Journal of Industrial and Engineering Chemistry 17 (2011) 537–542

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Journal of Industrial and Engineering Chemistry

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

Synergetic and antagonistic role of natural antioxidant in the autoxidation of

soybean oil
Kyu-Wan Lee a,*, Jing Li a, Young-Wun Kim b, Keun-Woo Chung b, Yoon Jin Lee c, Han Bin Oh c
a
Division of Biological and Chemical Engineering, Yanbian University of Science & Technology, Yanji, Jilin 133000, China
b
Lubricant Team, KRICT, Daejeon 305-600, South Korea
c
Department of Chemistry, Sogang University, Seoul 121-742, South Korea

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

Article history: The antioxidant effects of ascorbic acid (vitamin C), a-tocopherol (vitamin E), 2,6-di-t-butyl-4-cresol
Received 14 May 2010 (BHT) and 4-hydroxy-2,2,6,6-tetramethyl-piperidine-N-oxyl (abbr. N-oxyl = 4-hydroxy-TEMPO) on the
Accepted 25 October 2010 autoxidation of soybean oil (SBO) were evaluated at 140 8C in four different concentrations (at 1.0, 0.7,
Available online 28 April 2011
0.5, and 0.3 wt% of SBO). Ascorbic acid and a-tocopherol showed lower active oxygen concentration at
the range of 0.5–0.7 wt% concentrations than that at 1.0 wt%. However, on the contrary, 2,6-di-t-butyl-4-
Keywords: cresol and N-oxyl showed higher effect with increasing concentration. In 1.0 wt% antioxidant
Soybean oil
concentration, the inhibiting effect showed in the following order, 2,6-di-t-butyl-4-cresol > a-
Antioxidant
tocopherol > N-oxyl > ascorbic acid and in lower concentration of 0.5 wt%, the order was inverted as
Ascorbic acid
a-Tocopherol follows: ascorbic acid > a-tocopherol > 2,6-di-t-butyl-4-cresol > N-oxyl. The inversion of the order at
Hydrogen bonding lower concentration could be attributed to the synergetic and antagonistic effect arousing from
Synergetic/antagonistic role hydrogen bonding. The composition of oxidized triglyceride was investigated by means of LC/ESI-MS/MS
spectroscopy.
ß 2011 The Korean Society of Industrial and Engineering Chemistry. Published by Elsevier B.V. All rights
reserved.

1. Introduction
Soybean oil, an acyltriglyceride, one of the most popular
vegetable oils, is an ingredient widely used in cooking and also in
the food industry [1]. SBO contains many unsaturated fatty acids
such as oleic, linoleic, and linolenic acids so that it can be easily
oxidized by oxygen which has a triplet electronic configuration in
ground state [2].
Therefore SBO has a-hydrogen, allylic, bisallylic hydrogen,
methylene hydrogen and methyl groups. In general, double bonds
present in fatty acids can offer several possibility to chemically
modify the structure to improve some of their properties. One
example is a hydrogenation of double bond [3] for solidification of
SBO. Epoxidation is another example of widely used modification.
Epoxidized soybean oil (ESBO) is commercially available and is
used as a plasticizer for polyvinyl chloride [4] and can further be
used to a new biocomposite [5,6] because SBO and its epoxide are
cross linked with mono-, and di-amine [5].
From the food and health-related standpoint, the presence of
double bonds in fatty acids, however, are troublesome. They are
vulnerable to oxidation even when stored at low temperatures and
* Corresponding author.
E-mail address: kuiwanlee@hanmail.net (K.-W. Lee).
used in kitchen as frying oil which often renders SBO more prone to
rancidity and quality decline for food use (autoxidation or lipid
peroxidation) [7]. Therefore, many studies were carried out for the
stabilization of substrates, and similar effects of a- and g-
tocopherol on formation of hydroperoxide from methyllinoleate
[8], alkyl substituted ascorbic acids as free radical quenchers [9],
and Icariin as antioxidant against free radical induced peroxidation
of linoleic acid [10].
In China, Japan and Korea, the vegetable oil is used mainly for
frying and cooking at high temperature often to the boiling
temperature. Therefore the interacting behaviors between cooking
oil and antioxidant could be of interest to investigate. Also
autoxidations at the high temperature and high pressure were well
studied and industrialized [7,11]. Among them are cumyl process
(90–130 8C, 5–10 atm) to produce acetone and phenol, and
cyclohexane oxidation (125–165 8C, 8–15 atm) to cyclohexanone
and cyclohexanol (so called KA oil), ethylene oxide production (250–
300 8C, 10–20 atm), maleic anhydride from butene (350–450 8C, 2–
3 atm) and/or benzene (400–450 8C, 2–5 atm), and many others.
In the food industry, extensive efforts have been made towards
preventing or minimizing oxidation of SBO. In this regard, an
antioxidant is very attractive due to its capability of slowing or
preventing the oxidation of other molecules. Antioxidant is now
widely accepted as a supplementary dietary ingredient. In
medicine, it is also reported to have a potential efficacy in

