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ABSTRACT
Soybean proteins are ideally suited to enhance the essential amino acid balance of cereal-based
foods. The aim of this investigation was to assess the functionality of different soybean proteins
in maize tortillas using yield, sensorial and textural shelf-life characteristics as criteria to select
the best supplement. Four different defatted soybean flours (SBF1, SBF2, SBF3, and SBF4) and
one soybean protein concentrate (SBC) were added to increase protein content of dry masa flour
between 25 to 30%. The evaluated soybean ingredients depicted Urease Activity between 0.1 to
2.25, Water Absorption Index of 4.02 to 8.34, Protein Dispersability Index of 23 to 75% and Fat
Absorption Index ranging from 2.5 to 3.1. Moisture, crude protein, crude fat and rollability were
not different among produced tortillas, but maximum force after five days of storage was higher
for SBF1 and lower for SBF 3. Control and SBF1 followed by SBC were the best overall
evaluated supplements according to the most relevant parameters for consumers and producers.
Correlation analyses depicted a negative association among yield-related parameters and Protein
Dispersability Index, Urease Activity and Water Solubility, opposite to relationship for texture-
related properties. The best soybean proteins to be used in maize tortilla supplementation should
have, preferably, reduced Water Solubility, Urease Activity and Protein Dispersability Index.
Page 2 of 35
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M - 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1
5
a p e r h a s b e e n p e e r r e v i e w
e d a n d a c c e p t e d f o r p u b l i c a t i o n b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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INTRODUCTION
Protein malnutrition is still an endemic problem in many developing countries around the globe.
It is known to inhibit growth and have a profound and significant effect on physical
development, susceptibility to infectious diseases and brain maturity. An early malnutrition
reduces both the brain and cerebellum sizes (Chase et al 1967; Culley and Lineberger 1968), the
number of neurons and synapsis (Stylianopoulos et al 2002; Warren and Bedi 1984) and the
neuron myelization process mainly due to the lower synthesis of proteolipids, cerebrosides,
sulphatides and plasmalogen of the white substance. Functionally speaking, this reduction is
relevant because myelinized axons transmit information faster than non-myelinized fibers
(Wiggins 1982). Post-mortem inspection of children that were severely affected by Marasmus
found significantly lower levels of brain cholesterol, phospholipids, RNA and DNA (Rosso et al
1970).
Mexico has one of the highest world per capita consumption of maize ( Zea mays L) and the main
food product from this cereal is tortilla. The lower the socioeconomic status, the greater the
dependence on tortillas. Unfortunately, tortillas are not a perfect food because they lack of two
essential amino acids, lysine and tryptophan, and adequate levels of iron, zinc and vitamins A, D,
E and B12. That is the reason why tortillas are ideally suited for nutritional enrichment and
protein fortification (Amaya-Guerra et al 2004, 2006; Serna-Saldívar et al 1988; Stylianopoulos
et al 2002).
Page 3 of 35
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M - 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1
5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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tonnes. Soybeans are mainly channeled to the oil crushing industries in order to extract the oil
and the defatted soybean meal. The full-fat, defatted, concentrates and isolates contain about 42,
49, 65 and 90% protein, respectively. These products are widely applied in the food industry as
important ingredients due to their highly nutritious and desirable functional properties. However,
in many cases, the application of these types of proteins is limited due to incompatibility between
their solubility and other properties (Wang et al 2008). The enhancement of protein quality and
nutritional value of soybean-fortified maize tortillas was first documented with laboratory
animals in the 1970´s (Bressani et al 1974; Bressani et al 1979; Del Valle and Perez-Villaseñor
1974). These early investigations clearly indicated that it was feasible to produce fortified
tortillas with enhanced protein quality. Amaya-Guerra et al (2004, 2006) determined the
physiological development and brain development of rats fed with soybean fortified tortilla-
based diets for two generations. Growth, reproductive performance, brain development and short
and long term memory of rats fed with soybean fortified tortillas were significantly higher than
counterparts fed with regular tortillas. Chavez and Chavez (2004) conducted a two year blind
study with humans that evaluated the effect of soybean fortification and enrichment with selected
micronutrients of dry masa flours in two neighbor rural communities located in México. Infants
and children who received the soybean fortified masa flour grew 49% more than counterparts fed
with regular diet. Clinical tests showed that subjects consuming the fortified/enriched tortillas
improved their hair, nails and skin conditions. Pregnant women that consumed fortified tortillas
gave birth to newborns with significantly higher weights and only 3% of the babies had lower
Page 4 of 35
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M - 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1
5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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adopted this technology because the soybean flour affects the organoleptic properties. To our
knowledge there are not investigations aimed to compare functional properties of soybean
proteins in the maize tortilla system. Therefore, the aim of this research was to evaluate the
functionality of five different commercial types of soybean proteins in maize tortillas. The yield
and physical, chemical, sensorial and textural shelf-life during storage at room temperature were
used as the criteria to select the best soybean protein.
