Accepted Manuscript

Addition of quinoa and amaranth flour in gluten-free breads: temporal profile and
instrumental analysis
Natlia Manzatti Machado Alencar, Caroline Joy Steel, Izabela Dutra Alvim, Elisa
Carvalho de Morais, Helena Maria Andre Bolini

PII:

S0023-6438(15)00132-2

DOI:

10.1016/j.lwt.2015.02.029

Reference:

YFSTL 4466

To appear in:

LWT - Food Science and Technology

Received Date: 16 October 2014

Revised Date:

19 February 2015

Accepted Date: 23 February 2015

Please cite this article as: Machado Alencar, N.M., Steel, C.J., Alvim, I.D., de Morais, E.C., Andre Bolini,
H.M., Addition of quinoa and amaranth flour in gluten-free breads: temporal profile and instrumental
analysis, LWT - Food Science and Technology (2015), doi: 10.1016/j.lwt.2015.02.029.

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ACCEPTED MANUSCRIPT
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Addition of quinoa and amaranth flour in gluten-free breads: temporal profile and
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instrumental analysis.

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Natlia Manzatti Machado Alencar *, Caroline Joy Steel , Izabela Dutra Alvim , Elisa Carvalho
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de Morais , Helena Maria Andre Bolini

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CEP: 13083-862, Brazil.

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Brazil.

School of Food Engineering, University of Campinas, Cidade Universitria Zeferino Vaz, Campinas, SP,

Technology Center of Grains and Chocolates (ITAL), Av. Brasil 2880, Campinas, SP, CEP: 1370-178,

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*Correspondence to: Natlia Manzatti Machado Alencar, Rua Monteiro Lobato 80, I.O.Box: 6121, CEP:

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13083-862, Campinas, SP, Brazil, Tel.: 55-19 -35214084; FAX: 55-19-35214060,

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natalia.manzatti@gmail.com

E-MAIL:

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Addition of quinoa and amaranth flour in gluten-free breads: temporal profile and
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instrumental analysis

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Abstract

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The objective of this study was to evaluate the influence of sweeteners and pseudocereals in

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gluten-free bread formulations. The quality parameters evaluated were specific volume, firmness, color,

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water activity, proximate composition, gross energy and an image analysis of the crumb. The sensory

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properties were analyzed using the time-intensity method. The bread containing amaranth, quinoa and

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sweeteners presented specific volume, firmness and water activity similar to those of the control bread,

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but showed higher protein, lipid and ash contents and a larger alveolar area. In the time-intensity analysis,

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those containing sweeteners did not differ statistically from the control bread (demerara sugar) for the

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sweet stimulus, but in relation to bitter stimulus, the bread containing quinoa and the sweeteners sucralose

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and sucralose-acessulfame showed higher maximum intensity. These results showed that it is possible to

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develop gluten-free breads with pseudocereals and sweeteners with similar sensory and physicochemical

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properties to those produced with starch-based formulations.

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Keywords: gluten-free bread, amaranth, quinoa, sweeteners, time-intensity analysis.

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1. Introduction

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The substitution of gluten is a great challenge and the majority of gluten-free breads available on

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the market is based on starches (Arent & Moore, 2006). Currently the gluten-free food manufacturers are

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investing in the use of whole grains including corn, rice, sorghum, buckwheat, amaranth and quinoa,

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since the majority of these are excellent fiber, iron and vitamin B sources (Thompson, 2009). The

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pseudocereals are considered as potentially gluten-free grains with an excellent nutrient profile, capable

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of diversifying this rising market (Alvarez-Jubete, Arendt & Gallagher, 2010a).

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The production of gluten-free breads has been widely studied recently (Cappa, Lucisan &

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Mariotti, 2013; Hera, Rosell & Gomez, 2014; Martnez, Daz & Gmez, 2014; Tsatsaragkou,

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Gounaropoulos & Mandala, 2014; Mohammadi et al., 2014). The choice of a product by consumers is

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determined by the interaction of non-sensory factors, as personal health in this case, and sensory factors

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(Jaeger, 2006). The time-intensity analysis allows one to dimension the sensory sensations perceived over

48

time, and the method provides information about flavor, odor and texture (Lawless & Heymann, 2010).

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The time-intensity method has been used for the last 25 years as an important tool because it allows

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comparison of the perception of sensory characteristics in a dynamic manner and can be applied to

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several food products with different objectives (Giovanni & Guinard, 2001).
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Some celiac patients develop diabetes lifelong, and in these cases they must consume not only

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gluten- but also sugar-free foods. Moreover, the increasing cases of obesity related to sugar intake have

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led to a greater need for studies with sugar substitutes, the sweeteners. The pseudocereals present as

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potential substitutes for gluten and they are sources of fiber. In this context, the objective of the present

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study was to evaluate the influence of sweeteners and pseudocereals in gluten-free bread, by way of a

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physicochemical analysis, and the time-intensity profile in relation to sweetness and bitterness.

