Journal of Industrial and Engineering Chemistry 18 (2012) 360–363

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Journal of Industrial and Engineering Chemistry

journal homepage: www.elsevier.com/locate/jiec

Characterization of wheat bran oil obtained by supercritical carbon dioxide and

hexane extraction
Go-Woon Jung, Hee-Moon Kang, Byung-Soo Chun *
Institute of Food Science, Department of Food Science and Technology, Pukyong National University, 599-1 Daeyeon-3dong, Nam-Gu, Busan 608-737, Republic of Korea

A R T I C L E I N F O A B S T R A C T

Article history: Supercritical carbon dioxide (SC-CO2) and soxhlet extraction using was carried out to extract oil from
Received 21 January 2011 wheat bran oil. For SC-CO2, the pressure and temperature were ranging from 10 to 30 MPa and 313.15–
Accepted 12 April 2011 333.15 K. The extraction was performed in a semi batch process with a CO2 flow rate of 26.81 g/min for
Available online 10 November 2011
2 h. Wheat bran oil was characterized to investigate the quality. Acid value (AV) and peroxide value (POV)
were higher in hexane extracted oil compared to SC-CO2 extracted oil. Induction period was measured by
Keywords: rancimat test. The oil obtained by SC-CO2 extraction had higher capability to delay the oxidation by
Supercritical carbon dioxide
surrounding environment. The DPPH radical scavenging activity was also measured. The SC-CO2
Wheat bran oil
Antioxidant activity
extracted oil showed higher radical scavenging activity compared to hexane extracted oil.
Hexane ß 2011 The Korean Society of Industrial and Engineering Chemistry. Published by Elsevier B.V. All rights
reserved.

1. Introduction be avoided in a conventional separation method, since relatively

Wheat is one of the popular cereal grains. Wheat bran is a by-
product of the flour milling industry. Wheat bran obtained after
the milling of wheat grains. In recent years, several researchers
have studied about wheat bran [1]. Wheat bran is good sources of
diverse nutrients and antioxidant activity materials such as
tocopherols, carotenoids, phenolics and phytochemicals [2–5].
The role of dietary antioxidant materials is prevent to many human
diseases such as cancer, cardiovascular disease and other chronic
diseases [6,7]. Reactive oxygen species (ROS) produced through life
activities endogenously and these ROS interact with cellular
biomolecules and then oxidized to biomolecules such as lipids,
proteins and DNA. Eventually, oxidized biomolecules are likely to
evolve into a chronic disease. Oxidative metabolism continuously
produces oxygen-centered free radicals and other reactive oxygen
species in vivo, resulting in cell death and tissue damage [8]. The
role of antioxidant compounds is delay or prevent to oxidation of
cells [9]. Due to these properties, natural antioxidants are widely
used in food, cosmetic and pharmaceutical industries to improve
the quality for health promotion.
Separation of active ingredients from various materials (plants,
food-by-products, algae, etc.) has attracted much attention in food,
cosmetics and pharmaceutical industries. Conventionally, differ-
ent antioxidants have been obtained using organic solvents.
Decomposition or degradation of thermolabile compounds cannot
high temperatures are required for these processes. Moreover, the
use of organic solvent is not environmental friendly, and the
production is currently limited by safety and regulatory con-
straints to the concentration of toxic residues of conventional
solvents. Supercritical fluid extraction has been widely employed
as an alternative to organic solvent for the extraction of a variety of
compounds from different matrices [10–16]. Carbon dioxide (CO2)
is probably the most widely used supercritical fluid because of its
critical temperature (304.25 K), which makes it an ideal solvent for
extracting thermally labile materials. It is also excellent as a
solvent due to its inertness, non-toxicity, non-flammability and
low cost. CO2 is applied in supercritical fluid extraction processes
at near-environmental temperatures thus minimizing heat re-
quirement and thermal damage to bioactive compounds [17]. No
toxic residual solvent is remained in the final extracts [18]. Several
studies have been carried out in which supercritical carbon dioxide
(SC-CO2) extraction was used to produce high value oil with
different antioxidants [19–22]. Therefore, the aim of this study was
to check the quality of wheat bran oil obtained by SC-CO2
extraction and to compare with the oil obtained by organic solvent
extraction using hexane.
2. Experimental
2.1. Materials

The sample was provided from Young-Nam Flour Mills

* Corresponding author. Tel.: +82 51 629 5830; fax: +82 51 629 5824. Company (Busan, Republic of Korea). Carbon dioxide (99.99%
E-mail address: bschun@pknu.ac.kr (B.-S. Chun). pure) was supplied by KOSEM (Yangsan, Republic of Korea). 2,2-

