Journal of Food Engineering 111 (2012) 590–597

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Journal of Food Engineering

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Incorporation of several additives into gluten free breads: Effect on dough

properties and bread quality
L.S. Sciarini, P.D. Ribotta, A.E. León, G.T. Pérez ⇑
CONICET, Facultad de Ciencias Agropecuarias, Universidad Nacional de Córdoba, 5000 Córdoba, Argentina

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 work was to assess the effect of emulsifiers, hydrocolloids and enzymes on gluten-
Received 29 August 2011 free dough rheology and thermal properties and bread quality, while relating dough properties parame-
Received in revised form 24 February 2012 ters to bread technological quality. Breads were based on rice flour, cassava starch and full-fat active soy
Accepted 6 March 2012
flour, with 65% or 75% (flour-starch basis) of water incorporation. Additives used were emulsifiers
Available online 21 March 2012
(diacetyl tartaric acid ester of monoglycerides – DATEM and sodium stearoyl lactylate – SSL), enzymes
(glucose oxidase and a-amylase) and hydrocolloids (xanthan gum, carboxymethylcellulose, alginate
Keywords:
and carrageenan). Results showed that additive incorporation modified dough behavior, evidenced by
Emulsifiers
Enzymes
different calorimetric and rheological properties. Besides, the electrophoretic pattern of dough extracted
Hydrocolloids proteins changed with glucose oxidase addition. These modifications resulted in breads with different
Gluten free dough characteristics, such as specific volume, firmness and firming rate, and crumb structure. Nonetheless,
Gluten free bread they did not necessarily show better quality parameters than the control bread. The control dough dis-
played good performance for obtaining gluten-free breads of acceptable volume, crumb structure and,
principally, with lower hardening rate during storage. Contrary to widespread opinion, this work shows
that the presence of additives is not essential for gluten-free bread production. This fact provides new
perspectives to the gluten free market at the moment of selecting raw materials and technological
parameters, reducing production costs and facilitating gluten free products development.
Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction (Demirkesen et al., 2010). They are composed of hydrophilic and

hydrophobic residues which allow the interaction of two chemi-
Celiac people are unable to consume certain gluten proteins cally different phases. Thus, surface tension between two immisci-
from cereals – such as wheat, rye, barley, kamut, spelt – and hy- ble phases is reduced by emulsifier presence allowing the formation
brids like triticale. The most common cereal flours used for gluten of an emulsion (Krog, 1981; Flack, 1987; Dziezak, 1988). When
free bread production are rice (Gujral and Rosell, 2004a,b; Marco emulsifiers are used in breadmaking, they contribute to increase
and Rosell, 2008; Renzetti and Arendt, 2009), sorghum (Schober the stability of a thermodynamically unstable system (Gómez
et al., 2005) and corn flours (Renzetti et al., 2008). Andean crops et al., 2004). Some emulsifiers have already been incorporated into
(Torbica et al., 2010) and tubers such as potato and cassava gluten free formulations. Onyango et al. (2009) made gluten free
(Sánchez et al., 2002; Ballesteros López et al., 2004; Ribotta et al., bread based on pregelatinized cassava starch and found that emul-
2004) have also been used. In general, breads formulated with sifier addition reinforced dough structure and decreased crumb
gluten free raw materials include high water incorporation; in firmness as well. This behavior was also reported by Demirkesen
the literature water addition ranges between 65% (Ribotta et al., et al. (2010). Enzymes are currently being added to gluten-free
2004) and 110% (Marco and Rosell, 2008). systems as a means of modifying protein functionality (Gujral and
Nevertheless, the absence of gluten produces technological Rosell, 2004a,b; Moore et al., 2006; Renzetti et al., 2008), as the for-
problems in the production of baked goods. To counteract these mation of a continuous protein network is considered a key factor
technological problems, several additives have been employed to in enhancing gluten free flours performance for breadmaking. Thus,
mimic gluten properties. Emulsifiers are used in the baking indus- glucose oxidase incorporation has been studied as a polymerizing
try because of their ability to interact with different flour compo- agent with varying results, depending on the raw material
nents and other dough ingredients, resulting in softer crumbs employed (Gujral and Rosell, 2004b; Renzetti and Arendt, 2009).
The a-amylase has been extensively used to delay amylopectin ret-
rogradation. Different hydrocolloids have been added to leavened
⇑ Corresponding author. Tel.: +54 351 4334116; fax: +54 351 4334118. gluten free products with positive results on crumb structure, taste,
E-mail address: gaperez@agro.unc.edu.ar (G.T. Pérez). global acceptability and shelf life. It has been reported that

