Antioxidant and Antiradical Activities of Flavonoids

Stani sl aw Burda* and Wi esl aw Ol eszek

Department of Bi ochemi stry, I nsti tute of Soi l Sci ence and Pl ant Cul ti vati on, ul . Czartoryski ch 8,
24-100 Pul awy, Pol and
The rel ati onshi p between the structure of 42 fl avonoi ds and thei r anti oxi dant and anti radi cal
acti vi ti es was el uci dated by heat-i nduced oxi dati on i n a -carotene and l i nol ei c aci d system and by
the 1,1-di phenyl -2-pi cryl hydrazyl decol orati on test. From seven structural l y di vergent groups of
fl avonoi ds, onl y fl avonol s wi th a free hydroxyl group at the C-3 posi ti on of the fl avonoi d skel eton
showed hi gh i nhi bi tory acti vi ty to -carotene oxi dati on. Anti radi cal acti vi ty depended on the presence
of a fl avonol structure or free hydroxyl group at the C-4 posi ti on. The effect of the 4-hydroxyl was
strongl y modi fi ed by other structural features, such as the presence of free hydroxyl s at C-3 and/or
C-3 and a C2-C3 doubl e bond.
Keywords: Flavonoids; antioxidant activity; antiradical activity
I NTRODUCTI ON
Fl avonoi ds, deri vati ves of benzo--pyrone, are wi de-
spread i n pl ants. Several paper (1-8) have i ndi cated
that these compounds have the property of i nhi bi ti ng
autoxi dati on reacti ons and scavengi ng of free radi cal s,
but the rel ati onshi p between thei r structure and acti vi ty
remai ns uncl ear. Fl avonoi ds may possess mul ti pl e
properti es for scavengi ng reacti ve oxygen and ni trogen
speci es (1-3). The presence of an ortho-hydroxyl ati on
on the B-ri ng of the fl avonoi d mol ecul e, the number of
free hydroxyl groups, a C2-C3 doubl e bond i n the
C-ri ng, or the presence of a 3-hydroxyl group i s usual l y
l i sted as a condi ti on of anti oxi dant and anti radi cal
acti vi ti es (4-8).
The ai m of thi s study was to el uci date the rel ati on-
shi p between the chemi cal structure of the fl avonoi ds
and thei r abi l i ty to i nhi bi t oxi dati on i n a -carotene-
l i nol ei c aci d model system and thei r effecti veness as
the scavengers of 1,1-di phenyl -2-pi cryl hydrazyl (DPPH)
free radi cal s. The study i nvol ved seven structural l y
di fferent fl avonoi d groups: fl avones, fl avonol s, fl avan-
ones, di hydrofl avonol s, i sofl avones, bi fl avanones, and
one coumestan, a coumestrol .
MATERI ALS AND METHODS
Materials. The majori ty of the fl avonoi ds were i sol ated i n
our l aboratory (9-15). Kaempferol and nari ngeni n were
obtai ned by aci d hydrol ysi s of thei r gl ycosi des (kaempferol 3,7-
di rhamnosi de and nari ngi n, respecti vel y). The permethyl
deri vati ves (3,5,7,3,4-pentamethoxyfl avone and 3,5,7,3,4,5-
hexamethoxyfl avone) were prepared from querceti n and l ari -
cytri n by methyl ati on wi th di methyl sul fate (9). The remai ni ng
compounds were purchased from Fl uka AG (fl avone, nari ngi n,
gal angi n, and kaempferi de), Si gma Chemi cal Co. (mori n,
hespereti n, 5,7-di hydroxyfl avone, BHT, and D,L-R-tocopherol
and i ts acetate), and I CN Pharmaceuti cal s, I nc. (fl avanone,
3-hydroxyfl avone, 7-hydroxyfl avone, 8-methoxyfl avone, 5-hy-
droxyfl avone, api geni n and robi neti n).
Antioxidant Activity Determination. Heat-i nduced oxi -
dati on of an aqueous emul si on system of -carotene and
l i nol ei c aci d was used as the anti oxi dant acti vi ty test model
(16). One mi l l i l i ter of -carotene (0.2 mg/mL) di ssol ved i n
chl oroform was added to an Erl enmeyer fl ask contai ni ng
l i nol ei c aci d (0.02 mL) and Tween 20 (0.2 mL). The mi xture
was than dosed wi th 0.2 mL of the correspondi ng fl avonoi d or
standard sol uti on i n methanol at a concentrati on of 10
-3
M.
Fi fty mi l l i l i ters of di sti l l ed water, saturated for 15 mi n wi th
oxygen, was added to the fl ask. The resul ti ng mi xture was
shaken and kept for 2 h at 50 C. The absorbance of the
sampl es was measured on a Hewl ett-Packard 8453 spectrom-
eter at 470 nm, i mmedi atel y after thei r preparati on (t )0 mi n)
and at the end of the experi ment (t ) 120 mi n). Anti oxi dant
acti vi ty was cal cul ated as percent i nhi bi ti on of oxi dati on versus
control sampl e wi thout fl avonoi d added, usi ng the equati on
where A
s
0
i s the absorbance of sampl e at 0 mi n, As
120
i s the
absorbance of sampl e at 120 mi n, Ac
0
i s the absorbance of
control sampl e at 0 mi n, and Ac
120
i s the absorbance of control
sampl e at 120 mi n.
