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Addition of quinoa and amaranth

flour in gluten-free breads: temporal
profile and instrumental analysis
Helena Bolini

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 LWT - Food Science and Technology 62 (2015) 1011e1018

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LWT - Food Science and Technology

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

Addition of quinoa and amaranth flour in gluten-free breads:

Temporal profile and instrumental analysis
Nata
lia Manzatti Machado Alencar a, *, Caroline Joy Steel a, Izabela Dutra Alvim b,
Elisa Carvalho de Morais a, Helena Maria Andre Bolini a
a
ria Zeferino Vaz, Campinas, SP, CEP 13083-862, Brazil
School of Food Engineering, University of Campinas, Cidade Universita
b
Technology Center of Grains and Chocolates (ITAL), Av. Brasil 2880, Campinas, SP, CEP 1370-178, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: The objective of this study was to evaluate the influence of sweeteners and pseudocereals in gluten-free
Received 16 October 2014 bread formulations. The quality parameters evaluated were specific volume, firmness, color, water ac-
Received in revised form tivity, proximate composition, gross energy and an image analysis of the crumb. The sensory properties
19 February 2015
were analyzed using the time-intensity method. The bread containing amaranth, quinoa and sweeteners
Accepted 23 February 2015
Available online 4 March 2015
presented specific volume, firmness and water activity similar to those of the control bread, but showed
higher protein, lipid and ash contents and a larger alveolar area. In the time-intensity analysis, those
containing sweeteners did not differ statistically from the control bread (demerara sugar) for the sweet
Keywords:
Gluten-free bread
stimulus, but in relation to bitter stimulus, the bread containing quinoa and the sweeteners sucralose and
Amaranth sucralose-acesulfame showed higher maximum intensity. These results showed that it is possible to
Quinoa develop gluten-free breads with pseudocereals and sweeteners with similar sensory and physico-
Sweeteners chemical properties to those produced with starch-based formulations.
Time-intensity analysis © 2015 Elsevier Ltd. All rights reserved.

1. Introduction
The substitution of gluten is a great challenge and the majority
of gluten-free breads available on the market is based on starches
(Arent & Moore, 2006). Currently the gluten-free food manufac-
turers are investing in the use of whole grains including corn, rice,
sorghum, buckwheat, amaranth and quinoa, since the majority of
these are excellent fiber, iron and vitamin B sources (Thompson,
2009). The pseudocereals are considered as potentially gluten-
free grains with an excellent nutrient profile, capable of diversi-
fying this rising market (Alvarez-Jubete, Arendt, & Gallagher, 2010).
The production of gluten-free breads has been widely studied
recently (Cappa, Lucisano, & Mariotti, 2013; Hera, Rosell, & Gomez,
2014; Martínez, Díaz, & Go
mez, 2014; Mohammadi, Sadeghniaa,
Azizi, Neyestani, & Mortazavian, 2014; Tsatsaragkou,
Gounaropoulos, & Mandala, 2014). The choice of a product by
consumers is determined by the interaction of non-sensory factors,
as personal health in this case, and sensory factors (Jaeger, 2006).
The time-intensity analysis allows one to dimension the sensory
sensations perceived over time, and the method provides infor-
mation about flavor, odor and texture (Lawless & Heymann, 2010).
The time-intensity method has been used for the last 25 years as an
important tool because it allows comparison of the perception of
sensory characteristics in a dynamic manner and can be applied to
several food products with different objectives (Giovanni &
Guinard, 2001).
Some celiac patients develop diabetes lifelong, and in these cases
they must consume not only gluten- but also sugar-free foods.
Moreover, the increasing cases of obesity related to sugar intake have
led to a greater need for studies with sugar substitutes, the sweet-
eners. The pseudocereals present as potential substitutes for gluten
and they are sources of fiber. In this context, the objective of the
present study was to evaluate the influence of sweeteners and pseu-
docereals in gluten-free bread, by way of a physicochemical analysis,
and the time-intensity profile in relation to sweetness and bitterness.
2. Materials and methods

