Received: 2 January 2020 Revised: 22 April 2020 Accepted: 23 April 2020

DOI: 10.1002/leg3.42

ORIGINAL RESEARCH ARTICLE

Formulation and characterization of cookies prepared from the

composite flour of germinated kidney bean, chickpea, and
wheat

Mandeep Singh Sibian | Charanjit Singh Riar

Department of Food Engineering and

Technology, Sant Longowal Institute of Abstract
Engineering and Technology, Longowal, India The objective of this study was to optimize the formulation of cookies from the com-
Correspondence posite flour of germinated kidney bean, chickpea, and wheat using response surface
Mandeep Singh Sibian, Department of Food methodology (RSM). Snap force, spread ratio, and overall acceptability served as
Engineering and Technology, Sant Longowal
Institute of Engineering and Technology, responses for the optimization of cookie formulation. Optimization and validation of
Longowal, India. central composite rotatable design (CCRD) of RSM concluded the feasibility of using
Email: [email protected]
19.11 g of germinated kidney bean flour, 31.19 g of germinated chickpea flour, and
50.00 g of germinated wheat flour, per 100 g of flour composition for cookie prepa-
ration. Characterization of the novel formulated product was done by analyzing vari-
ous attributes of flour and cookies. Optimized composite flour formulation (OCFF)
exhibited appropriate functional and pasting characteristics required for cookie prep-
aration. Optimized composite flour cookies (OCFCs) had a higher amount of protein
(12.32 ± 0.11), fats (22.57 ± 0.23), and crude fiber (5.64 ± 0.02) content as compared
with ungerminated wheat flour cookies (UWFCs). in vitro digestibility (carbohydrate
and protein) was significantly higher in OCFCs owing to the utilization of geminated
grain's flour. Amino acid content of germinated grains enhanced the total essential
amino acids in OCFCs. Shelf life of the formulated product was acceptable for up to
90 days when stored at 25
C in aluminum-laminated sealed bags.

KEYWORDS

composite flour, cookies, germination, in vitro digestibility

1 | I N T RO D UC TI O N Diet-related chronic disease and malnutrition give rise to the con-

cept of novel food processing techniques, of which composite flour is
Wheat flour is an ideal ingredient for various food product formula- one of the effective solutions. Composite flour enriched with func-
tions; therefore, it is widely used for the production of bakery and tional components like tubers, starches, legumes, cereals (Bourré
confectionery products (Diana, Mirela, & Jianu, 2007). The suitability et al., 2019; Noorfarahzilah et al., 2014), and multigrain premixes
of wheat flour in bakery and confectionery products could be attrib- (Kumar, Sharma, Khan, Govindaraj, & Semwal, 2015) reduced the risk
uted to its gluten content. Out of few problems associated with the of diet-related disease and allergies (Adebowale, Adegoke, Sanni,
use of wheat, scarcity of wheat in certain regions and gluten-related Adegunwa, & Fetuga, 2012; Mepba, Eboh, & Nwaojigwa, 2005). Vari-
allergies are common. ous researchers found the suitability of composite flour for the

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium,
provided the original work is properly cited.
© 2020 The Authors. Legume Science published by Wiley Periodicals LLC.

Legume Science. 2020;1–12. wileyonlinelibrary.com/journal/legumescience 1

2 SIBIAN AND RIAR

preparation of bakery products (Gômez, Oliete, Rosell, Pando, & rotatable design (CCRD) was chosen. Preliminary studies have shown
Fernandez, 2008; Ribotta, Arnulphi, Leôn, & Anôn, 2005). Recent that replacing wheat more than 50% with germinated legumes was
trend has also shown the utilization of processed flours or compo- not feasible for production of cookies from composite flour con-
nents that could enhance the nutritional quality of a product. taining germinated legumes. Based on trial runs, the upper and lower
Processing of flour may include germination of grains (Nkhata, Ayua, range for germinated kidney bean flour was selected between 15%
Kamau, & Shingiro, 2018), fortification (Stabnikova, Antoniuk, and 25%, whereas the range for germinated chickpea flour was kept
Stabnikov, & Arsen'eva, 2019), and fermentation (Bourré et al., 2019) between 25% and 35%. CCRD was segregated into three different
of cereal or legumes. Germination is the one of best and inexpensive parts, that is, factorial design (two levels), axial points (outside core),
method to enhance the nutritional qualities and functional character- and center points. RSM statistical software (design expert 10) dis-
istic of grains. Germination allows predigesting of grain's components tributed the design into 13 experiments with four factorial points,
and enhanced free amino acid, vitamins, and functional characteristics five replicates (center points), and four axial points. The CCRD vari-
(Hallén, Ibanoglu, & Ainsworth, 2004). ables and responses for different formulations are as shown in
In the present research, supplementation with germinated legume Table 1. Experiments were carried out in random order to minimize
flours was adopted to formulate a product with high nutrition and the effect of unexplained variability owing to extraneous factors
digestibility. Final formulation was characterized for its nutritional, in (Myers & Montgomery, 2002).
vitro digestibility, amino acid profile, texture profile, and shelf life. Result of responses was analyzed using regression analysis by
fitting a suitable model, that is, second-order polynomial equation:

