foods

Article
Effect of Germination on the Nutritional Properties, Phytic Acid
Content, and Phytase Activity of Quinoa (Chenopodium quinoa
Willd)
Pedro Maldonado-Alvarado 1 , Darío Javier Pavón-Vargas 1 , Juan Abarca-Robles 1,2 , Silvia Valencia-Chamorro 1
and Claudia Monika Haros 2, *

1 Department of Food Science and Biotechnology, Escuela Politécnica Nacional, Quito 17-01-2759, Ecuador
2 Cereal Group, Instituto de Agroquímica y Tecnología de Alimentos IATA-CSIC, 46980 Valencia, Spain
* Correspondence: cmharos@iata.csic.es; Tel.: +34-96-390-0022

Abstract: The aim of this study is to evaluate the effect of desaponification, soaking, germination,
and refrigerated storage on the phytase activity, phytic acid content, and nutritional properties of
three varieties of quinoa: white, red, and black. Desaponification and soaking reduced the number of
minerals and the nutritional content. Germination of the seeds was carried out in the desaponified
samples. The nutritional values, phytase activity, and phytic acid content of quinoa were measured
after 6 h of soaking and then at 4 and 7 days during germination plus 7 days of refrigerated storage
(4 ◦ C). Germination increased the fibre and protein content as well as the iron, zinc, and calcium
content. Germination significantly increased the phytase activity in all varieties and decreased
the phytic acid content. The phytic acid content decreased during germination from 32 to 74%.
Refrigerated storage had no significant effect on most of the factors studied. Germination boosts
nutritional content and phytase activity while decreasing phytic acid content. Germination can be a
simple method to reduce phytic acid in quinoa and may also improve the nutritional quality of this
pseudocereal with the potential for use in functional foods and vegetarian diets.

Citation: Maldonado-Alvarado, P.;

Keywords: Chenopodium quinoa; germination; phytic acid; phytase activity
Pavón-Vargas, D.J.; Abarca-Robles, J.;
Valencia-Chamorro, S.; Haros, C.M.
Effect of Germination on the
Nutritional Properties, Phytic Acid
Content, and Phytase Activity of
1. Introduction
Quinoa (Chenopodium quinoa Willd). Quinoa (Chenopodium quinoa Willd), a pseudocereal, has been cultivated in the Andean
Foods 2023, 12, 389. https://doi.org/ region for thousands of years [1]. From a nutritional point of view, quinoa is a natural
10.3390/foods12020389 source of vegetable protein with high nutritional value due to its higher proportion of
Academic Editors: Marina Carcea,
essential amino acids than in cereals [2,3]. Quinoa also has a high content of minerals,
Manuel Gomez and Eliana Pereira
such as calcium, iron, zinc, and phosphorus [3,4]. In recent times, interest in this product
has increased. It has become a popular raw material in the diet of vegetarians and people
Received: 26 October 2022 with intolerances or allergies to cereals. Researchers have shown particular interest in this
Revised: 3 January 2023 pseudocereal as a potential ingredient in gluten-free food formulations [5,6].
Accepted: 6 January 2023 Quinoa, however, has some anti-nutritional factors, especially saponins and phytic acid.
Published: 13 January 2023
Phytic acid can bind divalent minerals, making them unavailable for normal metabolism.
As a consequence, phytic acid reduces the nutritional value of quinoa [7,8]. In humans, the
consumption of 5 to 10 mg per day of phytic acid can cause a reduction in iron solubility of 50%,
possibly leading to deficiencies of the mineral [9].
Copyright: © 2023 by the authors.
Licensee MDPI, Basel, Switzerland.
The phytase enzyme catalyses the hydrolysis of phytic acid, thus releasing inorganic
This article is an open access article
phosphate from the seeds. The enzymatic activity of endogenous phytase in quinoa de-
distributed under the terms and creases with soaking, but, conversely, the enzymatic activity increases during germination
conditions of the Creative Commons due to the gradual dephosphorylation of the phytic acid complexes [10]. Germination
Attribution (CC BY) license (https:// not only reduces the amount of some anti-nutritional factors, such as phytic acid, but it
creativecommons.org/licenses/by/ also increases the amount of some nutrients, especially minerals and antioxidant com-
4.0/). pounds [7,11].

Foods 2023, 12, 389. https://doi.org/10.3390/foods12020389 https://www.mdpi.com/journal/foods

Foods 2023, 12, 389 2 of 14

Germination is an important method to increase the nutritional and functional value of

seeds. During germination, several enzymes are activated, improving protein digestibility
and mineral bioavailability [12]. At present, there is a growing interest in the consumption
of seed sprouts, alone or as a raw material for other products, because they have a higher
nutritional value and are related to a healthier lifestyle [7].
The effect of germination has been studied in cereals and pseudocereals [13,14]. Re-
cently, Pilco-Quesada et al. [14] studied the effects of germination and kilning on the
phenolic compounds and nutritional properties of quinoa. They found that germination for
72 h resulted in a significant increase in the total content of phenolics and may improve the
nutritional properties of the pseudocereal. However, even though there is some information
about quinoa nutrients and anti-nutritional factors, previous studies have focused more
on saponins [15], and there is no study focused on the phytic acid content and phytase
activity of quinoa during germination and refrigerated storage. Thus, this research aimed
at studying the effect of desaponification (saponin removal), soaking, germination, and
refrigerated storage on the proximate composition, mineral content, phytic acid content,
and phytase activity of three quinoa varieties.

2. Materials and Methods

2.1. Materials
2.1.1. Raw Materials
Three varieties of quinoa were used for the experiments. White quinoa was sup-
plied by INIAP (Instituto Nacional de Investigaciones Agropecuarias), Quito, Ecuador.
Black quinoa and red quinoa harvested in Cotopaxi Province were supplied by Inca’s
Treasure (Latacunga, Ecuador). The samples were stored at room temperature and in a
dark environment to prevent light exposure.

