Aronia melanocarpa berries: Phenolics composition and antioxidant properties changes during fruit development and ripening

Abstract

Aronia melanocarpa E. (black chokeberry, aronia) is cultivated in Poland, the USA, Korea and many other countries worldwide. It is known that its ripening and ripe berries are a very rich source of polyphenolic antioxidants, however, there is no data concerning unripe fruits. In this work, the changes in the content of anthocyanins, procyanidins, total polyphenols, flavonoids and antioxidant activity (measured with ORAC and DPPH-EPR tests) of Aronia melanocarpa E., Nero cultivar, during the whole fruit development and ripening period were studied. The highest content of total polyphenols (up to 20 g/100 g d.w.), procyanidins (10-15 g/100 g d.w.) and flavonoids (7-11 g/100 g d.w.) as well as the highest antioxidant activity (up to 100 mmol Trolox/100 g d.w.) was observed for unripe fruits. Procyanidins content declined during fruit development and then increased slightly in later maturation stages. Anthocyanin biosynthesis initiated at the beginning of fruit ripening and reached the highest level (2-3 g/100g d.w.) in mature fruit. Thus, although as for now only ripe berries are processed to obtain juice and extracts for foods, our results suggest that green berries rich in procyanidins and other phenolics may be an interesting raw plant material for both food and pharmaceutical industries.

Full text

214 Emir. J. Food Agric ● Vol 31 ● Issue 3 ● 2019
Aronia melanocarpa berries: phenolics composition and
antioxidant properties changes during fruit development
and ripening
Małgorzata Gralec, Iwona Wawer, Katarzyna Zawada*
Department of Physical Chemistry, Faculty of Pharmacy with the Laboratory Medicine Division, Medical University of Warsaw, Banacha 1 Str.,
PL02097 Warsaw, Poland
INTRODUCTION
Though Aronia melanocarpa E. (black chokeberry, aronia)
is native to eastern North America, it is cultivated
extensively in Europe and in Asia. The aronia fruit is
a very rich source of dietary antioxidants (Oszmiański
and Wojdyło, 2005; Kulling and Rawel, 2008). Ripe
A. melanocarpa berries contain various types of compounds:
anthocyanins, procyanidins and flavonols (quercetin
glycosides) (Wu et al., 2004; Slimestad et al., 2005). These
polyphenolic components of fruits make them a valuable
material which can be used as food or food supplements
dedicated to protect from oxidative stress (Battino et al.,
2009). On the other hand, it has been suggested that
also unripe fruits can be a valuable material with better
antioxidant properties than mature fruits (Wang and Lin,
2000; Castrejón et al., 2008, Tulipani et al., 2011). However,
although the properties and chemical composition of ripe
aronia fruits are well known, green aronia fruits have not
been studied so far. Green aronia fruits are sour and bitter
and are not suitable for direct human consumption. Still,
procyanidins, avonoids and other secondary metabolites
can be benecial for human health when extracted from
fruits.
The consumption of A. melanocarpa berries could have
a positive impact on human health (Banjari et al., 2017;
Chrubasik et al., 2010; Gawryś et al., 2012; Jurikova et al.,
2017; Kokotkiewicz et al., 2010; Kulling, 2008). Numerous
studies indicated that aronia extracts from ripe fruits have
anticancerous (Gasiorowski et al., 1997; Malik et al., 2003),
antidiabetic (Simeonov et al., 2002, Baum et al., 2016) and
anti-inammatory (Ryszawa et al., 2006) properties, reduce
blood pressure (Bell and Gochenaur, 2006) and alleviate the
toxicity of heavy metals (Kowalczyk et al., 2003).