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2. Materials and methods

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2.1.Materials
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Seven gluten and sucrose free loaf samples were prepared by partially substituting the mixture of

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starches by quinoa and amaranth flours, and the sucrose content by sweeteners. The ingredients used

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were: rice flour (Urbano , SP, Brazil), potato starch (Yoki , SP, Brazil), sea salt (Yoki , SP, Brazil),

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cassava starch (Amafil , PR, Brazil), sour tapioca starch (Hikari , SP, Brazil), amaranth and quinoa

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flours (R&S Blumos , SP, Brazil), demerara sugar (Native , SP, Brazil), dry yeast (Dr. oetker , SP,

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Brazil), xanthan gum (SweetMix , SP, Brazil), pasteurized liquid egg (Fleischmann , SP, Brazil), canola

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oil (Cargill , MG, Brazil), the sweeteners in powder form were: sucralose, stevia and

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sucralose/acessulfame-K blend (SweetMix , SP, Brazil) and water.

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2.2. Methods
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Formulations and loaves were developed in the Sensory Science and Consumer Study

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Laboratory of the School of Food Engineering, UNICAMP, Brazil. Subsequently the breads were

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produced in a food industry located in the Jundia city Brazil , called Grani Amici and specialized in

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manufacturing gluten-free loaves.

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2.2.1. Gluten-free breads manufacturing

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The concentrations of the ingredients water, canola oil, pasteurized liquid egg, xanthan gum, dry

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yeast and salt were kept constant for all the seven gluten-free bread samples. The concentrations of

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sweeteners, demerara sugar, quinoa flour, amaranth flour, sour tapioca starch, cassava starch, potato

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starch, and rice flour, varied according to the formulations (Table 1). These ingredients were obtained

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from local supermarkets in Campinas city, Brazil, or donated by suppliers and were all gluten-free.
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The concentrations of starch (rice flour, potato starch, cassava starch, sour tapioca starch,

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amaranth and quinoa whole flour) varied and was added until completing 100 g/100 g. The other

85

ingredients were added based on starch and flour content. The loaves were manufactured according to the

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steps indicated for mixing dry ingredients with liquid ingredients in an industrial mixer (Perfecta ) at

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medium speed for 3 min, until dough formation. Portions of 420 g of the dough were weighed into loaf

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tins (170 mm x 7 mm x 6 mm), placed in a proofing chamber (Perfecta ) at 35C for 20 min and

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subsequently baked at 195C in a rotary oven (Perfecta ) for 25 min, preheated to 195C. The loaves were

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cooled at room temperature. The loaves were then sliced in to 1 cm, slices packaged in transparent

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polyethylene bags, identified and stored frozen in a freezer until the day of evaluations. For loaves were

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not sliced, for specific volume evaluation.

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2.2.2. Samples preparation

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For samples presentation for assessors, samples were heated in an electric oven (Perfecta ) at

100C for 10 min. For the other analyses, the samples were thawed at room temperature.

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2.2.3. Physicochemical analyses

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The physicochemical analyses of gluten-free bread samples were performed at the Central

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Laboratory of the Food and Nutrition Department, Food Technology Department (UNICAMP/FEA) and

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Technology Center of Grains and Chocolates of Campinas (ITAL). Samples were evaluated in three

102

repetitions.

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2.2.3.1.Specific volume
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The specific volume of the loaves was determined according to AACC methodology (AACC,

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2000). The loaves were weighed in a semi-analytical balance and the volume measured by millet seed

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displacement. The specific volume was calculated from the relationship of volume/weight, and results

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were expressed as cm /g.

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2.2.3.2. Firmness
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The firmness of the crumb of gluten-free breads was evaluated according to AACC methodology

112

(AACC, 2000) using the TA-XT2 texture analyser and the program Dimension XTRA, Stable Micro

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Systems. The measurement of compression and force was carried out. The test was performed under the

114

following conditions: pre-test speed: 1.0 mm/s, test speed: 1.7 mm/s, and post-test speed: 10.00 mm/s,

115

compression 40%. A cylindrical aluminum probe of 36 mm diameter (P36/R) was used.