2.1.2. Desaponification, Germination, and Sampling Procedure

A wet method was used to remove the saponins in the seeds, which involved washing
the quinoa with distilled water. The seeds were washed four times within a water–seed
ratio of 2:1, with constant stirring at 100 rpm for 40 s. To verify the absence of saponins, a
3 g sample was shaken in a test tube together with 5 mL of distilled water. The formation
or absence of foam after rinsing allowed us to determine if an additional wash should be
carried out.
Quinoa seeds, previously desaponified, were disinfected with a Kilol® solution (1 mL
ascorbic acid, 0.475 mL citric acid, 0.47 mL lactic acid, water q.s.p. 100 mL–5 mL/L) (Chemie
Ecuador, Guayaquil, Ecuador) prior to the germination process. The soaking and sprouting
processes were carried out following the method described by D’Ambrosio et al. [12]. The
seeds were soaked for 6 h in distilled water (water–seed ratio 2:1). Subsequently, the
soaking water was drained, and the seeds were placed in trays, covered with humidified
absorbent paper, inside a seed germinator (Mangelsdorf Germination Chamber, TE-406,
Piracicaba, Brazil) for 7 days at a temperature of 20 ◦ C and relative humidity (RH) of 100%.
The sampling was carried out 5 times during germination and storage. First, after
soaking for 6 h (0.25 days), then at 4 and 7 days of germination in the chamber, and finally
at 7 days of germination plus 4 and 7 days of refrigerated storage (days 11 and 14). In
this sense, to evaluate the effect of refrigerated storage, after the 7th day of germination,
sprouts of the three varieties were packaged in polyethylene terephthalate (PET) perforated
boxes and stored in a refrigerator at a temperature of 4 ◦ C and 75% RH for seven more
days. Samples were taken at 7 plus 4 (11th) and 7 (14th) days of refrigerated storage for
subsequent lyophilisation and storage as stated previously.
After 6 h of soaking (0.25 days), and at 4 and 7 days of germination, 35 g of each
variety were sampled and frozen at −20 ◦ C. The frozen samples were then freeze-dried in
a lyophilizer (Stokes, 264 sq. ft., NC, USA), and then the samples were collected and stored
in desiccators at room temperature until the chemical composition analysis was carried out.
The whole procedure is shown in Scheme 1.
 After 6 h of soaking (0.25 days), and at 4 and 7 days of germination, 35 g of each
variety were sampled and frozen at −20 °C. The frozen samples were then freeze-dried in
Foods 2023, 12, 389 a lyophilizer (Stokes, 264 sq. ft., NC, USA), and then the samples were collected and stored 3 of 14
in desiccators at room temperature until the chemical composition analysis was carried
out.

The whole procedure is shown in Scheme 1.

Scheme 1. Desaponification, germination, and sampling procedure.

2.2. Methods
The chemical composition of the three varieties of quinoa (white, red, and black) was
monitored before soaking (day 0), after soaking (0.25 days), at 4 and 7 days of germination,
and at 4 and 7 days of refrigerated storage, to simulate the shelf-life of the sprouts.

2.2.1. Proximate Analysis

The total lipid, fibre, ash, and moisture contents of the samples were determined
according to AOAC official methods: 945.16, 985.29, 923.03, and 925.10, respectively [16].
Fibre was assessed using the test protocol adapted from the Megazyme® Total Dietary
Fibre Kit (Megazyme Ltd., Bray, Ireland). The protein content was analysed using the rapid
nitrogen determination method according to
Dumas with a nitrogen and protein analyser (Elementar, Langenselbold, Germany).
Carbohydrates were calculated using the other measurements by difference. All the analy-
ses were performed three times.

2.2.2. Mineral Content

The samples were analysed using atomic absorption spectrometry to determine the
concentration of minerals. The metal analysis was completed using a flame absorption spec-
Foods 2023, 12, 389 4 of 14

trometer at the Analysis of Soils, Plants and Water Service of the Institute of Agricultural
Sciences, Madrid (Spain). to determine the calcium, zinc, and iron concentrations following
the methodology proposed by Tazrart et al. [17]. Digestion of the samples was carried out
using a microwave (MARS, Charlotte, NC, USA) with 4 mL of nitric acid (0.88 m/v) and
1 mL of hydrogen peroxide (33.3 m/v) at 180 ◦ C. Nitric acid (≥99% purity) and hydrogen
peroxide (≥99% purity) were obtained from Merck KGaA (Darmstadt, Germany). Six sam-
ples of each variety and time were run independently. Results were reported in milligrams
(mg) per 100 g of sample.

2.2.3. Phytic Acid Content

Quantification of the phytic acid content was measured according to the method
proposed by Reason et al. [18] with a Phytic Acid Assay Kit by Megazymes® . Double
enzymatic digestion was used to release inorganic phosphorus (Pi) from the samples. To
produce a quantifiable value using UV-Vis spectroscopy, Pi was reacted with ammonium
molybdate. The content of free and total inorganic phosphorus was measured as absorbance
of molybdenum blue at a wavelength of 655 nm using a UV-Vis spectrometer (BMG Labtech,
SPECTROstar Nano, Ortenberg, Germany). To quantify phytic acid in each sample a
standard curve was prepared with dilutions of standard phosphorus. The content of free
and total phosphorus was obtained from each sample, and the difference between these
two values was the binding phosphorus. For each 0.282 g of binding phosphorus, there is
100 g of phytic acid.

2.2.4. Phytase Activity

Potassium phytate was used as a source of phosphorus. Enzyme extracts of the sam-
ples were prepared following the methodology used by Luo et al. [19], which was modified
by García-Mantrana et al. [20] for use on microplates. The absorbance of the reaction
product was measured at 400 nm using a UV-Vis Spectrophotometer (BMG Labtech, SPEC-
TROstar Nano, Ortenberg, Germany). A standard curve was prepared with a dipotassium
phosphate stock solution. Potassium phytate (≥95% purity) and dipotassium phosphate
(≥95% purity) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Phytase
enzymatic activity was measured in phytase units (U), defined as the amount of enzyme
capable of releasing 1 microgram (µg) of inorganic phosphorus per minute at a pH of 5.6
and a temperature of 50 ◦ C.

2.3. Statistical Analysis

Statistical analyses of significant differences between the raw material and the de-
saponification and soaking processes were carried out for all the response variables. The
results were expressed as mean value ± standard deviation (SD). The statistical analysis
was performed with STATGRAPHICS Centurion XV (Statpoint Technologies Inc., War-
renton, VA, USA) software. One-way ANOVA was performed to evaluate the statistical
significance of differences. At each storage moment, the difference of variables among
the different quinoa varieties was tested with a post-hoc analysis using Fisher’s test (LSD)
(p ≤ 0.05).

3. Results and Discussion

3.1. Chemical Composition of the Raw Material
The chemical composition of the raw quinoa samples is shown in Table 1. The moisture,
lipid, protein, ash, and fibre contents are shown as percentages on a dry basis. There were
significant differences (p ≤ 0.05) between the three varieties: white quinoa (WQ), red quinoa
(RQ), and black quinoa (BQ) in moisture content, lipids, and ash.
Foods 2023, 12, 389 5 of 14

Table 1. Chemical composition of the raw quinoa samples (d.b.).

