Scientia Agropecuaria 14(1): 49-57 (2023) Guillén Sánchez et al.

SCIENTIA
AGROPECUARIA Facultad de Ciencias
Scientia Agropecuaria Agropecuarias

Universidad Nacional de
Web page: http://revistas.unitru.edu.pe/index.php/scientiaagrop Trujillo

RESEARCH ARTICLE

Extraction of bioactive compounds from Peruvian purple corn cob by high

isostatic pressure
Jhoseline Stayce Guillén Sánchez1, 2 * ; Cinthia Baú Betim Cazarin3 ; Miriam Regina Canesin3 ;
Felix G. R. Reyes4 ; Amadeu Hoshi Iglesias5 ; Marcelo Cristianini1
1
Faculty of Food Engineering, Department of Food Technology, State University of Campinas, UNICAMP, P.O.Box. 6121, 13083-862,
Campinas, S.P. Brazil.
2
Formative Research and Teaching Research Program, Research Vice-rectorate, César Vallejo University, Av. Larco 1770, Trujillo, La
Libertad, Perú.
3
Faculty of Food Engineering, Food and Nutrition Department, University of Campinas, UNICAMP, 13083-862, Campinas, SP, Brazil.
4
Faculty of Food Engineering, Department of Food Science, University of Campinas, UNICAMP, Rua Monteiro Lobato, 80, 13083-862,
Campinas, SP, Brazil.
5
ApexScience Laboratory, Marechal Rondon Avenue, 2148, Jd. Chapadão, Campinas - SP, 13063-001.

* Corresponding author: [email protected] (J. S. Guillén Sánchez).

Received: 7 December 2022. Accepted: 5 February 2023. Published: 27 February 2023.

Abstract
The purple corn cob is an agro-industrial by-product that contains high-value bioactive compounds, which makes its use attractive for the
development of extraction processes. The aim of this research was to extract the bioactive compounds from the purple corn cob by high
isostatic pressure at different processing temperatures. Pressures of 0.01 MPa, 250 MPa, 450 MPa and 650 MPa for 3 minutes and at
temperatures of 25 °C, 45 °C and 65 °C were used. High pressure extraction was compared with conventional extraction (2.5 h at 25°C).
The purple corn cob extract obtained by isostatic processing at 650 MPa and 65 °C presented high antioxidant activity and content of
bioactive compounds, unlike the conventional extraction of 2.5 h and 65 °C (p < 0.05). However, it did not show a significant difference
with the extract obtained at 450 MPa and 45 °C (p > 0.05). Seven different anthocyanins were identified by liquid chromatography in the
extracts obtained by high isostatic pressure (650 MPa at 65 °C) and hydroalcoholic maceration (2.5 h at 65 °C), mainly cyanidin-3-glucoside,
pelargonidin-3-glucoside and peonidin-3-glucoside, and their respective malonyl derivatives. The high isostatic pressure increased the
extraction of bioactive compounds by more than 50% and obtained them in shorter times, thus appearing as a new alternative, and eco-
friendly method for the extraction of bioactive compounds from plant tissues.
Keywords: Purple corn cob; High pressure; Bioactive compound; Antioxidant activity; Anthocyanins.

DOI: https://doi.org/10.17268/sci.agropecu.2023.005

Cite this article:

Guillén Sánchez, J. S., Cazarin, C. B. B., Canesin, M. R., Reyes, F. G. R., Iglesias, A. H., & Cristianini, M. (2023). Extraction of bioactive
compounds from Peruvian purple corn cob by high isostatic pressure. Scientia Agropecuaria, 14(1), 49-57.

the pharmaceutical industry to control cardiovas-

1. Introduction cular diseases and/or diseases related to oxidative
Purple corn is an agricultural product native to the stress of the cells (Lao et al., 2017).
Peruvian Andes, cultivated mainly in the mountains The conventional techniques used to extract
and coast of Peru (National Institute of Agrarian bioactive compounds from purple corn are through
Innovation, 2007). The purple corn ear is made up heat treatment, at temperatures between 50 °C and
of 15% grain and 85% cob, which have purple 70 °C, and periods of time that vary between 30
pigmentation. Due to this pigmentation, purple minutes to 60 minutes (Rafael & Castro, 2023). The
corn presents considerable quantities of bioactive amount of bioactive compounds obtained by this
compounds that are found in greater proportions technique will depend mainly on the extraction time
in the cob than in the pericarp of the grain (Monroy and the solid:solvent ratio; considering that the
et al., 2016a). These bioactive compounds can be solvent with the highest extraction yield reported by
used in the food industry as food colorants and in the literature is acidified methanol. However,
-49-
Scientia Agropecuaria 14(1): 49-57 (2023) Guillén Sánchez et al.