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2.2.3.3. Color

The crumb color of gluten-free breads was determined using the CIELab system, evaluating the
119

color parameters L* (luminosity), a* (green-red) and b* (blue-yellow) in a Hunter Lab model Color Quest

120

II spectrophotometer (Hunter Associates Laboratory, Reston, VA, USA), The apparatus was calibrated

121

with the illuminant D65, 10 hue angle and the RSIN calibration mode (Minolta, 1994).

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123

2.2.3.4. Water activity

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The water activity of the crumb of gluten-free bread samples was determined using the Aqualab

analyzer (Decagon, Brazil).

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127

2.2.5. Proximate composition

128

The analyses of proximate composition of gluten-free bread samples were carried out in order to

129

characterize the bread samples. The moisture content, crude protein content, ash content and ether extract

130

were determined using the AACC (2000) methodologies. The carbohydrates were calculated by

131

difference.

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2.2.3.6. Gross energy

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135

The gross energy (kJ/g) of the samples was obtained using a Parr calorimetric pump and convert

in kcal/g.

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137

2.2.3.7. Analysis of the bread crumb images

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Digital images of 1 cm thick slices of gluten-free bread samples were captured in the jpeg

139

format. The colored images were first converted to the gray scale using the program Photo Filter, and

140

then analyzed by the software Image J. The values for the digitalized images were obtained in pixels,

141

converted into centimeters using bars of known length (355 pixels equivalent to 1 cm), and the mean area

142

of the alveolus and the number of alveoli per cm was then determined.

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2.2.3.8. Statistical analyses

145

The results of the physicochemical analyses were evaluated using univariate statistical analysis

146

(ANOVA), and the means compared by Tukeys test at 5% of significance. The calculations were

147

performed using the Statistical Software Analysis System SAS (SAS Institute, 2009).

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2.3. Time-intensity analysis

150

Time-intensity analyses were carried out in individual air-conditioned (22 C) booths with white

151

light. Water was provided for palate cleansing. Sessions were held at the Laboratory of Sensory Science

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and Consumer Study of the Food and Nutrition Department (School of Food Engineering/University of

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Campinas) and samples (a half of a slice of bread including crumb and crust) were presented in white

154

disposable plates with 3-digit numbers randomly coded.

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gluten-free bread samples. The two attributes were evaluated in separate instances.
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The time-intensity analysis was performed for the sweet taste and bitter taste stimuli of the

Approval for the study was obtained from the Ethics Committee of the University of Campinas,

and all volunteers gave a written consent.

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2.3.1. Selection and training of assessors

161

A preselection of candidates was carried out using Walds sequential analysis (Meilgaard, Civille

162

& Carr, 2004). Two commercial gluten-free bread samples were used and previously tested to obtain a

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1% of significant difference level. Triangle difference tests were applied with 28 consumers using these

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gluten-free bread samples (Gomes et al., 2014). After the preselection, fourteen assessors were chosen.
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All the assessors determined the references by a consensus, and they were then trained with

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respect to the product attributes using identified references (Table 2). The direct contact of the individuals

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with the reference of maximum intensity for each stimulus (sweet taste and bitter taste) led to training for

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the formation of sensory memory and equalization among assessors. The panel was trained in six 1 hour

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training sessions, in which each assessor was trained with respect to the program and movement of the

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mouse in the analyses.

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2.3.2. Selection of the assessor and evaluation of attributes by time-intensity analysis

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In the selection of the assessors, the analysis of variance (ANOVA) was applied for each panelist

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and each parameter in each stimulus separately, and 12 assessors of 14 were selected to participate based

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on their discriminative power, repeatability and agreement with the team Damsio & Costell (1991),

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verified by their discriminating capability (p<0.50) and repeatability (p>0.05), while an individual

177

consensus was also considered. For this selection, the assessors evaluated the attributes by way of a

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monadic presentation with four repetitions, registering the intensity of the attribute on a structured linear

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scale from zero to nine (0=none, 4.5=moderate, and 9=strong) on the computer screen, as a function of

180

the time lapsed.

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On hearing the first signal emitted by the computer, the assessor put the whole sample in to the

mouth and, using the mouse, registered the intensity of the particular sensory attribute on the scale. On

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the second signal, the assessor swallowed the sample and the third signal indicated the end of the test. The

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time parameters used in the stimulus evaluation steps, registered on a 9 cm scale were: 10 s of initial

185

waiting, 30 s of residence in the mouth and 60 s after swallowing.

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The 12 assessors evaluated the samples by way of a balanced complete block design (Macfie,

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1989), in a monadic way with three repetitions. Data collection for the time-intensity analysis were

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carried out on a computer using the software Time Intensity Analysis of Food and Tastes (TIAF)

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(Universidade Estadual de Campinas, 2012).