0260-8774/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jfoodeng.2012.03.011
 L.S. Sciarini et al. / Journal of Food Engineering 111 (2012) 590–597 591

hydrocolloids improve dough development and gas retention increase air incorporation into the dough); dough was weighed
through the increase in system viscosity, producing loaves with into aluminum cups (60 g) and proofed again under the same con-
higher specific volume (Lazaridou et al., 2007; Marco and Rosell, ditions (30 min, 30 °C, and 85% relative humidity). Finally, they
2008; Peressini et al., 2011). However, it is worth highlighting that were put into a rotational oven (Ciclo Ingeniería, Argentina) and
the effect of different additives is highly dependent on the raw baked at 180 °C for 30 min. Once baked, breads cooled for 2 h (until
material used, the nature and quantity of additive used and water room temperature was reached). Breadmaking was performed in
availability, being very difficult to predict the real effect of each duplicate.
additive on different formulations. Thus, the objective of this work
was to assess the effect of emulsifiers, hydrocolloids and enzymes 2.3. Dough properties
on gluten-free dough rheology and thermal properties and bread
quality, while relating dough properties parameters to bread tech- 2.3.1. Large deformation rheology: resistance to penetration
nological quality. The force required to penetrate the dough was determined using
a TA-XT2i texturometer (Stable Micro Systems, United Kingdom)
2. Materials and methods equipped with a 25 kg cell. Samples were prepared as for bread-
making, and 40 g of the resultant dough were weighed into plastic
2.1. Materials flasks and proofed (60 min, 30 °C, 85% relative humidity). To deter-
mine penetration force, fermented dough was compressed until the
Gluten free breads were formulated with rice flour (Nora’s probe (35 mm diameter) disrupted the dough surface structure, pe-
Skills, Argentina; 8.11% proteins, 0.23% ash, 0.28% crude fiber, netrating into the sample, at 5 mm/s. Fig. 1 shows a representative
0.80% lipids, 79.63% carbohydrates, 10.95% moisture), cassava penetration plot. In the first part of the curve, probe is considered to
starch (Señor de Sipan, Argentina; 0.24% proteins, 0.09% ash, compress the dough without disrupting its structure, up to the
0.21% crude fiber, 0.01% lipids, 86.59% carbohydrates, 12.87% mois- point where a threshold force is achieved, and dough resistance
ture) and full-fat active soy flour (NICCO, Argentina; 36.41% to penetration is broken. To obtain this threshold value, two linear
proteins, 4.72% ash, 2.83% crude fiber, 19.80% lipids, 30.26% carbo- regressions were carried out in each of the two parts of the curve;
hydrates, 5.98% moisture); compressed yeast (Dánica, Argentina), these regressions represented the ideal behavior of the dough.
shortening (Dánica, Argentina) and salt (Dos Anclas, Argentina). The intersection of both straight lines was considered as dough
The additives employed were: emulsifiers: sodium stearoil-2- resistance to penetration under ideal conditions. Dough prepara-
lactilate (SSL) and diacetyl tartaric acid ester of monoglyceride tion was performed in duplicate, and three determinations were
(DATEM) were obtained from Alpha emulsionantes (Argentina). performed in each dough batch.
Enzymes: glucose oxidase (GOX) and a-amylase (Am) were pur-
chased from Novozyme (Denmark). Hydrocolloids and emulsifiers 2.3.2. Small deformation rheology: frequency sweep
were of food grade, and enzymes were of analytical grade. Hydrocol- Rheometric experiments were performed with an oscillatory
loids: xanthan gum (X), carboxymethylcellulose (CMC), carrageenan rheometer (Anton Paar, Germany). Frequency sweeps were carried
(C) and alginate (Al). X, C and Al were provided by Saporiti S.A. out at 0.1–10 Hz, 0.05% strain and 30 °C (viscoelastic linear range
(Argentina), and CMC was obtained from Latinoquímica Amtex was determined with a previous strain sweep from 0.1% to 100%,
S.A. (Argentina). at a constant frequency of 1 Hz). Plate-plate geometry (25 mm
diameter) was used, with 2 mm gap. Samples were prepared as
2.2. Breadmaking for breadmaking, but without yeast addition. Dough was allowed
to rest for 15 min and then put between plates, and sample excess
Basic bread formulation consisted in 45 g of rice flour, 45 g of was carefully trimmed. To avoid water loss during the determina-
cassava starch, 10 g of soy flour, 2 g of salt, 2 g of shortening, 3 g tion, the exposed edges of dough were covered with vaseline. Be-
of compressed yeast and 65 g of water (except in breads with fore starting the assay, samples were rested for 5 min to allow
hydrocolloid addition, where 75 g of water were used). The level residual stresses relaxation. Dough preparation was performed in
of additive incorporation was selected according to preliminary re- triplicate.
sults (Table 1). Ingredients were put together and mixed in a plan-
etary mixer (Arno, Brazil) for 1 min at 156 rpm and 2 min at 2.3.3. Differential scanning calorimetry (DSC)
214 rpm. The dough obtained was proofed for 30 min (30 °C and For starch gelatinization studies, dough was prepared as for
85% relative humidity). After this process, dough was mixed again breadmaking but without shortening addition; then, it was proofed
for 1 min at 156 rpm (this second mixing is carried out to redistrib- (60 min, 30 °C, 85% relative humidity). Approximately 30 mg of
ute air cells and nutrients to improve yeast’s activity, and to sample were weighed into aluminum pans and hermetically