methanol is a toxic solvent, so the food industry be stable at an isostatic pressure of 600 MPa.
prefers to use ethanol, relatively reducing the Researchers infer that vitamin K (phylloquinone) is
extraction yield. less susceptible to oxidative processes than the
Surco et al. (2022) compared the content of stronger antioxidants present in the juice, however,
phenolic compounds of Ricinus communis L there is a lack of literature on the effects of HIP on
extracts obtained by conventional techniques such vitamin K. The findings from this study also echo the
as ethanolic maceration and hydroalcoholic general consensus that HIP results in better imme-
maceration. The content of phenolic compounds in diate nutrient retention than thermal treatment.
the hydroalcoholic extracts was 20.98 ± 0.10 mg Therefore, the objective of this study was to extract
gallic acid/g sample, while in the ethanolic extracts the bioactive compounds from the purple corn cob
it was 102.30 ± 1.33 mg gallic acid/g sample. The by high isostatic pressure at different processing
latter also presented 96% more flavonoid content. temperatures, as alternatives to the aqueous,
Likewise, both extracts contain metals with antioxi- alcoholic and hydroalcoholic maceration technique,
dant activity such as copper, magnesium, zinc and which use large volumes of solvents, energy
calcium. In this sense, the researchers highlight its expenditure, because long times are required for
potential to carry out pre-clinical phase studies. extraction that can be from 8 hours to 8 days at
On the other hand, the high extraction tempera- room temperature (Arroyo et al., 2008).
tures have a negative effect on bioactive com-
pounds, since it increases the incidence of pigment 2. Materials and methods
loss, causing organoleptic and nutritional changes Conditioning of the sample
in the extracts (Villarroel et al., 2020; Radziejewska The purple corn was acquired from a popular
et al., 2020). To minimize the degradation of market located in the city of Chimbote, Ancash
bioactive compounds in plant materials, innovative region, Peru. The purple corn cob was dried in an
technologies have been investigated that are oven with air circulation at 50 °C for 24 hours, cut
respectful of the environment, such as high and ground to reduce the particle size, and sieved
pressure, microwaves and ultrasound, which reduce through a 60-mesh sieve for 5 minutes. The purple
the use of solvents and the impact of high corn cob powder was placed in low-density
temperatures (Terzića et al., 2023). polyethylene (LDPE) / ethylene vinyl alcohol (EVOH)
Of the eco-friendly technologies, the high-pressure bags and sealed using a vacuum sealer (Baseline P
technologies can achieve better extractions of the 100 / 300 w, MULTIVAC, Germany). The samples
bioactive compounds, since they do not affect the were stored at 4 °C until used.
nutritional quality, covalent bonds or thermolabile
Physical characteristics and composition of the
compounds (Barba et al., 2013). These compounds
purple corn cob powder
are not affected because the high pressures favor
The particle size was determined using a particle
the creation of new bonds in the system through
analyzer (SIROCCO 2000, MALVER INSTRUMENTS,
biochemical reactions, and, as a consequence, the
ENGLAND). The moisture and ash contents were
existing bonds are maintained with increasing
determined by gravimetric methods (AOAC, 2005);
pressure. Aghajanzadeh et al. (2022), mention that
the crude protein content was determined using the
high hydrostatic pressure technology (400 MPa -
Kjeldahl method (IAL, 2008) with a conversion factor
600 MPa / 15 min) improves the luminosity and
of 6.25; and the lipid content was determined using
chroma of blood orange juice by keeping the
the Bligh & Dyer (1959) method.
cyanidin-3-glucoside content constant. On the
other hand, isostatic pressures of 250 MPa at 65 °C Conventional extraction (CE)
for 3 minutes improve the content of total The corn cob extracts were prepared using a
carotenoids (36.84%), total anthocyanins (36.18%) powder: solvent ratio of 1:30 (w / v), and the solvent
and antioxidant capacity (34.57%), however, at used was a 20% (v / v) ethanol-water solution
higher pressures (≥ 600 MPa) the antioxidant acidified to pH 2 with 1 N HCl. The mixture was
activity can be reduced by approximately 30%. placed in flexible LDPE / EVOH / activated carbon
Koo et al. (2023) confirmed that high hydrostatic packages, sealed using a vacuum sealer and heated
pressure technology (600 MPa / 20 minutes) does in a thermostatic bath (MOD 116, FANEM ®, Brazil)
not significantly compromise the nutrient quality of for 2.5 hours at temperatures of 25 °C, 45 °C and
bok choy juice in terms of total antioxidant capacity, 65 °C. The extracts were filtered through Whatman
vitamin C, carotenoids, isothiocyanates (ITCs) and filter paper No. 1 under vacuum, placed in amber
vitamin K. In addition, ITCs and vitamin K, important bottles, covered with aluminum foil and stored at 4
nutrients in green leafy vegetables, were found to °C.

-50-
Scientia Agropecuaria 14(1): 49-57 (2023) Guillén Sánchez et al.