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.2010.10.015
538 K.-W. Lee et al. / Journal of Industrial and Engineering Chemistry 17 (2011) 537–542
preventing diseases such as inflammation, cancer and diabetes
[12]. Antioxidants have been used as preservatives in the food and,
cosmetics industry, and preventing for the degradation of rubber
and petroleum fuels.
Antioxidants are also called inhibitors, since they slow down or
inhibit the oxidation of substrates having double bonds [13].
Antioxidants are often categorized into two classes, i.e., radical
trapping and peroxide decomposer based on their mechanism in
preventing oxidation [14]. Phenols and secondary aromatic amines
are well known as the radical traps and phosphate are known to be
a good peroxide decomposer.
In the present study, we have investigated the antioxidants
effect such as ascorbic acid and a-tocopherol, compared with 2,6-
di-t-butyl-4-cresol (BHT) and 4-hydroxy-2,2,6,6-tetramethyl-pi-
peridine-N-oxyl (4-hydroxy-TEMPO) at different concentrations
(1.0, 0.7, 0.5, and 0.3 wt%). The two vitamins are well known as
powerful radical capturer in human body. It was of special interest
to know, the role of ascorbic acid, which has four hydroxyl groups
in comparison with petrochemical inhibitors like BHT and 4-
hydroxy-TEMPO.
In the previous work of Lee et al. [15], only the inhibiting effect
of the phenol derivatives was investigated. In this work, the
inhibiting effect of some natural alcohols, e.g., ascorbic acid and a-
tocopherol were added to obtain further information by comparing
with petrochemical antioxidants.
The authors also investigated the reaction phenomena in
different concentrations of natural inhibitors and identified the
oxidation products and the composition of SBO by means of LC–
ESI-MS/MS.
2. Experimental
2.1. Materials and reaction
2.1.1. Soybean oil purification [3]
In order to eliminate interfering water, coloring agent, and
antioxidant from the purchased soybean oil, (Zhonglyang group,
Beijing, China. Fortune trade mark), the oil was purified by the
following procedures. Into a 1 L three necked flask which equipped
with a Dean Stark separator, water cooling condenser and
mechanical stirrer, a mixture of 800 g SBO and 40 g of active
carbon was added and refluxed (160–180 8C) for about 1 h with
constant stirring. The mixture was filtered while still warming.
About 2.0 mL of water was collected. The purified SBO has acid
value of 0.165–0.270 mg KOH (Chungsan Chemical Co., CSS-J 520)
[16], iodine value of 130 (ASTM D 1951), color index of 1.2 (ASTM D
1500) and the pH measured directly with a pH meter.
The active oxygen concentration of purified soybean oil was
negligible.
2.1.2. Autoxidation of pure SBO [15,17]
Into a 200 mL three-necked flask equipped with an oxygen inlet
purging tube, a condenser, a magnet bar and 100 g of purified SBO
were placed. It was set in a silicon oil bath which is equipped with a
mechanical stirrer, and a thermocoupled heating plate (Daihan
Scientific, Seoul, Korea, WiseStir). When the temperature was
reached to the set temperature, oxygen gas was flowed at a rate of
1.11 mL/g min (111 mL/min) through the flow meter (Dwyer
Instruments, Inc., MI, USA, Rate-Master). In this experiment, pure
oxygen gas (purity: 99.999%) was used as oxidizer.
2.1.3. Autoxidation of SBO in the presence of antioxidants
The inhibitor was used along with 0.3, 0.5, 0.7 and 1.0 wt% of
substrate. During the oxidation process, in different time interval
and at different temperature, 0.3–0.5 g of oxidized SBO was taken
to analyze the active oxygen and to use for the instrumental
analyses. Each sample was titrated by iodometry to quantify the