2 | MATERIALS AND METHODS X

n X
n X
n
Y = β0 + βi xi + βii x2i + βij xi xij ,
i=1 i=1 i6¼j = 1

2.1 | Raw material

where β0 is the value of response at the center points (0, 0); xi and xij
Wheat (PBW-550), kidney bean (light speckled kidney beans), and are the variables of design; βi, βii, and βij are regression coefficients;
chickpea GPF-2 (GF-89-36) were procured from certified seed and n is the number of variables.
agency. Selected grains were germinated as per the method described
by Arora, Jood, Khetarpaul, and Goyal (2009). The sprouts were rinsed
in water after germination, dried initially at 80
C for 15 min to arrest 2.2.2 | Preparation of cookies
the enzyme activity, and then finally dried at 55–60
C to moisture
content of 8.00 ± 1% (db). Dried sprouts and ungerminated wheat Cookies were prepared using a traditional method as described by
grains (PBW-550) were ground separately in lab grinder to fine pow- Chauhan, Saxena, and Singh (2015). Ingredients used were composite
der in the form of flour and passed through 60 mesh sieves (US size flour (proportions were kept according to values given by experimen-
60 mesh = 250 μm). tal design), 100 g; grounded sugar, 40 g; sodium bicarbonate, 1.0 g;
sodium chloride, 1.0 g; bakery shortening, 50 g; skim milk powder,
20 g; and water, 20 ml. A premix of flour, milk powder, and sodium
2.2 | Experimental design bicarbonate was prepared separately. Dough prepared was formed
into sheet of approximately 0.5-cm thickness, and then, a circular
2.2.1 | Selection of independent variables and mold was used to cut the dough sheets. Baking was carried out in bak-
responses ing oven (conventional baking oven, Continental India) at 170
C for
around 15 min. After 15 min, cookies were allowed to cool down at
Selected germinated legume flours other than germinated wheat room temperature. Cookies were packed in aluminum-laminated
flour were selected as independent variables. Response surface sealed bags for further analysis.
methodology (RSM) was used as optimization technique, which has
been proven as appropriate tool in the optimization of various bak-
ery products like sponge cakes (Chaiya, Pongsawatmanit, & Prin- 2.2.3 | Data analysis and estimation of responses
yawiwatkul, 2014), gluten-free bread (Sanchez, Osella, &
Torre, 2004), cassava cake (Gan et al., 2007), and chocolate cake Regression analysis of the experiments was carried out by stepwise
(Moscatto, Borsato, Bona, Oliveira, & Hauly, 2006). Responses like variable selection and backward elimination techniques. Numerical
spread ratio, snap force, and overall acceptability were selected by technique and mapping of responses were used for maximization and
putting a hypothesis that with the incorporation of germinated minimization of polynomial fitting. This was performed by the use of
grains, these functions could be related to specific composition to fit Stat-Ease Design expert software (Ver. 10.0) developed by Stat-Ease
regression equation, which describes the quality composition Inc., Minneapolis. The response surfaces and three-dimensional (3D)
responses. To serve the purpose of optimization and to evaluate the contour graphs were plotted using two function variables. All variables
effect of selected variables on the responses, central composite with significant levels of p ≤ 0.05 were included in a model, and
SIBIAN AND RIAR 3

TABLE 1 Central composite design arrangement and responses for optimization of composite flour with germination legume substitution

Variables (coded) Variables (actual) Responses

Experiment no. x1 x2 x1 x2 Y1 Y2 Y3
1 1.000 1.000 25.00 35.00 6.20 51.24 4.22
2 −1.414 0.000 12.93 30.00 6.30 38.74 3.93
3 0.000 0.000 20.00 30.00 6.40 42.18 4.10
4 −1.000 1.000 15.00 35.00 6.30 43.74 3.93
5 1.000 −1.000 25.00 25.00 6.13 56.17 3.85
6 0.000 0.000 20.00 30.00 6.35 42.12 3.90
7 1.414 0.000 27.07 30.00 6.16 48.26 3.75
8 −1.000 −1.000 15.00 25.00 6.08 48.15 4.22
9 0.000 0.000 20.00 30.00 6.42 45.76 3.92
10 0.000 0.000 20.00 30.00 6.38 43.50 4.05
11 0.000 0.000 20.00 30.00 6.40 41.36 3.98
12 0.000 1.414 20.00 37.07 6.21 58.03 4.75
13 0.000 −1.414 20.00 22.93 6.05 69.88 4.73

Note: x1, kidney bean; x2, chickpea; Y1, spread ratio; Y2, snap force; Y3, overall acceptability.

coefficient of determination was evaluated (R2) to observe the accu- 2.2.6 | Overall acceptability
racy of the model. Samples were estimated for quality characteristics
like snap force (N), spread ratio, and overall sensory attributes. Values A descriptive sensory analysis was carried out by preparing the chart
reported were average of triplicate determinations. and setting out the sensory characteristics as per ISO 13299:2016
procedure, using semitrained analysts to observe the sensory score of
cookies. Overall acceptability was reported finally by calculating the
2.2.4 | Spread ratio average of reported sensory attributes (Gajera, Kapopara, &
Patel, 2010).
Spread ratio was measured as ratio of diameter to thickness of the
cookies (Zoulias, Piknis, & Oreopoulou, 2000). Average value of tripli-
cate measurements was reported, and ratio was observed as 2.3 | Characterization of formulated bakery product

