Food Chemistry 239 (2018) 1160–1166

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Food Chemistry

j o u r n a l h o m e p a g e : w w w . e l s e v ie r . c o m / l o c a te / f o o d c h e m

Effect of ascorbic acid postharvest treatment on enzymatic browning, phenolics

and antioxidant capacity of stored mung bean sprouts
⇑ ´
Małgorzata Sikora , Michał S wieca
Department of Biochemistry and Food Chemistry, University of Life Sciences, Skromna Str. 8, 20-704 Lublin, Poland

article info abstract

Article history: Enzymatic browning limits the postharvest life of minimally processed foods, thus the study selected the optimal inhibitors of
Received 14 February 2017 polyphenol oxidase (PPO) and evaluated their effect on enzymatic browning, phe-nolics and antioxidant capacity of stored
Received in revised form 11 July 2017 mung bean sprouts. The sprouts treated with 2 mM and 20 mM ascorbic acid had a lowered PPO activity; compared to the
Accepted 13 July 2017 Available online 14
control by 51% and 60%, respectively. The inhibition was reflected in a significant decrease in enzymatic browning. The
July 2017
sprouts treated with 20 mM ascorbic acid had 22% and 23% higher phenolic content after 3 and 7 days of storage,
respectively. Both storage and ascorbic acid treatment increased potential bioaccessibility of phenolics. Generally, there was
Keywords:
no effect of the treatments on the antioxidant capacity; however, a significant increase in the reducing potential was
Antioxidant capacity
determined for the sprouts washed with 20 mM ascorbic acid. In conclusion, ascorbic acid treatments may improve consumer
Ascorbic acid
Enzymatic browning quality of stored sprouts.
Mung bean
Polyphenol oxidase 2017 Elsevier Ltd. All rights reserved.
Sprouts

1. Introduction
Beans are consumed all over the world due to their high nutri-tional and
nutraceutical value and desirable taste (Peñas, Gómez, Frías, & Vidal-
Valverde, 2010). They are a good and inexpensive source of dietary proteins,
carbohydrates, minerals, vitamins (A, B, C, E), and antioxidants (Chandrasiri,
Liyanage, Vidanarachchi, Weththasinghe, & Jayawardana, 2016; Paja˛ k,
Socha, Gałkowska, Roznowski, & Fortuna, 2014). Mung bean is rich in
proteins (20– 33%) with amino acid composition comparable with soybean
being FAO/WHO reference protein (Mubarak, 2005; Peñas et al., 2010).
Additionally, mung bean contains bioactive components, including
flavonoids, phenolic acid, organic acid, amino acids, carbohydrates, and lipids
(Shi, Yao, Zhu, & Ren, 2016; Tang, Dong, Ren, Li, & He, 2014). Diet
enriched in mung bean has a positive influence on human health and exhibits
detoxifying, anti-inflammatory, cholesterol-lowering, and diuretic properties
(Peñas et al., 2010).
In recent years, sprouts have been recognized as functional food products
(Shi et al., 2016). During sprouting, storage materials (starch, fats, and
proteins) are decomposed into simple com-pounds, which are easily
bioaccessible. Meanwhile, phytic acid is degraded; thus, the bioavailability of
minerals is significantly increased. Germination has a positive effect on the
profile of antioxidant compounds as well (Shi et al., 2016). Compared to
⇑ Corresponding author.
E-mail address: malgorzata.sikora@up.lublin.pl (M. Sikora).
dormant seeds, mung bean sprouts have stronger antioxidant properties
mainly due to the higher content of polyphenols, i.e. compounds exhibiting a
number of properties beneficial to human health, including alleviation of
cardiovascular disease, inhibition and reduction of the risk of certain cancers,
and protection against diabetes, osteoporosis, and neurodegenerative diseases
(Cevallos-Casals & Cisneros-Zevallos, 2010; Pandey & Rizvi, 2009; Tang et
al., 2014).
Before consumption, sprouts are usually stored under cool con-ditions,
which is aimed to preserve microbiological purity and inhi-bit sprout
´
metabolism and growth (S wieca & Gawlik-Dziki, 2015). One of the main
problems influencing the consumer quality of low processed food (sprouts)
occurring during storage is enzymatic browning (Redondo, Venturini, Oria, &
Arias, 2016). Polyphenol oxidase (PPO) is the major cause of enzymatic
browning in higher plants (Chen et al., 2017). The reaction is a consequence
of the oxi-dation of phenolic compounds by PPO, which triggers the genera-
tion of dark pigments (Severini, Baiano, De Pilli, Romaniello, & Derossi,
2003). This is particularly relevant for legumes, which are rich in polyphenols
and highly susceptible to enzymatic browning, causing a decrease in the
polyphenol content and subse-quent lowering of nutraceutical quality
(Holzwarth, Wittig, Carle,
& Kammerer, 2013). There are many trials aimed to preserve food against
enzymatic browning (Kaaber, Martinsen, Bråthen, & Shomer, 2002). They
involve biological substances (e.g. whey protein concentrate (Altunkaya,
2011) and phytocides from pine leaves (Kim, Kim, Chung, & Moon, 2014)),
physical (pH,

