Food Sci. Biotechnol.
23(2): 615-621 (2014)
DOI 10.1007/s10068-014-0084-6
RESEARCH ARTICLE
Antioxidant Activities of Onion (Allium cepa L.) Peel Extracts
Produced by Ethanol, Hot Water, and Subcritical Water Extraction
Kyoung Ah Lee, Kee-Tae Kim, Hyun Jung Kim, Myong-Soo Chung, Pahn-Shick Chang, Hoon Park, and
Hyun-Dong Paik
Received July 29, 2013; revised September 17, 2013; accepted September 23, 2013; published online April 30, 2014
© KoSFoST and Springer 2014
Abstract Onion (Allium cepa L.) peels were extracted by
ethanol, hot water and subcritical water (SW) extraction
and their antioxidant activities were evaluated. Extraction
yields of SW extraction were 4-fold higher than ethanol
extraction. However, the ethanol extraction increased the
total phenolics contents (327.5 mg GAE/g extract) and
flavonoids contents (183.95 mg QE/g extract) in the onion
peel extract. The onion peel extracts by ethanol extraction
showed greater DPPH radical scavenging activities and
greater antioxidant activities determined by ferric thiocyanate
assay than those by hot water extraction and SW extraction
at 165oC. Antioxidant activity of onion peel extract by SW
extraction at 110oC was similar to that of ethanol extraction.
HPLC profiles revealed that SW extraction at lower
temperature (110oC) increased the concentration of
quercetin. These results demonstrated that the onion peel
Kyoung Ah Lee, Hyun-Dong Paik (
)
Department of Food Science and Biotechnology of Animal Resources,
Konkuk University, Seoul 143-701, Korea
Tel: +82-2-2049-6011; Fax: +82-2-455-0381
E-mail: hdpaik@konkuk.ac.kr
Kee-Tae Kim, Hyun-Dong Paik
Bio/Molecular Informatics Center, Konkuk University, Seoul 143-701,
Korea
Hyun Jung Kim
Department of Food Bioengineering, Jeju National University, Jeju 690-
756, Korea
Myong-Soo Chung
Department of Food Science and Engineering, Ewha Womans University,
Seoul 120-750, Korea
Pahn-Shick Chang
Department of Agricultural Biotechnology, Seoul National University,
Seoul 151-742, Korea
Hoon Park
Department of Food Science, Sun Moon University, Chungnam 336-708,
Korea
extracts produced by SW extraction technique have great
potential as a source for useful antioxidant.
Keywords: onion peel, antioxidant activity, subcritical
water extraction, flavonoid, quercetin
Introduction
Onion (Allium cepa L.), one of the most frequently
consumed vegetables, is known to have many health
benefits because of its flavonoids contents. Onion extracts
can function as potent cardiovascular and anticancer agents
that exert hypocholesterolemic, thrombolytic, and antioxidant
effects (1-4). Flavonols, which are primarily detected as
glycosides of quercetin and kaempferol in onions, have
been identified as major contributors to the antioxidative
activities and health benefits (5,6). Orange colored onion
peel including several phenolic compounds, such as
quercetin, gallic acid, ferulic acid, and kaempferol (7). The
major flavonoid in the onion peel is quercetin, which is
present in one of its conjugated forms, as quercetin 4'-O-β-
glycopyranoside, quercetin 3,4'-O-β-diglycopyranoside, and
quercetin 3,7,4'-O-β-triglycopyranoside (6). The onion
ranked the highest in quercetin content among 28
vegetables and 9 fruits (8), and its flavonoid compounds
are mostly present in the onion peels at levels significantly
higher than in the edible portion of the onion (9).
Conventional extraction processes based on organic
solvents have been previously applied to extract natural
antioxidants (10-12). There has been a recent increase in
the use of environmental clean technologies capable of
providing high-quality and high-activity extracts, while
precluding any toxicity associated with the solvents. The
subcritical water (SW) extraction technique is an friendly
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environmental method with a broad range of applications,
including the extraction, hydrolysis, and wet oxidation of
organic compounds (13,14). The SW extraction system
involves water with high temperatures ranging from 100oC
to 374oC, under sufficiently high pressure to maintain the
water in its liquid state. Under subcritical conditions, the
dielectric constant of water (polarity) can be lowered to
that of ethanol or methanol, thus selectively extracting
different classes of compound (15).
Therefore, the SW extraction as a friendly environmental
process can be used for the extract of functional compounds
from onion peel for safe and rapid methodology. The
objective of this study were to determine the optimal
conditions for the extraction of phenolic compounds
including flavonoids from onion peel by SW extraction
process and to evaluate its antioxidant activities in
comparison with traditional processes, ethanol extraction
and hot water extraction.
