Turkish Journal of Agriculture and Forestry

Volume 32 Number 1 Article 8

1-1-2008

Antioxidant Activity of Cauliflower (Brassica oleracea L.)

EKREM KÖKSAL

İLHAMİ GÜLÇİN

Follow this and additional works at: https://journals.tubitak.gov.tr/agriculture

Part of the Agriculture Commons, and the Forest Sciences Commons

Recommended Citation
KÖKSAL, EKREM and GÜLÇİN, İLHAMİ (2008) "Antioxidant Activity of Cauliflower (Brassica oleracea L.),"
Turkish Journal of Agriculture and Forestry: Vol. 32: No. 1, Article 8. Available at:
https://journals.tubitak.gov.tr/agriculture/vol32/iss1/8

This Article is brought to you for free and open access by TÜBİTAK Academic Journals. It has been accepted for
inclusion in Turkish Journal of Agriculture and Forestry by an authorized editor of TÜBİTAK Academic Journals. For
more information, please contact [email protected].
Turk J Agric For
32 (2008) 65-78
© TÜB‹TAK

Antioxidant Activity of Cauliflower (Brassica oleracea L.)

Ekrem KÖKSAL, ‹lhami GÜLÇ‹N*

Atatürk University, Faculty of Science and Arts, Department of Chemistry, 25240 Erzurum - TURKEY

Received: 23.07.2007

Abstract: Recently, a number of studies on the health benefits associated with fruits, vegetables, herbs and spices demonstrated
that they possess potent antioxidant, anti-inflammatory, anti-mutagenic, and anti-carcinogenic activity. The potential antioxidant
activity of water and ethanol extracts of cauliflower (Brassica oleracea L.) were investigated to evaluate their potential value as a
natural ingredient for foods or cosmetic application. In this study antioxidant activity was measured by 2,2'-azino-bis(3-
ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging, 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH•) scavenging, N,N-
dimethyl-p-phenylenediamine dihydrochloride (DMPD) radical scavenging, superoxide anion (O2•–) radical scavenging, total
antioxidant activity, reducing activity using Fe+3-Fe+2 transformation and CUPRAC assays, hydrogen peroxide (H2O2) scavenging, and
ferrous metal chelating activity assays. The water extract of cauliflower (WEC) and ethanol extract of cauliflower (EEC), as
antioxidants, neutralized the activity of radicals and inhibited the peroxidation reactions of linoleic acid emulsion. Total antioxidant
activity was measured according to the ferric thiocyanate method. α-Tocopherol and trolox, a water-soluble analogue of tocopherol,
were used as the reference antioxidant compounds. WEC and EEC showed 88.6% and 80.1% inhibition of lipid peroxidation of
linoleic acid emulsion, respectively, at the concentration of 30 µg ml–1. On the other hand, at the same concentration, the standard
antioxidants α-tocopherol and trolox exhibited 68.1.4% and 81.3% inhibition of peroxidation of linoleic acid emulsion, respectively.
In addition, WEC and EEC had effective DPPH•, ABTS•+, DMPD•+, and superoxide anion radical scavenging, hydrogen peroxide
scavenging, total reducing power, and metal chelating of ferrous ion activity. Also, those various antioxidant activities were compared
to α-tocopherol and trolox as references antioxidants.

Key Words: Cauliflower, Brassica oleracea; antioxidant activity, radical scavenging

Karnabahar›n (Brassica oleracea L.) Antioxidan Aktivitesi

Özet: Son zamanlarda meyvelerin, sebzelerin, bitkilerin, otlar›n ve baharatlar›n sahip oldu¤u kuvvetli antioksidan, antienflamatuar,
antimutajenik ve antikarsinojenik aktivitelerinin sa¤l›k ile ilgi faydalar› üzerinde birçok çal›flma yap›lm›flt›r. Karnabahar›n (Brassica
oleracea L.) su ve etanol ekstrelerinin antioksidan aktiviteleri g›da ve kozmetik alandaki uygulamalar› için araflt›r›ld›. Bu çal›flmada
karnabahar›n (Brassica oleracea L.) antioksidant aktivitesi 2,2'-azino-bis(3-etilbenztiyoazolin-6-sülfonik asit) (ABTS) radikal giderme,
1,1-difenil-2-pikril-hidrazil serbest radikal (DPPH•) giderme, N,N-dimetil-p-fenilendiamin dihidroklorit (DMPD) radikal giderme,
•– +3 +2
süperoksit anyon (O2 ) radikal giderme, total antioksidan aktivite, Fe -Fe dönüflüm ve Cuprac metotlar›na göre indirgeme
aktivitesi, hidrojen peroksit (H2O2) giderme ve ferröz iyonlar› (Fe+2) kelatlama aktivitesi metotlar› ile ölçüldü. Birer antioksidan
kayna¤› olarak karnabahar›n su (WEC) ve etanol (EEC) ekstraktlar› radikalleri nötralize ve linoleik asit emülsiyonunun
peroksidasyonunu ise inhibe etti¤i gözlendi. Total antioksidan aktivite ferrik tiyosiyanat metoduna göre yap›ld›. α-Tokoferol ve
α-tokoferolün suda çözünen bir analo¤u olan troloks referans antioksidan maddeler olarak kullan›ld›. WEC ve EEC 30 µg ml–1
konsantrasyonunda, linoleik asit emülsiyonunun peroksidasyonunu s›ras›yla %88.6 ve %80.1 inhibe etti¤i belirlendi. Bunun yan›s›ra
• •+ •+ •–
WEC ve EEC etkili bir flekilde DPPH , ABTS , DMPD ve süperoksit anyon (O2 ) radikal giderme, hidrojen peroksit giderme,
+2
indirgeme kuvveti ve ferröz iyonlar›n› (Fe ) kelatlama aktivitesine de sahip oldu¤u belirlendi. Ayr›ca bu farkl› antioksidan aktiviteler
birer referans antioksidan olarak α-tokoferol ve troloks ile karfl›laflt›r›ld›.

Anahtar Sözcükler: Karnabahar, Brassica oleracea, antioksidan aktivite, radikal giderme

* Correspondence to: [email protected]