Phenolic compounds and the antiradical activity of different
cultivars of aronia were compared during two consecutive
years (Jakobek et al., 2012). Although the profile of
Aronia melanocarpa E. (black chokeberry, aronia) is cultivated in Poland, the USA, Korea and many other countries worldwide. It is known
that its ripening and ripe berries are a very rich source of polyphenolic antioxidants, however, there is no data concerning unripe fruits. In
this work, the changes in the content of anthocyanins, procyanidins, total polyphenols, avonoids and antioxidant activity (measured with
ORAC and DPPH-EPR tests) of Aronia melanocarpa E., Nero cultivar, during the whole fruit development and ripening period were studied.
The highest content of total polyphenols (up to 20 g/100 g d.w.), procyanidins (10-15 g/100 g d.w.) and avonoids (7-11 g/100 g d.w.) as
well as the highest antioxidant activity (up to 100 mmol Trolox/100 g d.w.) was observed for unripe fruits. Procyanidins content declined
during fruit development and then increased slightly in later maturation stages. Anthocyanin biosynthesis initiated at the beginning of
fruit ripening and reached the highest level (2-3 g/100g d.w.) in mature fruit. Thus, although as for now only ripe berries are processed to
obtain juice and extracts for foods, our results suggest that green berries rich in procyanidins and other phenolics may be an interesting
raw plant material for both food and pharmaceutical industries.
Keywords: Aronia melanocarpa; Chokeberry; Antioxidants; Polyphenols; Anthocyanins; Green fruit
ABSTRACT
Emirates Journal of Food and Agriculture. 2019. 31(3): 214-221
doi: 10.9755/ejfa.2019.v31.i3.1921
http://www.ejfa.me/
RESEARCH ARTICLE
*Corresponding author:
Katarzyna Zawada, Department of Physical Chemistry, Faculty of Pharmacy with the Laboratory Medicine Division, Medical University of
Warsaw, Banacha 1 Str., PL02097 Warsaw, Poland . Tel.: +(48) 225720950, E-mail: katarzyna.zawada@wum.edu.pl
Received: 21 January 2019; Accepted: 30 March 2019

Gralec, et al.
Emir. J. Food Agric ● Vol 31 ● Issue 3 ● 2019 215
polyphenols was the same, some differences were found
in the content of these compounds. Such differences were
also observed by Howard et al. (2003) for various cultivars
of blueberries and by Lestario et al. (2017) for Java plum.
The present research work describes the major chemical
changes and antioxidant activity of Polish cultivar “Nero”
during three seasons of fruit development. The aim of the
study was the determination of the optimal collection time
in order to get aronia berries rich in particular groups of
polyphenolic compounds.
Although there are available numerous studies on the
changes in bioactive compounds during aronia maturation
process (Jeppsson and Johansson, 2000; Banjari et al., 2015;
Bolling et al., 2015), there is no information concerning the
rst stages of development, i.e., unripe fruits. Thus, the
composition of phenolic compounds in fruits in different
development stage (i.e., unripe, ripening and ripe) was
examined. Since the biological activity of A. melanocarpa
components is usually attributed to their antioxidant
properties, these characteristics of aronia berries in
different stages of development were studied as well.
Many assays are used for estimating antioxidant activity
of a vegetable or fruit matrix. DPPH and ORAC are
the two most common tests, and have been used to
measure the antioxidant activity of e.g. sweet orange juice
(Giuffrè et al., 2017a), coffee (Yashin et al., 2013), guava
fruit extracts (Thaipong et al., 2006) as well as edible
vegetable oils (Giuffrè et al., 2016; Giuffrè et al., 2017b).
Therefore, DPPH and ORAC tests were chosen, as they
are the assays most often applied to aronia fruit as well, to
enable comparison with earlier studies on the antioxidant
properties of berries.
MATERIALS AND METHODS
Plant material
Fruits of Aronia melanocarpa E. (“Nero” cultivar) were
collected from May to August in 2012, 2013 and 2016 at
the plantation in Mazowieckie District, Poland. The plants
were six years old. The local climate is warm-summer
humid continental climate, “Dfb” according to the
Kppen classication. The average annual temperature and
precipitation of the region are of 7.5°C and 550–600 mm,
respectively. The plants were rain-fed only (no irrigation).
