Plant Foods for Human Nutrition 62: 31–37, 2007.


c 2007 Springer Science+Business Media, Inc.
DOI: 10.1007/s11130-006-0039-z
Nutritional Properties of Quality Protein Maize and Chickpea Extruded Based Weaning Food
J. MILÁN-CARRILLO,1, 2 C. VALDÉZ-ALARCÓN,1 R. GUTIÉRREZ-DORADO,1
O. G. CÁRDENAS-VALENZUELA,1 R. MORA-ESCOBEDO,3 J. A. GARZÓN-TIZNADO2, 4 &
C. REYES-MORENO1, 2, ∗
1 Maestrı́a en Ciencia y Tecnologı́a de Alimentos, Facultad de Ciencias Quı́mico – Biológicas, Universidad Autónoma de Sinaloa (FCQB-UAS), México;
2 Programa Regional del Noroeste para el Doctorado en Biotecnologı́a, FCQB-UAS, México; 3 Escuela Nacional de Ciencias Biológicas, Instituto
Politécnico Nacional, México; 4 Centro de Investigaciones Regionales del Noroeste, Instituto Nacional de Investigaciones Forestales, Agrı́colas y
Pecuarias, Valle de Culiacán, Sinaloa, México (∗ author for correspondence Present address: Lichis No. 1986, Colonia La Campiña, 80 060 Culiacán,
Sinaloa, México; e-mail: creyes@uas.uasnet.mx)
Published online: 23 January 2007
Abstract. Malnutrition is one of the major causes of morbidity and mor-
tality among young children in most of the developing countries. To mini-
mize the adversities of malnutrition, low-cost infant supplementary foods
have been developed and are being supplied to the needy through state-
sponsored nutrition intervention programmers. The present study had two
objectives: to determine the best combination of nixtamalized extruded
quality protein maize (NEMF) and extruded chickpea (ECF) flours for
producing a weaning food, and to evaluate the nutritional properties of
the optimized NEMF/ECF mixture and the weaning food. The NEMF
and ECF were produced applying combinations of extrusion tempera-
ture/screw speed of 79.4◦ C/73.5 rpm, and 150.5◦ C/190.5 rpm, respec-
tively. Response surface methodology was applied to determine the op-
timum combination NEMF/ECF; the experimental design generated 11
assays. Mixtures from each assay were evaluated for true protein (TP) and
available lysine (AL). Each one of 11 mixtures were used for preparing
11 weaning foods which were sensory evaluated for acceptability (A).
The best combination of NEMF/ECF for producing a weaning food was
NEMF = 21.2%/ ECF = 78.8 %. This mixture had a global desirabil-
ity (D) of 0.93; it contained 20.07% proteins (DM), 5.70% lipids (DM),
and 71.14% carbohydrates (DM); its essential amino acids (EAA) profile
satisfactorily covered the EAA requirements for children 2–5 years old,
except for Trp. The weaning food prepared with the optimized mixture
had high protein quality and digestibility and could be used to support the
growth of infants.
Key words: Nixtamalized xtruded maize flour, Extruded chickpea flour,
Nutritional properties, Response surface methodology, Weaning food
Introduction
Malnutrition is one of the major causes of morbidity and
mortality among young children in most of the develop-
ing countries. To minimize the adversities of malnutrition,
low-cost infant supplementary foods have been developed
and are being supplied to the needy through state-sponsored
nutrition intervention programmers. Various food process-
ing techniques have the potential to enhance the nutrient
bioavailability, nutrient density, food safety, storage sta-
bility, palatability, and convenience of supplemental foods
suitable for infant mixtures. These technologies suitable
for weaning foods include roasting, germination, milling,
baking, cooking, drying, fermentation, and extrusion [1].
Extrusion is a technology classified as a high tempera-
ture/short time process; it offers numerous advantages in-
31
cluding versatility, high productivity, low operating cost,
energy efficiency, high quality of resulting products and an
improvement in digestibility and biological value of pro-
teins [2, 3].
Maize is third in importance, after wheat and rice, as
a staple human food; it has a moderately low content of
protein that has an unfavorable amino acid composition.
