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CAPÍTULO 1

Effect of Moisture and Temperature on the Functional Properties of

Composite Flour Extrudates from Beans (Phaseolus vulgaris) and

Nixtamalized Corn (Zea mays).

ABSTRACT

Beans have high protein content. Bean and corn flours can complement each other

with essential amino acids. There has been little research on the production of

snacks with bean–corn composite flours. The aim of this study was to obtain a

bean–corn snack with high protein content for low-income families in Mexico.

Bean–corn composite flours (60/40) were extruded. The effect of temperature and

moisture during extrusion on the end quality of the product was analyzed. The

expansion index, apparent density, water solubility index, water absorption index

and the initial viscosity were measured and were significantly (p < 0.05) higher

when bean–corn flour was extruded in high temperature and low moisture

conditions. The best bean–corn snack was obtained in extrusion conditions of 190

ºC and 14.5% moisture. The results show that bean proteins can be complemented

by corn proteins to obtain highly valuable protein flour. Extrusion is an alternative

processing method for obtaining snack products with high protein content for low-

income families.

Palabras claves: Harinas de frijol, harinas de maíz nixtamalizado, extrusión

1.1 Introduction

Beans (Phaseolus vulgaris) are one of the main protein sources for many low-

income families in developing countries. The soluble fiber present in beans has

beneficial effects in the prevention of cardiac diseases. Long processing times and

the presence of antinutritional compounds in beans limit their use. The inactivation

of trypsin inhibitors (TI) and lectins is very important in bean processing (Chang

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and Satterlee, 1982; Buera et al., 1984; Rackis et al., 1986; Liener, 1986). Even

the low TI activity found in processed bean foods increases pancreatic tumors in

rats (Noah et al., 1980; Yavelow et al., 1982; Gumbmann et al., 1985; Wang and

Chang, 1988; Wang et al., 1988).

The sulfur amino acids are limiting in dry beans, while Lys and Try are limiting in

corn. Beans and corn can complement each other with essential amino acids

(Marshal et al., 1982; Eicher and Satterlee, 1988).

Nixtamalization is a traditional alkali treatment in which corn is precooked with

Ca(OH)2, conditioned for 6 – 18 hrs, washed and stone-ground to produce masa,

which is then processed to produce different products (Gomez et al., 1991). Tortilla

and other Mexican corn products are made from nixtamalized corn (Serna-Saldivar

et al., 1998). Nixtamalized corn retains most of the germ, aleurone and some

pericarp layers (Paredes-Lopez and Saharopulos 1982; Gomez et al., 1989).

During nixtamalization different molecular and granular forms of starch occur, not

only because of the partial gelatinization of starch, but also due to retrogradation

(Paredes-Lopez and Saharopulos 1982; Gomez et al., 1989; Gomez et al., 1990).

It has also been shown that starch birefringence decreases by 47% during

commercial corn nixtamalization, but lower gelatinization and less birefringence

loss (5–15%) may be achieved with less severe nixtamalization conditions

(Pflugfelder et al., 1988; Gomez et al., 1989). Nixtamalized corn flour has better

nutritional properties than untreated corn flour. Nixtamalization increases the

Lys/Iso ratio, Ca content, and protein digestibility and decreases aflatoxin

contamination (Trejo-Gonzalez et al., 1982; Paredes-Lopez and Saharopulos,

1982; Rodriguez, 1995; Bryant et al., 1997; Fernandez-Muñoz et al., 2001;

Zazueta et al., 2002). Nixtamalized corn flours can be used for bread and snack

production, but are most often used for tortillas.