2.1. Materials
* Corresponding author. Rua Monteiro Lobato 80, I.O. Box 6121, Campinas, SP, CEP
13083-862, Brazil. Tel.: þ55 19 35214084; fax: þ55 19 35214060. Seven gluten and sucrose free loaf samples were prepared by
E-mail address: natalia.manzatti@gmail.com (N.M. Machado Alencar). partially substituting the mixture of starches by quinoa and

http://dx.doi.org/10.1016/j.lwt.2015.02.029
0023-6438/© 2015 Elsevier Ltd. All rights reserved.
1012 N.M. Machado Alencar et al. / LWT - Food Science and Technology 62 (2015) 1011e1018

amaranth flours, and the sucrose content by sweeteners. The in-
gredients used were: rice flour (Urbano®, SP, Brazil), potato starch
(Yoki®, SP, Brazil), sea salt (Yoki®, SP, Brazil), cassava starch (Ama-
fil®, PR, Brazil), sour tapioca starch (Hikari®, SP, Brazil), amaranth
and quinoa flours (R&S Blumos®, SP, Brazil), demerara sugar
(Native®, SP, Brazil), dry yeast (Dr. oetker®, SP, Brazil), xanthan gum
(SweetMix®, SP, Brazil), pasteurized liquid egg (Fleischmann®, SP,
Brazil), canola oil (Cargill®, MG, Brazil), the sweeteners in powder
form were: sucralose, stevia and sucralose/acesulfame-K blend
(SweetMix®, SP, Brazil) and water.
2.2. Methods
Formulations and loaves were developed in the Sensory Sci-
ence and Consumer Study Laboratory of the School of Food En-
gineering, UNICAMP, Brazil. Subsequently the breads were
produced in a food industry located in the Jundiaí city e Brazil,
called “Grani Amici” and specialized in manufacturing gluten-free
loaves.
2.2.1. Gluten-free breads manufacturing
The concentrations of the ingredients water, canola oil,
pasteurized liquid egg, xanthan gum, dry yeast and salt were kept
constant for all the seven gluten-free bread samples. The concen-
trations of sweeteners, demerara sugar, quinoa flour, amaranth
flour, sour tapioca starch, cassava starch, potato starch, and rice
flour, varied according to the formulations (Table 1). These in-
gredients were obtained from local supermarkets in Campinas city,
Brazil, or donated by suppliers and were all gluten-free.
The concentrations of starch (rice flour, potato starch, cassava
starch, sour tapioca starch, amaranth and quinoa whole flour)
varied and was added until completing 100 g/100 g. The other in-
gredients were added based on starch and flour content. The loaves
were manufactured according to the steps indicated for mixing dry
ingredients with liquid ingredients in an industrial mixer
(Perfecta®) at medium speed for 3 min, until dough formation.
Portions of 420 g of the dough were weighed into loaf tins
(170 mm  7 mm  6 mm), placed in a proofing chamber
(Perfecta®) at 35  C for 20 min and subsequently baked at 195  C in
a rotary oven (Perfecta®) for 25 min, preheated to 195  C. The loaves
were cooled at room temperature. The loaves were then sliced in to
1 cm, slices packaged in transparent polyethylene bags, identified
and stored frozen in a freezer until the day of evaluations. For loaves
were not sliced, for specific volume evaluation.
2.2.2. Samples preparation
For samples presentation for assessors, samples were heated in
an electric oven (Perfecta®) at 100  C for 10 min. For the other
analyses, the samples were thawed at room temperature.
2.2.3. Physicochemical analyses
The physicochemical analyses of gluten-free bread samples
were performed at the Central Laboratory of the Food and Nutrition
Department, Food Technology Department (UNICAMP/FEA) and
Technology Center of Grains and Chocolates of Campinas (ITAL).
Samples were evaluated in three repetitions.
2.2.3.1. Specific volume. The specific volume of the loaves was
determined according to AACC methodology (AACC, 2000). The
loaves were weighed in a semi-analytical balance and the volume
measured by millet seed displacement. The specific volume was
calculated from the relationship of volume/weight, and results
were expressed as cm3/g.
2.2.3.2. Firmness. The firmness of the crumb of gluten-free breads
was evaluated according to AACC methodology (AACC, 2000) using
the TA-XT2 texture analyser and the program Dimension XTRA,
Stable Micro Systems. The measurement of compression and force
was carried out. The test was performed under the following con-
ditions: pre-test speed: 1.0 mm/s, test speed: 1.7 mm/s, and post-
test speed: 10.00 mm/s, compression 40%. A cylindrical aluminum
probe of 36 mm diameter (P36/R) was used.
2.2.3.3. Color. The crumb color of gluten-free breads was deter-
mined using the CIELab system, evaluating the color parameters L*
(luminosity), a* (green-red) and b* (blue-yellow) in a Hunter Lab
model Color Quest II spectrophotometer (Hunter Associates Labo-
ratory, Reston, VA, USA), The apparatus was calibrated with the
illuminant D65, 10 hue angle and the RSIN calibration mode
(Minolta, 1994).
2.2.3.4. Water activity. The water activity of the crumb of gluten-
free bread samples was determined using the Aqualab analyzer
(Decagon, Brazil).
2.2.4. Proximate composition
The analyses of proximate composition of gluten-free bread
samples were carried out in order to characterize the bread sam-
ples. The moisture content, crude protein content, ash content and

