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Antioxidant Activity of Durian (Durio zibethinus Murr.) Shell in vitro

Article  in  Asian Journal of Pharmaceutical and Biological Research · January 2011

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 Asianand
maceutical Journal of Pharmaceutical
Biological Research and Biological
Original Research
Research Short Communication

Antioxidant Activity of Durian (Durio zibethinus Murr.) Shell

In vitro

Li Wang and Xican Li*

School of Chinese Herbal Medicine, Guangzhou University of Chinese Medicine, Guangzhou,
China E mail :lixican@126.com

Publication History
Received :22 Oct 2011
Accepted :28 Nov 2011
Published :10 Dec 2011
Key Words
Durian shell
Antioxidant activity
Durio zibethinus Murr
Metal chelating
Radical-scavenging
Total phenol
Abstract
Objective: The paper tried to systematically investigate the antioxidant potential of
Durian (Durio zibethinus Murr.) shell in vitro then to provide the evidence for its
further development in future.
Methods: The antioxidant activity of methanol extract of durian shell (MEDS) was
evaluated using 1,1-diphenyl-2-picryl-hydrazyl (DPPH·) radical, 2,2'-azino-bis (3-
ethylbenzthiazoline-6-sulfonic acid) (ABTS+·) radical, reducing power (Fe3+ & Cu2+),
superoxide anion radical (O·2- ), hydroxyl radical (·OH), anti-lipid peroxidation and
ferrous ions (Fe2+) chelating activity assays, compared with positive control Trolox.
The total phenol contents were also determined by Folin-Ciocalteu reagent.
Results: MEDS exhibited effective antioxidant abilities in dose-dependent manners,
its IC50 values were calculated as 102.37±1.98, 19.50±1.44, 280.79±22.80,
154.67±5.84, 770.52±19.28, 324.63±17.61, 4.45±1.13 and 63.95±14.91 ìg/mL,
respectively, for DPPH, ABTS+, reducing power (Fe3+), reducing power (Cu2+),
superoxide anion scavenging, hydroxyl scavenging, anti-lipid peroxidation, chelating
ability (Fe2+) assays. The total phenol contents (33.77±1.77 mg GAE/g) were found in
MEDS.
Conclusion: Durian (Durio zibethinus Murr.) shell possesses effective in vitro
antioxidant ability which may contribute to its pharmacological effects or healthcare
functions. It exerted the antioxidant possibly via metal-chelating and radical-
scavenging which may result from donating hydrogen atom (H·) and electron (e). Its
antioxidant ability can be attributed to the total phenol. As a waste generally discarded,
durian shell can be a source of useful natural antioxidant for medical or health care.

INTRODUCTION that these effects may be associated with its

D urian (Durio zibethinus Murr) is a
typical and very popular tropical
fruit, which is commonly consumed around the
world. Its fruit was proved to be of antimicrobial
potential [1], but the shell is generally discarded as a
waste.
Until now, the utility of durian shell is still
very limited. It was reported that durian shell can be
used to prepare activated carbon adsorbent for
[2] [3]
chemical materials science . In traditional
Chinese medicine (TCM), it can benefit qi,
replenish blood and nourish yin. In fact, it is often
stewed with chicken to prepare a health-care soup
in Guangdong Province of China. In addition, its
extract was proved to have protective effect on
stress-induced liver injury in mice [4]. According to
[5]
free radical biology and medicine , we supposed
antioxidant. However, there is no report concerning
its antioxidant activity yet.
Therefore, the paper tried to systematically
investigate its antioxidant effects by various assays
then provide the evidence for its further
development in future.
Materials and Methods
Chemicals
Trolox(±-6-hydroxyl-2,5,7,8-
tetramethlychromane-2-carboxylicacid),DPPH
(1,1-Diphenyl-2-picrylhydrazyl radical),
pyrogallol, Ferrozine [3-(2-pyridyl)-5,6-bis (4-
phenylsulfonicacid)-1,2,4-triazine], neocuproine
(2,9-dimethyl-1,10-phenanthroline), linoleic acid,
were purchased from Sigma Co. (Sigma-Aldrich
S h a n g h a i Tr a d i n g C o . , C h i n a ) ; A B T S