There is a marked deficiency of Lys and Trp, a moderate
deficiency of Ile, and an excess of Leu, a constellation con-
tributing to development of pellagra or florid kwashiorkor
[4]. Quality protein maize (QPM) is similar to normal maize
in agronomic characteristics but contains greater amounts
(70–100%) of Lys and Trp [5]. In México, researchers
from International Maize and Wheat Improvement Center
(CIMMyT) and the National Research Institute for Forestry,
Agriculture and Livestock (INIFAP), have successfully de-
veloped 26 hybrids and cultivars of QPM that are sim-
ilar in yield and other agronomic properties to normal
maize [6].
Chickpeas are a good source of proteins and carbohy-
drates, possessing important quantities of some vitamins
(thiamin, niacin), minerals (Ca, P, Fe, Mg, K) and un-
saturated fatty acids (oleic, linoleic). However, chickpeas
have several undesirable attributes, such as long cooking
times, enzyme inhibitors, phytates, flatus factors, and phe-
nolic compounds, which must be removed or eliminated
for effective utilization; furthermore, chickpeas proteins
are deficient in sulfur amino acids [7, 8]. Some researchers
[9] have reported that phenolic compounds could be con-
sidered as bioactive compounds due to their antioxidant
capacity.
When mixed together, the proteins of maize and chick-
peas complement one another to produce a protein of a bet-
ter quality by providing to each other significant amounts of
the respective limiting amino acids. The present study had
two objectives: (1) To determine the best combination of
nixtamalized extruded quality protein maize and extruded
chickpea flours for producing an infant food, and (2) To
evaluate the nutritional properties of the optimized mixture
and the weaning food.
32
Materials and Methods Table 1. Experimental designa used to obtain differents combina-
tions of nixtamalized extruded quality protein maize flour/extruded
chickpea flour for producing weaning foods, and experimental results
Materials for response variables

Quality protein maize (Zea mays L) V-537, and chickpea Component (%)c Response variabled
(Cicer arietinum L) Blanco Sinaloa 92 varieties were ob- Assayb NEMF(X1 ) ECF(X2 ) TP (Y1 ) AL (Y2 ) A (Y3 )
tained from the INIFAP, Culiacán, Experimental Station,
Sinaloa, México. 1 100 0 8.21 3.5 2.2
2 50 50 13.81 4.8 3.1
3 0 100 18.17 6.1 3.7
4 75 25 10.22 4.2 2.5
Methods 5 25 75 17.29 5.4 3.5
6 50 50 13.76 4.8 2.7
Preparation of nixtamalized extruded maize flour (NEMF). 7 100 0 8.06 3.5 2.0
NEMF was prepared according procedure proposed by 8 0 100 17.79 6.1 3.4
Reyes-Moreno et al. [10]. Five hundred gram lots of qual- 9 50 50 13.59 4.8 2.8
10 100 0 8.32 3.5 2.2
ity protein maize (QPM) kernels were placed in a domes- 11 0 100 18.14 6.1 3.5
tic blender at low velocity to break the kernels, the grits
a Lattice simplex with two componentes; 11 assays.
obtained were ground in the same blender to obtain grits b Does not corresponded to order of processing.
(1–2 mm) and fine powder. Maize grits were separated from c NEMF: Nixtamalized extruded maize flour, ECF: Extruded chickpea
fine powder using a sieve. Maize grits were mixed with lime flour.
(0.24 g Ca(OH)2 /100 g maize) and water to reach a moisture d TP: True protein (%, DM), AL: Available lysine (g/100 g protein),

content of 28 g/100 g. Each lot was packed in a polyethy- A: Acceptability

lene bag and stored at 4◦ C for 12 h. Before extrusion, the
grits were tempered at 25◦ C for 8 h. The extrusion was car-
ried out on a single screw laboratory extruder Model 20 DN
(CW Brabender Instruments, Inc, NJ, USA) with a 19 mm
screw-diameter. Extrusion temperature (ET) was defined as
temperature at die end barrel. The extruder was operated
at ET = 79.4◦ C and screw speed (SS) of 73.5 rpm. QPM
extrudates were cooled and dried (25◦ C, RH = 65%) for 1
day and milled to pass an 80-US mesh (0.180 mm) screen,
packed in plastic bags, and stored at 4◦ C.
