LWT - Food Science and Technology 117 (2020) 108614

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LWT - Food Science and Technology

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Optimization of extraction process of antioxidant compounds from yellow T

onion skin and their use in functional bread production
Tomasz Piechowiaka,∗, Katarzyna Grzelak-Błaszczykb, Radosław Bonikowskic, Maciej Balawejdera
a
Department of Chemistry and Food Toxicology, Faculty of Biology and Agriculture, University of Rzeszow, St. Cwiklinskiej 1a, 35-601, Rzeszow, Poland
b
Institute of Food Technology and Analysis, Faculty of Biotechnology and Food Sciences, Lodz University of Technology, St. Stefanowskiego 4/10, 90-924, Łodz, Poland
c
Institute of General Food Chemistry, Faculty of Biotechnology and Food Sciences, Lodz University of Technology, St. Stefanowskiego 4/10, 90 – 924, Łodz, Poland

A R T I C LE I N FO A B S T R A C T

Keywords: The major aim of this research was to demonstrate a possible use of antioxidant compounds isolated from onion
Antioxidants skin in functional bread production. The scope of the research included a study on the extraction process of
Bread antioxidant compounds from onion skin and determination of optimal process conditions using response surface
Extraction methodology, as well as the investigation of antioxidant properties of bread enriched with onion skin extract.
Flavonoids
Research showed that total yield of extraction of antioxidant compounds from onion skin using methanol as a
Onion
solvent was dependent on the applied process conditions i.e. temperature and process time, as well as the ratio of
onion skin mass to methanol volume. From 1 g sample of onion skin, the highest yield is achieved at 145 min of
process time under 44 °C, using 30 mL of methanol. The obtained onion skin extract was characterized by high
antioxidant activity (441.4–593.9 mg QE g−1) which was shaped by high concentration of flavonoid compounds
i.e. quercetin (315.6 mg g−1), quercetin-3-glucoside (40.3 mg g−1), isoramnethin (14.8 mg g−1) and kaempferol
(10.9 mg g−1). Furthermore, it was found that antioxidant activity of wheat bread enriched with onion skin
extract was closely correlated with extract dose.

1. Introduction transition metals (Piechowiak & Balawejder, 2019). Furthermore,

Growth of nutritional awareness in consumers causes that they draw
more and more attention to nutritious and health-promoting values of
food. An increasing interest in food, which also contains a high anti-
oxidant potential apart from providing basic nutrients is currently ob-
served. According to scientific reports, antioxidant deficiency in a daily
diet is a cause for organism ageing and occurrence of numerous diseases
of affluence being a consequence of oxidative stress. Therefore, it is
justifiable to search for antioxidant sources which would efficiently
neutralize a surplus of free radicals after consumption (Piechowiak &
Balawejder, 2019).
The onion skin is a waste formed during onion production. The
feature of such material, as it turns out, is high content of compounds
with an antioxidant properties, primarily polyphenols. The level of such
compounds was determined to be significantly higher than a specific
part of the plant and it amounted to 2–10 g/kg of the skin weight and it
depended upon its variety, climate, soil and agrotechnical conditions of
growing. A large number of in vitro studies have suggested that poly-
penols are reported to possess strong free radical scavenging activity
based on their ability to act as hydrogen or electron donors and chelate
quercetin isolated from onion skin showed an anti-inflammatory, an-
timicrobial and anticancer properties (in vitro and in vivo) (Świeca,
Gawlik-Dziki, Dziki, Baraniak, & Czyż, 2013). It can therefore be stated
that composition of the bioactive compounds isolated by means of an
appropriate extraction method will constitute a reliable source of
agents positively influencing a human organism (Benitez et al., 2011;
Gawlik-Dziki et al. 2014, 2015; Viera et al., 2017).
Conventional solvent extraction is commonly used in many bran-
ches of food and pharmaceutical industry. However, the efficiency of
extraction of antioxidant compounds from plant materials depends on
the process conditions used, such as: temperature and process time, as
well as type and concentration of the solvent in extraction mixture.
According to the current state of knowledge, the phenolic compounds
contained in onion skin are characterized by low water solubility. In the
research work to date, flavonoids (quercetin) have been isolated from
onion skin by conventional solvent extraction, with the use of mainly
aqueous solutions of ethanol (Jin et al., 2011, Zubairu & Mshelia,
2015). It should also be mentioned that the removal of water from a
product is highly energy-intensive and the cost of ethanol per unit is
relatively high, too. Methanol has a lower boiling point than ethanol