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The curve data obtained were keyed in on the spreadsheet of the Excel for Windows program and

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analyzed by the SAS statistical program (SAS Institute, 2009). The curve parameters evaluated were:

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Imax maximum stimulus intensity recorded by the assessor; Tmax time when the maximum intensity

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was recorded by the assessor; Area total area of the time curve x intensity; Ttot total duration time of

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the stimulus. The parameters were evaluated using the analysis of variance (ANOVA) and Tukeys means

195

test.

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3. Results and discussion

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3.1. Physicochemical analyses

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Tables 3 and 4 show the mean values obtained in the physicochemical analyses and image

analyses of the gluten-free bread samples.

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3.1.1. Specific volume

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The loaves weighed approximately 220 g and showed a specific volume varying from 2.30 to

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2.88 cm/g, as described in Table 3. Low values were observed for this parameter. The sample containing

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quinoa flour and the sweetener stevia presented the highest mean for specific volume, but was not

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statistically different (p<0.05) from the control, amaranth and sucralose, amaranth and sucralose-

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acessulfameK, quinoa and sucralose-acessulfame-K loaves.

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With the exception of the loaves containing amaranth flour and the sweetener stevia, the specific

210

volumes of the other samples did not vary significantly (p<0.05) from the value obtained for the control.

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A similar result was found by Sciarini et al. (2012), who tested different hydrocolloids in gluten-free

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bread, and found no statistical difference between the specific volume of the loaves made using xanthan

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gum and the control loaves (with no addition of hydrocolloid). This parameter is important for acceptance

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by consumers, because loaves with higher specific volume are usually more preferred.

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3.1.2. Firmness
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The gluten-free breads evaluated in this study had a starch-rich base formulation, and the

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alterations in the starch granules during baking determined the crumb structure. Thus, heating causes an

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extension of the amylopectin crystals, resulting in swelling of the granules and alterations in the textural

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characteristics (Patel, Waniska & Seetharaman, 2005). In addition, egg is considered to be an important

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ingredient in gluten-free bread formulations, since it provides softer crumbs when compared to those

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formulated without egg, the latter being firmer and less springy (Milde, Ramallo & Puppo, 2012). The

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results for firmness varied from 259.33 to 568.97 g. It can be seen in Table 3 that the sample with quinoa

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and stevia was characterized by presenting a lower firmness, differing statistically (p < 0.05) from the

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other samples. However, the sample with quinoa and sucralose presented the higher mean of firmness and

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did not differ statistically (p > 0.05) from the control and amaranth and sucralose-acessulfame-K

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samples. According to Esteller et al. (2004), the hardness or firmness of the bread is related to the applied

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force that causes rupture or deformation of the samples, and is correlated with the human bite. Hager &

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Arent (2013) showed that xanthan gum strengthened the crumb structure of gluten-free bread, that is,

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bread supplemented with xanthan gum showed an increase in hardness.

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3.1.3. Color

The color of bread is considered to be a very important factor in product commercialization, and
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is directly influenced by the ingredients making up the formulation and by the baking conditions (Silva et

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al., 2009).

The instrumental color analysis as described in Table 3 showed that the samples presented means
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varying from 79.00 - 72.05 for the parameter L*, and were therefore characterized by a lighter color. This

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result was expected since gluten-free bread formulations were produced from rice flour and other starches

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and light quinoa and amaranth flours. Matos & Rosell (2013) found high values for L*, varying from

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62.24 81.50 in gluten-free bread based on rice flour. All the loaves showed low mean values for

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parameter a* and hence presented little reddish coloration. The samples containing the sweeteners stevia

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and sucralose-acessulfame-K and those containing quinoa flour and sucralose were characterized by

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higher mean values for parameter b*, statistically different (p < 0.05) from the control loaves and from

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those containing amaranth flour and sucralose. Thus, these samples were more intensely yellow. The

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mean values found for the parameters a* and b* in the present study are similar to those found for other

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gluten-free breads containing pseudocereals (buckwheat, amaranth, quinoa) published in the literature,

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which were characterized by yellowing of the crumb as compared to the control bread (Wronkowska,

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Haros & Soral-mietana, 2013; Alvarez-Jubete et al., 2010b).

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3.1.4. Water activity

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According to the results presented in Table 3, the water activity varied from 0.90 to 0.92 in the

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gluten-free bread samples evaluated. The sample produced with the quinoa flour and the sweetener

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sucralose stood out for having the lowest mean value (0.900.01) being statistically different (p < 0.05)

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from the gluten-free bread sample with quinoa and sucralose. Mariotti, Pagani & Lucisano, (2013) found

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values of about 0.99 to 0.98 for the water activity of gluten-free bread containing 40 g/100 g of

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buckwheat flour in its composition.