Table 1
Ingredients of gluten and sugar free bread formulations.

Ingredients F1 (g/100 g) F2 (g/100 g) F3 (g/100 g) F4 (g/100 g) F5 (g/100 g) F6 (g/100 g) F7 (g/100 g)

Starch Rice flour 61.64 49.32 49.32 49.32 49.32 49.32 49.32
Potato starch 13.68 10.95 10.95 10.95 10.95 10.95 10.95
Cassava starch 20.56 16.43 16.43 16.43 16.43 16.43 16.43
Sour tapioca starch 4.12 3.30 3.30 3.30 3.30 3.30 3.30
Amaranth whole flour 0 20 0 20 0 20 0
Quinoa whole flour 0 0 20 0 20 0 20
Amount of starch 100 100 100 100 100 100 100
% Base flour Demerara sugar 2.73 0 0 0 0 0 0
Sucralose 0 0.003 0.003 0 0 0.002 0.002
Stevia 0 0 0 0.01 0.01 0 0
Acesulfame-K 0 0 0 0 0 0.005 0.005
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 gum 0.35 0.35 0.35 0.35 0.35 0.35 0.35
Pasteurized liquid egg 37.67 37.67 37.67 37.67 37.67 37.67 37.67
Canola oil 10.28 10.28 10.28 10.28 10.28 10.28 10.28
Water at 4  C 82.25 82.25 82.25 82.25 82.25 82.25 82.25