Table 1
Additives employed in gluten-free bread formulations.

Group Additive Code g/100 g flour-starch

Emulsifiers Diacetyl tartaric acid ester of monoglyceride DATEM 1
Sodium stearoil-2-lactilate SSL
Enzymes Glucose oxidase GOX 1 0.003
GOX 2 0.03
a-Amylase Am 1 0.0006
Am 1 0.001
Hydrocolloids Xanthan gum X 0.5
Carboxymethylcellulose CMC
Carrageenan C
Alginate Al
592 L.S. Sciarini et al. / Journal of Food Engineering 111 (2012) 590–597

measured points in a force-time plot. Four slices from two different

bread loaves were analyzed in each breadmaking batch.

2.4.3. Crumb structure

Digital images from breads were obtained from slices of 15 mm
thick using a scanner (HP Scanjet G3010, Palo Alto, USA), with
600 dpi resolution. Images were analyzed using ImageJ Software
1.41o (National Institutes of Health, USA). Image binarization was
carried out according to Ribotta et al. (2010). Cell average area
(mm2) and the number of cell/mm2 were determined. The ratio of
small cells (0.15 < x < 2.00 mm2) to large cells (2.00 < x < 10.00
mm2) was calculated and it was used as a measure of crumb unifor-
mity. Six slices from three different bread loaves were analyzed in
each breadmaking batch.