65
Antioxidant Activity of Cauliflower (Brassica oleracea L.)

Introduction recent years, the restricted use of synthetic antioxidants,

Oxygen, an element indispensable for life, can, under such as BHA and BHT, has caused increased interest in
certain circumstances, adversely affect the human body. natural antioxidant substances (Baardseth, 1989; Gülçin
Oxidation processes are very important to living et al., 2005a). Thus, development of safer natural
organisms. Most of the potentially harmful effects of antioxidants from extracts of spices and other plant
oxygen are due to the formation of reactive oxygen materials that can replace synthetic antioxidants is of
species (ROS). The uncontrolled production of ROS and interest (Liyana-Pathirana and Shahidi, 2006). Plant
the unbalanced mechanism of antioxidant protection tissues synthesize a wide variety of phenolic compounds
(Loliger, 1991). Many kinds of antioxidative components
result in the onset of many diseases and accelerate
that contain polyphenolic compounds, chlorophylls,
ageing. ROS are a class of highly reactive molecules
carotenoids, tocopherol derivatives, lignan, and related
formed during aerobic life in living organisms and include
- isoprenoids have been isolated from different kinds of
superoxide anion radicals (O2• ), hydroxyl radicals (OH•),
plants, such as oilseeds, cereal crop, vegetables, leaves,
and non free-radical species, such as H2O2 and singlet
roots, spices, herbs, and seaweeds, for use as
oxygen (1O2) (Halliwell and Gutteridge, 1989; Gülçin et
antioxidants (Wettasinghe and Shahidi, 1999, Gülçin et
al., 2002a; 2002b). There is a balance between the
al., 2006b).
generation of ROS and inactivation of ROS by the
antioxidant system in organisms. When there is imbalance Nowadays, natural antioxidants have become a major
between ROS and antioxidant defense mechanisms, ROS area of scientific research (Demo et al., 1998; Sanchez-
lead to oxidative modification in cellular membranes or Moreno et al., 1999); therefore, the importance of
intracellular molecules (Duh et al., 1999; Büyükokuroglu searching for and exploiting natural antioxidants,
et al., 2001; Gülçin et al., 2003). In addition, under especially those of plant origin, has increased greatly in
pathological conditions or oxidative stress, ROS are recent years. There is a growing interest in natural
overproduced and result in peroxidation of membrane additives as potential antioxidants (Grice, 1986; Moure et
lipids, leading to the accumulation of lipid peroxides; al., 2001; Gülçin et al., 2002a; 2006a; Oktay et al.,
however, they are removed by antioxidant defense 2003). Natural antioxidants are known to exhibit a wide
mechanisms. Antioxidants are considered as possible range of biological effects, including antibacterial,
protective agents, reducing oxidative damage from ROS antiviral, anti-inflammatory, anti-allergic, anti-
in the human body and retarding the progress of many thrombotic, and vasodilatory activity. In fact, a
chronic diseases, as well as lipid peroxidation (Pryor, fundamental property important for life is antioxidant
1991; Kinsella et al., 1993; Lai et al., 2001; Gülçin et al., activity and this property may give rise to anti-
2003a). Therefore, there is a growing interest in carcinogenicity, anti-mutagenicity, and anti-aging activity,
substances that exhibit antioxidant properties, which are among others (Cook and Samman, 1996; Liyana-
supplied to humans and animals as food components or Pathirana and Shahidi, 2006).
as specific pharmaceuticals. Antioxidants may be defined The main objectives of the present study were to
as compounds that inhibit or delay the oxidation of other assess the antioxidant potential of water and ethanol
molecules by inhibiting the initiation or propagation of extracts of cauliflower (Brassica oleracea L.) (WEC and
oxidizing chain reactions (Velioglu et al., 1998). Butylated EEC, respectively) with in vitro antioxidant assays,
hydroxyanisole (BHA), butylated hydroxytoluene (BHT), including DPPH• scavenging, ABTS•+ scavenging, DMPD•+
propyl gallate, and tert-butylhydroquinone are the most scavenging, superoxide anion radical scavenging, total
commonly used antioxidants at the present time; antioxidant activity by ferric thiocyanate method,
however, their safety has recently been questioned due to reducing power by Fe+3-Fe+2 transformation and CUPRAC
their toxicity and possible carcinogenicity (Wichi, 1988; assays, hydrogen peroxide scavenging, and metal
Sherwin, 1990; Sun and Fukuhara, 1997). Hence, in chelating effect on ferrous ions (Fe+2).

66
 E. KÖKSAL, ‹. GÜLÇ‹N

Materials and Methods described (Gülçin et al., 2003a; 2004e). Total phenolic
Chemicals content in WEC and EEC was determined as micrograms
of gallic acid equivalent using an equation obtained from
N,N-Dimethyl-p-phenylenediamine dihydrochloride the standard gallic acid graph (R2: 0.9217):
(DMPD), 2,9-dimethyl-1,10-phenanthroline (neocuproine),
2,2-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) Absorbance (λ760) = 0.014 × [Phenols (µg)]
(ABTS), riboflavin, methionine, butylated hydroxyanisole Total antioxidant activity-ferric thiocyanate
(BHA), butylated hydroxytoluene (BHT), nitroblue method
tetrazolium (NBT), the stable free radical 1,1-diphenyl-2- The antioxidant activity of WEC, EEC, and standards
picryl-hydrazyl (DPPH•), linoleic acid, 3-(2-Pyridyl)-5,6- was determined according to the ferric thiocyanate
bis (4-phenyl-sulfonic acid)-1,2,4-triazine (Ferrozine®), method in linoleic acid emulsion (Mitsuda et al., 1996), as
6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid described in a previous study (Gülçin et al., 2005b). With
(Trolox), ethylenediaminetetraacetic acid (EDTA), α- this method peroxide formation occurred during the
tocopherol, polyoxyethylene sorbitan monolaurate oxidation of linoleic acid oxidation. These compounds
(Tween-20), CuCl2, and trichloroacetic acid (TCA) were oxidized Fe2+ to Fe3+. The latter ions form a complex with
obtained from Sigma (Sigma-Aldrich GmbH, Sternheim, thiocyanate and this complex has a maximum absorbance
Germany). Ammonium thiocyanate was purchased from at 500 nm. The percent inhibition of lipid peroxidation in
Merck. All other chemicals used were analytical grade and linoleic acid emulsion was calculated by the following
obtained from either Sigma-Aldrich or Merck. equation:
Plant materials and extraction procedure
Cauliflower (Brassica oleracea L.) buds were obtained
Inhibition of lipid peroxidation (%) = 100 – A S × 100
from a local market in Erzurum, Turkey. Whole AC
cauliflower (Brassica oleracea L.) buds were cut into small
pieces and left to dry on a bench under ambient where AC is the absorbance of the control reaction and AS
temperature for over 1 week, after which they were is the absorbance in the presence of the sample of WEC,
ground in a blender. For WEC, 100 g of dried cauliflower EEC, or other test compounds. In the control, the sample
(Brassica oleracea L.) buds was ground into a fine powder was replaced with an equal volume of ethanol (Gülçin et
in a mill and mixed with 400 ml of boiling water with a al., 2004a; Gülçin, 2006b).
magnetic stirrer for 15 min. Then, the extract was 3+ 2+
filtered over Whatman No. 1 paper. The filtrates were Total reduction activity by Fe -Fe
frozen and lyophilized in a lyophilizator at a pressure of transformation
5 mm Hg at –50 °C (Labconco FreeZone 1L). The samples prepared for the ferric thiocyanate
For EEC, 100 g of dried cauliflower (Brassica oleracea method were used for this and the other antioxidant
L.) buds was ground into a fine powder in a mill and assays. The reducing activity of WEC and EEC was
mixed with ethanol. The residue was re-extracted under determined by the method of Oyaizu (1986), as
some conditions until extraction solvents became colorless previously described (Gülçin et al., 2004b). The capacity
(final volume 600 ml). The obtained extracts were of WEC and EEC to reduce the ferric-ferricyanide
filtered over Whatman No. 1 paper and the filtrate was complex to the ferrous-ferricyanide complex of Prussian
collected. Then, ethanol was removed using a rotary blue was determined by recording the absorbance at 700
evaporator (RE 100 Bibby, Stone Staffordshire England) nm after incubation. Increased absorbance of the reaction
at 50 °C to obtain dry extract. Both extracts were placed mixture indicates greater reduction capability (Elmastas
in a dark plastic bottle and stored at –20 °C until used. et al., 2006b; Gülçin et al., 2007a).
2+
Determination of total phenolic compounds Cupric ion (Cu ) reducing assay-CUPRAC assay

