Journal of Food Composition and Analysis 84 (2019) 103290

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Journal of Food Composition and Analysis

journal homepage: www.elsevier.com/locate/jfca

Original Research Article

12th IFDC 2017 Special Issue – Influence of germination of quinoa T

(Chenopodium quinoa) and amaranth (Amaranthus) grains on nutritional and
techno-functional properties of their flours⋆
⁎
María Dolores Jimenez, Manuel Lobo, Norma Sammán
Faculty of Engineering-CIITED CONICET, National University of Jujuy, Jujuy, Argentina

A R T I C LE I N FO A B S T R A C T

Keywords: The germination process produces changes in grains that may affect the nutritional properties and technological
Food analysis characteristics of their flours. The aim of this work was to determine nutritional changes in quinoa (Chenopodium
Effect of germination on food composition quinoa) and amaranth (Amaranthus) flours, induced by germination under controlled conditions. Proximate
Sprouted grains composition, protein digestibility, starch and fiber were determined by AOAC methods. Total and reducing
Andean grains
sugars (dinitrosalicylic acid method) and protein fractions (SDS–PAGE) were determined in flours. Amylose
Nutrition improvement
content (spectrophotometric method) was determined in starch. Thermal behavior of flour was studied by DSC.
Starch hydrolysis
Protein degradation Protein content and digestibility, and reducing and total sugars were increased by germination. Protein de-
Thermal behavior gradation was observed in fractions with molecular weights higher than 24 kDa in quinoa, while in amaranths
the degradation was in all the molecular weight range 14–66 kDa. Apparent amylose content increased, possibly
due to the formation of dextrins and linear chains of glucans from amylopectin. Gelatinization temperatures
were similar between samples before and after germination. Gelatinization enthalpies of flours were significantly
lower in sprouted than unsprouted grains; also a greater tendency to retrogradation was determined.
Germination improved the nutritional contributions of quinoa and amaranth flours, but the starch content de-
creased and the gel became more unstable, important features if they are to be used as ingredients in food
formulations.

1. Introduction 14% db in quinoa varieties, and from 11 to 21% db in amaranth vari-

Quinoa (Chenopodium quinoa) and amaranth (Amaranthus) grains
grow throughout the Andean region, and have been used by the in-
habitants of South America for centuries. Known as indigenous pseu-
docereals or Andean grains, they can be adapted to different environ-
mental conditions. Pseudocereals are dicotyledonous species which are
not closely related to each other or to the true monocotyledonous
cereals ; the name pseudocereals derives from their production of small
grain-like seeds that resemble in function and composition those of the
true cereals (Valcárcel-Yamani and da Silva Lannes, 2012). There is
very wide genetic variability among the Andean grains, which is re-
flected in the differing nutritional contents and phenotypic character-
istics. The Andean grains are a good source of nutrients with high
protein content (12–16% db) and good lipid profiles (lipid content
between 5% and 10% db). The fraction of dietary fiber varies from 12 to
eties. Moreover, these crops have important amounts of vitamins, mi-
nerals and antioxidant compounds (Repo-Carrasco-Valencia and Serna,
2011; Valcárcel-Yamani and da Silva Lannes, 2012; Carciochi et al.,
2014; Nascimento et al., 2014).
Germination is a biological process that can be applied in an easy
and economical way in order to obtain biotechnologically processed
new food products. The consumption of sprouted products is increasing
because numerous studies have documented their advantages and
health benefits (Moongngarm and Saetung, 2010; Murugkar, 2015).
During the germination process, hydrolytic enzymes are activated
and they are also the newest enzymes synthesized which, along with the
reserve substances in the seed, are mobilized to be used in the initial
growth of the seedling (Bedón Gómez et al., 2013). This process causes
changes in the content and composition of proteins, carbohydrates and
lipids. The proteins are hydrolyzed and consequently their digestibility

⋆
This paper was originally submitted as a poster presentation at the 12th International Food Data Conference held from 11 to 13 October 2017 in Buenos Aires,
Argentina.
⁎
Corresponding author.
E-mail addresses: doloresjimenez4@gmail.com (M.D. Jimenez), mlobo958@gmail.com (M. Lobo), nsamman@fi.unju.edu.ar (N. Sammán).