Note: F1: control; F2: amaranth sucralose; F3: quinoa sucralose; F4: amaranth stevia; F5: quinoa stevia; F6: amaranth sucralose/acesulfame-K; F7: quinoa sucralose/ace-
sulfame-K.
 N.M. Machado Alencar et al. / LWT - Food Science and Technology 62 (2015) 1011e1018
ether extract were determined using the AACC (2000) methodol-
ogies. The carbohydrates were calculated by difference.
2.2.4.1. Gross energy. The gross energy (kJ/g) of the samples was
obtained using a Parr calorimetric pump and convert in kcal/g.
2.2.4.2. Analysis of the bread crumb images. Digital images of 1 cm
thick slices of gluten-free bread samples were captured in the jpeg
format. The colored images were first converted to the gray scale
using the program Photo Filter, and then analyzed by the software
Image J. The values for the digitalized images were obtained in
pixels, converted into centimeters using bars of known length (355
pixels equivalent to 1 cm), and the mean area of the alveolus and
the number of alveoli per cm2 was then determined.
2.2.4.3. Statistical analyses. The results of the physicochemical
analyses were evaluated using univariate statistical analysis
(ANOVA), and the means compared by Tukey's test at 5% of sig-
nificance. The calculations were performed using the Statistical
Software Analysis System e SAS (SAS Institute, 2009).
2.3. Time-intensity analysis
Time-intensity analyses were carried out in individual air-
conditioned (22  C) booths with white light. Water was provided
for palate cleansing. Sessions were held at the Laboratory of Sen-
sory Science and Consumer Study of the Food and Nutrition
Department (School of Food Engineering/University of Campinas)
and samples (a half of a slice of bread including crumb and crust)
were presented in white disposable plates with 3-digit numbers
randomly coded.
The time-intensity analysis was performed for the sweet taste
and bitter taste stimuli of the gluten-free bread samples. The two
attributes were evaluated in separate instances.
Approval for the study was obtained from the Ethics Committee
of the University of Campinas, and all volunteers gave a written
consent.
2.3.1. Selection and training of assessors
A preselection of candidates was carried out using Wald's
sequential analysis (Meilgaard, Civille, & Carr, 2004). Two com-
mercial gluten-free bread samples were used and previously tested
to obtain a 1% of significant difference level. Triangle difference
tests were applied with 28 consumers using these gluten-free
bread samples (Gomes, Pflanzer, Felício, & Bolini, 2014). After the
preselection, fourteen assessors were chosen.
All the assessors determined the references by a consensus, and
they were then trained with respect to the product attributes using
identified references (Table 2). The direct contact of the individuals
with the reference of maximum intensity for each stimulus (sweet
taste and bitter taste) led to training for the formation of sensory
memory and equalization among assessors. The panel was trained
in six 1 h training sessions, in which each assessor was trained with
respect to the program and movement of the mouse in the analyses.
2.3.2. Selection of the assessor and evaluation of attributes by time-
intensity analysis
In the selection of the assessors, the analysis of variance
(ANOVA) was applied for each panelist and each parameter in each
stimulus separately, and 12 assessors of 14 were selected to
participate based on their discriminative power, repeatability and
agreement with the team Dama
sio and Costell (1991), verified by
their discriminating capability (p < 0.50) and repeatability
(p > 0.05), while an individual consensus was also considered. For
this selection, the assessors evaluated the attributes by way of a
1013
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 None sweetness: Biscuit with fibers
of sucrose solution. Triunfo® (SP, Brazil), less salt e water type.
A lot sweetness: Toast lightly sweet
Bauduco® (SP, Brazil).
Bitter taste Characteristic taste None bitterness: Biscuit with fibers
of caffeine solution. Triunfo® (SP, Brazil), less salt e water type.
Very bitterness: Bread without gluten with
40 g/100 g 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, 30 ml 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 min and baked in electric
oven during 20 min at 220  C temperature.
monadic presentation with four repetitions, registering the in-
tensity of the attribute on a structured linear scale from zero to nine
(0 ¼ none, 4.5 ¼ moderate, and 9 ¼ strong) on the computer screen,
as a function of the time lapsed.
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 the second signal, the assessor swallowed the sample and
the third signal indicated the end of the test. The time parameters
used in the stimulus evaluation steps, registered on a 9 cm scale
were: 10 s of initial waiting, 30 s of residence in the mouth and 60 s
after swallowing.
The 12 assessors evaluated the samples by way of a balanced
complete block design (Macfie, 1989), in a monadic way with three
repetitions. Data collection for the time-intensity analysis were
carried out on a computer using the software Time Intensity
Analysis of Food and Tastes (TIAF) (Universidade Estadual de
Campinas, 2012).
The curve data obtained were keyed in on the spreadsheet of the
Excel for Windows program and analyzed by the SAS statistical
program (SAS Institute, 2009). The curve parameters evaluated
were: Imax e maximum stimulus intensity recorded by the
assessor; Tmax e time when the maximum intensity was recorded
by the assessor; Area e total area of the time curve  intensity; Ttot
e total duration time of the stimulus. The parameters were evalu-
ated using the analysis of variance (ANOVA) and Tukey's means test.
3. Results and discussion
3.1. Physicochemical analyses
Tables 3 and 4 show the mean values obtained in the physico-
chemical analyses and image analyses of the gluten-free bread
samples.
3.1.1. Specific volume
The loaves weighed approximately 220 g and showed a specific
volume varying from 2.30 to 2.88 cm3/g, as described in Table 3. Low
values were observed for this parameter. The sample containing
quinoa flour and the sweetener stevia presented the highest mean
for specific volume, but was not statistically different (p < 0.05) from
the control, amaranth and sucralose, amaranth and sucralose-
acesulfameeK, quinoa and sucralose-acesulfame-K loaves.
1014 N.M. Machado Alencar et al. / LWT - Food Science and Technology 62 (2015) 1011e1018

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 (g) L* a* b* Water activity Gross energy