e ISSN: 2231-2218 542

© 2011 Asian J Pharm Biol Res

diammonium salt [2,2'-Azino-bis (3-ethyl
benzothiazoline-6-sulfonic acid diammonium
salt)], and D-2-deoxyribose were obtained from
Amresco Co.; All other chemicals used were in
analytical grade.
Preparation of methanol extract of durian shell
(MEDS)
Durian was obtained from Jialianfu
Supermarket, Changzhou Island, Guangzhou,
China. The fresh shell was cut into pieces, dried at
room temperature then powdered. The powder was
extracted exhaustively in a Soxhlet extractor with
methanol. The extract was then concentrated in
vacuo until the solvent was completely removed.
The methanol extract of durian shell (MEDS)
obtained was kept in a well-closed container in
refrigerator until use.
DPPH· scavenging activity
DPPH· radical-scavenging activity of
MEDS was measured by the method as described
by Li [6]. Briefly, 1 mL of DPPH· solution (0.1 mM)
was mixed with 0.5 mL of various concentration
(20, 40, 60, 80, 100 ìg/mL) of samples dissolved in
95% methanol. The mixture was kept at room
temperature for 30 min, and then the absorbance at
519 nm was measured on a spectrophotometer
(Unico 2100, Shanghai, China), using 95%
methanol as the blank. BHA, Trolox were used as
the positive controls, and the DPPH· inhibition
percentage was calculated:
Inhibition % = (1 – As/A0) × 100 %
Where As is the absorbance in the presence of
samples, while A0 is the absorbance without
samples.
+ presence of samples, while A0 is the absorbance
ABTS· scavenging activity
The scavenging activity of ABTS·+ was
determined as described by Li [6]. The ABTS·+ was
produced by the reaction between 0.35 mL of
ABTS diammonium salt (7.4 mM) and 0.35 mL of
potassium persulfate (2.6 mM), stored in the dark at
room temperature for 12 hours to allow completion
of radical generation. Before usage, the mixture
was diluted with 95% methanol (about 1:50) to get
an absorbance of 0.70±0.02 at 734 nm on a
spectrophotometer (Unico 2100, Shanghai, China).
To determine the scavenging activity, 1.2 mL of
ABTS·+ reagent was mixed with sample( 5, 10, 20,
40, 60 ìL), the total volume of system was adjusted
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Asian J Pharm Biol Res Oct-Dec 2011 Vol. 1(4)
to 1500 ìL with methanol. The absorbance at 734
nm was measured 6 min after the initial mixing,
using 95% methanol as the blank. The percentage
inhibition of the samples was calculated as:
Inhibition % = (1 – As/A0) ×100 %
Where As is the absorbance in the presence of
samples, while A0 is the absorbance without
samples.
Ferric ions (Fe3+) reducing power
3+
Ferric cyanide (Fe ) reducing power was
[7]
determined by the method of Oyaizu . In brief, x
ìL samples (x =0, 30, 75, 130, 175, 210 ìL) were
mixed with Na2HPO4/KH2PO4 buffer (350-x ìL, 0.2
M, pH 6.6) and K3Fe(CN)6 (250 ìL, 1 g/100 mL).
The mixture was incubated at 50 ºC for 20 min, 250
ìL of TCA (10 g/100 mL) was added, and the
mixture was centrifuged at 3500 r/min for 10 min.
The upper layer (400 ìL) was mixed with distilled
water (400 ìL) and FeCl3 (400ìL, 0.1 g/100 mL)
and placed immediately into the spectrophotometer
(Unico 2100, Shanghai, China), and the timer was
started. The absorbance at 700 nm was measured at
90s. Samples were analyzed in groups of three, and
when the analysis of one group has finished, the
next group of three samples were mixed with FeCl3
to avoid oxidization by air. Trolox, BHA was used
as the positive control, and an increased absorbance
indicated increased reducing power. The
percentage reducing power of the sample as
compared to the maximum absorbance tested
which appeared in Trolox at 175 ìg/mL was
calculated by using the formula: (As-A0)/(Am –A0) ×
100. Here, A m = absorbance of maximum
absorbance tested, As is the absorbance in the
without samples.
2+
Cupric ions (Cu ) reducing power
The cupric ions (Cu2+) reducing power
capacity was measured by the method [8] with slight
modification. Briefly, 125 ìL CuSO4 aqueous
solution (10 mM), 125 ìL neocuproine ethanolic
solution (7.5 mM) and 500 ìL CH3COONH4 buffer
solution (100 mM, pH 7.5) were added to test tubes
with different volumes of sample (30, 60, 90, 120,
150 ìL). Then, the total volume was adjusted to 1
mL with the buffer then mixed vigorously.
Absorbance against a buffer blank was measured at
450 nm after 30 min. Increased absorbance of the