White Quinoa Red Quinoa Black Quinoa

c a
Moisture (%) 13.26 ± 0.04 12.44 ± 0.01 12.55 ± 0.04 b
Lipids (%) 7.92 ± 0.06 c 7.56 ± 0.13 b 7.02 ± 0.13 a
Crude protein (%) 19.26 ± 0.95 b 15.90 ± 0.12 a 16.06 ± 0.04 a
Ash (%) 2.27 ± 0.02 a 3.25 ± 0.04 b 4.27 ± 0.08 c
Fibre (%) 7.02 ± 3.68 a 19.55 ± 2.43 b 7.68 ± 1.24 a
Carbohydrates (%) 50.27 41.3 53.42
Calcium (Ca) (mg/100 g) * 64.88 ± 0.84 a 76.76 ± 4.87 b 74.72 ± 8.49 b
Zinc (Zn) (mg/100 g) * 3.83 ± 0.08 b 1.77 ± 0.16 a 1.73 ± 0.25 a
Iron (Fe) (mg/100 g) * 4.45 ± 0.36 a 4.81 ± 0.25 a 4.68 ± 0.48 a
Phytic acid (g/100 g) 1.07 ± 0.01 b 1.03 ± 0.03 a 1.22 ± 0.01 c
Phytase activity (U) 41.56 ± 0.99 c 37.01 ± 0.34 a 40.01 ± 0.39 b
Mean ± SD ( n = 3, n = 6). Results with different superscript letters within the same row are significantly different
at p ≤ 0.05. The “*” means that the umber of determination of Ca, Fe and Zn were 6. [Mean ± SD, n = 3, (* n = 6)].

In all varieties, the protein content ranged from 15.90% to 19.26%, which was in
accordance with Valencia-Chamorro, who reported protein contents for quinoa between
8.0% and 22.0% [1,21]. Other authors have reported lower and higher crude protein values
than those of the present study [7,22]. WQ protein content was significantly higher (18%)
than that in the RQ and BQ samples. The ash content of the studied samples ranged from
2.27% to 4.27%, and the lipid content was around 7.5%, which is comparable to values in
previous studies [14]. Although the fibre content values for the black and white varieties
did not show significant differences and were in the range found in the literature, the fibre
content value in the red variety was around 19.55%, which was much higher than the range
reported for crude fibre in quinoa (2.5–3.9%) [1,14]. The reason for the high variability
in the proximate analysis among the quinoa samples may be due to the variety of each
seed. It is known that quinoa varieties and the growing environment can affect protein
content [2,23].
As can be seen in Table 1, the mineral content of samples in this investigation showed
significant differences (p ≤ 0.05) for calcium and zinc. The calcium content in RQ and BQ
was 76.76 and 74.72 mg/100 g, respectively, which was significantly higher (14.3%) than
the calcium content in WQ (64.88 mg/100 g). Conversely, the zinc content in WQ was
significantly higher (54.3%) than the zinc content in the RQ and BQ varieties. The mineral
content values obtained in this study were similar to those described in the literature [4,24].
It is important to note that the presented values for the chemical composition cor-
respond to quinoa seeds before desaponification and without scarification. The results
obtained do not consider losses from the desaponification process, e.g., Repo-Carrasco-
Valencia and Astuhuaman Serna [22] reported losses between 0.71 and 2.95 g/100 g of
protein after desaponification of quinoa seeds.

3.2. Phytic Acid Content and Phytase Activity of the Raw Material
The phytic acid content of BQ (1.22 g/100 g) was significantly higher than that of
WQ (1.07 g/100 g) and RQ (1.03 g/100 g) (Table 1). It has been reported that, for different
varieties of quinoa, the average value of the phytic acid content is 1.18 g/100 g [1]. Higher
values up to 1.94 g/100 g [25] and lower values (0.97 g/100 g) have been reported for the
white quinoa variety [7,25]. The phytic acid content in quinoa, in general, presents higher
values than those of other cereals and seeds [10]. In quinoa, phytic acid is not only present
in the outer layers of the seed, as in other cereals, but is also evenly distributed in the
endosperm [26]. A high phytic acid content can have a negative effect on nutrition and
diet [9]. A low phytic acid content is preferred.
Phytase activity was measured in phytase units (U). As can be seen in Table 1, RQ pre-
sented the lowest phytase activity value of 37.01 ± 0.34 U, BQ showed a value of 40.01 ± 0.39 U,
and the highest phytase activity was found in WQ at a value of 41.56 ± 0.99 U. There were
significant differences (p ≤ 0.05) between the three varieties.
 Phytase activity was measured in phytase units (U). As can be seen in Table 1, RQ
presented the lowest phytase activity value of 37.01 ± 0.34 U, BQ showed a value of 40.01
Foods 2023, 12, 389
± 0.39 U, and the highest phytase activity was found in WQ at a value of 41.56 ± 0.99 6U.
of 14
There were significant differences (p ≤ 0.05) between the three varieties.

3.3. Effect of Germination and Refrigerated Storage on the Chemical Composition

3.3. Effect of Germination and Refrigerated Storage on the Chemical Composition
3.3.1. Protein Content
3.3.1. Protein Content
The protein content of the studied samples decreased after soaking but increased
The protein content of the studied samples decreased after soaking but increased
during germination. Figure 1 shows the change in protein content for the three varieties
during germination. Figure 1 shows the change in protein content for the three varieties
of quinoa during soaking, germination, and refrigerated storage. As can be seen, after the
of quinoa during soaking, germination, and refrigerated storage. As can be seen, after the
desaponification
desaponification and soaking
and processes
soaking processes(0.25 days),
(0.25 the
days), total
the protein
total content
protein contentininallallthe
theva-
vari-
rieties had reduced from the initial value. WQ showed the most significant reduction
eties had reduced from the initial value. WQ showed the most significant reduction (59%)
(59%) in protein
in protein compared
compared to the
to the rawraw materials,
materials, while
while in RQ
in RQ andand
BQ,BQ,
the the protein
protein concen-
concentration
tration was reduced by 40% and 14%, respectively.
was reduced by 40% and 14%, respectively.

Figure
Figure 1. Protein
1. Protein content
content variation
variation for for white
white (WQ),
(WQ), redred (RQ),
(RQ), andand black
black (BQ)(BQ) quinoa,
quinoa, during
during soaking
soaking
(0.25 days), germination (4 and 7 days ◦
(0.25 days), germination (4 and 7 days at 20at°C
20andC 100%
and 100%
RH), RH), and refrigerated
and refrigerated storage
storage (1114and
(11 and
days14 at 4 °C
days 4 ◦ C 75%
at and RH).RH).
and 75% ErrorError
barsbars
indicate standard
indicate deviation
standard (n =(n3).= Results
deviation with
3). Results different
with different
letters areare
letters significantly different
significantly at pat≤p0.05.
different ≤ 0.05.