https://doi.org/10.1016/j.jfca.2019.103290
Received 31 January 2018; Received in revised form 15 September 2018; Accepted 8 August 2019
Available online 13 August 2019
0889-1575/ © 2019 Published by Elsevier Inc.
M.D. Jimenez, et al.
is improved (Chaparro et al., 2010; Omary et al., 2012; Świeca et al.,
2013; Hager et al., 2014; Carciochi et al., 2014; Devi et al., 2015). The
protein profile of grains and legumes has been studied by analysis in
polyacrylamide gel electrophoresis (PAGE) in faba beans (Goyoaga
et al., 2011), lupin (Rumiyati et al., 2012), and quinoa (Valenzuela
et al., 2013), to determine modifications caused by different processes
such as germination, thermal treatments, storage, etc.
Some authors showed that germination of grains and legumes (such
as quinoa, soybean, chickpea, beans, peas, millet, rice and corn) may
decrease the content of antinutrients such as phytates, tannins and in-
hibitory proteases (Omary et al., 2012; Kanensi et al., 2011; Wang et al.,
2015; Jan et al., 2017). Besides, other researchers observed improve-
ments of the polyphenol contents and the antioxidant capacity in
wheat, buckwheat, chickpea, quinoa and amaranth during germination
under certain conditions (Khalil et al., 2007; Paśko et al., 2009).
Simultaneously, germination causes starch degradation by the ac-
tion of amylases producing mainly dextrins, glucose and sucrose that
will be oxidized to produce the energy necessary for the development of
the embryo. Sucrose is the main source of energy storage in the plant
(Bedón Gómez et al., 2013). These modifications can also affect the
functional, rheological, textural and thermal properties of sprouted
grain flours, which are important characteristics if they will be used as
food ingredients.
There are previous records about using cereals, legumes and grains
in food formulations with particular nutritional or technological char-
acteristics (Murugkar, 2015; Yang et al., 2012; Fu et al., 2014; Marengo
et al., 2015; Hallen et al., 2004; Jideani and Onwubali, 2009; among
others), demonstrating that sprouted Andean grains may be used for
this purpose.
The aim of this work was to study changes caused by germination in
nutritional and thermal properties of flours of three varieties of quinoa
(Cica, Kamiri and Inga Pirca) and two varieties of amaranth
(Mantegazzianus and Rosado).
2. Materials and methods
2.1. Andean crops
Quinoa (Cica, Kamiri and Inga Pirca varieties) and amaranth
(Mantegazzianus and Rosado varieties) were obtained from Centro de
Investigación y Desarrollo Tecnológico para la Agricultura Familiar
(CIPAF, Research and Technological Development Center for Family
Agriculture Hornillos, Jujuy, Argentina).
2.1.1. Unsprouted grain flours
The saponin of the quinoa was removed with successive washes
using tap water at room temperature with manual friction, until bitter
taste was eliminated. Amaranth was also washed to eliminate dust.
Both grains were dried in a forced circulation oven at 50 °C until con-
stant weight and then the whole grains were milled in a centrifugal mill
(CHINCAN model FW 100, China). Flours were vacuum-packed in
polyethylene bags and stored at room temperature.
2.1.2. Sprouted grain flours
The washed and saponin-free grains were soaked in tap water (1:5
w/v) for 6 h at room temperature. Water was drained and wet grains
were spread in a thin layer in plastic trays covered with wet filter pa-
pers and incubated under controlled conditions: 22–24 °C and 80–90%
relative humidity in darkness, 24 h for quinoa and 48 h for amaranth
until both germinated grains reached the same length of radical
(1.0–1.5 cm). The germinative capacity was determined according to
Hager et al. (2014), counting germinated grains and expressing it as a
percentage of the total number of grains.
The germinated grains were dried in a forced circulation oven at
50 °C until constant weight. Dried grains were milled in a centrifugal
mill (CHINCAN model FW 100, China). The flours were vacuum-packed
2
Journal of Food Composition and Analysis 84 (2019) 103290
in polyethylene bags and stored at room temperature.
2.2. Proximate composition
The proximate composition of flours was determined by official
techniques AOAC (2017): moisture (method 925.10), ash (method
923.03), fat (method 963.15), total nitrogen (method 920.87); and
N × 6.25 factor was used to calculate total protein content (Chaparro
et al., 2010; Kanensi et al., 2011; Nascimento et al., 2014). Total car-
bohydrates were calculated by the sum of total fiber, total starch and
total sugar. All analyses were carried out in triplicate.
2.2.1. Soluble, insoluble and total dietary fiber
Dietary fiber, both soluble and insoluble, were determined ac-
cording to AOAC Method 991.43 (1995), using the Megazyme assay kit
(Wicklow, Ireland). Samples (1.000 ± 0.005 g) were suspended in
40 mL MES-TRIS buffer (pH 8.2) (MES: 2-(N-morpholino)ethane-
sulfonic acid; TRIS: tris-(hydroxymethyl) aminomethane) and digested
sequentially with 50 μL heat-stable α-amylase (98–100 °C, 30 min),
100 μL protease (60 °C, 30 min), and 200 μL amyloglucosidase (60 °C,
30 min). Digested samples were filtered through fritted glass crucibles;
these insoluble dietary fibers were rinsed with 95% ethanol followed by
pure acetone, and oven dried overnight at 105 °C. Filtrates were mixed
with 95% ethanol (4× volume) to precipitate soluble fibers; after 1 h
they were filtered through tared fritted glass crucibles. Both soluble and
insoluble dietary fiber residues were corrected for protein and ash
content calculated according to the following equations (Eq.1 and
Eq.2):
R1 + R2 + R3
3
− p− A−B
Fiber (Soluble or Insoluble) = 100
m1 + m2 + m3
3 (1)
B= (BR1 + BR2 + BR3)/3 − BP− BA (2)
where mi = sample weight, i = 1, 2 and 3, Ri = residue weight from
mi, i = 1,2, and 3), p = protein weight, A = ash weight, B = blank,
BRi = blank residue, i = 1, 2 and 3; BP = blank protein, and BP =
blank ash.
Total dietary fiber was calculated as the sum of soluble and in-
soluble dietary fiber. Analyses were run in triplicate.
2.2.2. Resistant, digestible and total starch
The resistant and digestible starches were determined using the
Megazyme assay kit (Wicklow, Ireland) following the AOAC method
(2003). Flour samples (100 ± 0.5 mg) were incubated with 4 mL
pancreatic α-amylase (10 mg/mL) solution containing amyloglucosi-
dase (AMG) for 16 h at 37 °C with constant shaking (200 strokes/min);
in order to terminate the reaction, ethanol (99% v/v) was added. The
pellet was separated by centrifugation (1500 g, 10 min) and re-sus-
pended in ethanol (50% v/v); then it was digested with 2 mL KOH (2 M)
in an ice bath with vigorous magnetic stirring. Digested pellet and su-
pernatant were separately incubated with AMG. The D-glucose released
was measured using a glucose oxidase-peroxidase (GOPOD) reagent kit,
and the absorbance was read at 510 nm against the reagent blank (UV-
6300PC, MAPADA, Shanghai). The total starch was calculated as the
sum of resistant and digestible starch. Analyses were run in triplicate.
2.2.3. Reducing and total sugar
2.2.3.1. Reducing sugar. The reducing sugar was determined by the
spectrophotometric method (Miller, 1959) with minor modifications
mentioned by Başkan et al. (2016), based on cupric reduction. DNS
(3,5-dinitrosalicylic acid) reagent was prepared with 1:1:1:0.5 (v/v/v/v)
ratio of: DNS solution (1% w/v), phenol solution (0.2% w/v), NaOH
