Physicochemical, Pasting, and Functional

Properties of Amaranth Seed Flours: Effects

of Lipids Removal
Khetan Shevkani, Narpinder Singh, Amritpal Kaur, and Jai Chand Rana

C: Food Chemistry
Abstract: The present work was carried out to evaluate physicochemical (composition, hunter color, and sodium dodecyl
sulfate–polyacrylamide gel electrophoresis [SDS-PAGE]), pasting, and functional properties (foaming, emulsification,
water, and fat absorption capacity) of amaranth full-fat flours from 6 lines/cultivars (AFs), and to see the effects of lipid
removal/defatting on these properties. Protein, ash, and lipid content of AFs ranged between 12.5% to 15.2%, 3.0% to
3.5%, and 7.1% to 8.0%, respectively. The flours showed a number of bands between 97 and 7 kDa, with main subunits
of approximately 58, 37, 33, 31, 23, and 16 kDa in the SDS-PAGE profiles. The protein content and L∗ value increased,
while b∗ values decreased following defatting for most of the lines/cultivars. The defatted flours (DAFs) had higher final
viscosity and stability (lower breakdown viscosity) as compared to counterpart AFs. The protein profiling of the flours
was not affected with the lipid removal/defatting. However, water absorption capacity and foam stability of the flours
improved upon defatting. Principal component analysis revealed that pasting temperature was positively related to lipid
content, while breakdown viscosity was negatively related to protein content. Foaming properties (capacity and stability)
showed negative relationship with lipid content, and positive with protein content, ash content, water, and fat absorption
capacity.

Keywords: amaranth flour, functional, lipid removal/defatting, pasting, SDS-PAGE

Practical Application: Amaranth grains are known to have higher amount of proteins and lipids than cereals. Amaranth
lipids are rich in unsaturated fatty acids, which are prone to oxidative rancidity. Removal of lipids or defatting of flours
may be carried out to enhance product shelf life by preventing undesirable oxidative chain reactions. Therefore, this
research was undertaken to see the effects of defatting on the functional properties of amaranth flours. The defatting was
a value addition process as it improved the functional properties of the flours.

Introduction
Amaranth is a dicotyledonous plant that produces large quan-
tities of edible grains and presents many agronomic advantages
such as relative high grain yield, resistance to drought, and short
production time (Mendoza and Bressani 1987). Amaranth has re-
cently become a focus of interest for its high nutritive values and
cholesterol-lowering effect observed in animal models (Plate and
Areas 2002; Mendonca and others 2009).
Amaranth have higher protein content (12% to 18%) than most
of the cereals with a significant higher content of lysine and accept-
able level of tryptophan and methionine, which are found in low
concentration in cereal and legume grains (Mendoza and Bressani
1987). Amaranth grains contain 48% to 69% (dry basis) starch,
which is located in perisperm as very small granules of 0.75 to 3.0
μm in diameter (Hoover and others 1998). The small granular
size of the amaranth starch makes it suitable for different food and
nonfood applications such as fat replacers, food thickeners, paper
coatings, dusting powders, laundry starch, and biodegradable plas-
tics (Hoover and others 1998). In addition to protein and starch,
MS 20131740 Submitted 11/22/2013, Accepted 4/12/2014. Authors Shevkani,
Singh, and Kaur are with Dept. of Food Science and Technology, Guru Nanak
Dev Univ., Amritsar-143005, India. Author Rana is with Natl. Bureau of Plant
Genetics Resources, Shimla-171004, India. Direct inquiries to author Singh (E-mail:
narpinders@yahoo.com).
the amaranth is a good source of lipids, dietary fiber, and minerals
(magnesium, phosphorus, copper, manganese, and so on). The
amaranth is also a rich source of polyphenols (flavonoids) with
relatively high antioxidant activity. Caffeic acid, p-hydroxybenzoic
acid, and ferulic acid are the main phenolic compounds in ama-
ranth grains (Singh and Singh 2011). Amaranth grains may contain
up to 10% oil that is similar to other cereals consisting mainly of
unsaturated fatty acids, with linoleic acid being the predominant
fatty acid. Amaranth lipids also contain squalene (Sun and others
1997), which has anticarcinogenic and hypocholesterolemic effects
(Shin and others 2004; Escudero and others 2006). The amaranth
oil thus has the potential to be used for several nutraceutical appli-
cations and replace commercial squalene sources such as shark and
whale.
The amaranth grains are consumed in roasted and sprouted
form, while flour is used as thickener in gravies, soups, and stews.
Amaranth flour may also find application in preparation of bis-
cuits, muffins, pancakes, pastas, flat breads, and so on, because of
its nutritional properties (Singh and Singh 2011). The utilization
of amaranth flour in food system greatly depends on its func-
tional properties. Hydration, fat absorption, emulsification, foam-
ing, and viscosity are important functional properties of flours that
can decide their utilization for different foods applications (meat,
confectionary, and bakery products).
The removal of lipids from the flours may have beneficial effects,
for example, prevention of oxidative reactions; extracted oil can be

