NUTRITIONAL AND FUNCTIONAL PROPERTIES OF AMARANTH GRAIN

FLOUR FRACTIONS OBTAINED BY DIFFERENTIAL SIEVING

Short title: Nutritional properties of amaranth grain flour fractions

Declaration of Interests: None.

KEYWORDS GRAPHICAL ABSTRACT

Differential sieving

Amaranth fractions

Mineral bioaccessibility

Amino acid profile

ABSTRACT

Amaranth grain, a gluten free grain was milled to flour and differentially sieved to coarse and fine fractions. The
whole flour and fractions thereof were analysed for the nutrient composition, antinutrients, total and
bioaccessible minerals, fatty acids and amino acid profile and functional properties of flours. Results indicated
that the fine fraction representing 44% of the whole amaranth flour contained higher protein (19.7%), fat
(8.54%), minerals (3.46%) and dietary fibre content (20.09%) as well as higher overall amino acid profile with
lysine as its major essential amino acid. Linoleic acid (44.8%) in fine flour whereas oleic (29.4%) and palmitic
acid (29.6%) in coarse flour were the predominant fatty acid found in amaranth flour fractions. Minerals were
variedly distributed in analysed fractions as iron was found majorly in fine flour and calcium in coarse flour.
Similar trend in mineral bioaccessibility was observed. The in vitro protein digestibility of amaranth flour
samples ranged from 59.8-72.5%. Functional properties revealed that higher values of water and oil absorption
capacity were characterised in coarse fraction, while whole flour showed higher foaming capacity and stability.
Thus, differentially sieved flour fractions of amaranth grain showed wide distribution of nutrients and in
particular, finer fraction was nutrient dense. It was found to be an excellent source of nutrients and could be
incorporated as functional ingredient in development of nutrient rich products.

1
1. Introduction [6]. Thus, this ancient grain is gaining
Amaranth belongs to family much importance in the present scenario
Amaranthaceae and the word for its rich nutritional content and various
‘Amaranthus’ basically is derived from health promoting properties. Additionally,
Greek word “Anthos” (flower) which amaranth flour presents good film-forming
means everlasting or unwilting. The grain ability and yields films with moderate
amaranth is one among the major solubility, high flexibility, and excellent
pseudocereal grains, which do not belong water vapour barrier properties. These
to grass family but produce fruits and edible films consists of biopolymers which
seeds. This crop has great amount of can be used in food packaging [7].
genetic diversity and has the ability to Milling represents the principle procedure
produce more than one phenotype upon in food industry, where in the outcome-
different environment exposure. With product are further used for the
extreme capacity to adapt to adverse development of finished products. In
growing conditions, the crop can resist milling processes, one of the category is
heat and drought, has no major disease dry milling which separates the outer
reported, and can be easily grown in any fibrous material (seed coat, aleurone and
agricultural land [1]. subaleurone layers) and germ by abrasive
The protein content of amaranth is higher technique to obtain polished grain and by-
than that of most cereals and has been product with high concentration of
reported as an important source of bioactive compounds [8]. Other
bioactive components [2]. The bioactive investigations also report influence of
peptides derived from amaranth protein particle size (74 - 210µm) on the
have shown many biofunctionalities such compositional, functional and rheological
as antimicrobial, antioxidant and properties of lentil flour. And the results
antihypertensive activities [3, 4]. obtained could be beneficial for food
Amaranth protein is also associated with industries to manufacture quality food
potential hypocholesterolemic effect and products with desired functional and
its pure peptides has anti-atherosclerotic rheological characteristics using defined
effect [5]. Amaranth also exerts soluble particle size or fractions [9]. Milled flour
peptides which showed anti-inflammatory is a combination of fine and coarse flour
effects in colonic epithelial cells and with varying nutrient composition and
therefore raised the use of amaranth availability which are bound to the grain
peptides as a potential dietary supplement matrix. Our earlier studies on differential