solution (1.5% w/v) and sodium biftalate (0.15% w/v). Flour samples
(1.000 ± 0.005 g) were suspended with 10 mL distilled water and the
mixture was shaken. Solution was centrifuged (10,000 rpm, 10 min,
M.D. Jimenez, et al.
20 °C). An aliquot (25 μL) was solubilized in 75 μL water and 1 mL DNS
solution was added. The mixture was shaken and incubated in a boiling
water bath for 5 min, then 1 mL sodium potassium tartrate (Rochelle
salt) (40% w/v) was added and it was immediately cooled in ice bath
during 15 min. The absorbance of sample was recorded at 540 nm
against a reagent blank (UV-6300PC, MAPADA, Shanghai). A linear
regression equation was obtained using a D(+)-glucose standard
solution (Sigma-Aldrich, Steinheim, Germany). Analyses were run in
triplicate.
2.2.3.2. Total sugars. Total sugar content was determined by the
method described by Başkan et al. (2016) with slight modifications.
Hydrolysis of sucrose was performed in the supernatant (3 mL),
obtained in point 2.2.3.1, with 0.5 mL of concentrated HCl, 15 min in
a boiling water bath; the mixture was neutralized with 2 M NaOH. An
aliquot (25 μL) was solubilized in 75 μL water and then analysed as
indicated for reducing sugars. A linear regression equation was
obtained using a hydrolyzed sucrose standard solution (Sigma,
Steinheim-Germany). Analyses were run in triplicate.
2.3. Protein fractions
The protein fractions of samples, before and after germination, were
separated by SDS–PAGE. Alkaline proteins extraction of the flours was
performed with NaOH 0.2% w/v (ratio 1:10) and vortexed for 30 min
(Rumiyati et al., 2012). The mixture was centrifuged (8000 g, 25 °C,
5 min). The supernatant was dissolved with sample buffer pH 6.8 (ratio
1:1) containing 10% (v/v) glycerol, 1% (w/v) SDS and 0.05% (w/v)
bromophenol blue. To determine the disulfide bonds, the samples were
run in conditions similar to those above but with the addition of 5% (v/
v) β-mercaptoethanol and heating in a boiling water bath for 5 min
(denaturing and reducing conditions).
Molecular weight (MW) markers from Sigma (6.5–205 kDa) were
used.
The stacking gel had 5%T acrylamide:bisacrylamide (0.049 M
Tris−HCl, pH 6.8) and the separating gel was 10%T and 12%T for runs
without and with β-mercaptoethanol respectively (0.375 M Tris−HCl,
pH 8.8); both contained 1% (w/v) SDS. Running buffer (pH 8.3) was
prepared with 0.025 M Tris−HCl, 0.186 M glycine and 1% (w/v) SDS.
The electrophoresis runs were conducted at a constant voltage of 100 V.
Gels were washed with washing solution of acetic acid (10%) for
10 min and stained with 0.1% (w/v) Coomassie Brilliant Blue R 250
(Sigma, St. Louis, MO) in 10% (v/v) acetic acid for 15 min.
Gels were scanned, and the molecular mass of the bands assessed
using the software GelAnalyzer2010 (J&B Lab. S.A.C, Lima, Peru).
2.4. Protein digestibility
In vitro protein digestibility was determined by AOAC 971.09
method modified by Miller (2002). Defatted flours (1 g) were digested
with 150 mL of pepsin solution in 0.075 M HCl (0.0002% v/v) and in-
cubated in a shaking bath (45 °C, 16 h) (Viking Shaker, Argentina). The
digested samples were vacuum-filtered and washed three times with
water and acetone. In the residue, the nitrogen content was determined
by Kjeldahl method (AOAC 920.87). The value was corrected by a ni-
trogen determination in an HCl solution free of pepsin; then the protein
was calculated by using 6.25 factor. Analyses were run in triplicate.
2.5. Starch
2.5.1. Isolation
Starch was isolated from the flours of the quinoa and amaranth
varieties by alkaline steeping method (Jan et al., 2017) with slight
modifications in the procedure. Defatted flour was dispersed in water
(1:10 w/v) and the pH was adjusted to 9.0 with 2 M NaOH and placed at
room temperature for 24 h. The samples were mixed for 30 min and the
3
Journal of Food Composition and Analysis 84 (2019) 103290
blended slurry was filtered through 400 (37 μm) mesh. The filtrate was
centrifuged (9000 g, 5 °C, 20 min), and the residue was re-suspended in
water and neutralized with 1 M HCl. The resultant suspension was then
filtered through a filter paper. The retained material (starch) was wa-
shed five times for purification with sufficient water and 200 mL of
ethanol (96% v/v) and then dried in air oven at 40 °C 12 h. The dried
starch was milled in a centrifugal mill (CHINCAN FW 100, China). The
starch was vacuum-packed in polyethylene bags and stored at room
temperature.
2.5.2. Amylose
Amylose content was measured by the method described by Juliano
et al. (1981) and mentioned by Lu et al. (2013) with minor modifica-
tions, based on the measurement of the iodine–amylose complex. Starch
samples (100 mg) plus 1 mL of 96% (v/v) ethanol and 9 mL of 1 M
NaOH were kept at room temperature for 24 h, and then distilled water
was added to make 100 mL solution. An aliquot of 0.5 mL was trans-
ferred to a 10-mL volumetric flask containing 5 mL of distilled water,
and 0.1 mL 0.4 M Na2HPO4 buffer (pH 7.2) was added. Then 2 mL of
0.2 g/L iodine solution (I2: 2 g/KI: 20 g/L) and distilled water were
added to make up exactly 10 mL. Spectrophotometer measurements
were made at 620 nm (UV-6300PC, MAPADA, Shanghai) after the
above starch–iodine solution was incubated for 20 min at room tem-
perature. A standard curve was generated using potato amylose–amy-
lopectin (A0512, Sigma Chem. Co., St. Louis, MO). Analyses were run in
triplicate.
2.5.3. Thermal behavior
Thermal properties of flours were determined by differential scan-
ning calorimeter (DSC) using a TA Instrument model Q2000 (TA
Instruments, New Castle, DE). Flour (2.0 mg) was weighed into an
aluminum pan and 10 μL distilled water were added. The pan was
hermetically sealed and equilibrated at room temperature for 1 h, then
scanned at a heating rate of 10 °C/min from 25 to 120 °C with an empty
sealed pan as a reference. Onset (To), peak (Tp), final (Tf) tempera-
tures, and enthalpy of gelatinization (ΔH) were determined by
Universal Analysis 2000 Program (TA Instruments). After cooling, the
scanned sample pans were placed in a refrigerator at 4 °C for 21 days.
Retrogradation properties were measured by rescanning the sample
pans under the same conditions. The percentage of retrogradation (R%)
was calculated as (ΔHr/ΔH) × 100. The experiments were carried out
in triplicate.
2.6. Statistical analysis
Results were expressed as mean ± SD. One-way analysis of var-
iance (ANOVA) was used to compare the means. Differences were
considered significant at p < 0.05. Statistical analyses were performed
with Statistix for Windows version 9.0 (Analytical Software,
Tallahassee, FL).
3. Results
Quinoa and amaranth grains possessed high germinative capacity.
Quinoa reached a length of radicle of 1.0–1.5 cm in 24 h while the
amaranth reached the same length after 48 h.
3.1. Proximate composition
Table 1 shows the proximate composition of unsprouted and
sprouted grains. The germination increased the protein content sig-
nificantly in all varieties of quinoa and amaranth (p < 0.05), except in
the Inga Pirca variety; in which the protein content did not undergo
significant change. On the other hand, the lipid content was not mod-
ified significantly with the germination. The ash content presented a
slight increase with the germination, only significant (p < 0.05) in the
M.D. Jimenez, et al. Journal of Food Composition and Analysis 84 (2019) 103290