C 2014 Institute of Food Technologists

 R

doi: 10.1111/1750-3841.12493 Vol. 79, Nr. 7, 2014 r Journal of Food Science C1271
Further reproduction without permission is prohibited
 Functional properties of amaranth flours . . .
used for other purposes; defatted flours (DAFs) can be marketed
as low-fat or low-calorie products; and flour properties may be
modified, and so on. Recently, attention is being given to study
the effect of defatting on the functional properties of different
flours and proteins (L’Hocine and others 2006; Fasasi and oth-
ers 2007; Jitngarmkusol and others 2008; Alobo and others 2009;
Ogunsina and others 2010; Toews and Wang 2013). Toews and
C: Food Chemistry
Wang (2013) reported that functional properties of the chickpea
protein concentrates were improved significantly when a defatting
step was added prior to the fractionation process. Fasasi and others
(2007) and Jitngarmkusol and others (2008) reported that defat-
ting significantly influenced the functional properties of breadfruit
seed and macadamia flours, respectively. Jitngarmkusol and others
(2008) showed that DAFs had enhanced nutritional and functional
properties than full-fat flours.
Tapia-Blacido and others (2010) characterized Amaranthus cru-
entus BRS Alegria flour, starch, and protein concentrates and
concluded that they have interesting characteristics for use as
food ingredients. Kaur and others (2010) studied diversity in the
plant, grains, and flour characteristics among different amaranth
lines. The physicochemical and functional properties of amaranth
proteins, starch, and fiber have been discussed in the literature
(Gorinstein and others 1991; Barba de la Rosa and others 1992;
Repo-Carrasco-Valencia and others 2009; Shevkani and others
2014a; Shevkani and Singh 2014; Singh and others 2014a). How-
ever, studies on the physicochemical and functional properties of
full-fat and DAFs are not available. Therefore, the present research
was undertaken to study physicochemical, pasting, and functional
properties of amaranth flours, and to investigate the effects of lipid
removal/defatting on these properties.
Materials and Methods
Materials
Amaranth seeds (Amaranthus hypochondriacus) of 6 lines/cultivars,
IC 35407, PRA 01, PRA 03, Annpurna, IC 540862, and IC
95341, were grown in the year of 2008 at Regional Centre, Natl.
Bureau of Plant Genetic Resources, Shimla, India. The grains
were cleaned and ground in a Super mill (Newport Scientific,
Australia) to pass all through a sieve with 250 μm aperture. The
flours were defatted thrice using hexane at a flour/hexane ratio
of 1 : 4 for 8 h (total defatting time 24 h). After each extraction,
the solvent was separated by filtering the slurry through Whatman
filter paper (nr. 1). Finally, the defatted residue was dried in a
laboratory oven at 35 °C for 24 h. The defatting was done for at
least 2 different batches of the flour for each lines/cultivars, those
were then mixed together. The amaranth full-fat flours (AFs) and
respective DAFs from different lines/cultivars were stored at 4 °C
in air tight PET jars till analysis.
Proximate composition
Samples were estimated for their moisture, ash, fat, and protein
(N × 5.85) content by employing the standard methods (AOAC
1990). Total carbohydrate content was calculated as difference.
Hunter color parameters
Color of the AFs and DAFs was measured using Ultra Scan
VIS Hunter Lab (Hunter Associates Laboratory Inc., Reston Va.,
U.S.A.). Color parameters were expressed as L∗ , a∗ , and b∗ values
with L∗ being for lightness, a∗ for redness–greenness, and b∗ for
yellowness–blueness of the flours.
C1272 Journal of Food Science r Vol. 79, Nr. 7, 2014
Pasting properties
Pasting properties of AFs and DAFs from different lines/cultivars
(10% suspensions) were determined using a rheometer (AntonPaar,
Physica MCR 301, Ostfildern, Germany) equipped with starch
cell (C-ETD 160) and stirrer probe (ST 24-2D/2V/2V-30). The
temperature–time conditions included a heating phase from 50 to
95 °C at the rate of 6 °C/min and a holding phase at 95 °C for
5 min, a cooling phase from 95 to 50 °C at a rate of 6 °C/min,
and a holding phase at 50 °C for 2 min. Pasting temperature (PT),
peak viscosity (PV), breakdown viscosity (BV), and final viscosity
(FV) were obtained from the pasting profile of flours.
Functional properties
Foaming properties including foaming capacity (FC) and foam
stability (FS), emulsifying properties (Pearce and Kinsella 1978),
that is, emulsifying activity index (EAI) and emulsifying stability
index (ESI), water absorption capacity (WAC), and fat absorp-
tion capacity (FAC) of AFs and DAFs from different lines were
determined as described elsewhere (Shevkani and others 2014a).
Sodium dodecyl sulfate–polyacrylamide gel
electrophoresis (SDS-PAGE)
Total proteins from AFs and DAFs were extracted following
the method of Drzewiecki and others (2003). Briefly, 40 mg
flour samples were stirred in 1 mL tris buffer pH 6.8, containing
sodium dodecyl sulfate, β-mercaptoethanol, bromophenol blue,
and glycerol. The samples were boiled for 5 min and then cen-
trifuged at 13600 × g for 10 min at room temperature. SDS-
PAGE was performed according to Laemmli (1970) using 13%
polyacrylamide resolving gel and 5% polyacrylamide stacking gel.
The proteins were resolved at a constant current of 35 mA. The
gel was stained overnight using Coomassaie Brilliant Blue R 250
(0.25%) and destained using 20% methanol and 10% acetic acid
solution.
Statistical analysis
The reported data are mean of triplicate observations. The data
were subjected to analysis of variance by Duncan’s test (P < 0.05)
and principal component analysis (PCA) using Minitab Statistical
Software (State College, Pa., U.S.A.). PCA was carried out in
order to determine relationships between physicochemical and
functional properties among different flours as well as to determine
the similarities/differences among different lines/cultivars on the
basis of these properties.
Results and Discussion
Proximate composition and color properties
The proximate composition of AFs and DAFs is presented
in Table 1. The results indicated that the defatting significantly
increased the protein and carbohydrate contents of the flours from
most of the lines/cultivars. The protein and carbohydrate content
of the AFs ranged between 12.5% to 15.2% and 73.7% to 77.0%
(db), respectively, that increased to 13.2% to 16.3% and 79.2% to
82.5% (db), respectively, after defatting (Table 1). The ash content
of the AFs ranged between 3.0% and 3.5% (db). The defatting of
AFs from different lines/cultivars except PRA 03 did not cause
a significant increase in ash content (Table 1). The lipid content
of the AFs varied between 7.1% and 8.0% (db). The defatting
of AFs removed most of the lipids; however, a part of it (0.4%
to 1.1%) remained in the flours. These residual lipids might be
Functional properties of amaranth flours . . .