Preparation of extruded chickpea flour (ECF). The ECF
was prepared according procedure proposed by Milán-
Carrillo et al. [3]. Lots (500 g) of whole chickpeas were
placed in a domestic blender at low velocity to break the
chickpeas and provide for air separation of seed coats
from cotyledon grits using a domestic fan. Seed coats
were totally eliminated and cotyledon grits were ground
in a domestic blender at medium velocity to obtain small
grits (1–2 mm). Cotyledon grits were adjusted to 26.5 g
H2 0/100 g sample with a salt solution ((0.25 g NaCl +
0.75 g NaHCO3 )/100 ml of distilled water, w/v), and then
the lot was packed in a polyethylene bag and stored at
4◦ C for 12 h. Before extrusion, the cotyledon grits were
tempered at 25◦ C for 8 h. Extrusion was carried out us-
ing a single screw laboratory extruder Model 20 DN with
the characteristics above mentioned. The operation con-
ditions were: Extrusion temperature 150.5◦ C, and screw
speed 190.5 rpm. Chickpea extrudates were cooled, dried,
milled, packed and stored like QPM extrudates to obtain
extruded chickpea flour (ECF).
Preparation of weaning foods. The Table 1 presents dif-
ferent mixtures of NEMF/ECF used for producing wean-
ing foods. Mixtures from each assay were analyzed for
true protein (TP), and available lysine (AL). Each one of
11 mixtures was used as base for preparing 11 weaning
foods; these were sensory evaluated for acceptability (A).
The weaning foods were prepared as follows: each mix-
ture (25 g) was added with sucrose (8 g) and purified water
(80 ml); the suspension was stirred with a glass rod, heated
(90◦ C/8 min), cooled at room temperature (25◦ C) and sen-
sory evaluated. All determinations were made in triplicate.
Proximate composition. The following official meth-
ods of the AOAC [11] were used to determine proxi-
mate composition: drying at 105◦ C for 24 h, for moisture
(method 925.098); incineration at 550◦ C, for ash (method
923.031); defatting in a Soxhlet apparatus with 2:1 chlo-
roform/methanol, for lipids (method 920:39C with minor
modifications); and microKjeldahl for proteins (Nx6.25)
(method 960.52).
True protein (TP). The TP content was determined by
the difference between total nitrogen and nonprotein nitro-
gen (NPN) determined by microkjeldahl (method 960.52,
AOAC) [11]. To convert into true protein content, the
amount of nitrogen given by this difference was multi-
plied by the 6.25 factor. About of 0.5 g of sample, with a
known amount of total nitrogen previously determinated by
microkjeldahl, was added with 20 ml 10% trichloroacetic
acid (TCA) solution and shaken using an orbital mixer at
400 rpm and 25◦ C for 1 h. The insoluble material was re-
moved by centrifugation. The supernatant was made up to

50 ml using distilled water and an aliquot was taken for the
determination of NPN by microkjeldahl. The NPN remains
soluble after the protein precipitation by TCA.
Available lysine (AL). A procedure recommended by
Hurrel et al. [12] was applied. Acid orange 12 dye was
used. The dye binds to lysine residues of protein, which
precipitates at acid pH. Samples (0.5 g) were placed in Er-
lenmeyer glass flasks. In the first step (reading A) 40 ml
of acid orange solution [1.3626 g dye/l in phosphate buffer
(20 g oxalic acid + 3.4 g monobasic potassium phosphate
+ 60 ml acetic acid), pH 1.25] were added to a flask with
the sample. In the second step (reading B), corresponding
to propionylation, 0.2 ml of propionic anhydride and 2 ml
of phosphate buffer were added to another flask with the
sample. Both flasks were shaken at 400 rpm and 25◦ C for
1 h. Aliquots (15 ml) from each flask were centrifuged at
5000 × g and 25◦ C for 15 min; each supernatant fluid was
diluted with distilled water (1:100). The absorbances were
measured using a UV-Visible spectrophotometer (Spec-
tronic 21D Model 1146, Milton Roy, USA) at 475 nm. A
standard curve of lysine was constructed. The amount of ly-
sine was obtained by subtracting reading B from reading A.