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3.1.5. Gross energy

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The majority of the energy in bread comes from the starches. One hundred grams of white bread

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provide 219 kcal, whilst the same weight of whole wheat bread provides 217 kcal (OConnor, 2012). The

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caloric values of the breads formulated in the present study are presented in Table 3, and varied from

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231.43 to 261.91 kcal considering 100 g of gluten-free bread samples. The bread samples formulated with

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quinoa flour and the sweeteners sucralose-acesulfame-K presented the lowest caloric indices, and

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differed significantly (p < 0.05) from the other samples.

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3.1.6. Proximate composition

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Table 4 shows that gluten-free breads formulated with pseudocereals flours and sweeteners

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presented higher ash contents, differing statistically (p < 0.05) from the control sample. In relation to the

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lipid contents, the gluten-free bread with quinoa and sucralose-acesulfame-K presented the highest mean

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value, which did not differ statistically (p > 0.05) from the bread with amaranth and stevia. The control

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bread presented the lowest mean value for this parameter, and did not differ significantly (p > 0.05) from

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the breads with quinoa and sucralose, quinoa and stevia, and amaranth and sucralose-acesulfame-K.

1
0

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According to Sanz-Penella (2013), the incorporation of 10 g/100 g or more amaranth flour into bread

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formulations significantly increases the protein, lipid and ash contents and decreases the starch content.

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The highest protein contents were found in all the samples produced with amaranth flour and in the bread

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formulated with quinoa flour and the sweetener sucralose-acesulfame-K. Recently, Segura & Rosell

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(2011) presented the nutritional composition of 11 types of gluten-free breads, which contained 0.91-

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1.05% protein, 2.00-26.10% lipids, 1.10-5.43% minerals and 68.42-92.96 carbohydrates, showing a

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similar nutritional profile to that found in the current study. It can be seen that the bread samples

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presented 42.8 49.89% of carbohydrate, since the gluten-free breads are based mainly on carbohydrates.

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In general, the principal components in the formulations are flours and starches (Matos & Rosell, 2012).

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3.1.7. Image analyses

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An analysis of the images of the crumb structures of gluten-free bread samples (Figure 1 and

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Table 4) indicates that the formulations with amaranth and quinoa flours presented a smaller number of

286

alveoli but with larger areas, being oposite to the control gluten-free bread. These results also can be seen

287

in Table 4, in which the number of alveoli presented the highest mean for the control sample, while the

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lowest mean value was found for this sample in relation to the total alveolar area.
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The use of buckwheat, amaranth and quinoa flours provides a structure with larger alveoli in the

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bread as compared to gluten-free bread with a starch based formulation (Alvarez-Jubete, Arendt &

291

Gallagher, 2010a). A great variety in structure can be found amongst the gluten-free breads, some

292

presenting a larger alveolar area and others a larger number of alveoli. This fact is due to the diversity of

293

ingredients added to the dough (Matos & Rosell, 2013).

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3.2. Time-intensity analysis

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3.2.1. Time-intensity analysis for sweet taste stimulus of gluten-free breads

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Table 5 shows the mean values for sweetness obtained for the gluten-free bread samples in

299

relation to each parameter of the curve. It was possible to observe that no significant difference (p > 0.05)

300

was found among samples in relation to the evaluated parameters: Imax, Tmax, Ttotal, and Area. The

301

control gluten-free bread sample, which was produced with demerara sugar, presented mean values for

302

maximum intensity of sweetness statistically equal to the samples using sweeteners as substitutes of

303

sugar. Thus, the replacement of sugar by sweetener agents did not alter the perception of sweetness

304

intensity for the gluten-free breads.

305

Figure 2 represents the profile trend for the sweet stimulus as registered by the assessors. Time-

306

intensity curves showed that the seven gluten-free bread samples added with quinoa, amaranth and

307

sweeteners presented similar temporal profiles. The similarities between the time-intensity curves

308

obtained for the sweeteners sucralose, stevia and sucralose-acessulfame-K showed their potential to

309

substitute sucrose. Morais, Cruz & Bolini (2013) evaluated the time-intensity profile of gluten-free breads

310

in which the sugar was substituted by prebiotics and sweeteners. It was shown that the assessors

311

perceived a more intense sweet taste in the breads containing demerara sugar, followed by those

312

sweetened with fructooligosaccharides, which showed a similar behavior for the perception of sweetness

313

throughout the test.