reaction mixture indicated an increase of reduction
capability. Trolox and BHA were used as the
positive controls. The percentage reducing power
of the sample as compared to the maximum
absorbance tested which appeared in sample at 150
ìg/mL was calculated using the formula: (As-
A0)/(Am–A0) × 100. Here, Am = absorbance of
maximum tested, As is the absorbance in the
presence of samples, while A0 is the absorbance
without samples.
- Where A0 is the absorbance at 532 nm of the
Superoxide anion (O2· ) radical-scavenging
activity
Measurement of superoxide anion
scavenging activity of sample was based on
Marklund method [9]. Briefly, samples were
dissolved in methanol at 1 mg/mL. The sample
solution (x = 50, 100, 150, 200, 250 ìL) was mixed
with Tris-HCl buffer (2920 - x ìL, 0.05 M, pH 8.2)
containing EDTA (1 mM) and pyrogallol (80 ìL, 6
mM), then shaken rapidly at room temperature. The
absorbance at 325 nm of the mixture was measured
(Unico 2100, Shanghai, China) against the Tris-
HCl buffer as blank every 30 s for 5 min. Trolox and
BHA were used as the positive controls. The slope
of the correlation of absorbance with time was
calculated. The reaction mixture without sample
was used as the negative control. The O2·–
scavenging ability was calculated as:
（１–
Slope of sample/Slope of negative
control) × 100 %
Hydroxyl radical (·OH) scavenging assay
The scavenging activity on the hydroxyl
radical (·OH) was investigated by the deoxyribose
method [10] modified with some modification. Our
preliminary experiments demonstrated that most
organic solvents can considerably promote the
inhibition percentage value. Hence, the
determination was performed as the following
procedure: all test samples (20, 50, 100, 200, 300,
400 ìL) were firstly dissolved with methanol in
mini tubes, the methanol solvents were then
o
completely removed at 80 C to eliminate the
interference. The reactions were performed in 0.2
M phosphate buffer (pH 7.4), containing 2.8 mM
deoxyribose, 2.8 mM H2O2, 25 ìM FeCl3, 80 ìM
Na2EDTA, and the test sample (20-400 ìg). The
reaction was started by adding ascorbic acid to a
final concentration of 100 ìM and the reaction
o
mixture (600 ìL in total) was incubated at 50 C in a
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Asian J Pharm Biol Res Oct-Dec 2011 Vol. 1(4)
water bath for 20 min. After incubation, the color
was developed by addition of 0.5 mL 2-
thiobarbituric acid (1 g/100 mL) followed by 0.5
mL trichloroacetic acid (5 g/100 mL) and heated in
a blast thermostated oven at 105 oC for 15 min with a
tight cover. The mixture was cooled then measured
its A532 nm against buffer (as blank). The scavenging
activity on hydroxyl radicals was expressed as:
Inhibition % = (1- A/A0) × 100 %
reaction mixture without sample, and A is the
absorbance at 532 nm of the reaction mixture
containing sample. Trolox and BHA were positive
controls.
Lipid peroxidation
The effect of sample on the prevention of
peroxidation in linoleic acid emulsion was
[11]
investigated using the thiocyanate method with
some modifications. The linoleic acid emulsion
was prepared by mixing and homogenizing 312.6
mg of linoleic acid, 78.2 mg of Tween-20 as
emulsifier, and 30 mL of 30% methanol (v/v), 0.1
mL of various concentrations of samples (3.75, 7.5,
11.25, 15, 18.75 ìg/mL) were added to 1.5 mL of
linoleic acid emulsion and 0.4 mL distilled water.
The reaction mixtures (2 mL) were incubated at
room temperature in open glass bottles. The degree
of oxidation was measured when the absorbance of
the control reached its maximum. To 0.15 mL
sample solution, 3.65 mL methanol, 0.1 mL
ammonium thiocyanate (30%, m/v), and 0.1 mL
ferrous chloride (0.02 mol/L in 3.6% HCl) were
added. The mixture was diluted two-fold with
methanol, then measured the absorbance at 500 nm
in a spectrophotometer (Unico 2100, Shanghai,
China). The mixture without sample was used as
control. The percentage of inhibition on lipid-
peroxidation was calculated by following equation:
Inhibition % = (1 – As/A0) × 100 %
Where As is the absorbance in the presence of
samples, while A0 is the absorbance without
samples.
2+
Chelating activity on Fe
The chelating activities on Fe2+ of MEDS
and positive controls were estimated by the method
[11]
. Briefly, the sample (20, 40, 60, 80,100 ìL) was
added to a solution of 250 ìM FeCl2 (100 ìL). The
reaction was initiated by the addition of 500 ìM