According
According to Repo-Carrasco-Valencia
to Repo-Carrasco-Valencia andand Astuhuaman
Astuhuaman Serna Serna
[22], [22], the desaponifi-
the desaponifica-
tioncation
process process
causescauses a decrease
a decrease in protein
in protein content.content. They reported
They reported a reduction
a reduction of up to of 7%
up to
7% protein
in the in the protein
contentcontent
of quinoaof quinoa after desaponification.
after desaponification. The soaking
The soaking processprocess
was notwas con-not
considered
sidered in theirinstudy,
their study,
whichwhich
could could
explain explain
the more the significant
more significant percentage
percentage of proteinof protein
re-
reduction
duction in the inpresent
the present
study.study.
The The
main main protein
protein present
present in quinoa
in quinoa is globulin
is globulin [27],[27], which,
which,
since
since it isit soluble
is soluble in water,
in water, could
could be be leached
leached as as a result
a result of of
thethe soaking
soaking processes
processes together
together
with other soluble proteins
with other soluble proteins and molecules. and molecules.
Germination
Germination hadhada asignificant effect (p(p≤≤0.05)
significant effect 0.05)ononthethe
protein content
protein of quinoa
content sprouts
of quinoa
compared
sprouts comparedto the to
values after soaking.
the values There was
after soaking. There an was
increasing trend in trend
an increasing the three
in thevarieties
three of
quinoaof
varieties during
quinoa germination. On day 4, WQ
during germination. On and
day BQ increased
4, WQ and BQ their protein concentration
increased their protein by
1.29 and 1.23bytimes,
concentration respectively,
1.29 and 1.23 times,compared to thecompared
respectively, protein concentration
to the protein after the desaponi-
concentration
fication and soaking processes. However, RQ required
after the desaponification and soaking processes. However, RQ required a longera longer germination timegermi-
(7 days)
nation time (7 days) to achieve a significant difference compared to the protein concentra-An
to achieve a significant difference compared to the protein concentration after soaking.
tionincrease in protein
after soaking. Ancontent
increasecaninbeprotein
also attributed
content can to weight
be alsoloss during respiration
attributed to weight loss due to
the carbohydrates and lipids consumption pathways, as absolute
during respiration due to the carbohydrates and lipids consumption pathways, as abso- protein content cannot
lutechange
protein [14]. In thiscannot
content study,change
a reduction
[14]. in
In the
thislipid
study,and carbohydrate
a reduction content
in the lipid was
and observed
carbo-
for all the samples. The quinoa variety was also a factor that
hydrate content was observed for all the samples. The quinoa variety was also a factoraffected protein concentration,
and there were significant differences between the varieties. At the end of the study, RQ
reached a significantly higher level of protein content than WQ and BQ. In contrast to the
results found, Chaparro et al. [28] showed that there were no significant differences for the
‘Valle del Cauca’ quinoa variety after the germination process.
 that affected protein concentration, and there were significant differences between the va-
rieties. At the end of the study, RQ reached a significantly higher level of protein content
Foods 2023, 12, 389
than WQ and BQ. In contrast to the results found, Chaparro et al. [28] showed that there 7 of 14
were no significant differences for the ‘Valle del Cauca’ quinoa variety after the germina-
tion process.
Following the plus 7 days of refrigerated storage (4 °C and 75% RH) after the germi-
◦ C and 75% RH) after the ger-
Following
nation process, no the plus 7 days
differences wereofshown
refrigerated
in any storage
sample (4
until day 14. The protein con-
mination process, no differences were shown in any sample until day 14. The protein
centration in the sprouts remained at the levels reached during germination.
concentration in the sprouts remained at the levels reached during germination.
3.3.2. Ash Content
3.3.2. Ash Content
TheThe
variation in ash
variation content
in ash of the
content quinoa
of the samples
quinoa during
samples soaking,
during germination,
soaking, germination,andand
storage is given in Figure 2. Ash contents of the quinoa sprouts at the end of the
storage is given in Figure 2. Ash contents of the quinoa sprouts at the end of the storage storage
changed
changed between
between 2.11 and
2.11 and4.24%. The
4.24%. germination
The germination process increased
process increasedthe
thevalues
valuesbetween
between 9
9 and 18% relative to the values after soaking (1.78–3.88%). The final
and 18% relative to the values after soaking (1.78–3.88%). The final values values were close
were to to
close
those reported for the initial raw material (0 days), which were between 2.27 and
those reported for the initial raw material (0 days), which were between 2.27 and 4.27%. 4.27%.

Figure
Figure2. Ash content
2. Ash variation
content for for
variation white (WQ),
white redred
(WQ), (RQ), andand
(RQ), black (BQ)
black quinoa
(BQ) during
quinoa soaking
during soaking
(0.25 days),
(0.25 germination
days), (4 and
germination 7 days
(4 and at 20at°C20and
7 days ◦ C 100% RH), RH),
and 100% and refrigerated storage
and refrigerated (11 and
storage (1114and
days14 at
days4 °C
at and
4 ◦ C75%
and RH). ErrorError
75% RH). bars bars
indicate standard
indicate deviation
standard (n =(n3).= Results
deviation with
3). Results different
with different
letters are significantly different at p ≤ 0.05.
letters are significantly different at p ≤ 0.05.