Antiradical Activity Determination. Scavengi ng free
radi cal potenti al s were tested i n a methanol i c sol uti on of
DPPH (17). The degree of decol orati on of the sol uti on i ndi cates
the scavengi ng effi ci ency of the added substance. For each
compound, 1 mL of a 10
-4
M sol uti on i n methanol was added
to 2 mL of DPPH sol uti on (10 mg/L). Fi ve mi nutes l ater, the
absorbance was measured at 517 nm. A reference sampl e was
prepared wi th 1 mL of methanol . The anti radi cal acti vi ty was
cal cul ated as a percentage of DPPH decol orati on usi ng the
fol l owi ng equati on:
Determination of the Rate of Flavonoid Oxidation.
The spectra of fl avonol s exhi bi t two major absorpti on peaks,
whi ch are commonl y referred to as band I (350-385 nm) and
band I I (240-280 nm). The UV absorbance of band I of the
fl avonoi d spectra was measured at the begi nni ng and after 2
h of keepi ng of the fl avonoi d sol uti on i n the same condi ti ons
as the anti oxi dant acti vi ty determi nati on, but wi thout addi ti on
of -carotene.
* Author to whom correspondence shoul d be addressed
(tel ephone 048 81 886 3421, ext. 205; fax 048 81 886 4547;
e-mai l burda@i ung.pul awy.pl ).
% anti oxi dant acti vi ty ) 100
[1 - (A
s
0
- A
s
120
)/(A
c
0
- A
c
120
)]
anti radi cal acti vi ty ) 100
(1 - absorbance of sampl e/absorbance of reference)
2774 J. Agric. Food Chem. 2001, 49, 27742779
10.1021/jf001413m CCC: $20.00 2001 American Chemical Society
Published on Web 05/30/2001
Water/Chloroform Partition Coefficient Determina-
tion. The coeffi ci ent Kw/c was cal cul ated as the rati o of the
absorbance of a water sol uti on of an i ndi vi dual compound,
measured at the maxi mum of absorpti on, before and after
extracti on wi th chl oroform at 50 C.
RESULTS AND DI SCUSSI ON
The hi ghest anti oxi dant acti vi ti es were establ i shed
for the syntheti c anti oxi dant 2,6-bi s(1,1-di methyl ethyl -
4-methyl phenol ) (BHT), D,L-R-tocopherol , and i ts acetate
(Tabl e 1; Fi gures 1 and 2). Sl i ghtl y l ower, but sti l l hi gh,
anti oxi dant acti vi ty was shown by a homogeneous group
that i ncl uded fi seti n, kaempferol , gal angi n, querceti n,
robi neti n, mori n, and kaempferi de. Al l of these com-
pounds are fl avonol s wi th a free hydroxyl group at the
C-3 posi ti on. Thi s suggested that the fl avonol C-3
hydroxyl group i s responsi bl e for the hi gh i nhi bi ti on of
-carotene oxi dati on i n the heterogeneous system. Com-
pari son of the anti oxi dant acti vi ty of fl avonol agl ycons
wi th the acti vi ty of thei r gl ycosi des or methyl deri va-
ti ves showed that the bl ockage of the C-3 hydroxyl group
resul ted i n a total l oss of anti oxi dant acti vi ty (Tabl e 2).
Gl ycosyl ati on or methyl ati on of other fl avonol hydroxyl s
di d not produce such an effect.
These resul ts are i n agreement wi th those presented
by Chung et al . (18) for fl avonoi ds of Chorizantediffusa
i n whi ch the fl avonol wi th a free 3-OH group had a
hi gher anti oxi dant potenti al than i ts substi tuted deri va-
ti ves. Joyeux et al . (8) al so showed the hi ghest anti l i -
poperoxi dant acti vi ty for free fl avonol s; taxi fol i n (di hy-
drofl avonol ), ruti n, and fl avones had di sti nctl y l ower
acti vi ty.
Some apparent di screpanci es wi th the concl usi on that
the abi l i ty of fl avonoi ds to i nhi bi t oxi dati on processes
i s control l ed by the presence of the doubl e bond between
C-2 and C-3 and a free hydroxyl i n C-3 posi ti on were
found i n the present study. The anti oxi dant acti vi ti es
obtai ned for 3,5,7,3,4-pentahydroxy-5-methoxyfl avone
(l ari cytri n), l ari cytri n 3-O-gl ucosi de, and 3,5,7,3,4,5-
pentahydroxyfl avone (myri ceti n) were l ow. The l ower
than expected anti oxi dant acti vi ty val ues obtai ned for
these compounds can be expl ai ned by thei r hi gh sensi -
ti vi ty to oxi dati on, whi ch causes thei r rapi d oxi dati on
and parti al decomposi ti on duri ng measurement. Oxi da-
ti on was al so observed for the other fl avonol s but to a
l esser extent (Tabl e 3).