Table 1 Table 2
Proximate composition of grain flours. Protein digestibility.
Grain variety Ash Protein Fat Grain variety Protein digestibility (%)

e f ab
Quinoa Cica 2.16 ± 0.10 12.74 ± 0.31 7.48 ± 0.14 Quinoa Cica 61.27 ± 0.38b
Sprouted Quinoa Cica 2.26 ± 0.01de 14.31 ± 0.14bcd 5.61 ± 1.94b Sprouted Quinoa Cica 87.11 ± 0.41a
Quinoa Kamiri 2.36 ± 0.02d 12.97 ± 0.22ef 6.53 ± 0.18ab Quinoa Kamiri 63.24 ± 3.58b
Sprouted Quinoa Kamiri 2.58 ± 0.14c 15.78 ± 0.12a 5.99 ± 0.08b Sprouted Quinoa Kamiri 72.89 ± 5.30b
Quinoa Inga Pirca 2.25 ± 0.02de 14.55 ± 0.24bc 6.75 ± 0.27ab Quinoa Inga Pirca 63.23 ± 1.21b
Sprouted Quinoa Inga Pirca 2.37 ± 0.02d 14.00 ± 0.31cd 6.79 ± 0.05ab Sprouted Quinoa Inga Pirca 72.89 ± 0.24b
Amaranth Rosado 2.88 ± 0.05b 12.51 ± 0.12f 7.78 ± 0.02ab Amaranth Rosado 72.33 ± 3.69b
Sprouted Amaranth Rosado 3.03 ± 0.05b 13.94 ± 0.21cd 8.81 ± 0.33a Sprouted Amaranth Rosado 86.73 ± 2.19a
Amaranth Mantegazzianus 3.05 ± 0.05b 13.74 ± 0.16de 6.66 ± 0.14ab Amaranth Mantegazzianus 71.57 ± 4.35b
Sprouted Amaranth 3.53 ± 0.06a 14.69 ± 0.19bc 7.00 ± 0.25ab Sprouted Amaranth Mantegazzianus 85.34 ± 2.56a
Mantegazzianus
Values are means ± standard deviations of data from triplicate analysis.
Values are means ± standard deviations (g/100 g db) from triplicate analysis. Values followed by different superscript letters in the same column are sig-
Values followed by different superscript letters in the same column are sig- nificantly different (p < 0.05).
nificantly different (p < 0.05).
other authors (Bedón Gómez et al., 2013; Janssen et al., 2017). They
Kamiri varieties for quinoa and Mantegazzianus for amaranth. also differentiated the protein fractions of quinoa into albumins
(14–66 kDa), globulins (14–36 kDa), prolamins (24, 29, 36 and 55 kDa)
3.2. Protein fractions and glutelins (14–55 kDa); and albumins1 (20–36, 45, 66 kDa), albu-
mins2 (29, 36, 45, 66 and > 66 kDa) and globulins (29 a > 66 kDa) for
The polyacrylamide gel electrophoresis (SDS–PAGE) analysis of the amaranth.
protein extracts of unsprouted and sprouted quinoa and amaranth is Some protein bands are smaller in the profiles of germinated grain
shown in Fig. 1a and b, respectively. Proteins of molecular weights samples, indicating enzymatic hydrolysis of proteins during
between 14 and 66 kDa were identified (Fig. 1a and b); this agrees with