Germination
Germination significantly
significantly affected
affected(p ≤(p0.05)
≤ 0.05)the the
ashash
content
contentpresent
presentin the sprouts
in the sprouts
after soaking.
after soaking. In the white
In the whitevariety, after
variety, thethe
after desaponification
desaponification andandsoaking
soakingprocesses,
processes, thethe
ashashcontent
contentdecreased
decreased by by
22% 22%compared
compared to the raw
to the rawmaterial,
material,andandthen, thethe
then, ashash
content
content
significantly
significantly increased
increased (p ≤(p0.05) untiluntil
≤ 0.05) day 7day of germination. The same
7 of germination. The behaviour was ob-
same behaviour was
served in the red variety but without a statistical difference between
observed in the red variety but without a statistical difference between the fourth the fourth and sev-and
enth days ofdays
seventh germination. In these In
of germination. two varieties,
these there were
two varieties, no changes
there were noinchanges
the ash content
in the ash
during
contentrefrigerated storage, and
during refrigerated the values
storage, and thereached
valuesonreached
day 7 of ongermination were main-
day 7 of germination were
tained. For the black
maintained. For thevariety
black (BQ),
varietythe effect
(BQ), theofeffect
germination and refrigerated
of germination storagestorage
and refrigerated had
a different trend than
had a different trendobserved in the previous
than observed varieties.
in the previous In BQ,In
varieties. the
BQ,ash
thecontent present
ash content in
present
theinrawthe material
raw material decreased by 9%
decreased byafter
9% aftersoaking;
soaking;subsequently,
subsequently, the the
value progressively
value progressively
increased
increased during
during germination
germination and then
and thenduring
during refrigerated
refrigeratedstorage
storageuntil day
until 14.14.
day
TheTheinitial reduction
initial reduction of ash content
of ash content during
during soaking cancan
soaking be be
explained
explained by by
thethe
losslossof of
minerals
minerals duedue to to
lixiviation
lixiviation [14].
[14].Meanwhile,
Meanwhile,the theincrease
increaseininash
ashcontent
content during germination
germina-
tionmay
maybeberelated
relatedtotothetheconversion
conversionofofcarbohydrates
carbohydratestotocarboncarbondioxide
dioxideduring
duringrespiration,
respira-
therefore,
tion, therefore, increasing
increasing thethe
percentage
percentage of the ashash
of the on on
a dry basis.
a dry basis.These
Thesedata corroborate
data corroborate those
found
those in other
found in otherstudies for quinoa
studies and quinoa
for quinoa and quinoaflour,flour,
whichwhich
showed significant
showed increases
significant in- in
ash content
creases after germination
in ash content of 33%of
after germination and
33% 46%,
andrespectively [7,29].[7,29].
46%, respectively Nevertheless, another
Nevertheless,
study indicated a reduction in ash content in sprouted quinoa compared to the raw material,
which was explained by the use of minerals as coenzymes during the bioconversion of
carbohydrates [14].
There was an increase in ash content in the black variety due to refrigeration. Refrig-
erated storage should preserve the original composition, as in the case of ash. Metabolic
activity during refrigeration, including respiration, consumes water and other products,
 another study indicated a reduction in ash content in sprouted quinoa compared to the
raw material, which was explained by the use of minerals as coenzymes during the bio-
conversion of carbohydrates [14].
Foods 2023, 12, 389
There was an increase in ash content in the black variety due to refrigeration. Refrig-
8 of 14
erated storage should preserve the original composition, as in the case of ash. Metabolic
activity during refrigeration, including respiration, consumes water and other products,
such as carbohydrates or lipids. This can be a form in which the ash increases in percent-
such as carbohydrates or lipids. This can be a form in which the ash increases in percentage
age by reducing the content of other components. Furthermore, in this study, the relative
by reducing the content of other components. Furthermore, in this study, the relative
humidity in refrigeration was lower than in the germination process. Thus, the water of
humidity in refrigeration was lower than in the germination process. Thus, the water of
the product could be lost more easily in the sprouts and, therefore, a higher ash content
the product could be lost more easily in the sprouts and, therefore, a higher ash content
during refrigerated storage could be achieved. Quinoa has high genetic variability, and
during refrigerated storage could be achieved. Quinoa has high genetic variability, and the
the different increments in the amount of ash present in the samples during germination
different increments in the amount of ash present in the samples during germination and
and storage could be linked to it [30].
storage could be linked to it [30].
3.3.3. Fibre
3.3.3. Content
Fibre Content
Figure 3 shows
Figure 3 shows thethe
effects of the
effects treatments
of the treatmentson on
thethe
fibre content
fibre of the
content three
of the quinoa
three quinoa
varieties. Desaponification and soaking caused a significant reduction in
varieties. Desaponification and soaking caused a significant reduction in fibre content fibre content of
between 28 and2878%.
of between and The
78%.loss
The of loss
the perianth during during
of the perianth desaponification and soaking
desaponification may
and soaking
explain this decrease.
may explain The perianth
this decrease. in the quinoa
The perianth in theseeds
quinoacontains
seeds insoluble fibre that fibre
contains insoluble can bethat
lostcan
during soaking [31]. The fibre content values after soaking, as well as
be lost during soaking [31]. The fibre content values after soaking, as well as duringduring germi-
nation and refrigerated
germination storage,storage,
and refrigerated were within the values
were within found
the values for raw
found andand
for raw germinated
germinated
quinoa [1,14].
quinoa [1,14].

Figure
Figure 3. Fibre
3. Fibre content
content variation
variation for for
whitewhite (WQ),
(WQ), redred (RQ)
(RQ) andand black
black (BQ)(BQ) quinoa
quinoa during
during soaking
soaking
(0.25 days),
(0.25 germination
days), (4 and
germination 7 days
(4 and 7 days ◦ C 100%
at 20at°C20and RH), RH),
and 100% and refrigerated storage
and refrigerated (11 and
storage (1114and
days14 at 4 °C
days at and
4 ◦ C 75% RH).RH).
and 75% ErrorError
bars bars
indicate standard
indicate deviation
standard (n =(n3).= Results
deviation with
3). Results different
with different
letters areare
letters significantly different
significantly at pat≤p0.05.
different ≤ 0.05.

Germination
Germination significantly increased
significantly (p ≤(p0.05)
increased the the
≤ 0.05) fibre content
fibre values
content valuesforfor
thethe
three
three
varieties compared
varieties compared to values after
to values soaking
after soaking(0.25 days).
(0.25 days).TheThesynthesis of fibre
synthesis within
of fibre withinthethe
structures,
structures,such as as
such thethehypocotyl
hypocotyl andand
thethe
radicle, of of
radicle, quinoa
quinoa seeds during
seeds during germination
germination
could explain
could explainthisthis
increase [32].
increase However,
[32]. However,there was
there no no
was significant change
significant change between
between thethe
values of the
values 4th4th
of the dayday(between 5.84
(between andand
5.84 8.89%)
8.89%)andand
thethe
14th dayday
14th (between
(between5.485.48
andand
9.13%)
9.13%)
after refrigerated
after refrigerated storage, although
storage, although there were
there differences
were differences between
betweenthethe
three varieties.
three varieties.
Similar
Similar results
results were
were described
described forfor quinoa
quinoa after
after 3 days
3 days of germination,
of germination, where
where there
there was was
no no
significant
significant effect
effect of the
of the germination
germination process
process ononthethe crude
crude fibre
fibre content
content [14].
[14].