Extraction by high isostatic pressure (HIP) Singleton & Rossi (1965), with some modifications.
The HIP processes were carried out in a pilot scale 0.5 mL aliquots were taken and transferred to
high isostatic pressure equipment (MFF 2L-700, amber tubes, 2.5 mL of the Folin-Ciocalteu solution
Avure Technologies Inc, USA). The temperature of was added, and the volume was completed (10 mL)
the chamber was controlled by a K-type with the extraction solvent used in the extraction
thermocouple and the pressure by a pressure (20% ethanol-water / pH 2). The samples were left
transducer, using a computer for data acquisition. to rest in the dark for 5 minutes, 2 mL of a 7.5%
The corn cob powder: solvent mixtures were sodium carbonate solution was added, and the
prepared in a 1:30 (w / v) ratio, placed in flexible tubes were left in the dark for a further 2 h. The
packaging, sealed using a vacuum sealer and absorbance was measured at 760 nm using a
subjected to high isostatic pressure for 3 minutes, spectrophotometer (DU® 800 UV/Visible Spectro-
varying the pressures (250 MPa, 450 MPa and 650 photometer, Beckman Coulter, USA). The phenolic
MPa) and temperatures (25 °C, 45 °C and 65 °C). compound content was the concentration read
The extracts were filtered through Whatman filter multiplied by the corresponding dilution factor and
paper No. 1 under vacuum, placed in amber bottles, the results were expressed in milligrams of gallic
covered with aluminum foil and stored at 4 °C. acid (GAE) per gram of dry weight or mg GAE/g dry
weight.
Color Analysis
The color of the extracts was measured using a In vitro study of the antioxidant activity of purple
digital colorimeter (HUNTERLAB, UltraScan PRO, corn cob extracts processed by high isostatic
USA) based on the CIELAB color space system (L *, pressure and static maceration
a * and b *) (Montes et al., 2005). The cylindrical
coordinates Cab * (chroma) and hab (Hue angle) Oxygen Radical absorbance capacity - ORAC
were calculated from the parameters L *, a * and b The ORAC procedure was carried out according to
*, which define the intensity and tone of the samples Ou et al. (2013) in 75 mM phosphate buffer (pH 7.4),
according to equations 1 and 2, respectively. with a final reaction mixture of 200 uL. Twenty-five
microliters of the antioxidant solutions plus 150 uL
𝐶𝑎𝑏 ∗= [(𝑎∗ )2 + (𝑏∗ )2 ]0.5 (Eq.1) of a 11.12 x 10-2 M fluorescein solution were placed
𝐻 𝑎𝑏 = 𝑎𝑟𝑐𝑡𝑎𝑛 (𝑏∗ 𝑎∗ ) (Eq.2)
in the microplate wells, the microplate preincubated
Determination of the total monomeric anthocyanin for 10 min at 37 °C and twenty-five microliters of
content (TAC) AAPH (2, 2’-azobis (2-methylpropionamidine)
The TAC was determined using the AOAC dihydrochloride) then added rapidly to each well
differential pH method (2005). Aliquots of the using a multichannel pipet. The readings were
extract were diluted in amber tubes with 0.025 M carried out using a microplate reader (Synergy HT,
potassium chloride (pH 1) and 0.4 M sodium acetate Biotek® Instruments Inc., USA) and the
(pH 4.5) buffer solutions, and the absorbance of the fluorescence recorded every minute for 120 min,
solutions determined in the visible region at 520 nm the microplate being automatically shaken before
and 700 nm using a spectrophotometer (UV-VIS each reading. The results were expressed in μmol
DU® 800, Beckman Coulter, USA). The anthocyanin of Trolox (TE) per gram of dry weight.
content was expressed as cyanidin 3-O-glucoside
Ferric Reducing Antioxidant Power assay - FRAP
according to the following equation 3.
In this case, the antioxidant activity was determined
Where ‘ε’ is the molar absorptivity, ‘l’ is the length
according to Benzie & Strain (1996). The FRAP
of the passage through the cell, ‘A’ is the
reagent was prepared daily by mixing 2.5 mL
absorbance, ‘MW’ is the molecular weight of the
acetate buffer (0.3 M pH 3.6), 2.5 mL of a 10 mM
reference standard and ‘DF’ is the dilution factor.
TPTZ solution (2,4,6-Tris (2-pyridyl) -s-triazine) and
The equation was converted to express the results
2.5 mL of a 20 mM aqueous solution of ferric
in milligrams of cyanidin 3 glucoside per gram of
chloride, and maintained at 37 ° C. The test
dry weight or mg Cy3G/g dry weight.
solutions were prepared by mixing a 90 μL aliquot
𝐴𝑛𝑡ℎ𝑜𝑐𝑦𝑎𝑛𝑖𝑛 (𝑚𝑔 𝐶𝑦𝑎𝑛𝑖𝑑𝑖𝑛 3
𝐺𝑙𝑢𝑐𝑜𝑠𝑖𝑑𝑒
)= of each extract dilution, 270 μL distilled water and
2.7 mL of FRAP reagent, homogenized using a tube
𝐿
𝐴 𝑥 𝑀𝑊 𝑥 𝐹𝐷 𝑥 1000/ 𝜀 𝑥 𝑙 (Eq.3)
shaker and maintained in a water bath at 37°C. The
Determination of the total phenolic compound absorbance was measured at 595 nm after 30 min
content (TPC) using a microplate reader (Synergy HT, Biotek®
The TPC content was determined using the Folin- Instruments Inc., USA) and the results expressed in
Ciocalteu colorimetric method according to μmol of Trolox (TE) per gram of dry weight.