∗
Corresponding author.
E-mail address: tomek.piechowiak92@gmail.com (T. Piechowiak).

https://doi.org/10.1016/j.lwt.2019.108614
Received 30 June 2019; Received in revised form 8 August 2019; Accepted 10 September 2019
Available online 11 September 2019
0023-6438/ © 2019 Elsevier Ltd. All rights reserved.
T. Piechowiak, et al. LWT - Food Science and Technology 117 (2020) 108614

and can be easily vaporized after extraction. Therefore, the use of extract (Y3) was examined (Table 1), (Balawejder & Piechowiak, 2018,
methanol to extraction of antioxidants from onion skin is fully justified. p. 424683; Pandey, Belwal, Sekar, Bhatt, & Rawal, 2018).
The daily consumption of bread and bakery goods is estimated to The extraction process was conducted in batch extractor, equipped
make up ca. 70% of the total cereal consumption. Cereal products are with mechanical stirrer (speed: 300 rpm), and temperature sensor. The
good source of carbohydrate, protein, unsaturated fatty acids and obtained crude-extracts were filtered in vacuum filtration apparatus,
dietary fiber, but they are poor in antioxidants. Many bioactive com- evaporated and dried in rotary evaporator (45 °C, 300 mbar, 90 rpm).
pounds found in the grain, especially polyphenols, are particularly The experiment was performed in triplicate.
concentrated in the bran layer, which is discarded during the produc- Experimental data were tested using multiple regression and ana-
tion of bread and cake flour. In addition, the most of cereal antioxidants lysis of variance (ANOVA), the effects of which allowed to determine
are sensitive to heat treatment. For this reason, bakers and scientists are significance of the influence of extraction process conditions (Xi) on the
involved in optimizing a bread making technology to improve the dependent variables (Yi) and generation of appropriate mathematical
biological value of final products. Among the ingredients that could be models (α = 0.05). The generalized linear (1) and second-order poly-
included in bread formulation are herbs, species, dried fruits and ve- nomial models (2) were used in the RSM:
getables as well as plant extracts with high antioxidant activity (Peng 1
et al., 2010). Y = β0 + ∑ βi Xi
The major aim of this research was to demonstrate a possible use of i=1 (1)
antioxidant compounds extracted from onion skin in functional bread
3 3 3
production. The scope of the research included a study on the extrac-
tion process of antioxidant compounds from onion skin using methanol
Y = β0 + ∑ βi Xi + ∑ βii Xi2 + ∑ βij Xi Xj
i=1 i=1 i<j=1 (2)
as an extraction solvent and determination of optimal process condi-
tions, as well as the investigation of antioxidant properties of wheat where, Y is the dependent parameter, β0, βi βii – regression coefficient,
bread enriched with onion skin extract. XiXj – independent non-coded variables.
The research results were analysed using Design-Expert 10.0.4.
software (Statease, Minneapolis, MN, USA).
2. Materials and methods