3.3.4. Lipid and Carbohydrate Content

Total lipid content in the three varieties showed a significant difference between the
sampling days. A decreasing trend during germination and refrigerated storage was
observed (Figure 4). During the autotrophic development of quinoa, seeds do not perform
photosynthesis, and lipids are most likely a source of energy during germination [32]. As for
carbohydrates, they also decrease as the germination process progresses, as carbohydrates
are used as an energy source during the germination process (Table 2). The seeds use
 d.b.
BQ 73.9 ± 5.9 f 84.4 ± 4.1 g 94.1 ± 7.3 hi 91.5 ± 7.6 gh 97.0 ± 8.2 i
WQ 4.0 ± 0.1 h 4.4 ± 0.2 i 4.6 ± 0.1 i 4.7 ± 0.0 i 4.7 ± 0.3 i
mg/100 g
Zn RQ 1.8 ± 0.0 ab 2.6 ± 0.6 fg 2.8 ± 0.3 g 2.1 ± 0.1 cd 2.2 ± 0.2 cde
d.b.
BQ 1.7 a 2.0 abc 2.1 bcd 2.2 de 2.4 ef
Foods 2023, 12, 389 WQ 4.0 ± 0.4 abde nd 5.2 ± 0.5 efg 4.9 ± 0.1 cdfg 5.0 ± 0.2 defg9 of 14
mg/100 g
Fe RQ 3.0 ± 0.1 a nd 3.4 ± 0.1 ab 3.6 ± 0.1 abc 3.3 ± 0.5 ab
d.b.
BQ 4.4 ± 0.4 bdf nd 5.7 ± 0.9 fg 5.8 ± 0.3 g 4.9 ± 0.3 bdfg
Meancarbohydrates and lipids
± SD, n = 6. Results withfor biochemical
different activities
superscript lettersdue to germination
within the same row [33].
are This reduction
significantly
different at pand
in lipid ≤ 0.05. * Carbohydrates
carbohydrate were
content wascalculated based on
also observed inthe other component
a study measurements
by Pilco-Quesada et al. [14]
by difference.
on germinated n.d.: No determined.
quinoa seeds.

Figure
Figure 4. Lipid
4. Lipid content
content variation
variation for for white
white (WQ),
(WQ), redred (RQ),
(RQ), andand black
black (BQ)
(BQ) quinoa
quinoa during
during soaking
soaking
(0.25 days),
(0.25 germination
days), (4 and
germination 7 days
(4 and 7 days ◦ C 100%
at 20at°C20and RH), RH),
and 100% and refrigerated storage
and refrigerated (11 and
storage (1114and
14 days at 4 ◦ C and 75% RH). Error bars indicate standard deviation (n = 3). Results with different
letters are significantly different at p ≤ 0.05.

Table 2. Carbohydrate and mineral content variation for white (WQ), red (RQ), and black (BQ) quinoa
during soaking (0.25 days), germination (4 and 7 days at 20 ◦ C and 100% RH), and refrigerated storage
(11 and 14 days at 4 ◦ C and 75% RH).

Desaponification Refrigeration Refrigeration

Germination 4 Germination 7
Component Units Quinoa and Soaking Storage Storage
Days Days
0.25 Days 4 Days 7 Days
WQ 71.67 66.04 65.29 64.80 64.80
Carbohydrates* g/100 g d.b. RQ 63.96 60.96 60.46 58.55 56.60
BQ 67.47 61.24 61.95 60.74 60.14
WQ 60.5 ± 2.4 bcd 66.2 ± 4.4 de 75.6 ± 5.4 f 74.4 ± 7.0 f 71.3 ± 3.9 ef
Ca mg/100 g d.b. RQ 51.6 ± 0.3 a 56.8 ± 1.1 ab 64.6 ± 1.4 cde 57.5 ± 4.0 abc 63.2 ± 6.5 bcd
BQ 73.9 ± 5.9 f 84.4 ± 4.1 g 94.1 ± 7.3 hi 91.5 ± 7.6 gh 97.0 ± 8.2 i
WQ 4.0 ± 0.1 h 4.4 ± 0.2 i 4.6 ± 0.1 i 4.7 ± 0.0 i 4.7 ± 0.3 i
Zn mg/100 g d.b. RQ 1.8 ± 0.0 ab 2.6 ± 0.6 fg 2.8 ± 0.3 g 2.1 ± 0.1 cd 2.2 ± 0.2 cde
BQ 1.7 a 2.0 abc 2.1 bcd 2.2 de 2.4 ef
WQ 4.0 ± 0.4 abde nd 5.2 ± 0.5 efg 4.9 ± 0.1 cdfg 5.0 ± 0.2 defg
Fe mg/100 g d.b. RQ 3.0 ± 0.1 a nd 3.4 ± 0.1 ab 3.6 ± 0.1 abc 3.3 ± 0.5 ab
BQ 4.4 ± 0.4 bdf nd 5.7 ± 0.9 fg 5.8 ± 0.3 g 4.9 ± 0.3 bdfg
Mean ± SD, n = 6. Results with different superscript letters within the same row are significantly different at
p ≤ 0.05. * Carbohydrates were calculated based on the other component measurements by difference. n.d.: No
determined.

3.3.5. Calcium
Germination brought about a great increase in the calcium content of the samples.
The increase ranged from 25% to 27% in the three varieties compared to the material
after desaponification and soaking, and it increased significantly for the white and black
varieties, by 17% and 26%, respectively, compared to the values of the initial raw material
(Table 2). Calcium content in the samples at the end of the germination period ranged
between 75.9 and 93.5% mg/100 g (d.b.). Quinoa sprouts have a relatively higher content
of calcium compared to raw quinoa and other cereals [1].
Similar results for the influence of quinoa seed germination on calcium concentration
were obtained by Chaparro et al. [28] who described an increase of 25% in quinoa after
Foods 2023, 12, 389 10 of 14

two days of germination. Germination, in general, has been shown to promote the avail-
ability of minerals in cereals [33]. Contrary to this, refrigerated storage did not affect the
calcium content in any of the germinated samples.
The loss of the perianth during saponification, together with the leaching resulting
from soaking, are two determinant factors in the initial reduction of calcium content during
these processes [34]. Although, the increase in calcium content during germination may
be related to the phytic acid content and phytase activity. As germination progresses, the
chelation of metals becomes reduced. Therefore, a higher content of calcium, iron, and zinc
is present in the samples.

