Manuscrito Publicado 022017
Manuscrito Publicado 022017
Manuscrito Publicado 022017
Mango is one of the tropical fruits of greater production and consumption in the world, and a rich
source of bioactive compounds, with various functional properties such as antioxidant activity. In
Colombia, mango’s market is very broad and diverse. However, there are very few studies that
determined the content of bioactive secondary metabolites. The objective of this study was to evaluated
the content of different metabolites like Mangiferin, carotenoids, tannins, and the antioxidant capacity
by oxygen radical absorbance capacity (ORAC) methodology of six cultivars from the Colombian
Caribbean region, with total carotenoid values ranging from 24.67 to 196.15 mg of β-carotene/100 g dry
pulp; 84.30 to 161.49 mg Catequine eq./100 g dry pulp for the content of condensed tannins, and 91.80
to 259.23 mg/100 g dry pulp for mangiferin content. The ORAC methodology showed important
antioxidant activity results, such as the Chancleta variety with the highest value (2163.78 μmol
Trolox/100 g dry pulp). In conclusion, the evaluated mango varieties had promising results as functional
food of high nutraceutical value, being Chancleta, Criollo and Jobo varieties, the fruits with highest
content in bioactive compounds that expressed the best antioxidant activity.
INTRODUCTION
Mango (Mangifera indica L.) from Anacardiaceae Family, grows up to 1200 m high, with a high fruit production
is a tree 15 to 30 m high, with quick growth in shallow within 3 months of the year in low rain seasons (Bally,
and well drained soils, and pH oscillating from 5.5 to 7.5 2006). In Colombia, mango has more than 200 ecotypes
(Shah et al., 2010). In tropical zones of Colombia, it or genetically differentiated cultivars. However, there is
Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution
License 4.0 International License
no much information on its properties as functional food. In Colombia, mango is consumed mostly fresh and it is
In other hand, mango in currently known as one of the used in pulps, nectar, juices, jellies and jams elaboration
most important tropical fruit. It has been cultivated since (Mora et al., 2002). Due to the low agro-industrial
prehistoric times and its tree has been object of high development, sale in fresh is privileged. However, post-
veneration in India. Different countries cultivate this fruit, harvest losses reach approximately 40% of annual
among them are, Indonesia, Florida, Hawaii, Mexico, production (Sumaya-Martinez et al., 2012).
South Africa, Egypt, Israel, Brazil, Cuba and Philippines. Mango is one of the most consumed tropical fruit in the
Probably, India has more commercial plantations than the world, especially for its flavor, texture and versatility
rest of the world. However, mango’s economic (Oliveira et al., 2016). However, the studies that have
importance is due to the great local consumption in characterized its nutraceutical power in Caribbean
Caribbean countries, especially in Colombia (Michel et Colombian zone cultivars are few. That is why the
al., 2000). present work wants to know the metabolites content,
The consumption of fruits rich in biologically active specifically Mangiferin, carotenoids and total polyphenols,
compounds brings important benefits for human health; associated with antioxidant activity in six mango cultivars
mango is considered a fruit with nutraceutical properties from Córdoba, Colombia.
