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Food Chemistry
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a r t i c l e
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Article history:
Received 16 October 2014
Received in revised form 6 March 2015
Accepted 10 March 2015
Available online 17 March 2015
Keywords:
Quinoa (Chenopodium quinoa Willd.)
Phenolic compounds
Betalains
Total antioxidant capacity
QUENCHER procedure
Peruvian Altiplano
a b s t r a c t
Physical features, bioactive compounds and total antioxidant capacity (TAC) of coloured quinoa varieties
(Chenopodium quinoa Willd.) from Peruvian Altiplano were studied. Quinoa seeds did not show a pure red
colour, but a mixture which corresponded to different fractal colour values (51.071.8), and they varied
from small to large size. Regarding bioactive compounds, total phenolic (1.233.24 mg gallic acid equivalents/g) and avonol contents (0.472.55 mg quercetin equivalents/g) were highly correlated (r = 0.910).
Betalains content (0.156.10 mg/100 g) was correlated with L colour parameter (r = 0.569), total phenolics (r = 0.703) and avonols content (r = 0.718). Ratio of betaxanthins to betacyanins (0.01.41) was
negatively correlated with L value (r = 0.744). Whereas, high TAC values (119.8335.9 mmol Trolox
equivalents/kg) were negatively correlated with L value (r = 0.779), but positively with betalains
(r = 0.730), as well as with free (r = 0.639), bound (r = 0.558) and total phenolic compounds (r = 0.676).
Unexploited coloured quinoa seeds are proposed as a valuable natural source of phenolics and betalains
with high antioxidant capacity.
2015 Elsevier Ltd. All rights reserved.
1. Introduction
Quinoa (Chenopodium quinoa Willd.) plants from the Andean
region in South America possess a huge genetic variability which
allows its adaptation and growth under adverse environmental
conditions, such as drought, hail, frost and high altitude (Aguilar
& Jacobsen, 2003; Jacobsen, 2003). Quinoa seed has been recognized as an extremely nutritious grain all over the world, due to
both its relatively high amount (compared to cereals) and the ne
quality of its proteins, as regards essential amino acids content;
quinoa seed also contains essential fatty acids and minerals
(Vega-Galvez, Miranda, Vergara, Uribe, Puente, & Martinez,
2010). Besides, its high total antioxidant capacity (Hirose, Fujita,
Ishii, & Ueno, 2010; Repo-Carrasco-Valencia, Hellstrm, Pihlava,
& Mattila, 2010; Tang et al., 2015) is related to its high phenolic
content which can vary depending on genetic variability and
environmental conditions (Repo-Carrasco-Valencia et al., 2010).
Corresponding author.
E-mail address: lcondezotm@hotmail.com (L. Condezo-Hoyos).
http://dx.doi.org/10.1016/j.foodchem.2015.03.029
0308-8146/ 2015 Elsevier Ltd. All rights reserved.
84
85
Fig. 1. Colour parameters of coloured quinoa seeds from Peruvian Altiplano on HSL space colour. (A) Lightness (%) and (B) Saturation (%) and Hue (sexagesimal grade) were
calculated from RGB values obtained from 25 images by histogram color analysis. Common names of the samples are: M1 = Cuchiwilla morado, M2 = Quinua rosada,
M3 = Quinua chica, M4 = Pasankalla dorado, M5 = Quinua real rosado, M6 = Pasankalla rojo, M7 = Quinua rosado, M8 = Cuchiwilla roja, M9 = Ayrampo, M10 = Panela,
M11 = Witulla roja, M12 = Cuchiwilla puaquinua and M13 = Vilacuyo. (For interpretation of the references to colour in this gure legend, the reader is referred to the web
version of this article.)
86
Fig. 2. PCA score plot biplot for the discrimination of the coloured quinoa based on
morphological and colour features. Score plot (samples) and loading (variable
measured) are superimposed. (For interpretation of the references to colour in this
gure legend, the reader is referred to the web version of this article.)
Table 1
Morphological and fractal colour features of coloured quinoa seeds (Chenopodium quinoa Willd.) from Peruvian Altiplano.