Diameter of cookie ðmmÞ 2.3.1 | Functional properties of flours

Spread ratio = :
Thickness of cookie ðmmÞ

Functional properties of flours were observed by analyzing the water

absorption capacity, oil absorption capacity, sedimentation value,
foaming capacity, and emulsification capacity. Water absorption
2.2.5 | Snap force capacity was done by the method of Yamazaki (1953). Oil absorption
capacity was estimated by the method of Lin, Humbert, and
A three-point bend test was used to estimate the snap force required Sosulski (1974). Sedimentation value was estimated by calculating as
to break down the cookie (Chauhan et al., 2015). For this purpose, TA. the swelling power of flour by using ICC 116/1 standard method
XT2i Stable Micro System texture profile analyzer (Stable (Zeleny's method). Foaming capacity (%) was estimated as per the
microsystem, England) was used with three-point bend rig arrange- method described by Mizubuti, Junior, Souza, da Silva, and Ida (2000).
ments. Mode of measurement was in compression with units in new- Emulsification capacity was calculated by the method of Naczk,
ton (N). The measurements were carried out under following test Diosady, and Rubin (1985). The emulsification capacity (EC) was
conditions. Pretest speed was 1.5 mm/s, test speed was kept at expressed as ml of oil emulsified by 1.0 g of the sample.
2.0 mm/s, posttest speeds were kept at 10.0 mm/s, distance of probe
was maintained at 30 mm, automatic trigger type force was 20 g, and
method settings were maintained by adjusting data acquisition rate at 2.3.2 | Pasting properties of flours
200 pps. Graph was observed between force (N) and time (T). The
peak force required to break down the cookies was reported as snap Pasting properties of different flours were estimated using Rapid
force (Sindhuja, Sudha, & Rahim, 2005). Results were observed in Visco Analyzer (RVA Tecmaster, Perten, Australia), using standard
triplicates. testing profile-1. Appropriate sample around 5 g as described in
4 SIBIAN AND RIAR

manual was taken and mixed with 25 ml of water. The mixture was were estimated from force to deformation curve of compression test
stirred for 10 s at 960 and then 160 rpm. Initially, temp–time combi- (Meullent & Gross, 2007).


nation of 50 C for 1 min was employed, and then, after equilibrium,
temperature raised to 95
C for 5 min. After the procedure, cooling
cycle was carried out by decreasing temperature to 50
C in 3 min. 2.3.5 | Color characteristics of cookies
Results were recorded from graph as peak viscosity (PV), trough vis-
cosity (TV), breakdown value, final viscosity (FV), setback, and pasting Color value of different cookies was estimated by using Hunter Lab
temperature (PT). (HunterLab digital colorimeter, D25M, Reston, Virginia) equipped with
I-type optical sensor (D-25) and DP-9000 processor. L* depicts light-
ness, a indicates redness (+)/greenness (−), and b indicates yellowish
2.3.3 | Proximate analysis and in vitro digestibility of (+)/blueness (−) of the sample. Values of sample were compared with
cookies the standard colored reference tile (Ls = 25.54, as = 28.89, and
bs = 12.03). Cookie samples were placed on specimen port, and the
Moisture content, crude fat content, crude fiber, and ash content values of L*, a*, and b* were recorded.
of cookies prepared from optimized formulation were analyzed
using standard AOAC methods (AOAC, 2005). Total carbohydrate
content was estimated by anthrone method (Ludwig & Gold- 2.3.6 | Amino acid content of cookies
berg, 1956). Protein content of cookies was estimated by the
method of Lowry, Rosebrough, Farr, and Randall (1951). To evalu- Amino acid content of cookies was analyzed using the physiological
ate the protein digestibility, in vitro method suggested by Chavan, kits of gas chromatography–flame ionization detection (Phenomenex,
Chavan, and Kadam (1988) was used. This method is based on the USA). Grounded samples were defatted and then hydrolyzed with
digestion of sample by pepsin and pancreatic enzymes. Digested concentrated HCl. Analysis was performed as instructed in the kit's
and nondigested sample were analyzed for protein by using manual. The GC column used was the ZB-AAA-GC column, which was
method of Lowry et al. (1951). in vitro protein digestibility was cal- provided in the kits, and standard analysis conditions were used, as
culated as proportion of protein digested using following described in the kit's manual. The results obtained were expressed as
relationship: amino acid (g/16 g N).

Total protein− Residual protein

In vitro protein digestibility ð%Þ = × 100:
Total protein 2.3.7 | Storage studies of cookies

For storage studies, both types of cookies were packed separately in

In vitro carbohydrate digestibility was analyzed using the modi- aluminum-laminated sealed bags and stored at 25
C. Different packs
fied method of Modi and Kulkarni (1976). Preweighted defatted sam- of cookie sample (containing 20 cookies each) were prepared for dif-
ple of cookies was treated with 1.5 ml of amyloglucosidase enzyme ferent days (0, 10th, 20th, …, 90th days) of analysis and were labeled
prepared in a 50 mM of sodium acetate buffer (pH 4.5) and incubated accordingly. Microbiological analysis (total plate count) of sample was
at 37
C for 2 h. After incubation, solution was heated to stop enzy- done according to standard AOAC method (AOAC, 2005) on priority
matic reactions. in vitro carbohydrate digestibility was calculated as a basis after opening the packaging, to avoid contamination. Moisture
proportion of residual carbohydrate to total carbohydrates and content, peroxide value, and free fatty acid value of cookies were also
expressed as percentage. analyzed as per the methods of AOCS (1990). Sensory analysis of
cookies was performed at regular intervals to observe the effect of
storage time on cookies.
Total carbohydrate −Residual carbohydrate
IVCD ð%Þ = × 100:
Total carbohydrate

3 | RESULTS AND DISCUSSION

3.1 | Diagnostic checking of fitted model

2.3.4 | Texture profile analysis of cookies
After model reductions, coefficients of regression were observed. Sig-
The three-point bend test was performed by using TA.XT2i Stable nificance of each model was examined by analysis of variance for
Micro System texture profile analyzer (Stable Microsystem, England) observed responses. Model equations were analyzed to observe the
to estimate the texture parameters (hardness, fracturability, and cohe- linear and quadratic effects of variables. Interaction of variable and
siveness) of optimized cookies with similar settings as performed for their effect were observed through regression equations after model
snap test (as discussed in Section 2.2.5). Hardness and cohesiveness reduction. Multiple regression equations at significant levels related to
SIBIAN AND RIAR 5