Table 1–Proximate composition (dry weight basis) of full-fat and defatted amaranth flours from different lines/cultivars.

Protein content (%) Ash content (%) Lipid content (%) # Carbohydrate content
Line/cultivar AF DAF AF DAF AF DAF AF DAF
IC 35407 15.2 ± 0.2Ac 16.3 ± 0.3Bc 3.1 ± 0.2Aa 3.4 ± 0.0Aab 8.0 ± 0.0Bc 1.14 ± 0.2Ab 73.7 ± 0.0Aa 79.2 ± 0.6Ba
PRA 01 12.5 ± 0.1Aa 13.2 ± 0.2Ba 3.0 ± 0.1Aa 3.3 ± 0.1Aa 7.5 ± 0.1Bab 0.97 ± 0.1Ab 77.0 ± 0.1Ac 82.5 ± 0.2Bc
PRA 03 13.1 ± 0.6Aab 14.6 ± 0.1Bb 3.5 ± 0.1Ab 3.8 ± 0.0Bb 7.1 ± 0.1Ba 0.39 ± 0.1Aa 76.4 ± 0.7Ac 81.2 ± 0.0Bb

C: Food Chemistry
Annpurna 13.9 ± 0.1Ab 14.5 ± 0.4Bb 3.2 ± 0.1Aa 3.2 ± 0.2Aa 7.5 ± 0.3Bab 1.13 ± 0.1Ab 75.4 ± 0.2Ab 81.1 ± 0.2Bb
IC 540862 13.1 ± 0.1Aab 13.4 ± 0.1Aa 3.1 ± 0.1Aa 3.2 ± 0.3Aa 7.6 ± 0.1Bab 1.03 ± 0.1Ab 76.2 ± 0.1Abc 82.3 ± 0.4Bc
IC 95341 14.0 ± 0.4Ab 15.7 ± 0.3Bc 3.2 ± 0.3Aa 3.4 ± 0.0Aab 7.2 ± 0.1Ba 0.98 ± 0.1Ab 75.5 ± 0.2Ab 79.9 ± 0.3Ba
Values are means ± SD. Means with similar lower case letters shown as superscript in a column did not differ significantly (P > 0.05). Means with similar capital letter in a row for
each parameter did not differ significantly (P > 0.05). AF, amaranth full fat flour; DAF, defatted amaranth flour. # calculated as difference.