Sample
M1
M2
M3
M4
M5
M6
M7
M8
M9
M10
M11
M12
M13
Morphological features
Area (mm2)
Perimeter (mm)
Circularity
AR
2.52 0.32
2.94 0.44
2.85 0.34
4.71 0.59
3.55 0.40
3.53 0.36
3.72 0.54
2.31 0.20
2.41 0.25
4.21 0.47
2.46 0.32
3.15 0.37
3.16 0.40
5.95 0.40
6.50 0.49
6.33 0.39
8.17 0.52
7.15 0.44
7.07 0.39
7.61 0.91
5.69 0.27
5.83 0.30
7.80 0.45
5.90 0.41
6.70 0.40
6.75 0.43
1.89 0.13
2.03 0.16
1.97 0.11
2.52 0.17
2.19 0.14
2.22 0.13
2.24 0.17
1.78 0.08
1.81 0.10
2.45 0.13
1.87 0.13
2.16 0.13
2.16 0.13
1.69 0.11
1.83 0.13
1.83 0.13
2.37 0.15
2.06 0.12
2.02 0.10
2.10 0.15
1.65 0.07
1.69 0.09
2.18 0.14
1.66 0.11
1.85 0.13
1.86 0.15
0.89 0.01
0.87 0.01
0.89 0.01
0.88 0.01
0.87 0.02
0.89 0.01
0.82 0.12
0.90 0.01
0.89 0.01
0.87 0.03
0.88 0.01
0.88 0.01
0.87 0.01
1.12 0.05
1.11 0.04
1.08 0.04
1.06 0.04
1.06 0.04
1.10 0.04
1.07 0.03
1.08 0.03
1.07 0.04
1.12 0.05
1.13 0.05
1.17 0.06
1.16 0.09
56.9 1.5
68.9 3.3
59.7 1.5
64.9 4.4
51.0 0.5
65.9 2.4
70.8 4.2
60.6 4.0
60.4 4.1
62.9 5.2
57.8 6.7
61.3 3.3
71.8 7.8
87
III, IV) with decreasing levels of both, L value and fractal colour,
respectively (Fig. 2). To the best of our knowledge, this is the rst
time that the above physical parameters have been successfully
applied to group coloured quinoa samples.
3.2. Free, bound and total phenolics and avonols in coloured quinoa
seeds
Coloured quinoa seeds from Peruvian Altiplano showed free
phenolic values comprised between 1.23 and 3.41 mg GAE/g sample (Table 2), which are comparable to those found for red quinoa
samples (4.2 mg GAE/g sample) grown in Ontario (Canada) (Tang
et al., 2015), canihua (2.5 mg GAE/g sample dry weight) another
Altiplano pseudo-cereal that belongs to the same genus as the
quinoa seed (Abderrahim et al., 2012) and different types of
millet (1.46.3 mg ferulic acid equivalent/g defatted sample)
(Chandrasekara & Shahidi, 2010). Regarding bound phenolics content, coloured quinoa seeds from Peruvian Altiplano showed values
ranging from 1.28 to 4.52 mg GAE/g (Table 2) and a ratio of free to
bound phenolics between 0.5 and 2.0 (calculated from data in
Table 2). The former values were similar to those found (0.61.6)
for other previously studied Altiplanos coloured quinoa seeds
(Pasankalla, Roja de Coporaque and Witulla) (Repo-CarrascoValencia et al., 2010). On the contrary, bound phenolics in coloured
quinoa were lower than those found for canihua seed (8.8 mg/g
sample dry weight) (Abderrahim et al., 2012) and whole millet
types (0.415.9 mg ferulic acid/g defatted sample) (Chandrasekara
& Shahidi, 2010). However, the hydrolysis conditions (2 M HCl at
85 C for 1 h) employed to release bound phenolics, could also
increase furan derivatives that at very high concentration
(>3000 mg/kg) could react with FolinCiocalteau reagent leading
to the overestimation of bound phenolic content. In the present
study an efcient hydrolysis with methanol and sulphuric acid
(instead of hydrochloric acid), avoided furan derivatives formation
and released bound phenolics by more than vefold, compared to a
previous report using alkaline hydrolysis for nonextractable
polyphenols in cereal samples (Arranz & Saura Calixto, 2010). In
fact, a positive correlation (r = 0.838; p < 0.0001) between bound
phenolics and bound avonols, conrmed the lack of interferences
for furan derivatives, as detected with 2-aminoethyldiphenyl
borate colorimetric assay.