ferrozine (150 ìL) and total volume was adjusted to
1000 ìL with methanol. Then, the mixture was
shaken vigorously and left at room temperature for
5 min. Absorbance of the solution was then
measured on a spectrophotometer (Unico 2100,
Shanghai, China) at 562 nm. The percentage of
chelating effect was calculated by using the
formula given bellow:
Ferrous chelating effect (%) = (1- As/A0) × 100 %
Where As is the absorbance in the presence of
samples, while A0 is the absorbance without
samples.
Total phenol content
The total phenol content in MEDS was
estimated by the procedure [12] with some
modifications. Briefly, 500 ìL extract solution was
mixed with 500 ìL 0.25 M Folin-Ciocalteu reagent.
After 3 min, 1000 ìL of 15% Na2CO3 (W/V) was
added. The plastic tube was properly centrifuged at
3500 r/min for 3 min. A blue color was developed
for 30 min then the absorbance was read at 750 nm
using a spectrophotometer (Unico 2100, Shanghai,
China). Quantitative measurements were
performed based on a six-point standard calibration
curve of 10, 20, 40, 80, 120 mg/L of gallic acid in
methanol. The total phenol content was calculated
as a gallic acid equivalent (GAE) from the
calibration curve, and expressed as mg gallic acid
equivalent/g MEDS (mg GAE/g MEDS).
Statistical Analysis
The experiment was performed in
triplicate and the data were recorded as mean ± SD.
The IC 50 value was defined as the final
concentration of 50% radical inhibition. A value of
P < 0.05 was considered to be significant. The
statistical analysis was performed by SPSS (SPSS
Inc., Chicago, IL).
RESULTS AND DISCUSSION
Antioxidants are closely related to some
bio-functionalities, such as the reduction of chronic
diseases like DNA damage, mutagenesis,
carcinogenesis and inhibition of pathogenic
bacteria growth which is often associated with the
termination of free radical propagation in
biological systems [13]. Thus, antioxidant capacity is
widely used as a parameter for medicinal plant or
functional food.
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Asian J Pharm Biol Res Oct-Dec 2011 Vol. 1(4)
+
DPPH· and ABTS· scavenging activities
As DPPH· is a stable free radicals, it is
therefore widely used for radical scavenging assay
[8]
. The radical can be dissolved in methanol or
ethanol, and the color shows characteristic
absorptions at 519 nm. When an antioxidant
scavenges the free radical by donating hydrogen
(H·), the color in the DPPH· assay solution
becomes lighter. As presented in Figure1, the
sample and positive controls showed a
concentration-dependent manner within 20-100
ìg/ml. Hence, MEDS possessed effective
antioxidant ability on DPPH·. Their IC50 values
were listed in Table 1.
Unlike the reaction with DPPH· radical,
which involve H-atom transfer, the reaction with
ABTS+ radical involves an electron (e) transfer
process. The reduction capability of ABTS+ radical
was determined by the decrease in its absorbance at
734 nm induced by antioxidants. As seen in Figure
2, for the positive controls and sample, the ABTS+
inhibition percentage values were dose-dependent
in the range of 6.67-33.33 ìg/mL. Comparing to the
IC50 value of Trolox (1.45±0.11 ìg/mL), MEDS
(19.50±1.44 ìg/mL) was proved to possess an
effective antioxidant (Table 1).
Base on the fact that MEDS can effectively
inhibit both DPPH· and ABTS+·, it suggested that
MEDS exerted radical-scavenging action possibly
by donating hydrogen atom (H·) and electron (e).
Reducing power assay
3+ 2+
As seen in Figure 3 & 4, the Fe & Cu
reductive activities of sample and positive controls
were concentration dependent. There is a direct
correlation between antioxidant activity and
reducing power of an antioxidant [14], so reducing
capacity may serve as a significant indicator of its
[15]
potential antioxidant activity . The IC50 values in
Table 1 indicated that MEDS exhibited modest
antioxidant.
-
Superoxide anion (O·2 ) scavenging activity
Superoxide anion (O2· -) is one of the most
important free radicals in living cells. In the study,
the superoxide radical was generated by the
pyrogallol system at pH 8.2. Figure 5 showed that
positive controls and MEDS demonstrated an
ability to inhibit superoxide anion in dose-
dependent manners. As its IC50 value significantly
 Asian J Pharm Biol Res Oct-Dec 2011 Vol. 1(4)