2
milling of various cereals such as wheat, 2.2. Methods
pearl millet, finger millet and sorghum The amaranth grain was cleaned, dried in
has shown differences in nutritional hot-air oven at 40°C for 4 hr and milled
composition and antioxidant activity [10- into flour in plate mill. The process of
12]. Hence, the present study was differential sieving was carried out by
undertaken with the objective of separating passing the milled whole flour through a
fractions from whole amaranth flour by standard 100 mesh sieve (149 µm) to
differential sieving and analysing these separate into two fractions i.e., fine flour
fractions along with whole flour for and coarse flour. The yield of each flour
nutritional and functional properties. fraction after differential milling was
Further, the in vitro nutrient availability recorded as 44% fine flour and 56% coarse
for selected nutrients was measured in all flour. All the flours were stored in air-tight
flour samples. zip lock pouches at 4°C until further
analysis. A portion of whole flour was
2. Experimental retained as such, which served as control.
2.1. Materials
Amaranth grain (Amaranthus caudatus) 2.2.1. Analysis
was procured in a single lot from local For analysis, differentially sieved flour i.e.
certified organic shop. The analytical fine flour (FF) and coarse flour (CF) along
grade (AR grade) chemicals, solvents, and with whole amaranth flour (WF) were
acids used throughout the experiments, analysed for nutritional composition,
were procured from Sisco Research antinutrient and mineral content, amino
Laboratories Pvt Ltd (SRL), Sd Fine-chem acid and fatty acid profile and functional
Ltd and Merck Life Science Pvt Ltd, properties. The in vitro studies such as
Mumbai, India. In the study, enzymes used protein digestibility and mineral
were pepsin (GRM9155), pancreatin bioaccessibility were also investigated in
(RM3867) and bile salt from Himedia, α- these flours. Same batch of flour were
amylase from Bacillus amyloliquefaciens used in each experiment and all samples
(Lot#SLBF7325V) from Sigma-Aldrich were analysed in triplicates in the
chemicals Pvt Ltd. The dialysis tube following analysis. Triple distilled water
(LA395) with specification 31.71 mm was used in mineral estimation and in
width, 21.5 mm diameter, 2.4 nm pore size vitro studies.
and molecular cut off between 12,000 to Nutrient composition
14,000 were procured from Himedia Co.

3
Differentially sieved flour fraction and WF results are expressed as percent of total
were determined for moisture content by protein solubilized after enzyme
oven drying method, total fat content by hydrolysis.
solvent extraction by Soxhlet method and
protein by Kjeldahl method and the Mineral content and its Bioaccessibility
nitrogen value was multiplied with factor The mineral content of flour samples was
6.25 to obtain protein content [13]. The analysed by ashing the flour in muffle,
soluble, insoluble and total dietary fibre later the ash was digested in concentrated
contents were estimated by rapid HCl over water bath. The process was
enzymatic assay [14]. The carbohydrate repeated twice and further with few drops
content was calculated by difference. of triple distilled water added, the solution
was filtered through ashless filter paper
Antinutritional content and made-up to known volume. The
The phytic acid was estimated by the minerals quantified in ash solution were
method of Thompson and Erdman [15]. iron, zinc, calcium, magnesium,
Phosphorous was converted into phytic manganese, potassium, sodium,
acid using 3.55 as conversion factor. phosphorous, and copper [13]. The
Condensed tannins extracted from flour bioaccessibility of mineral in these flours
fractions using methanol were analysed were also analysed by in vitro method of
colorimetrically by vanillin-HCl method dialysis by simulating gastrointestinal
and results expressed as milligrams of (+)- digestion [18]. The dialysates were
catechin equivalents per 100g [16]. estimated for dialyzable minerals,
In vitro protein digestibility specifically iron, calcium and zinc. The
Akeson and Stahman [17] method was mineral concentration in the prepared ash
followed to determine in vitro digestible solution and in dialysates were measured
protein in amaranth flour fractions. Sample in Inductively coupled plasma optical
containing 100 mg protein was digested emission spectrometer (ICP-OES) (Perkin
with enzyme pepsin and pancreatin in Elmer OptimaTM 8300, MA, USA). This
required pH adjustment. The insoluble spectrometer instrument is an elemental
protein was separated using trichloroacetic analysis technique that identifies, and
acid and the soluble protein was quantifies elements present in trace-level
centrifuged and the supernatant was using emission spectra of a sample.
digested in acid to estimate the protein by Percent bioaccessibility of the minerals
Kjeldahl method. Protein digestibility was computed by comparing the