Materials and Methods
Onion peel Onions cultivated at Ui-Seung, Gyeongsang
Buk-do, South Korea were purchased from a local market.
Only the orange-colored onion peels were used for the
extraction of polyphenol compounds. The peels were dried
in a drying oven at 60oC for 10 h until the moisture
contents were 4%-5%(w/w). The dried onion peels were
then ground to a particle size of 1-10 mm using a high-
speed mixer (Blender 7012S; Waring, Torrington, USA),
and were stored at 4oC until used.
Preparation of onion peel extracts For ethanol
extraction, onion peels (20 g) was mixed with 200 mL of
70%(v/v) ethanol and heated for 3 h at 60oC in a water
bath. For hot water extraction, the onion peel powder was
dissolved in 200 mL distilled water and heated for 3 h at
80oC in a water bath. After ethanol and hot water extraction,
the slurry was filtered through filter paper (Whatman No.
2; Whatman, Kent, UK) and the solid residue was
extracted more than twice. After the solvent was
evaporated by using a rotary evaporator (EYELA N-
1000V; Tokyo, Japan), the extract was transferred to a
freeze-drying tube and lyophilized. The dried extracts were
then weighed and stored at 20oC prior to analysis.
Subcritical water extraction was performed to extract onion
peels by using a subcritical extractor system (DIONEX
ASE 100; Dionex Corporation, Sunnyvale, CA, USA). The
process was carried out in a stainless steel cell. The cell
was filled with the mixture of onion peel powder and
diatomaceous earth (1:3 ratios) and distilled water. The
extraction temperature and static time were set at 110oC or
165oC for 15 min. The pressure was maintained less than
Lee et al.
500 psi. After 15 min, the fresh distilled water is pumped
through the entire pathway including the extraction cell for
washing. The system was then purged with nitrogen gas.
The SW extracts were freeze-dried and stored at 20oC until
used (15).
Determination of total phenolics and flavonoids
contents in onion peel extracts Total phenolics contents
were determined by using Folin-Ciocalteau assay (16). The
onion peel extracts were dissolved in methanol (1 mg/mL)
and an aliquot (100 µL) was mixed with 2 mL of 2%
aqueous sodium carbonate solution. After 3 min, 100 µL of
50% Folin-Ciocalteau’s reagent (Sigma-Aldrich Chemical
Co., St. Louis, MO, USA) was added to the mixture. After
30 min of standing, the absorbance was measured at 750
nm with a spectrophotometer (2120 UV; Optizen, Daejeon,
Korea). Gallic acid (99%, Tokyo Chemical Industry Co.,
Ltd.) was used as a calibration standard and the results
were expressed as mg of gallic acid equivalent (GAE) per
gram of extract (dry weight). The total flavonoids contents
were determined by aluminum chloride colorimetric method
(17). The onion peel extracts were dissolved in methanol
(1 mg/mL) and solution (0.5 mL) were mixed with 1.5 mL
of 95% ethanol, 0.1 mL of 10% aluminum chloride, 0.1
mL of 1 M potassium acetate, and 2.8 mL of distilled
water. After incubation at 25oC for 30 min, the absorbance
was measured at 415 nm. Quercetin (HPLC grade; Sigma-
Aldrich) as a standard was used to construct the calibration
curve and the results were expressed as mg of quercetin
equivalent (QE) per gram of extract (dry weight).
Determination of quercetin by HPLC The concentration
of quercetin in onion peel extracts produced by ethanol, hot
water, and SW extraction, were determined by HPLC (18).
The onion extract was dissolved in 50% dimethyl sulfoxide
(DMSO, 99%; Sigma-Aldrich) in methanol (5 mg/mL).
After filtering through a 0.45 µm membrane filter, 20 mL
was injected to an Agilent 1100 HPLC (Agilent Technologies,
Palo Alto, CA, USA). The separation carried out on a
DuPont Zorbax ODS C18 column (150×4.6 mm, i.d. 5 µm).
The mobile phase was composed of solvent A (0.1%
formic acid (v/v), pH 2.7, HPLC grade; Sigma-Aldrich)
and solvent B (100% acetonitrile, HPLC grade; J.T.Baker,
Phillipsburg, NJ, USA). Linear gradient program at a flow
rate of 0.5 mL/min was performed as follows: 0-10 min,
linear gradient 10 to 20% solvent B; 10-20 min, isocratic
20% solvent B; 20-25 min, linear gradient 20 to 30%
solvent B; 25-35 min, linear gradient 30 to 33% solvent B;
35-40 min, linear gradient 33 to 40% solvent B; 40-60 min,
linear gradient 40 to 60% solvent B. The peaks were
identified using UV absorbance at 254 nm and quercetin
was used as a standard flavonoid.