Extrusion is cheap, versatile, highly productive and consumes low amounts of

energy (Harper, 1981). Confectionary products, baby foods, snacks, ready-to-eat

breakfast cereals and pet foods are extruded products (Akdogan, 1999). In

general, extrusion products are made out of pure starch or high starch content

cereals, since starch gelatinization provides texture and structure to the end-

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product (Wang et al., 1989; Billiarderis et al., 1980). Extrusion of bean flour is

complicated, because of its relatively low starch and high protein content. An

expansion index ranging between 1.5 and 2 may be reached with soybean

extrusion. The highest expansion index was reached at 18% moisture (Zasypkin

and Tung-Ching, 1998). In general, the expansion ratio increases with higher

extrusion moisture and temperature (Wang et al., 1999). The degree of expansion

also increases with higher pressure, which is produced with higher extrusion

moisture. Bulk density can be increased by decreasing moisture content, barrel

temperature and screw speed. The expansion and texture of extrudates is more

complex for products based on more than one component. The multiphase

structure affects the elastic properties of the extrudates (Tsebrenko et al., 1974).

Expansion is reduced at concentrations of 50% of each component (Yuriev et al.,

1995; Zasypkin and Tung-Ching, 1998). The best expansion index is obtained at

concentrations of 80% or more wheat flour and 10% or less of soybean flour.

Extruded blend flours have low sensitivity to moisture content, and therefore the

processing of blended flours may be simpler than that of flour alone (Zasypkin and

Tung-Ching, 1998).

Extruded snacks are quite popular among the Mexican population (Jackson et al.,

2004). An extruded snack made of bean–nixtamalized corn flour may increase the

nutritional value of these products.

The aim of this study was to determine the effect of extrusion parameters on the

functional properties of bean–nixtamalized corn flour.

1.2 Material and Methods

1.2.1 Bean and nixtamal

Whole bean flour (Phaseolus vulgaris L.) cultivar Pinto Villa from the highlands of

Durango, Mexico was used. Beans were grown in spring from 2006. The beans

were milled in a commercial mill (MLI 204, Buehler, Switzerland). Corn (Zea mays

L.) from the cultivar CAFIME grown in 2005 was nixtamalized as described by

Arambula et al. (2001) and Trejo-Gonzalez (1982). The nixtamalized corn was

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milled in a nixtamal-stone mill (Villamex, Guadalajara, Mexico). Bean and

nixtamalized corn (N-corn) flours were mixed in a proportion of 60 and 40%,

respectively.

1.2.3 Extrusion

Extrusion was done with a single screw extruder (CINVESTAV, Queretaro, Mexico)

with a compression of 3:1, a screw diameter of 19 mm, a relation length-diameter

of 20:1 and a die diameter of 3.0 mm. A constant screw speed of 90 rpm (60 Hz)

and a constant feeding speed of 28 rpm were used. The temperature in the third

zone of the extruding zone was varied (150, 160, 170, 180 and 190 °C). The

desired moisture level was adjusted by spraying distilled water onto the bean/N-

corn flour, which was then hand mixed for 15 min and conditioned to 14.5, 15.4,

17.1 or 18.0% moisture for 12 hrs in closed plastic containers at 8°C. Moisture was

determined with the approved method from 44-15A (AACC, 2000). An

experimental central rotary design of second order was used. Samples were

identified as follows: 1 = 142 °C/16.3% H; 2 = 150 °C/14.5% H; 3 = 150 ºC/18% H;

4 = 170 °C/16.3% H; 5 = 170 °C/18.7% H; 6 = 170 °C/16.3% H; 7 = 170 °C/13.8%

H; 8 = 170 °C/16.3% H; 9 = 170 °C/16.3% H; 10 = 170 °C/16.3% H; 11 = 190

°C/18% H; 12 = 190 °C/14.5% H and 13 = 198 °C/16.3% H. Three separate

extrusion runs were carried out for each sample.

Extruded samples were dried at 45 °C for 24 hrs. Samples were ground in a Bühler

CE mill (Bühler, S.P.A., Switzerland) and sieved through a 60 mesh screen. The

samples were kept in sealed glass vials for further analysis.