The fertilization pattern was the same in all studied years,
mineral fertilization only, applied in May.29 May was
arbitrarily taken as the rst day of fruit development
(day 0). Collected fruits were frozen and stored at -15oC.
They were classied according to their color as unripe
(green fruit), ripening (pink tinted green to red fruit) and
ripe (purple-black fruit).
Sample preparation
The extraction procedure was based on that proposed
by Oszmiański and Wojdyło (2005). Frozen chokeberries
were lyophilized and ground. Three independent batches
from every sample (described by date of collection, 1 g
each) were extracted with methanol acidied with 1 g/kg
HCl. The extraction was performed by sonication for
20 minutes at room temperature. After that, the samples
were centrifuged and supernatants were used for further
analysis of antioxidant activity, as well as the content
of anthocyanins, procyanidins and flavonoids. Some
studies were performed immediately after the collection
of supernatants, and for other analyses the samples were
stored at -30oC for a maximum of two weeks.
Analytical procedures
Total anthocyanins content
The content of anthocyanin was determined using the
pH-differential method (Giusti and Wrolstad, 2001) using
Evolution 60S spectrophotometer (Thermo-Fisher, USA).
The content of anthocyanin pigment was calculated
for each extract and expressed as cyanidin-3-glucoside
equivalent (C3GE) in g/100 g d.w. of fruits.
Total avonoids content
The method proposed by Christ and Müller (1960)
was used to determine the content of avonoids. The
absorbance at 510 nm was measured using Evolution 60S
spectrophotometer (Thermo-Fisher, USA). The results
were expressed as catechin equivalent (CE (g/100 g d.w.)).
Total procyanidins content
Procyanidin content was determined with vanillin method
and the parameters were based on the study of Sun et al.
(1998). The absorbance was measured at 500 nm by Evolution
60S spectrophotometer (Thermo-Fisher, USA). Results were
expressed as epicatechin equivalents (EE (g/100g d.w.)).
Total phenolics content (TP)
Folin-Ciocalteu colorimetric method (Singleton et al., 1999)
was used to determine total phenolics. Briey, 20 μl of a
diluted extract samples were placed in each microplate’s
well and then 100 μl of Folin-Ciocalteu reagent (0.3 mol/L)
was added to each well. The mixture was kept in the dark
for 5 minutes at room temperature. Then 50 μl of 200 g/L
sodium carbonate was added. The mixture was incubated
for 20 minutes at 37°C. The absorbance at 765 nm was
measured using microplate reader Synergy Mx (Biotec,
USA). The results were expressed as gallic acid equivalents
(GAE (mg/100g d.w.)).
DPPH-EPR radical scavenging assay
The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical
scavenging test was used to determine the antioxidant

Gralec, et al.
216 Emir. J. Food Agric ● Vol 31 ● Issue 3 ● 2019
activity (Sanna et al., 2012). The extracts are coloured
and/or cloudy, thus for popular spectrophotometric
UV-vis measurements the background corrections for
absorbance are necessary. Therefore, radical scavenging
activity was estimated by electron paramagnetic resonance
(EPR) spectroscopy. EPR spectra were measured at
ambient temperature (298 K) on a MiniScope MS200
EPR spectrometer (Magnettech, Germany). The samples
were diluted with methanol (30-100 fold). Equal volumes
of a diluted extract and of DPPH methanolic solution
(3.4 mmol/L) were mixed, and after 20 minutes EPR
spectra were taken. The intensity of registered EPR spectra
was compared with the control sample (methanol in
place of an extract). The results were expressed as Trolox
equivalents (TE) in mmol TE and recalculated for 100 g d.w.
ORAC-uorescein (ORAC-FL) assay
The method is based on that proposed by Ou et al. (2001).