In vitro protein digestibility (PD). The methodology pro-
posed by Hsu et al. [13] was used. A multienzyme system
[(8 mg trypsin + 15.5 mg chymotrypsin + 6.5 mg pepti-
dase)/5 ml of distilled water] consisting of porcine pancre-
atic trypsin type IX, bovine pancreatic chymotrypsin type II
and porcine intestinal peptidase grade III (Sigma Chemical
Co, St Luis, MO, USA) was selected. A specific amount of
each sample and distilled water were used to prepare 50 ml
of an aqueous protein suspension (6.25 mg protein/ml) with
pH adjusted to 8.0, while stirring in a 37◦ C water bath. The
multienzyme solution was maintained in an ice bath and
adjusted to pH 8.0. Five milliliters of the multienzyme so-
lution were then added to each protein suspension which
was being stirred at 37◦ C. The pH drop was recorded auto-
matically over a 10 min period using a recording pH meter.
In vitro protein digestibility (PD) was calculated applying
the equation PD = 210.46 – 18.10 X, value X = pH ten
min after adding enzymes to protein suspension [13].
Amino acids analysis. Five to ten mg of each sample
were placed in 2 ml ampoules containing internal standard
(norleucine) and 0.4 ml 6 N HCl. The ampoules were evac-
uated, sealed, and placed in an oven at 110◦ C for 24 h. After
hydrolysis, a 20 µl aliquot of the hydrolisate was dried and
subject to derivatization. Samples for the determination of
cysteine were first oxidized with performic acid at room
temperature for 18 h [14]. The tryptophan content was
determined in a separate analysis. The samples were hy-
drolyzed in polypropylene tubes in 4.2 M KOH containing
1% (w/v) thiodiglycol at 110◦ C for 18 h. After hydrolysis,
the KOH was neutralized with 4.2 M percloric acid. The
supernatant was removed and adjusted to pH 3 with dilute
acetic acid, and a 50 µl aliquot was used for derivitiza-
33
tion. Quantitation was achieved using a Pierce Standard H
amino acid calibration mixture that was supplemented with
tryptophan.
The amino acid analyses were perfomed using the Pico-
Tag system (Waters, Milford, MA). Tryptophan was ana-
lyzed using the conditions described by Hariharan et al.
[15]. A sample of sodium caseinate was utilized as the
control protein. All determinations were made in duplicate.
Calculated protein efficiency ratio (C-PER). The C-
PER’s were calculated using the procedure suggested by
Satterlee et al. [16] and summarized by the AOAC [11].
The procedure was based on utilization of the in vitro pro-
tein digestibility and essential amino acids composition of
the analized sample.
Sensory analysis. Thirty min after cooking, samples
(20 g) of weaning food were served in cups with three-
digits random numbers covered with lids. Sensory analysis
was performed by 20 trained judges. Panel members were
students, faculty and staff of Biological and Chemical Sci-
ences Faculty, Sinaloa State University. Initial screening
and selection of panelist was based on participant inter-
est, taste and odour acuity ability to understand the test
procedures. Panelists were seated in individual booths in a
temperature (25◦ C) and relative humidity (50–60 %) con-
trolled rooms. Samples were rated for acceptability on a
6-points hedonic scale (0 = dislike extremely, 5 = like ex-
tremely) [17]. Sensory test were repeated three times. Data
were analyzed for statistical significance by the Duncan’s
test at the 5% level of probability.
Experimental design. Response surface methodology
(RSM) was applied to determine the optimum combination
of nixtamalized extruded maize flour (NEMF) and extruded
chickpea flour (ECF) for producing a weaning food. A two
component Lattice Simplex mixture design was chosen.
The mixture components considered were X1 = NEMF,
and X2 = ECF. The component proportions were expressed
as fractions of the mixture and the sum (X1 + X2 ) of the
component proportions equaled 100 %. The experimental
design generated 11 assays (Table 1). Mixtures from each
assay were evaluated for true protein (TP) and available
lysine (AL). Each one of 11 mixtures were used as base
for preparing 11 weaning foods; these were sensory evalu-
ated for acceptability (A). TP, AL and A were considered
response variables. The following empirical “black box”
modeling presents the relationships among components and
response variables.