4
dialyzable mineral with the total mineral condition and expressed as percentage of
present in the flour. relative area.

Functional properties
Amino acid Profile To determine the functional properties, the
Amino acid present in amaranth flour analysed flours were dried at 40°C for
fraction was determined by ion 24hrs to reach equilibrium moisture. Water
chromatography in an Amino acid analyser absorption capacity (WAC) and water
released by hydrolysis with boiling semi- solubility index (WSI) of the differentially
concentrated hydrochloric acid. Biochrom sieved fractions and whole flour was
30+ series of Amion acid analyser from determined by following the procedure
Biochrom Ltd. Cambridge, UK with two- reported by Elhardallou & Walker [21]
channel A/D converter card on a PC using and Anderson, Conway and Griffin, [22]
Software for peak integration was the respectively. WAC is reported as g of
instrument used. The aminograms were water absorbed/100g of dry flour and the
detected at 570 nm and 440nm [19]. weight ratio of dissolved solids in the
supernatant and dry sample reports WSI.
Fatty acid profile Oil absorption capacity (OAC) is the
Liang [20] method was carried out to measure of oil taken up by the known
analyse the fatty acid profile of amaranth amount of flour sample and reported as g
flour fractions. The fatty acid was of oil absorbed/100g of dry flour [23].
measured in gas chromatography (GC) Coffmann and Garcia, [24] method was
fitted with capillary column and flame followed to estimate foaming capacity and
ionization detector into which fatty acid stability in the flour fractions.
methyl esters were fed. GC model used -
GC-2010plus, Shimadzu Corporation, 2.3. Statistical analysis
Tokya, Japan and column used was 30m x Each experiment data were presented as
0.25mm ID, DF-0.25µm thickness. The means ± standard deviation of three
injector and detector temperature were concordant measurements. The significant
maintained at 260 - 280°C. The fatty acids difference comparisons were done by one-
were identified and quantified with way analysis of variance (ANOVA),
reference to the retention time of standard followed by T-test in SPSS 16.0 [25] and
fatty acid methyl esters under similar

5
the statistical significance was defined as P gluten makes it a major gluten-free grain
< 0.05. for incorporation into the diet of celiac
disease patients [29]. The total dietary
3. Results and Discussion fibre was highest in FF (20.1%) followed
3.1. Nutritional composition by WF (15.3%) and CF (12.3%). CF had a
The nutritional composition, higher proportion of IDF, whereas FF and
antinutritional content and invitro WF were rich in SDF. A recent study also
digestible protein of experimental flours reports that amaranth provides higher
are presented in table 1. The analysed flour proportion of soluble fibre than true
with approximately 7 - 9% moisture cereals like wheat and maize which is
content showed nearly 15-20% of protein composed of branched xyloglucans with
and dietary fibre as major nutrient apart majority of di- and trisaccharide side
from carbohydrate. Each flour fractions chains, as well as pectin polysaccharides
differed significantly in terms of protein, [30].
fat, mineral, carbohydrate and total dietary The phytic acid content in flour fractions
fibre content. FF showed higher content of ranged from 1.43 – 2.42 g/100g dw with
protein (19.7%), fat (8.5%) and minerals no significant difference in WF and CF.
(3.5%) and lower carbohydrate content The value of phytic acid in WF is similar
than WF followed by CF on dry weight to the content reported by Akin-Idowu,
basis (dw). Similar values of nutrient Ademoyegun, Olagunju, Aduloju and
content in amaranth WF were reported by Adebo, [31]. Condensed tannins are
Menegassi, Pilosof and Areas, [26] and present in good quantity in the amaranth
Sanz-Penella, Wornkowska, Smietana and grain flour, wherein CF had highest and
Haros, [27]. Another study by Kumar, FF had lowest contents. The IVDP assay is
Dharmaraj, Sakhare and Inamdar, [28] on a widely used method to determine the
fractionation to develop protein and digestibility parameter by mimicking the
mineral rich fraction from amaranth grain, simulated digestive processes in the human
observed higher nutrient content in seed gastrointestinal tract through pepsin-
coat coarser fraction than the finer pancreatin enzyme system to measure the
fraction and the whole flour. Amaranth percentage of proteins hydrolysed by
grain contains high quality protein digestive enzymes [32]. Amongst the
essentially composed of globulin and fraction, the protein digestibility was
albumin with less glutamic acid and highest in FF accounting to 72.5% of total
proline than prolamins, the absence of