Fig. 1. Representative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). (a and b) without and (c and d) with β-mercaptoethanol. P: MW
standard; 1: Quinoa Cica; 2: Quinoa Kamiri; 3: Quinoa Inga Pirca; 4: Sprouted Quinoa Cica; 5: Sprouted Quinoa Kamiri; 6: Sprouted Quinoa Inga Pirca; 7: Sprouted
Amaranth Mantegazzianus; 8: Sprouted Amaranth Rosado; 9: Sprouted Amaranth Mantegazzianus; 10: Sprouted Amaranth Rosado.

4
M.D. Jimenez, et al. Journal of Food Composition and Analysis 84 (2019) 103290

germination.
carbohydrate
The electrophoretic profile with β-mercaptoethanol showed new
bands (Fig. 1c and d) with respect to electrophoretic profile without this
73.80
74.24
74.74
71.75
72.81
72.88
72.77
71.44
73.79
71.22
Total

reducing agent (Fig. 1a and b), which means that the mercaptoethanol
broke the disulfide bonds, evidencing their presence in the proteins of
quinoa and amaranth grains, before and after germination.
0.56a
0.36a
0.45a
0.85a
0.79a
1.01a
0.52a
0.47a
0.50a
1.01a
3.3. Protein digestibility in vitro
±
±
±
±
±
±
±
±
±
±
Soluble
fiber

3.85
3.55
3.71
3.75
3.28
2.55
3.54
4.17
2.99
3.44
Table 2 presents the protein digestibility in vitro of sprouted and
unsprouted grain flours. The protein digestibility was improved sig-
nificantly with germination in all varieties of both grains. This increase
11.29 ± 1.44abc
11.85 ± 1.32ab
12.00 ± 1.93ab

11.85 ± 0.14ab
9.37 ± 0.28bcd

12.19 ± 0.27a
8.86 ± 0.94cd
9.02 ± 0.29cd

was also observed by Chaparro et al. (2010) for quinoa and amaranth
7.99 ± 0.46d

7.91 ± 0.25d

grains. Omary et al. (2012) observed increments in protein digestibility

Insoluble

with the germination of millet, corn and sorghum, among others.

fiber

3.4. Carbohydrates
0.32abc

0.89abc
0.54abc
1.17abc

Table 3 shows the carbohydrates composition of the different flours.

0.39ab
0.51bc

1.23a
0.96a

0.57a
0.35c

The resistant starch did not show significant variations by germination,

±
±
±
±
±
±
±
±
±
±

while the digestible starch decreased significantly in all the varieties

11.83
12.92
11.62
12.61
12.29
14.40
15.54
15.46
15.18
15.28
Total
Fiber

studied. This was observed by other authors (Omary et al., 2012; Wu

et al., 2013; Hager et al., 2014) in legumes, millet, rice, amaranth and
quinoa.
The total, soluble and insoluble fiber contents did not show sig-
Reducing sugar

18.97 ± 1.03d
34.02 ± 0.87b

42.75 ± 1.52a
29.73 ± 1.18c

29.41 ± 1.41c
5.41 ± 0.47e

4.42 ± 0.05e

4.75 ± 0.17e

5.48 ± 0.45e

5.02 ± 0.05e

nificant variations during germination, except in the variety of quinoa

Inga Pirca where insoluble fiber increased (p < 0.05). Insoluble
dietary fiber was the main fraction of total dietary fiber in all samples,
since it represented from 67 to 80%.