responses such as spread ratio, snap force, and overall acceptability observed for the regression models to observe the variation in
were reported as follows: responses at different levels of substitution. Regression, lack of fit,
pure error, and residual values were observed as shown in Table 2. F
Spread ratio ðY 1 Þ = 6:3882− 0:0303x1 + 0:0646x
 2 −0:0803x
 1
2
test was conducted to check the adequacy of model. The model F
−0:1298x2 − 0:0376x1 x2 R = 0:969 ,
2 2
values of spread ratio, snap force, and overall acceptability imply that
the model was significant, and there was less chance that a model F
Snap force ðY 2 Þ = 42:9840 + 3:6234x1 −3:2627x
 2 −0:7174x
 1
2
value could occur this large owing to noise. Values of “Prob > F” less
+ 9:5095x2 2 −0:1298x1 x2 R2 = 0:944 ,
than 0.0500 indicated that model terms were significant for each
response. Lack of fit was nonsignificant; therefore, the model was
Overall acceptability ðY 3 Þ = 3:9890−0:0409x1 − 0:0132x
 2 − 0:1339x
 1
2
adequately fit. Regression value of spread ratio and overall acceptabil-
+ 0:3161x2 + 0:1675x1 x2 R = 0:8808 :
2 2

ity was low; therefore, the residual values were closer to 0.

Values for R2 for each model were found valid, which validated
the model fitting. Lack of fit, levels of significance, and analysis of vari-
ance further suggested the fitting and adequacy of model up to a 3.2 | Analysis of variables and responses
desirable significant level. Linear and quadratic effects of the indepen-
dent variable could be estimated from the regression equations after Three-dimensional plots were formed by the responses obtained
model reduction. Negative linear effect of germinated kidney bean after experimental values in the given set of design. Three-surfaced
was observed on spread ratio and overall acceptability, whereas posi- graph provides the predicted data at a very small level of varia-
tive effect was observed on snap force. Germinated chickpea showed tions among independent variables. Spread ratio increased with the
a positive linear effect on spread ratio, whereas negative effect on increase in the germinated chickpea proportion in cookies, which
snap force and overall acceptability was observed. Quadratic effect of could also be attributed to the positive linear effect on spread
germinated kidney bean was observed negative for all responses. Ger- ratio (Figure 1a). A lower increment in the spread ratio was
minated chickpea exhibited negative quadratic effect on spread ratio, observed from the plot when germinated kidney bean proportion
and positive quadratic effect was observed for snap force and overall was increased.
acceptability. Interaction terms of both variables showed significant Snap force dropped at different levels of chickpea substitution,
negative effect on spread ratio and snap force, whereas positive inter- whereas addition of germinated kidney bean caused an increment in
action was observed for overall acceptability. Analysis of variance was the hardness of cookies (Figure 1b). Axial points had higher values of

TABLE 2 Analysis of variance of values for responses for the level of significance for optimization of variable and responses of formulation

Source of variation Sum of squares Degree of freedom Mean square F value

Y1 (spread ratio) Regression 0.1920 5 0.0384 45.10***
Lack of fit 0.0033 3 0.0011 0.308 NS
Pure error 0.0026 4 0.0007 NS
Residual 0.0060 7 0.0009
Total 0.1980 12
Adeq. precision 17.7079
Y2 (snap force) Regression 846.4740 5 169.29 23.76***
Lack of fit 37.8809 3 12.626 4.20 NS
Pure error 12.0027 4 3.001 NS
Residual 49.8837 7 7.126
Total 896.3577 12
Adeq. precision 16.6480
Y3 (overall acceptability) Regression 1.0392 5 0.208 10.341***
Lack of fit 0.1118 3 0.037 5.153 NS
Pure error 0.0289 4 0.007 NS
Residual 0.1407 7 0.020
Total 1.1799 12
Adeq. precision 10.1390

Abbreviation: NS, nonsignificant.

*
p < 0.1.
**
p < 0.05.
***
p < 0.01.
6 SIBIAN AND RIAR

FIGURE 1 Effect of variables on (a) spread ratio, (b) snap force, and (c) overall acceptability

hardness as compared with values closer to the center points. There- purpose. A lower set of deviations from predicted and experimental
fore, the quadratic effect of interaction of both flours was in negative actual values was selected as optimal outcome of design. The RSM-
correlation. based CCRD design was finally obtained for independent variables
Hyperbolic paraboloid 3D graph of overall acceptability was with flour formulations of 31.19 g of chickpea and 19.11 g of kidney
observed as shown in Figure 1c. The increase in the germinated bean with optimum responses. Predicted responses were found close
legume substitution actually lowered the overall acceptability of cook- and actual responses, which were obtained as 6.41 for spread ratio
ies, but overall acceptability within range for the product acceptability 41.98 N for snap force and overall acceptability of 4- on 5-point scale.
could be formed by interacting the two flours near to the saddle
points. The orthogonal function of equation provided the minimax
point in between two slopes, relative to both proportions. Axial points 3.4 | Characterization of optimized composite flour
delivered the high acceptance values, whereas central point provides and cookies from optimized formulation
the saddle point owing to interaction of both independent variables.
Therefore, as a result, interaction of both variables gave a positive 3.4.1 | Functional and pasting properties of
correlation and positive effect on overall acceptability. ungerminated wheat flour and OCFF