1.2.4 Expansion index and bulk density

The expansion index (EI) and bulk density (AD) were determined as previously

described (Gujska and Khan, 1990). Results are shown as the mean of ten

repetitions. The quality control index (QCI) of the EI was calculated as follows:

EI QCI EI

σ =

6



where σ = standard deviation of EI.

The experimental data for EI and AD were adjusted to a second order quadratic

model (Myers, 1992): Yi= b0 + b1X1 + b2X2 + b11X12 + b22X2

2 + b12X1X2, where Yi =

the response, X1 = temperature, X2 = moisture and b0, b1, b2, b11, b22 and b12 are the

regression coefficients.

1.2.5 Water absorption and solubility index

The water absorption index (WAI) and water solubility index (WSI) were

determined as described by Anderson et al. (1969). Results are shown as the

mean of three repetitions. An equation model was calculated for WAI and WSI.

The experimental data for WAI and WSI were adjusted to a second order quadratic

model (Myers, 1992): Yi= b0 + b1X1 + b2X2 + b11X12 + b22X22 + b12X1X2, where Yi =

the response, X1 = temperature, X2 = moisture and b0, b1, b2, b11, b22 and b12 are

the regression coefficients.

1.2.6 Hardness

Hardness was determined with the Texture analyzer (TX-XT2i, Stable Microsystem

Co. Ltd., UK), as described by Gujska et al. (1996). Results are shown as the

mean of three repetitions. The experimental data for the hardness was adjusted toa second order quadratic model (Myers, 1992): Yi= b0 + b1X1 + b2X2 + b11X1

2 +

b22X22 + b12X1X2, where Yi = the response, X1 = temperature, X2 = moisture and b0,

b1, b2, b11, b22 and b12 are the regression coefficients.

1.2.7 Scanning electron microscopy

Scanning electron microscopy was carried out as described by Gomez et al.

(1991).

1.2.8 X-Ray diffraction analysis

To determine the structural changes in nixtamal and bean starch during extrusion,

X-ray diffraction patterns of samples and crystallinity were determined as described

by Gomez et al. (1990). The loss of starch crystallinity during extrusion can be

7



seen as an indirect measure of starch gelatinization. This method was used to

determine crystallinity loss and indirectly to reach some conclusions about the

degree of gelatinization of extruded samples.

1.2.9 Statistical analysis

The experimental design and the data analysis were carried out with a response

surface methodology with Design Expert ® Software (Stat-Ease, Inc., Minneapolis,

MN, USA).

1.3 Results and Discussion

1.3.1 Expansion Index

The EI of an extrudate is one of the most important physical characteristics.

Expansion affects tenderness, fragility and density (Conway and Anderson, 1973).

Temperature and moisture had an effect (p <0.05) on the EI of extruded bean–corn

flour. Differences (p < 0.05) were observed between sample 12 and 13, 12 and 11,

12 and 7 and 12 and 8 (Table 1). The EI was higher (p < 0.05) in sample 12 than

samples 7, 8 and 11. Moisture had an effect (p < 0.05) on EI in samples extruded

at 190 °C. At temperatures as low as 142 °C EI was the same as at 198 °C at

equivalent moisture levels. The results obtained were comparable to the EI

obtained by other authors with corn or wheat flour complemented with bean

proteins or soybean flour, respectively (Gujska and Khan, 1991; Zasypkin and

Tung-Ching, 1998). Temperatures of 142 and 190 °C in bean–nixtamalized corn

blends had no effect (p > 0.05) on EI, in contrast to other studies (Balandran-

Quintero et al., 1998), where extrusion of bean flour alone had a lower EI at 140 °C

than at 180 °C. Extrusion of bean flour alone gives a high EI at temperatures above

170 °C (Balandran-Quintero et al., 1998), while our results showed that the bean–

nixtamalized corn blend produced a similar high EI at temperatures as low as 142

°C. An EI of 2.3 was obtained for extruded corn; this value was not obtained with

bean–corn flour in our investigations, since starch is the primary cause of

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expansion, and the starch content in bean flour is lower than the starch content in

corn flour. The lower EI obtained in bean–corn extrudates may be explained by the

differences in the internal structures of the extrudates. Corn extrudates have larger

and smoother air cells than extrudates with high protein content. Extrudates with

high protein content have small cells with wrinkled walls (Gujska and Khan, 1991).