2.5. Statistical analysis

A completely randomized design was used, with a classifying

and a response variable (in this design, it is assumed that the error
is normally distributed with a mean = 0, and constant variance).
Mean values ± standard deviation are presented. The data obtained
were statistically treated by analysis of variance (ANOVA) and the
Fig. 1. Representative penetration test plot. The intersection of the two straight- means were compared by the Fisher LSD test at a significance level
lines was considered as the force required for the probe to penetrate the dough.
of 0.05 (coefficient of variation due to sample preparation was low-
er than 10%). A correlation test was made to evaluate the relation-
ship between variables (p < 0.05). These tests were carried out with
sealed. Pans were then heated in the DSC using a temperature pro-
INFOSTAT statistical software (2011).
file similar to that measured in the crumb core during baking (León
et al., 1997). Temperature profile was as follows: 2 min at 30 °C for
sample stabilization, heating from 30 to 110 °C at a heating rate of 3. Results and discussion
11.7 °C/min, and 5 min at 110 °C. Starch gelatinization parameters
(To, peak width and DH) were obtained from the transition endo- 3.1. Dough analyses
therm. To analyze amylopectin retrogradation, pans were stored at
4 °C for 7 days. After this period, pans were reheated in the DSC 3.1.1. Rheological studies
from 30 to 120 °C at a heating rate of 5 °C/min. To, peak width Table 2 shows the effect of different additives on dough rheo-
and DH were also obtained. Dough was prepared in duplicate logical properties. As shown, emulsifier incorporation affected
and three pans were prepared in each dough batch. dough behavior during fermentation, though the effect was differ-
ent from one emulsifier to another. Thus, when SSL was used,
2.3.4. Dough protein extraction and separation dough resistance was higher than the control (no additive), while
Dough was prepared as for breadmaking. Protein extraction was the result was just the opposite when DATEM was added. For all
carried out with two different solutions: TRIS/HCl 0.5 M, pH 8.8; gluten-free doughs, G0 was higher than G00 in all the frequency
and TRIS/HCl + 2% SDS. Dough:solvent ratio was 1:10 (w/v). Sam- range studied, which was indicative of a solid-elastic behavior
ples were vortexed for 30 min and then centrifuged 20 min at (Fig. 2). Doughs with both emulsifiers presented higher G0 and G0 0
3000g. The supernatant was mixed with sample buffer (without values than the control. SSL dough had higher G0 than DATEM,
2-mercaptoethanol) and proteins were separated using SDS–PAGE while the latter one had lower tan d. The higher values found in dy-
under non-reducing conditions (Ribotta et al., 2005). namic moduli (G0 and G00 ) of doughs with emulsifiers regarding
control dough, clearly indicate that the presence of emulsifiers
2.4. Bread quality introduces new interactions into the system, and that their effect
will also depend on the type of hydrophilic and hydrophobic inter-
2.4.1. Specific bread volume (SBV) actions established. It has been reported that in wheat based sys-
The volume of each bread loaf was determined by rapeseed dis- tems, emulsifiers facilitate the interaction between lipids,
placement. Specific volume was obtained by dividing bread vol- proteins and starch (Jacobsberg et al., 1976), possibly due to their
ume/bread weight. Three measurements of each breadmaking amphiphilic nature, and that these interactions are responsible
replication were performed. for dough reinforcement. The same phenomenon could take place
in gluten-free systems, were starch and proteins are the main
2.4.2. Crumb firmness dough components.
To assess crumb firmness, breads were longitudinally cut 2 h Enzyme addition significantly reduced dough resistance after
after baking, and two slices of 15 mm thick were obtained from fermentation. Nevertheless, when analyzing frequency sweeps, it
the center of each loaf. Firmness was measured using an Instron is observed that both GOX doses increased dough consistency
Universal Texture Machine (Instron, USA) equipped with a (given by higher G0 and G00 values), and that tan d was significantly
25 mm diameter probe, at a rate of 5 mm/s and 40% compression. reduced. GOX catalyzes the oxidation of glucose to give glucono-
Firmness was defined as the maximum force obtained during com- lactone and H2O2. The H2O2 thus formed oxidizes sulfhydryl groups
pression. To obtain hardening rate, firmness measurement was present in proteins, inducing protein cross-linking through the forma-
performed 2, 24 and 72 h after baking. Breads were stored in sealed tion of disulfide bonds. H2O2 is also proposed to be involved in the oxi-
plastic bags at 25 °C. Hardening rate was calculated as the slope of dative gelation of water soluble pentosans (Hoseney and Faubion,
the straight-line obtained from the regression of the three 1981), which induces the formation of a protein/polysaccharide
 L.S. Sciarini et al. / Journal of Food Engineering 111 (2012) 590–597 593

Table 2
Resistance values after fermentation; elastic (G0 ) and viscous (G0 0 ) moduli and tan d values at 1 Hz for gluten-free doughs.

Additive group Additive Resistance (g) Rheometry

G0 (kPa) G0 0 (kPa) tan d
a
Emulsifiers None 46.3 ± 3.5b 29.8 ± 1.6a 6.8 ± 0.2a 0.230 ± 0.006b
DATEM 28.7 ± 1.8a 84.6 ± 6.9a 15.9 ± 0.5a 0.188 ± 0.009a
SSL 58.8 ± 1.4c 165.5 ± 17.5b 41.1 ± 4.6b 0.246 ± 0.014b
Enzymes None 46.3 ± 3.5c 29.8 ± 1.6bc 6.8 ± 0.2b 0.230 ± 0.006a
GOX 1 27.4 ± 1.23a 38.2 ± 4.3c 7.9 ± 1.1b 0.209 ± 0.008ab
GOX 2 33.9 ± 2.3b 51.1 ± 2.1d 10.3 ± 0.2c 0.201 ± 0.001a
Am 1 27.3 ± 0.4a 20.8 ± 1.3ab 4.5 ± 0.0a 0.217 ± 0.001bc
Am 2 24.7 ± 1.1a 17.4 ± 2.2a 3.9 ± 0.4a 0.223 ± 0.002cd
Hydrocolloids None 46.3 ± 3.5c 29.75 ± 1.62a 6.8 ± 0.2a 0.230 ± 0.006b
X 35.6 ± 4.3b 30.57 ± 4.24a 7.5 ± 2.7a 0.243 ± 0.003bc
CMC 28.5 ± 1.2a 24.15 ± 3.31a 6.5 ± 0.8a 0.266 ± 0.002d
C 22.9 ± 0.7a 60.82 ± 4.18b 12.9 ± 1.3b 0.209 ± 0.006a
Al 25. 1 ± 1.42a 21.64 ± 2.15a 5.4 ± 0.4a 0.245 ± 0.019c
a
Different letters within a column and within the same additive group are significantly different (p < 0.05).