Total phenolic content of WEC and EEC was For determination of the reducing ability of WEC and
determined with Folin-Ciocalteu reagent, according to the EEC, the cupric ions (Cu2+) reducing power method was
method of Slinkard and Singleton (1977), as previously also used (Apak et al., 2006), with slight modification. To
test tubes containing different concentrations of WEC and

67
Antioxidant Activity of Cauliflower (Brassica oleracea L.)

EEC (10-30 µg ml-1), 0.25 ml of CuCl2 solution (0.01 M), where AC is the absorbance of the control and AS is the
0.25 ml of ethanolic neocuproine solution (7.5 × 10-3 M), absorbance in the presence of the sample of WEC, EEC,
and 0.25 ml of NH4Ac buffer solution (1 M) were added, or the standards (Elmastas et al., 2005; Gülçin, 2006a).
consecutively. The total volume was then adjusted with ABTS radical cation decolorization assay
distillated water to 2 ml and mixed well. Absorbance
The spectrophotometric analysis of ABTS•+ radical
against a reagent blank was measured at 450 nm after
scavenging activity was performed according to Re et al.
30 min.
(1999), as described in our previous study (Gülçin et al.,
2+
Metal chelating activity on ferrous ions (Fe ) 2006b). The ABTS•+ concentration (mM) in the reaction
Ferrous ion (Fe2+) chelation by WEC, EEC, and the medium was calculated from the following calibration
standards, including EDTA, was estimated by the curve determined by linear regression (R2: 0.9922):
Ferrozine® assay (Dinis et al., 1994), as explained by Absorbance (λ734 nm) = 2.744 × [ABTS ]
•+

Gülçin et al. (2005c). The results were expressed as a •+

The capability to scavenge the ABTS radical was
percentage of inhibition of the ferrozine-Fe2+ complex
calculated using the following equation:
formation. The percentage of inhibition of ferrozine-Fe2+
complex formation was calculated using the formula
given below: ABTS •+ scavenging effect (%) = A C–AS × 100
AC

Ferrous ion chelating effect (%) = A C–AS × 100 •+

AC where AC is the initial concentration of ABTS and AS is
the absorbance of the remaining concentration of ABTS•+
in the presence of scavengers (Gülçin et al., 2006b;
2+
where AC is the absorbance of the ferrozine-Fe complex Gülçin, 2006c).
and AS is the absorbance in the presence of the sample of DPPH free radical scavenging activity
WEC, EEC, or the standards (Gülçin et al., 2004b;
The DPPH free radical scavenging activity of WEC and
Elmastas et al., 2006).
EEC was measured by bleaching the purple-colored
Hydrogen peroxide scavenging activity ethanol solution of the stable DPPH radical, according to
The hydrogen peroxide scavenging ability of WEC and the Blois method (1958). This method was described
EEC was determined according to the method of Ruch et previously in our studies (Gülçin et al., 2004c; Gülçin
al. (1989). A solution of H2O2 (40 mM) was prepared in 2006b). The DPPH· concentration (mM) in the reaction
phosphate buffer (pH 7.4). WEC and EEC, at the 30 µg medium was calculated from the following calibration
2
ml-1 concentration in 3.4 ml of phosphate buffer, were curve determined by linear regression (R : 0.9974):
added to an H2O2 solution (0.6 ml, 40 mM). The Absorbance (λ517 nm) = 5.869 × 10–4 [DPPH•] + 0.0134
absorbance value of the reaction mixture was recorded at
The capability to scavenge the DPPH· radical was
230 nm. A blank solution contained the phosphate buffer
calculated using the following equation:
without H2O2. The hydrogen peroxide scavenging capacity
of WEC and EEC was calculated from the calibration
curve determined by linear regression (r2: 0.9831): DPPH• scavenging effect (%) = A C–AS × 100
AC
Absorbance (λ230) = 0.0497 × [H2O2]

The percentage of H2O2 scavenging of WEC, EEC, and

the standard compounds was calculated as: where AC is the initial concentration of the stable DPPH·
radical without the test compound and AS is the
absorbance of the remaining concentration of DPPH· in
H 2O2 scavenging effect (%) = A C–AS × 100 the presence of WEC and EEC (Gülçin et al., 2004c,
AC 2007a; Cristiane de Souza et al., 2004).

68
 E. KÖKSAL, ‹. GÜLÇ‹N

•+
Preparation of DMPD radical cation and Statistical Analysis
•+
measurement of DMPD scavenging All the analyses of total antioxidant activity were
The DMPD•+ radical scavenging activity of WEC and performed in duplicate. The other analyses were
EEC was determined according to the method described performed in triplicate. The data were recorded as means
by Fogliano et al. (1999). Different concentrations of ± standard deviation and analyzed using SPSS v.11.5 for
WEC and EEC samples (10-30 µg ml-1) were added to a Windows (SPSS Inc., Chicago). One-way analysis of
spectrophotometric cuvette and total volumes of these variance (ANOVA) was performed. Significant differences
samples were adjusted to 1 ml with distilled water. Then, between means were determined by LSD tests. P values
1 ml of DMPD•+ radical solution was directly transferred < 0.05 were regarded as significant and P values < 0.01
to the quartz cuvette. After 10-min incubation the very significant.
absorbance of the test samples was measured at 505 nm.
The DMPD•+ scavenging capacity for each tested sample
was calculated from the calibration curve determined by Results and Discussion
linear regression (R2: 0.9831):
The antioxidant activity of phenolic compounds is
Absorbance (λ505) = 0.0088 × [DMPD•+] mainly due to their redox properties, which allow them to
The percentage of DMPD•+ scavenging capability of act as reducing agents, hydrogen donors, and singlet
WEC and EEC was calculated using the following oxygen quenchers (Parr and Bolwell, 2000). Phenolic
equation: compounds are very important plant constituents because
of their scavenging ability, which is due to their hydroxyl
groups (Hatano et al. 1989). In addition, it was reported
DMPD•+ scavenging effect (%) = 1–A S × 100
AC that phenolic compounds are associated with antioxidant
activity and play an important role in stabilizing lipid
peroxidation (Yen et al., 1993). As can be seen in Table
where AC is the absorbance of the control, which contains
1, 78.6 and 175.7 mg of gallic acid equivalent of phenols
1 ml of control reaction (containing DMPD•+ solution,
was detected in 1 mg of WEC and EEC. Phenolic
except the WEC and EEC), and AS is the absorbance in the
compounds appear to be responsible for the antioxidant
presence of WEC and EEC. DMPD•+ decreases
activity of WEC and EEC; however, which components
significantly upon exposure to radical scavengers
(Fogliano et al., 1999). are responsible for the antioxidative activity of both
extracts remains unclear. Therefore, it is suggested that
Superoxide anion radical scavenging activity
further work should be performed on the isolation and
Superoxide radicals were generated according to the identification of the antioxidant constituents of WEC and
method of Beauchamp and Fridovich (1971), as described EEC.
by Zhishen et al. (1999), with slight modification.
Superoxide radicals were generated in riboflavin and
methionine, illuminated (20 W for 5 min), and assayed by Table 1. The yield of crude extract, total phenolic content, hydrogen
the reduction of NBT to form blue formazan. This peroxide scavenging, and superoxide anion radical scavenging
method was previously described (Gülçin, 2006c). The of WEC and EEC.
percentage of superoxide anion scavenged was calculated
WEC EEC
using the following formula:
Yield (%)
O•– A S × 100
2 scavenging effect (%) = 1– –1
Total phenolics (µg mg extract)
a
78.6 175.7
AC
Hydrogen peroxide scavenging (%) 51.2 41.4