Moisture content did not have an effect (p > 0.05) on EI at equivalent

temperatures. These results contrast with findings in soybean flour (Zasypkin and

Tung-Ching, 1998).

Findings in soybean flour, wheat flour, sorghum and cowpea show that individual

flours have a higher EI than the blends (Falcone and Phillips, 1998; Zasypkin and

Tung-Ching, 1998). This may also be the case with pure bean flour, although this

was not investigated in this paper.

Analysis of variance indicated that the model was not acceptable (p > 0.05; r 2 =

0.59; CV = 4.95) and cannot be used to predict values for EI (Fig. 1).

).**1066.9()*1056.3()*1080.3()*07.0()*01.0(62.0 42326 H T H T H T EI

−−−

•−•+•−+++=

The lower the QCI the better the product similarity. The best QCI was obtained for

sample 7 with an extrusion temperature of 170 °C and 13.8% moisture. Low

extrusion moisture at 170 °C produces extrudates with more similar EIs. Extrusion

at 150 °C and 14.5% moisture is also acceptable for processing of bean–corn flour,

due to the lower energy consumption and similarity among the end-products (Table

1).

1.3.2 Bulk density

In contrast to other findings (Colonna et al., 1989; Kokini et al., 1992), bulk density

was affected (p < 0.001) by temperature and moisture conditions during extrusion

(Table 1). Samples 1, 2, 3, 7 and 9 had a significantly higher (p < 0.05) bulk

density than samples 11, 12 and 13. A lower AD indicates a better structure of the

extrudates (Gujska and Khan, 1991). Moisture content had no effect (p > 0.05) on

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the AD of samples extruded at 190 °C. Extrusion at 190 °C or more produced a

lower AD than extrusion at 150 °C or lower. Samples 1 and 13 indicate that AD

decreased (p < 0.05) due to the high extrusion temperature rather than moisture

content, although an extrusion temperature above 190 °C had no effect (p > 0.05)

on EI. It is recommended to carry out extrusion at 190 °C to obtain a low bulk

density. Bulk density did not increase (p > 0.05) in extrudates produced at 170 °C

and different moisture levels, compared to extrusion at 144 °C. Moisture content

showed no effect (p > 0.05) on AD at 150 °C extrusion. The bulk density obtained

between 142 and 170 °C is comparable to the bulk density of corn flour extruded at

132 °C by other authors, and lower if extruded at 190 °C or more (Gujska and

Khan, 1991). Protein-enriched extrudates have a bulk density of up to 0.60 g/cm

3

(Gujska and Khan, 1991). However, in our study, extruded bean–corn flour did not

reach those values, which indicates a smooth structure of bean–corn extrudates.

Other authors have found that wheat gluten and soy protein can increase

expansion and strength, indicating that protein type and concentration affect EI and

AD (Faubion and Hoseney, 1982). The extruded bean–nixtamalized corn blend

had a lower AD compared with other reported findings for extruded bean flour

alone (Balandran-Quintero et al., 1998).

Analysis of variance indicated that the model was acceptable (p < 0.05; r 2 = 0.86;

CV = 9.86) and could be used to predict values for bulk density (Fig. 1):

)**1059.9()*1018.2()*1033.1()*23.0()*01.0(85.3 42325 H T H T H T AD

−−−

•+•+•−−−+=

1.3.3 Hardness

Hardness was affected (p < 0.05) by extrusion temperature and moisture (Table 1).