amount of active oxygen.
In this experimental, ascorbic acid (Aldrich, >99%, mp 193 8C),
a-tocopherol (Aldrich 97%), 2,6-di-t-butyl-4-cresol (TCI, >99%, mp
71 8C) and 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (4-
hydroxy-TEMPO, TCI > 98%, mp 69–71 8C) were used without
further purification as antioxidant.
2.1.4. Iodometry [18]
For the quantitative analysis of active oxygen of SBO
hydroperoxide, 0.3–0.5 g of oxidized SBO was added to a flask
where a 20 mL of glacial acetic acid was placed. After that, 4 mL of
saturated KI solution was added into the flask and was left to
decompose the remaining peroxide in a water bath maintaining
82 8C for 5 min. Then added 50 mL distilled water and with
frequent shaking, added 2–3 drops of starch indicator and the
isolated iodine was titrated with 0.1 mol/L Na2S2O3 solution under
constant stirring.
2.1.5. Liquid chromatography–electrospray ionization tandem mass
spectrometry (LC–ESI-MS/MS) [19]
All LC–ESI-MS/MS experiments were carried out on an ion trap
mass spectrometer (LCQ Deca, Thermo Scientific, Waltham, MA,
USA) coupled with an Agilent 1200 series HPLC system (Agilent
Technologies, Santa Clara, CA, USA). The mass spectrometer was
operated in the positive ion mode. The experimental parameters
were normally set to the followings: spray voltage, 6.50 kV; capillary
temperature, 178 8C; capillary voltage, 40 V; tube lens off-set
voltage, 35 V; sheath gas flow rate (arb), 0; normalized collision
energy, 20–25%; isolation width, 1.5 Da. The chromatographic
separation was achieved on a Zorbax ODS analytical column (C18,
4.6 mm  250 mm, 5 mm; Agilent Technologies, Palo Alto, CA, USA)
at room temperature. Mobile phase A and B were an HPLC grade
methanol (0.1% ammonia/formic acid; pH 5.3) and chloroform
(HPLC grade, Sigma, Seoul, Korea), respectively. A binary gradient at
a flow rate of 1 mL/min was applied using an HPLC pump. The
following eluent gradient was applied: 0–5 min, 95% eluent A; 5–
25 min, 80% eluent A; 25–35 min, 65% eluent A; 35–38 min, 5%
eluent A. Before each HPLC run, the column was equilibrated.
3. Results and discussion
3.1. Autoxidation of SBO in the presence of different concentration of
antioxidants
Figs. 1 and 2 showed the active oxygen concentration in the
different concentration of antioxidants, ascorbic acid and 2,6-di-t-
butyl-4-cresol (BHT). Also, the behavior of a-tocopherol was similar
to ascorbic acid while the behavior of N-oxyl was similar to that of
2,6-di-t-butyl-4-cresol. In general, active oxygen concentrations
obtained with the addition of antioxidant show time-profiles
similar to that obtained for pure SBO. At all antioxidant concentra-
tions, the amount of active oxygen initially increased to a maximum
and then showed a decrease. More importantly, as clearly shown in
Figs. 1 and 2, the presence of an antioxidant generally induced a
decrease in the amount of active oxygen within SBO, as a result of
the antioxidation effect. As shown in the figures, the active oxygen
concentrations in the presence of antioxidants are lower than that of
pure SBO oxidation (maximum AO = 0.35% at about 170–180 min),
meaning that the used four chemicals act as the antioxidant.
In Fig. 1 shows the active oxygen concentrations at four
different concentrations of ascorbic acid (1.0 (^), 0.7 (&), 0.5 (~),
and 0.3 (*) wt% of ascorbic acid), wherein the percentage
concentrations of active oxygen are significantly less than that
obtained without the addition of antioxidant. The measurements
of the percentage concentrations of active oxygen with the
[(Fig._1)TD$IG] K.-W. Lee et al. / Journal of Industrial and Engineering Chemistry 17 (2011) 537–542 539