Doughs were formulated with 65% and 75% (flour/starch basis)

of water addition; doughs evaluated were those used for bread-
making. Thus, considering that dough with hydrocolloids included
higher water amount, it was expected to present lower resistance
to penetration. Among doughs with hydrocolloids, X showed the
highest resistance, followed by CMC, Al and C (p < 0.05). Xanthan
gum is known for its thickening properties; it is accepted that in
aqueous systems it adopts a helix conformation which turns the
molecule rigid, and that this conformation plays an important role
on xanthan solution behavior, including high viscosities (Millane
and Wang, 1990).
Considering frequency sweeps, hydrocolloids, with the excep-
tion of C, did not lead to a significant change in G0 and G00 values
(1 Hz) with respect to the control dough. Lazaridou et al. (2007),
working with rice flour and corn starch based doughs with differ-
ent water amounts (130, 140 and 150 g/100 g solids), found a de-
crease in elastic modulus as the water amount increased. This
behavior is well documented in wheat systems (Phan-Thien and
Safari-Ardi, 1998; Autio et al., 2001), where higher water ratios
lead to a diminution in G0 and G00 , without modifying tan d, thus
concluding that water has mainly a plasticizing effect, while dough
structure is unaltered. The C dough had higher G0 values than con-
trol dough; the same trend was found for viscous modulus (G00 );
tan d values were different from one sample to another: they were
higher for CMC, followed by Al, the control and X (p > 0.05), and
lowest for C. Molina Ortiz et al. (2004) studied the behavior of gels
based on soy protein isolate and carrageenan, and found a specific
interaction between proteins and the hydrocolloid which led to the
formation of a more viscoelastic gels.
The correlation between rheology at small and large deforma-
Fig. 2. Dynamic moduli (G0 and G0 0 ) as a function of frequency for all studied tions is still controversial in the literature (Tronsmo et al., 2003;
samples.
Dobraszczyk and Salmanowicz, 2008; Angioloni and Collar,
2009). Different interactions are established between dough com-
ponents. If the molecular interactions that are established are
cross-linked entity, responsible for the increment in the consistency strong enough, they may result intact even at high deformation
of the system. Thus, the increase in G0 and G00 could be due to protein conditions. On the contrary, weak interactions may disrupt under
cross-linking and/or pentosans oxidative gelation. a-Amylase these deformation conditions. Thus, the relationship between both
hydrolyzes a-(1–4) bonds present in starch, producing low molec- types of assays may be considered a function of the interactions
ular weight a-dextrins. Am incorporation led to a reduction in established among dough components. In this work, no correlation
dough resistance during fermentation. G0 and G00 were also reduced, was found.
especially for the highest dose. Damaged starch is a starch fraction
resulting from the milling process, and is susceptible to enzyme 3.1.2. Effect of GOX on protein fraction
hydrolysis, although native starch can, as well, turn into enzyme Fig. 3 shows SDS–PAGE gel under non-reducing conditions from
substrate during gelatinization (Ferrand, 1964). Thus, it was not TRIS/HCl and TRIS/HCl + SDS dough-extracted protein fractions. In
surprising to find lower resistance and G0 values as a consequence GOX samples, a high molecular weight band (arrow) which is
of Am addition/action on susceptible starch fraction. absent in the control sample, possibly due to enzyme action, can
594 L.S. Sciarini et al. / Journal of Food Engineering 111 (2012) 590–597

was observed. It has been previously reported (Sciarini et al., 2012)