where AC is the absorbance of the control and AS is the Superoxide anion scavenging (%) 57.5 43.9
absorbance in the presence of WEC, EEC, or the
a
standards (Gülçin et al., 2003, 2004d). Determined as gallic acid equivalent(GAE).

69
Antioxidant Activity of Cauliflower (Brassica oleracea L.)

For determining total phenolic content, calibration The total antioxidant activity of WEC, EEC, and the
curves were obtained using known quantities of standard standard compounds was determined by the ferric
gallic acid. Among the 2 extracts, EEC possessed more thiocyanate method in a linoleic acid system. WEC and
phenolic compounds. It was reported that phenolic EEC had strong antioxidant activity. The effects of
compounds are associated with antioxidant activity and various concentrations of WEC and EEC (from 10-30 µg
-1
play an important role in stabilizing lipid peroxidation ml ) on lipid peroxidation of linoleic acid emulsion are
(Yen et al., 1993, Gülçin, 2005). According to another shown in Figures 1 and 2. At the 30 µg ml-1
report, a very positive relationship between total phenols concentration, WEC and EEC exhibited 88.6% and
and antioxidant activity was found in many plant species 80.1% lipid peroxidation of linoleic acid emulsion,
(Velioglu et al., 1998). As seen in Figures 1 and 2 and respectively. On the other hand, at the same
Table 1, the results indicate that there wasn’t a positive concentration, α-tocopherol and trolox showed 68.1%
correlation between total antioxidant activity and total and 81.3% inhibition of peroxidation of linoleic acid
phenolic content of WEC and EEC; however, some emulsion, respectively. The results clearly showed that
authors did find a correlation between phenolic content WEC and EEC had more total antioxidant activity than
and antioxidant activity (Yen et al., 1993). trolox, similar to α-tocopherol at the same concentration
In the present study the antioxidant activity of WEC (30 µg ml-1).
and EEC were compared to α-tocopherol and its water- The CUPRAC method was developed as an antioxidant
soluble analogue, trolox. The antioxidant activity of WEC, capacity assay. This method is simultaneously cost-
EEC, α-tocopherol, and trolox was evaluated in a series effective, rapid, stable, selective, and suitable for a variety
of in vitro tests: DPPH· free radical scavenging, DMPD of antioxidants, regardless of chemical type or
radical scavenging, ABTS radical scavenging, superoxide hydrophilicity. Moreover, it was reported that the results
2+
anion radicals scavenging, total antioxidant activity by obtained from in vitro cupric ion (Cu ) reducing
ferric thiocyanate method, reducing activities, hydrogen measurements may be more efficiently extended to the
peroxide scavenging activity, and metal chelating activity. possible in vivo reactions of antioxidants. The CUPRAC
The ferric thiocyanate method measures the amount chromogenic redox reaction is carried out at a pH 7.0,
of peroxides produced during the initial stages of close to the physiological pH (Apak et al., 2004). This
oxidation, which are the primary products of oxidation. method is capable of measuring thiol-type antioxidants,

2.5 Control
Trolox-30 µg ml-1
α-Tocopherol-30 µg ml-1
2 WEC-10 µg ml-1
WEC-20 µg ml-1
Absorbance (500 nm)

WEC-30 µg ml-1

1.5

0.5

0
0 10 20 30 40 50 60
Time (Hour)

Figure 1. Total antioxidant activity of WEC at different concentrations (10-30 µg ml-1), and α-tocopherol and trolox
(30 µg ml-1).

70
 E. KÖKSAL, ‹. GÜLÇ‹N

2.5 Control
Trolox-30 µg ml-1
α-Tocopherol-30 µg ml-1
2
EEC-10 µg ml-1
Absorbance (500 nm)

EEC-20 µg ml-1
EEC-30 µg ml-1
1.5

0.5

0
0 10 20 30 40 50 60

Time (Hour)

Figure 2. Total antioxidant activity of EEC at different concentrations (10-30 µg ml-1), and α-tocopherol and trolox
(30 µg ml-1).

such as glutathione and non-protein thiol, unlike the Figure 3 depicts the reducing activity of WEC, EEC,
widely applied FRAP test, which is non-responsive to -SH and the standards (α-tocopherol and trolox) using the
group antioxidants. The reducing capacity of a compound potassium ferricyanide reduction method. For measuring
may serve as a significant indicator of its potential reductive activity, the Fe3+-Fe2+ transformation was
antioxidant activity (Gülçin 2006b, 2006c). In this assay, investigated in the presence of WEC and EEC, using the
the yellow color of the test solution changes to various method of Oyaizu (1986). The reducing activity of WEC,
shades of green and blue, depending on the reducing EEC, α-tocopherol, and trolox increased with increasing
power of the antioxidant sample (Chung et al., 2002; sample concentration. As can be seen in Figure 3, WEC
Gülçin and Dafltan, 2007). and EEC showed greater effective reducing activity than

1.5
Trolox
α-Tocopherol
1.2 WEC
EEC
Absorbance (700 nm)

0.9

0.6

0.3

0
0 10 20 30
Concentration (µg ml-1)

Figure 3. The Fe3+-Fe2+ reducing activity of different concentrations (10-30 µg ml-1) of WEC, EEC, α-tocopherol, and trolox.

71
Antioxidant Activity of Cauliflower (Brassica oleracea L.)

the control, at different concentrations (R2: 8608, R2: reducing capability of WEC and EEC, as determined by
7202). These differences were statistically significant (P the CUPRAC method, was concentration dependent (10-
< 0.01). The reducing power of WEC, EEC, and the 30 µg ml-1). The cupric ion (Cu2+) reducing power of
standard compounds exhibited the following order: trolox WEC, EEC and the standard compounds at the same
> α-tocopherol > WEC ≈ EEC. concentration (30 µg ml-1) exhibited the following order:
α-tocopherol > trolox > EEC ≈ WEC.
The cupric ion (Cu2+) reducing ability (CUPRAC
method) of WEC and EEC is shown in Figure 4. A The ferrous ion chelating activity of WEC, EEC, α-
correlation between the cupric ion (Cu2+) reducing ability tocopherol, trolox, and EDTA are shown in Figure 5. The
and concentrations was observed. The cupric ion (Cu2+) ferrous ion chelating activity of WEC, EEC, and the

1 Trolox
α-Tocopherol
WEC
0.75 EEC
Absorbance (450 nm)

0.5

0.25

0
0 10 20 30
Concentration (µg ml-1)

Figure 4. The Cu2+-Cu+ reducing activity of WEC, EEC, α-tocopherol, and trolox at different concentrations
(10-30 µg ml-1).