Table 1
pH at different medium and antioxidant concentration.

Antioxidant Ascorbic acid a-Tocopherol BHT

Medium SBO SBO* SBO SBO* SBO SBO*

0.3 (wt%) 2.37 2.88 4.66 2.78 3.74 5.12

0.5 (wt%) 2.91 3.67 4.73 3.13 3.62 5.21
0.7 (wt%) 3.11 3.32 4.84 3.31 –
1.0 (wt%) 3.32 3.53 4.93 2.91 3.47 5.38

SBO* = oxidized SBO.

Table 2
Acid value of the product in the presence of different antioxidant concentration.

Conc. (wt%) Ascorbic acid a-Tocopherol BHT N-oxyl

0.3 3.20 3.00 2.99 3.15

0.5 2.30 2.32 2.75 2.91
0.7 2.62 2.57 – –
1.0 2.95 2.78 2.31 2.82
Fig. 1. Change of AO concentration according to the running time in the presence of
ascorbic acid at 140 8C. ^: 1.0, &: 0.7, ~: 0.5, and *: 0.3 wt%. AV of SBO = 0.208 mg KOH/g.

leading to a synergetic effect. On the contrary, in high concentra-

addition of four different amounts of ascorbic acid also indicate
that there exists an optimal amount of ascorbic acid for the
maximal antioxidation effect. In the low concentration of 0.3 wt% a
peak appeared at 200 min just as pure SBO oxidation does. But a
paradoxical behavior was appeared, namely the active oxygen
concentration of 0.5 and 0.7 wt% was lower than that of 1.0 wt %
entry (see Fig. 1). This phenomenon appeared also in the case of a-
tocopherol.
In the comparison of a- and g-tocopherol [20], the superiority of
a-tocopherol was explained as the easy liberation of hydrogen of
alpha isomer than gamma isomer based on hyperconjugation and
steric effect of the structures. In that case, paradoxical phenomenon
was appeared also, but the authors could not explain the reasons.
The ascorbic acid and a-tocopherol showed superior effect than
BHT and N-oxyl in the range of 0.5–0.7 wt% concentration.
In our earlier work [15], the BHT showed better effect than 2,6-
dimethylphenol. Namely the more resonance forms are possible,
the more stable free radicals were formed, and showed reasonable
results where lower active oxygen concentration was observed in
higher antioxidant concentration, and no inversion.
The better activity of ascorbic acid in the concentration of 0.5–
0.7 wt% than in 1.0 wt% may be attributable to the competition
between hydrogen donating power and intra- and/or inter-
hydrogen bonding power of the ascorbic acid at different pH
[(Fig._2)TD$IG]
tion the equivalent hydrogen may be reacted according to the
oxidation rate and/or the extra or excess hydroxyl groups form
intra- and/or inter-hydrogen bond. The bonded hydrogen could not
take part in the reaction as inhibitor leading to the antagonistic
effect. In 0.3 wt%, the concentration is too low to function properly
but only acts as 1.0 wt%. The moderate concentration of ascorbic
acid in the given reaction condition was 0.5 wt%. Therefore the
authors measured the pH at different concentrations of ascorbic
acid, a-tocopherol and BHT in pure and oxidized soybean oil.
The measured results are summarized in Table 1. In each case,
the lower concentration showed lower pH in pure SBO as expected,
which indicates that in the high concentration the hydroxyl groups
will be formed the inter- and/or intra-hydrogen bonds and further
may be with double bonds and/or carbonyl groups of the oil but in
oxidized medium ascorbic acid showed the highest pH at 0.5 wt%
concentration and a-tocopherol also showed similar tendency at
0.7 wt%, because the equivalent protons were consumed as
antioxidant. But on the contrary, the BHT showed opposite
phenomenon, namely in SBO the pH decreased according to
increasing concentration. In oxidized product at 1.0 wt% the pH
showed highest pH because the equivalent proton of BHT was