6
protein, followed by CF (66.7%) and WF (59.8%).

Table 1: Chemical composition of Amaranth flour fractions per 100g

Flour fractions
Constituent
Whole Fine Coarse
Moisture (g) 7.94±0.07c 8.43±0.07b 8.92±0.06a
Protein (g) 14.7±0.17b (17.06) 16.5±0.18a (19.7) 10.4±0.17c (12.08)
b
Fat (g) 5.83±0.03 (6.33) 7.82±0.06a (8.54) 5.18±0.04c (5.69)
b
Minerals (g) 2.26±0.02 (2.62) 2.90±0.05a 1.87±0.01c (2.18)
CHO (by difference) (g) 50.8 39.8 58.8
Dietary fibre
Total (g) 13.2±0.13b (15.29) 16.8±0.96a (20.09) 10.6±0.36c (12.3)
bc
Insoluble (g) 8.6±0.13 (10.03) 10.8±0.36a (12.85) 8.6±0.23b (10.05)
Soluble (g) 4.5±0.13b (5.26) 6.1±0.6a (7.24) 1.9±0.27c (2.25)
Antinutrients
Phytic acid (g) 1.37±0.06b (1.59) 2.03±0.11a (2.42) 1.23±0.05bc (1.43)
b
Condensed Tannins (mg) 61.4±0.8 (71.2) 42.1±1.7c (50.3) 64.8±1.3a (75.4)
In vitro digestible Protein
Content (g) 10.2±0.22b (11.8) 14.3±0.22a (17.1) 8.1±0.38c (9.4)
Percent (%) 59.8 72.5 66.7
Values reported are mean±standard deviation of triplicate determinations. Mean values with different
alphabetical superscripts are significantly different within a row at p<0.05. Figures in the parenthesis represent
values on dry weight (dw) basis.

3.2. Mineral content and mineral contents of WF & CF. However, in case
bioaccessibility of Ca, CF exhibited higher content (210
The mineral content and its mg/100g dw) with 40% bioaccessibility
bioaccessibility of flour samples are (figure 1). It may be noted that among all
compiled in table 2. The concentration of minerals analysed, CF was richer only in
Fe and its bioaccessibility was higher in calcium content, the difference being
FF i.e. 11.5 mg and 2.99 mg/100g dw significant between flour samples.
respectively, and nearly half of it was Alvarez-Jubete, Arendt and Gallagher,
observed in CF (5.65 mg/100g dw) with [33] in their review on pseudocereals
bioaccessibility of 0.89 mg/100g dw. The nutritive value and their increasing use as
Zn content and its bioaccessibility in flour functional gluten-free ingredients observe
fractions ranged from 2.97 – 4.23 mg and that amaranth, quinoa and buckwheat are
0.66 – 0.88 mg/100g dw respectively, with good sources of important minerals and in
no significant difference between Zn particular they highlight amaranth grain