-51-
Scientia Agropecuaria 14(1): 49-57 (2023) Guillén Sánchez et al.

Identification of the anthocyanins in the purple corn Statistical analysis

cob extracts by Ultra-performance liquid The measurements obtained were evaluated using the
chromatography tandem mass spectrometry statistical program IBM SPSS v. 22 (SPSS Inc., Chicago,
(UPLC-MS/MS) USA), and analysis of variance (ANOVA) and Tukey's
An ultra-performance liquid chromatography test method were used to compare means. A level of
system (Waters Xevo I-Class, Milford, USA) coupled significance p < 0.05 was established.
to a tandem mass spectrometry detector (Waters
Xevo TQD, Milford, USA) with electrospray source 3. Results and discussion
ionization (ESI) in the negative mode was used to Characterization of the purple corn cob powder
identify the anthocyanins in the purple corn cob The protein, lipid and ash contents were similar to
extracts. The chromatographic separation was Cerro (2009) in purple corn cob with 11% and 9%
carried out using an ACQUITY UPLC® HSS T3 moisture contents, respectively. Another important
column (100 mm x 2.1 mm x 1.8 µm, Waters, Milford, fraction was the carbohydrate content, which was
USA) with a flow rate of 0.6 mL/min. The mobile 79.33%, since some pigments, such as simple
phase was composed of solvent A (0.1% TFA in anthocyanins, can be found esterified to one or several
water) and solvent B (acetonitrile). The gradient carbohydrates, or acylated anthocyanins, which can be
started at 10% B, rose to 35% over 5 min, and then found esterified to carbohydrates and an acyl radical
increased to 100% B in 7 min. The MassLynx (Strack & Wray, 1989).
software (Waters Co. Milford, USA) version 4.1 was
used for instrument control and data processing. Effect of HIP on the TAC of purple corn cob extracts
The samples were analyzed in the MS scan mode, Figure 1 shows the effects of HIP on the TAC values
and the compound masses were extracted (mg Cy3G/g dry weight) in the purple corn cob
according to Pascual-Teresa et al. (2002) using the extracts processed at three temperatures, as
following operating conditions: capillary voltage 3.5 compared to the contents determined using CE and in
kV, source temperature 150 °C, desolvation tempe- the non-pressurized extracts obtained at the same
rature 550 °C, cone gas flow 20 L/h and de- temperatures. Parameters such as the solvent and the
solvation gas flow 900 L/h. solid: solvent ratio were kept constant.

Figure 1. TAC of the extracts obtained by different processing methods and at different temperatures. CE for 2.5 hours, non-pressurized
at 0.01 MPa for 3 minutes and HIP (250 MPa, 450 MPa and 650 MPa) for 3 minutes. The bars are means ± standard deviation (n = 3).
Means with the same lower-case letters in the same column indicate that there is no significant difference (p > 0.05, Tukey test) between
the extracts subjected to the different processing methods, considering the same temperature. Different uppercase letters mean there is a
significant difference (p < 0.05, Tukey test) for different temperatures considering the same processing method.

-52-
Scientia Agropecuaria 14(1): 49-57 (2023) Guillén Sánchez et al.