2.1. Optimization of extraction process of bioactive compounds from onion
skin
2.1.1. Plant material and experimental design
Research material was yellow onion skin (Allium cepa L.) purchased
from local onion-producer. The onion skin was characterized by neutral
smell and taste, without any signs of mold infestation. Before extrac-
tion, onion skin was dried in convection dryer (55 °C, 120 min, 2 m/s)
and ground to an average particle size of 2 mm.
Optimization of extraction of antioxidant compounds from onion
skin was performed by means of a Response Surface Methodology
(RSM), using three-factor, three-level compositional plan. An influence
of extraction temperature (X1), time of process (X2) and ratio of onion
skin mass to methanol volume (X3) on the total yield of extraction (Y1),
phenolic compounds recovery (Y2) and antioxidant activity of dried
2.1.2. Analysis of the extract
2.1.2.1. Total yield of extraction. Total yield of extraction [g·100 g−1]
was calculated as follows:
Wextract
YE = ⋅100, where:
Wsample (3)
Wextract and Wsample are the weight of onion skin extract [g] and
onion skin [g], respectively.
2.1.2.2. Total polyphenols content. To 50 μL of onion skin extract
methanol solution (50 mg 100 mL−1), 950 μL of distilled water, 50 μL
of Folin-Ciocalteu reagent (Chempur, Poland) diluted with distilled
water (1:1 v/v) and 100 μL of 7% Na2CO3 (Chempur, Poland) solution
were added. The reaction mixture was incubated in darkness for 30 min
and then the absorbance was measured at 750 nm. The obtained results

Table 1
Optimization experience plan using RSM and the research results as mean values ( x‾ ± SD, n = 3).
No Temp.[°C] Time [min.] Ratio of onion skin mass to methanol Total yield of extraction [g Polyphenols recovery [g Antioxidant activity DPPH• [mg
volume [g ml−1] 100g-1] 100 g−1] g−1]

X1 X2 X3 Y1 Y2 Y3

1 22 (−1) 180 (1) 0.055 (1) 3.86 ± 0.01 2.44 ± 0.01 199.84 ± 12.42
2 50 (1) 180 (1) 0.055 (1) 9.87 ± 0.22 3.02 ± 0.83 220.40 ± 11.12
3 36 (0) 115 (0) 0.044 (0) 5.24 ± 0.44 3.18 ± 0.11 434.68 ± 21.42
4 59.5 (1.68) 115 (0) 0.044 (0) 8.45 ± 0.51 3.02 ± 0.84 308.78 ± 22.54
5 36 (0) 115 (0) 0.044 (0) 6.08 ± 0.26 2.89 ± 0.01 390.96 ± 11.42
6 22 (−1) 50 (−1) 0.033 (−1) 3.96 ± 0.11 2.32 ± 0.12 306.80 ± 45.12
7 36 (0) 115 (0) 0.026 (−1.68) 9.44 ± 0.83 3.43 ± 0.54 452.89 ± 35.54
8 22 (−1) 50 (−1) 0.055 (1) 3.69 ± 0.35 2.69 ± 0.33 118.47 ± 10.43
9 36 (0) 224 (1.68) 0.044 (0) 8.62 ± 0.76 2.96 ± 0.12 271.86 ± 28.76
10 36 (0) 115 (0) 0.044 (0) 6.31 ± 0.16 3.08 ± 0.89 442.43 ± 11.53
11 22 (−1) 180 (1) 0.033 (−1) 3.94 ± 0.32 2.53 ± 0.11 224.87 ± 26.31
12 36 (0) 5.7 (−1.68) 0.044 (0) 5.52 ± 0.33 2.30 ± 0.04 353.97 ± 13.32
13 36 (0) 115 (0) 0.044 (0) 6.86 ± 0.96 3.02 ± 0.03 401.91 ± 4.12
14 12.5 (−1.68) 115 (0) 0.044 (0) 2.64 ± 0.32 1.64 ± 0.12 188.16 ± 11.34
15 50 (1) 180 (1) 0.033 (−1) 10.43 ± 0.1 3.22 ± 0.55 336.19 ± 13.54
16 50 (1) 50 (−1) 0.033 (−10 8.71 ± 1.11 2.66 ± 0.12 365.03 ± 18.43
17 50 (1) 50 (−1) 0.055 (1) 7.94 ± 0.98 2.03 ± 0.34 255.87 ± 12.54
18 36 (0) 115 (0) 0.062 (1.68) 4.63 ± 0.12 2.36 ± 0.22 268.22 ± 33.36
Adjusted determination coefficient - Adj-R2 0.81* 0.79* 0.81*
Lack of fit test results (p-value), α = 0.05 0.21 0.054 0.152

*-significant, p < 0,05.