1.25

EDTA
Trolox
1 α-Tocopherol
Metal chelating (562 nm)

WEC
EEC
0.75

0.5

0.25

0
0 10 20
Concentration (µg ml-1)
Figure 5. The ferrous ion (Fe ) chelating activity of WEC, EEC, α-tocopherol, trolox, EDTA at
2+

different concentrations (10-20 µg ml-1).

72
 E. KÖKSAL, ‹. GÜLÇ‹N

standards was determined according to the method of The ability of WEC and EEC to scavenge hydrogen
Dinis et al. (1994). Among the transition metals, iron is peroxide was determined according to the method of Ruch
known to be the most important lipid oxidation pro- et al. (1989) (Table 1) and was compared to that of α-
oxidant due to its high reactivity. The ferrous state of tocopherol and trolox as standards. WEC and EEC exhibited
iron accelerates lipid oxidation by breaking down a 51.2% and 41.4% scavenging effect of hydrogen
hydrogen and lipid peroxides to reactive free radicals via peroxide at the 30 µg ml-1 concentration, respectively. On
the Fenton reaction (Fe2+ + H2O2 → Fe3+ + OH¯ + OH·). the other hand, α-tocopherol and trolox exhibited a 39.1%
Fe3+ ions also produce radicals from peroxides, although and 37.7% hydrogen peroxide scavenging activity at the
the rate is 10-fold less than that of Fe2+ ions (Miller, same concentration, respectively. These results show that
1996). Fe2+ ions are the most powerful pro-oxidant WEC and EEC have greater hydrogen peroxide scavenging
among the various species of metal ions (Halliwell and activity. At the above concentration, the hydrogen peroxide
Gutteridge, 1984; Gülçin, 2007). scavenging effect of WEC, EEC, and both standards
In fact, as shown in Figure 5, WEC and EEC disrupted decreased in the order of WEC > EEC ≈ α-tocopherol >
2+ trolox. Hydrogen peroxide itself is not very reactive;
the Fe -ferrozine complex at different concentrations
however, it can sometimes be toxic to cells because it may
(10-20 µg ml-1). The difference between all WEC and EEC
give rise to hydroxyl radical in the cells.
concentrations and the control was statistically significant
(P < 0.01). In addition, WEC and EEC exhibited 88.6% The improved technique for the generation of ABTS•+
and 89.5% chelation of ferrous ions, respectively, at the described herein involves the direct production of the
-1
20 µg ml concentration. On the other hand, the blue/green ABTS•+ chromophore through the reaction
between ABTS and potassium persulfate. As seen in
percentage of metal chelating capacity of the same
Figure 6, the ABTS•+ radical scavenging activity of WEC
concentration of α-tocopherol, trolox, and EDTA were
and EEC was concentration dependent (10-20 µg ml-1).
90.8%, 49.3% and 91.2%, respectively. The metal
There was a significant decrease (P < 0.01) in the
scavenging effect of those samples decreased in the order
concentration of ABTS•+ due to the scavenging capacity of
of EDTA ≈ α-tocopherol ≈ EEC ≈ WEC > trolox. The data WEC, EEC, and the standards. In addition, the ABTS•+
obtained from Figure 5 reveal that WEC and EEC scavenging effect of WEC, EEC, and the standards
demonstrated a marked capacity for iron binding, decreased accordingly: trolox > WEC > EEC > α-
suggesting that their main action as a peroxidation tocopherol, which was 90.7%, 79.8%, 68.3%, and
protector may be related to their iron binding capacity. 55.9% at the concentration of 20 µg ml-1, respectively.

0.7
Trolox
α-Tocopherol
WEC
0.525 EEC
Absorbance (734 nm)

0.35

0.175

0
0 10 20
Concentration (µg ml-1)

Figure 6. The stable ABTS scavenging effect of WEC, EEC, α-tocopherol, and trolox at
•+

different concentrations (10-30 µg ml-1).

73
Antioxidant Activity of Cauliflower (Brassica oleracea L.)

Antioxidants react with DPPH•, which is a stable free The principle of the DMPD assay is that at an acidic pH
radical, and convert it to 1,1-diphenyl-2-picryl hydrazine. and in the presence of a suitable oxidant solution DMPD
The degree of discoloration indicates the radical- can form a stable and colored radical cation (DMPD•+).
•+
scavenging potential of the antioxidant (Singh et al., DMPD has a maximum absorbance at 505 nm.
2002). The antioxidant activity of WEC, EEC, and the Inhibition of the absorbance at 505 nm was linear
standard antioxidants, was determined using the DPPH• -1
between 10 and 30 µg ml of WEC and EEC. This assay
method. Since the DPPH• assay can accommodate a large is based on the extent of radical cation reduction at a
number of samples in a short period of time and is fixed time point and not on the rate of reduction. This
sensitive enough to detect natural compounds at low feature eliminated the complications due to the
concentrations, it was used in the present study for the monitoring of color inhibition over time, which is present
primary screening of WEC and EEC free radical- in other methods (Pryor et al., 1993; Tubaro et al.,
scavenging activity. WEC and EEC exhibited marked
1996), and allows the simultaneous analysis of many
DPPH free radical scavenging activity in a concentration-
samples.
dependent manner. Figure 7 illustrates a significant
•+
decrease (P < 0.05) in the concentration of the DPPH DMPD scavenging efficiency was determined
radical due to the scavenging ability of WEC, EEC, and the according to Miller et al. (1996). As seen in Figure 8, the
•+
standards. α-Tocopherol and trolox were used as DMPD scavenging activity of WEC and EEC was
-1
reference radical scavengers. The DPPH radical concentration dependent (10-30 µg ml ). WEC and EEC
•+
scavenging effect of WEC, EEC, and the standards demonstrated marked DMPD scavenging activity. The
•+
decreased in the following order: EEC > trolox > α- DMPD scavenging activity of WEC, EEC, and the
tocopherol ≈ WEC, which was 64.6%, 56.5%, 51.9%, standards decreased in the following order: Trolox > EEC
and 51.2%, at the concentration of 30 µg ml-1, > WEC (α-tocopherol, a hydrophobic standard
respectively. antioxidant, was not used as a standard in this method).

2.1
Trolox
α-Tocopherol
WEC
1.75
EEC
Absorbance (517 nm)

1.4

1.05

0.7
0 10 20 30
Concentration (µg ml-1)

Figure 7. The DPPH• scavenging effect of WEC, EEC, α-tocopherol, and trolox at different
concentrations (10-30 µg ml-1).