1226-086X/$ – see front matter ß 2011 The Korean Society of Industrial and Engineering Chemistry. Published by Elsevier B.V. All rights reserved.
doi:10.1016/j.jiec.2011.11.094
 G.-W. Jung et al. / Journal of Industrial and Engineering Chemistry 18 (2012) 360–363
Diphenyl-1-picryl-hydrazyl (DPPH) was purchased from Sigma–
Aldrich (St. Louis, MO, USA). All other chemicals used in different
analysis were analytical or HPLC grade.
2.2. Sample preparation
After drying in an oven at low temperature, wheat bran samples
were crushed in a mechanical blender and sieved (500 mm) by a
mesh. The sieved samples were then stored at 275.15 K and used
for SC-CO2 extraction.
2.3. SC-CO2 extraction
A laboratory scale supercritical fluid extraction unit was used
for extracting oil from wheat bran (Fig. 1). The extraction apparatus
was able to be operated at maximum pressure of 30 MPa. The flow
rate of CO2 was 26.81 g/min and it was constant for all extraction
conditions entire over the extraction period of 2 h. The process of
extraction of wheat bran oil by SC-CO2 at different temperatures
(313.15–333.15 K) and pressures (10–30 MPa) has been discussed
in detail in our previous study [3].
2.4. Organic solvent extraction
The extraction was carried out in a soxhlet apparatus using
hexane as solvent. Five grams of sample was placed into the
extraction thimble and the extraction was run 12 h until the
colour of the condensed solvent at the top of the apparatus was
clear. After extraction, hexane was evaporated by a rotary
vacuum evaporator (EYELA, Japan) and the oil was collected in a
vial.
2.5. Characterization of oil
2.5.1. Acid value (AV)
The AV was assessed according to the method described
previously by Shao et al. [23]. Briefly, 1 g of sample was dissolved in
100 mL of ether and ethanol (1:1) solution. The AV of oil was
analyzed by titration with 0.1 N KOH–ethanol solution and
Fig. 1. Schematic diagram of supercritical extraction process.
361
phenolphthalein as an indicator. The AV was calculated using
the equation
V  c  56:1
AV ¼
m
where V is the volume of the KOH–ethanol solution of the titration
(mL), c is the concentration of the KOH–ethanol factor, m is the
weight of the sample (g), and 56.1 is the molecular weight of KOH.
2.5.2. Peroxide value (POV)
The POV was determined according to Wang and Lin [24].
Briefly, 1 g of sample was dissolved in 30 mL of chloroform and
acetic acid (2:3) solution. The POV of oil was analyzed by titration
with 0.1 N Na2S2O3 solution using an iodometry. The POV was
calculated using the equation
ðA  BÞ  F
POV ¼  10 ð0:01  1000 mg=kgÞ
c
where A is the volume of the 0.1 N Na2S2O3 solution for the
titration (mL) of sample, B is the volume of the 0.1 N Na2S2O3
solution for the titration (mL) of blank, F is the concentration of the
0.1 N Na2S2O3 factor and c is the weight of the sample (g).
2.5.3. Rancimat test for antioxidant activity
The rancimat method (Metrohm model 743, Herisan,
Switzerland) determined the induction periods by measuring
the increase in the volatile acidic by-products released from the
oxidizing oil at 383.15 K with an air flow rate 20 L/h. The
concentration of the degradation products, which are transferred
into distilled water, is assessed by measuring the conductivity.
Longer induction periods suggest stronger antioxidant activity for
the extracted oil.
2.5.4. DPPH radical scavenging activity
The DPPH radical scavenging effect was assessed according to
the method described Hsu et al. [25]. Aliquots of 1 mL methanolic
sample solution added 5 mL of freshly prepared 0.1 mM DPPH
362 G.-W. Jung et al. / Journal of Industrial and Engineering Chemistry 18 (2012) 360–363
methanolic solutions, in a total volume of 2 mL. After mixing
thoroughly, it was kept for 50 min in the dark place. The
absorbance of the reaction mixture was read at 517 nm with
spectrophotometer. For blank, 1 mL of the methanol was
replaced with the extracted oil. The percentage of free
radical scavenging effecth was A calculated i by the equa-
tionScavenging effect ð%Þ ¼ 1  A517 nm;sample  100
517 nm;blank
3. Results and discussion
3.1. SC-CO2 extraction
SC-CO2 extraction curves of wheat bran oil at different
temperatures (313.15–333.15 K) and pressures (10–30 MPa) have
been described in details in our previous work [3]. The highest oil
yield obtained at higher temperature and pressure. The variation of
applied pressures and temperatures greatly affected the oil
solvating power of SC-CO2 and, hence, the amount of yield.
3.2. Characterization of wheat bran oil
3.2.1. AV and POV
In the present study, storage properties of oils obtained by SC-
CO2 and organic solvent extraction were compared under constant
conditions. The extracted condition was 30 MPa and 333.15 K,
because the oil extracted under this condition has the highest
antioxidant activity in rancimat test. AV and POV were measured
the quality of the oil during the storage period of 20 days. Figs. 2
and 3 shows the AV and POV value of wheat bran oil obtained by
SC-CO2 (30 MPa and 333.15 K) and hexane extraction. The oil was
stored at 298.15 K. The AV reflects the amount of free fatty acid in
oil. The AV was used to determine the acidity of the oil. The AV and
POV were higher in hexane extracted oil compared to SC-CO2
extracted oil. It might be happened due to applying higher
temperature at organic solvent extraction. Lee et al. [26] also
reported similar results from mackerel viscera. The AV of hexane
extracted oil increased by oxidation process during storage of 20
days. On the other hand, the AV of SC-CO2 extracted oil decreased
after a certain period. The POV provides the oxidation state of the
lipids. This value was used to measure the rancidity that occurred
by auto-oxidation. The POV of oils extracted with organic solvents
rapidly increased up to 5 days of storage but in SC-CO2 extracted
samples it increased very slowly up to 15 days. The results of POV
Fig. 2. The AV of SC-CO2 and hexane extracted wheat bran oil.
indicated that oils extracted with SC-CO2 were effective in delaying
of initial oxidation process. The AV and POV are dependent on the
nature of processing and the factors such as the surrounding
moisture, air, and temperature.
3.2.2. Antioxidant activity measurement by rancimat test
The rancimat method is commonly used to evaluate the
antioxidative properties of various antioxidants and is based on the
increase of electrical conductivity due to the formation of volatile
dicarboxylic acids as a result of lipid oxidation. Rancimat machine
was used to measure the induction period. The induction period
provides the starting point of oxidation of lipid. Fig. 4 shows the
induction period of wheat bran oil obtained by SC-CO2 extraction.
The oil obtained by hexane extraction showed the induction period
of 4.22 h. Therefore, it was found that the induction period of SC-
CO2 extracted oil was higher than that of hexane extracted oil. It
indicated that SC-CO2 extracted oil had more efficiency to delay the
oxidation in unfavorable environments.
It was observed that the induction period was longer at higher
pressure and temperature. The oil extracted at 30 MPa and
333.15 K has the longest induction period of 16.66 h. This result
indicated that the oil extracted at high temperature and pressure
had higher efficiency to delay oxidation. It might be happened due
to presence of high amount of antioxidants at higher temperature
and pressure. The solvating power of SC-CO2 increased at higher
pressure and temperature that enhanced the extraction of
antioxidants. This finding agreed with our previous results [2,3].
3.2.3. DPPH free radical scavenging effect
The DPPH radical scavenging effects of wheat bran oils obtained
by SC-CO2 and hexane extraction are shown in Table 1. Wheat bran
oils with SC-CO2 showed appreciable free radical scavenging
activities. The oil obtained by SC-CO2 extraction at 30 MPa and
333.15 K showed the strongest radical scavenging activity (94.8%)
compared to the oil obtained at other extraction conditions and
also organic solvent. The DPPH radical scavenging effect of oil
obtained by organic solvent was 74.79%. In general, the effect of
pressures on DPPH radical scavenging activities was dominant
than the temperatures. No significant difference of the DPPH
radical scavenging activities of wheat bran oil obtained by SC-CO2
at different temperatures was found. Zhou and Liangli [27] also
reported the almost DPPH scavenging activity of wheat bran
extract using organic solvents. Therefore, such dietary antioxidants
Fig. 3. The POV of SC-CO2 and hexane extracted wheat bran oil.
 G.-W. Jung et al. / Journal of Industrial and Engineering Chemistry 18 (2012) 360–363 363