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T. Piechowiak, et al. LWT - Food Science and Technology 117 (2020) 108614

were expressed as quercetin equivalent (QE, Sigma-Aldrich, Darmstadt, Latimer, 2007).

Germany), (Ozyurt, Demirata, & Apak, 2011).
2.1.2.3. Total flavonoids content. To 400 μL of the extract, 40 μL of 5%
NaNO2 (Chempur), 40 μL of 10% AlCl3 (Chempur), 400 μL of 4% NaOH
(Chempur) and 120 μL of distilled water were added. The reaction
mixture was incubated in darkness for 15 min and next the absorbance
was measured at 430 nm. The flavonoids content was determined based
on the calibration curve for the quercetin solutions in the range of
10–500 μM (Benitez et al., 2011).
2.1.2.4. Antioxidant activity against DPPH• radicals. To 1 mL of 100 μM
DPPH• radical solution (Sigma-Aldrich), 30 μL of onion skin extract
methanol solution (5 mg 100 mL−1) was added. After 30 min of
incubation in darkness, the absorbance was measured at 515 nm.
Quercetin (0–250 μM concentration) was used for calibration and the
results were expressed as mg of quercetin equivalent (QE) per 1 g of
dried onion skin extract (Ak & Gulcin, 2008).
2.1.2.5. Antioxidant activity against ABTS•+ radicals. A 7 mM of ABTS
(Sigma-Aldrich) in 2.45 mM of K2S2O8 water solution was prepared.
Next, the radical solution was incubated in darkness for 24 h. Before the
analysis, ABTS•+ solution was diluted with distilled water until the
absorbance 0.7 ± 0.02, at λ = 400 nm.
To 1 mL of ABTS•+ solution, 10 μL of the extract was added. After
15 min of incubation in darkness, the absorbance of the solution was
measured at λ = 400 nm. Quercetin (0–500 μM concentration) was
used for calibration and the results were expressed as mg of quercetin
equivalent (QE) per 1 g of onion skin extract (Ak & Gulcin, 2008).
2.2. Preparation and analysis of bread enriched with onion skin extract
2.2.1. Bread making
For bread making, 400 g of wheat bread flour (0.75% ash content,
14% humidity) mixed with 120 mL of water, 40 mL of 10% NaCl, 40 mL
of water suspension containing 12 g of baker's yeast and dried onion
skin extract at the concentration of 0 (control sample), 0.1, 0.25, 0.5%
(in relation to the flour weight) for 6 min using a planetary mixer. After
fermentation (35 °C, 1 h), the pieces of dough (100 g) were baked at the
temperature of 180 °C for 30 min. After baking, the bread samples were
cooled to room temperature for further analysis.
2.2.2. Determination of antioxidant activity of bread sample
Sample of 5 g of bread was homogenized in 50 mL of methanol (30 s,
20 000 rpm). The homogenate was shaken for 30 min (180 rpm) and
clarified by centrifugation at 10 000 g for 15 min. The obtained super-
natant was used to determine the total polyphenols content and anti-
oxidant activity using DPPH and CUPRAC method (see page 7–8).
The antioxidant activity of bread sample was expressed as trolox
equivalent (TE, Sigma-Aldrich) while, the polyphenols content as gallic
acid equivalent (GAE, Sigma-Aldrich) per 100 g of the bread.
2.2.3. Statistical analysis
Significance of differences was conducted using one way ANOVA
and the Tukey test (α = 0.05) with STATISTICA 13.0 software.