Table 1 The values of IC50 of methanol extract of durian shell (MEDS) and positive controls
ìg/mL)

Assays MEDS Trolox BHA

Reducing power (Fe3+) 280.79±22.80 ab 43.84±7.48 45.90±7.48

Reducing power (Cu2+) 154.67±5.84 ab 12.28±0.99 7.42±0.17

• OH 324.63±17.61 ab 145.36±3.34 155.30±39.26

O2•- 770.52±19.28 ab 3385.45±1230.08 194.21±2.31*

Anti-lipid peroxidation 4.45±1.13 ab 0.31±0.01 0.45±0.01

DPPH• 102.37±1.98 ab 5.15±0.11 5.36±0.08

ABTS+• 19.50±1.44 ab 1.45±0.11 0.69±0.39

Chelating power 63.95±14.91 ab 392.90±39.33 7.70±0.45**

＊　 The positive control was GSH, instead of BHA. ** The positive control was sodium citrate, instead of BHA.
All values were mean ± SD (n=3). Results were analyzed by Independent-Samples T TestValue with a significantly
different from Trolox (P < 0.01); Value with b significantly different from BHA or GSH, sodium citrate (P < 0.01).

Figure 1. The radical-scavenging activity on DPPH of MEDS and positive controls

Each value is expressed as mean ± standard deviation, n =3.