3.3.6. Zinc
After the desaponification and soaking processes, WQ presented the highest increase
in zinc content with respect to raw materials by 5%, while in RQ, the zinc content increased
by 1.8%, and in BQ, the concentration of zinc decreased by 0.8% (Table 2).
The germination process produced a significant increase (p ≤ 0.05) in the zinc content
after 4 days of germination in all the varieties, which increased by 10, 14, and 45% for the
WQ, BQ, and RQ, respectively, when compared to the zinc contents after desaponification
and soaking. Furthermore, the zinc concentrations after seven days of germination were
higher than in the raw materials by 15, 13, and 43% for WQ, BQ, and RQ, respectively.
Although refrigerated storage did not influence the zinc content in WQ and BQ after
germination, it brought a significant decrease (p ≤ 0.05), by 25% at the 7 days plus 7 days
of refrigerated storage, in the red variety.
Luo et al. [35] showed an increase in the availability of zinc for wheat and rice seeds,
by 80 and 87%, respectively, after germination. Nevertheless, in soybeans, zinc availability
decreased by 73%. Zinc participates in the hydrolysis of reserve substances in the seeds,
and this could reduce its content during refrigeration, for example. On the other hand,
during germination, the increase in zinc content could be explained by the physiology of
the plant itself [36].

3.3.7. Iron
All the varieties had similar iron contents, ranging from 4.45 to 4.81 mg/100 g (d.b.) in
the raw material. After the desaponification and soaking processes, RQ showed a decrease
of 37% in iron content. Meanwhile, in the WQ and BQ varieties, the reduction in iron
concentration was 10 and 5.53%, respectively (Table 2). Iron concentration for the WQ
and BQ varieties increased significantly (p ≤ 0.05) with germination until the seventh day.
Nevertheless, the RQ variety did show significant differences (15%) in iron concentration
due to germination. Chaparro et al. [28] reported a significant increase (11.40%) after one
day of germination in the ‘Valle del Cauca’ quinoa variety. Moreover, the germination
process significantly increased the amount of iron in other cereals, with a 2.45% and 4.24%
increase for rice and wheat, respectively [35]. Refrigerated storage, however, did not have
any effect on the iron content of the quinoa sprouts as the values remained similar to those
found on day 7 of germination in all varieties.
Since phytic acid is one of the factors affecting mineral bioavailability, its reduction
during germination would enhance the mineral content [33]. Thus, germination generally
increased the availability of minerals in the quinoa samples. Another factor that influenced
the mineral content was leaching during soaking. Not only do the germination and leaching
processes influence the concentration of calcium, zinc, and iron, but also the variety. The
nutritional composition of quinoa greatly varies according to some factors, including the
genotype of quinoa [30].
 increased the availability of minerals in the quinoa samples. Another factor that influ-
enced the mineral content was leaching during soaking. Not only do the germination and
leaching processes influence the concentration of calcium, zinc, and iron, but also the va-
riety. The nutritional composition of quinoa greatly varies according to some factors, in-
Foods 2023, 12, 389 cluding the genotype of quinoa [30]. 11 of 14

3.4. Effect of Germination and Refrigerated Storage on the Phytic Acid Content and Phytase
Activity
3.4. Effect of Germination and Refrigerated Storage on the Phytic Acid Content and
3.4.1. EffectActivity
Phytase on the Phytic Acid Content
TheEffect
3.4.1. effect on
of germination
the Phytic Acid andContent
refrigerated storage on phytic acid content for the three
quinoa varieties is presented in Figure 5. A statistically
The effect of germination and refrigerated storagesignificant
on phyticreduction in phytic
acid content acid
for the three
was observed due to desaponification and soaking. A second significant
quinoa varieties is presented in Figure 5. A statistically significant reduction in phyticreduction wasacid
observed for all treatments
was observed at the beginning
due to desaponification andofsoaking.
germination until significant
A second day 4. Afterwards,
reductionthewas
content of phytic
observed for allacid on the last
treatments day
at the of refrigerated
beginning storage was
of germination untilalso
daysignificantly
4. Afterwards, de-the
creased
contentasofcompared
phytic acid toondaythe11 ofday
last refrigeration and storage
of refrigerated the values
was in germination.
also significantlyThe de-
decreased
crease in phytic acid
as compared to daycontent
11 ofinrefrigeration
pulses and cereals
and the during
valuesgermination has been
in germination. Thefrequently
decrease in
reported in the
phytic acid literature
content [10]. The
in pulses phytateduring
and cereals reduction after thehas
germination whole
beenprocess wasreported
frequently 22%,
41%, and 79% in WQ, RQ, and BQ, respectively. The reduction was
in the literature [10]. The phytate reduction after the whole process was 22%, 41%,very effective in the
and
BQ79%
where the molar
in WQ, RQ, andratioBQ,
of Ca and Zn were
respectively. Thelower than the
reduction wasthreshold valuein
very effective forthe
inhibition
BQ where
of mineral
the molar bioavailability
ratio of Ca and in Zn
humans [37]. than the threshold value for inhibition of mineral
were lower
bioavailability in humans [37].

Figure
Figure 5. Effect
5. Effect of germination
of germination andand refrigerated
refrigerated storage
storage ononthe
thephytic
phyticacid
acidcontent
contentininthree
threevarieties
varieties of
of quinoa: white (WQ), red (RQ), and black (BQ). Error bars indicate standard deviation 3).
quinoa: white (WQ), red (RQ), and black (BQ). Error bars indicate standard deviation (n = (n =Results
3).
with with
Results different lettersletters
different are significantly different
are significantly at p ≤at0.05.
different p ≤ 0.05.

The phytic acid content in the three quinoa varieties after the soaking process de-
The phytic acid content in the three quinoa varieties after the soaking process de-
creased. This behaviour could be inferred by the leaching during the soaking of the
creased. This behaviour could be inferred by the leaching during the soaking of the seeds
seeds [10]. During germination on day 4, WQ and BQ presented phytic acid values of
[10]. During germination on day 4, WQ and BQ presented phytic acid values of 0.72 and
0.72 and 0.32 g/100 g (d.b.), respectively; these values were lower (32.7% and 73.7%, re-
0.32 g/100 g (d.b.), respectively; these values were lower (32.7% and 73.7%, respectively)
spectively) than the content of phytic acid reported for the raw materials and seeds after
than the content of phytic acid reported for the raw materials and seeds after soaking.
soaking. However, RQ took three more days of germination to reach the minimum value,
However, RQ took three more days of germination to reach the minimum value, with a
with a phytic acid content value of 0.63 g/100 g (d.b.), which was 67.9% lower than the raw
material. The most significant reduction in phytic acid was found in black quinoa with a
decrease of 73.54% compared to the initial material.
It has been shown that the soaking and germination processes had a significant effect
on the decrease of phytic acid content in beans [38]. The positive effect of increasing the
content of some minerals may be also attributed to the decrease in phytic acid content as
a result of germination. The phytate complex is known as a chelating agent that reduces
mineral contents [19]. It has been reported that during germination, cereals and legumes
produce synthesis and activation of endogenous phytases which reduces phytate levels [39].

