ARTICLE IN PRESS

LWT 38 (2005) 193–200

Antioxidant activity of peanut seed testa and its antioxidative

component, ethyl protocatechuate
Wen-Jye Yen, Lee-Wen Chang, Pin-Der Duh*
Department of Food Science and Technology, Chia Nan University of Pharmacy and Science, 60 Erh-Jen Road, Section 1, Pao-An,
Jen-te Hsiang, Tainan Hsien, Taiwan, Republic of China
Received 24 March 2004; received in revised form 28 May 2004; accepted 7 June 2004

Abstract

The antioxidant activity of ethanolic extracts of peanut seed testa (EEPST) and its antioxidative component, ethyl
protocatechuate (EP), was examined. It was found that EEPST and EP showed a dose-dependent activity on the inhibition of
liposome peroxidation. EEPST and EP in the range of 50–500 mg/l were effective in protecting protein against oxidative damage.
EEPST and EP at 100 mg/l showed 92.6% and 84.6% scavenging effect, respectively, on a,a-diphenyl-b-picrylhydrazyl radical,
indicating that they act as a primary antioxidant. In addition, EEPST and EP, at a dose of 200 mg/l, showed 70.6% and 67.7%
scavenging effect, respectively, on the hydroxyl radical. EEPST also exhibited a metal-binding ability, while EP did not. The
inhibitory effect of EEPST on linoleic peroxidation correlated with their polyphenolic content. These results suggest that the
antioxidant mechanism, for both EEPST and EP, could possibly be due to their scavenging effect on free radical and hydroxyl
radical. In addition, its metal binding ability may contribute to antioxidant activity of EEPST.
r 2004 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

Keywords: Peanut seed testa; Ethyl protocatechuate; Antioxidant activity; Polyphenolic compound

1. Introduction dietary intake of antioxidative compounds and the assay

Reactive oxygen species (ROS) are various forms of
activated oxygen, which include free radicals such as
hydroxyl and superoxide as well as non-free radicals
such as hydrogen peroxide. ROS are known to influence
the pathogenesis of certain human diseases, and play an
important role in the deterioration of foods during
storage. Although human beings have antioxidant
defenses against oxidative damage, these antioxidants
in human beings can be inefficient. In addition, synthetic
antioxidants such as butylated hydroxytoluene (BHT)
and butylated hydroxyanisole (BHA) are commonly
used in processed foods. However, there has been
growing concern over their safety and toxicity (Ito
et al., 1986). Moreover, some studies (Lu & Foo, 2000)
have reported that there is an inverse relationship
between dietary intake of antioxidant-rich foods and
the incidence of human diseases. Therefore, research on
*Corresponding author. Tel.: +886-62667302; fax: +886-62668340.
E-mail address: ipdduh@mail.chna.edu.tw (P.-D. Duh).
of the natural antioxidant source has attracted much
attention.
Peanut is a principal agricultural plant in the world.
The antioxidative activity of peanut has been investi-
gated (Pratt & Miller,1984; Yen, Wang & Duh,1990).
Our previous research on antioxidant activity in peanut
hulls (Duh, Yeh &Yen, 1992; Duh &Yen, 1995; Yen &
Duh, 1996) demonstrated that marked antioxidant
activity and antimutagenic effect are found in peanut
hulls, and the antioxidative component was identified as
luteolin.
The byproducts of agro-industries such as seed testa,
hulls and peels are usually wasted. The extraction and
identification of antioxidant components from hulls
(Duh et al., 1992), coats (Muamza, Robert, & Sparks,
1998; Chang, Yen, Huang, & Duh, 2002), and peels
(Larrauri, Ruperez, & Saura-Calixto, 1998) have been
reported recently. Peanut comprises kernels, seed testa
and hulls. Previous studies have shown that peanut
kernels and peanut hulls contained antioxidant compo-
nents, and the antioxidant properties of peanut hulls