7
for extremely higher mineral content (Fe, improved hepatic and renal calcium
Ca, Zn and Mg) than other pseudocereals metabolism in Type 2 diabetes mellitus of
and true cereal. The mineral content male Wistar rats. Further analysed
reported in present study is in agreement minerals i.e. Mn, Mg, K, P, and Cu
with values reported in their review and showed significant difference among the
are also comparable with other studies flour fractions presenting higher content in
[27, 34]. The calcium present in amaranth FF. Amaranth flour fractions showed
grain can serve as a crucial source of notable amount of P (381 – 729 mg) > K
supplement and exerts antidiabetic (358 –620 mg) > Mg (219 – 393 mg) per
properties by improving calcium 100g dw. Therefore, partial replacement of
homeostasis in blood, kidney and liver as wheat flour with whole amaranth flour in
described by Kasozi, Namubiru, Safiriyu, bread formulation increased 2-3 fold of
Ninsiima, Nakimbugwe, Namayanja, and both micro- and macro-elements as
Valladares, [35] in their study on reported by Sanz-Penella, Wronkowska,
association of amaranth grain with Soral-Smietana and Haros [27].

Table 2: Mineral content of Amaranth flour fractions

Flour fractions
Minerals
Whole Fine Coarse
Total mineral content (mg/100g)
Fe 6.99±0.14b (8.1) 9.64±0.14a (11.51) 4.85±0.22c (5.65)
bc
Zn 2.56±0.52 (2.97) 3.55±0.13a (4.23) 3.17±0.24b (3.69)
bc
Ca 157±5.6 (182) 159±4.6b (190) 180±6.4a (210)
Mn 2.20±0.02b (2.56) 2.57±0.01a (3.07) 1.91±0.08c (2.23)
b
Mg 249±3.7 (289) 329±7.2a (393) 189±8.0c (219)
b
K 411±2.8 (477) 520±11.2a (620) 307±5.4c (358)
b
P 443±5.1 (513) 610±10a (729) 327±12.8c (381)
Cu 0.51±0.02c (0.59) 0.73±0.06a (0.87) 0.54±0.09b (0.62)
a
Na 5.20±0.76 (6.03) 4.69±0.44ab (5.6) 4.68±0.42abc (5.44)
Bioaccessible minerals (mg/100g)
Fe 0.86±0.01b (1.00) 2.51±0.01a (2.99) 0.76±0.01c (0.89)
c
Zn 0.57±0.0 (0.66) 0.69±0.0a (0.83) 0.67±0.0b (0.78)
b
Ca 51±0.12 (59) 43±0.01c (52) 73±0.01a (85)
Values reported are mean±standard deviation of triplicate determinations. Mean values with different
alphabetical superscripts were significantly different within row at p<0.05 and figures in the parenthesis
represent values in dry weight (dw) basis

8
 Amaranth flour samples
40.4 WF
45
40 32.5 FF
35
26 27.1
30 22.4 CF
25 19.6 21.2
15.7
%

20 12.3
15
10
5
0
Fe Zn Ca

Figure 1: Percent mineral bioaccessibility in amaranth flour samples

and FF were not significantly different.