3.4.2. Effect on Phytase Activity

Figure 6 shows the effect of germination and refrigerated storage on the phytase
activity of the three quinoa varieties. The phytase activity in the quinoa samples was
significantly increased as a result of germination. The percentage increase on day 7 was
 produce synthesis and activation of endogenous phytases which reduces phytate levels
[39].

3.4.2. Effect on Phytase Activity

Figure 6 shows the effect of germination and refrigerated storage on the phytase ac-
Foods 2023, 12, 389 12 of 14
tivity of the three quinoa varieties. The phytase activity in the quinoa samples was signif-
icantly increased as a result of germination. The percentage increase on day 7 was found
to be between 64.10 and 149.00% compared to the reduced values after soaking. Accord-
found
ingly, to begermination,
during between 64.10theand 149.00%
phytate compared
levels decreasedto the reduced
while values activity
the phytase after soaking.
in-
Accordingly, during germination, the phytate levels decreased while the phytase
creased. During soaking (0.25 days), the phytase activity decreased compared to the initial activity
increased.
material During
by 31.97, soaking
33.85, (0.25 for
and 37.01% days),
BQ, the
RQ,phytase
and WQ, activity decreased
respectively. The compared to the
phytase activ-
initial material by 31.97, 33.85, and 37.01% for BQ, RQ, and WQ, respectively.
ity in seeds was reduced during soaking, and this decrease could be explained by phytase The phytase
activity
leaching in seeds was reduced during soaking, and this decrease could be explained by
[10].
phytase leaching [10].

Figure
Figure 6. Effect
6. Effect of germination
of germination and refrigerated
and refrigerated storagestorage on phytase
on phytase enzymatic
enzymatic activity activity
in three in three
vari-
varieties
eties of quinoa:
of quinoa: whitered
white (WQ), (WQ), redand
(RQ), (RQ), and(BQ).
black blackError
(BQ).bars
Error bars indicate
indicate standardstandard deviation
deviation (n =
3). Results with different
(n = 3). Results letters are
with different significantly
letters different
are significantly at p ≤ 0.05.
at p ≤ 0.05.
different

In the
In the WQ WQandand
RQRQ varieties,
varieties, an an increasing
increasing trend
trend in in phytase
phytase activity
activity was was observed
observed
during germination until day seven, whereas, for the black variety, the
during germination until day seven, whereas, for the black variety, the increase was increase was shown
until the fourth day. WQ had the highest value for phytase activity
shown until the fourth day. WQ had the highest value for phytase activity of the threeof the three varieties
by theby
varieties 7ththeday
7thof
daystorage, withwith
of storage, a value of 68.65
a value of 68.65U. U.
According
According totoSung
Sungetetal.
al.[40],
[40], an
an increase in phytase activity can be explained by the synthesis of new enzymes duringthe
increase in phytase activity can be explained by the synthesis of new enzymes during
thegermination
germinationprocess.
process.
Refrigeration hadhad
Refrigeration a negative
a negative effect
effect on on
thethe phytase
phytase activity.
activity. TheThe phytase
phytase activity
activity waswas
reduced during refrigerated
reduced during refrigerated storage until day 14, which can possibly be explained by thethe
storage until day 14, which can possibly be explained by
low temperature (4 ◦ C), which was far from optimal for this enzyme [41]. The decrease in
low temperature (4 °C), which was far from optimal for this enzyme [41]. The decrease in
thethe white
white variety
variety was
was upuptoto 50.98UUatat77days
50.98 daysofofrefrigerated
refrigerated storage.
storage. For
For the
the RQ
RQ and
andBQ,BQ,the
enzymatic activity had a lower value, and no significant differences were found between
the two samples after 7 days of refrigerated storage. The reduction occurs from day 7 of
germination. However, there were no significant differences between days 11 and 14 of
refrigerated storage. According to Spier et al. [41], who studied the effect of refrigerated
storage on phytase activity, a temperature reduction up to 4 ◦ C causes a decrease in phytase
activity in a range that fluctuates between 36.2 and 40.0%. This reduction may also be due
to the degradation of phytase by the protease enzymes [42].

4. Conclusions
In the present work, it was found that the white quinoa variety had the highest
protein, lipid, and zinc content, while the red variety had the highest fibre content. In
general, desaponification and soaking reduced the nutritional content of the samples in
the three varieties. However, germination boosts nutritional content and phytase activity
while decreasing phytic acid content. It is concluded that the germination process can be a
simple method to increase phytase activity and decrease the phytic acid content, as well
as improve the nutritional values of quinoa. In this study, it was found that germination
caused an increase in fibre and protein content, along with an increase in the percentage of
Foods 2023, 12, 389 13 of 14

calcium, zinc, and iron. After 7 days of germination, the most increased calcium and iron
contents were found in the white variety. Meanwhile, refrigerated storage did not produce
any significant change in the nutritional content. The most increased enzymatic activity
was found in white quinoa after 7 days of germination, while the largest reduction in phytic
acid content was presented in the black variety after refrigeration. A maximum activity
value for the phytase enzymes was observed on day 7 of germination, and then a decrease
was observed during refrigerated storage. The germination process has the potential to be
an easy method to create a functional product for food production and vegetarian diets.

Author Contributions: Conceptualization, P.M.-A., S.V.-C. and C.M.H.; methodology, P.M.-A., S.V.-C.
and C.M.H.; formal analysis, J.A.-R. and D.J.P.-V.; investigation, J.A.-R.; resources, P.M.-A., S.V.-C.
and C.M.H.; writing—original draft preparation, D.J.P.-V. and J.A.-R.; writing—review and editing,
D.J.P.-V., P.M.-A., S.V.-C. and C.M.H.; supervision, S.V.-C. and C.M.H.; project administration, P.M.-
A. and C.M.H.; funding acquisition, P.M.-A. and C.M.H. All authors have read and agreed to the
published version of the manuscript.
Funding: This research was funded by Escuela Politécnica Nacional, Quito, Ecuador [Project PIJ 17-04,
la ValSe-Food-CYTED] (Ciencia y Tecnología para el Desarrollo, Ref. 119RT0567) and Food4ImNut
(PID2019-107650RB-C21) from the MCIN/AEI/10.13039/501100011033, Spain).
Data Availability Statement: Data is contained within the article.
Conflicts of Interest: The authors declare no conflict of interest.

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