The HIP processes at 650 MPa / 3 minutes at sample implies induced damage to the cell matrix,
temperatures of 25 °C and 45 °C presented 128% greater permeability and consequently the release
and 71% more TAC (p < 0.05), respectively than the of bioactive compounds bound to the matrix
processes at 250 MPa / 3 minutes at 25 °C and 45 (Siddiqui et al., 2023). On the other hand, Tatiane
°C, respectively, also presented 28% and 26% more et al. (2023) mention that the high pressure also
TAC, respectively, than the extracts obtained by CE allows the solvent to remain in a liquid state and
treated for 2.5 hours at 25 °C and 45 °C (p < 0.05), above its boiling point, being sufficiently capable of
respectively (Figure 1). The effect of HIP on the helping it penetrate the plant matrix and achieve
extraction of phytochemicals can be explained by greater extraction.
the rapid diffusion process that causes pressure, Increases in temperature also showed significant
facilitating disruption of the plant cells and their positive effects (p < 0.05) on the non-pressurized,
contact with the solvent used (García et al., 2016). HIP and CE processes. The increases in temperature
The results obtained in the present study for the increase the solubility of the solvent; according to
TAC in the extracts were higher than those reported Barba et al. (2017), such increases can be favorable
by Muangrat et al. (2018) for Thai purple corn cob since they have the selective ability to extract certain
extracts processed by ultrasound (ultrasonic thermolabile substances, but higher temperatures
probes, 20 kHz / 500 W / 30 min), and by can be aggressive, leading to alterations in the
Itthisoponkul et al. (2018) for aqueous extracts of substances or even their decomposition.
Thai purple corn cob processed by HIP (200 MPa /
25 °C / 15 minutes). According to Dukić et al. (2022),
Effects of HIP on the TPC of purple corn cob extracts
ultrasonic extraction through ultrasonic probes
implies direct contact of the probe with the analyte Figure 2 shows the effects of HIP on TPC (mg GAE/
and constant cooling of the sample, therefore, this g dry weight), in purple corn cob extracts processed
can lead to the degradation of the thermolabile at three temperatures, as compared to the contents
components and thus affect the reduction of the found in extracts obtained using CE and in non-
content of valuable molecules; while in high pressurized extracts obtained at the same
isostatic pressure, the pressure transfer being temperatures. Parameters such as the solvent and
simultaneous and uniform at all points of the solid: solvent ratios were maintained constant.

Figure 2. TPC of the extracts obtained by different processing methods at different temperatures. CE for 2.5 hours, non-pressurized at 0.01
MPa for 3 minutes and HIP (250 MPa, 450 MPa and 650 MPa) for 3 minutes. The bars are the means ± standard deviation (n = 3). Means
with the same lower-case letters in the same column indicate that there is no significant difference (p > 0.05, Tukey test) between the
extracts subjected to different processing methods, considering the same temperature. Different uppercase letters mean there is a
significant difference (p < 0.05, Tukey test) between the different temperatures, considering the same process.

-53-
Scientia Agropecuaria 14(1): 49-57 (2023) Guillén Sánchez et al.

Figure 2 shows that at a temperature of 25 °C and cylindrical chromatic space system, values between
65 °C, the increment in isostatic pressure from 250 180º and 270º indicate a trend from green to blue
MPa to 650 MPa, significantly (p < 0.05) increased (Padrón et al., 2012).
TPC contents by 51% and 12%, respectively. The
TPC content in the extracts obtained at 650 MPa/25 In vitro study of the antioxidant activities of the
°C/3 minutes was not statistically different from the purple corn cob extracts obtained by HIP and CE
TPC content of the extracts obtained by CE at 2.5 Table 1 shows a comparison of the antioxidant
hours/45 °C (p > 0.05). activities (AA) of the non-pressurized extracts and
The results obtained for the TPC in the extracts were those obtained by HIP and CE under conditions that
higher than those reported by Muangrat et al. showed high bioactive compound contents. The AA
(2018) and by Itthisoponkul et al. (2018). According values of the extracts were determined using the
to Dunford et al. (2010), the pressure-temperature ORAC and FRAP assays and the results were
interaction can act synergistically, since the pressure expressed in μM Trolox (TE) / g dry weight in both
facilitates contact between the matrix and solvent, cases.
producing the extracts quicker than the
conventional extraction, while an increase in The AA values of the extracts were determined
temperature decreases the dielectric constant, using the ORAC and FRAP assays and the results
equalizing the polarity of the solvent and the were expressed in μM Trolox (TE) / g dry weight in
compounds, hence extracting them more easily. In both cases. Of the extracts obtained by CE, those
the results obtained for the TAC and TPC contents obtained in 2.5 hours/ 65 °C showed the highest AA
in extracts obtained by high pressure technology, it values according to ORAC. The extract obtained by
can be seen that the HIP process at 450 MPa / 45 HIP at 650 MPa / 65 °C / 3 minutes, showed high
°C was efficient in increasing the extraction of TAC values for AA, 30% and 55% more than the extract
and TPC. obtained by CE in 2.5 hours/ 65 °C (p < 0.05),
according to FRAP and ORAC, respectively.
Effects of HIP on the color of the purple corn cob The present results suggest that using high
extracts pressures can reduce the times from hours to
For the temperatures of 25 °C, 45 °C and 65 °C, minutes and achieve extracts with high AA values.
increases in HIP from 250 MPa to 650 MPa did not According to Tian et al. (2018) polar antioxidant
significantly alter (p > 0.05) the values of the compounds have a greater reducing capacity,
coordinates a * and b*, chroma (C*) or brightness inferring that the AA of the extracts obtained by
(L*), obtaining extracts with tendencies between high-pressure technologies is due to the greater
blue and red. These values were different from presence of phenolic compounds. The
those reported by Monroy et al. (2016b) for methodologies used revealed that the AA values of
Peruvian purple corn cob extracts obtained using the purple corn cob extracts obtained by high
supercritical fluids (400 bar / 50 °C / 390 minutes / isostatic pressure technology were significantly
three sequential extractions with SCO2, ethanol and higher than the values in the extracts obtained by
water). For all these processes, the values obtained CE, results which are consistent considering that the
for h° in the extracts were in the range from 247.08° extracts obtained using high-pressure presented
to 274.02°, and according to the CIE-L* C* hº higher TPC contents (Saikaew et al., 2018).