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8146/ 2017 Elsevier Ltd. All rights reserved.
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M. Sikora, M. Swieca / Food Chemistry 239 (2018) 1160–1166
temperature, pressure treatment) (Zhan, Li, Hu, Pang, & Fan, 2012), and
chemical compounds (inhibitors of PPO activity) limiting access of oxygen
(Altunkaya & Gökmen, 2009). Between effective inhibitors of PPO activity,
citric acid, ascorbic acid, sulfur dioxide, and other compounds having thiol
groups (L-cysteine, glutathione) and peptides should be mentioned (Holzwarth
et al., 2013). Inhibi-tor compounds added to foods should be non-toxic and
highly active at low concentrations; they should not adversely affect the taste
and odor of stored food. They should also be resistant to tem-perature and
other technological parameters (Martín-Diana, Rico, & Barry-Ryan, 2008;
Zhan et al., 2012).
The experimental hypothesis was that by inhibition of enzy-matic
browning we will be able to preserve the consumer quality of mung bean
sprouts (Vigna radiata). The study consisted in char-acterization of the
inhibitory profile of PPO, selection of an optimal inhibitor and its
concentration, and assessment of the impact of post-harvest treatment on the
degree of enzymatic browning, phe-nolic content, and antioxidant capacity of
sprouts. However, ascor-bic acid is a well-known inhibitor of PPO from fruit
and vegetables; so far this is the first successful attempt aimed to use this
strategy in stored sprouts. The aim of the study was to select the optimal type
and concentration of polyphenol oxidase inhibitor and to evaluate its
application for inhibition of enzymatic browning dur-ing storage of mung
bean sprouts.
2. Material and methods
2.1. Chemicals
0
ABTS (2,2 -azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), a-
amylase, pancreatin, pepsin, bile extract, L-cysteine, citric acid, ascorbic acid,
catechol, sodium hypochlorite, trichloroacetic acid, dithiothreitol, and the Folin-
Ciocalteau reagent were purchased from Sigma–Aldrich company (Poznan,
Poland). All other chemi-cals were of analytical grade.
2.2. Materials
Mung bean seeds were purchased from the PNOS S.A. in Ozarów
Mazowiecki, Poland. The seeds were sterilized in 1% (v/v) sodium
hypochlorite for 10 min, dried, and washed with distilled water until they
reached neutral pH. Next, the seeds were placed in dis-tilled water and soaked
for 6 h at 25 LC. The seeds (approximately 150 per plate, 8 grams) were dark-
germinated in a growth cham-ber (SANYO MLR-350H) on Petri dishes (125
mm) lined with absor-bent paper (relative humidity 70%, 25 LC). Seedlings
were watered daily with 5 mL of Milli-Q water. After 3 days of germination,
sprouts were manually collected, rapidly frozen, lyophilized, ground in a
laboratory mill, sieved (60 mesh), and kept in poly-ethylene bags at 20 LC
(unless stated otherwise). For the posthar-vest storage experiment, 3-day-old
sprouts were manually collected and washed for 2 h in 0.2 mM, 2 mM, and 20
mM solu-tions of ascorbic acid. After that, the sprouts were kept in
polypropylene boxes at 4 LC for 7 days. After 3 and 7 days of stor-age, the
sprouts were collected from the boxes, rapidly frozen, lyo-philized, ground in
a laboratory mill, sieved (60 mesh), and kept in polyethylene bags at 20 LC.
2.3. Analysis of the browning index
The browning index was determined using the procedure described by
(Kim et al., 2014)). Briefly, 2 g of the sample was homogenized in 20 mL of
10% trichloroacetic acid, allowed to stand for 2 h at 35 LC, and centrifuged at
15,000 g at 25 LC for 15 min.
The absorbance of the supernatant was measured at 420 nm on a
spectrophotometer (UV-1280, Shimadzu).
2.4. Activity of polyphenol oxidase
2.4.1. Extract preparation
All extraction procedures were conducted at 4 LC. For polyphe-nol
oxidase (PPO), 2 g of the fresh sample was ground with 8 mL of 100 mM
sodium phosphate buffer (pH 5.8; optimal pH for PPO activity in sprouts
previously determined using sodium phosphate buffer) containing 0.2 g of
polyvinylpyrrolidone. The extracts were then homogenized and centrifuged at
12,000 g at 4 LC for 30 min. The supernatants were used for further analysis
´
(Gawlik-Dziki, Złotek, & S wieca, 2008).