3.5. Amylose
0.09ef

0.14ef

0.33ef
0.68d
0.60b
0.77a
0.25e

0.75c

0.66c
0.30f

Table 4 shows the amylose content in the starches of all studied

±
±
±
±
±
±
±
±
±
±

Values followed by different superscript letters in the same column are significantly different (p < 0.05).
17.58
52.28
14.05
35.57
14.73
42.98
14.62
29.63
13.23
39.23
sugar

grain samples. It can be observed that the apparent amylose increased

Total

significantly (p < 0.05) in all grains with germination. The increase

was between 13 and 65% in quinoa while in amaranth it was higher
than 100% for both studied varieties.
43.69 ± 0.28ab

45.10 ± 1.55ab

41.27 ± 1.76bc
12.86 ± 1.95ef
22.97 ± 0.55d

21.67 ± 1.28d
48.51 ± 1.64a

15.00 ± 1.61e
37.57 ± 0.57c
8.56 ± 0.55f
Digestible

3.6. Thermal behaviors

Starch

Table 5 shows thermal behavior of quinoa and amaranth flours

before and after germination.
Values are means ± standard deviations (g/100 g db) from triplicate analysis.

The gelatinization temperatures (initial, peak and final) did not

0.35ab
0.09bc
0.02d
0.01d
0.01d
0.13d
0.03d
0.02d
0.02a

0.35c

show significant variations between unsprouted and sprouted grain

flours, which indicated that the cooking temperature of the grains be-
Resistant

±
±
±
±
±
±
±
±
±
±
Starch

fore and after germination would be similar. However, the gelatiniza-

0.70
0.47
0.56
0.59
0.69
0.50
5.05
4.68
4.12
3.86

tion energy (ΔH) was significantly less in the sprouted grain flours. The
reduction of the enthalpy (ΔH) with germination (Table 5) could be
44.39 ± 0.30ab

45.79 ± 1.58ab

45.38 ± 1.67ab
15.50 ± 1.63d

16.71 ± 1.59d
42.61 ± 0.55b
49.07 ± 1.63a
23.57 ± 0.68c

26.35 ± 0.93c

Table 4
9.04 ± 0.54e

Apparent amylose content of starch.

starch

Grain variety Amylose

Total
Carbohydrates composition of grain flours.

Quinoa Cica 9.45 ± 0.04d

Sprouted Quinoa Cica 14.24 ± 0.11a
Quinoa Kamiri 7.45 ± 0.26e
Sprouted Amaranth Mantegazzianus

Sprouted Quinoa Kamiri 12.32 ± 0.15b

Quinoa Inga Pirca 7.76 ± 0.04e
Sprouted Quinoa Inga Pirca 8.74 ± 0.15d
Sprouted Quinoa Inga Pirca

Sprouted Amaranth Rosado

Amaranth Mantegazzianus

Amaranth Rosado 1.45 ± 0.26f

Sprouted Quinoa Kamiri

Sprouted Amaranth Rosado 8.76 ± 0.41d

Sprouted Quinoa Cica

Amaranth Mantegazzianus 1.74 ± 0.15f

Quinoa Inga Pirca

Amaranth Rosado

Sprouted Amaranth Mantegazzianus 11.47 ± 0.22c

Quinoa Kamiri
Grain variety

Quinoa Cica

Values are means ± standard deviations (g/100 g db) from triplicate ana-
Table 3

lysis.
Values followed by different superscript letters in the same column are
significantly different (p < 0.05).

5
M.D. Jimenez, et al. Journal of Food Composition and Analysis 84 (2019) 103290

Q.: Quinoa; A.: Amaranth; Sp.: Sprouted; To: initial gelatinization temperature; Tp: peak gelatinization temperature; Tf: final gelatinization temperature; ΔH: gelatinization enthalpy; To(r): initial retrogradation
related to the reduction in the starch content (Table 3). The retro-

14.04 ± 0.03d

13.91 ± 0.23d

14.19 ± 0.81d

15.43 ± 0.33d
21.02 ± 1.35b

25.67 ± 0.49a
18.28 ± 0.09c

9.97 ± 0.45e

6.09 ± 0.06f

6.78 ± 0.36f
gradation enthalpies (ΔHr) in germinated grain flours were less than in
non-germinated only in the Cica quinoa variety and in the Rosado
amaranth variety, while in other varieties ΔHr did not suffer significant
R%

variations with germination.