(cm3/g) kcal/100 g

Control 2.55abc ± 0.02 470.89abc ± 76.86 79.00a ± 1.54 0.29c ± 0.11 19.16b ± 0.35 0.91ab ± 0.00 243.12c ± 2.31
Amaranth sucralose 2.58abc ± 0.07 396.01c ± 13.60 76.98ab ± 1.78 1.08b ± 0.42 21.91b ± 1.78 0.91ab ± 0.00 248.19bc ± 1.95
Quinoa sucralose 2.36bc ± 0.06 568.97a ± 85.15 76.51abc ± 1.28 0.47bc ± 0.20 21.91ab ± 1.01 0.92a ± 0.00 231.53d ± 1.81
Amaranth stevia 2.30c ± 0.07 394.76c ± 7.33 75.25bcd ± 1.33 1.12b ± 0.11 21.65ab ± 1.29 0.91ab ± 0.01 248.85bc ± 1.06
Quinoa stevia 2.88a ± 0.29 259.33d ± 11.53 73.43cd ± 1.47 0.63bc ± 0.27 22.38a ± 0.95 0.91ab ± 0.00 261.91a ± 2.02
Amaranth sucralose-acesulfame-K 2.65abc ± 0.09 546.36ab ± 67.89 74.13cd ± 0.80 1.91a ± 0.42 24.20a ± 0.79 0.91ab ± 0.01 254.17b ± 0.01
Quinoa sucralose-acesulfame-K 2.67ab ± 0.03 447.89bc ± 43.83 72.05d ± 1.23 1.18ab ± 0.36 24.37a ± 1.31 0.90b ± 0.01 225.98d ± 1.80

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

With the exception of the loaves containing amaranth flour and
the sweetener stevia, the specific volumes of the other samples did
not vary significantly (p < 0.05) from the value obtained for the
control. A similar result was found by Sciarini, Ribotta, Leo

n, and
Pe
rez (2012), who tested different hydrocolloids in gluten-free
bread, and found no statistical difference between the specific
volume of the loaves made using xanthan gum and the control
loaves (with no addition of hydrocolloid). This parameter is
important for acceptance by consumers, because loaves with higher
specific volume are usually more preferred.
3.1.2. Firmness
The gluten-free breads evaluated in this study had a starch-rich
base formulation, and the alterations in the starch granules during
baking determined the crumb structure. Thus, heating causes an
extension of the amylopectin crystals, resulting in swelling of the
granules and alterations in the textural characteristics (Patel,
Waniska, & Seetharaman, 2005). In addition, egg is considered to
be an important ingredient in gluten-free bread formulations, since
it provides softer crumbs when compared to those formulated
without egg, the latter being firmer and less springy (Milde,
Ramallo, & Puppo, 2012). The results for firmness varied from
259.33 to 568.97 g. It can be seen in Table 3 that the sample with
quinoa and stevia was characterized by presenting a lower firmness,
differing statistically (p < 0.05) from the other samples. However,
the sample with quinoa and sucralose presented the higher mean of
firmness and did not differ statistically (p > 0.05) from the control
and amaranth and sucralose-acesulfame-K samples. According to
Esteller, Amaral, and Lannes (2004), the hardness or firmness of the
bread is related to the applied force that causes rupture or defor-
mation of the samples, and is correlated with the human bite. Hager
and Arendt (2013) showed that xanthan gum strengthened the
crumb structure of gluten-free bread, that is, bread supplemented
with xanthan gum showed an increase in hardness.
ingredients making up the formulation and by the baking condi-
tions (Silva, Paucar-Menacho, Vicente, Salles, & Steel, 2009).
The instrumental color analysis as described in Table 3 showed
that the samples presented means varying from 79.00 to 72.05 for
the parameter L*, and were therefore characterized by a lighter
color. This result was expected since gluten-free bread formula-
tions were produced from rice flour and other starches and light
quinoa and amaranth flours. Matos and Rosell (2013) found high
values for L*, varying from 62.24 to 81.50 in gluten-free bread
based on rice flour. All the loaves showed low mean values for
parameter a* and hence presented little reddish coloration. The
samples containing the sweeteners stevia and sucralose-
acesulfame-K and those containing quinoa flour and sucralose
were characterized by higher mean values for parameter b*, sta-
tistically different (p < 0.05) from the control loaves and from those
containing amaranth flour and sucralose. Thus, these samples were
more intensely yellow. The mean values found for the parameters
a* and b* in the present study are similar to those found for other
gluten-free breads containing pseudocereals (buckwheat,
amaranth, quinoa) published in the literature, which were char-
acterized by yellowing of the crumb as compared to the
control bread (Alvarez-Jubete, Auty, Arendt, & Gallagher, 2010;