2.4.2. Polyphenol oxidase (PPO) assay
For the PPO assay, 100 lL of the extract was incubated with 2 mL 0.05 M
phosphate buffer (pH 5.8) and 0.5 mL of 0.5 M catechol at 24 LC for 5 min,
and absorbance at 398 nm was measured. The PPO activity was expressed as
U, where U = 0.001 DOD398/min under the conditions of the assay
(Galeazzi, Sgarbieri, & Constantinides, 1981).
2.4.3. Impact of various inhibitors on PPO activity in mung bean sprouts
The inhibitory effects of ascorbic acid, citric acid, and L-cysteine on PPO
activity were examined. Three different concentrations of these inhibitors
(0.2, 2 and 20 mM) were tested using 0.05 M of catechol as a substrate. The
corresponding control contained the same concentration of the enzyme, in the
absence of the inhibitor. Percentage inhibition was calculated using the
following equation:
Inhibitionð%Þ ¼ ðA0 Ai=A0Þ 100%; where :
A0—initial PPO activity ðwithout the inhibitorÞ
Ai PPO activity with the inhibitor
2.5. Analysis of phenolics and antioxidant capacity
2.5.1. Extraction procedures
2.5.1.1. Chemical extraction. Lyophilized samples of mung bean sprouts (500
mg of dry mass) were subsequently extracted for 1 h at room temperature
(300 rpm) in a capped centrifuge tube with 5 mL of different solvents: 50%
methanol, 60 mM HCl in 50% methanol, and finally with 60 mM HCl in 70%
acetone. The mixture was centrifuged (15 min, 3000 g, 22 LC) and the super-
natants from all steps were combined.
2.5.1.2. Digestion in vitro. In vitro digestion was performed as described
previously (Minekus et al., 2014). For simulated mastica-tion and
gastrointestinal digestion, germinated sprouts (500 mg of lyophilized sprouts)
were homogenized in 0.5 mL PBS buffer and 1 mL of simulated salivary fluid
[15.1 mmol/L KCl, 3.7 mmol/L KH 2-PO4, 13.6 mmol/L NaHCO3, 0.15
mmol/L MgCl2 (H2O)6, 0.06 mmol/L (NH4)2CO3, 1.5 mmol/L CaCl2, a-
amylase (75 U/ mL)] and shaken for 10 min at 37 LC. Next, the samples were
adjusted to pH 3 with 6 M HCl, suspended in 2 mL of simulated gastric fluid
[6.9 mmol/L KCl, 0.9 mmol/L KH2PO4, 25 mmol/L NaHCO3, 47.2 mmol/L
NaCl, 0.1 mmol/L MgCl2 (H2O)6, 0.5 mol/L (NH4)2CO3 0.15 mmol/L CaCl2,
pepsin (2000 U/mL)] and shaken for 120 min. at 37 LC. After simulated
gastric digestion, the samples were adjusted to pH 7 with 1 M NaOH and
suspended in 4 mL simulated intestinal fluid [6.8 mmol/L KCl, 0.8 mmol/L
KH2PO4, 85 mmol/L
NaHCO3, 38.4 mmol/L NaCl, 0,33 mmol/L MgCl2 (H2O)6,
0.15 mmol/L CaCl2, 10 mol/L bile extract, pancreatin (2000
 1162 ´
M. Sikora, M. Swieca / Food Chemistry 239 (2018) 1160–1166
U/ mL)]. The prepared samples underwent in vitro intestinal diges-tion for
120 min.
2.5.2. Phenolic content (TPC)
The amount of phenolics was determined using the Folin-Ciocalteau
reagent (Singleton, Orthofer, & Lamuela-Raventós, 1999). The amount of
phenolics was expressed as gallic acid equiv-alents (GAE) in mg per g of dry
mass (d.m.).
2.5.3. Antiradical activity (ABTS)
The experiments were carried out using the ABTS decolorization assay
(Re et al., 1999). The free radical scavenging ability was expressed as Trolox
equivalents in mg per g of sprout dry mass (d.m.).
2.5.4. Reducing power (RP)
Reducing power was determined with the method of Pulido, Bravo, and
Saura-Calixto (2000). Reducing power was expressed as Trolox equivalents
in mg per g of sprouts dry mass (d.m.)
2.5.5. Theoretical approach
The following factors were determined for better understanding of the
relationships between biologically active compounds in the light of their
bioaccessibility (Gawlik-Dziki et al., 2015).
– the relative phenolic bioaccessibility factor (RBF):
RBF ¼ CD=CCE
where: CD – concentration of compounds after simulated gas-trointestinal
digestion, CCE – concentration of compounds after chemical extraction,
– the relative antioxidant efficiency factor (REF):
REF ¼ AD=ACE
where: AD – activity of the extract after simulated gastrointesti-nal
digestion, ACE – activity of the chemical extract.
2.6. Extraction and determination of ascorbic acid content
Ascorbic acid content was determined according to the methods described
earlier by Campos, Ribeiro, Della Lucia, Pinheiro-Sant’Ana, and Stringheta
(2009) with some modification (Złotek
´