Table 1
DPPH free radical scavenging activity of wheat bran oil obtained by SC-CO2 and hexane extraction.

Temperature (K) DDPH free radical scavenging effect (%)

SC-CO2 Hexane

10 MPa 15 MPa 20 MPa 25 MPa 30 MPa

313.15 69.13 72.12 85.77 78.02 93.20 74.79

323.15 69.72 73.29 87.67 78.80 94.00
333.15 70.03 75.51 89.32 80.49 94.80

extraction from different species cultivars will be more effective

for human and other living organisms.

Acknowledgement

This research was financially supported by a grant from the

Ministry of Knowledge and Economy, Korea Industrial Technology
(KOTEF) through the Human Resources Training Project for
Regional Innovation.

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Fig. 4. Induction period of wheat bran oil at different extraction conditions at
rancimat test.
from wheat bran may be particularly important in fighting these
diseases by conferring protection against free radical damage to
cellular DNA and lipids.
4. Conclusions
Wheat bran oil was extracted by SC-CO2 extraction at different
pressures and temperatures. In this study, wheat bran oil was
characterized in terms of quality such as AV and POV and
antioxidant capability. The quality of SC-CO2 extracted oil was
better compared to hexane extracted oil due to lower AV and POV.
In rancimat test, wheat bran oil extracted by SC-CO2 had longer
induction period than that obtained by hexane. The result of DPPH
radical scavenging effect was excellent in case of SC-CO2 extracted
oil. Therefore, the oil with good quality obtained by SC-CO2
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