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3.2.2. Time-intensity analysis for bitter taste stimulus of gluten-free breads

316

Table 5 shows the mean values for bitterness obtained for the gluten-free bread samples in

317

relation to each parameter of the time-intensity curve registered for this stimulus, and Figure 3 represents

318

the profile trend for the bitter stimulus as registered by the assessors. No significant difference (p > 0.05)

319

was observed between the samples for the maximum intensity time (Tmax). A significant difference (p <

320

0.05) was observed among samples in relation to Imax. The sample developed with quinoa and sucralose-

321

acesulfame-K presented a higher mean value for intensity of bitterness followed by the samples with

322

quinoa and sucralose and quinoa and stevia. This result may relate to the addition of quinoa flour with

323

perceived increasing on bitterness intensity. On the other hand, the control sample presented a lower

324

mean value for intensity of bitterness, as can be seen in Figure 3, and did not differ statistically (p > 0.05)

325

from the samples with amaranth and sweeteners. These data may be observed using the time-intensity

326

curves, which show how similar their profiles are in the behavior of bitterness perception.

327
328

3.2.3. Bitterness and sweetness temporal profile in gluten and sucrose-free breads
329

An analysis comparing the time-intensity profile of each sample in relation to bitterness and

330

sweetness allowed evaluating the samples individually in relation to the stimuli simultaneously, on the

331

same graph. It can be seen, in Figure 4, that in exception of the breads containing quinoa, sucralose and

332

sucralose-acessulfame-K, the sweet stimulus was perceived with greater intensity by the assessors.

333

However, the use of the sweeteners showed a greater duration of the stimulus, suggesting a residual

334

sweetness for these samples. Differently, the bread samples containing quinoa, sucralose and sucralose-

335

acessulfame-K were perceived as having a greater bitter taste.

336

Figure 4 shows that the samples which presented the more similar temporal profiles in relation to

337

bitterness and sweetness when compared with the control sample were the samples with amaranth and the

338

sweeteners. These results show that for gluten-free breads produced with sugar replacement by

339

sweeteners and partial replacement of starch by pseudocereals, the addition of amaranth in formulations

340

with sweeteners led to a more similar temporal sensory profile in relation to a conventional one (with

341

demerara sugar and starch based), than the addition of quinoa.

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343

4. CONCLUSIONS

344
The bread samples containing pseudocereals and sweeteners presented values for specific
346

volume, firmness and water activity similar to those of the control formulation. It can be seen that the

347

bread samples containing amaranth and quinoa presented greater amounts of proteins, lipids and ash,

348

improving their nutritional profile.

The bread samples developed with demerara sugar showed a similar behavior with respect to the

350

perception of sweetness to those developed with sweeteners. With respect to the perception of bitter taste,

351

the bread samples containing quinoa and the sweeteners sucralose, sucralose-acessulfame-K and stevia

352

presented a greater intensity for this stimulus. However, in the analysis of the multiple time-intensity

353

profile, sweetness was perceived with greater intensity in most samples by the assessors.
The use of amaranth, quinoa and sweeteners was shown to be efficient in the development of

355

gluten-free breads. This research opens up new opportunities for the gluten- and sucrose-free bakery

356

industry, showing possibilities in developing gluten-free breads for a group of the population with special

357

needs, as celiacs, gluten intolerants, diabetics, and/or both of them.

358
359

Acknowledgments

360

The present study was carried out with the support of the Brazilian National Research Council (CNPq).

361
362

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% Base flour

Starch

Table 1. Ingredients of gluten and sugar free bread formulations.

Ingredients
F1
F2
F3
F4
F5
F6
F7
(g/100g (g/100g (g/100g (g/100g (g/100g (g/100g (g/100g)
)
)
)
)
)
)
Rice flour
61.64
49.32
49.32
49.32
49.32
49.32
49.32
Potat
13.68
10.95
10.95
10.95
10.95
10.95
10.95
o
starc
20.56
16.43
16.43
16.43
16.43
16.43
16.43
hstarch
Sour
4.12
3.30
3.30
3.30
3.30
3.30
3.30
tapioca
starch
0
20
0
20
0
20
0
Amara
nth
whole
0
0
20
0
20
0
20
flour
whole
Quinoa
flour
100
100
100
100
100
100
100
Amount
of
2.73
0
0
0
0
0
0
Demer
ara
0
0.003
0.003
0
0
0.002
0.002
sugar
Sucral
Stevia
0
0
0
0.01
0.01
0
0
Acesulfa
0
0
0
0
0
0.005
0.005
me- K
Salt
2.05
2.05
2.05
2.05
2.05
2.05
2.05
Dry yeast
2.05
2.05
2.05
2.05
2.05
2.05
2.05
Xanthan
0.35
0.35
0.35
0.35
0.35
0.35
0.35
gum
Pasteuriz
37.67
37.67
37.67
37.67
37.67
37.67
37.67
ed
liquid
10.28
10.28
10.28
10.28
10.28 10.28
10.28
egg
Canola
Wate
82.25
82.25
82.25
82.25
82.25 82.25
82.25
r at
4C F2: amaranth sucralose; F3: quinoa sucralose; F4: amaranth stevia; F5: quinoa stevia;
Note: F1: control;
F6: amaranth sucralose/acessulfame-K; F7: quinoa sucralose/acesulfame-K.