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C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M - 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1
5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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MATERIALS AND METHODS
Maize and soybean flour samples
Composite flours were obtained by mixing commercial nixtamalized maize flour (Control
treatment, Maseca Premium Plus, high yield dry masa flour supplied with gums and emulsifiers)
with about 6% (6 g per each 100 g of nixtamalized maize flour) of four different defatted
soybean flours (Industrial de Alimentos, SBF1; GAF 120, SBF2; ADM, SBF3 and Ragasa,
SBF4) or 4% (4 g per each 100 g of nixtamalized maize flour) soybean concentrate (Nutrigrains
Industries, SBC). The percent soybean flour/concentrate added was fixed at the above levels so
as to obtain 25-30% higher protein content in the composite flour'. In order to produce the
dough or masa, one part of the dry masa flour was hydrated with about 1.3 parts water (water
was adjusted after a pre-evaluation of the hydration performance of the composite flours). The
level of protein enrichment allowed an increase in essential amino acid content in at least 30%
compared with the control treatment.
Rheological, chemical and functional characterization of flour.
The viscoamylograph properties of the control and soybean enriched dry masa flours were
evaluated with the Rapid Visco Analyzer (RVA, StarchMaster2, Perten Instruments) according
to Fernández-Muñoz et al (2011). The soybean products were characterized for moisture (44-
15A -AACC, 2000-), crude protein (AACC 46-13 -AACC, 2000-), pH and electrical
d i i (B hT M HI 2550 H I ) F A i Ni (FAN
Page 6 of 35
C e r e a l C h e m i s
t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M - 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1
5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v
e r s i o n m a y d i f f e r .
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Solubility Index (NSI), and percentage coagulation at high temperature were determined
according to Cheftel et al (1989). Foam Capacity (FC) was calculated as recommended by
Haque and Kito (1983). The complete amino acid profile was obtained following the AOAC
Official Method 982.30 (AOAC 1980).
Tortilla production at pilot plant facility
Composite flours were blended with tap water which was added based in subjective evaluations
of the handling properties of the resulting dough or masa. 1 kg of regular or composite flours was
blended with 1.15 to 1.45 L of water for 3.5 min in a Hobart mixer (5L N-50 Planetary Mixer)
equipped with a hook attachment. The resulting masa was sheeted and formed into 30 g tortilla
circles and continuously baked in a three-tier baking oven for 68 seconds (Manufacturas C&D
Industriales, Monterrey, NL, Mexico). The average weight of tortillas obtained was 27 grams
and the surface temperature of each stage at the oven was set to 230, 270 and 250°C. Once the
water absorption was tuned for each experimental flour, trials were performed in 2 kg batches.
The baked tortillas were allowed to cool down on a counter for 30 min at room temperature
(25±2°C) and immediately packaged in polyethylene bags at room temperature for textural,
color, sensorial and chemical evaluations.
Chemical and sensory properties of maize tortillas
Tortilla moisture was determined using the oven method (44-15A, -AACC, 2000), crude protein
i h h i Kj ld hl d (46 30 AACC 2000) d d f f h i
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C e r e a l C h e m i s
t r y J o u r n a l " F i r s t L o o k " p a p e r • h
t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M - 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v
e r s i o n m a y d i f f e r .
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Tortilla quality during storage
Tortillas were stored at room temperature and at day 0, 1, 2 and 5 each treatment was evaluated
for luminosity, texture and rollability. Color was obtained using a colorimeter (CR 300, Minolta,
Osaka Japan) at least in three samples and three different spots of the same sample. Lighting
conditions used for color test were the same (performed under the same lamp and at the same
distance). Texture changes were determined as the maximum force required for penetration using
a cylindrical probe attached to a Texturometer (TA XT 2i, Stable Micro System) whereas
rollability was subjectively rated by rolling tortillas around a 1 cm wooden dowel. The rating
scale employed was 1 (no cracks; very flexible) to 5 (break immediately; cannot be rolled)
(Serna-Saldívar 2012).
