daily Intake (RDI) for adults

Minerals RDI F1 F2 F3 F4 F5

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(mg/100g) (mg/d)

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Phosphorous 700(2) 707±5b 820±17a 710±10b 353±30c 260±10d

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Potassium 4700(3) 760±17b 870±36a 793±11b 487±11c 377±25d

30.0±0.0a 33.3±5.8a 30.0±0.0a 33.3±5.8a 26.7±15.3a

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Calcium 1000.0(1)

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Magnesium 260(1) 257±5b 307±5a 266±5b 130±10c 93±15d

Sulfur - 143±5b 163±15ab 147±5ab 167±5a 163±5ab

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Iron 14.00(1) 4.71±0.12a 4.47±0.08a 3.65±0.11b 2.48±0.04c 2.72±0.38c

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Manganese 2.30(2)R 0.76±0.02c 0.82±0.02bc 0.84±0.02b 0.80±0.01bc 0.96±0.04a

Copper 0.90(2) 0.45±0.01b

TE 0.48±0.01ab 0.43±0.00b 0.40±0.02b 0.59±0.10a
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Zinc 7.00(1)S 3.96±0.82a 3.85±0.04a 3.78±0.03a 2.59±0.07b 2.33±0.27b
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Triplicate average. Means followed by the same letter in the line do not differ from each other, according to the Tukey’s test at 5% significance.
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(1): FAO / WHO, 2001. (2) IOM (Institute of medicine), 2001. (3): IOM (Institute of medicine), 2005. R: 10% Bioavailability. S: Moderate

bioavailability - calculated on the basis of mixed diets containing animal protein.

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Table 4. Antinutritional factors from corn bran fractions

F1 F2 F3 F4 F5

Trypsin

inhibitors 2.31±0.05bc 2.07±0.13bc 2.65±0.41ab 1.88±0.30c 3.08±0.09a

(IU/mg)

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Trypsin
15.22±0.30b 13.62±0.85b 17.48±2.70a 12.39±1.96 20.28±0.59

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inhibition unit
c c b c a

(TIU, %)

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α-amilase

inhibitors 67.15±2.4a 63.13±3.5a 0.34±0.00b 1.21±0.00b 1.99±0.00b

(IU/mg)
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Phytic acid
47.84±0.00a 14.25±0.31b 6.85±0.09c 5.28±0.13d 0±0.00e
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(mg/g)

Hemagglutinatio
D

Abs Abs Abs Abs Abs

n activity
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IU: inhibition unit; Abs absent. Means followed by the same letter in the line do not

differ from each other, in agreement with the Tukey’s test at 5% significance.
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Table 5. Physical and technological characteristics of corn bran fractions

F1 F2 F3 F4 F5

L* 83.50±0.25a 80.97±0.12b 77.89±0.20c 74.43±0.41d 71.11±0.46e

a* 1.43±0.08e 2.54±0.06d 3.58±0.09c 4.28±0.37b 4.90±0.79a

b* 28.92±0.35c 31.80±0.36a 31.93±0.49a 31.87±0.59a 30.65±1.21b

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C* 28.95±0.35c 31.90±0.36ab 32.13±0.50a 32.16±0.63a 31.04±1.30b

87.17±0.12a 85.43±0.35b 83.60±0.13c 82.37±0.55d 80.94±1.12e

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hab

Water

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absorption index 2.23±0.14b 2.32±0.13b 2.62±0.08b 2.87±0.19b 3.86±0.56a

(WAI) (g/g)

Water solubility
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13.27±1.92a 13.07±1.69a 11.33±0.42a 6.21±0.69b 4.90±0.24b
(WS)
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Milk absorption

index (MAI) 1.56±0.08c 3.23±0.21b 3.59±0.25b 4.48±0.20a 4.63±0.34a

D

(g/g)
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Oil absorption

capacity (OAC) 1.52±0.03d 1.68±0.02cd 1.91±0.09c 2.38±0.09b 2.96±0.16a

EP

(g/g)
C

Emulsifying

49.51±4.44a 49.33±1.16a 46.05±2.01a 1.85±2.72b

AC

capacity (EC) 49.33±1.16ª

(%)

Emulsifying

stability (ES) 51.77±11.09b 59.83±6.20b 61.11±9.62b 17.17±0.86c 100±0.00a

(%)

Foam capacity 2.21±0.00ab 2.21±0.00ab 1.54±0.38bc 2.65±0.00a 1.32±0.00c

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(FC) (%)

Least gelation

capacity (LGC) 2% 2% 2% 2% 4%

(%)**

Means followed by the same letter in the line do not differ from each other, in

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agreement with the Tukey’s test at 5% significance. **minimum percentage where gel

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formation occurred.