0023-6438/$30.00 r 2004 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2004.06.004
 ARTICLE IN PRESS
194 W.-J. Yen et al. / LWT 38 (2005) 193–200
and kernels had been investigated. In addition, in our
previous study, the peanut seed testa has been found to
show antioxidant activity, and ethyl protocatechuate
(EP) was isolated and identified from peanut seed testa
(Huang, Yen, Yen, Chang, & Duh, 2003). However, the
antioxidant property of EP and the mechanism of
antioxidant activity of peanut seed testa remain unclear.
In recent years, increasing interest has been directed
towards the utilization of normal food constituents with
antioxidative properties. Hence, investigation of anti-
oxidant activity of peanut seed testa and EP is needed.
The purpose of this paper was to investigate the
antioxidant properties of peanut seed testa and EP,
and to elucidate the mechanism of antioxidant activity
of peanut seed testa.
2. Material and methods
2.1. Material
Peanut seed testa obtained from Sank Kong Corpora-
tion (Yunlin, Taiwan, R.O.C.) were stored at 4 C until
used. Lionleic acid, ammonium thiocyanate, ferrous
chloride, and butuylated hydroxyanisole (BHA) where
purchased form E. Merck (Darmstadt, Germany). All
other reagents were of analytical grade.
2.2. Extraction
The peanut seed testa (5.0 g), Spanish type, were
extracted overnight with 200 ml of ethanol in a shaking
incubator at room temperature. The extracts were
filtered, and the extraction was repeated twice. The
combined filtrates were evaporated to dryness in
vacuo and weighed to determine the yield of soluble
constituents.
2.3. Antioxidant activity in a linoleic acid system
Antioxidant activity assay was carried out by using
the linoleic acid system. Linoleic acid emulsion
(0.02 mol/l) was prepared with linoleic acid (0.2804 g)
and Tween 20 (0.2804 g) in phosphate buffer (50 ml, pH
7.4). A reaction solution containing each extract (0.2 ml,
0.25–2.5 mg/ml), linoleic acid emulsion (2.5 ml), and
phosphate buffer (2.3 ml, 0.2 mol/l, pH 7.0) were mixed
with a homogenizer. The reaction mixture was incu-
bated at 37 C in dark, and the degree of oxidation was
measured according to the thiocyanate method (Misuda,
Yasumoto & Iwami, 1966), by sequentially adding
ethanol (4.7 ml, 75 ml/100 ml), ammonium thiocyanate
(0.1 ml, 30 g/100 ml), sample solution (0.1 ml) and
ferrous chloride (20 mmol/l in 3.5 ml/100 ml HCl)
solution (0.1 ml). After the mixture was stirred for
3 min, the peroxide value was determined by reading the
absorbance at 500 nm, and the inhibition percent of
linoleic acid peroxidation was calculated as (%)
inhibition=[1 (absorbance of sample at 500 nm)/
(absorbance of control at 500 nm)]  100. All tests were
run in duplicate, and analysis of all samples were run in
triplicate and averaged.
2.4. Determination of antioxidant action on liposome
oxidation
Lecithin (580 mg) was sonicated in an ultrasonic
cleaner (Branson 8210, Branson ultrasonic Corporation,
Danbury, CT, USA) in 58 ml, 10 mmol/l phosphate
buffer (pH 7.4) for 2 h. The sonicated solution (10 mg
lecithin/ml), FeCl3, ascorbic acid and extracts (0.2 ml,
0.25–2.5 mg/ml) were mixed to produce a final concen-
tration of 3.12 mmol/l FeCl3, and 125 mmol/ml ascorbic
acid. The mixture was incubated for 1 h at 37 C by the
thiobarbituric acid (TBA) method (Tamura & Shiba-
moto, 1991). The absorbance of the sample was read at
532 nm against a blank, which contained all reagents
except lecithin.
2.5. Determination of effect on hydroxyl radical
The determination was carried out as described by
Halliwell, Gutteridge and Aruoma (1987). The reaction
mixture (3.5 ml), which contained extracts (0.2 ml, 0.05–
50 mg/ml), deoxyribose (6 mmol/l), H2O2 (3 mmol/l),
KH2PO4–K2HPO4 buffer (20 mmol/l, pH 7.4), FeCl3
(400 mmol/l), ethylenediaminetetraacetic acid (EDTA;
400 nmol/l), and ascorbic acid (400 nmol/l) was incu-
bated at 37 C for 1 h. The extent of deoxyribose
degradation was tested by the TBA method. One
millilitre of 0.01 g/ml TBA and 1 ml of 0.028 g/ml
trichloroacetic acid (TCA) were added to the mixture,
which was then heated in a water-bath at 90 C for
20 min. The absorbance of the mixture was read
spectrophotometrically at 532 nm. All analyses were
run in three replicates and averaged.
2.6. Determination of effect on DPPH radical
The effect of extracts on DPPH radical was estimated
according to the method of Hatano, Kagawa, Yasahara,
and Okuda (1988). The extracts (0.2 ml, 0.25–2.5 mg/ml)
were added to a methanolic solution (1.0 ml) of DPPH
radical (final concentration of DPPH was 0.2 mmol/l).
The mixture was shaken vigorously and allowed to