Table 2–Hunter color properties of full-fat and defatted amaranth flours from different lines/cultivars.

L∗ a∗ b∗
Line/cultivar AF DAF AF DAF AF DAF
IC 35407 82.31 ± 0.06Aa 83.35 ± 0.10Ba 2.23 ± 0.04Bd 1.82 ± 0.03Ab 13.35 ± 0.10Bc 11.01 ± 0.17Aa
PRA 01 85.40 ± 0.16Ab 86.72 ± 0.11Bc 1.45 ± 0.03Bb 1.29 ± 0.01Aa 13.14 ± 0.14Bbc 11.06 ± 0.09Aa
PRA 03 85.77 ± 0.06Ab 86.17 ± 0.11Bbc 1.31 ± 0.01Aa 1.34 ± 0.02Aa 12.63 ± 0.10Ba 10.77 ± 0.18Aa
Annpurna 85.59 ± 0.09Ab 86.34 ± 0.27Bc 1.42 ± 0.03Aab 1.37 ± 0.06Aa 13.02 ± 0.21Bb 11.01 ± 0.14Aa
IC 540862 85.25 ± 0.22Ab 85.63 ± 0.18Ab 1.64 ± 0.01Bc 1.37 ± 0.10Aa 12.65 ± 0.04Aa 11.97 ± 0.28Ab
IC 95341 85.10 ± 0.18Ab 85.49 ± 0.34Ab 1.40 ± 0.02Aab 1.38 ± 0.01Aa 12.67 ± 0.05Ba 10.90 ± 0.27Aa
Values are means ± SD. Means with similar lower case letters shown as superscript in a column did not differ significantly (P > 0.05). Means with similar capital letter in a row for
each parameter did not differ significantly (P > 0.05). AF, amaranth full fat flour; DAF, defatted amaranth flour.

starch-bound lipids that could not be removed using hexane.
Hoover and others (1998) reported 0.16% to 0.28% starch-bound
lipids for starches from 3 amaranth cultivars. Additionally, the
presence of polar lipids interacted with proteins cannot be ruled
out (Kikugawa and others 1981). Proteins and lipids significantly
influence the physicochemical and pasting properties of flours
(Shevkani and others 2014b). The ash, lipid, and protein content
of the AFs were within the range (1.7% to 3.8%, 3.8% to 7.4%,
and 6.6% to 19.8%, respectively) observed earlier for 48 A.
hypochondriacus lines (Kaur and others 2010) and were comparable
to those reported for A. caudatus (2.14%, 8.93%, and 14.21%,
respectively) and A. cruentus BRS Alegria flours (2.10%, 7.96%,
and 14.08%, respectively) (Tapia-Blacido and others 2010). Jit-
ngarmkusol and others (2008) reported that defatted macadamia
flours had higher protein, ash, and carbohydrate content than
counterpart partially DAFs.
Color is one of the most important characteristics that can de-
cide successful applications of functional ingredients in different
food products. L∗ , a∗ , and b∗ values of AFs ranged between 82.31
to 85.77, 1.31 to 2.23, and 12.63 to 13.35, respectively. All flours
showed positive a∗ values, indicating a red tint in AFs. The b∗
value, an indicator of blue (−) and yellow (+), indicated the pres-
ence of yellowness in all the flours. Defatting of the AFs influenced
PCA analysis revealed that the DAFs were different than those
of AFs in terms of the measured properties as AFs were located
at right of the plot, while those of DAFs were located on the left
pane of the score plot (Figure 1). The PCA score plot provides
ready means of visualizing the differences or similarities among
different flours in terms of the measured properties. The distance
between the locations of any 2 line/cultivar on the score plot
is directly proportional to the degree of difference or similarity
between them (Kaur and others 2007).
Pasting properties
The pasting curve represents changes in behavior of flour paste
viscosity with change in temperature and mainly varies with flour
composition and characteristics of starch. During heating, the vis-
cosity of flour suspension increased due to the swelling of the
granules to several times their original size, as a result of loss of
crystalline order and absorption of water (Bao and Bergman 2004).
The temperature at which the viscosity of flour suspension begins
4
IC 35407 DF
Second Component