Snacks extruded at 170 °C and 16.3% moisture were softer (p < 0.05) than snacks

extruded at 13.8% moisture and 170 °C in two out of three cases. Extrusion at 190

°C or at 198 °C produced softer (p < 0.05) snacks than snacks extruded at 170 °C

and 13.8% moisture. Other authors also found an effect of moisture on the texture

properties of soybean and wheat flour blends (Zasypkin and Tung-Ching, 1998).

10



There was no significant difference (p > 0.05) between the hardness of snacks

produced at temperatures below 150 °C or above 190 °C, nor did moisture show

any significant effect (p > 0.05). Analysis of variance indicated that the model was

acceptable (p < 0.05; r 2 = 0.76; CV = 18.76) and could be used to predict values for

hardness (Fig. 2):

)**016.0()*06.0()*1048.2()*93.4()*11.1(63.137223

H T H T H T H ++•+−−+=−

.

1.3.4 Water absorption index

The WAI was not affected (p > 0.05) either by temperature or by moisture (Table

1). The obtained values are comparable with the WAI of corn flour (Balandran-

Quintero, 1998; Gujska and Khan, 1991), indicating that bean flour has no

significant effect (p > 0.05) on WAI. This is contrary to reports that bean protein

had a negative effect on the WAI (Quintero et al., 1998; Gujska and Khan, 1991),

although these same authors found an increase in WAI by adding high protein

fractions from beans to extrudates, and explained these differences as being due

to the different type of proteins used. Other authors (Anderson et al., 1969, 1982;

Balandran-Quintero et al., 1998) found that temperature and moisture had an effect

(p < 0.05) on the WAI of bean flour alone, which contrasts with our findings, wherethe WAI of bean–nixtamalized flour blends were not affected (p > 0.05) by

extrusion temperature, indicating good water-holding capacity (Colonna et al.,

1989; Kokini et al., 1992). Analysis of variance indicated that the model was

acceptable (p < 0.05; r 2 = 0.77; CV = 5.39) and could be used to predict values for

the water absorption index (Fig. 3):

)**1035.6()*05.0()*1048.5()*43.0()*07.0(52.1 3225 H T H T H T WAI

−−

•−+•+−+= .

1.3.5 Water solubility index

No significant differences (p > 0.05) were found among the WSI of the different

samples (Table 1), indicating that starch depolymerization did not take place at

high temperatures. The WSI values ranged from 9.9 to 15.8, and were lower than

those reported by other authors (Anderson, 1982; Balandran-Quintero et al., 1998;

11



Gujska and Khan, 1991). These authors also found that high protein extrudates

have a lower WSI than corn extrudates and that the WSI increases with increasing

extrusion temperature of bean flour. Gujska and Khan (1991) indicated that the

WSI is low due to greater shear degradation of starch during extrusion at low

moisture. In this study, moisture was no higher than 18%. Analysis of variance

indicated that the model was acceptable (p < 0.05; r 2 = 0.76; CV = 8.36) and could

be used to predict values for WSI (Fig. 3):

).**04.0()*11.0()*1088.2()*53.3()*74.0(14.77 224 H T H T H T WSI −+•−++−=

−

1.3.6 Scanning electron microscopy

The size and shape of starch granules vary depending on the biological origin of

the plant (Svegmark and Hermansson, 1993). The biochemistry of the chloroplast

or the amyloplast and the physiology of the plant determine the form of the starch

granules (Badenhuizen, 1969). Most bean starch granules are >20 µm (Fig. 4A),

being larger than corn and rice starch granules and comparable with wheat and

potato starch granules (Lim et al., 1992). The granular size of bean starch varies

from 20 to 50 µm. Bean starch granules are oval in shape, similar to a bean kernel.

Nixtamalized corn flour contains small starch granules of about 15 µm, as well assome damaged and swollen granules, probably due to partial gelatinization and

endogenous enzyme degradation during nixtamalization (Fig. 4B). These are not

found after wet-milling of corn without nixtamalization (Singh and Johnston, 2002).