consumed in the reaction as ascorbic acid at 0.5 wt% and a-
tocopherol at 0.7 wt% did. The different tendencies between
ascorbic acid and BHT can be attributed to the number of hydroxyl
group.
To reinforce the pH data we also measured the total acid value
of each product.
As shown in Table 2, the acid values (AV) well agreed with the
pH and inhibiting effect, namely the acid values of the products in
ascorbic acid and a-tocopherol were lowest at 0.5 wt% but in the
cases of BHT and N-oxyl were at 1.0 wt%. It means that ascorbic
acid and a-tocopherol played better inhibiting effect than
synthetic antioxidants. In the case of ascorbic acid, the acid values
well agreed with Fig. 1. On the contrary, in the case of synthetic
antioxidants BHT and N-oxyl showed reasonable results; the lower
antioxidant concentration is, the higher acid value obtains.

3.2. The composition of used oxidized soybean oil in different

antioxidant

In Table 3 are summarized the reaction results. As shown in Table

3, ascorbic acid showed the best inhibiting effect (only 4.5 area% of
TAG oxide) and the worst one was BHT (22.9 area% of TAG oxide).
Fig. 2. Change of AO concentration according to the running time in the presence of Figs. 3 and 4 show LC base peak chromatograms [19] acquired
BHT at 140 8C. ^: 1.0, ~: 0.5, and *: 0.3 wt%. for the SBO products oxidized in the presence of 1.0 wt% of ascorbic
540 K.-W. Lee et al. / Journal of Industrial and Engineering Chemistry 17 (2011) 537–542

Table 3 (M+H)+ at m/z 875.5, and its diacylglycerol products were observed
Composition of the products in the presence of each antioxidant.
at m/z 595.3 and 597.3. From the mass values of the precursor ion
Type of product Ascorbic acid (%) a-Tocopherol (%) N-oxyl (%) BHT (%) and of these fragments, it is possible to deduce that the molecular
DAG 0.4 0.5 0.3 0.7 species at m/z 892.4 correspond to a triacylglycerol molecule with
DAG oxide 0.0 0.0 0.0 0.1 18:2/18:3/18:3 sn chains. It is of note that from the MS/MS
TAG 95.1 88.2 80.8 76.3 spectrum of Fig. 5, it was not possible to assign the positions of its
TAG oxide 4.5 11.4 18.9 22.9 constituting fatty acid chains. This approach was repeatedly
applied for the identification of main peaks in Figs. 3 and 4.
The main components found in LC–MS/MS for autoxidized SBO in
acid and BHT, respectively. To investigate the whole components of the presence of four different antioxidants were triacylglycerols,
diacylglyceride, triacylglyceride, and their oxides in detail, which are listed in Table 4 and the confirmed compounds are
magnified 25 times the small peaks between 5 and 25 min as designated as (j, LLL), (k, LLO), (l, OOL) and (m, OOO). In them, the
shown in Fig. 4. components under 0.1 area% were deleted, for example, SOO, AOS,
The molecular species appearing in Figs. 3 and 4 could be BOL and BOS. And the confirmed diacyglycerides are (c, LL) (d, LO), (e,
identified with MS/MS. For example, a base peak appearing at OO) and (f, OP) in Fig. 4. The total oxidized products of the
31.6 min in Fig. 3 was subjected to MS/MS and the resulting MS/MS triacylglyceride are summarized in Table 5. The percentage of each
spectrum is shown in Fig. 5. Collisional activation upon the component was calculated as the base of 100 of the amount in the
precursor molecular species at m/z 892.4, which is cationized with Table 3. The oxidation in the presence of chemical antioxidants, 4-
NH4+, i.e., (M+NH4)+, led mainly to the release of ammonia, i.e., hydroxy-2,2,6,6-tetramethyl-piperidine-N-oxyl (4-hydroxy-TEM-
[(Fig._3)TD$IG]