Wronkowska, Haros, & Soral-Smietana, 2013).
3.1.4. Water activity
According to the results presented in Table 3, the water activity
varied from 0.90 to 0.92 in the gluten-free bread samples evaluated.
The sample produced with the quinoa flour and the sweetener
sucralose stood out for having the lowest mean value (0.90 ± 0.01)
being statistically different (p < 0.05) from the gluten-free bread
sample with quinoa and sucralose. Mariotti, Pagani, and Lucisano
(2013) found values of about 0.99e0.98 for the water activity of
gluten-free bread containing 40 g/100 g of buckwheat flour in its
composition.

3.1.3. Color 3.1.5. Gross energy

The color of bread is considered to be a very important factor in The majority of the energy in bread comes from the starches.
product commercialization, and is directly influenced by the One hundred grams of white bread provide 219 kcal, whilst the

Table 4
Proximate composition averages and averages of image analyses of gluten- and sugar-free bread samples.

Sample Moisture (%) Ash (%) Lipid (%) Protein (%) Carbohydrate (%) Number of alveoli/cm2 Total area alveoli
mm2/cm2

Control 43.94ab ± 2.03 1.26c ± 1.27 6.55d ± 0.12 3.44c ± 0.13 44.81 67.79a 1.99e
Amaranth sucralose 41.08ab ± 1.27 1.65a ± 1.65 7.68bc ± 0.11 4.14ab ± 0.08 45.45 57.35acd 2.66bcde
Quinoa sucralose 45.04ab ± 3.94 1.45b ± 1.45 7.01cd ± 0.04 3.70bc ± 0.06 42.80 48.03ce 3.35ab
Amaranth stevia 41.51ab ± 4.30 1.61ab ± 1.60 7.98ab ± 0.41 4.34a ± 0.24 44.56 56.86abc 2.79bcde
Quinoa stevia 44.94a ± 4.69 1.47b ± 1.47 6.37d ± 0.08 3.70bc ± 0.09 43.52 42.54e 4.25a
Amaranth sucralose-acesulfame-K 43.91a ± 2.03 1.53ab ± 1.53 7.11bcd ± 0.43 3.97abc ± 0.18 43.48 61.02ab 2.13bcde
Quinoa sucralose-acesulfame-K 35.38b ± 1.75 1.67ab ± 1.67 8.69a ± 0.56 4.37a ± 0.34 49.89 50.50bde 3.30ab

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 Tukey's test (p < 0.05).
 N.M. Machado Alencar et al. / LWT - Food Science and Technology 62 (2015) 1011e1018 1015

same weight of whole wheat bread provides 217 kcal (O'Connor,
2012). The caloric values of the breads formulated in the present
study are presented in Table 3, and varied from 231.43 to
261.91 kcal considering 100 g of gluten-free bread samples. The
bread samples formulated with quinoa flour and the sweeteners
sucralose-acesulfame-K presented the lowest caloric indices, and
differed significantly (p < 0.05) from the other samples.
3.1.6. Proximate composition
Table 4 shows that gluten-free breads formulated with pseu-
docereals flours and sweeteners presented higher ash contents,
differing statistically (p < 0.05) from the control sample. In relation
to the lipid contents, the gluten-free bread with quinoa and
sucralose-acesulfame-K presented the highest mean value, which
did not differ statistically (p > 0.05) from the bread with amaranth
and stevia. The control bread presented the lowest mean value for
this parameter, and did not differ significantly (p > 0.05) from the
breads with quinoa and sucralose, quinoa and stevia, and amaranth
and sucralose-acesulfame-K. According to Sanz-Penella,
Wronkowska, Soral-Smietana, and Haros (2013), the incorpora-
tion of 10 g/100 g or more amaranth flour into bread formulations
significantly increases the protein, lipid and ash contents and de-
creases the starch content. The highest protein contents were found
in all the samples produced with amaranth flour and in the bread
formulated with quinoa flour and the sweetener sucralose-
acesulfame-K. Recently, Segura and Rosell (2011) presented the
nutritional composition of 11 types of gluten-free breads, which
contained 0.91e1.05% protein, 2.00e26.10% lipids, 1.10e5.43%
minerals and 68.42e92.96 carbohydrates, showing a similar
nutritional profile to that found in the current study. It can be seen
that the bread samples presented 42.8e49.89% of carbohydrate,
since the gluten-free breads are based mainly on carbohydrates. In
general, the principal components in the formulations are flours
and starches (Matos & Rosell, 2012).