74
 E. KÖKSAL, ‹. GÜLÇ‹N

0.8
Trolox
WEC
EEC
0.6
Absorbance (505 nm)

0.4

0.2

0
0 10 20 30
Concentration (µg ml-1)

Figure 8. The DMPD•+ scavenging effect of WEC, EEC, and trolox at different concentrations (10-30 µg ml-1).

The superoxide anion radical included in free radical Conclusion

species is a factor that can induce aging and destruct cell
This study demonstrated the potential antioxidant
membranes, and it can be generated by oxidative stress.
properties of WEC and EEC. According to the presented
In this method, superoxide anion is derived from
data, WEC and EEC were effective antioxidants in
dissolved oxygen by riboflavin-methionine- illuminate
reaction and reduces the yellow dye (NBT2+) to produce different in vitro assays, including the ferric thiocyanate
3+ 2+ 2+ +
the blue formazan, which is measured method, Fe -Fe and Cu -Cu reducing power assays,
•+
spectrophotometrically at 560 nm. Antioxidants are able DPPH• scavenging, ABTS scavenging, DMPD•+
to inhibit blue NBT formation (Cos et al., 1998; Parejo et scavenging, superoxide anion radical scavenging,
al., 2002). The decrease of absorbance at 560 nm with hydrogen peroxide scavenging, and metal chelating
antioxidants indicates the consumption of superoxide activity, when compared to the standard antioxidant
anion in the reaction mixture. Table 1 shows the percent compounds α-tocopherol (a natural antioxidant) and
inhibition of superoxide radical generation by WEC, EEC trolox (the water-soluble analogue of tocopherol). On the
and the standards at the 30 µg ml-1 concentration. As basis of the results of this study, it is clear that both plant
seen in Table 1, the percent inhibition of superoxide anion extracts have powerful antioxidant activity against various
radical generation by the 30 µg ml-1 concentration of
antioxidant systems in vitro. Moreover, WEC and EEC can
WEC and EEC was 57.5% and 43.9%, respectively. On
be used as easily accessible sources of natural antioxidants
the other hand, at the same concentration, α-tocopherol
and possible food supplements, or in pharmaceutical
and trolox exhibited 21.3% and 23.9% superoxide anion
radical scavenging activity, respectively. applications.

References
Apak, R., K. Güçlü, M. Özyürek and S.E. Karademir. 2004. A novel total Apak, R., K. Güçlü, M. Özyürek, S.E. Karademir and E. Erça. 2006. The
antioxidant capacity index for dietary polyphenols, vitamin C and cupric ion reducing antioxidant capacity and polyphenolic content
E, using their cupric ion reducing capability in the presence of of some herbal teas. Int. J. Food Sci. Nut. 57: 292-304.
neocuproine: The CUPRAC method. J. Agric. Food Chem. 52:
7970-7981.

75
Antioxidant Activity of Cauliflower (Brassica oleracea L.)

Arabshahi-Delouee S. and A. Urooj. 2007. Antioxidant properties of Finefrock, A.E., A.I. Bush and P.M. Doraiswamy. 2003. Current status
various solvent extracts of mulberry (Morus indica L.) leaves. of metals as therapeutic targets in Alzheimer’s disease. J. Am.
Food Chem. 102: 1233-1240. Geriat. Soc. 51: 1143-1148.
Baardseth, P. 1989. Effect of selected antioxidants on the stability of Fogliano, V., V. Verde, G. Randazzo and A. Ritieni. 1999. Method for
dehydrated mashed potatoes. Food Addit. Contam. 6: 201-207. measuring antioxidant activity and its application to monitoring
the antioxidant capacity of wines. J. Agric. Food Chem. 47: 1035-
Bast, A., G.R.M.M. Haenen and C.J.A. Doelman. 1991. Oxidants and
1040.
antioxidants: state of the art. Am. J. Med. 91: 2-13.
Grice, H.C. 1986. Safety evaluation of butylated hydroxytoluene (BHT)
Beauchamp, C. and I. Fridovich. 1971. Superoxide dismutase: improved
in the liver, lung and gastrointestinal tract. Food Chem. Toxicol.
assays and an assay applicable to acrylamide gels. Anal. Biochem.
24: 1127-1130.
44: 276-287.
Gülçin, ‹. 2005. The antioxidant and radical scavenging activities of
Blois, M.S. 1958. Antioxidant determinations by the use of a stable free
black pepper (Piper nigrum) seeds. Int. J. Food Sci. Nut. 56: 491-
radical. Nature 26, 1199-1200
499.
Brand-Williams, W., M.E. Cuvelier and C. Berset. 1995. Use of a free
Gülçin, ‹. and A. Dafltan. 2007. Synthesis of dimeric phenol derivatives
radical method to evaluate antioxidant activity. Lebensm. Wiss.
and determination of in vitro antioxidant and radical scavenging
Techonol. 28: 25-30.
activities. J. Enzym. Inhib. Med. Chem. 22: 685-695.
Büyükokuro¤lu, M.E., ‹. Gülçin, M. Oktay and Ö.‹. Küfrevio¤lu. 2001.
Gülçin, ‹. 2006b. Antioxidant activity of caffeic acid (3,4-
In vitro antioxidant properties of dantrolene sodium. Pharmacol.
dihydroxycinnamic acid). Toxicology 217: 213-220.
Res. 44: 491-495.
Gülçin, ‹. 2006c. Antioxidant and antiradical activities of L-carnitine. Life
Chung, Y.C., C.T. Chang, W.W. Chao, C.F. Lin and S.T. Chou. 2002.
Sci. 78: 803-811.
Antioxidative activity and safety of the 50% ethanolic extract
from red bean fermented by Bacillus subtilis IMR-NK1. J. Agric. Gülçin, ‹., D. Berashvili and A. Gepdiremen. 2005a. Antiradical and
Food Chem. 50: 2454-2458. antioxidant activity of total anthocyanins from Perilla pankinensis
decne. J. Ethnopharmacol. 101: 287-293.
Cook, N.C. and S. Samman. 1996. Flavonoids: chemistry, metabolism,
cardioprotective effects and dietary sources. J. Nut. Biochem. 7: Gülçin, ‹., fi. Beydemir, H.A. Alici, M. Elmastafl and M.E. Büyükokuro¤lu.
66-76. 2004c. In vitro antioxidant properties of morphine. Pharmacol.
Res. 49: 59-66.
Cos, P., L.Y. Ying, M. Calomme, J.H. Hu, K. Cimanga, B. Van Poel, L.
Pieters, A.J. Vlietinck and D.V. Berghe. 1998. Structure activity Gülçin, ‹., M.E. Büyükokuro¤lu, M. Oktay and Ö.‹. Küfrevio¤lu. 2003a.
relationships and classification of flavonoids as inhibitors of Antioxidant and analgesic activities of turpentine of Pinus nigra
xanthine oxidase and superoxide scavengers. J. Nat. Prod. 61: Arn. Subsp. pallsiana (Lamb.) Holmboe. J. Ethnopharmacol. 86:
71-76. 51-58.
Demo, A., P. Kefalas and D. Boskou. 1998. Nutrient antioxidants in Gülçin, ‹., M.E. Büyükokuro¤lu, M. Oktay and Ö.‹. Küfrevio¤lu. 2002a.
some herbs and Mediterranean plant leaves. Food Res. Int. 32: On the in vitro antioxidant properties of melatonin. J. Pineal Res.
351–354 33: 167-171.
Dinis, T.C.P., V.M.C. Madeira and L.M. Almeida. 1994. Action of Gülçin, ‹. 2007. Comparison of in vitro antioxidant and antiradical
phenolic derivates (acetoaminophen, salycilate, and 5- activities of L-tyrosine and L-Dopa. Amino Acids. 32: 431-438.
aminosalycilate) as inhibitors of membrane lipid peroxidation and
Gülçin, ‹., R. Elias, A. Gepdiremen and L. Boyer. 2006b. Antioxidant
as peroxyl radical scavengers. Arch. Biochem. Biophys. 315: 161-
activity of lignans from fringe tree (Chionanthus virginicus L.).
169.
Eur. Food Res. Technol. 223: 759-767.
Duh, P.D., Y.Y. Tu and G.C. Yen. 1999. Antioxidant activity of water
Gülçin, ‹., M. Elmastas and H.Y. Aboul-Enein. 2007a. Determination of
extract of harng jyur (Chrysanthemum morifolium Ramat).
antioxidant and radical scavenging activity of basil (Ocimum
Lebensm. Wiss. Technol. 32: 269-277.
basilicum) assayed by different methodologies. Phytother. Res.
Elmastafl, M., ‹. Gülçin, Ö. Beydemir, Ö.‹. Küfrevio¤lu and H.Y. Aboul- 21: 354-361.
Enein. 2006. A study on the in vitro antioxidant activity of juniper
Gülçin, ‹., H.A. Alici, M. Cesur. 2005b. Determination of in vitro
(Juniperus communis L.) seeds extracts. Anal. Lett. 39: 47-65
antioxidant and radical scavenging activities of propofol. Chem.
Elmastas, M., ‹. Gülçin, Ö. Iflıldak, Ö.‹. Küfrevio¤lu, K. ‹bao¤lu and H.Y. Pharm. Bull. 53: 281-285.
Aboul-Enein. 2006b. Antioxidant capacity of bay (Laurus nobilis
Gülçin, ‹., fi. Beydemir, ‹.G. fiat, Ö.‹. Küfrevio¤lu. 2005c. Evaluation of
L.) leave e extracts. J. Iran. Chem. Soc. 3: 258-266.
antioxidant activity of cornelian cherry (Cornus mas L.). Acta
Elmastafl, M., ‹. Gülçin, L. Öztürk and ‹. Gökçe. 2005. Investigation of Aliment. Hung. 34: 193-202.
antioxidant properties of spearmint (Mentha spicata L.). Asian J.
Chem. 17: 137-148.