due to presence of carotenoids, ascorbic acid, fiber,
polyphenols, among others; which are associated to
antioxidant and anti-inflammatory properties, important in MATERIALS AND METHODS
treatments of metabolic disorders such as obesity and
Reagents and equipment
diabetes (Ibarra-Garza et al., 2015; Septembre-Malaterre
et al., 2016). Also, knowledge of concentration of Formic acid (p.a.), iron trichloride, sodium carbonate and Follin-
bioactive structures in fruits, allows obtaining information Ciocalteu reagent were obtained from Merck (Germany). Methanol
to implement extraction and drying techniques to give it and other solvents were acquired from Fisher Scientific Co. (Fair
greater added value as a functional food and to compete Lawn, NJ, USA); 2,2’-Azinobis(2-aminopropane) hydrochloride
in the international market with high quality products. (AAPH), fluorescein, Trolox®, 2,4,6-tri-(2-pyridyl)triazine (TPTZ)
were bought to Sigma-Aldrich Chem.Co (Millwakee, WI). Ultraviolet-
One of the bioactive compounds of greatest interest in visible measurements were done in a spectrophotometer Multiskan
the species M. indica is Mangiferin, a polyphenol type Spectrum (Thermo Scientific). The decrease in the intensity of
glycosyl xanthone that presents pharmacological actions fluorescein measured in ORAC assay was done in a
with antioxidant, anti-inflammatory, and neuroprotective spectrofluorimeter Perkin-Elmer LS-55, (U.K.). Chromatographic
properties (Takeda et al., 2007). Studies have suggested assays were done in a liquid chromatographer Shimadzu®,
that Mangiferin also has an in vitro anticancer effect in Prominence® UFLC series.
cellular lines of acute myeloid leukemia (AML) (Shoji et
al., 2011). Matkowski et al. (2013) reported Mangiferin as Vegetal material
a promising natural product with benefits as analgesic,
Vegetal material was collected in the Department of Córdoba,
antidiabetic, anti-sclerotic, antimicrobial, antiviral, cardio Colombia, especially in Monteria’s local market (18 M. ASL and a
and hepatoprotective, anti-allergic, monoamine mean temperature of 28°C in June 2016). Specimens in optimum
oxygenase (MAO) inhibitor, and protector against UV conditions of each variety were collected randomly in mature state.
radiation. After being stored in perforated polypropylene bags, they were
Carotenoids are pigments found in fruits and taken to the Laboratory Food Science, National University of
vegetables such as mango. Chemically, carotenoids are Colombia, Medellin, for analysis.
classified as tetraterpenes, of which more than 600
natural structures of different size, shape and polarity Sample preparation
have been identified. Carotenoids are associated with
various biological processes like antioxidant defense, 3.0000 g of pulp were homogenized with 20 ml of deionized water
photosynthesis and, some of them, as precursors of in Ultra-Turrax (IKA-WERK©) homogenizer. The extract obtained
from that was centrifuged at 5000 rpm for 10 min at room
vitamin A. Also, they determine the orange and yellow temperature. The supernatant recovered was stored at 4°C until the
colors of mango fruits. Many studies reported that an analysis.
adequate intake of carotenoids is related to the decrease
of various types of cancer (Melendez-Martínez et al.,
2007). Color determination
In other hand, condensed tannins are polymers formed The color was measured with a colorimeter Minolta (Minolta Co.
by flavan-3-ol units. Also, they are one of the principal Ltd., Osaka, Japan) on the basis of the color system CIELAB (L*,
antioxidant metabolites in products like tee and cocoa a*, b*). In this system L*, a* and b* describe a tridimensional space,
which content can be up to 35% of total polyphenols. where L* is the vertical axis and its value ranges from 100 for
Tannins are associated to the astringent flavor of fruits perfect white to zero for black. Values a* and b* specify the axis
green-red and yellow-blue, respectively. That values range from -60
and enzymatic browning reactions in mango bark to +60 or from -a (green) to +a (red) and from -b (blue) to +b
(Pierson et al., 2014). (yellow). To determinate the color, a portion of one side of each fruit
was obtained and measured longitudinally in three equidistant Carotenoids determination
points (Garza et al., 1996).