0.02bcd
4. Discussion
0.02cde
0.02de
0.02cd
0.02bc

0.01bc
0.02bc
0.02ef
0.02b
0.04a

The ash content increased slightly with the germination of Kamiri

ΔHr (J/g)

±
±
±
±
±
±
±
±
±
±
quinoa and Mategazzianus amaranth. The mineral content of tap water
1.21
0.89
0.76
0.69
0.57
0.52
0.63
0.40
0.27
0.75
used for germination (wash and soaked) may have contributed to the
ash content (Suma and Urooj, 2014) or it could be explained by a loss of
dry matter which produced an apparent increase of ash content (Khalil
Values are means ± standard deviations from triplicate analysis. Values followed by different superscript letters in the same column are significantly different (p < 0.05). et al., 2007). In other varieties, the ash content did not present sig-
a

0.97a
1.67a
1.53a
1.04a
1.02a
1.22a
1.59a
2.06a
0.91a
2.14

nificant variation. These results were in agreement with other authors

Tf(r) (ºC)

±
±
±
±
±
±
±
±
±
±

(Omary et al., 2012).

64.88
64.84
66.84
63.75
60.89
60.18
64.26
63.55
64.35
60.71

The lipid content did not show variations during the germination
and the obtained values agreed with the ones observed by Omary et al.
temperature; Tp(r): peak retrogradation temperature; Tf(r): final retrogradation temperature; ΔHr: retrogradation enthalpy; R%: percentage of retrogradation.

(2012). However, other research showed a reduction of the lipid con-

tent with germination, as lipids would have been used as an energy
a

1.67a
0.80a
1.37a
1.61a
0.69a
1.23a
0.57a
1.63a
1.82a

source (Rumiyati et al., 2012; Devi et al., 2015).

1.34

Protein content increased during germination, from 12 to 22% db in

Tp(r) (ºC)

±
±
±
±
±
±
±
±
±
±

quinoa and between 7 and 11% db in amaranth. Apparent increases in

51.57
52.13
50.14
50.70
50.44
49.45
49.27
47.52
53.31
50.69

protein content (and other nutrients) could be explained by a loss of dry

matter, particularly through carbohydrates oxidation and the release of
CO2, H2O and small amounts of ethanol. These results agree with those
shown by Khalil et al. (2007) and Omary et al. (2012). On the other
a

1.39a
1.01a
1.11a
0.80a
0.95a
1.21a
1.10a
1.24a
0.60a
Retrogradation

0.32

hand, Kanensi et al. (2011) did not report significant changes in protein
To(r) (ºC)

±
±
±
±
±
±
±
±
±
±

content in the germination of amaranth grains, with 5 h of soaking and

39.58
40.79
38.96
39.53
37.96
37.86
38.98
39.15
41.45
40.15

24 h of germination; but by varying the germination time to 48 h, these

authors found that the protein content increased significantly.
In addition, electrophoretic profiles showed that proteins, especially
those of high molecular weight, were hydrolyzed and mobilized during
3.66 ± 0.05def
10.35 ± 0.25a

3.98 ± 0.03de
4.87 ± 0.13cd

germination. High molecular weight proteins were enzymatically de-

3.28 ± 0.03ef

2.92 ± 0.07ef
b

5.46 ± 0.11c

5.71 ± 0.11c

2.59 ± 0.08f
8.62 ± 0.28

graded with germination; in quinoa the proteins greater than 24 KDa

ΔH (J/g)

were mainly degraded, while in amaranth the degradations were pro-

duced in all fractions between 14 and 66 KDa. Degradation is observed
by the reduction of band width, which would involve less protein of
those molecular weights in the samples of sprouted grains. The decrease
in the molecular size of proteins could be related to the increased in
a

2.53a
2.29a
1.95a
1.31a
0.98a
2.42a
1.95a
0.80a
2.22a
0.90

protein digestibility, which was 15–42% in quinoa and 19–20% in

amaranth (Chaparro et al., 2010; Omary et al., 2012).
±
±
±
±
±
±
±
±
±
±
Tf (ºC)

81.53
81.58
81.74
81.32
81.69
81.37
86.26
81.10
88.38
82.33

Quinoa and amaranth proteins have disulfide bonds (Valcárcel-

Yamani and da Silva Lannes, 2012); germination did not break these
unions completely as shown in Fig. 1, comparing the protein profile in
the SDS–PAGE without and with β-mercaptoethanol. If the flours are
a

1.70a
1.89a
2.37a
0.88a
0.69a
2.12a
1.97a
0.87a
1.41a

used in a food formulation where the viscosifying and gelling properties

0.46

will stabilize the food matrix (Liu et al., 2016), the presence of disulfide
±
±
±
±
±
±
±
±
±
±
Tp (ºC)

bonds is important. In addition, the existence of disulfide bonds in-

65.24
67.93
67.57
67.66
67.52
68.82
68.32
70.37
72.19
70.68

dicates the presence of essential sulfur amino acids (methionine and

cysteine) which would nutritionally improve the formulated products
(Valcárcel-Yamani and da Silva Lannes, 2012).
Total, soluble and insoluble fiber contents did not change by ger-
a

1.83a
1.62a
1.50a
1.31a
1.18a
1.41a
1.30a
2.02a
1.67a
1.22
Gelatinization

mination (except in quinoa Inga Pirca variety), a result which coincided

with those of some authors (Repo-Carrasco-Valencia and Serna, 2011;
±
±
±
±
±
±
±
±
±
±
To (ºC)

60.83
61.09
62.20
62.62
62.18
62.04
64.05
65.03
66.14
66.80

Quinto et al. 2015). Nevertheless, other authors have reported that the
fiber content increased with germination (Moongngarm and Saetung,
2010) while other researchers have reported that the fiber decreased in
Thermal behavior of flours.