Previous studies have described that quinoa seeds are an exceptionally rich source of avonols, such as quercetin and kaempferol
(Repo-Carrasco-Valencia et al., 2010; Tang et al., 2015). In the
present study, we have also demonstrated a very high content of
total avonols (0.472.55 mg QE/mg sample) in quinoa coloured
seeds from Peruvian Altiplano (Table 2). The above avonols content was comparable to that of total avonoids for quinoa seeds
(white, red and black) commercialized in the Canadian market
(1.55.0 mg catechin equivalent/g quinoa sample) (Tang et al.,
2015). However, avonols content found in coloured quinoa seeds
from Peruvian Altiplano was markedly higher than total avonoids
(36.2 to 144.3 mg/100 g quinoa sample) of some other coloured
quinoa seeds (Pasankalla, Roja de Coporaque and Witulla) (RepoCarrasco-Valencia et al., 2010). Differences in extraction, hydrolysis procedures and analytical techniques are likely to explain the
content of total avonols found in the present study, and total
avonoids reported by Repo-Carrasco-Valencia et al. (2010).
Similarly, total avonols of coloured quinoa seeds from Peruvian
Altiplano (1937% of total phenolics, as calculated from Table 2)
were similar to the free avonoids in white, red and black genotypes of quinoa seeds commercialized in the Canadian markets,
which reached only approximately 30% of total phenolics (Tang
et al., 2015). In contrast, a very high avonoids percentage regarding total phenolics was reported for some red quinoa seeds from
Peruvian Altiplano: roja de coporaque (69.3%), witulla (69.5%) and
03-21-0093 (54.6%) (Repo-Carrasco-Valencia et al., 2010). These
differences could be explained by the short time (30 min) and
low acid concentration (approximately 1 M HCl) employed by
Repo-Carrasco-Valencia et al. (2010), which did not allow the full
extraction of bound phenolics from quinoa samples, predominantly composed by phenolic acids in cereals (Acosta-Estrada
et al., 2014).
A very high positive association was found between total phenolics and total avonols (r = 0.91, p < 0.0001). Another interesting
negative correlation was found between phenolic compounds and
L values (value r = 0.619, p = 0.024), which reected well that
both phenolics and pigments from coloured quinoa samples
are simultaneously extracted in methanol, and that an improvement
in phenolic compounds was accompanied by an increase in pigments content. Finally, a signicant and positive association between
free avonols and free phenolics (r = 0.677, p = 0.011), bound avonols and bound phenolics (r = 0.838, p < 0.0001), and total avonols
and total phenolics (r = 0.910, p < 0.0001) was found.
3.3. Betalains content of coloured quinoa seeds
The presence of betalains in red quinoa seeds remains controversial, despite quinoa belong to the same family as amaranth,
which seed contains low levels of amaranthine type of betacyanins
(1.4 mg/100 g). Repo-Carrasco-Valencia et al. (2010) do not detect
the presence of betacyanins in some Peruvian Altiplanos red
Table 2
Free and bound phenolics, avonols and total antioxidant capacity of coloured quinoa seeds (Chenopodium quinoa Willd.) from Peruvian Altiplano.
Sample
M1
M2
M3
M4
M5
M6
M7
M8
M9
M10
M11
M12
M13
Betalains (mg/100 g)
Free
Bound
Total
Free
Bound
Total
Bcy
Bx
Total
2.87 0.17
1.90 0.16
3.24 0.03
2.72 0.03
1.23 0.18
2.77 0.01
2.67 0.13
2.59 0.13
3.41 0.06
1.38 0.02
2.54 0.02
2.98 0.03
2.57 0.15
4.01 0.11
3.81 0.33
2.45 0.05
1.45 0.08
1.28 0.04
4.29 0.17
1.79 0.02
2.53 0.03
4.52 0.06
1.89 0.03
3.04 0.17
3.23 0.06
4.09 0.18
6.89 0.06
5.71 0.17
5.69 0.02
4.17 0.10
2.50 0.22
7.07 0.18
4.46 0.15
5.12 0.02
7.92 0.12
3.27 0.05
5.58 0.19
6.20 0.09
6.66 0.33
0.75 0.02
0.90 0.16
0.93 0.06
0.92 0.04
0.30 0.02
0.81 0.16
1.13 0.23
1.12 0.00
1.08 0.12
0.62 0.06
0.84 0.05
1.09 0.10
0.52 0.03
1.32 0.04
0.33 0.11
0.75 0.02
0.38 0.03
0.16 0.03
1.34 0.03
0.28 0.03
0.75 0.17
1.47 0.10
0.52 0.08
0.89 0.01
1.10 0.06
1.29 0.03
2.06 0.02
1.23 0.05
1.68 0.04
1.30 0.01
0.47 0.01
2.15 0.13
1.40 0.19
1.87 0.17
2.55 0.02
1.14 0.02