Table 2. Definitions and references for the stimuli evaluated by assessors for gluten- and sugar-free bread
in time intensity analysis.
Attributes

Definition

Reference

Sweetness

Characteristic taste of sucrose

solution.

None sweetness: Biscuit with fibers Triunfo (SP,

Brazil), less salt water type.
A lot sweetness: Toast lightly sweet Bauduco (SP,
Brazil).

Bitter taste

Characteristic taste of caffeine

solution.

None bitterness: Biscuit with fibers Triunfo (SP,

Brazil), less salt water type.
Very bitterness: Bread without gluten with 40 g/100g of
amaranth. Ingredients mixture: 108 g of rice flour
Urbano (SP, Brazil); 24 g of potato starch Yoki (SP,
Brazil), 36 g of cassava flour Yoki (SP, Brazil), 7.2 g
sour tapioca flour Hikari ( SP, Brazil),116.8 g of
amaranth flour R&S Blumos (SP, Brazil), 8 g of raw
sugar Native (SP, Brazil), 2 eggs, 30ml oil canola
Cargil (MG, Brazil), 240 ml de water, 6 g salt Yoki
(SP, Brazil) and 6 g dry yeast Dr. Ockter (SP, Brazil). It
was fermented at 35 C during 20 minutes and baked in
electric oven during 20 minutes at 220C temperature.

ACCEPTED MANUSCRIPT

Table 3. Specific volume, firmness, color (L*, a*, b*), water activity, and energy averages of gluten and sugar free bread samples.
Samples

Specific Volume

Firmness

(cm/g)

(g)

abc

0.02

abc

Control

2.55

Amaranth
sucralose
Quinoa sucralose

2.58

abc

0.07

470.89
76.86
c
396.01 13.60

2.36

bc

0.06

568.97 85.15

Amaranth stevia

2.30 0.07

Quinoa stevia

2.88 0.29

394.76 7.33
d

259.33 11.53

abc

0.09

546.36 67.89

ab

0.03

447.89 43.83

Amaranth
sucraloseacesulfame-K

2.65

Quinoa sucraloseacesulfame-K

2.67

ab

bc

L*

a*

b*

79.00 1.54

0.29 0.11

19.16 0.35

76.98
ab
1.78
abc
76.51 1.28

1.08 0.42

21.91 1.78

bc

21.91 1.01

21.65 1.29

bc

22.38 0.95

24.20 0.79

ab

24.37 1.31

0.47 0.20

75.25 1.33

bcd

1.12 0.11

73.43 1.47

cd

0.63 0.27

74.13 0.80

cd

1.91 0.42

1.18 0.36

72.05 1.23

b
b

ab
ab
a
a

Gross energy

Wat
er

kcal/100g

ab

0.91 0.0
0
ab
0.91 0.0
0
0.92
a
0.00
ab
0.91 0.0
1
ab
0.91 0.0
0
ab
0.91 0.0

243.12 2.31
bc

248.19 1.95
d

231.53 1.81

0.90 0.01

Means with a same superscript letter in the same column are not significantly different at a 5% level according to Tukeys test (p < 0.05).

bc

248.85 1.06
a

261.91 2.02
b

254.17 0.01

225.98 1.80

ACCEPTED MANUSCRIPT

Table 4. Proximate composition averages and averages of image analyses of gluten- and sugar-free bread samples.
Sample
Cont
rol
Amaranth sucralose

Moisture (%)
ab

43.94 2.03

Ash (%)
c

1.26 1.27

41.08

ab

1.27

1.65 1.65

Quinoa sucralose

45.04

ab

3.94

1.45 1.45

Amaranth stevia

41.51 4.30

ab

1.61 1.60

Quinoa stevia

44.94 4.69

1.47 1.47

Amaranth sucraloseacesulfame-K

43.91 2.03

1.53 1.53

Quinoa sucraloseacesulfame-K

35.38 1.75

1.67 1.67

Lipid (%)
d

6.55 0.12

7.68 0.11

7.01 0.04

ab

7.98 0.41

6.37 0.08

ab

ab

Protein (%)
c

3.44 0.13

bc

4.14 0.08

cd

3.70 0.06

4.34 0.24

bcd

7.11

0.43

8.69 0.56

Carbohydrate * (%)