that soy proteins diminish cassava starch retrogradation. Thus, the
possible interaction of emulsifiers with proteins and/or starch may
disrupt soy proteins and cassava starch interaction. Retrogradation
peak width was reduced, suggesting the formation of crystallites
with similar stability.
Starch gelatinization was not modified by enzymes addition;
only the presence of Am increased To. Durán et al. (2001) evaluated
the incorporation of maltodextrins with different degrees of poly-
merization (3–7 DP) into different starches; they found an increase
in gelatinization temperature and ascribed this behavior to a stabi-
lizing effect of oligosaccharides on starch amorphous regions, while
they found no effect on DH. Am did not affect the retrogradation
process, as compared to the control sample. Goesaert et al. (2009)
have questioned the impact of a-amylase on starch recrystalliza-
tion, considering that it has little effect on the outer, crystallizable
branches of amylopectin. In this sense, these authors suggested
Fig. 3. Non-reducing SDS–PAGE gel of TRIS/HCl (TRIS) and TRIS/HCl + SDS (SDS)
the use of maltogenic amylase (exoamylase) which degrades amy-
extracted proteins. MWS, molecular weight standard. lopectin chains producing a-maltose almost exclusively. GOX
addition augmented amylopectin retrogradation. Again, the possi-
ble modification of soy proteins by GOX action may negatively
be observed. Gujral and Rosell (2004a) have also informed about affect the interaction between soy proteins and starch.
the modification of protein fraction from rice flour after GOX incor- Hydrocolloid presence did not affect starch gelatinization behav-
poration, obtaining higher molecular weight polymers while they ior; no significant differences were observed in To or DH. Hydrocol-
observed a reduction in free sulfhydryl groups, associated to disul- loids are expected to compete with starch for water uptake (thus
fide bond formation. Besides, sulfhydryl groups are also present in modifying gelatinization process) due to their high hydrophilic nat-
soy proteins (Kinsella, 1979) which may potentially be modified by ure, but since doughs with hydrocolloids were prepared with higher
H2O2. No differences were found in electrophoretic pattern of water amount (75% vs. 65% for the control sample), it was difficult to
protein extracted from doughs with hydrocolloids, emulsifiers or draw a direct comparison. Ferrero et al. (1996) assessed the effect of
a-amylase (data not shown). different hydrocolloids incorporation on corn starch gelatinization
and retrogradation; they found that at low hydrocolloid:starch ra-
3.1.3. Calorimetric behavior tios (1:10), in excess water, gelatinization parameters were not
As shown in Table 3, gelatinization enthalpy was reduced by modified, while higher ratios (1:2 and 1:1) produced a shift toward
emulsifier addition, and this effect was more evident for DATEM higher temperatures, while peak width was also increased. Regard-
which, besides, shifted the gelatinization peak toward higher tem- ing the retrogradation process, DH was increased by hydrocolloid
peratures. Eliasson (1986) reported that the inclusion of emulsifiers addition, indicating higher amylopectin recrystallization during
such as SSL delayed starch gelatinization; they also found a decrease storage. It has been reported that DHret presents a bell-shaped
in DH and attributed this finding to the simultaneous occurrence of response as a function of moisture content, with minimum values
exothermic phenomena, such as the amylose-emulsifier complex at extreme moisture concentrations (higher than 90% and lower
formation. Ghiasi et al. (1982a,b); observed a restriction in wheat than 20%) and a maximum value at approximately 50% of water
starch swelling in the presence of SSL, which partially explains the amount (Zeleznak and Hoseney, 1986). Breadcrumb presents mois-
higher transition temperatures observed. It is widely accepted that ture contents between 35% and 45% (Rogers et al., 1988; He and
the addition of emulsifier (mainly SSL and DATEM) leads to a Hoseney, 1990; Baik and Chinachoti, 2000; Ribotta and Le Bail,
decrease in starch retrogradation (Batres and White, 1986; Krog, 2007); under such conditions, higher water amount leads to higher
1981; Eliasson and Ljunger, 1988; Biliaderis and Tonogai, 1991; amylopectin retrogradation. Thus, the higher water content of
Gudmundsson, 1992). Nevertheless, in this work an increase in DHret

Table 3
DSC parameters of gelatinized and retrograded samples.