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 Asian J Pharm Biol Res Oct-Dec 2011 Vol. 1(4)

Figure 2. The radical-scavenging activity on ABTS of MEDS and positive controls

Each value is expressed as mean ± standard deviation, n =3.

Figure 3. The Fe3+ reducing power of MEDS and positive controls

Each value is expressed as mean ± standard deviation, n =3.

Hydroxyl (·OH) scavenging activity
- deoxyribose degradation assay. A mixture of Fe -
Like superoxide anion (O2· ), hydroxyl
radical (·OH) is also regarded as one of the most
important ROS (reactive oxygen species ) in living
cells. Furthermore, it has a high reactivity with a
-9 [16]
very half-life of approx. 10 s in vivo .
In the study, the hydroxyl (·OH) radical
scavenging activity was evaluated using the
3+
EDTA, hydrogen peroxide (H2O2), and ascorbic
acid was used to generate hydroxyl radical
(·OH).The response degrades D-2-deoxyribose into
fragments which heated with thiobarbituric acid at
acidic solution, are detected at 540 nm because they
generate a pink chromogen.

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 Asian J Pharm Biol Res Oct-Dec 2011 Vol. 1(4)

Figure 4. The Cu2+ reducing power of MEDS and positive controls

Each value is expressed as mean ± standard deviation, n =3.

Figure 5. The O2·- radical-scavenging activity of MEDS and positive controls

Each value is expressed as mean ± standard deviation, n =3.

It can be observed that the inhibition Lipid peroxidation

percentages on ·OH of sample and positive controls
increased dose-dependently with the concentration
(Figure 6). Comparing to the positive controls
(Table 1), MEDS was proved to be an effective
antioxidant on ·OH.
Lipid peroxidation consists of a series of
free radical-mediated chain reaction processes and
is associated with several types of biological
damage.

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 Asian J Pharm Biol Res Oct-Dec 2011 Vol. 1(4)

Figure 6. The ·OH radical-scavenging activity of MEDS and positive controls

Each value is expressed as mean ± standard deviation, n =3.

Figure 7. The anti-lipid peroxidantion activity of MEDS and positive controls

Each value is expressed as mean ± standard deviation, n =3.

It can be seen in Figure 7 that the effects of sample
and positive controls on lipid peroxidation
increased in a concentration-dependent manner in
linoleic acid emulsion system. All IC50 values were
calculated and listed in Table 1.
2+
Fe chelating activity
As ferrous ion can catalyzed the oxidation
in body, it is important to investigate the metal
chelating activity of an antioxidant. As shown in
549
Figure 8, the Fe2+ chelating activities of positive
controls and MEDS increased dose-dependently
within 5-100 ìg/mL.
The IC50 values listed in Table 1 suggested
MEDS (63.95±14.91ìg/mL) had the stronger metal
chelating ability than Trolox
(392.90±39.33ìg/mL). It was therefore supposed
that MEDS exerted its antioxidant possibly via
radical-scavenging and metal-chelating.