IC 35407
their color parameters (Table 2). However, the effect of defatting 2 IC 54862 DF
on the color was observed to be dependent on line/cultivar. DAFs Annpurna DF IC 54862
from IC 35407, PRA 01, PRA 03, and Annpurna had significantly PRA 03 DF Annpurna
0
higher L∗ value (lightness) as compared to their counterpart AFs. PRA 03
IC 95341 DF
The lines/cultivars (except IC 540862) had significantly lower yel- IC 95341
lowness than the counterpart AFs (Table 2). The increase in L∗ -2
PRA 01
and decrease in b∗ values may be due to the removal of oil-soluble
PRA 01 DF
coloring pigments. Higher L∗ value (lightness) of DAFs may be a
-4
preferred characteristic for their application as a functional ingre- -4 -2 0 2 4
dient in various products. The color of functional additives should First Component
have minimal detrimental effects on the food product (Toews and
Wang 2013). Higher L∗ value while lower a∗ and b∗ values for Figure 1–Principal component analysis score plot describing overall simi-
protein concentrates from defatted chickpeas than their undefatted larities or differences amongst full-fat and defatted amaranth flours from
counterparts have been reported earlier (Toews and Wang 2013). different lines/cultivars.

Vol. 79, Nr. 7, 2014 r Journal of Food Science C1273

 Functional properties of amaranth flours . . .

Table 3–Pasting properties of full-fat and defatted amaranth flours from different lines/cultivars.

Pasting temperature (°C) Peak viscosity (cP) Breakdown viscosity (cP) Final viscosity (cP)
Line/cultivar AF DAF AF DAF AF DAF AF DAF
IC 35407 71.3 ± 0.16Aa 71.3 ± 0.45Aab 1121 ± 29Aab 1175 ± 21Bb 382 ± 21Aa 355 ± 8Ac 966 ± 18Aa 1032 ± 16Ba
PRA 01 71.9 ± 0.36Aa 71.7 ± 0.33Ab 1459 ± 13Ad 1543 ± 5Bd 475 ± 3Bb 418 ± 5Ad 1286 ± 8Ad 1354 ± 4Be
PRA 03 72.1 ± 0.17Ba 70.7 ± 0.18Aa 1200 ± 33Ab 1184 ± 27Ab 389 ± 19Ba 295 ± 5Ab 1068 ± 28Ab 1127 ± 14Bc
C: Food Chemistry

Annpurna 71.7 ± 0.05Ba 70.7 ± 0.35Aa 1213 ± 20Ab 1167 ± 22Ab 403 ± 4Ba 299 ± 11Ab 1056 ± 13Ab 1107 ± 13Bbc
IC 540862 71.7 ± 0.28Ba 70.5 ± 0.26Aa 1050 ± 9Aa 1068 ± 19Aa 398 ± 13Ba 237 ± 22Aa 995 ± 23Aab 1077 ± 5Bb
IC 95341 72.1 ± 0.47Ba 71.1 ± 0.16Aab 1352 ± 32Ac 1422 ± 34Bc 435 ± 17Bab 405 ± 8Ad 1193 ± 25Ac 1283 ± 21Bd
Values are means ± SD. Means with similar lower case letters shown as superscript in a column did not differ significantly (P > 0.05). Means with similar capital letter in a row for
each parameter did not differ significantly (P > 0.05). AF, amaranth full fat flour; DAF, defatted amaranth flour.