Fig. 1. Digital images of gluten- and sugar-free bread crumbs. A e Control; B e Amaranth and sucralose; C e Quinoa and sucralose; D e Amaranth and stevia; E Quinoa and stevia,
F e Amaranth and sucralose/acesulfameeK; G e Quinoa and sucralose/acesulfameeK. Expansion of 20 for all images.
1016 N.M. Machado Alencar et al. / LWT - Food Science and Technology 62 (2015) 1011e1018

3.1.7. Image analyses

An analysis of the images of the crumb structures of gluten-free
bread samples (Fig. 1 and Table 4) indicates that the formulations
with amaranth and quinoa flours presented a smaller number of
alveoli but with larger areas, being opposite to the control gluten-
free bread. These results also can be seen in Table 4, in which the
number of alveoli presented the highest mean for the control
sample, while the lowest mean value was found for this sample in
relation to the total alveolar area.
The use of buckwheat, amaranth and quinoa flours provides a
structure with larger alveoli in the bread as compared to gluten-
free bread with a starch based formulation (Alvarez-Jubete,
Arendt, et al., 2010). A great variety in structure can be found
amongst the gluten-free breads, some presenting a larger alveolar
area and others a larger number of alveoli. This fact is due to the
diversity of ingredients added to the dough (Matos & Rosell, 2013).

3.2. Time-intensity analysis

3.2.1. Time-intensity analysis for sweet taste stimulus of gluten-free
breads
Table 5 shows the mean values for sweetness obtained for the
gluten-free bread samples in relation to each parameter of the
curve. It was possible to observe that no significant difference
(p > 0.05) was found among samples in relation to the evaluated
parameters: Imax, Tmax, Ttotal, and Area. The control gluten-free
bread sample, which was produced with demerara sugar, pre-
sented mean values for maximum intensity of sweetness statisti-
cally equal to the samples using sweeteners as substitutes of sugar.
Thus, the replacement of sugar by sweetener agents did not alter
the perception of sweetness intensity for the gluten-free breads.
Fig. 2 represents the profile trend for the sweet stimulus as
registered by the assessors. Time-intensity curves showed that the
seven gluten-free bread samples added with quinoa, amaranth and
sweeteners presented similar temporal profiles. The similarities
between the time-intensity curves obtained for the sweeteners
sucralose, stevia and sucralose-acesulfame-K showed their poten-
tial to substitute sucrose. Morais, Cruz, and Bolini (2013) evaluated
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
Fig. 2. Time-intensity curves and characteristics of sweetness stimulus for gluten and
sugar free bread samples.
the time-intensity profile of gluten-free breads in which the sugar
was substituted by prebiotics and sweeteners. It was shown that
the assessors perceived a more intense sweet taste in the breads
containing demerara sugar, followed by those sweetened with
fructooligosaccharides, which showed a similar behavior for the
perception of sweetness throughout the test.
3.2.2. Time-intensity analysis for bitter taste stimulus of gluten-free
breads
Table 5 shows the mean values for bitterness obtained for the
gluten-free bread samples in relation to each parameter of the
time-intensity curve registered for this stimulus, and Fig. 3 repre-
sents the profile trend for the bitter stimulus as registered by the
assessors. No significant difference (p > 0.05) was observed be-
tween the samples for the maximum intensity time (Tmax). A
significant difference (p < 0.05) was observed among samples in
relation to Imax. The sample developed with quinoa and sucralose-
acesulfame-K presented a higher mean value for intensity of
bitterness followed by the samples with quinoa and sucralose and
quinoa and stevia. This result may relate to the addition of quinoa
Area