3. Results and discussion

2.1.2.6. Determination of the antioxidant activity using CUPRAC
method. To 0.25 mL of 1 mM CuCl2 (Sigma-Aldrich), 0.25 mL of
3.1. Optimization of extraction of antioxidant compounds from onion skin
7.5 mM neocuproine (Sigma-Aldrich), 0.25 mL of 1 M NH4Ac
(Chempur), 50 μL of the research sample and 0.20 mL of distilled
As part of the work, we tested the impact of three variables that are
water were added. The reaction mixture was incubated for 30 min at
essential to ensure efficiency, extract quality and extraction cost-ef-
room temperature in darkness. Next, the absorbance was measured at
fectiveness. The variables were temperature, time of process and the
λ = 450 nm (using a blank sample as a reference). Quercetin
solvent volume used. We determined that the result of the extraction
(0–500 μM concentration) was used for calibration and the results
process is made up of the amount of the dry substance derived from
were expressed as mg of quercetin equivalent per 1 g of the sample
100 g of onion skin, the recovery of phenolic compounds and the an-
(Ozyurt et al., 2011).
tioxidant activity of a unit of the extract mass against DPPH• radicals.
Table 1 shows the optimization experience plan and the test results
2.1.2.7. Flavonoids analysis using high-performance liquid chromatography
as mean values ( x‾ ± SD, n = 3). In view of the high result variability
(HPLC). HPLC system with UV–Vis/DAD detector (Thermo Scientific)
depending on the process conditions, we were making conjectures
was used to analyze the flavonoids in onion skin extract. Before injected
about a significant impact of the process parameters on the efficiency of
to HPLC the extract was diluted with methanol and filtered through a
the onion skin antioxidants. We carried out a certain statistical analysis
0.2 μm nylon filter. The content of flavonoids was determined by the
to verify this hypothesis.
area of the peak at 375 nm, measured from the UV–Vis/DAD detector
Based on the results of a multiple regression analysis, we selected
interfacing XTerra Shield RP18 column (5 μm, 4.6 mm × 250 mm).
the most suitable mathematical models for the experimental data ob-
Chromatography was performed at 1 mL min−1 of flow rate using 0.1%
tained. We used a linear model to describe the extraction yield results
formic acid, 60% methanol and water as the solvent. Sample peaks
and a model in the form of a second-order polynomial (p < 0.05) for
were quantified with the external standards method (1.2–20 μg mL−1
the recovery of polyphenols and the antioxidant activity in relation to
concentration). The determination coefficients of all calibration curves
the dry matter of the extract. We found out that the selected models
were R2 > 0.9984 (Jin et al., 2011).
were characterized by good matching confirmed by the high value of
The limits of detection (LOD) and limits of quantification (LOQ)
the adjusted coefficient of determination Adj.-R2 (Table 1). In addition,
were determined at a signal-to-noise ratio of 3:1 and 10:1, respectively,
in the analysis of the results of the test for a lack of fit, we found that for
for quercetin (LOD = 1.12 μg mL−1, LOQ = 3.44 μg mL−1), iso-
the adopted ranges of determining variables, the selected mathematical
ramnethin (LOD = 0.64 μg mL−1, LOQ = 1.95 μg mL−1), kaempferol
models can be used to predict the efficiency of extraction of bioactive
(LOD = 1.2 μg mL−1, LOQ = 4.98 μg mL−1) and quercetin-3-glucoside
compounds from onion skin (p > 0.05). The regression model equa-
(LOD = 0.69 μg mL−1, LOQ = 2.14 μg mL−1). The quercetin, kaemp-
tions (4)–(6) for actual values (after removing insignificant factors) take
ferol, isoramnethin and quercetin-3-glucoside standards were pur-
the following form:
chased from Sigma-Aldrich.
Y1 = 2.26 + 1.6⋅10−1X1 + 10−2X2 − 64.99X3 (4)
2.1.2.8. Basic chemical composition. The basic composition of the
extract was determined by AOAC methodology using the following Y2 = −6.4⋅10−2 + 1.26⋅10−1X1 + 10−3X2 + 54.54X3 + 2.3
procedures: protein content – 920.152; crude fat – 930.09; dry matter
⋅10−4X1 X2 − 10−3X12 (5)
and ash content – 940.26, total dietary fiber (TDF) – 985.29 (Horwitz &