Number of alveoli/cm

44.81

67.79

1.99

ab

45.45

57.35

bc

42.80

48.03

44.56

56.86

bc

43.52

42.54

43.48

61.02

49.89

50.50

3.70 0.09
abc

3.97

0.18

4.37 0.34

Chemical composition results were expressed in based moist, carbohydrate were calculated by difference.
Means with a same superscript letter in the same column are not significantly different at a 5% level according to Tukeys test (p < 0.05).

Total area alveoli mm/cm

acd

ce

abc

ab

bde

bcde

2.66

ab

3.35

bcde

2.79

4.25

bcde

2.13

ab

3.30

ACCEPTED MANUSCRIPT
Table 5. Averages of the parameters obtained from time-intensity curves for the sweetness and bitterness
stimulus from gluten- and sugar-free bread samples.
Samples

Imax

Timax

Ttotal

Area

Sweetness
a

25.91

4.92

28.25

Quinoa stevia

5.15

28.55

Amaranth sucralose

4.60

27.14

Quinoa sucralose

4.84

26.09

Amaranth sucralose acesulfame-K

4.81

27.68

Quinoa sucralose-acesulfame-K

4.85

MDS

0.84

Control

5.01

Amaranth stevia

43.11

136.35

45.06

141.66

45.98

148.51

42.78

122.67

46.37

142.97

44.38

131.99

27.26

46.37

131.75

4.28

5.51

32.07

Bitter
d

25.92

cd

26.65

bc

25.68

cd

27.41

ab

26.94

cd

25.96

0.96

Control

3.245

Amaranth stevia

4.37

Quinoa stevia

4.32

Amaranth sucralose

3.92

Quinoa sucralose

5.03

Amaranth sucralose acesulfame-K

4.06

Quinoa sucralose-acesulfame-K

5.40

MDS

37.67

86.19

39.85

41.39

39.58

41.01

39.06

28.32

45.23

4.98

6.46

ab

115.91

abc

ab

115.13

abc

ab

102.16

bc

ab

134.18

ab

ab

102.48

bc

147.50

35.81

Note: Means with a same superscript letter in the same column are not significantly different at a 5%
level. Imax: maximum intensity recorded by the assessor; Timax: time in which the maximum
intensity was recorded; Ttot: total duration time of the stimulus; Area: area of the time curve x
intensity. Unstructured linear scale of 9 cm anchored with the words less on the left and very on
the right. Averages with a same superscript letter in the column are not significantly different at a 5%
level. MDS: minimum significant difference in Tukeys test * p < 0.05.

2
2

U
N

M
D

Fig. 1. Digital images of gluten- and sugar-free bread crumbs. A Control; B Amaranth and sucralose;
C Quinoa and sucralose; D Amaranth and stevia; E Quinoa and stevia, F Amaranth and
sucralose/acesulfameK; G Quinoa and sucralose/acesulfameK. Expansion of 20X for all images.

Intensity

8
7
6
5
4
3
2
1
0

10

20

T
P
I
R
C
S
U
N
A
M
30

40

50

60

Time(s)

Control
Quinoa stevia Quinoa Sucralose
Amaranth sucralose/acesulfame-K

Amaranth stevia
Amaranth sucralose
Quinoa sucralose/acesulfame-K

Fig. 2. Time-intensity curves and characteristics of sweetness stimulus for gluten and sugar free bread
samples.

Intensity

8
7
6
5
4
3
2
1
0

10

20

30

Time (s)

Control
Quinoa stevia Quinoa Sucralose
Amaranth sucralose/acesulfame-K

T
P
I
R
SC

40

U
N

50

60

Amaranth stevia
Amaranth sucralose
Quinoa sucralose/acesulfame-K

Fig. 3. Time-intensity curves and characteristics of bitter stimulus for gluten and sugar free bread
samples.

C
C

T
EP

N
A
M
ED

T
P
I
R
C
S
U

Fig. 4. Time-intensity profile of gluten and sugar free bread samples for sweetness and bitterness stimuli.

Highlights

2
3

The pseudocereals increase the nutritional quality of gluten-free breads.

The sweetness did not differ statistically between the gluten-free bread

5
6

samples.

8
9

The addition of quinoa increase the bitterness intensity in gluten-free

breads.

10

11

Pseudocereals and sweeteners presented a satisfactory performance in

12

bread.