Integral evaluation of studied parameters
In order to perform an overall assessment of the maize tortillas obtained with flour enriched with
soybean products, an evaluation with the key parameters for producers and consumers (color,
texture and sensorial components as well as process yield) was completed. For each parameter, a
percentage of the total score was assigned using an empirical approach (experience in
nixtamalized maize tortilla production and consumption). Briefly: yield is a trait very important
to tortilla producers (30/100 weight) as well as sensorial characteristics (30/100). The latter in
turn are of paramount importance for consumers, as well as all texture-related traits (rollability
and texture 30/100 both). Luminosity is also a very important tortilla characteristic, mainly for
Page 8 of 35
C e r e a l C h e m i s
t r y J o u r n a l " F i r s t L o o k " p a p e r • h
t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M - 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o
r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v
e r s i o n m a y d i f f e r .
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basis). Once the evaluation frame was set, values from 1 to 6 were assigned to each treatment
according to statistical differences obtained after the different analyses. One was the best and 6
the worst grade. An overall score was estimated, being the highest value the worst assessment
from consumers and producers viewpoints.
Statistical analysis
Chemical, functional, textural properties of flours as well as process yield for each triplicated
treatment were analyzed with ANOVA (Analysis of Variance) procedures. Differences among
means were calculated with Tukey’s tests (alpha=0.05). Correlation analysis was made with
triplicate results for the five samples used for protein enrichment. The statistical software used
was Statistix (v.9).
Page 9 of 35
C e r e a l C h e m i s
t r y J o u r n a l " F i r s t L o o k " p a p e r • h
t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e
w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o
r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v
e r s i o n m a y d i f f e r .
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RESULTS AND DISCUSSION
Chemical, rheological and functional properties of composited flours
Differences among chemical and functional properties of soybean products are depicted in Table
I. pH values ranged from 6.6 to 6.8 for all defatted soybean flours and the concentrate. Moisture
was also similar, except for SBF 3 which contained 14.3%. The lowest and highest protein
contents of 42.4 and 67.1% were for the SBF1 and the SBC, respectively. The last could be then
classified as a concentrate because it contained a protein higher than 65 and lower than 70%. The
rest of the soybean products are categorized as defatted meals obtained after desolventization in a
desolventizer-toaster (DT; Mustakas 1971; Riaz 2006).
Urease activity values differed among the soybean protein sources. This assay is related to the
protein denaturation due to heat treatments applied during the process. SBF1 and SBF2 had
values below 0.2 whereas SBF3 and SBF4 values higher than 2.0 indicating that the first group
received more heat during processing. The urease activity is closely related to PDI so SBF 3 and
4 had the highest PDI values indicating an insufficient heating or diminished protein
denaturation. According with Smith (1977), urease activity of 0.3 or less suggests that the
product (despites a slight urease activity) has received an adequate heat treatment for the
deactivation of antinutritional factors. Different PDI materials were purposely tested because
their less denatured proteins have different functionalities especially during masa formation and
assuming that the inhibitors were destroyed during the subsequent tortilla baking step.
Page 10 of 35
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h
t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e
w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o
r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v
e r s i o n m a y d i f f e r .
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regularly has PDI values of 90, followed by defatted soy flour with 70. On the other hand,
enzyme inactive soy flours, regularly have PDI values of 20 to 30 (Riaz 2006). High PDI
ingredients are more soluble, because the thermal treatment alters the hydrophilicity /
hydrophobicity ratio of the protein surface (Riaz 2006). High solubility in soybean protein is a
desirable trait, because of the required mixing of the ingredients with water. Furthermore, soluble
proteins are easier to incorporate in food systems (Jideani 2011). Water solubility (WS) as the
percentage of solids solubilized in water (under standard conditions) compared to the total initial
amount of the evaluated sample, were 25.7, 28.4, 50.3, 47.6 and 37.0 for SBF1, SBF2, SBF3,
SBF4 and SBC respectively, results consistent with PDI values. According to Jideani (2011)
defatting increases protein solubility, being this property also influenced by the hydrophilicity /
hydrophobicity balance which in turn depends on amino acid composition (Moure et al 2006).
Other functional property related with hydration is water absorption index (WAI), which can be
described as the ability of the meal to retain water against gravity. According to Moure et al
(2006), the retained water includes: bound, hydrodynamic, capillary and physically entrapped
and mainly depends on the amino acid composition and exposure of amino acid residues. WAI
improves with the number of charged amino acid residues (Kuntz and Kauzmann 1974).