All solutions were prepared in PBS (phosphate-buffered
saline), pH 7.4. For measurements, 30 μl of the chokeberry
extract diluted with PBS (100-500 fold), standard (Trolox)
solution or, in case of a blank, 30μl of PBS buffer, were
mixed with 180 μl of 112 nmol/L uorescein solution in
a well of 96-well plate and thermostated for 15 minutes
at 37°C. Then, 100 μl of 100 mmol/L AAPH solution
was added. Fluorescence was measured with F-7000
Fluorescence Spectrophotometer (Hitachi, Japan) equipped
with a Micro Plate Reader every 70 s for 90 minutes. The
reaction mixture was thermostated at 37°C. ORAC values
expressed as Trolox equivalents (TE (mmol/100g d.w.))
were calculated using the standard curve.
HPLC/MS characterization of fruit contents
Characterization of contents of methanolic extracts from
unripe, ripening and ripe aronia fruit was performed using
Ultra-Performance Liquid Chromatograph ACQUITY
UPLC I-Class (Waters Inc) coupled with Synapt G2-S
HDMS (Waters Inc) mass spectrometer equipped with an
electrospray ion source and q-TOF type mass analyzer.
The ACQUITY UPLCR BEH C18 1.7um (WATERS)
column was used. The mobile phase consisted of 0.1%
formic acid in water (solvent A) and methanol (solvent B)
with the following gradient conditions: 95% A at 0–2 min,
95–0% A at 2–15 min, 0% A at 15-22 min, 0–95% A at
22–22.10 min and 95% A at 22.10–25 min. The ow rate
was 0.3 mL/min.
Statistical analyses
Statistical analysis (one-way ANOVA, correlation analysis)
was performed with Statistica 10 (StatSoft Inc.) software.
The Scheffe test was applied to assess signicant differences
(p < 0.05) between samples.
RESULTS AND DISCUSSION
The content of bioactive compounds
The content of the main groups of compounds:
anthocyanins, avonoids and procyanidins, as well as the
content of total phenolics, change in time during fruit
development and ripening. The results are shown in Fig. 1.
At the beginning of fruit development, the berries are
green and do not contain anthocyanins. These compounds
appear later as the fruits are ripening. An increase in the
anthocyanin pigment concentration was observed in aronia
fruits between 30th and 80th day of fruit development
(Fig. 1a). The content of anthocyanins increases from
about 1 g/100 g of dry weight in the last days of July
(50th - 55th day of fruits development) to 2-3 g/100 g
Fig 1. Content of (a) anthocyanins, expressed as cyanidin-3-glucoside equivalent (C3GE) in g/100 g d.w., (b) avonoids in aronia berries,
expressed as catechin equivalent (CE [g/100 g d.w.]), (c) procyanidins in aronia berries, expressed as epicatechin equivalents (EE [g/100g d.w.]),
(d) total phenolics in aronia berries, expressed as gallic acid equivalents (GAE [mg/100g d.w.] in aronia berries as the function of time during
three vegetation seasons. Error bars correspond to standard deviation of three repetitions. Samples with the same letters are not signicantly
different (one-way ANOVA, Sheffe test, p>0.05).

Gralec, et al.
Emir. J. Food Agric ● Vol 31 ● Issue 3 ● 2019 217
in August (80th - 90th day). The highest concentration
of anthocyanins was recorded in ca. 80 day-old fruit.
A slight, 10%, decrease in 2013 observed at the end of
collection period may be due to a decrease in acidity. It was
observed (Holcroft and Kader, 1999) that the stability of
anthocyanins as well as their biosynthesis could decrease
with the decrease in acidity. This tendency of changes in
anthocyanin content is similar to that observed by other
authors (Jeppsson and Johansson, 2000; Andrzejewska
et al., 2015; Bolling et al., 2015). It should be noted, though,
that in previous studies the fruits were collected only during
the ripening period and later (August - October).