Functional properties like water absorption capacity, oil absorption

3.3 | Optimization of responses capacity, sedimentation value, foaming capacity, and emulsification
capacity were observed as reported in Table 3. Germination resulted
Model equations were reduced to evaluate the effect of variabilities in the increased protein content of flours; however, the resultant pro-
of responses with respect to the independent variables. Maximization tein content due to legume flour also attributed to variation in the
of desirable responses was required to optimize the final product for- functional and pasting properties. These properties are a type of bio-
mulations. Within range, independent variables were selected by the physical properties, which reveals the interaction of molecules with
design expert software to formulate the product with optimum the ingredients and thus provides the formal idea of flour behavior for
responses and a novel product formulation. Design expert provided a product formulation. Legume-blended flour (optimized composite
the valid and within range set of solutions. Selected solutions were flour formulation [OCFF]) exhibited higher water absorption capacity
analyzed to obtain the actual responses in the available set of condi- (1.46 ± 0.04) than did ungerminated wheat flour (UWF; 1.33 ± 0.02).
tions and then were compared with predicted values for validation Germination enhanced the disruption of polysaccharides that led to
SIBIAN AND RIAR 7

TABLE 3 Functional and pasting properties of optimized flour formulation and ungerminated wheat flour

Optimized composite flour formulation

Ungerminated wheat flour (UWF) (OCFF)
Functional properties
Water absorption capacity (g/g water 1.33 ± 0.02b 1.46 ± 0.04a
absorbed)
Oil absorption capacity (g/g oil 1.16 ± 0.02b 1.38 ± 0.03a
absorbed)
Sedimentation value (ml) 60.01 ± 0.01a 28.04 ± 0.02b
Foaming capacity (%) 12.76 ± 0.01b 21.43 ± 0.03a
Emulsification capacity (ml oil/g 18.56 ± 0.03b 24.45 ± 0.04a
sample)
Pasting properties
Peak viscosity (cP) 928.00 ± 0.34a 519.00 ± 0.47b
Trough viscosity (cP) 861.00 ± 0.41a 495.00 ± 0.38b
Break down (cP) 67.00 ± 0.51a 24.00 ± 0.43b
Final viscosity (cP) 1,124.00 ± 0.43a 934.00 ± 0.44b
Set back (cP) 263.00 ± 0.38b 439.00 ± 0.52a


Pasting temperature ( C) 88.20 ± 0.12a 83.85 ± 0.17b
Peak time (min) 6.87 ± 0.09a 5.67 ± 0.12b

Note: n = 3. Results are expressed as mean values ± standard deviations. Means in a column with different subscripts are significantly different (p < 0.05).

more damaged starch and thus retained more water. Another reason attainable viscosity at given temperature. Owing to degradation in
could be attributed to higher protein content in legumes, which starch, the trough viscosity of OCFF was reported to be lower
tended to absorb more water (Chauhan et al., 2015). Oil absorption (495.00 ± 0.38 cP). Breakdown determines the thermal and pasting
capacity of OCFF was also observed higher than in UWF. The stability of flour. Owing to germination and substitution of legumes
observed values for oil absorption capacity of OCFF were flour, the ability of pasting and thermal stability were higher in OCFF.
1.38 ± 0.03 g/g ,whereas UWF was reported to have 1.16 ± 0.02 g/g The observed values for breakdown in OCFF and UWF were
of oil absorbed. The increase in the protein content and hydrophobic 24.00 ± 0.43 and 67.00 ± 0.51 cP, respectively. Final viscosity is the
interaction of resultant protein molecules of blended flours could be index of flour's ability to form viscous paste. Final viscosity of
attributed to the variation in oil absorption capacities (Chiemela, ungerminated flour (1,124.00 ± 0.43 cP) was quite high as compared
Olufemi, & Joseph, 2009). Sedimentation value of raw wheat flour with that of blended flours (934.00 ± 0.44 cP). Higher retrogradation
was 60.01 ± 0.01 ml. With blending of different flours and germi- tendency and higher setback viscosity was shown by OCFF
nation, there was reduction in the total gluten fraction that led to (439.00 ± 0.52 cP). Lower retrogradation value leads to slow rate of
a lowered value of sedimentation. Germination and blending of staling process of product. UWF was observed with low setback vis-
flour affected the foaming capacity of composite flour. Foaming cosity (263.00 ± 0.38 cP). Pasting temperature correlates to gelatiniza-
capacity of OCFF (21.43 ± 0.03) was reported than that of UWF tion tendency of flour. Owing to degradation of starch, gelatinization
(12.76 ± 0.01). Foaming capacity is a function of surface properties tendency of legume-blended OCFF was lower. Peak time provides the
of some proteins (Sibian, Saxena, & Riar, 2017). Emulsification information for total time required for cooking of flour. Blending of
capacity of OCFF was also reported to be higher (24.45 ± 0.04). flours with different characteristics allowed the interaction of mole-
The variation in the emulsification capacities could be attributed to cules; therefore, the difference in cooking time was obtained as a
the protein–lipid interactions (Millward & Rivers, 1988; Sibian result (Adebowale et al., 2012).
et al., 2017).
Rheology of blended doughs was compared by using RVA, to
observe the pasting properties related to flours. RVA gave multi- 3.4.2 | Proximate analysis and in vitro digestibility of
parameter values like peak viscosity, trough viscosity, breakdown, cookies
final viscosity, setback viscosity, and pasting temperature to observe
the behavior of viscosity-related molecules (Table 3). Owing to germi- Table 4 indicates the difference in compositional characteristics of
nation, there was degradation of starch molecule, which leads to optimized composite flour cookies (OCFCs) and UWF cookies
lower value of peak viscosity in OCFF. Peak viscosity (cP) of UWF (UWFCs). Changes in the proximate composition of flours were previ-
was observed to be 928.00 ± 0.34, which was higher than that of ously reported in wheat and chickpea (Sibian et al., 2017; Sibian,
OCFF (519.00 ± 0.47). Trough viscosity is the measure of extreme Saxena, & Riar, 2016).
8 SIBIAN AND RIAR