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Table 6. Chemical composition of snack bars (SB)

Analyzes SB0 SB10 SB25 SB40 SB55

Moisture 8.42±0.06b 9.42±0.22a 9.55±0.04a 9.32±0.15a 8.80±0.27b

(g/100g)

Proteins 5.23±0.33a 5.24±0.16a 5.42±0.15a 5.72±0.10a 5.44±0.21a

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(g/100g)

1.59±0.09b 1.40±0.07b 1.72±0.15b 1.72±0.13b 2.64±0.25a

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Lipids

(g/100g)

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Ash (g/100g) 1.00±0.01c 1.09±0.02b 1.14±0.02b 1.14±0.04b 1.22±0.05a

Carbohydrate 79.43±0.36a 77.01±0.28b 75.16±0.21c 73.46±0.20d 70.27±0.22e

s
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TDF (g/100g) 4.02±0.18e 5.53±0.39d 6.67±0.26c 8.29±0.17b 11.30±0.21a
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SDF (g/100g) <0,1 <0,1 <0,1 <0,1 <0,1

IDF (g/100g) 4,02±0.18e 5,53±0.39d 6,67±0.26c 8,30±0.17b 11,30±0.21a

D

Energy2 352.92±0.99 341.63±0.53 337.82±1.1 332.23±0.42 326.58±1.2

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(kcal/100g) a 7c d
3e

Saturated Fat 0,37±0,00d 0,51±0,00c 1,09±0,00a 0,65±0,01b 1,14±0,04a

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(mg/100 g)
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Trans Fat n.d n.d n.d n.d n.d

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(mg/100 g)

Sodium 110,29±0,95 92,78±2,69a 93,20±3,86a 79,04±5,37b 79,46±4,88a

a b b b
(mg/100 g)

TDF: Total dietary fiber. IDF: Insoluble dietary fiber. SDF: Soluble dietary fiber. n.d.:

not detected. Means followed by the same letter in the line do not differ from each

other, in agreement with the Tukey’s test at 5% significance.

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Table 7. Physical parameters of snack bar formulations with 0, 10, 25, 40 and 55% of

corn bran

Physical parameters SB0 SB10 SB25 SB40 SB55

Specific
1.68±0,0b 1.89±0,0a 1.76±0,1ab 1.86±0,1a 1.71±0,1b
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volume(g/cm )

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Density(g/cm3) 0.54±0,0a 0.59±0,0a 0.57±0,0a 0.54±0,0a 0.58±0,0a

Hardness (N, n=10) 85.74±25,8ab 76.78±32,8b 89.12±44,3ab 98.78±45,0ab 126.26±42,3a

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Chromatic analysis

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(n=6)

L* 30,92±1,3b 31,49±1,3b 31,33±1,6b 32,03±1,3ab 33,97±0,9a

a* 2,11±0,3a
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1,81±0,1a 2,03±0,4a 1,87±0,1a 1,85±0,2a
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b* 8,53±0,6c 8,48±0,6c 8,90±0,5bc 9,67±0,8b 11,24±0,6a

8.79±0.59bc 9.15±0.49bc 9.86±0.82bc 11.40±0.60a

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C* 8.68±0.60c

hab 76.08±1.43c 77.88±0.94abc 77.06±3.05bc 78.97±1.13ab 80.65±0.87a

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Figure 1. Snack bar development flow chart

Dry ingredients Binding Ingredients Heating at 60 ºC

Mixture

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Cooking at 60 ºC for 5
minutes

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Lamination (1.0 cm)
and cut (9 cm x 2.5 cm)

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Figure 2. Scanning electron spectrographs at 2000x (upper) and 8000x (bottom) of the
five CB fractions

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- Corn bran, a by-product devalued, is an food ingredient with high nutritional value
- Antinutritional components were not found in corn bran
- Snack bars can be produced with corn bran

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