The tendency of changes in anthocyanin content is similar in
all three vegetation seasons, but fruits collected in 2016 have
lower nal content of anthocyanins than those harvested
in previous years. The differences in concentrations and
the progress of ripening could be tentatively connected
with weather, as the vegetation season of 2016 was
characterized by lower, than in previous years, temperature
in August (the average temperature in this month in 2012
was 18.9oC, in 2013 19.3oC and in 2016 16.9oC (https://
www.wunderground.com/history/). Weather conditions,
such as temperature and insolation, inuenced phenolics
content in juice of aronia, as shown recently (Tolić et al.,
2017). As there was no change in irrigation or fertilization
pattern among years, these differences cannot be caused
by the variation of cultivation conditions.
The analysis of anthocyanins content conrmed that ripe
Aronia melanocarpa fruit is a rich source of these phenolics
(2-3 g/100 g d.w.). It is consistent with the ndings of
Oszmiański and Wojdyło (2005) (2 g/100 g d.w. of ripe
aronia fruit) as well as of Kapci et al. (2013) (about
2 g/100 g d.w., assuming the average water content of
ca. 800 g/kg of fresh fruit). Aronia fruit contains more
anthocyanins than blueberry (about two times more), açaí
(about four times more) and goji (even 350 times more)
(de Moura et al., 2018).
The highest level of avonoids, determined by Christ and
Müller method, was observed in unripe fruits. It amounts
from 7 g/100 g d.w. in 2012 and 2013 up to 11 g/100 g d.w.
in 2016. The content of these compounds is declining to
about 4 g/100 g d.w. as fruits are ripening, as illustrated in
Fig. 1b. It stays at the same level in August, until the end of
observation. More rapid decrease of avonoid content is
observed in case of fruits collected in 2016 than for these
collected in 2012 and 2013. The results of our analysis are
similar to the ndings of Oszmiański and Wojdyło (2005)
(2.1 g/100 g d.w. of all analysed by HPLC avonoids,
i.e., sum of anthocyanins and quercetin glycosides). Kapci
et al. (2013), using the Christ-Müller method, obtained
values of 1.99 and 1.25 g/100 g for dried chokeberry fruits.
It is worth mentioning that in the conventional oven-drying
of fruit there is usually a larger decrease in avonoid
content than during freeze-drying (Thi and Hwang, 2016)
which was used in our work. It could be the reason of
lower values of Kapci et al. (2013). As Bolling et al. (2015)
analyzed the bioactive compounds in juice obtained from
aronia fruits harvested at different time points during the
ripeness period (August – September), it is difcult to
compare values of avonoid content. Still, in their work no
apparent trend of changes by harvest date was observed,
similarly to our work.
Fig 2. Correlation between: (a) total phenolics content and procyanidin content (the correlation parameters: r = 0.81, p < 0.05), (b) the results of
DPPH test and total phenolics content (the correlation parameters: r = 0.70, p < 0.05), (c) the results of ORAC test and total avonoids content
(the correlation parameters: r = 0.77, p < 0.05) of aronia berries in different stages of development.

Gralec, et al.
218 Emir. J. Food Agric ● Vol 31 ● Issue 3 ● 2019
Similar situation to the one with avonoids is observed in
the case of procyanidins. Their concentration decreases
during maturation of fruits. The amount of these
compounds is within 10 g to 15 g/100 g d.w. in May and
June (0-28 day), for green fruits, and declines in August
(70-88 day) to about 8 g/100 g d.w. in 2012, 4-5 g/100 g d.w.
in 2013, and even to 1 g d.w. in 2016. It stays at this level
in ripe fruits (Fig. 1c). It is in agreement with the ndings
of Wu et al. (2004) (about 2.5 g in ripe aronia, assuming
800 g/kg water content of fresh fruit). The larger decrease
of procyanidin content is observed in fruits collected in
2013 and 2016 than in 2012. The content of procyanidins
observed by us during the whole ripeness period is contrary
to that noticed by Bolling et al. (2015), where this content,
determined with DMAC method, increased with time.