stand at room temperature for 30 min; the absorbance of
the resulting solution was then measured spectrophoto-
metrically at 517 nm. The tests were run in duplicate,
and analyses of all samples were run in triplicate and
averaged.
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W.-J. Yen et al. / LWT 38 (2005) 193–200
2.7. Determination of effect on protein oxidation
The effects of EEPST on protein oxidation was
carried out according to the method of Lenz, Costabek,
Shaltiel, and Cevine (1989). The reaction mixture
(1.2 ml), containing sample (0.25–2.5 mg/ml), phosphate
buffer (20 nmol/l, pH 7.4), bovine serum albumin
(20 mg/ml), FeCl3(100 mmol/l), H2O2 (2.0 mmol/l), as-
corbic acid (200 mmol/l), was incubated for 1 h at 37 C,
and 1 ml 20 nmol/l dinitrophenyl hydrazine (DNPH) in
2 mol/l HCl was added to the reaction mixture. One ml
cold TCA (0.2 g/ml, g/ml) was added to the mixture and
centrifuged at 3000  g for 10 min. The protein was
washed three times with 2 ml ethanol/ethyl acetate(1:1,
ml/ml) and dissolved in 2 ml 6 mol/l guanidine-HCl (pH
6.5). The absorbance of the sample was read at 370 nm.
Triplicate sample were run for each set.
2.8. Measurement of chelating activity on metal ions
The chelating activity of sample on Fe2+ was
measured according to the method of Carter (1971).
Briefly, extracts (0.2 ml, 0.25–2.5 mg/ml) were incubated
with 0.05 ml of FeCl24H2O (2.0 mmol/l). The reaction
mixture was initiated by the addition of 0.2 ml ferrozine
(5.0 mmol/l), and finally quantified to 0.8 ml with
methanol. After the mixture had reached equilibrium
(10 min), the absorbance at 562 nm was read. EDTA
served as the positive control, and a sample without
EEPST, EP or EDTA served as the negative control.
Triplicate samples were run for each set and averaged.
2.9. Determination of total polyphenolic compounds
The concentration of phenolic compounds was
measured according to the method of Taga, Miller and
Pratt (1984) and calculated using gallic acid as standard.
A sample (0.1 ml) was added to 2.0 ml of 0.02 g/ml
Na2CO3. After 2 min 0.5 ml/ml Folin–Ciocalteu reagent
(100 ml) was added to the mixture, which was then left
for 30 min. Absorbance was measured at 750 nm using a
spectrophotometer.
3. Results and discussion
Phospholipids, named as derivatives of phosphatidic
acid such as phosphatidylcholine (lecithine), are gen-
erally useful as syngerists in reforcing the antioxidant
activity of phenolic compounds. However, the auto-
xidation of phospholipids is similar to that of fatty
esters. Phospholipid is generally thought to be mainly
responsible for the oxidative deterioration and off-flavor
development of foods, due to its greater degree of
unsaturation (Wu & Sheldon,1988). Hence, phospholi-
pid may be regarded as a more relevant substrate for
195
evaluating antioxidant activity in lipid food system. In
addition, phospholipids are believed to be present in
high amounts in cell membranes. In order to investigate
in detail EEPST and EP in biological systems, the
phospholipid, prepared as a liposome, was used as the
model system for evaluating the inhibitory effect of
EEPST and EP against lipid peroxidation in cell
membranes.
The inhibitory effect of EEPST and EP on TBARS
formation in a liposome model system is shown in
Fig. 1. EEPST and EP in the range of 50–500 mg/l
showed 28.0–85.8% and 26.2–29.1% inhibition of
TBARS formation, respectively, compared with the
control. The inhibition was concentration-dependent.
Of the four samples tested, the inhibitory effect of
EEPST was higher than that of EP, Toc and BHA. EP
was derived from purification of EEPST. EEPST
showed higher antioxidant activity than EP because
the combined action of substances makes the antiox-
idant activity of the crude extracts higher than that of an
isolated compound (EP). As shown in Fig. 1, EEPST
and EP reduce TBARS formation, indicating that
EEPST and EP may inhibit the oxidation of lipids in
food system as well as protect cell membranes against
oxidative damage.
The effect of EEPST and EP on protein carbonyl
formation in albumin, induced by FeCl3, H2O2 and
ascorbic acid is plotted in Fig. 2. EEPST and EP in the
range of 50–500 mg/l show 2.2–97.2% and 0.3–96.3%
inhibition of protein oxidation, respectively. As shown
in Fig. 2, the protective effect of EEPST and EP are
Fig. 1. Antioxidant activity of ethanolic extracts of peanut seed testa
(EEPST, ) and ethyl protocatechuate (EP, J) in a liposome model
system. Toc (tocopherol; ’); BHA (butylated hydroxylanisole, &).
Values represent mean 7SD (n=3).
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196 W.-J. Yen et al. / LWT 38 (2005) 193–200