Electrical conductivity (data not showed) was higher for SBC (3.6 ±0.12 mS/cm) compared with
the rest of the soy products (2.0±0.0 to 2.6±0.0 mS/cm) indicating differences in the
concentration of ions and electrical conductive particles and in some degree to the capacity of the
material to absorb water WAI for the array of soy products (Table I) indicated that the highest
Page 11 of 35
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h
t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e
w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o
r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v
e r s i o n m a y d i f f e r .
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absorption. This could be the reason why SBF1 and SBF2, with a high thermal treatment
(depicted by their low PDI value), were comparatively rated with higher WAI.
Regarding FAN, the highest values were associated to SBF3, SBF4 and SBC (Table I). FAN
determines the nitrogen readily available for microorganisms and chemical reactions and gives
an indication of degree of protein hydrolysis. Porter and Jones (2003) determined FAN for 22
soybean flours obtaining an average of 1.22 ± 0.11 mg/g. This mean value was similar to the
observed in SBF3 and SBF4 that received a mild heat treatment during processing (after
industrial oil extraction). According to Porter and Jones (2003), FAN concentration varies with
the soybean used for flour production and maturity of seeds at harvest and moisture of beans in
the different stages of production and storage.
One of the most important parameters within this evaluation was indeed the protein amino acid
profile of the composite flour used for tortilla production (Table II). As planned, addition of
approximately 6% soybean flours and 4% SBC increased 25 to 30% the protein content. All
cereal-based products had lysine as the most limiting amino acid followed by tryptophan. On the
other hand, all composite flours and tortillas have adequate amounts of sulfur containing amino
acids due to the high levels of maize proteins known to have excess levels of these amino acids
(Table II). Regarding lysine, maize contains around 2.0 to 2.5 g of this amino acid per 100 g
protein. With the addition of SBF 1 to 4 and SBC, lysine content increased 56 to 95% (Table II).
The essential amino acid profile of the enriched tortilla flours was thus greatly improved due to
Page 12 of 35
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h
t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e
w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o
r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v
e r s i o n m a y d i f f e r .
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treatments, as previously described, lysine was the limiting amino acid followed by tryptophan.
All other essential amino acids exceeded the requirements of protein for a 2 to 5 year-old infant
according to FAO and WHO recommendations (WHO, 2007). In the specific case of lysine,
control met the 50% of the daily requirement, whereas soybean enriched treatments between
67.8 to 69.1%. This important improvement in protein quality greatly improves protein
digestibility corrected essential amino acid scores, nitrogen retention and growth of both
experimental animals and humans (Amaya-Guerra et al 2004, 2006; Bressani et al 1974, 1979,
Bressani 2008; Chavez and Chavez 2004; Del Valle and Perez Villaseñor 1974; Serna-Saldívar
and Amaya-Guerra 2008).
Similar results were reported with other protein enrichment protocols and the direct addition of
essential and limiting amino acids. For example, Lecuona-Villanueva et al (2012) developed an
enriched nixtamalized flour by directly adding 0.2% lysine and 0.03% tryptophan yielding
similar levels to the obtained in this research. This type of fortification has been widely studied
because of the amino acid balance and the influence of this factor in animal and human growth,
nevertheless, the use of soybean protein is ideally suited to counteract deficiencies because it
contains at least 15 times more lysine and tryptophan compared to maize and other cereals.
Besides, soybean products are generally sold at low cost and have high market availability.
The typical viscoamylograph curves of the control and enriched masa flours are depicted in Fig.
1. According to Almeida-Dominguez et al (1996) the viscoamylograph behavior is an effective
Page 13 of 35
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h
t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e
w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o
r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v
e r s i o n m a y d i f f e r .
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enriched with the SBC. The soy concentrate depicted a different behavior compared with the rest
of treatments because the composite flour needed a higher pasting time and lower pasting
temperature and peak and set back viscosities. The pasting temperature is related to the energy
required to cook the sample indicating that the SBC somehow interfered with the starch
gelatinization phenomena. Other interesting difference is that the flour enriched with SBC had
more stability to prolonged thermic process (95°C), because of the steady viscosity observed
during this stage of the assay.
Peak viscosity, an indicator of the water-binding capacity (Perten 2011) of starch granules, was
lower for the flour enriched with the SBC (Fig. 1). Interestingly, the SBC had the highest water
absorption index among all the evaluated soybean materials (8.34 versus an average of 4.8 for all
the defatted soybean flours).
One parameter also useful in food processing is the holding strength of the evaluated material or
hot paste viscosity. This indicates the ability of the sample to withstand the heat and shear stress.