TABLE 4 Compositional analysis and in vitro digestibility of TABLE 5 Texture profile analysis (TPA) and color characteristics
cookies from optimized composite formulation and ungerminated of cookies from optimized flour formulation and ungerminated wheat
wheat
Texture profile analysis
Ungerminated wheat Optimized composite Hardness Fracturability Cohesiveness
flour cookies (UWFCs) four cookies (OCFCs) (N) (N)
Moisture 3.57 ± 0.01b 4.89 ± 0.01a Ungerminated wheat 62.71 7.134 ± 0.03a 0.724
Protein 7.84 ± 0.02b 12.32 ± 0.11a flour cookies ± 0.05a ± 0.04a
(UWFCs)
Carbohydrates 69.75 ± 0.47a 54.43 ± 0.42b
Optimized composite 41.98 5.785 ± 0.03b 0.587
Fats 18.34 ± 0.26b 22.57 ± 0.23a
flour cookies ± 0.05b ± 0.01b
Crude fiber 1.93 ± 0.03b 5.64 ± 0.02a (OCFCs)
Ash 0.82 ± 0.02b 1.43 ± 0.02a Color characteristics
In vitro 38.67 ± 0.03b 50.27 ± 0.06a L* a* b*
carbohydrate
Ungerminated wheat 67.54 5.82 ± 0.03b 25.67
digestibility
flour cookies ± 0.07a ± 0.04a
In vitro protein 51.54 ± 0.05b 67.53 ± 0.04a (UWFCs)
digestibility
Optimized composite 52.37 9.13 ± 0.04a 30.84
Note: n = 3. Results are expressed in percentage and as mean values ± stan- flour cookies ± 0.04b ± 0.05a
dard deviations. Means in a column with different subscripts are signifi- (OCFCs)
cantly different (p < 0.05).
Note: n = 3. Results are expressed as mean values ± standard deviations.
Means in respective rows with different subscripts are significantly differ-
Moisture content was higher in OCFCs (4.89 ± 0.01) owing to ent (p < 0.05). L scale, lightness; a scale, red (+) vs. green (−); b scale, yellow
(+) vs. blue (−).
higher protein content of composite flour. With the substitution of
germinated flour of grains, the protein content of OCFCs
(12.32 ± 0.11) improved significantly. OCFCs tended to retain more hardness of cookies (Chauhan, Saxena, & Singh, 2016). Drakos,
oil during baking than did UWFCs (18.34 ± 0.26), which resulted in Andrioti-Petropoulou, Evageliou, and Mandala (2019) observed that
higher fat content in OCFCs (22.57 ± 0.23). Elin and Paul (2004) biscuits prepared from composite flour have softer and darker
observed the changes during substitution of wheat flour biscuits with appearance than has the control sample. Sindhuja et al. (2005)
germinated chickpea as compared with control sample containing observed a similar pattern of hardness in composite flour cookies,
10% chickpea control. Ash content was also higher in OCFCs by replacing wheat with 25–30% amaranth flour. Similar factors
(1.43 ± 0.02), whereas lower values were reported in UWFCs that influenced the hardness of cookies also affected the
(0.82 ± 0.02). Crude fiber content of OCFCs was quite higher and dif- fracturability and cohesiveness of cookies. Fracturability of cookies
fered significantly with that of the cookies prepared from UWF. Ger- varied in the range of 7.134 ± 0.03 and 5.785 ± 0.03, with the
mination increases the crude fiber content of grains owing to cell wall highest value observed in UWFCs. A higher cohesion value of
polysaccharides, and therefore, more fiber content was observed in UWFCs (0.724 ± 0.04) corresponded to their higher break strength,
OCFCs (5.64 ± 0.02). in vitro digestibility studies had shown the lower which could be affected by loss of intermolecular attractions
protein (51.54 ± 0.05) and carbohydrate digestibility (38.67 ± 0.03) in lu & Kotancilar, 2009). High protein and moisture content
(Karaog
UWFCs. The higher in vitro protein (67.53 ± 0.04) and carbohydrate could be the factors responsible for the lower cohesion
digestibility (50.27 ± 0.06) of OCFCs could be attributed to the pres- (0.587 ± 0.01) in OCFCs.
ence of free amino acid and breakdown of carbohydrate by the action Composite flour cookies were slightly darker than wheat flour
of enzyme alpha amylase during germination. Alonso, Aguirre, and cookies (Table 5). Color is one of the quality characteristics, which
Marzo (2000) conveyed that the in vitro protein digestibility of the reflects the cookie acceptability by the consumer. Color characteris-
Phaseolus vulgaris augmented to approximately 10% as a result of tics reflect the starch dextrinization, caramelization, and Maillard reac-
germination. tion, which are induced by cooking of product (Chung, Cho, &
Lim, 2014). Lightness of OCFCs was observed lower as compared
with that of UWFCs, which ranged from 52.37 ± 0.04 to
3.4.3 | Texture profile analysis and color 67.54 ± 0.07. The observations are in accordance with the results
characteristics obtained by Bolarinwa, Lim, and Muhammad (2018) in gluten-free
cookies prepared from germinated brown rice. Significant difference
Texture profile of cookies was analyzed for the components like was observed in the redness and yellowness of composite flour cook-
hardness, fracturability, and cohesiveness (Table 5). Hardness of ies and UWFCs. Maillard reaction could be accountable for the
UWFCs was reported 62.71 ± 0.05, which was higher than that of increased values of a* and b* (Islam, Taneya, Shams-Ud-Din,
OCFCs. A low level of gluten content due to substitution of Syduzzaman, & Hoque, 2012) in OCFCs owing to a higher amount of
legume flour caused the arrested gluten matrix, which affected the free amino acids and sugars.
SIBIAN AND RIAR 9