These values are nearly similar with those
found by Tang, Li, Chen, Zhang, Liu,
3.3. Fatty acid profile
Hernandez, Draves, Marcone, and Tsao,
Table 3 presents the fatty acid profile of
[36], reporting C18:2ω-6 as the main
amaranth flour samples. The predominant
PUFA (37.11-45.92%) and MUFA C18:1
fatty acid found in amaranth grain were
was the second prominent fatty acid
linoleic acid ranging from 37.2 – 44.8%
ranging from 22.72 -31.76% in different
which is a polyunsaturated fatty acid
varieties of amaranth analysed. In our
(PUFA), oleic acid 27.9 – 29.4% being a
study, fatty acids which are present in
monounsaturated fatty acid (MUFA) and
lower amount are linolenic acid (0.58 –
palmitic acid 26.5 – 29.6%, a saturated
1.01% dw), palmitoleic acid (0.5 – 0.6%
fatty acid. Another saturated fatty acid
dw) and myristic acid (0.41-0.77% dw).
present in smaller amount was stearic acid
Our results are also similar to reported
ranging from 5.97 – 7.09% dw. On
values by Shukla, Srivastava, Suneja,
comparing between amaranth flour
Yadav, Hussain, Rana, and Yadav, [37].
fractions, CF constituted higher amount of
Amaranth flour fraction also displayed a
palmitic acid (29.6%), stearic acid
small amount of omega-3 fatty acid i.e.
(7.09%), oleic acid (29.4%), linolenic acid
Eicosapentaenoic acid (EPA) and
(1.01%), arachidic acid (1.91%), and
Docosahexaenoic acid (DHA). The EPA
behenic acid (2.2%) whereas FF showed
content was highest in FF (3.43% dw) and
more of linoleic acid (44.8%) on dw. The
lowest in CF (1.45% dw) and vice versa in
stearic acid, linolenic acid, arachidic acid
case of DHA i.e. CF has highest content
and palmitoleic acid content present in WF
(1.45% dw) and least in FF (1.13% dw).

9
So far, no literature study has reported amino acids which human body cannot
EPA and DHA content in fatty acid profile synthesise in adequate amount under
of amaranth flour. Thus, it can be said that specific physiological or pathological
amaranth provides essential fatty acids in conditions needs to be taken through diet
significant amount and imparts various only. The quantified amino acid profile of
health beneficiary effects. amaranth flour samples is represented in
WHO, [39] reports nutritional importance table 4, which show varying quantity of
of essential amino acids (EAA) as they EAA in different samples. Lysine and
cannot be synthesised in human body, and leucine are the amino acid present in
must be provided through diet. They are higher amount among the EAA ranging
most important for growth and from 0.818 – 1.269% and 0.781-1.25% dw
maintenance of metabolic needs, than non- respectively.
essential amino acids. Apparently, another
category called ‘conditionally essential’

Table 3: Fatty acid profile of Amaranth flour fractions (%)

Flour fractions
Whole Fine Coarse
a
Myristic acid (C14:0) 0.66±0.01 (0.77) 0.35±0.01c (0.41) 0.45±0.03b (0.52)
c
Palmitic Acid (C16:0) 22.9±0.02 (26.5) 23.1±0.08b (27.6) 25.5±0.06a (29.6)
bc
Palmitoleic Acid 0.43±0.01 (0.5) 0.50±0.02a (0.6) 0.51±0.01b (0.59)
(C16:1)
Stearic Acid (C18:0) 5.15±0.03bc (5.97) 5.02±0.05b (6.0) 6.09±0.02a (7.09)
c
Oleic Acid (C18:1) 24.0±0.09 (27.9) 24.2±0.05b (28.9) 25.3±0.03a (29.4)
Linoleic Acid (C18:2) 37.7±0.03b (43.7) 37.5±0.05a (44.8) 31.9±0.05c (37.2)
bc
Linolenic Acid (C18:3) 0.50±0.03 (0.58) 0.53±0.04b (0.63) 0.87±0.04a (1.01)
bc
Arachidic Acid (C20:0) 1.25±0.02 (1.45) 1.23±0.07b (1.47) 1.64±0.01a (1.91)
Behenic Acid (C22:0) 1.62±0.02b (1.87) 1.56±0.05bc (1.86) 1.89±0.01a (2.2)
b
EPA (C20:5) 2.03±0.02 (2.35) 2.87±0.02a (3.43) 1.81±0.01c (2.11)
b
DHA (C22:6) 1.05±0.03 (1.22) 0.94±0.02c (1.13) 1.24±0.01a (1.45)
Others 2.64±0.01b (3.06) 2.09±0.01c (2.5) 2.77±0.01a (3.23)
Values reported are mean±standard deviation of triplicate determinations. Mean values with different
alphabetical superscripts were significantly different within row at p<0.05 and figures in the parenthesis
represent values in dry weight (dw) basis