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T. Piechowiak, et al. LWT - Food Science and Technology 117 (2020) 108614

Fig. 1. Impact of process conditions on the yield of extraction. Fig. 2. Impact of extraction process conditions on the phenolic compounds
recovery from onion skin.

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T. Piechowiak, et al. LWT - Food Science and Technology 117 (2020) 108614

Y3 = −4.43⋅102 + 29.2X1 + 14.27⋅103X3 − 3.57⋅10−1X12 − 1.1

⋅10−2X22 − 2.51⋅10−5X32 (6)
For the yield of extraction, we only noted linear effects between the
factors tested and the dependant variable. This means that the tem-
perature rise, the amount of the solvent used and the extension of the
process for the adopted range of variables increase the dry matter of the
extract. Moreover, when comparing coefficients across linear regression
models, we determined that among the factors analysed, the process
temperature exerted the greatest impact on the final results, whereas
the process duration was affected least (Fig. 1).
The recovery of phenolic compounds from 100 g of onion skins was
among 1.64 g–3.45 g, expressed as an equivalent of quercetin.
According to the regression equation obtained, the relationships be-
tween the extraction time, the ratio of the skin weight to the methanol
volume and the extraction of polyphenols were linear in character. For
temperature, however, we noted both significant linear and square ef-
fects and, as a result, the extraction of polyphenols had an increasing
trend, reaching 44 °C, and then did not vary significantly (Fig. 2).
Regression model analysis for the antioxidant properties of a dry
extract revealed that the antioxidant activity of the extract against
DPPH radicals increased with the extension of the extraction time, the
increasing share of methanol in the extraction mixture and the tem-
perature reaching a certain critical value. Then the activity gradually
decreased, probably due to the increasing concentration of the sub-
stances in the extract, i.e. carbohydrates, proteins, dietary fibre frac-
tions and minerals which do not exhibit antioxidant properties and/or
to the thermal degradation of the antioxidants (Fig. 3). This was con-
firmed by significant square effects between the factors analysed and
the dependant variable.
On the basis of response surface plots and of regression model
analysis, we determined the optimum extraction conditions under
which the extraction of polyphenols and the antioxidant activity of the
extract demonstrate the maximum value. For the extraction mixture, we
adopted a temperature equal to 44 °C, a ratio of the methanol volume to
the skin weight of 30:1 and the process time of 145 min.
In order to finally verify the correctness of the regression model
adopted and the optimum process parameters determined on the basis
of the model, we carried out three additional experiments. Based on the
regression model in question, we determined the presumed theoretical
value for the dry matter efficiency, the extraction of polyphenols from
100 g of onion skins and the antioxidant activity of the extract under
optimum process conditions. Next, we compared the results with the
values obtained from experiments (Table 2). We found out that the
results obtained are within the limits set by the relevant confidence
intervals. This confirms the correctness of the regression models
adopted and the selection of optimum process parameters.
In recent years, a number of research reports have been published
on the impact of different extraction techniques on the recovery of
bioactive compounds from onion skin as well as on the polyphenols
profile and the antioxidant activity of the extract (Jin et al., 2011; Ko,
Cheigh, Cho, & Chung, 2011; Viera et al., 2017). However, the impact
of batch extraction with the use of methanol on the extraction of an-
tioxidants from onion skin has not been thoroughly tested to date. Jin
et al. (2011) have researched into optimization of periodic extraction of
quercetin from onion skin with the use of aqueous solutions of ethanol
as well as with additional support of ultrasounds and microwaves. The
authors have found that among the different extraction techniques they
tested, microwave extraction allowed them to recover the largest
amount of quercetin per unit of onion skin weight in the shortest space
of time of extraction. However, the efficiency values determined by
them were explicitly lower than those obtained by us and other authors.
In studies involving recovery of quercetin from onion skin, Campone
et al. (2018) and Ko et al. (2011) in turn reported that they achieved a Fig. 3. Impact of process conditions on the antioxidant activity of onion skin
satisfactorily high recovery yield of quercetin by extraction with carbon extract.
dioxide and supercritical water. Although supercritical water extraction