76
 E. KÖKSAL, ‹. GÜLÇ‹N

Gülçin, ‹., Ö.‹. Küfrevio¤lu, M. Oktay and M.E. Büyükokuro¤lu. 2004d. Miller, J.N., C.A. Rice-Evans and M.J. Davies. 1993. A novel method for
Antioxidant, antimicrobial, antiulcer and analgesic activities of measuring antioxidant capacity and its application to monitoring
nettle (Urtica dioica L.). J. Ethnopharmacol. 90: 205-215. the antioxidant status in premature neonates. Clin. Sci. 84: 407-
412.
Gülçin, ‹., V. Mshvildadze, A. Gepdiremen and R. Elias. 2006a.
Screening of antioxidant and antiradical activity of Miller, N.J., J. Sampson, L.P. Candeias, P.M. Bramley and C.A. Rice-
monodesmosides and crude extract from Leontice smirnowii Evans. 1996. Antioxidant activities of carotenes and xanthophylls.
Tuber. Phytomedicine 20: 130-134 FEBS Lett. 384: 240-242.
Gülçin, ‹., M. Oktay, E. Kireçci and Ö.‹. Küfrevio¤lu. 2003. Screening of Min, D.B. 1998. Lipid oxidation of edible oil. In: Food Lipids chemistry,
antioxidant and antimicrobial activities of anise (Pimpinella anisum nutrition, and biotechnology. (Eds: C.C. Akoh, Min, D.B.), Marcel
L.) seed extracts. Food Chem. 83: 371-382. Dekker, New York, pp. 283-296.
Gülçin, ‹., M. Oktay, Ö.‹. Küfrevio¤lu and A. Aslan. 2002b. Mitsuda, H., K. Yuasumoto and K. Iwami. 1996. Antioxidation action of
Determination of antioxidant activity of lichen Cetraria islandica indole compounds during the autoxidation of linoleic acid. Eiyo to
(L) Ach. J. Ethnopharmacol. 79: 325-329. Shokuryo, 19: 210-214.
Gülçin, ‹., ‹.G. fiat, fi. Beydemir and Ö.‹. Küfrevio¤lu. 2004b. Evaluation Moure, A., J.M. Cruz, D. Franco, J.M. Dominguez, J. Sineiro, H.
of the in vitro antioxidant properties of extracts of broccoli Dominguez, M.J. Nunez and J.C. Parajo. 2001. Natural
(Brassica oleracea L.). Ital. J. Food Sci. 16: 17-30. antioxidants from residual sources. Food Chem. 72: 145-171.
Gülçin, ‹., ‹.G. fiat, fi. Beydemir, M. Elmastafl and Ö.‹. Küfrevio¤lu. Oktay, M.,, ‹. Gülçin and Ö.‹. Küfrevio¤lu. 2003. Determination of in
2004a. Comparison of antioxidant activity of clove (Eugenia vitro antioxidant activity of fennel (Foeniculum vulgare) seed
caryophylata Thunb) buds and lavender (Lavandula stoechas L.). extracts. Lebensm. Wiss. Technol. 36: 263-271.
Food Chem. 87: 393-400.
Oyaizu, M. 1986. Studies on product of browning reaction prepared
Gülçin, ‹., M.T. U¤uz, M. Oktay, fi. Beydemir and Ö.‹. Küfrevio¤lu. from glucose amine. Jpn. J. Nut. 44: 307-315.
2004e. Evaluation of antioxidant and antimicrobial activities of
Parejo, I., F. Viladomat, J. Bastida, A. Rosas-Romero, N. Flerlage, J.
clary sage (Salvia sclarea L.). Turk. J. Agric. For. 28: 25-33.
Burillo and C. Codina. 2002. Comparison between the radical
Haber, F. and J. Weiss. 1934. The catalytic decomposition of hydrogen scavenging activity and antioxidant activity of six distilled and
peroxide by iron salts. Proc. Roy. Soc. London Ser A, 147: 332- nondistilled Mediterranean herbs and aromatic plants. J. Agric.
351. Food Chem. 50: 6882-6890.
Halliwell, B. and J.M.C. Gutteridge. 1989. Free radicals in biology and Park, Y.K., M.H. Koo, M. Ikegaki and J.L. Contado. 1997. Comparison
medicine. Clarendon Press, Oxford, pp. 23-30. of the flavonoid aglycone contents of Apis mellifera propolis from
various regions of Brazil. Arquivos de Biologiae Technologia 40:
Halliwell, B. and J.M.C. Gutteridge. 1984. Oxygen toxicology, oxygen
97-106.
radicals, transition metals and disease. Biochem. J. 219: 1-4.
Parr A. and G.P. Bolwell. 2000. Phenols in the plant and in man: The
Halliwell, B. 1991. Reactive oxygen species in living systems: source,
potential for possible nutritional enhancement of the diet by
biochemistry and role in human disease. Am. J. Med. 91: 14-22.
modifying the phenols content or profile. J. Sci. Food Agric. 80:
Hatano, T., R. Edamatsu, A. Mori, Y. Fujita and E. Yasuhara. 1989. 985-1012.
Effect of interaction of tannins with co-existing substances. VI.
Pietta, P.G. 2000. Flavonoids as antioxidants. J. Nat. Prod. 63: 1035-
Effects of tannins and related polyphenols on superoxide anion
1042.
radical and on DPPH radical. Chem. Pharm. Bull. 37: 2016-2021.
Pryor, W.A., J.A. Cornıcelli, L.J. Devall, B. Tait, B.K. Trivedi, D.T. Witiak
Kehrer, J.P. 2000. The Haber-Weiss reaction and mechanisms of
and M. Wut. 1993. A rapid screening test to determine the
toxicity. Toxicology 149: 43-50.
antioxidant potencies of natural and synthetic antioxidants. J.
Kinsella, J.E., E. Frankel, B. German and J. Kanner. 1993. Possible Org. Chem. 58: 3521-3522.
mechanism for the protective role of the antioxidant in wine and
Pryor, W.A. 1991. The antioxidant nutrient and disease prevention-
plant foods. Food Technol. 47: 85-89.
What do we know and what do we need to find out? Am. J. Clin.
Lai, L.S., S.T. Chou and W.W. Chao. 2001. Studies on the antioxidative Nut. 53: 391-393.
activities of Hsian-tsao (Mesona procumbens Hemsl) leaf gum. J.
Re, R., N. Pellegrini, A. Proteggente, A. Pannala, M. Yang and C. Rice-
Agric. Food Chem. 49: 963-968.
Evans. 1999. Antioxidant activity applying an improved ABTS
Liyana-Pathirana, C.M. and F. Shahidi. 2006. Antioxidant properties of radical cation decolorization assay. Free Radical Bio. Med. 26:
commercial soft and hard winter wheats (Triticum aestivum L.) 1231-1237.
and their milling fractions. J. Sci. Food Agric. 86: 477-485.
Ruch, R.J., S.J. Cheng and J.E. Klaunig. 1989. Prevention of
Loliger, J. 1991. The use of antioxidants in foods. In: Free Radicals and cytotoxicity and inhibition of intracellular communication by
Food Additives. (Eds. O.I., Auroma, B., Halliwell,), Taylor & antioxidant catechins isolated from Chinese green tea.
Francis, London, pp. 129-150. Carcinogenesis 10: 1003-1008.