It was done by Ultraviolet-Visible (UV-Vis) spectrophotometry. In a
test tube, 1.0000 g of sample was added to 5.0 mL of cold acetone
Assessment of antioxidant capacity and it was left to stand for nearly 15 min in refrigeration (4°C). The
mixture was centrifuged at 1370 gravities for 10 min, the
Assessment of reducing power by FRAP method supernatant was collected in another test tube. The pellet was re-
extracted with 5.0 mL of cold acetone. Both acetone extracts were
This method assesses the reducing power of a sample according to mixed, and then, were filtered in Whatman paper No. 42 and its
its capacity to reduce ferric iron (Fe+3) with TPTZ to its ferrous form absorbance was determined at 449 nm. Carotenoid concentration
(Fe+2), which has its maximum absorbance at 593 nm, according to was obtained by the respective calibration curve, with β-carotene as
the design done by Benzie and Strain (1996). 50 µL of sample were pattern substance. Data was obtained by the Software SkanIt 2.4.2
added to 900 µL of a FRAP solution, acetate buffer pH 3.4, TPTZ RE for Multiskan Spectrum (Biswas et al., 2011). The results were
and FeCl3 in relation 10:1:1. The results were expressed as expressed as mg of β-carotene equivalent/100 g dry weight.
ascorbic acid equivalent antioxidant capacity (AEAC: mg of
ascorbic acid/100 g dry weight).
Ascorbic acid determination
Oxygen radical absorbance capacity (ORAC) assay It was established by HPLC. The aqueous supernatant was filtered
(pore size 0.45 µm) and dilutions in super pure water, before
The method described by Prior et al. (2005) and Romero et al. injection to chromatographer. A liquid chromatographer Shimadzu®
(2010) was used. 30 µL of sample were added to 21 µL of model LC-20AD was used, equipped with an auto-injector SIL-
fluorescein 1x10-2 M in PBS (75 mM), 2.899 µL of PBS (75 mM), 20A/HT, a communication module CBM-20A and a (PDA) SPD-
and 50 µL of AAPH 0.6 M in PBS (75 mM), temperature was M20A, calibrated to 245 nm. Ascorbic acid quantification was done
controlled to 37°C and pH was kept at 7.4. The readings were done with a C-8 (5 µm, 250 mm x 4.6 mm) column. Formic acid 0.1%
at an excitation ʎ 493 nm and excitation slit 10 nm. It was compared was used as mobile phase, at a flow rate of 0.8 mL min-1, at 35°C in
to the primary pattern Trolox® curve. The results were expressed isocratic conditions (Kelebek et al., 2009). The results were
as TEAC, µmol trolox equivalent /100 g of dry weight according to expressed as mg of ascorbic acid/100 g dry weight.
Equation 1.
L a b
Criollo 66 16 50
Zapote 56 25 50
Azúcar 76 7 50
Corazón 72 26 46
Jobo 76 19 43
Chancleta 66 16 50
phenols found in mango are chlorogenic, gallic, vanillin (Rungpichayapichet et al., 2015). The above, suggests
and protocatechuic acid, in order of abundance (Wall- Colombian mango as an important source of these
Medrano et al., 2014). On the other hand, in the mango’s pigments, not only for fresh consumption, but for its use
peel and seed, values between 3.8 and 13 GAE/100 g as a colorant additive in food matrices.
dry matter have been found (Dorta et al., 2012). Chong et
al. (2013) reported values between 120.70 and 210.24
mg gallic acid/100 g for mango cultivated in Malaysia. Vitamin C
Also, Lobo et al. (2017) showed that there is a great
oscillation in total phenol content in the Tommy Atkins Mango is recognized for its important contribution in
mango pulp cultivated in Brazil (46.18 – 116.93 mg gallic vitamins like ascorbic acid, thiamin, riboflavin, and niacin.
acid/100 g). Thanaraj et al. (2009_ reported 1055 and In this study, the varieties with the most vitamin C content
1691 mg gallic acid/100 g dry weight as the interval for were Chancleta and Jobo. However, the reported content
five mango varieties cultivated in Sri Lanka. A high for the six cultivars was lower than the one for other fruits
variability in total phenol content was reported for mango, studied in countries like Ecuador, Brazil and Colombia
due to the different weather and growing conditions of (Contreras-Calderón et al., 2011; Vasco et al., 2008).