3.4. Amino acid profile is present in CF. Lastly, methionine,

Next in the series are phenylalanine, valine tryptophan and histidine are present in
and isoleucine present in higher quantity in lower content between the flour fractions.
FF of 0.924, 0.904 and 0.808% Overall, FF has higher content of EAA of
respectively and nearly half of this content 7.34% followed by 5.73% in WF and

10
4.57% in CF on dw basis. Other studies is present in higher amount in amaranth
have also observed the presence of higher flour fractions ranging from 3.671 –
EAA in amaranth flour on comparison 2.097% and it also forms a greater
with corn flour [39] and other proportion of total amino acid content.
conventional cereals [37]. Due to this Overall, the amaranth FF fraction
reason, incorporation of amaranth flour in separated by differential sieving
food products enhances the content of represented rich source of individual
EAA. In particular, it was seen that amino acids, both essential and non-
amaranth incorporation increased 50% of essential and had a much higher content
lysine content in chapatti (Indian than present in WF and CF.
flatbread) fortified with 40% amaranth 3.5. Functional properties
whole flour as compared with the control Values in table 5 represent the functional
whole wheat chapatti [40]. It is also properties of amaranth flour samples.
reported that lysine content in amaranth The CF obtained from differential sieving
meets the daily requirement of infants, showed higher WAC of 174.8±1.2 g and
children and adults. Arginine and glycine FF showed the lowest of 122.2±0.7 g/100g
are among the conditionally EAA present dw conversely the WSI was higher in FF
in higher amount than cystine and proline. (12.2±0.3 g) and least in CF (5±0.3 g).
In non-essential amino acid, glutamic acid

Table 4: Amino acid profile of Amaranth flour fractions (%)

Flour fractions
Whole Fine Coarse
Essential amino acid
Methionine 0.322±0b (0.373) 0.413±0a (0.493) 0.249±0c (0.290)
b
Lysine 0.858±0 (0.995) 1.063±0a (1.269) 0.703±0c (0.818)
b
Threonine 0.517±0 (0.6) 0.648±0a (0.774) 0.414±0c (0.482)
Tryptophan 0.219±0b (0.254) 0.271±0a (0.324) 0.176±0c (0.205)
b
Isoleucine 0.548±0 (0.635) 0.677±0a (0.808) 0.431±0c (0.502)
b
Leucine 0.840±0 (0.974) 1.047±0a (1.25) 0.671±0c (0.781)
Valine 0.611±0b (0.709) 0.757±0a (0.904) 0.484±0c (0.563)
b
Histidine 0.379±0 (0.44) 0.482±0a (0.576) 0.295±0c (0.343)
b
Phenylalanine 0.646±0 (0.749) 0.789±0a (0.942) 0.506±0c (0.589)
Conditionally Essential amino acid
Arginine 1.312±0b (1.521) 1.725±0a (2.06) 0.981±0c (1.142)
b
Cystine 0.317±0 (0.368) 0.402±0a (0.48) 0.248±0c (0.289)
b
Glycine 1.130±0 (1.311) 1.363±0a (1.627) 0.942±0c (1.096)
Proline 0.615±0b (0.713) 0.785±0a (0.938) 0.487±0c (0.567)
Non-essential amino acid
Serine 0.897±0b (1.04) 1.067±0a (1.274) 0.765±0c (0.89)