Nixtamalization may inhibit gelatinization by amylase–calcium interactions (Robles

et al., 1998). Contrary to other findings (Gomez et al., 1991) no released starch

granules were observed after milling of nixtamal into flour. Corn starch granule size

was comparable to previous reports (Singh and Johnston, 2002). Lim et al. (1992)

determined the average granule diameter of corn starch as 14.3 µm. The corn

starch granules are angular-shaped. In the case of nixtamal, the individual

granules form large clusters; similar findings were reported by other authors

(Gomez et al., 1989, 1991). Nixtamalization causes granules to swell, but little

12



other modification. Figure 4C shows an amorphous starch structure, with no sign of

intact native granules, after extrusion.

1.3.7 Crystallinity

Bean flour shows the highest crystallinity followed by nixtamalized corn. Nixtamal

has a lower crystallinity than bean, because the starch is partially gelatinized

through the nixtamal process (Fig. 5), although starch crystallinity recovers after

nixtamalization through amylase and amylopectin retrogradation (Robles et al.,

1998). Nixtamalization may even increase the crystallinity of starch, when nixtamal

processing takes place below the gelatinization temperature (Donovan et al.,

1983). The variation in crystallinity between different species and cultivars depends

on the amylase/amylopectin ratio, size and shape of the starch granules and

presence or absence of lipids (Bai, 1997; Chang and Lui, 1991; Fan and Marks,

1998; Gudmundsson and Eliasson, 1990). Large granules have a higher amount of

amylase than small starch granules (Pan and Jane, 2000). The crystallinity of

extruded bean–nixtamalized corn flour was 50.2 to 69.5% lower than bean flour.

The results show that extrusion destroys the crystalline structure of starch granules

in bean and nixtamal flour, although some crystallinity is not lost. The extrusion of

bean–nixtamal flours partially gelatinizes starch, while some of the starch granules

retain their native structure and therefore their crystallinity. Native starch can act

as fiber and have a positive effect on digestion of the extruded products. Native

granular starch is one of four forms in which resistant starch occurs (Englyst et al.,

1992; Haralampu, 2000; Niba, 2002). The fermentation products of native and

resistant starch, such as butyrate, which are obtained in the large intestine, may

have various health benefits. These products may lower colorectal cancer risks

(Englyst et al., 1992; Escarpa et al., 1996; Langkilde et al., 1998; Topping and

Clifton, 2001; Niba, 2002). Products containing high levels of resistant starch may

be qualified as functional foods (Johnson and Gee, 1996). Crystallinity did not

affect the hardness of the extruded product (Fig. 5; Table 1).

13



1.4 Conclusions

High temperature extrusion above 170 °C did not increase (p > 0.05) the expansion

index compared to extrusion at 142 °C. Blending of bean flour and nixtamalized

corn flour gave an acceptable expansion index and functional properties

comparable to a corn flour extrudate. The results indicate that extrusion should not

be carried out at temperatures above 170 °C, since the best quality control index

was obtained at this temperature, indicating better similarity of the end product

between batches. Temperatures above 190 °C had no effect (p > 0.05) on EI, WAI,

WSI and hardness, but resulted in lower (p < 0.05) density than extrusion at

temperatures below 150 °C. Starch granules from bean and nixtamal differ in form

and crystallinity. Bean–nixtamalized corn flour extrudates could be used in food

applications with an acceptable expansion index in the temperature and moisture

ranges studied. However, other extrusion variables must be changed, such as a

lower screw speed, to obtain higher density extrudates that may be used as feed in

aquaculture. The results show that starch partly gelatinizes during nixtamalization,

while some granules show enzyme degradation. On the other hand, total starch

gelatinization can be obtained through the extrusion of bean–corn flours.

Acknowledgement: This work was made possible by the financial support of the

Secretaría de Agricultura, Ganadería, Desarrollo Rural, Pesca y Alimentación

(SAGARPA), Consejo Nacional de Ciencia y Tecnología (CONACyT).

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