Table 1
Comparative of the antioxidant activity as determined by the ORAC and FRAP assays in the conditions of greater efficiency of the
technologies

Antioxidant Activity (μM TE/g dry weight)

Process Variables FRAP ORAC
0.01 MPa/25 °C/3 minutes 312.51±11.14 e 980.55±42.32 de
Non-pressurized 0.01 MPa/45 °C/3 minutes 480.87±41.67 de 1222.77±74.50 bcd
0.01 MPa/65 °C/3 minutes 753.34±16.03 abc 1145.39±46.63 cd
650 MPa/25 °C/3 minutes 722.48±64.32 abc 860.82±12.86 ef
HIP 650 MPa/45 °C/3 minutes 788.60±25.64 abc 1225.45±61.94 bcd
650 MPa/65 °C/3 minutes 880.66±79.44 ab 1056.43±43.35 cde
2.5 hours/25 °C 591.96±39.57 cd 849.31±22.36 ef
CE 2.5 hours/45 °C 509.09±44.12 de 999.52±16.82 ed
2.5 hours/65 °C 676.75±9.95 bcd 682.29±21.64 f
* The data are presented as the means ± standard deviation (n = 3). Same letters in the column indicate that there are no significant differences among the
different process conditions (p < 0.05, Tukey test).

-54-
Scientia Agropecuaria 14(1): 49-57 (2023) Guillén Sánchez et al.

Table 2
Areas of the chromatographic bands obtained by technology for the compounds found in the extracts of purple corn

Peak Compounds CE HIP

1 Unknown 120026 172428
2 Cyanidin-3-glucoside 11242858 6593593
3 Pelargonidin-3-glucoside 2497220 1573643
4 Peonidin-3-glucoside 6995596 4349066
5 Cyanidin-3-(6-malonylglucoside) 7855386 3709423
6 Pelargonidin-3-(6-malonylglucoside) 3792038 2408920
7 Peonidin-3-(6-malonylglucoside) 4622761 2850615
* CE process at 2.5 hours/65 °C and HIP process at 650 MPa/65 °C/3 minutes.

Figure 3. Mass chromatograms obtained by UPLC-MS/MS, highlighting the majoritarian anthocyanins in purple corn cob extract using HIP
process (650 MPa/65 °C/3 minutes).

UPLC–MS/MS analysis of the anthocyanins in the degradation than their non-acylated forms and,
purple corn cob extracts obtained by HIP and CE consequently, the extracts maintained greater color
UPLC-MS/MS was applied to the study of the stability. According to Falcão et al. (2003), acylated
anthocyanin compositions of the purple corn cob anthocyanins, which have cinnamic and/or malonic
extracts obtained by high isostatic pressure and CE. acid residues esterified to sugars, and non-acylated
The compounds were identified by comparing their anthocyanins can be found in samples of plant
retention times with those reported by Pascual- origin. Thus, the anthocyanin composition of the
Teresa et al. (2002). Peak 1 may reveal the presence corn cob, as well as that of the extracts obtained,
of a dimer formed by condensation between makes them a new alternative for use as natural
different flavonoids (Pascual-Teresa et al., 2002). In coloring compounds and for the dyeing of foods.
this study, the chromatographic profiles were
similar to those found by Pascual-Teresa et al. 4. Conclusions
(2002) in a commercial extract from purple corn The study demonstrated that the application of
(Figure 3). isostatic pressures greater than 450 MPa for 3
The areas of the chromatographic bands of the an- minutes allows obtaining purple corn cob extracts
thocyanins obtained using the different extraction with a high content of bioactive compounds and
processes (CE and HIP) are shown in Table 2. high antioxidant activity of oxygen radical
The presence of acyl groups in the anthocyanin absorption capacity. The controlled synergistic
molecules makes them more stable to thermal effect of the isostatic pressure and temperature

-55-
Scientia Agropecuaria 14(1): 49-57 (2023) Guillén Sánchez et al.