Aronia showed the highest total phenolics concentration
during the initial stage of fruit development; ca. 20 g/100 g
d.w. was recorded in 30 day-old fruit (Fig. 1d). It is a much
higher value than the one obtained for unripe blueberries
(4 – 7 g/100 g d.w.) (Castrejón et al., 2008) or thornless
blackberry (2 g/100 g d.w.) (Wang and Lin, 2000). Later,
there was a signicant reduction in total phenolics from
30th to 90th day of fruit development. A decrease in phenolic
compounds content with maturation and ripening has also
been reported in guava (Bashir and Abu-Goukh, 2003),
blueberry (Castrejón et al., 2008) and other fruits.
The positive correlation between total content of phenolic
compounds and procyanidins has been observed (illustrated
in Figure 2a). No significant correlation was observed
between other groups of active compounds present in the
berries. Thus, procyanidins are mainly responsible for the
changes in total phenolics content of A. melanocarpa fruits
during ripening. A weak correlation between total phenolic
compounds and procyanidins content was also observed by
Bolling et al. (2015) for ripe aronia berries collected from
August to September, which is in agreement with our ndings.
Although there was no correlation between total
anthocyanin and total polyphenol content when the
whole season has been taken into account, a decrease
in total phenolics (Fig. 1d) coincided with an increase in
anthocyanin pigment content (Fig. 1a). A similar effect was
observed for blueberries by Castrejón et al. (2008) during
fruit development and ripening. A decrease in the content
of total phenolics and especially procyanidins reduces the
astringency of fruit which is a desirable sensory attribute.
The increase of anthocyanins concentration enhances the
color and visual attractiveness of the fruit.
Antioxidant activity
Oxidative stress is suspected to be an important factor
in developing neurodegenerative (Patel and Chu, 2011)
and cardiovascular diseases (Csányi and Miller, 2014).
Consuming food rich in antioxidants could protect our
bodies from oxidative stress (Bjørklund and Chirumbolo,
2017). The radical-scavenging-linked antioxidant properties
of the extracts from black chokeberry cultivated in Korea
was investigated (Hwang et al., 2014). The results suggest
that black chokeberry extracts could be considered as a
good source of natural antioxidants and functional food
ingredients.
As the biological activity of A. melanocarpa berries is
often ascribed to their antioxidant properties, it seemed
worth checking the relation between the content of
active substances and antioxidant activity of fruits. The
antioxidant activity of aronia fruits collected at various
stages of development was determined with the DPPH-
EPR and the ORAC tests.
DPPH radical scavenging assay is the most popular
screening test for antioxidants. It is usually performed with
spectrophotometric monitoring of DPPH concentration.
It should be noted, though, that many food components
can interfere with such measurements as their absorption
spectra overlap with the DPPH radical UV-vis spectrum.
Furthermore, in polar solvents the aggregation of DPPH
could take place and inuence the results. Both problems
Fig 3. Antioxidant activity determined with (a) DPPH-EPR assay, (b)
ORAC assay, expressed as Trolox equivalents (TE [mmol/100g d.w.]),
as the function of time during three vegetation seasons. Error bars
correspond to standard deviation of three repetitions. Samples with the
same letters are not signicantly different (one-way ANOVA, Sheffe
test, p>0.05).

Gralec, et al.
Emir. J. Food Agric ● Vol 31 ● Issue 3 ● 2019 219
can be avoided with the use of EPR spectroscopy, as it is
sensitive only to substances with unpaired electrons and also
immediately shows the DPPH aggregation (Sanna et al., 2012).
ORAC assay is based on the ability of antioxidants present
in the sample to inhibit the oxidation of a probe by peroxyl
radicals. Recently, a move to widen the spectrum of reactive
oxygen and nitrogen species used in this test in order to
better predict the in vivo activity of a given antioxidants
source has been proposed (Prior et al., 2016).