to rise was taken as PT. PT represents minimum temperature re-
quired for cooking. PT of AFs ranged between 71.3 and 72.1 °C
that reduced to 70.5 and 71.7 °C following defatting; however,
the decrease in the PT was statistically significant for PRA 03,
Annpurna, IC 540862, and IC 95341 only (Table 3). PV repre-
sents the point of maximum swelling of the starch granules, ranged
between 1050 and 1459 cP for AFs from different lines/cultivars.
Heating of gels at elevated temperatures resulted in a drop in the
viscosity attributing to the rupturing of starch granules (Figure 2).
BV, a measure of the resistance of gel to disintegrate at high tem-
peratures ranged between 282 and 475 cP. Lower BV represents
greater resistance to shear thinning and higher stability of flour
pastes. Upon cooling, the viscosity of cooked flour gels increased
as a result of retrogradation or tendency of gel components to
reassociate. The FV indicating the ability of the flours to form
a viscous paste, ranged between 966 and 1286 cP. The defatting
improved paste characteristics of flours, as DAFs showed higher
FV, and lower BV than counterpart AFs (Table 3). A decrease
in viscosity with increasing lipids content had been observed for
wheat, oat, and rice flour (Singh and Smith 1999; Dautant and
others 2007). The lubricating effect of lipids might be attributable
for lower viscosities of AFs (Dautant and others 2007). Addition-
ally, the effect of starch–lipid complexes formed during heating,
on the pasting properties cannot be ruled out. The lipids can form
complexes with starch during biosynthesis of the starch as well
as during heating of starch slurries at gelatinization temperature
and above (Singh and other 2014b) that can increase granular in-
tegrity, leading to higher PT and retardation of granular swelling
and lower paste viscosities (Shevkani and others 2011). The role of
lipids on pasting properties was also observed as positive relation
with PT, while negative relations with PV and FV on PCA load-
ing plot (Figure 3). The PCA loading plot provides an overview
of the interrelationships between different parameters. The pa-
125
rameters whose curves lie close to one another on the plot are
positively related, while those whose curves run in opposite di-
rections are negatively related. The defatting also improved paste
stability and resistance to shear thinning of flours, as DAFs showed
lower BV than counterpart AFs; however, the improvement was
statistically insignificant (P > 0.05) for IC 35407 (Table 3). Lower
BV following defatting might be due to enhanced protein con-
tent that protected starch granules against disintegration at high
temperature and shear, which was supported by a strong negative
relationship between BV and protein content on the loading plot
(Figure 3). The denatured proteins stabilize the continuous ma-
trix between the dispersed and continuous phases and result in
lower breakdown of flour pastes (Fitzgerald and others 2003). A
strong negative correlation between BV and protein content has
been reported for corn fractions obtained from successive reduc-
tion stages during dry milling (Shevkani and others 2014b). An
increase in peak and FV and decrease in BV after defatting had
been observed for oat meal (Zhou and others 1999). The results
clearly indicated that DAFs in the presence of lower lipids and
higher proteins had higher FV and improved paste stability during
heating and shearing.
Functional properties
The functional properties are the properties of flours that pri-
marily determine their utilization in different food products.
Foaming properties (capacity and stability) are important func-
tional characteristics of flours that determine their utilization in
food systems, where aeration and overrun is required, for example,
whipped toppings, baked foods, and ice cream. FC and FS of AFs
varied significantly between 15% to 30% and 17% to 50%, respec-
tively. The FC of AFs was, however, lower than that of different
0.50
a*
EAI
Second Component

1600 PC
0.25
DAF
100 FC
AC
AF
Temperature (°C)

1200 FS LC
0.00
Viscosity (cP)

WAC b*
75

800 FAC ESI

50 -0.25 PT
L* BV
400
25
FV PV
-0.50
-0.50 -0.25 0.00 0.25 0.50
0 0
0 5 10 15 20 25 First Component
Time (min)

Figure 3–Principal component analysis loading plot describing relationship

Figure 2–Representative pasting profile of full-fat (AF) and defatted (DAF) among different properties of full-fat and defatted amaranth flours from
amaranth flour (PRA 01). different lines/cultivars.

C1274 Journal of Food Science r Vol. 79, Nr. 7, 2014

Functional properties of amaranth flours . . .

Chinese legume flours (between 64.4% and 140% at different pH)
reported by Chau and Cheung (1998). The foaming properties
are dependent on the proteins as well as on other components
such as carbohydrates (Ma and others 2011); therefore, the vari-
ations in foaming properties among different lines/cultivars may
be attributed to the differences in their composition. DAFs from
most of the lines/cultivars showed higher FC (25% to 35%) and
and give stabilized foam. FC is related to the proteins’ ability to
rapidly diffuse to the interface, reorient, and form a viscous film
without excessive aggregation or coagulation, whereas foaming
stability is influenced by intermolecular cohesiveness and viscos-
ity of this interfacial film as well as a certain degree of elasticity
permitting localized contact deformation (Kinsella 1981). The
improvements in FS after defatting of the flours might be due to