All samples depicted a similar shear thinning except for flours containing SBF1 and SBC, which
performed better due to the lower viscosity decrease during the heat holding period of the assay.
All enriched flours depicted a similar set back viscosity, rheological parameter related to
synaeresis of starch upon cooling of the cooked starchy material (Sandhu and Singh 2007) and
thus to the rate of starch retrogradation and loss of tortilla texture during storage (Fig. 1). The
highest and lowest final viscosities were observed for flours containing SBF1 and SBC,
Page 14 of 35
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h
t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0
1 5
a p e r h a s b e e n p e e r r e v i e
w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o
r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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In Table III, results of chemical composition and textural characteristics of maize tortillas are
depicted. All tortillas contained moistures between 37.4 to 50.5% related to the water adjustment
made during the masa forming process. Interestingly, the SBF 3 and 4 enriched tortillas
contained lower moisture compared to the rest of the treatments likely due to their high PDI
values. The flour enriched with SBF1 and SBF2 produced tortillas that with practically the same
moisture compared to the control tortillas. Bressani (2008) reported an average tortilla moisture
content of 47.8%, similar to SBF1, SBF2, SBC and control (Table III).
As expected, all soybean enriched tortillas contained higher protein compared to the control. The
protein content for the enriched tortillas ranged from 11 to 14% compared to 9.17% observed in
control tortillas. The tortillas enriched with SBF4 and SBF2 contained slightly higher protein
content compared to the rest of the enriched counterparts. According to Bressani (2008) the
average protein expressed on dry basis for white and yellow tortillas was 10.3 and 10.6%
respectively. According to Serna-Saldívar and Amaya-Guerra (2008), the protein quality of raw
corn and its tortillas is considered low, because of the deficiencies on lysine and tryptophan.
One parameter used for texture and quality assessment during storage is rollability and bending
tests (Serna-Saldívar 2012). These assays detect changes in tortilla texture throughout storage,
being the latter an objective measurement that can be correlated with subjective information
obtained from rollability and flexibility test scores. Suhendro et al (1998) devised a tortilla
bending technique and a rollability test sensitive to changes in texture. Maximum forces required
g
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0
1 5
a p e r h a s b e e n p e e r r e v i e
w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o
r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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penetration force (8.95 N) whereas the SBF3 and SBC tortillas the softest with only 5.9 and 6.8
N, respectively (Fig. 2). Interestingly, these tortillas were produced with the high PDI soybean
flours. Thus, the low heat treated soybean flours favored the texture of stored tortillas.
Texture and color during storage of control and soybean-enriched tortillas.
The different tortilla systems had dissimilar changes in color and texture during storage. In
terms of color, the luminosity or L values dropped due to addition of soybean products (Fig. 3).
The tortilla enriched with SBF4 which had the highest FAN showed the lowest luminosity
whereas the rest of the fortified tortillas had similar values compared to the control. All tortillas,
except the ones enriched with SBF4, gradually decreased L values during storage. The rate of L
value loss was similar among treatments. Green et al (1977) described changes in color of
fortified tortillas when full-fat soy flour was used above 12%. Figueroa et al (2001) observed
that with only 4% of defatted soy flour in tortilla (+ vitamin addition) yielded a product with
different color compared to the control.
In terms of tortilla texture, the maximum force changed from day 0 to the fifth day of storage at
room temperature (Fig. 2). All fresh tortillas (day 0) had an average penetration force of
4.48±0.51 N and 7.58±1.12 N at the end of storage (day 5). Tortillas enriched with SBF3 were
softer or required less penetration force (5.93 N) compared to counterparts enriched with SBF1
(8.95 N). These results clearly indicate that tortillas lose texture during storage due to starch
retrogradation and that addition of various soybean sources affected differently the texture of
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0
1 5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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In Fig. 4 results for sensory evaluation of tortillas are displayed. Five parameters were evaluated:
color, taste, odor, texture, as well as general tortilla acceptance. The highest the value, the lowest
acceptability. Regarding color, all enriched tortillas were evaluated better than the control. SBF4,
the treatment with lower luminosity (Fig. 3) was one of the best evaluated by the panelists. SBF1
was one of the best with respect to taste and SBF2 regarding odor. The last two materials (SBF1
and SBF2) were also the lower in PDI and urease activity values (Table I). SBF3 tortillas scored
low in terms of taste, odor and overall acceptability, being this soy enriched tortilla similar to
SBF4 in terms of chemical and functional properties (Table I).