each region. However, the evaluated Colombian pulps Seok et al. (2010) found that ascorbic acid content in
presented a significant content of phenolic compound as fresh mango from Malaysia was 136.8 mg/100 g dry
compared to what was reported by most authors. matter, a value close to the one found for Chancleta. The
variations in vitamin C content are due to the fact that this
compound is cataloged as the most unstable nutrient and
Total carotenoids prone to immediate loss after harvest. Therefore, its
content depends on the changes during the post-harvest
Carotenoids are the compounds responsible for handling, storage conditions, transformation and
coloration in most food, some of them are provitamin A, elaboration (Spínola et al., 2013).
like α and β-carotene, and β-cryptoxanthin. Recent
studies have made these pigments’ antioxidant properties
manifest, as well as their effectiveness in the prevention Condensed tannins
of certain human diseases, such as atherosclerosis or
even cancer (Müller et al., 2011). Polyphenols present in fruits and vegetables can include
Criollo and Jobo varieties presented the higher total simple structured compounds (phenolic acids), oligomers
carotenoid content with 105.88 and 196.15 mg β- (flavonoids, xanthones, stilbenes), or polymers
carotene equivalent/100 g dry matter, respectively. Silva (condensed tannins) (Pierson et al., 2014). Condensed
et al. (2014), who quantified bioactive compounds in tannins are oligomeric and polymeric proanthocyanidins
different fruit from Brazil, found 0.954 mg β-carotene that are widely distributed in the vegetable kingdom. It
equivalent/100 g dry matter in mango; Seok et al. (2010) has been reported that they are used to stop small local
reported that the carotenoid content in Malaysia’s mango bleeding, decrease buccal cavity inflammations, colds,
pulp was 0.650 equivalent β-carotene/100 g dry matter, bronchitis, burns, hemorrhoids, etc. (Piovesan et al.,
which are much lower values than the ones found in this 2017; Kasay et al., 2013). Also, the condensed tannins’
study. It is worth noting that there are 17 important chemical nature makes them a natural source of organic
carotenoids found in mango, among them β- compounds, with an application potentially wide for
cryptoxanthin, zeaxanthin, isomers of luteoxanthin, medicinal and industrial uses (Aguilar-López et al., 2012).
violaxanthin, neoxanthin and β-carotene, which is the last According to the results in Table 2, all mango fruits
one, the one with the highest prevalence presented a high condensed tannin content, highlighting
the varieties Corazón and Criollo. Gorinstein et al. (2011) FRAP values found in the evaluated mango pulps,
evaluated the bioactive compound content in different match with low quantity of vitamin C analyzed by HPLC.
exotic fruits, reporting 27.0 mg of catechin equivalent/100 However, ORAC unities from Chancleta, Criollo and Jobo
g dry weight as the condensed tannin content in mango. varieties are higher than the ones reported by Singh et al.
Arogba (2014) reported a content of 135.0 mg of catechin (2015) in dried Tommy mango chunks (408 to 651 µmol
equivalent/100 g dry weight in mango seed. The above TE/100 g dry sample). In other hand, ORAC values
shows that the evaluated mango varieties presented an between 0.46 and 1.33 mmol Trolox/100 g dry simple
important condensed tannin content, compounds whose were found in a study that was done on 10 cultivars of
antioxidant activity has been widely reported (Dobrecky Mediterranean diet fruits (Wojdylo et al., 2016). This
et al., 2014; Ocampo et al., 2014). shows that the analyzed varieties in this research exhibit
a considerable antioxidant capacity measured with ORAC
methodology. It is worth to mention that this fluorometric
Mangiferin technique is the one endorsed by the United States
Department of Agriculture (USDA) for measuring
Mangiferin is the main polyphenol that constitutes antioxidant capacity in food and nutritional supplements
mango’s leaves, fruit and bark. The literature has (Rojano et al., 2012).