confirms that EEPST and EP have the ability to

enhance the stability against primary oxidation. In
other words, EEPST and EP are free radical inhibitors,
as well as primary antioxidants that react with free
radicals.
Fig. 4 shows the chelating effect of EEPST on ferrous
ions. The chelating effect increased with an increased

Fig. 2. The protective effect of ethanolic extracts of peanut seed testa

(EEPST, ) and ethyl protocatechuate (EP, J) on protein oxidative
damage. Toc, (tocopherol; ’); BHA(butylated hydroxylanisole, &).
Values represent mean 7SD (n=3).

significantly more remarkable than that of Toc and

BHA. Stadman (1992) noted that some amino acid
residues are oxidized to carbonyl derivatives; conse-
quently, it becomes evident that carbonyl amounts of Fig. 3. The scavenging effect of ethanolic extracts of peanut seed testa
proteins undergoing oxidation could be used as a (EEPST, ) and ethyl protocatechuate (EP, J) on DPPH radicals.
Toc (tocopherol; ’); BHA (butylated hydroxylanisole, &). Values
measure of protein damage, even though measures of
represent mean 7SD (n=3).
protein carbonyls are not specific and are influenced by
factors other than peroxidation. Teissedre , Frankel,
Waterhouse, Peleg and German (1996) reported that
polyphenolics can bind with protein surfaces. As seen in
Fig. 2, the fact that the oxidative damage of bovine
serum albumin was inhibited by EEPST and EP may be
concerned with binding with bovine serum albumin.
It is well-known that free radicals induce lipid
peroxidation, proceeding like that of many other
organic compounds by a free radical chain mechanism.
To elucidate the relationship between free radicals and
the extracts, a,a-diphenyl-2-picrylhydrazyl (DPPH) was
used as the substrate in the present work. The
scavenging effect of EEPST and EP on the DPPH
radical is shown in Fig. 3. The decrease in absorbance of
the DPPH radical caused by antioxidant was due to the
scavenging of the radical by hydrogen donation. It is
visually noticeable as a discoloration from purple to
yellow color. The scavenging effect of EEPST and EP in
the range of 50–500 mg/l on the DPPH radical increased
with an increasing concentration of EEPST and EP.
EEPST and EP at 100 mg/l showed 92.6% and 84.6%
Fig. 4. The chelating effect of ethanolic extracts of peanut seed testa
scavenging effect on DPPH radical, respectively, in- (EEPST, ) and ethyl protocatechuate (EP, J) on iron ion. EDTA
dicating that EEPST and EP were sufficient to cause a (ethylenediaminetetraacetic acid, ’). Values represent mean 7SD
noticeable effect on scavenging free radicals. This (n=3).
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W.-J. Yen et al. / LWT 38 (2005) 193–200
concentrations of EEPST. EEPST at 200 mg/l showed
63.4% chelating activity on ferrous ion. Unlike EEPST,
there was no chelating effect in EP at the concentration
(50–100 mg/l). Iron is essential for life because it is
required for oxygen transport, respiration and the
activity of many enzymes. However, iron is an extremely
reactive metal and will catalyze oxidative changes in
lipid, protein, and other cellular components (Decker &
Hultin, 1992). In addition, the liposome peroxidation,
and oxidative damage of protein model systems were
induced by a Fenton reaction in which ferrous ions
catalyze the composition of hydrogen peroxide to
hydroxyl anion and hydroxyl radical with the produc-
tion of ferric ion. As shown in Fig. 4, the cheating effect
of EEPST on ferrous ions is relatively lower when
compared with that of EDTA, it may be significant
because EEPST minimises the concentration of metal in
the Fenton reaction.
The
OH-scavenging effects of EEPST and EP were
investigated at the concentration of 0–500 mg/l. Fig. 5
shows the
OH-scavenging effects by the 2-deoxyribose
oxidation method. The scavenging effect of EEPST and
EP on hydroxyl radical was concentration-dependent.
EEPST and EP at 200 mg/l exhibited 70.6% and 67.7%
scavenging effect on hydroxyl radical, respectively,