Additive Baking Retrogradation

DH (J/g solids) To (°C) Peak width (°C) DH (J/g solids) To (°C) Peak width (°C)
None 7.01 ± 0.72ca 66.23 ± 0.29a 22.10 ± 1.88a 1.71 ± 0.22b 42.88 ± 0.97a 16.58 ± 1.28b
DATEM 4.98 ± 0.39a 67.82 ± 0.53b 22.59 ± 0.67a 2.89 ± 0.42a 44.19 ± 0.68a 12.21 ± 1.14a
SSL 5.86 ± 0.37b 66.46 ± 0.49a 21.79 ± 1.49a 2.09 ± 0.39b 43.25 ± 1.73a 15.59 ± 1.71b
None 7.01 ± 0.72a 66.23 ± 0.29a 22.10 ± 1.88a 1.71 ± 0.22bc 42.88 ± 0.97a 16.58 ± 1.28b
GOX 1 7.27 ± 0.58a 66.43 ± 0.35a 23.09 ± 0.96a 2.13 ± 0.29ab 43.79 ± 0.37b 15.89 ± 0.67b
GOX 2 6.98 ± 0.73a 66.67 ± 0.84ab 22.54 ± 0.74a 2.42 ± 0.24a 44.32 ± 0.41b 14.99 ± 0.42a
Am 1 7.19 ± 0.69a 67.33 ± 0.47c 21.59 ± 0.45a 1.68 ± 0.18c 43.15 ± 0.17a 16.54 ± 0.62b
Am 2 7.49 ± 0.76a 67.23 ± 0.20bc 21.21 ± 0.67a 2.11 ± 0.24abc 44.03 ± 0.14b 14.83 ± 0.49a
None 7.01 ± 0.72a 66.23 ± 0.29a 22.10 ± 1.88c 1.71 ± 0.22c 42.88 ± 0.97a 16.58 ± 1.28b
X 7.16 ± 0.69a 67.07 ± 1.15a 18.94 ± 0.29ab 2.09 ± 0.26b 43.17 ± 0.75a 15.14 ± 0.91a
CMC 7.09 ± 0.62a 66.28 ± 0.54a 19.35 ± 1.09b 2.52 ± 0.22a 43.30 ± 0.36a 14.30 ± 0.45a
C 6.23 ± 0.62a 66.45 ± 0.29a 19.67 ± 0.68a 2.62 ± 0.32a 43.07 ± 0.20a 14.39 ± 0.23a
Al 6.55 ± 0.67a 67.14 ± 0.58a 17.96 ± 0.59a 1.92 ± 0.14bc 43.05 ± 0.38a 15.12 ± 0.57a

DH: enthalpy value; To: onset temperature; PW: peak width.

a
Different letters within a column and within an additive group are significantly different (p < 0.05).
 L.S. Sciarini et al. / Journal of Food Engineering 111 (2012) 590–597 595

a SBV similar to the control bread, although protein polymerization

has been observed. Breads with hydrocolloid addition showed poor
technological parameters – such as very low specific volume, high
firmness and dense crumb structure – when 65% of water was
used. Consequently, different water amounts from 65 to 95 g were
evaluated, obtaining the best result (concerning bread volume,
crumb firmness and structure) when using 75%. For this reason,
breads with hydrocolloid addition were made with 75% of water
incorporation. C addition led to the highest SBV among samples
with hydrocolloids, followed by CMC. Breads with X and Al
addition showed the same SBV than the control bread (p > 0.05).

3.2.2. Crumb firmness

From Table 4 it is observed that crumbs with emulsifier were
harder than the control, and the same trend was observed for firm-
ing rate (related to staling). Considering breads with enzymes,
Fig. 4. Images of gluten-free breads. None: control. Bar: 1 cm.
crumb firmness was reduced by the presence of GOX, in agreement
with Gujral and Rosell (2004a) who also found a diminished crumb
samples with hydrocolloids may explain the higher DHret found in firmness when adding GOX. But the firming rate increased with re-
DSC studies. spect to the control bread. As it was already explained, the possible
disruption of soy protein/starch interaction for GOX action may
negatively affect crumb behavior during storage (Sciarini et al.,
3.2. Bread quality 2012). The presence of Am led to reduced initial crumb firmness
and did not modify the firming rate when compared to the control.
3.2.1. Specific bread volume (SBV) Initial crumb firmness and firming rate were reduced with hydro-
The addition of emulsifiers did not lead to an increase in SBV; in colloids incorporation. This effect would not be related to a de-
fact, SSL addition decreased SBV when compared to the control crease in amylopectin retrogradation (DHret was higher for
bread (Table 4). A negative correlation (r = 0.80, p < 0.05) was samples with hydrocolloids), but more likely to a reduction in
found between dough resistance and SBV. In previous works, moisture loss during storage, which retards staling phenomena
Sciarini et al. (2010a,b) observed an opposite trend in gluten free (Rosell et al., 2007). These results are in agreement with Rogers
systems with high water amount (150%, flour basis), where an in- et al. (1988), who found higher crumb firmness and firming rate
crease in batter/dough resistance led to better quality breads, with in breads with lower moisture contents (between 22% and 37%)
increased SBV; this effect was then ascribed to the higher capacity to and this effect was not associated to an increase in amylopectin
retain the gases formed during fermentation. In this work, a signif- retrogradation, which was lower at lower water contents.
icantly lower water amount was used (65–75%, flour basis). It is
natural then that systems with a higher resistance to certain values 3.2.3. Crumb structure
will have more difficulty to expand during proofing and baking. Fig. 4 shows representative images of the gluten-free breads
The lowest Am dose produced an increase in SBV. This effect is obtained. Table 4 presents crumb structure parameters of all the
mainly due to the hydrolysis of the starch fraction leached as a samples studied. DATEM samples showed a lower cell number.
result of gelatinization during baking, reducing dough resistance Breads with SSL systematically presented a big cell near the
with a positive effect on SBV; and, besides, to the production of surface, this effect being characteristic of systems with a rapid
fermentable sugars. On the other hand, the highest dose did not in- water loss; the vapor thus formed exerts certain pressure on the
crease SBV; it produced a higher reduction in dough resistance as forming crumb, producing the collapse of the structure. These
compared to the lowest dose, and this could lead to a decreased breads had lower air area fractions. GOX crumb presented a higher
gas holding capacity. The presence of GOX produced breads with cell number of small size, while the air area fraction was lower.