The results of antioxidant activity determination are
presented in Fig. 3. Results of these tests indicate that
unripe fruits of A. melanocarpa collected at the beginning of
growth have higher antioxidant activity than the ripe ones.
There is a statistically signicant difference in antioxidant
activity of unripe and ripe aronia fruits. A similar
tendency in ORAC values was observed for other berries,
e.g. black raspberry or thornless blackberry (Wang and
Lin, 2000), however, the obtained values (16 and 18 mmol
Trolox/100 g d.w., respectively) were much lower than for
aronia (up to 100 mmol Trolox/100 g d.w.).
The antioxidant activity was similar, considering their values
as well as the tendency to decrease during fruit growth
and maturation, for fruits harvested in all three vegetation
seasons. The fruits collected at the end of May (day 0) and
at the beginning of June (days 3-6) exhibited the highest
antioxidant activity. Then it decreased signicantly during
July (days 32-63), when maturation took place, to reach
plateau in August (days 64-95).
Antioxidant properties of aronia determined with the DPPH
test positively correlate with the content of total polyphenols
both for ripe and unripe fruit, as illustrated in Fig. 2b, while
the results of ORAC test positively correlate with the
content of total avonoids, as illustrated in Fig. 2c. It may
be explained by different mechanisms of these antioxidant
assays. There is no signicant correlation between antioxidant
properties determined by any of the assays and the content
of anthocyanins. Thus, it suggests that the antioxidant activity
of A. melanocarpa berry extracts is due to different groups of
bioactive compounds than anthocyanins.
HPLC/MS analysis
According to the HPLC chromatograms and the mass
spectrometry results, the main anthocyanin in aronia
fruit was cyanidin-3-galactoside, which was detected in
ripening and ripe fruit (Table 1). The presence of second
anthocyanin, cyanidin 3-arabinoside was observed only in
ripe fruit, whereas the catechin was detected only for unripe
(green) fruit. The highest concentration of chlorogenic
acids and quercetin derivatives was determined for ripe
fruit, followed by unripe fruit. For ripening fruit the
temporary decrease in their concentration was observed.
CONCLUSIONS
The Aronia melanocarpa fruits are a rich source of phenolics.
Chemical composition of the berries signicantly changes
during fruit development and ripening. Collection of the
berries as a food and food supplements material should
take into account the changes in chemical components
during ripening. The green, unripe fruits have the highest
antioxidant activity due to the high content of procyanidins
and avonoids, in spite of the absence of anthocyanins.
Thus, the extracts of green fruits could be potentially used
as a dietary supplement in prophylactics as well as dietary
support of the therapies. The polyphenol-rich extracts may
be especially useful for patients with metabolic syndrome
where the severe oxidative stress occurs. In our opinion, the
extract of green berries, rich in polyphenols and exhibiting
strong antioxidant activity, could be an interesting new
material for dietary supplements and deserves further
studies. On the other hand, fruits harvested in August
are characterized by the highest content of anthocyanins,
and could be used as an important dietary component
for people with diabetes and as the support of therapy
of cardiovascular diseases. In summary, the current study
revealed that fruit development and ripening process
affects polyphenolic compounds composition and content.
Understanding the pattern of their concentration changes
can contribute to improving breeding and harvest strategies.
ACKNOWLEDGMENT
This work was supported by the Polish National Science
Center [grant number 2015/17/B/NZ7/03089].
Table 1. HPLC-MS identication of phenolic compounds in
A. melanocarpa fruit in different stages of development

Gralec, et al.
220 Emir. J. Food Agric ● Vol 31 ● Issue 3 ● 2019
Authors’ contributions
M.G. collected plant material, M.G. and K.Z. performed
the measurements, I.W. and K.Z. were involved in planning
and supervised the work, M.G. and K.Z. processed the
experimental data, performed the analysis, drafted the
manuscript and designed the gures. All authors discussed
the results and commented on the manuscript.
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