binomial increased the content of anthocyanins and Dukić, J., Hunić, M., Nutrizio, M., Režek, A. (2022). Influence of
High-Power Ultrasound on Yield of Proteins and Specialized
the content of phenolic compounds by 70% and Plant Metabolites from Sugar Beet Leaves (Beta vulgaris
60%, respectively, considering a temperature range subsp. vulgaris var. altissima). Applied Sciences, 12(18), 8949.
of 25 °C and 45 °C, however, it significantly affects Falcão, L., Barros, D., Gauche, C., & Bordignon, M. (2003).
Copigmentação intra e intermolecular de antocianinas: uma
the brightness and hue angle of the samples. Based
revisão. Boletim do Centro de Pesquisa de Processamento de
on the results, the combination of high isostatic Alimentos, Curitiba, 21(2), 351-366.
pressure with mild thermal methods can be García, J., González-Cebrino, F., Delgado, J., Cava, R., & Ramírez,
suggested to guarantee the highest efficiency of the R. (2016). High pressure assisted thermal processing of
pumpkin purée: Effect on microbial counts, color, bioactive
extraction process of bioactive compounds in plant compounds and polyphenoloxidase enzyme. Journal of Food
materials, which can be used as colorants or and Bioproducts Processing, 98, 124-132.
antioxidants in the development of functional IAL - Instituto Adolfo Lutz (2008). Procedimentos e determinações
gerais. In: Zenebon O, Pascuet N, Tiglea, P (eds.), Métodos
foods.
Físico-Químicos para análise de alimentos, pp.83-158.
Itthisoponkul, T., Naknan, P., Prompun, S., & Kasemwong, K. (2018).
Acknowledgements Evaluation of high pressure treatment for improvement of
This work was supported by the National Council for physicochemical and functional qualities in purple corncobs.
Scientific and Technological Development (CNPq) [grant International Food Research Journal, 25(1), 246-253.
numbers 132314/2017-7]. Koo, A., Xiang Chew, D., Ghate, V., Zhou, W. (2023). Residual
polyphenol oxidase and peroxidase activity in high pressure
ORCID processed bok choy (Brassica rapa subsp. chinensis) juice did
not accelerate nutrient degradation during storage.
J. S. Guillén Sánchez https://orcid.org/0000-0002-0899-6725
Innovative Food Science & Emerging Technologies, 103284.
C. B. B. Cazarin https://orcid.org/0000-0002-9849-2546
Lao, F., Sigurdson, G., & Giusti, M. (2017). Health Benefits of Purple
M. R. Canesin https://orcid.org/0000-0002-2892-3814
Corn (Zea mays L.). Phenolic Compounds. Comprehensive
F. G. R. Reyes https://orcid.org/0000-0003-0126-3817
Reviews in Food Science and Food Safety, 16, 234-246.
A. H. Iglesias https://orcid.org/0000-0003-1119-5627
Monroy, Y., Rodrigues, R., Sartoratto, A., & Cabral, F. (2016a).
M. Cristianini https://orcid.org/0000-0001-7740-0449
Influence of ethanol, water, and their mixtures as co-solvents
of the supercritical carbon dioxide in the extraction of
References phenolics from purple corn cob (Zea Mays L.). The Journal of
Aghajanzadeh, S., Ziaiifar, A., Verkerk, R. (2022). Effect of thermal Supercritical Fluids, 118, 11–18.
and non-thermal treatments on the color of citrus juice: A Monroy, Y., Rodriguez, R., Sartoratto, A., & Cabral, F. (2016b).
review. Food Reviews International, 3(1), 1-23. Extraction of bioactive compounds from cob and pericarp of
AOAC. Horwitz, W. (Ed). (2005). Official methods of Analysis of purple corn (Zea mays L.) by sequential extraction in fixed bed
AOAC International, 18th edition, AOAC International, extractor using supercritical CO2, ethanol, and water as
Maryland, USA. solvents. The Journal of supercritical fluids, 118, 10-19.
Arroyo, J., Raez, E., Rodriguez, M., Chumpitaz, V., Burga, J., De La Montes, C., Vicario, I., Raymundo, M., Fett, R., & Heredia, F. (2005).
Cruz, W., & Valencia, J. (2008). Antihypertensive and Application of tristimulus colourimetry to optimize the
antioxidant activity of the atomized hydroalcoholic extract of extraction of anthocyanin from Jaboticaba (Myricia Jaboticaba
purple corn (Zea Mays L.) in rats. Revista Peruana de Medicina Berg.). Food Research International, 38, 983–988.
Experimental y Salud Pública, 25(2), 195-199. Muangrat, R., Pongsirikul, I., & Blanca, P. (2018). Ultrasound
Barba, F., Esteve, M., & Frigola, A. (2013). Physicochemical and assisted extraction of Anthocyanins and total phenolic
nutritional characteristics of blueberry juice after high compounds from dried cob of purple waxy corn using
pressure processing. Food Research International, 50, 545- response surface methodology. Journal of food processing
549. and preservation, 42(2), e13447.
Barba, F, Putnik, P., Bursac, D., Poojary, M., Roohinejad, S., National Institute of Agrarian Innovation – INIA (2007). Boletín
Lorenzo, J., & Koubaa, M. (2017). Impact of conventional and informativo Maíz INIA 615 Negro Canaán.
non-conventional processing on prickly pear (Opuntia spp.) https://repositorio.inia.gob.pe/bitstream/20.500.12955/648/1
and their derived products: From preservation of beverages /Trip-Maiz-INIA613.pdf
to valorization of by-products. Trends in Food Science & Ou, B., Chang, T., Huang, D., & Prior, R. (2013). Determination of
Technology, 67, 260-270. total antioxidante capacity by oxigen radical absorvace
Benzie, F., & Strain, J. (1996). The Ferric Reducing Ability of Plasma capacity (ORAC) using fluorescein as the fluorescence probe:
(FRAP) as a Measure of ‘‘Antioxidant Power’’: The FRAP Assay. First action 2012.23. Food composition and additives, 96(6),
Analytical Biochemistry, 239, 70-76. 1372-1376.
Bligh, E., & Dyer, W. (1959). A rapid method of total lipid extraction Padrón, C., Padrón, G., Montes, A., & Oropeza, R. (2012).
and purification. Canadian Journal of Biochemistry and Determinación del color en epicarpio de tomates
Physiology, 37(8), 911-917. (Lycopersicum esculentum Mill.) con sistema de visión
Rafael, E., & Castro, D. (2023). Extracción y cuantificación de computarizada durante la maduración. Agronomía
Antocianinas de maíz morado (Zea Mays L.) utilizando dos Costarricense, 36(1), 97-111.
solventes a diferentes temperaturas y tiempos de extracción. Pascual-Teresa, S., Santos-Buelga, C., & Rivas-Gonzalo, J. (2002).
Revista Latinoamericana de Difusión Científica, 5(8), 47-80. LC-MS Analysis of anthocyanins from purple corn cob.
Cerro, R. (2009). Extração e caracterização do pigmento do Journal Science Food Agriculture, 82, 1003-1006.
sabugo de milho roxo (Zea Mays L.) e sua aplicação em Radziejewska, E., Szwengiel, A., Ratajkiewicz, H., & Nowak, K.
alimentos (Dissertação de Mestrado). Universidade Estadual (2020). Effect of ultrasound, heating and enzymatic pre-
de Campinas, Campinas, Brasil. treatment on bioactive compounds in juice from Berberis
Dunford, N., Imak, S., Jonnala, R. (2010). Pressurized solvent amurensis Rupr. Ultrasonics – Sonochemistry, 63, 104-971.
extraction of policosanol from wheat straw, germ and bran. Saikaew, K., Lertrat, K., Meenune, M., & Tangwongchai, R. (2018).
Food Chemistry, 119(3), 1246-1249. Effect of high-pressure processing on colour, phytochemical