& S wieca, 2016). Ascorbic acid content was expressed in mg per 1 g of dry
mass (DM).
2.7. Statistic analysis
All experimental results were mean ± S.D. of three independent
experiments (n = 9). One-way analysis of variance (ANOVA) and Tukey’s
post hoc test were used to compare groups (seeds as well as control and
elicited sprouts) (STATISTICA 6, StatSoft, Inc., Tulsa, USA). Differences
were considered significant at p < 0.05.
3. Result and discussion
Enzymatic browning is one of the most important reactions resulting in
negative changes in the color, taste, flavor, and nutri-tional value of fruits and
vegetables (Severini et al., 2003). The study was focused on improving
consumer quality of mung bean by inhibition of enzymatic browning.
3.1. Impact of various inhibitors on the PPO activity
The impact of various inhibitors of PPO activity (commonly used in food
industry such as citric acid, ascorbic acid, and
L-cysteine(Garcia & Barrett, 2002)) from mung bean sprouts is pre-sented in Table
1. The addition of ascorbic acid to the reaction mix resulted in a concentration-
dependent inhibition of PPO activity, although complete inactivation could not be
achieved. Ascorbic acid in the highest concentration (20mM) reduced the activity
of PPO to 2.61%. PPO was almost completely inactivated by 20 mM L-cysteine.
The weakest inhibitor of PPO from the mung sprouts was citric acid, which in the
highest concentration lowered polyphenol oxidase activity only to 47,18% (Table
1).
So far, L-cysteine has been reported to be an effective inhibitor of PPO
obtained from different fruits and vegetables. Citric acid effectively inhibited
the native PPO from lettuce (Altunkaya & Gökmen, 2009) and apple
(Pizzocaro, Torreggiani, & Gilardi, 1993). Our results are in agreement with
those published by Holzwarth et al. (2013), who reported that the activity of
PPO from strawberries (Fragaria x ananassa Duch.) was completely inhibited
by 0.08% L-cysteine. L-cysteine was also the most effective inhibi-tor. The
effect of sodium metabisulphite, citric acid, sodium bisul-phate, and ascorbic
acid (0,01–0,3 mol/L) on inhibition of oxidative browning in sweet potato
was studied by Arogundade and Mu (2012). They proved that the highest
inhibition of PPO was observed using citric and ascorbic acid.
3.2. Effects of ascorbic acid treatment on the browning index and PPO
activity in stored mung bean sprouts
Based on the results of the inhibitory profile of PPO for post-harvest
treatment, ascorbic acid was selected. This compound, besides its high
inhibitory capacity against PPO, is known as an effective antioxidant and
antimicrobial agent (Duarte & Lunec, 2005). It has been shown (Fig. 1) that
the activity of PPO increased during storage. Postharvest treatment of sprouts
with ascorbic acid significantly inhibited the activity of the enzyme after 3
days of storage. Sprouts washed with 2 mM and 20 mM ascorbic acid were
characterized by lowered activity of PPO, i.e. reduced by 51% and 60%,
respectively, compared to the control. These results are in agreement with
those published by Arogundade and Mu (2012), who reported that an increase
in the concentration of an oxidative browning inhibitor decreases the
browning index in sweet potato. Most importantly, the inhi-bition was
reflected in a significant decrease in enzymatic browning (Fig. 1). Similar
results were obtained by Ali, Khan, and Malik (2016), who observed that the
browning index of Litchi fruits (Litchi chinensis Sonn) continuously
increased throughout the storage period from 7 days to 28 days and was
positively correlated with PPO activities. Unfortunately, after 7 days of
storage there were no differences between the control and treated sprouts.
Table 1
Impact of various inhibitors on the PPO activity in mung bean sprouts.
Inhibitor concentration [mM] Relative activity [%]
Control – 100.00 ± 0.85g
Citric acid 0.2 93.24 ± 6.48f
2 77.76 ± 9.00e
20 47.18 ± 5.29d
Ascorbic acid 0.2 16.60 ± 2.12c
2 8.76 ± 0.85bc
20 2.61 ± 1.16a
L-cysteine 0.2 9.51 ± 0.79c
2 6.15 ± 2.37b
20 0.91 ± 0.30a
Means (±SD) with different letter are significantly different (n = 9; a = 0.05).
 ´ 1163
M. Sikora, M. Swieca / Food Chemistry 239 (2018) 1160–1166
PPO activity Browning index