77
Antioxidant Activity of Cauliflower (Brassica oleracea L.)

Sanchez-Moreno, C., J.A. Larrauri and F. Saura-Calixto. 1999. Free Wichi, H.P. 1988. Enhanced tumor development by butylated
radical scavenging capacity and inhibition of lipid oxidation of hydroxyanisole (BHA) from the prospective of effect on
wines, grape juices and related polyphenolic constituents. Food forestomach and oesophageal squamous epithelium. Food Chem.
Res. Int. 32: 407-412. Toxicol. 26: 717-723.
Sherwin, E.R. 1990. Food additives. Branen, A.L.; Davidson, P.M. Wickens, A.P. 2001. Aging and the free radical theory. Reportor.
Salminen, S., Marvel Dekker inc, New York, pp. 139-193. Physiol. 128:379-391.
Singh, R.P., K.N.C. Murthy and G.K. Jayaprakasha. 2002. Studies on Wolfenden, B.S. and R.L. Willson. 1982. Radical-cations as reference
the antioxidant activity of pomegranate (Punica granatum) peel chromogens in kinetic studies of one-electron transfer reactions:
and seed extracts using in vitro models. J. Agric. Food Chem. 50: pulse radiolysis studies of 2,2'-azinobis-(3-ethylbenzthiazoline-6-
81-86. sulphonate). J. Chem. Soc. Perk. T. 2: 805-812.
Slinkard, K. and V.L. Singleton. 1977. Total phenol analyses: Wong, P.Y.Y. and D.D. Kitts. 2001. An iron binding assay to measure
Automation and comparison with manual methods. Am. J. Enol. activity of known food sequestering agents: studies with
Viticult. 28: 49-55. buttermilk solids. Food Chem. 72: 245-254.
Strlic, M., T. Radovic, J. Kolar and B. Pihlar. 2002. Anti- and Yamaguchi, F., T. Ariga, Y. Yoshimira and H. Nakazawa. 2000.
prooxidative properties of gallic acid in Fenton-type systems. J. Antioxidant and antiglycation of carcinol from Garcinia indica fruit
Agr. Food Chem. 50: 6313-6317. rind. J. Agric. Food Chem. 48: 180-185.
Sun, B. and M. Fukuhara. 1997. Effects of co-administration of Yen, G.C., P.D. Duh and C.L. Tsai. 1993. Relationship between
butylated hydroxytoluene, butylated hydroxyanisole and antioxidant activity and maturity of peanut hulls. J. Agric. Food
flavonoids on the activation of mutagens and drug metabolizing Chem. 41: 67-70.
enzymes in mice. Toxicology 122: 61-72.
Yen, G.C. and P.D. Duh. 1994. Scavenging effect of methanolic extract
Tubaro, F., E. Micossi and F. Ursini. 1996. The antioxidant capacity of of peanut hulls on free radical and active oxygen species. J. Agric.
complex mixtures by kinetic analysis of crocin bleaching inhibition. Food Chem. 42: 629-632.
JAOCS. 73: 173-179.
Zhishen, J., T. Mengcheng and W. Jianming. 1999. The determination
Velioglu, Y.S., G. Mazza, L. Gao and B.D. Oomah. 1998. Antioxidant of flavonoid contents on mulberry and their scavenging effects on
activity and total phenolics in selected fruits, vegetables and grain superoxide radical. Food Chem. 64: 555-559.
products. J. Agric. Food Chem. 46: 4113-4117.
Zhu, Q.Y., R.M. Hackman, J.L. Ensunsa, R.R. Holt and C.L. Keen. 2002.
Wettasinghe, M. and F. Shahidi. 1999. Antioxidant and free radical- Antioxidative activities of oolong tea. J. Agric. Food Chem. 50:
scavenging properties of ethanolic extracts of defatted borage 6929-6934.
(Borago officinalis L.) seeds. Food Chem. 67: 399-414.

78