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Table 2
Experimental data of the validation of predicted values at optimal process conditions.
Response Predicted value Confidence interval Experimental value
Yi
95% min. 95% max.

−1
Y1 - total yield of extraction [g dried extract·100g onion skin] 8.84 ± 1.04 7.91 9.76 8.23 ± 0.52
Y2 – phenolic compounds recovery [g quercetin·100 g−1onion skin] 3.43 ± 0.28 3.06 3.78 3.24 ± 0.25
Y3 – antioxidant activity [mg quercetin·g−1 extract] 414.91 ± 41.15 363.62 466.18 401.52 ± 23.02

Table 3 3.3. Antioxidant activity of wheat bread enriched with onion skin extract
Chemical composition and antioxidant activity of onion skin ex-
tract. The daily consumption of bread is estimated to make up ca. 70% of
Parameters x‾ ± SD, n = 3 the total cereal consumption. This means that bread and bakery goods
are considered to be the best products to fortify (Gawlik-Dziki et al.,
Antioxidant activity [mg g−1]a 2013). This study investigates whether enrichment of wheat dough with
ABTS•+ 593.9 ± 15.6
an onion skin extract enhances the total antioxidant activity of the final
DPPH• 513.3 ± 13.2
CUPRAC 441.4 ± 11.5 product. For the fortified bread samples, we determined the total
Total polyphenols content 543.7 ± 22.5 polyphenol content using Folin-Ciocalteu's method, and the antioxidant
Total flavonoids content 340.4 ± 7.05 properties using the DPPH and CUPRAC methods.
Phenolic compounds profile [mg g−1] Our research has shown that enrichment of wheat dough with an
Quercetin 315.6 ± 2.5
Kaempferol 10.9 ± 1.1
onion skin extract significantly increases the levels of polyphenols and
Isoramnethin 14.8 ± 3.1 the antioxidant activity of final product. We observed a closely corre-
Quercetin-3-glucoside 40.3 ± 2.5 lation between onion skin extract dose and antioxidant activity of en-
Basic chemical composition [g 100g−1] riched bread (p < 0.05). Antioxidant activity and phenolic compounds
Dry matter content 94.1 ± 4.7
content in bread increased with increasing extract dose. For instance,
Total carbohydrate 51.6 ± 2.6
Protein content 5.3 ± 0.2 the enrichment of the dough with a 0.5% of the extract caused an in-
Total dietary fiber 1.0 ± 0.1 crease in total phenolic content in bread by 4-times and antioxidant
Crude fat 0.0 activity by 7-times measured by the CUPRAC method (Fig. 4).
a
Park and Kim (2007) observed that the addition of a commercial
- as a quercetin equivalent.
grape seed extract (0.3 g, 0.6 g and 1 g) to 500 g of the flour to be used
to make bread also enhanced the antioxidant properties of the final
has many advantages, it is relatively expensive, considering the high
product. They noticed, however, that the enhancement was not pro-
cost of installing high-pressure equipment and of compressing the sol-
portional to the extract dose used. The authors supposed that their
vent.
extract was characterized by low stability for the baking conditions.
Wang, Zhou, and Isabelle (2007) in turn evaluated the quality of bread
enriched with a green tea extract at doses of 1.5 g and 5 g per 1 kg of the
3.2. Antioxidant properties of onion skin extract
flour to be used to make bread. Their research showed that the addition
of a green tea extract improves the general quality of bread but also
HPLC-UV-Vis/DAD analysis revealed that an onion skin extract is a
significantly affects the sensory characteristics of bread, inter alia, its
good source of polyphenols (Table 3). In the course of our research, we
colour. In our research, we also noticed a positive effect of this extract
identified 4 phenolic compounds belonging to the flavonoid group, i.e.
on the bread colour. Since the green tea extract is characterized by a
quercetin, quercetin-3-glucoside, kaempferol and isorhamnetin. We
light brown colour, bread enriched with it becomes yellow to brown in