FS (50% to 79%) than AFs (Figure 4). FC of 2 lines/cultivars
(PRA 03 and Annpurna) did not significantly improved on de-
fatting, whereas these lines/cultivars showed the highest FS after
removal of lipids, suggesting that DAFs of these lines/cultivars are
particularly suitable for products where dense and stable foams
are desirable. An increase in the FC from 22.67% to 33.67% to
62.33% to 126.0% for different macadamia flours (Jitngarmkusol
and others 2008) and from 4.2% to 8.4% for breadfruit seed flour
(Fasasi and others 2007) following defatting had been reported.
Foam is a colloid of many small air bubbles that are surrounded by
thin liquid film. The flours produce foam when whipped mainly
because of the surface active proteins that reduce surface ten-
sion at the air-liquid interface, avoid coalescence of the bubbles
C: Food Chemistry
partial denaturation of the proteins during defatting (Damodaran
2005) that might have increased viscosity, which, in turn, imparted
rigidity to the interfacial film for foam stabilization. Conforma-
tional changes in proteins induced by unfolding and subsequent
interactions can occur during defatting that can contribute to im-
provement in foaming properties (Kinsella 1981). Furthermore,
the lipids markedly impair foaming properties of flours as lipids
are more surface active than proteins and are readily adsorbed
at interface and inhibit adsorption of proteins during foaming
(Damodaran 1997). It has been reported that legume protein con-
centrates with high-fat content produced low and unstable foams
(Traina and Breene 1994; Toews and Wang 2013). The role of
proteins and lipids on foaming properties was also depicted on

45 90

30 60
FC (%)

FS (%)

15 30

0 0
7

1
a

a
01

03

01

03
40

86

34

40

86

34
rn

rn
u

u
A

A
35

54

95

35

54

95
np

np
PR

PR

PR

PR
IC

IC

IC

IC

IC

IC
An

An

45 140

105
EAI (m2/g)

ESI (min)

30
70

15
35

0 0
7

1
a
01

03
7

1
a
01

03

40

86

34
rn
40

86

34
rn

u
A

A
35

54

95
u
A

A
35

54

95

np
PR

PR
np
PR

PR

IC

IC

IC
An
IC

IC

IC
An

3 3
WAC (g/g)

FAC (g/g)

2 2

1 1

0 0
7

1
a

1
01

03

a
01

03
40

86

34
rn

40

86

34
rn
u
A

u
35

54

95

A
35

54

95
np

np
PR

PR

PR

PR
IC

IC

IC
An

IC

IC

IC
An

Figure 4–Functional properties of full-fat (white bars) and defatted amaranth flours (gray bars) from different lines/cultivars.

Vol. 79, Nr. 7, 2014 r Journal of Food Science C1275

 Functional properties of amaranth flours . . .

PCA loading plot as FC and FS showed positive relation with 2.65 g/g) reported in literature (Oshodi and Ekperigia 1989; Kaur
protein content and negative with lipid content (Figure 3). PCA and Singh 2005; Siddiq and others 2010). DAFs had higher WAC
analysis also revealed a strong positive relationship between foam- (2.49 to 2.62 g/g) than AFs (Figure 4). Possibly, the defatting of
ing properties and ash content of the flours. A positive correlation flours resulted into removal of oil that might otherwise hinder the
of FC with protein and ash content, while negative with fat con- absorption of water by hydrophilic side chains. The improvement
tent was reported for pulse protein concentrates (Toews and Wang in WAC following defatting had also been reported for chickpea
2013). protein concentrate and cashew kernel flours (Alobo and others
C: Food Chemistry