800 de 0.7
700 cd
cd 0.6
CD CD C
600 C bc C C
C 0.5

index
c be

[Abs 420nm]
[U/mg] PPO activity 500 ab AB A 0.4

Browning
300
ab a
400
0.3
200 0.2

100 0.1

0 0.0

3F 3S 3S0.2 3S2 3S20 7S 7S0.2 7S2 7S20

Fig. 1. Effects of ascorbic acid post-harvest treatment on the browning index and PPO activity in stored mung bean sprouts Means with different letters (uppercase - browning index and lowercase –
PPO activity) are significantly different (n = 9; a = 0.05). 3 F- 3-day-old fresh sprouts, 3S-3-day-old sprouts stored for 3 days, 3S0.2, 3S2, 3S20- 3-day-old sprouts washed in 0.2 mM, 2 mM, and 20
mM ascorbic acid, respectively, and stored for 3 days. 7S- 3-day-old sprouts stored for 7 days, 7S0.2, 37S2, 7S20- 3-day-old sprouts washed in 0.2 mM, 2 mM, and 20 mM ascorbic acid, respectively,
and stored seven days. PPO-polyphenol oxidase, Abs-absorbance.

3.3. Effect of PPO inhibition on phenolics and ascorbic acid content in mung
bean sprouts
PPO oxidases phenolic compounds, thus it may influence on the
antioxidant capacity of food. In the study, the influence of treat-ments and
storage on phenolic content and antioxidant capacity was determined for a
chemically extractable and potentially bioac-cessible fraction. An increase in
chemically extractable phenolics was only observed in the stored sprouts
treated with 20 mM ascor-bic acid; compared to the control, there was an
increase by ca. 22% and 23% after 3 and 7 days of storage, respectively.
Taking into account the potentially bioaccessible fraction, an increase was
only found after 3 days of storage of the sprouts treated with 2 and 20 mM
ascorbic acid. Most importantly, storage and ascorbic acid treatment exerted a
positive effect on the bioaccessibility of TPC (Table 2). To exclude the
interference of exogenous ascorbic acid in the Folin-Ciocalteu assay, its
content was determined in the samples as well. According to the results, it is
clearly visible that the increase in the ascorbic acid content was caused by de
novo synthesis. There was also no relationship between the determined
contents of chemically extractable phenolics and ascorbic acid. It may be
suggested that the increase in the phenolic content in the sprouts treated with
20 mM ascorbic acid was caused by oxida-
tive stress. There are some reports that support this suggestion that
acidification can induce a natural mechanism of resistance e.g. synthesis of
low molecular weight antioxidants (Burguieres, McCue, Kwon, & Shetty,
2007).
Germination resulted in a decrease in the chemically extracta-ble phenolic
compounds by ca. 20%. These results are incompatible with those published
by Paja˛ k et al. (2014) who reported that total phenolic content increased
during germination. These differences may result from the diversity among
the varieties, growing and storage conditions, as well as the extraction