3.4.4 | Amino acid profile slightly during storage. With the increase in the moisture content,
other parameters like peroxide value, free fatty acid, and total plate
A higher amino acid content was observed in cookies prepared from count increased to some extent. The increase in the moisture content
an optimized formulation than in UWFCs. Essential amino acids could be attributed to the hygroscopic nature of cookies. The reason
(UWFCs/OCFC-g/100 g protein) like arginine (2.58 ± 0.01/ for legumes blended cookies to absorb high moisture could also be
2.66 ± 0.02), histidine (0.92 ± 0.03/0.95 ± 0.02), leucine (3.41 ± 0.03/ attributed to higher content of protein and crude fibers. Nagi, Kaur,
5.60 ± 0.04), lysine (0.97 ± 0.04/2.89 ± 0.03), methionine Dar, and Sharma (2012) reported similar findings in cereal bran-forti-
(0.90 ± 0.02/0.95 ± 0.04), phenylalanine (2.06 ± 0.02/2.32 ± 0.06), fied cookies at the end of 90 days. Peroxide value indicates the initial
threonine (1.88 ± 0.03/1.96 ± 0.03), tryptophan (0.57 ± 0.03/ rancidity and degree to which lipid undergoes primary oxidation.
0.70 ± 0.01), and valine (2.98 ± 0.02/3.22 ± 0.01) were reported sig- Owing to higher moisture and fat content of OCFCs, the peroxide
nificantly higher in OCFCs, whereas isoleucine (2.52 ± 0.02/ value was also reported to be higher. Cereals contain a lower amount
2.50 ± 0.06) was slightly higher in UWFCs. Legumes are generally of unsaturated fats; therefore, a lower amount of oxidation took place
deficient in methionine and correspondingly cereal lacks lysine, but in UWFCs (2.12 ± 0.01 to 5.16 ± 0.03). Peroxide value of OCFCs was
with the processing (germination and substitution) of flour, the formu- higher even during the initial day of study (4.12 ± 0.04), which
lated cookies were observed with good methionine and improved increased to 8.53 ± 0.03 mEq/kg on the 90th day. The results corrob-
lysine content. The reason for the good nutritional attribute of orated the studies by Divyashree (2014) in buckwheat–chia seed-for-
legume-blended cookies could be attributed to the amino acid profile tified cookies, which occurred owing to auto oxidation during storage.
of legume flours (Imran et al., 2011). Table 6 represents the total Ease of oxidation depends on the amount of unsaturated fats, storage
amino acid profile of UWFCs and OCFCs. Total aromatic amino acid conditions, and antioxidant activity of food (Akhter, Haider, Muzamil,
content (%) was found lower in OCFCs (8.27 ± 0.04) as compared Zia, & Salahuddin, 2016). Free fatty acid content was higher in OCFCs
with UWFCs (8.52 ± 0.06). Aromatic amino acid is more readily avail- at initial day (0.41 ± 0.03) owing to its lipid and moisture content. Dur-
able for utilization in chemical changes during baking (Oupadissakoon ing the storage, the increase in free fatty acid content of both OCFCs
& Young, 1984). Composite flour thus proved to be beneficial, owing and UWFCs was observed. The values of free fatty acid (mg KOH/g)
to the supplementation of deficient essential amino acids (Ikumola, at the 90th day for UWFCs and OCFCs were observed as 0.68 ± 0.05
Otutu, & Oluniran, 2017). and 0.89 ± 0.02, respectively. A similar trend of free fatty acids was
also reported in soy-fortified cookies (Singh, Singh, & Chauhan, 2000)
and composite pasta (Yadav, Sharma, Chikara, Anand, & Bansal, 2014)
3.4.5 | Storage studies with the increase in the storage time. Availability of moisture and
other nutritive components promote the growth of microbes in cook-
Multiple factors like moisture content, peroxide value, free fatty acids, ies. A higher moisture content was observed in OCFCs, which
and overall sensory score were observed for the storage studies of increased during storage. In addition, composite flour cookies were
cookies (Table 7). Moisture content of cookies sample increased reported to be high in nutrients, which facilitated the microbial
growth. Germination converts the complex molecules into simpler
TABLE 6 Amino acid profile of cookies from optimized molecules like sugar and amino acids, which might be readily available
composite flour formulation and ungerminated wheat to the microorganism. Therefore, as a result, the microbial load of

Ungerminated wheat Optimized composite cookies increased. According to Gilbert (2000), the permissible limit
flour cookies (UWFCs) flour cookies (OCFCs) for total aerobic count of ready-to-eat foods should be less than 104–
Total 53.31 ± 0.04a 36.88 ± 0.03b 106 cfu/g. The microbial count of UWFCs was reported in the range
nonessential of 1.5 × 101 to 1.03 × 102 cfu/g during the 90-day storage period.
amino acid The initial microbial count in OCFCs was 2.0 × 101 and increased
Total essential 46.69 ± 0.06b 63.12 ± 0.05a up to 1.07 × 102 on the 90th day of storage. Although there was
amino acid an increment in microbial count, the total plate count during the
Total aromatic 8.52 ± 0.06a 8.27 ± 0.04b storage period of preservation did not exceed the permissible limit.
amino acid
Yusufu, Netala, and Opega (2016) also observed an increase in the
Total basic 11.07 ± 0.03b 20.32 ± 0.04a
total viable count (cfu/g) in composite flour cookies prepared from
amino acid
maize, yam bean, and plantain during storage. Organoleptic proper-
Total acidic 30.42 ± 0.07a 19.62 ± 0.05b
ties depend on various intrinsic and extrinsic factors (Mudgil, Barak,
amino acid
& Khatkar, 2017). Sensory evaluation showed that the optimized
Sulfur 3.86 ± 0.03ab 3.85 ± 0.05ba
containing formulated cookies were acceptable even after the storage period
amino acid of 90 days. Results for legume-substituted cookies were in agree-
ment with the studies of Noor Aziah, Mohamad Noor, and
Note: n = 3. Results are expressed in percentage and as mean values ± stan-
dard deviations. Means in columns with different subscripts are signifi- Ho (2012) on the organoleptic properties of cookies fortified with
cantly different (p < 0.05). legume.
10 SIBIAN AND RIAR