Fig. 3. Liquid chromatogram of oxidized products in the presence of ascorbic acid.

[(Fig._4)TD$IG]

Fig. 4. Liquid chromatogram of the oxidized products in the presence of BHT.

[(Fig._5)TD$IG] K.-W. Lee et al. / Journal of Industrial and Engineering Chemistry 17 (2011) 537–542 541

Fig. 5. MS/MS spectrum for the peak at 31.6 min in Fig. 3.

Table 4
Composition of the triacylglycerides in the presence of each antioxidant.

TAG M.W. NH4+ form TAG-VC (%) TAG-VE (%) TAG-N-oxyl (%) TAG-BHT (%)

PLP 830 848 2.03 2.71 3.80 4.23

POP 832 850 0.40 0.59 0.69 0.84
LnLP 854 872 11.64 12.82 13.81 15.01
PLL 856 874 8.60 10.80 13.93 14.91
PLO 858 876 2.64 3.26 3.53 4.02
POO 860 878 0.29 0.31 0.40 0.41
LLLn 876 894 12.07 6.79 3.33 2.12
LLL 878 (j) 896 23.96 21.2 16.36 13.79
LLO 880 (K) 898 17.73 18.56 18.67 19.0
OOL 882 (l) 900 14.51 16.49 18.90 18.77
OOO 884 (m) 902 4.11 4.54 4.69 5.09
SOO 886 904 0.80 0.82 0.88 0.87
GLL 908 926 0.33 0.30 0.26 0.26
GLO 910 928 0.25 0.21 0.20 0.17
GOO 912 930 0.17 0.13 0.16 0.10
AOO 914 932 0.13 0.13 0.15 0.10
BOL 940 958 0.14 0.09 0.08 0.10

Total 100 100 100 100

Note: The order of three sn chains could not be determined in the MS/MS results.
Abbreviations: P, C16:0; Ln, C18:3; L, C18:2; O, C18:1, S, Stearic, C18:0; G, Gadoleic, C20:1; A, Arachidic, C20:0; B, Behenic, C22:0.