germination of buckwheat, rice, sorghum and amaranth (Omary et al.,

A. Mantegazzianus Sp.

2012). Dueñas et al. (2016) reported a notable increase during germi-

nation in the level of insoluble and total dietary fiber in beans and, on
A. Mantegazzianus
Q. Inga Pirca Sp.

the contrary, germinated lentils showed a decrease in the total dietary

A. Rosado Sp.
Q. Kamiri Sp.
Grain variety

Q. Inga Pirca

fiber as a consequence of the marked decrease in the insoluble dietary

Q. Cica Sp.

A. Rosado
Q. Kamiri

fiber. These results are in concordance with Gong et al. (2018) who
Q. Cica
Table 5

indicated an increase in the soluble fiber and a decrease in the insoluble

fiber upon germination of corn. Dueñas et al. (2016) indicated that an

6
M.D. Jimenez, et al.
increase in soluble fiber may be a consequence of the rise of cellulosic
glucose due to the metabolic reactions undergone by seeds during
germination. Thus the effect of germination in total, soluble and in-
soluble dietary fiber content depends on the type of grain. These grains
exhibited different behaviors during the process, due to the different
structures and compositions of the cell wall as well as the conditions of
germination process (Omary et al., 2012; Dueñas et al., 2016; Gong
et al., 2018).
Digestible starch decreased during germination by action of amy-
lolytic enzymes. The partial hydrolysis of starch produced a significant
increase (p < 0.05) in reducing and total sugars higher than 100%;
these results are similar to those reported by Hager et al. (2014) for
quinoa and amaranth. The starch hydrolysis would increase its digest-
ibility, which could produce an increase in the glycemic and in-
sulinemic response (Bedón Gómez et al., 2013; Świeca et al., 2013). On
the other hand, the resistant starch did not suffer significant variations
because it cannot be attacked by enzymes during germination.
The amylolytic enzymes, α-amylase and β-amylase, hydrolyzed the
starch, producing mainly dextrin, glucans, sucrose, fructose, glucose,
maltose, oligomaltose and maltotriose. The increment of apparent
amylose content (between 13–65% in quinoa grains and over 100% in
amaranth grains) could originate from the hydrolysis of primary amy-
lopectin that releases linear branches of glucan chains and dextrins
which are reactive to the iodine just like the amylose (Juliano et al.,
1981). This assumption is consistent with the results obtained by Wu
et al. (2013), who determined the amylose/amylopectin ratio in flours
of different cultivars of brown rice after different germination times, by
enzymatic method amylose–amylopectin assay kit. These authors re-
ported that amylose and amylopectin gradually decreased with germi-
nation.
Gelatinization enthalpy decreased probably by starch hydrolysis
with increased amylose/amylopectin ratio due to greater degradation
of the amylopectin during germination. These results are similar to the
ones obtained by Wu et al. (2013), who studied the changes in rice
starch after germination; they noted a slight decrease in gelatinization
enthalpy, probably due to an increase in the amylose/amylopectin ratio
during germination.
Enthalpy of retrogradation did not change in the same way for all
varieties of grain flours before and after germination. This measure
depends on several variables, such as the molecular ratio of amylose/
amylopectin, structures of the amylose and amylopectin molecules
(which are determined by the botanical source of the starch), starch
content, the presence and concentration of other components
(Fennema, 2017). On the other hand, germination increased the ret-
rogradation (R%) in quinoa and amaranth caused by the higher amy-
lose/amylopectin ratio (Table 4). Although R% increased, it is im-
portant to emphasize that the total content of starch in sprouted grains
was lower (Table 3). Therefore, the amount or crystalline structures
capable of retaining water was also lower. Yu et al. (2009) reached the
same conclusions in the study of the impact of amylose content on
retrogradation of rice starch.
5. Conclusions
Germination improved the nutritional contributions of the quinoa
and amaranth flours, as the proteins and carbohydrates were hydro-
lyzed during germination, and the protein hydrolysis improved their
digestibility.
The decrease in the starch content and the changes in its quality,
produced by germination, generated alterations in the gelatinization
enthalpy, but the cooking temperatures were not modified. However,
germination increased the retrogradation of the flours, due to the
structural modifications that carbohydrates and proteins undergo. In
this way, the changes in the starch and proteins during germination
could surely produce changes in the rheological and textural behavior
of the products elaborated with these flours. and probably in the
7
Journal of Food Composition and Analysis 84 (2019) 103290
sensory properties of formulated food with germinated grain flours.
Thus, as this study shows, germination is a low-cost technology that
improves the nutritional contributions of quinoa and amaranth grains.
However, if these flours are used as ingredients in food formulations,
modifications of the thermal behavior and rheological properties must
be considered.
Declaration of Competing Interest
The authors declare do not have conflict of interest of any kind.
Acknowledgements
The authors would like to thank Silvia Rebeca Chañi for her tech-
nical assistance in the determinations in differential scanning calori-
meter (DSC), in National University of Jujuy (Jujuy, Argentina).
This work was supported by the Consejo Nacional de
Investigaciones Científicas y Técnicas (CONICET), Ministerio de Ciencia
y Tecnología, Argentina.
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