TABLE 7 Partial shelf life studies of cookies from optimized formulation and ungerminated wheat

Number of Peroxide values Free fatty acid content

days Moisture content (mEq/kg) (mg KOH/g) Microbial count (cfu/g) Overall sensory score

UWFCs OCFCs UWFCs OCFCs UWFCs OCFCs UWFCs OCFCs UWFCs OCFCs
0 3.86 5.12 2.12 4.12 0.29 0.41 1.5 × 10 1
2.0 × 101
4.10 4.00
± 0.02 ± 0.01 ± 0.01 ± 0.04 ± 0.01 ± 0.03 ± 0.07 ± 0.08
10 3.98 5.42 2.33 4.49 0.34 0.49 1.8 × 101 2.4 × 101 4.07 4.00
± 0.01 ± 0.01 ± 0.02 ± 0.03 ± 0.01 ± 0.03 ± 0.06 ± 0.09
20 4.25 5.66 2.67 4.63 0.41 0.57 2.5 × 101 2.9 × 101 4.04 3.99
± 0.02 ± 0.01 ± 0.02 ± 0.02 ± 0.02 ± 0.02 ± 0.08 ± 0.09
30 4.34 5.83 2.92 5.07 0.43 0.62 2.9 × 101 3.3 × 101 4.02 3.97
± 0.03 ± 0.01 ± 0.05 ± 0.04 ± 0.04 ± 0.04 ± 0.08 ± 0.07
40 4.69 5.63 3.18 5.42 0.47 0.71 3.2 × 101 4.9 × 101 4.00 3.96
± 0.02 ± 0.01 ± 0.02 ± 0.03 ± 0.06 ± 0.04 ± 0.06 ± 0.08
50 4.77 5.83 3.26 5.76 0.49 0.74 4.1 × 101 5.3 × 101 3.98 3.93
± 0.04 ± 0.01 ± 0.03 ± 0.03 ± 0.03 ± 0.05 ± 0.09 ± 0.07
60 4.89 6.11 3.43 6.38 0.54 0.76 5.5 × 101 6.9 × 101 3.95 3.91
± 0.03 ± 0.01 ± 0.02 ± 0.06 ± 0.02 ± 0.02 ± 0.08 ± 0.09
70 5.03 6.27 4.31 6.92 0.59 0.81 7.1 × 101 7.7 × 101 3.92 3.88
± 0.03 ± 0.01 ± 0.04 ± 0.05 ± 0.04 ± 0.02 ± 0.09 ± 0.05
80 5.14 6.39 4.94 7.34 0.64 0.84 9.7 × 101 9.8 × 101 3.90 3.85
± 0.02 ± 0.01 ± 0.03 ± 0.03 ± 0.02 ± 0.04 ± 0.09 ± 0.11
90 5.19 6.48 5.16 8.53 0.68 0.89 1.03 × 102 1.07 × 102 3.87 3.82
± 0.02 ± 0.01 ± 0.03 ± 0.03 ± 0.05 ± 0.02 ± 0.07 ± 0.07

Note: n = 3. Results are expressed as mean values ± standard deviations. Values in bold are significantly different from other values in respective columns
for different parameters (p < 0.05).
Abbreviations: OCFCs, optimized composite flour cookies; UWFCs, ungerminated wheat flour cookies.

4 | CO NC LUSIO N CONFLIC T OF INT ER E ST

The authors declare no conflict of interest with respect to this
The two-way approach of formulating a novel product for the prepa- manuscript.
ration of cookies delivered a nutritious option in place of traditionally
prepared cookies from ungerminated wheat. Various attempts have AUTHOR CONTRIBU TIONS
been made previously by researchers to successfully replace wheat Charanjit Singh Riar conceived and supervised the concept, and
flour with other legumes and cereals. Germinated cereal and legumes Mandeep Singh Sibian performed the experiments, analyzed the data,
are able to replace the traditionally prepared wheat cookies for better and wrote the manuscript.
digestibility and nutrition. Functional and pasting properties of
legume-blended flour have shown the suitability of substitution and ET HICS S TAT E MENT
utilization of different flours from germinated grains. Owing to alter- This article does not contain any human and animal subjects for
ation in pasting properties and functional characteristics, cookies pre- experiment.
pared from the composite flour exhibited soft texture cookies.
Textural properties of OCFCs were softer with slightly dark color. All DATA AVAILABILITY STAT EMEN T
age groups prefer these kinds of cookies with soft texture and high The data that support the findings of this study are available from the
digestibility. The shelf life of OCFCs was found quite comparable with corresponding author upon reasonable request.
that of UWFCs, whereas sensory score has shown the acceptability of
product on a 5-point scale. Shelf life could be extended to a longer
OR CID
period by using specialized packaging techniques.
Mandeep Singh Sibian https://orcid.org/0000-0002-1594-3226

ACKNOWLEDGEMEN TS
RE FE RE NCE S
The authors acknowledge the Department of Food Engineering and
Adebowale, A. A., Adegoke, M. T., Sanni, S. A., Adegunwa, M. O., &
Technology, Sant Longowal Institute of Engineering and Technology,
Fetuga, G. O. (2012). Functional properties and biscuit making poten-
Longowal, for providing necessary infrastructure for this research tials of sorghum-wheat flour composite. American Journal of Food
work. No funds were received from any agency for this work. Technology, 7(6), 372–379.
SIBIAN AND RIAR 11

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