11
 Alanine 0.540±0b (0.626) 0.677±0a (0.808) 0.425±0c (0.495)
Aspartic acid 1.223±0b (1.418) 1.494±0a (1.783) 1.002±0c (1.166)
Glutamic acid 2.367±0b (2.745) 3.074±0a (3.671) 1.801±0c (2.097)
Methionine+cystine 0.639±0b (0.741) 0.815±0a (0.973) 0.497±0c (0.579)
Total Amino acid 13.98 (16.2) 17.55 (20.95) 11.08 (12.89)
Values reported are mean±standard deviation of triplicate determinations. Mean values with different
alphabetical superscripts were significantly different within row at p<0.05. Figures in parenthesis represent
values on dry weight (dw) basis.
amaranth grains by Sakhare, Inamdar,
Banerji et al., [40] found that WAC
Kumar and Dharmaraj, [42] showed roller
decreases with the increase in the addition
milled amaranth flour produced fractions
of amaranth flour at 20 -50% level from
rich in nutrients with unique functional
1.19 to 1.07 g/g, however, other
properties. The highest foaming capacity
researchers observed the opposite. This
was observed in WF of 8±0.73 ml/100g
difference could be for the fact that
and nearly half of it was reported in both
particle size, extraction rate or
FF and WF which did not show any
composition varies in different flours
significant differences. Similar trend was
based on its nutrient distribution or
observed upon setting aside the foam
composition. The oil absorption capacity
formed by these analysed flours for 30-60
was lowest in FF and did not show any
mins showed slight drop in the stability.
significant differences between WF
Bolontrade, Scilingo and Aňón, [43]
ranging from 85.6 – 87.1 g/100g dw and
reports foams made with amaranth
reports highest in CF (89.1±05 g). The
proteins at acidic pH and low ionic
study by Naik, Dachana, Ramesh and
strength exhibit stability and forms more
Prakash, [41] reports similar trend on
flexible and elastic film. Therefore, these
analysing fractionally milled finger millet
properties provide opportunity to
flour in which overall functional capacities
incorporate amaranth proteins into
of coarser fraction flour were higher than
development of foam-type foods such as
the finer flour fraction. Likewise study on
dessert stuffing and ice creams.
evaluating the roller milling potential of

Table 5: Functional properties of Amaranth flour fractions (%)

Flour fractions
Functional property
Whole Fine Coarse
Water absorption capacity (g/100g) 151.3±0.7b 122.2±0.7c 174.8±1.2a
Water solubility index (g/100g) 8.2±0.4b 12.2±0.3a 5±0.3c
Oil absorption capacity (g/100g) 87.1±1.1b 85.6±1.4bc 89.1±0.5a
Foaming capacity (ml/100ml) 8±0.73a 4.65±0.84b 4.31±0.24bc
30 min 7.24±0.58a 4.63±0.94b 4.01±0.12bc
Foaming stability (ml/100ml)
60 min 7.24±0.58a 4.31±0.24b 3.95±0.24bc

12
Values reported are mean±standard deviation of triplicate determinations. Mean values with different
alphabetical superscripts were significantly different within row at p<0.05.
respectively. Thus, the study indicates
4. Concluding remarks potential utilization of amaranth flour
The present investigation demonstrates fractions as functional ingredients for
that FF contained higher protein, dietary development of nutrient dense dietary
fibre content, and mineral particularly, Fe supplements suitable for different age
and Zn with greater bioaccessibility than groups. The flours could also be used for
WF and CF. The CF showed higher Ca their unique functional properties.
content and bioaccessibility. The fat Acknowledgement
content of amaranth flour samples was 2-3 The authors would like to acknowledge the
fold higher then true cereals. The fatty support provided by Dr. C. K. Girish and
acids of amaranth flours constituted high Evonik Aminolab, Evonik (SEA) Pvt. Ltd.
proportion of essential fatty acids, majorly Singapore to carry out the amino acid
unsaturated i.e. linoleic acid present in profile of amaranth flour samples in the
higher proportion in FF. Contrarily, CF present research work.
had higher saturated fatty acid and MUFA
than other samples. Analysed flours also Conflict of interests
exhibited smaller omega-3 fatty acids (1- The authors declare that there is no
3%). The higher protein quality and conflict of interests regarding the
digestibility in FF showed balanced amino publication of this paper.
acid composition, especially greater in
lysine and glutamic acid content among
essential and non-essential amino acids

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