Figure 8. The chelating ability on Fe2+ of MEDS and positive controls
Each value is expressed as mean ± standard deviation, n =3.
Total phenol Contents
The phenolics compounds in plant are
believed to be responsible for its antioxidant. Our
data revealed that there was high level of total
phenol content (33.77±1.77 mg GAE/g) in MEDS.
In the test, the equation of linear regression
between the concentration (mg/mL) of gallic acid
and the absorbance was calculated as: y = 152.24 x
+ 0.0683, R = 0.9962.
CONCLUSION
In conclusion, Durian (Durio zibethinus
Murr.) shell possesses effective in vitro antioxidant
ability which may contribute to its pharmacological
effects or healthcare functions. It exerted the
antioxidant possibly via metal-chelating and
radical-scavenging which may result from
donating hydrogen atom (H·) and electron (e). Its
antioxidant ability can be attributed to the total
phenol. As a waste generally discarded, durian shell
can be a source of useful natural antioxidant for
medical or health care.
REFERENCES
1. Dhasarathan P, Paulsi S: Evaluation of
Bioactive Potential in Durian Fruit (Durio
zibethinus) Samples Using Pathogens. Asian
Journal of Pharmaceutical and Biological
Research, 2011, 1: 1-7.
550
Asian J Pharm Biol Res Oct-Dec 2011 Vol. 1(4)
2. Thio Christine Chandra, M.M. Mirna, Y.
Sudaryanto, S. Ismadji: Adsorption of basic dye
onto activated carbon prepared from durian
shell: Studies of adsorption equilibrium and
kinetics. Chemical Engineering Journal, 2007,
127: 121-129.
3．Alfin Kurniawan, Vincentius Ochie Arief
Sisnandy, Kiki Trilestari, Jaka Sunarso, Nani
Indraswati, Suryadi Ismadji: Performance of
durian shell waste as high capacity biosorbent
for Cr(VI) removal from synthetic wastewater.
Ecological Engineering, 2011, 37: 940-947.
4．Xie G, Bao L, He RR, Lv YQ, Yao YX , Su YB
:Protective effects of Durian shell alcohol-
extract on liver injury in mice induced by
restraint stress. Traditional Chinese Drug
Research & Clinical Pharmacolgy, 2008, 19:
21-25.
5. Cui J, Li ZL, Hong XY: Bio-antioxidants with
ill therapy. Journal of Tsinghua university
(science and techology) , 2000, 40(6): 9-12.
6．Li XC, Wu XT, Huang L: Correlation between
Antioxidant Activities and Phenolic Contents
of Radix Angelicae Sinensis (Danggui).
Molecules, 2009, 14: 5349-5361.
7. Oyaizu M: Studies on product of browning
reaction prepared from glucose amine. Japan
Journal Nutrition, 1986, 44: 307-315.
8. Gülçin Ý: Antioxidant activity of resveratrol: A
structure-activity insight. Innovative Food

Science and Emerging Technologies, 2010, 11:
210-218.
9. Marklund S, Marklund G: Involvement of the
superoxide anion radical in the autoxidation of
pyrogallol and convenient assay for superoxide
dismutase. Eur. J. Biochem, 1974, 47: 469-474.
10. Wang XZ, Li XC, Chen DF: Evaluation of
Antioxidant Activity of Isoferulic Acid in vitro.
Natural Product Communication, 2011, 6(9):
1285-1288.
11. Wang XZ, Li XC, Li HR: Reassessment of
Antioxidant Activity of Baicalein in vitro. Asian
Journal of Pharmaceutical and Biological
Research, 2011, 1(2): 186-194.
12. Singleton VL, Rosso JA. Colorimetry of total
phenols with phosphor-molybdic-
phosphtougsitc acid reagent. Am J Enol Vit,
1965, 16: 144–158.
13. Zhu QY, Hackman RM, Ensunsa JL, Holt RR,
551
View publication stats
Asian J Pharm Biol Res Oct-Dec 2011 Vol. 1(4)
Keen CL: Antioxidative Activities of Oolong
Tea. J. Agric. Food Chem, 2002, 50: 6929-
6934.
14. Yildirim A, Mavi A, Kara AA: Determination
of antioxidant and antimicrobial activities of
Rumex crispus L. extract. Journal of
Agricultural and Food Chemistry, 2001,
49:4083-4089.
15. Jung MJ, Heo SI, Wang MH: Free radical
scavenging and total phenol contents from
methanolic extract of Ulmus davidiana, Food
Chemistry, 2008, 108: 482-487.
16. Valko M, Leibfritz D, Moncol J, Cronin MTD,
Mazur M, Telser J: Free radicals and
antioxidants in normal physiological functions
and human disease. The International Journal
of Biochemistry & Cell Biology, 2007, 39: 44-
84.