Changes in tortilla yield due to the addition of the various types of soybean proteins.
Tortilla yield is depicted in Fig. 5. The best treatments were the control and the SBF1 enriched
tortilla followed by tortillas enriched with SBC5, SBF2, SBF3 and SBF4, respectively. It is
important to mention that the control commercial flour is a high yielding due to its low moisture
content and supplementation with hydrocolloids and emulsifiers. These results indicate that the
SBF4 (high PDI) was unable to retain as much water during masa mixing and tortilla baking
operations compared with other sources as SBF1 and SBF2 with reduced PDI. Addition of high
PDI soybean flours SBF3 and SBF4 significantly reduced tortilla yields compared to the control
and SBF1 enriched counterparts.
Overall evaluation of five different soybean proteins used in nixtamalized maize
tortillas production.
C e r e a l C h e m i
s t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0
1 5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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percentages of the total score (100%), were assigned to each factor according to the relative
importance of each one from the consumers and producers viewpoints. In this evaluation, the
lowest the overall score, the highest the desirability of the tortilla system. The control and SBF1
enriched tortillas were the best overall evaluated products. Thus, the SBF1 with a PDI of 23.17%
yielded higher amounts of enhanced protein quality tortillas. These tortillas had similar sensory
properties, texture throughout storage and color compared to the control.
Correlation among the physicochemical characteristics of soy products and the most
important tortilla parameters
In order to further analyze the role and functionality of soy-products within a nixtamalized
tortilla production system, a correlation assessment was performed and the coefficients are
summarized in Table V. The most significant relationships were between yield-related
parameters (tortilla and masa moisture) and PDI, urease activity and WS. These connections
resulted negative and highly significant. The lower the PDI, Urease Activity and WS, the higher
tortilla final yield. A similar correlation was found among acceptability and thermic treatment
associated parameters as WS. The lower the WS, the better the general acceptability.
Different to yield and acceptability, textural related parameters as rollability and texture force
were positively correlated with PDI and WS. The higher the PDI and WS, the best tortillas in
terms of textural properties during storage. Regarding texture, mild treatments in soy products
are convenient for their use as supplements for tortillas. In terms of texture, the best rated soy
C e r e a l C h e m i
s t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0
1 5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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CONCLUSIONS
Functional properties of five different soybean commercial products (SBF1 to SBF4 and SBC)
within maize tortilla system were fully evaluated in terms of texture, color, sensorial
acceptability and nutrimental profile, the most important parameters for producers and
consumers of this Latin-American traditional food. Tortillas were enriched with 3% of protein
weight from the assessed soybean ingredients in order to increase between 25 to 30% protein
content in dry masa flour, yielding composite flours with an upgraded essential amino acid
profile (double lysine and tryptophan) compared to the 100% nixtamalized maize counterpart.
These flours depicted differences in viscoamylographic properties compared to the control
(lower viscosity and setback for SBC). Soybean ingredients were characterized and two of them
showed high UA and PDI, whereas a third one had a relatively protein concentration compared
with soybean flours (>60%). Tortillas produced using this enrichment strategy depicted similar
chemical properties, differences in color and slightly differences in the texture after five days of
storage. The best tortillas in terms of texture (texturometer and rollability) were the produced
with SBF3 and SBF4 (materials with high UA and PDI) but were the worst in yield evaluation.
The SBF4 was the poorest in luminosity evaluation. Interestingly these differences were not
detected by the sensorial evaluation panel. Correlation analysis depicted a significant negative
association between yield-related parameters (tortilla and masa moisture) and PDI, urease
activity and WS. Regarding texture, mild heat treated soy products are convenient for their use as
tortillas supplements After an integral assessment tortillas produced with SBF1 were the best
C e r e a l C h e m i s t r y J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0
1 5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n
b u t h a s n o t y e t b e e n c o p y e d i t e d o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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ACKNOWLEDGMENTS
The authors would like to acknowledge USSEC for providing the array of soybean samples and
sponsoring this research.