The expression inside the “black box,” Yk, represents
predicted dependent variable (each property, TP, AL, and A,
β 1 , β 2 , and β 12 are corresponding parameter estimates for
34
each linear and cross-product term, respectively, produced Table 2. Regression coefficients and analyses of variance of the
first-order polynomial equations (predictive models) showing the re-
for the prediction models. Multiple regression analysis was
lationships among response variables (Yk ) and components of the
applied and predictive model for each dependent variable mixture variables (X)
were obtained [18].
Optimization. The objective in the optimization process True protein Available Aceptability
was to find a common optimum value for the three depen- Coefficient YTP Lysine YAL YA
dent variables; thus, the desirability method was used [19]. Linear
The three fitted models can be evaluated at any point X = β1 8.20∗ 3.49∗ 2.14∗
(X1 , X2 ) of the experimental zone, and as a result three val- β2 18.04∗ 6.09∗ 3.55∗
ues were predicted for each model, namely Ŷ1 (X ), Ŷ2 (X ), Interactions
β 12 2.64NS −0.02NS 0.28NS
and Ŷ3 (X ). Then each Ŷi (X ) is transformed into a value di
R2 0.998 0.999 0.95
(X), which falls in the range (0, 1) and measures the desir- P≤ 0.0001 0.0001 0.001
ability degree of the response in reference to the optimum
∗ Significant at p ≤ 0.10, NS: No significant.
value intented to be reached. In this research, we wanted
all response variables to be as high as possible. Thus, the
transformation is Predictive Models for Response Variables
⎧
⎪
⎪ 0 if Ŷ1 (X ) ≤ Yi ∗
⎪
⎨ Experimental results for dependent variables at different
Ŷi (X ) − Yi ∗
di (X ) = if Y i ∗ ≤ Ŷ1 (X ) ≤ Yi∗ NEMF/ECF combinations are presented in Table 1. Rela-
⎪
⎪ Y ∗
− Y
⎪ i
⎩ i+ tionships among components of the mixtures and dependent
1 if Ŷi (X ) ≥ Yi ∗ variables through predictive models are shown in Table 2.
The value Yi∗ corresponds to the highest value of all
response variables; therefore, it is the minimum acceptable. Essential Amino Acids (EAA) Profile of the Extruded Flours
Once the three individual desirabilities were calculated,
the next step was to obtain the global desirability for the The EAA composition and their scores of proteins from
three response variables, using the mathematical function extruded flours are given in Table 3 along with those for
of transformation D = (d1 d2 d3 )1/3 . requirements for children 2–5 years old recommended by
Where the ideal optimum value is D = 1; an acceptable FAO/WHO [22]. The contents of EAA such as His, Leu,
value for D can be between 0.6 and 0.8 (0.6 < D < 0.8). and Phe + Tyr, in both flours, sulfur EAA (Met + Cys) and
This acceptable value was found by utilizing the Design Val in NEMF, and Lys, Trp and Thr in ECF, where found
Expert system [18].
Statistical analysis. All results were analyzed using one- Table 3. Nutritional properties of nixtamalized extruded quality protein
way analysis of variance (ANOVA) followed by Duncan’s maize and extruded chickpea flours
multiple range test (p ≤ 0.5) [18].
FAO/WHO/UNU
Requirements
Results and Discussion for children
Propertya NEMFb ECFb 2–5 years old
Properties of Extruded Flours EAA (g/100 g protein)
His 3.51 2.60 1.9
The nixtamalized extruded quality protein maize flour Ile 2.63 2.92 2.8
(NEMF) contained 10.75 g proteins (DM), 5.84 g lipids Leu 9.12 7.15 6.6
(DM), and 81.4 g carbohydrates/100 g (DM) . These re- Lys 4.12 6.58 5.8
Met + Cys 5.03 2.08 2.5
sults are in agreement with those reported by Bello-Pérez Phe + Tyr 7.33 8.14 6.3
et al. [20] for nixtamalized corn flours. The extruded chick- Trp 0.89 1.2 1.1
pea flour (ECF) had higher (p ≤ 0.05) contents of pro- Thr 3.5 3.9 3.4
teins and ashes and lower (p ≤ 0.05) carbohydrates and Val 5.07 3.52 3.5
lipids contents than did NEMF. Khan et al. [20] reported EAA score 71.03 83.20
Limiting EAA Lys Met + Cys
23.1–26.79 g proteins, 3.6–5.2 g lipids, 2.8–3.6 g ashes and In vitro protein digestibility 80.9 82.6
45.5–56.7 g carbohydrates/100 g (DM), in four different C-PER 1.57 1.89
desi and kabulli chickpea varieties. Differences in proxi-
a EAA: Essential amino acids, C-PER: Calculated protein efficiency
mate composition of NEMF and ECF would be expect to
ratio.