which were significantly higher than that of Toc
(44.3%) and BHA (42.4%). In the model system, the
mixture of FeCl3–EDTA and H2O2 is incubated with
deoxyribose in phosphate buffer (pH 7.4), the hydroxyl
radicals generated attack the deoxyribose and result in a
series of reaction forming TBARS (Halliwell, Gutter-
Fig. 5. Hydroxyl radical scavenging effects of ethanolic extracts of
peanut seed testa (EEPST, ) and ethyl protocatechuate (EP, J) by
the 2-deoxyribose oxidation method. Toc (tocopherol, ’); BHA
(butylhydroxyl anisole, &). Values represent mean 7SD (n=3).
197
idge, & Aruoma, 1987). Consequently, the ability to
diminish the color formation has been adapted not only
as a measurement of antioxidant properties (Aruoma,
1991) but also as an assay of protective deoxyribose
from oxidative damage. Zhao and Jung (1995) reported
that any hydroxyl radical scavenger added to the
reaction mixture would compete with deoxyribose for
hydroxyl radicals to an extent depending on its rate
constant for reaction with hydroxyl radicals and its
concentration relative to deoxyribose. According to
results obtained, the rate of constant of EEPST and EP
for reaction with hydroxyl radicals was greater than that
of deoxyrobose; therefore EEPST and EP can protect
deoxyribose from oxidative damage. Among the reactive
oxygen species, hydroxyl radical is the most active.
Indeed, it is one of the most reactive chemical species
known. The hydroxyl radical induces some oxidative
damage to biomolecules such as all proteins, DNA,
PUFA, nucleic acid, and almost any biological molecule
it touches, and this damage causes aging, cancer and
several diseases (Aruoma, 1998). According to the
present findings, EEPST and EP might be used to
provide a good hydroxyl radical scavenger for humans
and foods.
Previous reports noted that polyphenolics are asso-
ciated with antioxidant activity and play an important
role in stabilizing lipid peroxidantion (Yen, Duh, &
Tsai,1993). The equation of the amounts of polyphe-
nolics (y) and the amount of EEPST (x) used is
y=0.1033950921x+0.0105774105 (r=0.87, Po0.05),
indicating that the amounts of polyphenolics increased
with an increased amount of EEPST. In other words,
each one mg of EEPST contains 0.0315 mg of
polyphenlolics. The antioxidant activity and the
amounts of phenolics in EEPST is shown in Fig. 6.
The equation of the antioxidant activity (y) and
the amounts of polyphenylics (x) in EEPST
used is y=792.0519878724x+28.3832011559 (r=0.71,
Po0.05), revealing that the correlation between anti-
oxidant activity of EEPST in a linoleic acid model
system and the amounts of polyphenolics was signifi-
cant. Phenolic compounds could easily donate a
hydroxy hydrogen due to resonance stabilization
(Fessenden & Fessenden, 1994). In addition, EEPST
showing high radical scavenging potential (Figs. 3 and
5) might be explained by their ability to donate a
hydrogen atom from their phenolic hydroxy groups
(Sawa, Natao, Akaike, Ono, & Maeda, 1999). Combin-
ing this fact with the results obtained, we suggest that
polyphenolics in EEPST may contribute directly to
antioxidant activity of the extracts. Diet appears to play
important role in human health and in the development
of certain disease (Ames, 1983). Considerable attention
has been focused on food phytochemicals that occur
naturally or are added to food and beverages for human
consumption (Wattenberg, 1978; Slaga, 1981). It is
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198 W.-J. Yen et al. / LWT 38 (2005) 193–200
Fig. 6. The correlation between total phenolic content and antioxidant
activity of ethanolic extracts of peanut seed testa (EEPST, ). The
antioxidant activity was determined according to the thiocyanate
method. Values represent mean 7SD (n=3).
suggested that some polyphenolics have inhibitory
effects on mutagenesis and carcinogenesis in human,
when up to 1.0 g daily ingested from diet rich in fruits
and vegetables (Tanaka, Kuei, Nagashima, & Taguchi,