procedures. On the other hand, in the first step of sprouting (inhibition), the
phenolics level was decreased due to leaching of solid matter in soaking water
(Ghavidel & Prakash, 2007).
3.4. Effect of PPO inhibition on antioxidant capacity of mung bean sprouts
An increase in low-molecular weight antioxidants is usually reflected in
elevation of antioxidant capacity. The results concern-ing the effect of the
treatments used and storage are presented in Figs. 2 and 3. Compared to the
dry seeds, the fresh sprouts had a significantly lower reducing power. The
reducing capacity of the sprouts did not increase during the storage time but
was affected

Table 2
Effect of PPO inhibition on phenolics and ascorbic acid contents in fresh and stored mung sprouts.

Chemically extractable phenolics Potentially bioaccessible phenolics Relative phenolics bioaccessibility Ascorbic Acid
[mg/g d.m.] [mg/g d.m.] factor [mg/g d.m.]
Seeds 9.03 ± 0.27d 6.58 ± 0.24b 0.73 8.44 ± 0.76a
3F 7.35 ± 0.32ab 6.07 ± 0.10a 0.83 135.62 ± 22.65c
3S 7.34 ± 0.23ab 6.09±.010a 0.83 128.14 ± 18.12c
3S0.2 7.43 ± 0.28ab 6.10 ± 0.06a 0.82 148.43 ± 19.63cd
3S2 7.10 ± 0.23a 8.71 ± 0.07cd 1.23 81.16 ± 6.04b
3S20 8.94 ± 0.52d 10.39 ± 0.31de 1.16 77.95 ± 13.59b
7S 7.68 ± 0.22abc 9.17 ± 0.17cd 1.19 84.36 ± 7.55b
7S0.2 7.69 ± 0.27abc 8.41 ± 0.11c 1.09 92.90 ± 13.59bc
7S2 7.88 ± 0.14abc 9.01 ± 0.58cd 1.14 75.82 ± 10.57b
7S20 9.46 ± 0.14de 9.70 ± 0.85de 1.02 109.99 ± 4.53bc

Means with different letter within the same columns are significantly different (n = 9; a = 0.05).
3 F- 3-day-old fresh sprouts, 3S- 3-day-old sprouts stored for 3 days, 3S0.2, 3S2, 3S20- 3-day-old sprouts washed in 0.2 mM, 2 mM, and 20 mM ascorbic acid, respectively, and stored for 3 days.

7S- 3-day-old sprouts stored for 7 days, 7S0.2, 37S2, 7S20- 3-day-old sprouts washed in 0.2 mM, 2 mM, and 20 mM ascorbic acid, respectively, and stored seven days.
PPO- polyphenols oxidase; d.m. – dry mass.
1164 ´
M. Sikora, M. Swieca / Food Chemistry 239 (2018) 1160–1166
16 Chemically extractable fraction

14 Potentially bioaccessible fraction C

C
12 B

10 A A

A A A A A
Reducing power
[mg TE/1g d.m.]
8

d d
6
c c b

4 b bc
a ab a

2
0

Seeds 3F 3S 3S0.2 3S2 3S20 7S 7S0.2 7S2 7S20

Fig. 2. Effects of ascorbic acid post-harvest treatment on the reducing power of stored mung bean sprouts Means with different letter are significantly different (n = 9; a = 0.05). 3 F- 3-day-old fresh
sprouts, 3S- 3-day-old sprouts stored 3 days, 3S0.2, 3S2, 3S20- 3-day-old sprouts washed in 0.2 mM, 2 mM and 20 mM ascorbic acid and stored 3 days, respectively. 7 S- 3-day-old sprouts stored seven days, 7S0.2,
37S2, 7S20- 3-day-old sprouts washed in 0.2 mM, 2 mM and 20 mM ascorbic acid and stored seven days, respectively. TE-Trolox equivalents, d.m.-dry mass.