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TABLES
Table I. Chemical and functional properties of the soybean products used to enrich maize
tortillasa
Soy
Product
Chemical Functional
pH
Moisture
(%)
Protein
(%, db)
Urease
Activity
FAN
(mg/g)
Reducing
sugars
(mg/g)
Water
Absorptio
n Index
(WAI)
PDI
(%)
Fat
Absorption
Index
(FAI)
SBF 1 6.67±0.007 b 8.28±0.11b 42.4±0.17 e 0.10 ± 0.01e 0.62±0.01 d 81.0±0.40 c 5.38±0.02b 23.17 ± 0.11 d 2.5 ± 0.02 d
SBF 2 6.63±0.000 bc 7.93±0.16b 49.6±1.90 c 0.15 ± 0.01 d 0.59±0.01 d 5.0±0.40 d 4.8±0.25c 25.34 ± 1.55 d 2.94 ± 0.03 b
SBF 3 6.48±0.026d 14.25±0.8a 58.6±0.44b 2.25 ± 0.01a 1.23± 0.02 c 88.0±0.70 a 4.02±0.03d 75.45 ± 1.62 a 2.65 ± 0.05 c
SBF 4 6.78±0.023a 3.58±0.07c 45.4±0.10 d 2.20 ± 0.03 b 1.40± 0.03 b 85.0±0.7 b 4.29±0.07d 66.96 ± 1.51 b 3.14 ± 0.03 a
SBC 6.56±0.057 c 4.11±0.03c 67.1±0.20 a 0.40 ± 0.01 c 1.87±0.04 a 7.0±0.40 d 8.34±0.08a 52.03 ± 0.45 c 2.7 ± 0.09 c
a Data in the table are the average of at least three replicas ± standard deviation. Means with different letter(s) within columns are
statistically different (P < 0.05).
C e r e a l C h e m
i s t r y J o u r n a l " F i r s t L o o k " p a p e r •
h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n b u t h a s n o t y e t b e e n c o p y e d i t e d
o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d
v e r s i o n m a y d i f f e r .
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Table II. Essential amino acid content for the mixes used for tortilla production
Essential Amino Acid
Amino acid (g/100g protein)
Control
Flour Enriched with
SBF1 SBF2 SBF3 SBF4 SBC
Threonine 3.62 3.81 3.81 3.82 3.82 3.74
Valine 4.83 4.97 4.95 4.94 4.98 4.93
Methionine + Cysteine 3.98 2.97 2.97 2.97 2.97 2.97
Isoleucine 3.62 4.01 4.00 3.97 4.02 3.99
Leucine 13.03 11.94 11.94 11.89 11.93 11.94
Phenylalanine+Tyrosine 7.60 5.67 5.67 5.67 5.67 5.67
Lysine 2.90 3.93 3.94 3.97 4.01 3.94
Histidine 2.90 2.89 2.89 2.88 2.90 2.90
Tryptophan 0.72 0.98 0.99 0.96 0.94 0.93
Amino acid scorea 49.9 67.8 68.0 68.4 69.1 68.0
a Limiting amino acid was lysine for all treatments considering a requirement for infants of 5.8 g lysine/100 g protein.
C e r e a l C h e m
i s t r y J o u r n a l " F i r s t L o o k " p a p e r •
h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n b u t h a s n o t y e t b e e n c o p y e d i t e d
o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d
v e r s i o n m a y d i f f e r .
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Table III. Chemical composition and textural characteristics of nixtamalized maize tortillas
enriched with different soy productsa.
a Data in the table are the average of at least three replicas ± standard deviation. Means with different letter(s) within columns are
statistically different (P < 0.05).
b Crude protein (nitrogen x 6.25) and fat values are expressed on dry matter basis.
c Subjective scale from 1 to 5, where 1 indicates an excellent rollability and 5 a poor performance during this test.
Tortilla sample Moisture (%)
Crude
Proteinb (%)
Crude Fatb
(%)
Cold
rollability
at five days
of storage
c
Maximum force
at five days of
storage
(Newtons, N)
Control 50.5±1.4 a 9.17± 0.45 c 3.71±0.01 a 2.7±0.14 a 8.23±0.22 ab
SBF 1 48.6±0.9 ab 11.52±0.01 bc 4.82±0.10 a 3.6±0.07 a 8.95±0.17 a
SBF 2 48.3±0.4 ab 13.10±1.09 ab 5.07±1.32 a 3.4±0.28 a 8.36±1.02 ab
SBF 3 37.4±1.3 c 10.71±0.31 bc 1.24±0.12 b 3.0±1.84 a 5.93±1.07 b
SBF 4 37.6±1.4 c 14.22±0.96 a 1.15±0.15 b 2.2±1.41 a 7.15±0.47 ab
SBC 45.6±0.3 b 10.73±1.70 bc 1.53±0.06 b 3.3±0.49 a 6.84±0.65 ab
C e r e a l C h e m
i s t r y J o u r n a l " F i r s t L o o k " p a p e r •
h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M
- 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
a p e r h a s b e e n p e e r r e v i e w e d a n d a c c e p t e d f o r p u b l i c a t i o n b u t h a s n o t y e t b e e n c o p y e d i t e d
o r p r o o f r e a d .