EAI and ESI are index of the emulsifying properties of pro- 2009; Toews and Wang 2013). Improved WAC of DAFs makes
teins. The EAI reflects ability of the proteins to rapidly adsorb them suitable for applications in the bakery products, where a
at the oil/water interface during the formation of the emulsion, high WAC of flour would enable baker to add more water to the
while ESI reflects the ability of the protein to maintain a sta- recipe so as to improve dough handling characteristics and main-
ble emulsion. The EAI and ESI of AFs ranged between 33.8 to tain freshness in the bread (Wolf 1970). FAC of flours determines
41.8 m2 /g and 68 to 115 min, respectively. The EAI of AFs was, mouthfeel and flavor retention of products. FAC of AFs ranged
however, lower than soybean proteins (56 m2 /g) reported in lit- between 1.88 and 1.95 g/g that compared favorably with that of
erature (Hojilla-Evangelista and others 2004). The DAFs showed chick pea (1.05 to 1.24 g/g) and sorghum flours (1.72 to 1.85 g/g)
EAI and ESI ranging between 31.3 to 40.9 m2 /g and 59.9 to reported in literature (Elkhalifa and others 2005; Kaur and Singh
102.5 min, respectively. The defatting decreased the EAI and ESI 2005). The FAC of flours is due to the interactions between the
for some of the DAFs (Figure 4). This may partly be ascribed to nonpolar amino acid side chains and hydrocarbon chains of lipids.
higher protein content of DAFs as compared to the counterpart The defatting of the flours did not bring significant (P > 0.05)
AFs. A greater degree of unfolding of polypeptides might have change in FAC (Figure 4). WAC and FAC showed positive relation
occurred in AFs having lower protein content than DAFs, dur- with FC and FS on the PCA loading plot (Figure 3); however,
ing the shearing involved in the emulsifying process and this was the relation was stronger with WAC, suggesting that ability of
aided by hydrophobic association of the peptide chains with the flours to hydrate and bind water could influence their functional
lipid droplets therefore much greater volume of surface area of properties.
protein was made available that may have resulted into enhanced
emulsifying efficiency (Tsai and others 1972). This relation was
SDS-PAGE
also evident as negative relationship between emulsifying proper-
The electrophoretic profiles of the flours from different ama-
ties and protein content on PCA loading plot. Chove and others
ranth cultivars are shown in Figure 5. The AF proteins were char-
(2001) also reported that the emulsifying properties of proteins
acterized by a number of polypeptide subunits between 97 and
decreased as protein concentration was increased.
7 kDa, with main polypeptides of molecular mass (MM) around
WAC is the ability of flour to hold water against gravity wherein
58, 37, 33, 31, 23, and 16 kDa (Figure 5). The electrophoretic
proteins and carbohydrates enhance the WAC of the flours by
profile is similar to those reported in the literature for amaranth
providing hydrophilic parts like polar and charged side chains
proteins. Castel and others (2012) reported major bands of 56.4,
(Pomeranz 1985). The WAC of AFs ranged between 2.09 and
37.2, 35.3, 28.6, 27.2, and 26.2 kDa in DAF from A. mantegazz-
2.43 g/g that were higher than soybean (1.3 g/g), chick pea
ianus seeds. Gorinstein and others (2005) also found several bands
(1.33 to 1.47 g/g), and was comparable with dry bean (2.23 to
in the range of 97 to 14 kDa in the electrophoretic profiles of
A. hybridum, A. cruentus, and A. hypochondriacus. The identifica-
tion of bands in amaranth flour protein profile is difficult because
of the overlap of common bands between the different fractions
IC-540862 DF
Annpurna DF

IC-95341 DF

(Martinez and Anon 1996; Castel and others 2012). However,

IC 35407 DF

PRA-01 DF

IC-540862
PRA-3 DF

Annpurna

IC-95341
IC 35407

the main polypeptide subunit of approximately 59 kDa might

PRA-01
Marker

Marker
PRA-3

be ascribed to Amarantin that is most important component of

amaranth globulins (Singh and Singh 2011). The subunits with
205.0 MW between 37 to 10 kDa might ascribe to amaranth albu-
97.4 mins, glutelins, and alcohol-soluble proteins (Gorinstein and oth-
66.0 ers 1991; Barba de la Rosa and others 1992). AFs from different
lines/cultivars showed similar bands in electrophoretic profiles.
43.0 The defatting did not change the polypeptide composition, as
DAFs showed almost similar banding pattern to AFs (Figure 5).
The results are in accordance with the findings of L’Hocine and
29.0
others (2006), who reported that soy protein isolates prepared from
full-fat meal and defatted meal displayed similar electrophoretic
profiles. The results indicated that protein composition and pro-
20.1
file of AFs was not altered following defatting.

14.3
Conclusion
6.5 The present investigation highlighted the effect of lipids and
proteins on the physicochemical and functional properties of flours
Figure 5–SDS-PAGE profiles of proteins from full-fat and defatted amaranth from different amaranth lines/cultivars. It can be concluded that
flour from different lines/cultivars. Marker: standard protein molecular properties of amaranth flours could be modified by lipid re-
weight marker (×1000 Da), DF, defatted flour. moval/defatting. The defatting improved FV, paste stability during

C1276 Journal of Food Science r Vol. 79, Nr. 7, 2014

Functional properties of amaranth flours . . .
heating and shearing, WAC, and FS of the amaranth flours. The
improvement in these properties was attributed to enhancement in
protein content and reduction in lipid content. Nevertheless, our
results suggested that defatted amaranth flours have potential for
application as functional ingredient in various food applications
due to improved functionality, which vary with line/cultivar.
Acknowledgments
KS wishes to thank CSIR, New Delhi, for providing financial
assistance in the form of a Senior Research Fellowship. The finan-
cial assistance to NS from DST, New Delhi, is also acknowledged.
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