1.72 0.04
2.19 0.04
1.81 0.00
4.18 0.21
1.20 0.08
3.59 0.18
1.99 0.07
0.15 0.01
2.94 0.10
0.43 0.02
1.78 0.05
5.23 0.23
0.38 0.02
0.68 0.03
1.87 0.05
0.18 0.00
0.44 0.01
0.18 0.01
0.59 0.02
0.52 0.03
0.00 0.00
1.63 0.01
0.00 0.00
0.30 0.01
0.87 0.04
0.05 0.01
0.46 0.01
0.42 0.02
0.00 0.00
4.63 0.03
1.38 0.05
4.17 0.06
2.51 0.05
0.15 0.01
4.57 0.07
0.43 0.02
2.08 0.04
6.10 0.12
0.43 0.03
1.14 0.04
2.29 0.04
0.18 0.00
321.4 3.2
219.5 10.5
264.4 11.0
201.4 10.6
119.8 2.8
335.9 12.2
182.3 8.2
311.7 2.3
259.2 9.8
167.2 9.9
262.2 12.5
267.6 3.7
162.6 4.9
88
Fig. 3. Correlation between the total antioxidant capacity measured by QUENCHER-CUPRAC procedure and L value as colour parameter, total betalains and free, bound and
total phenolics and avonols in coloured quinoa seed from Peruvian Altiplano. A parametric Pearson procedure was used and p < 0.05 denote a statistical signicance. (For
interpretation of the references to colour in this gure legend, the reader is referred to the web version of this article.)
Fig. 4. PCA score plot biplot for the discrimination of the coloured quinoa based on
phenolics, betalains and total antioxidant capacity. Score plot (samples) and loading
(variable measured) are superimposed. The closer the samples are to a particular
variable the higher the positive correlation. (For interpretation of the references to
colour in this gure legend, the reader is referred to the web version of this article.)
89
4. Conclusion
3.4. Total antioxidant capacity of coloured quinoa seeds
In cereal and pseudo-cereal samples, a high phenolics amount is
covalently bound to the insoluble fraction, so that they cannot be
totally extracted by solvents (Abderrahim et al., 2012; AcostaEstrada et al., 2014; Arranz, Silvan, & Saura-Calixto, 2010).
Consequently, the antioxidant capacity of cereal and pseudo-cereal
samples, as measured by based-extracts traditional procedures
might be underestimated (Serpen et al., 2008). For this reason, a
direct procedure which does not require a previous extraction step
of antioxidants and based on copper (II) oxidizing reagents (Tufan
et al., 2013) has been used to assess the antioxidant capacity of
Peruvian Altiplanos coloured quinoa samples. In agreement with
their higher phenolic and betalain content, a very high total
antioxidant capacity (TAC), comprised between 119.8 2.8 and
335.9 12.2 mmol Trolox equivalent/kg was found for coloured
quinoa samples (Table 2). TAC of quinoa samples was markedly
higher than values reported for cereals as barley (26.35 0.36),
rye (16.21 0.25), wheat (13.44 0.46) and oat (10.46 0.23),
which were also measured using a CUPRAC chromogenic reagent
QUENCHER-CUPRAC (Tufan et al., 2013). In addition, TAC for
Peruvian Altiplanos coloured quinoa seeds was even higher than
that measured for canihua seed (71.0 1.7 mmol Trolox equivalent/kg sample dry weight). Nevertheless, this value was assessed
by using ABTS method (Abderrahim et al., 2012), which can provide lower TAC values than CUPRAC chromogenic reagent, due to
steric hindrance that reduces the kinetic of the reaction between
This study demonstrated for the rst time that coloured quinoa
seeds from Peruvian Altiplano are a rich source of free and bound
phenolic compounds and betalains. Moreover, these quinoa samples showed a very high antioxidant capacity compared to cereals
and as measured by QUENCHER-CUPRAC direct procedure. Overall,
these results strongly suggest that coloured quinoa seeds grown
under extreme conditions in the Peruvian Altiplano region might
be interesting as a natural source of functional food ingredients.
Acknowledgements
The authors thank Dr. Oldrich Zmeskal, Tomas Bzatek and
Martin Nezadal from Brno University of Technology in Brno
(Czech Republic) for supplying HarFA 5.5.30 software and Dr.
Pilar Ruprez from Instituto de Ciencia y Tecnologa de Alimentos
y Nutricin (ICTAN), Consejo Superior de Investigaciones
Cientcas (CSIC), Madrid, Spain for her critical review of the
manuscript.
90
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