PO) gave about 18.9 area% of oxidized product and that in the
presence of 2,6-di-t-butyl-4-cresol showed more 22.9 area%, but the
natural antioxidants gave very low content, especially ascorbic acid
gave only 4.5 area% but a-tocopherol 11.4 area%. The stable free
radical compound, 4-hydroxy-TEMPO, had the minor values than
that of BHT. The authors expected that 4-hydroxy-TEMPO would be
the most effective one because of its stable free radical form. The
reason can be attributed to the oxidation of 4-hydroxy-TEMPO into
4-oxo-2,2,6,6-tetramethylpiperdine-N-oxide, or the free radical of
4-hydroxy-TEMPO easily abstracts hydrogen radical from SBO
which has many labile C–H bonds to form the piperidine, >N–H,
through >N-hydroxy, so that the concentration of 4-hydroxy-
TEMPO was remarkably decreased showing antagonistic effect as
antioxidant.
Some diacylglycerols were also observed, but with much lower
abundances (see Table 3). It is doubtful whether some of the
diacylglycerol were originated from the triacylglycerides through
cleavage of a chain of triglycerides. The increase of diacyglyceride
amount in the case of BHT indicates the cleavage of the acyl group
from triacylglyceride.
The presence of oxidized triacylglycerol species are indirectly
indicated in the base peak chromatogram in the presence of BHT
(see Fig. 4). In general, chromatographic peaks with higher
masses appeared later in the base peak chromatogram.
However, in the case of the peaks designated as (g), (h), and
(i) showed higher masses than those of other peaks that
appeared in 33.47–39.38 min (Fig. 3 and designated as (j), (k),
(l), (m) in Fig. 4), which indicates the different compositions of
triacylglycerols in the oxidized soybean oil. The unambiguous
identifications of these oxidized species could be made by MS/
MS experiments. For example, the chromatographic peak (h) in
Fig. 4 was subjected to MS/MS, and its resulting product mass
spectrum is shown in Fig. 6. Interestingly, a series of fragment
peaks were observed at m/z 599.5, 601.4, 615.4, and 617.4. These
peaks are tentatively assigned to arise from an oxidized
triacylglycerol with 18:1/18:2/18:2 sn chains, OLL, as indicated
in the inset of Fig. 6. Depending on the position of the oxidation
site, a multitude of oxidized isomers can be formed. In our MS/
MS spectrum, the fragments that can be generated from those
isomers were clearly observed. Detected oxidized triacylglycer-
ols are summarized in Table 5 and designated as LLL + 16 (g),
LLO + 16 (h), and OOL + 16 (i).
The diacylglyceride oxides in the case of BHT were identified as
LL-oxide and designated as (a) and LO-oxide as (b) in Fig. 4. The
oxidation (epoxidation) of the double bond could be explained by
the reaction of hydrogen peroxide and/or hydroperoxides formed
542 K.-W. Lee et al. / Journal of Industrial and Engineering Chemistry 17 (2011) 537–542

Table 5
Composition of triacylglyceride oxides in SBO in the presence of each antioxidant.

TAG oxide M.W. NH4+ form TAG-VC (%) TAG-VE (%) TAG-N-oxyl (%) TAG BHT (%)

POP + 16 846 864 13.57 15.01 14.12 17.61

LnLP + 16 870 888 9.07 10.13 11.81 17.07
PLL + 16 872 890 0.00 1.89 7.99 6.68
LLLn + 16 892 910 15.94 8.88 6.30 5.29
LLL + 16 984 (g)912 25.31 25.05 17.16 19.24
LLO + 16 896 (h)914 22.72 22.03 21.22 21.55
OOL + 16 898 (i)916 9.97 12.48 14.66 9.24
OOO + 16 900 918 3.43 4.53 6.74 3.32

Total 100.00 100.00 100.00 100.00

[(Fig._6)TD$IG] Abbreviation: VC = vitamin C = ascorbic acid, VE = vitamin E = a-Tocopherol, BHT = 2,6-di-t-butyl-4-cresol, N-oxyl = 4-hydroxy-TEMPO.

Fig. 6. MS/MS spectrum of a peak of (h) in Fig. 4.

during the autoxidation of SBO. It could be easily decomposed by
the acidic antioxidant.
4. Conclusion
In the investigation of antioxidant effect of two naturals,
ascorbic acid and a-tocopherol and two synthetics, BHT and
4-hydroxy-TEMPO, the ascorbic acid showed the synergetic and
antagonistic effect according to the concentration while
4-hydroxy-TEMPO showed only the antagonistic effect. The
synergetic effect at lower concentration is attributable to
the preferred inhibiting ability through less inter- and/or intra-
hydrogen bonding tendency of ascorbic acid than in high
concentration.
The composition of each oxidized soybean oil in the presence of
different antioxidants was confirmed by means of LC–ESI-MS/MS
spectrometry.
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