-56-
Scientia Agropecuaria 14(1): 49-57 (2023) Guillén Sánchez et al.

contents and antioxidant activities of purple waxy corn (Zea process variables and extract characterization. The Journal of
mays L. var. ceratina) kernels. Food Chemistry, 243, 328–337. Supercritical Fluids, 193, 105813.
Siddiqui, S., Schulte, H., Pleissner, D., Schönfelder, S., Kvangarsnes, Terzića, M., Majkićb, T., Zengin, G., Beara, I., Céspedes-Acuña, C.,
K., et al. (2023). Transformation of Seafood Side-Streams and Čavić, D., & Radojkovica, M. (2023). Could elderberry fruits
Residuals into Valuable Products. Foods, 12, 422. processed by modern and conventional drying and extraction
Singleton, V., & Rossi, J. (1965). Colorimetry of Total Phenolics with technology be considered a valuable source of health-
Phosphomolybdic-Phosphotungstic Acid Reagents. American promoting compounds? Food chemistry, 405, 134766.
Journal of Enology and Viticulture, 16(3), 144-158. Tian, Y., Puganen, A., Alakomi, H. L., Uusitupa, A., & Saarela, M.
Strack, D., & Wray, V. (1989) Anthocyanins. In: Harbone H (ed). (2018). Antioxidative and antibacterial activities of aqueous
Methods in Plant Biochemistry, Plant Phenolics. Academic ethanol extracts of berries, leaves, and branches of berry
Press, New York, pp. 325-356. plants. Food research international, 106, 291-303.
Surco, F., García, J., Valle, M., Panay, J., Bonifaz, M., et al. (2023). Villarroel, J., Sanabria, N., & Pérez, L. (2020). Bioactive compounds
Bioactive compounds and in vitro antioxidant activity of the and kinetic degradation of anthocyanins in extracts of
ethanolic and hydroalcoholic extract of Ricinus communis L. Hibiscus sabdariffa L. Revista Ciencia Tecnologia. Agrollanía,
(Higuerilla). Revista Cubana de Farmacia, 55(4), e840. 19, 18-24.
Tatiane, D., Da Silva, C., & Cláudio Da Costa, S. (2023). Pressurized
liquid extraction of compounds from Stevia leaf: Evaluation of

-57-