reported its wide pharmacological, antidiabetic, antitumor,
immunomodulatory and antioxidant activity (López et al.,
2015). According to the results presented in Table 2, Correlations between polyphenols and antioxidant
mangos cultivated in Córdoba present similar values and activity
even higher for Mangiferin content reported in different
research. For example, Luo et al. (2012) reported that the As shown in Figures 1 and 2, a positive correlation was
Mangiferin content in 11 varieties of mango pulp from found between total phenol content and antioxidant
China, was between 0.2 and 20 mg/100 g dry matter. capacity evaluated with ORAC and FRAP (r2 = 0.87 and
Other authors suggested that Mangiferin is mainly found r2 = 0.84), which shows that polyphenols are the main
in mango’s peel, leaves and bark, and in smaller contributors to mango’s antioxidant activity. Also, the
measure, in pulp, reporting values from 169 mg/100 g dry synergism between antioxidants could explain why the
matter in peel, and 4.2 mg/100 g dry matter in seeds fruit’s antioxidant capacity is higher than the individual
(Rymbai et al., 2015). This shows that the evaluated content in each antioxidant, like vitamin C. Similar results
Colombian mangos have a promising Mangiferin content, have been found in other researches (Thaipong et al.,
highlighting their nutraceutical quality and encouraging 2006; Silva and Sirasa, 2016).
industrialization of this fruit in functional products. The
high Mangiferin content in the studied varieties can be
explained since this xanthone is synthesized by the Conclusion
phenylpropanoid pathway in presence of high solar
The present study showed the antioxidant potential of six
radiation, which is a representative condition of the
mango varieties cultivated in the Department of Córdoba,
Caribbean Colombian region (Ruiz and Romero, 2001).
being Chancleta, Criollo and Jobo, the varieties that
showed the highest antioxidant activity in vitro. As
regards mangiferin content, all varieties stood out above
Antioxidant capacity
fruits from other countries. These results prove that
Colombian mangoes have a high nutraceutical potential
FRAP and ORAC-Total
due to its Mangiferin content and to the antioxidant
expression of the other bioactive compounds like
Different methodologies have been proposed to evaluate
carotenoids and tannins. It is necessary to conduct
fruit’s antioxidant capacity, of which ferric ion reducing
studies in vivo to determine bio-accessibility and/or bio-
antioxidant power (FRAP) and oxygen radical
availability of mango’s extracts with higher antioxidant
absorbance capacity (ORAC) methods are widely used.
potential and Mangiferin as the main bioactive.
These methods measure different mechanisms of
antioxidant activity; while ORAC is related to capacity for
neutralizing a free radical using HAT mechanism, FRAP CONFLICT OF INTERESTS
assay measures sample’s capacity for reducing the ion
Fe3+ to Fe2+ (SET mechanism) (Rodríguez et al., 2010; The authors declare there is no conflict of interest.
Botero et al., 2007). In this research, the variety with the
best reducing capacity was Chancleta with 296.06 mg of
ascorbic acid/100 g dry sample, followed by Criollo and ACKNOWLEDGEMENTS
Azúcar. In general, low values of reducing capacity have
been reported for mango (Paz et al., 2015). The authors express their gratitude to Colciencias for its
2500
1500
1000
R² = 0.8701
500
0
0 50 100 150 200 250 300
orac-total (µmol Trolox/100 g DB)
350
300
250
200
150
100
50 R² = 0.84
0
0 50 100 150 200 250 300
FRAP (mg ascorbic acid/100 g DB)
support through the fellowship-internship Young Arogba SS (2014). Phenolics, Antiradical Assay and Cytotoxicity of
Processed Mango (Mangifera indica) and Bush Mango (Irvingia
Researchers and Innovators (Call 761)”, the entities in
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of the Pacific Alliance (VIII Call) -ICETEX, APC Evolution of pigments and their relationship with skin color based on
(Presidential Agency of International Cooperation of ripening in fruits of different Moroccan genotypes of apricots (Prunus
armeniaca L.). Sci. Horticult. 207:168-175.
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Bally ISE (2006). Mangifera indica (mango), ver 3.1 In: Elevitch, C.R.
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