found that quercetin aglycone was the dominant phenolic compound
colour, depending on the dose used.
and its content was as high as 315.6 ± 2.5 mg g−1. We also proved, by
typical spectrophotometric methods based on the measurement of the
degree of reduction of the molybdenum ions (VI) contained in Folin-
4. Conclusions
Ciocalteu's reagent and on the ability to form complexes of flavonoids
with aluminium ions (III), respectively, the total content of polyphenols
Extraction of antioxidant compounds from onion skin using me-
and flavonoids in the extract to be high. Moreover, onion skin extract is
thanol as a solvent was dependent on the applied batch extraction
a source of carbohydrates, proteins and dietary fibber.
conditions, i.e. temperature and process time, as well as the volume of
The research evidence to date shows that the chemical composition
extraction solvent in relation to the onion skin mass. The optimal
of onion skin extracts, including the polyphenolic compound profile,
conditions of the process, in which the maximum yield of antioxidant
depends on the extraction technique and the process conditions ap-
compounds extraction is achieved from the skin mass unit i.e. the re-
plied. For example, Campone et al. (2018) acquired bioactive com-
lation of the skin mass to the volume methanol equal to 1:30, process
pounds from yellow onion skin using supercritical carbon dioxide and
temperature amounting to 44 °C and time of process to 145 min.
ethanol as a factor enhancing the solubility of polar components. They
The obtained onion skin extract was characterized by high anti-
identified 17 flavonoid compounds in the extract they had obtained,
oxidant activity which shaped by high content of flavonoid compounds,
including quercetin, isorhamnetin, kaempferol and their derivatives,
mainly quercetin aglicone. Moreover, total antioxidant activity of
mainly glycosides and polymerized forms. They also found proto-
wheat bread enriched with onion skin extract was closely correlated
catechuic acid in the extract. Whereas Munir, Kheirkhan, Baroutian,
with the enrichment rate. The presented research gives the opportunity
Quek, and Young (2018), using supercritical water, isolated quercetin
to conduct further research on the effect of onion skin extract on the
only, which was probably due to the low stability of the other poly-
sensory quality, texture parameters of bread enriched with the extract
phenols under the established process conditions.
as well as rheological properties of the dough.

6
T. Piechowiak, et al.
Fig. 4. Antioxidant activity of bread enriched with onion skin extract. A – total
phenolic compounds, expressed as a gallic acid equivalent, B – antioxidant
activity against DPPH radicals, expressed as a trolox equivalent. C- antioxidant
activity assayed with CUPRAC method, expressed as a trolox equivalent. The
bars and error bars represent the means and standard deviation of the mean
(x‾ ± SD ) Means values with the same lower case are not statistically significant
according to the T-Tukey test (α = 0.05).
Conflicts of the interest statement
The authors whose names are listed immediately below certify that
LWT - Food Science and Technology 117 (2020) 108614
they have NO affiliations with or involvement in any organization or
entity with any financial interest (such as honoraria; educational
grants; participation in speakers’ bureaus; membership, employment,
consultancies, stock ownership, or other equity interest; and expert
testimony or patent-licensing arrangements), or non-financial interest
(such as personal or professional relationships, affiliations, knowledge
or beliefs) in the subject matter or materials discussed in this manu-
script.
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