1988). Our results show that EEPST contained a higher
amount of polyphenolics. This suggests that when
consuming peanuts, the peanut seed testa should not
be removed due to its high content of polyphenolic
compounds. This finding may be significantly beneficial
and meaningful to consumers.
The mechanism of aging of plants in living system is
believed to involve reactions related to lipid and non-
lipid peroxidation. Certain inherent defense system
governs such reaction and facilitates the maintenance
of vital physiological processes; the defense system must
be located in the coat and in the hulls (Ramarathnam,
Osawam, Namiki, & Kawakishi, 1989). In previous
studies (Yen et al., 1993), high levels of phenolic
compounds (0.167 mg/g of hull) were found to exist in
peanut hulls and an antioxidant component was
identified as luteolin. In addition, EP, an antioxidative
component, was purified and identified from peanut
seed testa (Huang et al., 2003). Regarding the biological
effects of EP, it has so far been well elucidated. Yen and
Hsieh (2002) noted that protocatechuic acid was a major
source of the inhibitory effect against LDL oxidative
modification induced by Cu2+. Recently, protocate-
chuic acid has been demonstrated to be an efficacious
agent in different tissues, such as diethylnitrosamine in
the liver (Tanaka, Kojima, Kawamori, Yoshimi & Mori,
1993), 4-nitroquinoline-1-oxide in the oral cavity (Ta-
naka et al., 1994), azoxymethane in the colon (Kawa-
mori et al., 1994), N-methyl-N-nitrosourea in glandular
stomach tissue (Tanaka, Kojima, Kawamori & Mori,
1995), and N-butyl-N-(4-hydroxybutyl) nitrosamine in
the bladder (Hirose, Tanaka, Kaeamori, Ohnishi, Mori,
1995). Apparently, EP shows a potent pharmacological
and biological activity on certain disease. Although data
regarding the biological effects except antioxidant
activity of EP are limited in the present study, our
previous study (Huang et al., 2003) showed that EP play
an important role in preventing lipid oxidation, and
contributed to the antioxidant activity of EEPST.
Moreover, EP may in part be responsible for the defense
system of peanut seed testa. In addition, EP may be
utilised in food preservation and protect lipid against
oxidation, however, overdoses of EP (500 mg/kg) has a
potential to enhance tumorigenesis, induce contact
hypersensitivity in mouse skin and can disturb the
detoxification of ultimate carcinogen (Nakamura,
Torikai, & Ohigashi, 2001). Hence, if EP is used as
food additives or phytochemicals, the safety and
toxicology of EP have to be distinguished in detail and
extensively studied.
Kaneko, Baba and Matsuo (2000) noted that the
protection by phenolic antioxidant against the cytotoxi-
city of phosphatidylcholine hydroperoxides was
presumed to depend on their affinity for phosphatidyl-
cholines and on their orientation in the membranes. The
data presented here demonstrated that EEPST and EP
protected lipids and non-lipids from oxidative damage.
There is a direct correlation between inhibitory oxida-
tive damage and phenolic compounds. In addition, the
protective effect of EEPST is well associated with the
amounts of polyphenolic compounds (Fig. 6). This is
because the polyphenolics in EEPST exert their redox
properties, allowing them to act as hydrogen donators
(Fig. 3), and as hydroxyl radical quenchers (Fig. 5). In
addition, they have a metal chelating ability (Fig. 4) and
a protein binding potential (Fig. 2). In other words, the
antioxidant activity of phenolic compounds in EEPST is
related not only to structure characteristics, but also to
their ability to interact with the interface where
oxidation takes place (Heinonen et al., 1998). Further
studies on the antioxidant activity of EEPST and EP
in vivo are in progress.
Acknowledgements
The assistance of Mr. Jui Lin Li is gratefully
acknowledged. This research work was supported by
the National Science Council, Republic of China, under
Grant NSC91-2313-B-041-002.
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