Sweetness
Control 5.01a 25.91a 43.11a 136.35a
Amaranth stevia 4.92a 28.25a 45.06a 141.66a
Quinoa stevia 5.15a 28.55a 45.98a 148.51a
Amaranth sucralose 4.60a 27.14a 42.78a 122.67a
Quinoa sucralose 4.84a 26.09a 46.37a 142.97a
Amaranth sucralose acesulfame-K 4.81a 27.68a 44.38a 131.99a
Quinoa sucralose-acesulfame-K 4.85a 27.26a 46.37a 131.75a
MDS 0.84 4.28 5.51 32.07a
Bitter
Control 3.245d 25.92a 37.67b 86.19c
Amaranth stevia 4.37cd 26.65a 39.85ab 115.91abc
Quinoa stevia 4.32bc 25.68a 41.39ab 115.13abc
Amaranth sucralose 3.92cd 27.41a 39.58ab 102.16bc
Quinoa sucralose 5.03ab 26.94a 41.01ab 134.18ab
Amaranth sucralose acesulfame-K 4.06cd 25.96a 39.06ab 102.48bc
Quinoa sucralose-acesulfame-K 5.40a 28.32a 45.23a 147.50a
MDS 0.96 4.98 6.46 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; Ttotal: total duration time of
the stimulus; Area: area of the time curve  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% Fig. 3. Time-intensity curves and characteristics of bitter stimulus for gluten and sugar
level. MDS: minimum significant difference in Tukey's test p < 0.05. free bread samples.
 N.M. Machado Alencar et al. / LWT - Food Science and Technology 62 (2015) 1011e1018 1017

flour with perceived increasing on bitterness intensity. On the other
hand, the control sample presented a lower mean value for in-
tensity of bitterness, as can be seen in Fig. 3, and did not differ
statistically (p > 0.05) from the samples with amaranth and
sweeteners. These data may be observed using the time-intensity
curves, which show how similar their profiles are in the behavior
of bitterness perception.
3.2.3. Bitterness and sweetness temporal profile in gluten and
sucrose-free breads
An analysis comparing the time-intensity profile of each sample
in relation to bitterness and sweetness allowed evaluating the
samples individually in relation to the stimuli simultaneously, on
the same graph. It can be seen, in Fig. 4, that in exception of the
breads containing quinoa, sucralose and sucralose-acesulfame-K,

Fig. 4. Time-intensity profile of gluten and sugar free bread samples for sweetness and bitterness stimuli.
1018 N.M. Machado Alencar et al. / LWT - Food Science and Technology 62 (2015) 1011e1018
the sweet stimulus was perceived with greater intensity by the
assessors. However, the use of the sweeteners showed a greater
duration of the stimulus, suggesting a residual sweetness for these
samples. Differently, the bread samples containing quinoa, sucra-
lose and sucralose-acesulfame-K were perceived as having a
greater bitter taste.
Fig. 4 shows that the samples which presented the more similar
temporal profiles in relation to bitterness and sweetness when
compared with the control sample were the samples with
amaranth and the sweeteners. These results show that for gluten-
free breads produced with sugar replacement by sweeteners and
partial replacement of starch by pseudocereals, the addition of
amaranth in formulations with sweeteners led to a more similar
temporal sensory profile in relation to a conventional one (with
demerara sugar and starch based), than the addition of quinoa.
4. Conclusions
The bread samples containing pseudocereals and sweeteners
presented values for specific volume, firmness and water activity
similar to those of the control formulation. It can be seen that the
bread samples containing amaranth and quinoa presented greater
amounts of proteins, lipids and ash, improving their nutritional
profile.
The bread samples developed with demerara sugar showed a
similar behavior with respect to the perception of sweetness to
those developed with sweeteners. With respect to the perception of
bitter taste, the bread samples containing quinoa and the sweet-
eners sucralose, sucralose-acesulfame-K and stevia presented a
greater intensity for this stimulus. However, in the analysis of the
multiple time-intensity 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 gluten-free breads. This research
opens up new opportunities for the gluten- and sucrose-free bak-
ery industry, showing possibilities in developing gluten-free breads
for a group of the population with special needs, as celiacs, gluten
intolerants, diabetics, and/or both of them.
Acknowledgments
The present study was carried out with the support of the
Brazilian National Research Council (CNPq).
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