T h e f i n a l p u b l i s h e d
v e r s i o n m a y d i f f e r .
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Table IV. Nixtamalized maize tortillas overall evaluationa
Treatment
Parameter
Overall
ScoreRollability Luminosity
Sensory
Evaluation
Yield
Texture
(maximum
force)
Control 1 1 1 1 2 120
SBF1 1 1 1 1 3 140
SBF2 2 1 1 2 2 160
SBF3 1 1 1 3 1 160
SBF4 1 2 1 4 2 220
SBC 1 1 1 2 2 150
Weight of each
parameter (%)
10 10 30 30 20 100
a Values are in a scale from 1 to 6 where 1 is the best and 6 the worst grade. The same value was assigned to treatments with no
significant difference according with Tukey’s analysis (alpha
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Chuck-Hernández et al
()
()
()
()
()
()
(
)
(
)
(
)
1.00
, % 0.72 1.00
() 0.4 1.00
()
0.5 0.74 0.12 1.00
()
0.63 0.64 0.55 1.00
()
0.6 0.31 0.70 1.00
()
0.20 0.32 0.47 0.61 0.32 0.10 1.00
()
0.10 0.42 0.43 0.6 0.20 0.23 1.00
0.46 0.04 0.65 0.3 0.56 0.16 1.00
0.64 0.14 0.53 0.03 0.4 0.76 0.67 1.00
0.04 0.03 0.2 0.56 0.21 0.20 0.73 0.6 0.73 0.50 1.00
0.6 0.41 0.12 0.66 0.40 0.71 0.64 0.71 0.6 1.00
0.42 0.15 0.54 0.31 0.5 0.3 0.7 1.00
0.26 0.07 0.37 0.42 0.4 0.25 0.72 1.00
(
)
0.23 0.12 0.44 0.46 0.37 0.2 0.7 0.73 0.63 1.00
(
)
0.12 0.6 0.17 0.74 0.01 0.45 0.75 0.55 0.4 0.35 0.56 0.71 0.50 1.00
(
)
0.04 0.50 0.44 0.75 0.06 0.41 0.52 0.57 0.5 0.64 0.71 0.62 1.00
Table V. Correlation table with the main physicochemical characteristics of soy products and final parameters of maize tortillaa
1
a Correlation coefficients significant with P
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FIGURES
Fig. 1. Effect of soybean enrichment on viscoamylographic properties of water suspensions composited flours used for tortilla
production.
45
50
55
60
65
70
75
80
85
90
95
0
500
1000
1500
2000
2500
0 100 200 300 400 500 600 700 800 900 1000
T e m p e r a t u r e
( ° C )
V i s c o s i t y ( R V U )
Time (seg)
CONTROL SBF1 SBF2 SBF3
SBF4 SBC Temperature
C e r e a l C h e m i s t r y
J o u r n a l " F i r s t L o o k " p a p e r • h t t p : / / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M - 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
T h i s p a p e r h a s b e e n p e e r r e v i e w e d
a n d a c c e p t e d f o r p u b l i c a t i o n b u t
h a s n o t y e t b e e n c o p y e d i t e d o r p r
o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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Chuck-Hernández et al
Fig. 2. Effect of soybean enrichment on luminosity of tortillas stored for five days at room temperature.
68
70
72
74
76
78
80
82
84
0 1 2 5
L u m i n o s i t y ( L )
Storage (days)
Control
SBF1
SBF2
SBF3
SBF4
SBC
C e r e a l C h e m i s t r y
J o u r n a l " F i r s t L o o k " p a p e r • h t t p :
/ / d x . d o i . o r g / 1 0 . 1
0 9 4 / C C H E M - 0 7 - 1 4 - 0 1 5 4 - R • p o s t e d 0 2 / 2 3 / 2 0 1 5
T h i s p a p e r h a s b e e n p e e r r e v i e w e d
a n d a c c e p t e d f o r p u b l i c a t i o n b u t
h a s n o t y e t b e e n c o p y e d i t e d o r p r
o o f r e a d .
T h e f i n a l p u b l i s h e d v e r s i o n m a y d i f f e r .
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Chuck-Hernández et al
Fig. 4. Sensory evaluation of enriched tortillas produced with five different soy products.
A subjective scale was used during the sensorial trials, where