affect the physicochemical and sensory properties of the b NEMF: Nixtamalized extruded maize flour, ECF: Extruded chickpea
weaning foods. flour.

TP
Individual
AL
A 0.96
0 0.25
0.92
0.69
Desirability
0.46
0.23
NEMF = Nixtamalized extruded maize flour
ECF = Extruded chickpea flour
0
NEMF (%) 0 25
ECF (%) 100 75
Figure 1. (a) Individual desirability (d), and (b) global desirability (D = 0.93) for the optimized
NEMF/ECF mixture for preparing a weaning food.
to be higher than those of the FAO/WHO recommended
pattern [22]. The other EAA, Thr in NEMF and Ile and Val
in ECF, were at levels comparable to the reference pattern.
The contents of Lys in NEMF, and sulfur EAA (Met + Cys)
in ECF, seem to be the limiting EAA.
Optimization
The objective in the optimization process was to find the
NEMF/ECF combination that resulted in the optimum val-
ues for the three dependent variables (TP, AL, A). To predict
the dependent variables at the optimum value, the com-
plete models were considered in the desirability function
[19]. The common optimum values for the three dependent
variables were obtained at a global desirability value of
0.93, as a result of combination (21.2 g NEMF + 78.8 g
ECF)/100 g optimized mixture (Fig. 1). This combination
was recognized as “optimized mixture” which was used as
basic ingredient for preparing an weaning food.
Properties of the Optimized NEMF/ECF Mixture
The optimized NEMF/ECF mixture contained 20.07 g pro-
teins, 5.7 g lipids, 71.14 g carbohydrates, and 3.09 g
35
(a)
0.99
0.71
0.93 Global
0.50 0.75 1.0
(b)
D= 0.93
NEMF = 21.2%
ECF = 78.9%
50 100
75
50 25 0
ashes/100 g mixture, DM (Table 4). This optimized mix-
ture satisfactorily covered the specifications of a Joint
FAO/WHO Committee that recommended minimum lev-
els of 16.1 g protein, 6.0 g fat, and 375 Cal/100 g of sample
[23]. The EAA content of optimized mixture were found to
be higher than the requirements for children 2–5 years old
recommended by FAO/WHO [22], except for Trp (Table 4).
The optimized mixture had in vitro protein digestibility and
C-PER of 84.5% and 1.85, respectively (Table 4). Serna-
Saldı́var et al. [24] carried out studies on nutritive value
of cereal-legume mixtures evaluating in vitro protein di-
gestibilities, amino acids profile, C-PER, and PER; they
reported, for wheat breads fortified with deffated soybean
and sesame meals, in vitro protein digestibilities and C-
PER’s of 83.1–84.87% and 1.22–1.35, respectively. These
researchers concluded than PER’s obtained by the use of
in vitro digestibilities and amino acids underestimated the
values obtained from rats; however, the absolutes differ-
ences between treatments were similar, indicating that the
C-PER technique predicted the same differences observed
in the rat bioassay. Therefore, it is recommended to use the
in vitro techniques as fast, accurate indicators of protein
digestibilities and PER’s.
36
Table 4. Chemical composition and nutritional properties of optimized be used to support the growth of infants in developing
NEMF/ECF mixture
countries.
FAO/WHO/UNU
Optimized requirements for Acknowledgements
mixture of children 2–
Propertya NEMF/ECFb 5 years old
This research was supported by the Consejo Estatal de
Chemical composition (%, DM) Ciencia y Tecnologı́a – Estado de Sinaloa and Universi-
Proteins 20.07 dad Autónoma de Sinaloa, México.
Lipids 5.70
Carbohydrates 71.14
Minerals 3.09 References
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