Table 4
Gluten-free bread quality parameters.

Additive group Additive SBV (cm3/g) Hardness Crumb structure

Initial Firmness (g) Staling rate(g/day) N° cells/mm2 Cell size (mm2) % Cell area Uniformity
b a
Emulsifiers None 1.98 ± 0.05b 249 ± 39a 208.8 ± 2.3a (0.998) 1.30 ± 0.12b 4.26 ± 0.6a 54.9 ± 2.7b 2.07 ± 0.21b
DATEM 1.99 ± 0.09b 280 ± 11a 361.4 ± 77.9b (0.991) 1.12 ± 0.05a 4.96 ± 0.5a 54.9 ± 1.3b 1.31 ± 0.12a
SSL 1.71 ± 0.00a 833 ± 46b 380.0 ± 24.8b (0.997) 1.04 ± 0.05a 4.48 ± 0.3a 45.9 ± 0.9a 2.18 ± 0.17b
Enzymes None 1.99 ± 0.05a 249 ± 39b 208.8 ± 2.3a (0.998) 1.30 ± 0.12a 4.26 ± 0.6b 54.9 ± 2.7c 2.07 ± 0.21a
GOX 1 2.05 ± 0.04ab 169 ± 11a 329.9 ± 16.3b (0.995) 1.60 ± 0.15b 3.17 ± 0.3a 50.8 ± 2.7ab 2.54 ± 0.22bc
GOX 2 2.01 ± 0.04a 169 ± 19b 304.4 ± 37.7ab (0.999) 1.92 ± 0.17b 3.34 ± 0.5a 49.0 ± 4.9a 2.62 ± 0.20c
Am 1 2.15 ± 0.03b 171 ± 15ab 287.3 ± 32.1ab (0.999) 1.34 ± 0.15a 4.05 ± 0.5b 53.2 ± 2.3bc 2.58 ± 0.28bc
Am 2 2.04 ± 0.04ab 229 ± 22ab 316.2 ± 81.8ab (0.999) 1.27 ± 0.05a 4.15 ± 0.2b 53.7 ± 1.9bc 2.32 ± 0.21ab
Hydrocolloids None 1.98 ± 0.05a 249 ± 39c 208.8 ± 2.3c (0.998) 1.30 ± 0.12c 4.26 ± 0.6c 54.9 ± 2.7b 2.07 ± 0.21c
X 1.86 ± 0.04a 162 ± 9b 172.7 ± 13.6b (0.991) 1.07 ± 0.12b 5.38 ± 0.6d 55.8 ± 1.5b 1.68 ± 0.12ab
CMC 2.14 ± 0.02b 113 ± 7a 136.5 ± 4.6a (0.999) 1.67 ± 0.19d 2.73 ± 0.3a 49.0 ± 1.7a 4.32 ± 0.47d
C 2.38 ± 0.09c 132 ± 1ab 170.9 ± 13.0b (0.996) 0.89 ± 0.08a 8.06 ± 0.7e 58.8 ± 2.0c 1.42 ± 0.06a
Al 1.99 ± 0.02a 141 ± 3ab 160.3 ± 14.3ab (0.994) 1.58 ± 0.14d 3.38 ± 0.3b 51.4 ± 2.8a 2.01 ± 0.19bc
a
Values between parentheses correspond to the determination coefficient of the regression straight-line.
b
Different letter within a column and within an additive group are significantly different (p < 0.05). SBV: specific bread volume.
596 L.S. Sciarini et al. / Journal of Food Engineering 111 (2012) 590–597

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