8
D Chemically extractable fraction
7 Potentially bioaccessible fraction
6 C

C
BC C BC
5 AB ABC
AB
Antiradical activity

4 A

ab ab
[mgTE/1gd.m.]

3 b b ab a b

a a a
2
1

Seeds 3F 3S 3S0.2 3S2 3S20 7S 7S0.2 7S2 7S20

Fig. 3. Effects of ascorbic acid post-harvest treatment on the antiradical activity in stored mung bean sprouts. Means with different letters are significantly different (n = 9; a = 0.05). 3F- 3-day-old fresh
sprouts, 3S- 3-day-old sprouts stored for 3 days, 3S0.2, 3S2, 3S20- 3-day-old sprouts washed in 0.2 mM, 2 mM, and 20 mM ascorbic acid, respectively, and stored for 3 days. 7S- 3-day-old sprouts stored for 7 days,
7S0.2, 37S2, 7S20- 3-day-old sprouts washed in 0.2 mM, 2 mM, and 20 mM ascorbic acid, respectively, and stored seven days. TE-Trolox equivalents, d.m.-dry mass.

by the ascorbic acid treatment. The highest values were noted on the 3rd and
7th days of storage in the sprouts treated with 20 mM ascorbic acid (Fig. 2).
Sprouting caused a decrease in the anti-radical properties (both chemically
extractable and potentially bioaccessible fraction) compared to the dormant
seeds. There was no effect of ascorbic acid treatment on the antiradical
capacity of stored sprouts. Compared to the respective controls, the antirad-
ical capacity of the chemically extractable fraction of the sprouts treated with
20 mM ascorbic acid increased statistically during storage. After 3 and 7 days,
an increase by 16.2% and 7.7% was determined, respectively (Fig. 3).
According to Table 3, it is clearly visible that the antioxidant capacity was
weakly bioaccessible in vitro. The storage significantly affected the
bioaccessibility of the compound exhibiting antiradical properties (RBF value
0.71, 0.47, and 0.47 for the fresh and 3- and 7-day stored control sprouts,
respectively). Compared to the respective control, an increase in the
bioaccessibility of reducing potential was found after 3 days of storage;
however, on the 7th day no such a clear effect was observed.
It may be suggested that ascorbic acid, being an excellent antioxidant in
the food system, helps to maintain the active state for many bioactive
compounds, such as vitamin E, flavonoids, and some phenolics (Burguieres et
al., 2007). The increase in the reducing power in the sprouts treated with 20
mM ascorbic acid is rather linked with the elevated level of phenolics than the
direct action of endogenous ascorbic acid. The results of the antioxidant
capacity of mung bean sprouts are in agreement with those pub-lished by S
´
wieca and Gawlik-Dziki (2015), who reported that three-day
sprouting reduces antiradical activity compared to that of
dormant seeds. The antioxidant capacity of two lettuce
cultivars
 ´ 1165
M. Sikora, M. Swieca / Food Chemistry 239 (2018) 1160–1166

Table 3 Altunkaya, A. (2011). Effect of whey protein concentrate on phenolic profile and browning of
Influence of storage and treatment on potential bioaccessibility of antioxidant capacity in mung fresh-cut lettuce (Lactuca sativa). Food Chemistry, 128(3), 754–760.
bean sprouts. Altunkaya, A., & Gökmen, V. (2009). Effect of various anti-browning agents on phenolic
compounds profile of fresh lettuce (L. sativa). Food Chemistry, 117(1), 122–126.
The relative antioxidant efficiency factor (REF)
Antiradical activity Reducing power Arogundade, L. A., & Mu, T. H. (2012). Influence of oxidative browning inhibitors and isolation
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Seeds 0.37 0.20 1374–1384.
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Peñas, E., Gómez, R., Frías, J., & Vidal-Valverde, C. (2010). Effects of combined treatments of
high pressure, temperature and antimicrobial products on germination of mung bean seeds
Conflict of interest statement and microbial quality of sprouts. Food Control, 21(1), 82–88.

The authors declare no conflict of interest.
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