The fl avonoi ds exami ned showed di fferent sol ubi l i ty
patterns i n chl oroform. As suggested previ ousl y, these
di fferences i n sol ubi l i ty may i nfl uence resul ts of tests
(19), but i n the present experi ments the anti oxi dant
acti vi ty val ues obtai ned di d not correl ate wi th water-
chl oroform parti ti on coeffi ci ents. Thi s may suggest that
parti ti on of the compounds between two phases di d not
si gni fi cantl y i nfl uence oxi dati on resul ts.
As shown by Russo and co-workers (20), on the basi s
of semi empi ri cal cal cul ati ons, radi cal s formed by H

removal from hydroxyl s at C-3 and C-4 may be i nvol ved

i n the anti oxi dant properti es of querceti n. The resul ts
Table 1. Antioxidant Activity of Flavonoids (3.9 10
-6
M)
a
in an Aqueous Emulsion of Linoleic Acid/-Carotene
at 50 C and Water/ChloroformPartition Coefficient for
Flavonoids at 50 C
compound
anti oxi dant
acti vi ty (%)
parti ti on
coeffi ci ent
(Kw/c)
BHT 95.3
a
nd
D,L-R-tocopherol 95.8
a
nd
D,L-R-tocopherol acetate 88.6
a
nd
kaempferol (1) 65.3
b
0.52
gal angi n (2) 64.9
b
0.04
querceti n (3) 63.6
b
5.48
mori n (18) 63.5
b
10.39
robi neti n (5) 61.7
b
15.44
fi seti n (6) 61.6
b
9.02
kaempferi de (7) 60.0
b
0.07
3-hydroxyfl avone (8) 59.4
b
0.18
coumestrol (42) 38.7
c
0.34
l ari cytri n (9) 28.5
d
0.84
l ari cytri n 3-O-gl ucosi de (10) 26.2
d,e
2.36
myri ceti n (11) 18.4
e
6.69
hespereti n (31) 4.7
f
0.02
3,5,7,3,4,5-hexamethoxyfl avone (12) 2.6
e,f
0.05
3,5,7,3,4-pentamethoxyfl avone (13) 1.1
f,g
nd
l ari cytri n 3,3-O-di gl ucosi de (14) 1.1
f,g
1.07
7-hydroxyfl avone(21) 0.0
f,g
0.08
fl avone (19) -1.5
f,g,h
0.01
5-hydroxyfl avone (20) -4.0
f,g,h
0.14
querceti n 3-O-gl ucosi de-7-O-
rhamnosi de (15)
-6.2
g,h
92.44
l ari cytri n 3,7,3-O-tri gl ucosi de (16) -6.2
g,h
16.32
ruti n (17) -10.2
h,i
113.47
taxi fol i n (di hydroquerceti n) (33) -16.8
i ,j
21.31
nari ngeni n (29) -16.8
i ,j
0.55
GB-1a (bi fl avanone) (35) -16.9
i ,j
nd
kaempferol 3,7-O-di rhamnosi de (4) -17.5
i ,j
48.58
formononeti n (39) -20.4
j,k
0.04
bi ochani n A (41) -20.4
j,k
0.06
chrysi n (22) -20.8
j,k,l
0.08
fl avanone (28) -23.0
j,k,l
0.16
fusti n (di hydrofi seti n) (32) -23.4
j,k,l
13.21
geni stei n (40) -24.6
j,k,l ,m
0.75
l uteol i n 7-O-gl ucosi de (27) -25.3
j,k,l ,m
63.86
8-methoxyfl avone (23) -29.2
k,l ,m
0.01
api geni n 8-C-gl ucosi de (vi texi n) (25) -29.6
k,l ,m
nd
GB-1 (bi fl avanone) (34) -30.1
l ,m
nd
dai dzei n (38) 32.9
m
7.16
nari ngi n (30) 47.4
n
55.90
api geni n 7-O-gl ucosi de (26) -63.9
o
10.13
api geni n (24) -78.8
p
1.21
a
The concentrati on i s cal cul ated on the basi s of the total vol ume
of the emul si on. Means fol l owed by the same l etter are not
si gni fi cantl y di fferent by LSD mul ti pl e-range test at 5%l evel . For
fl avonoi d structure see Fi gures 1 and 2. nd, not determi ned.
Table 2. Effect of Glycosylation or Methylation of Flavonols on Their Antioxidant Activity
a
fl avonol (agl ycon)
anti oxi dant
acti vi ty (%) gl ucosi de or methoxyl deri vati ve
anti oxi dant
acti vi ty (%)
kaempferol (1) 65.3 kaempferi de (7) 60.0
kaempferol (1) 65.3 kaempferol 3,7-O-di rhamnosi de (4) -17.5
querceti n (3) 63.6 querceti n 3-O-gl ucosi de-7-O-rhamnosi de (15) -6.2
querceti n (3) 63.6 querceti n 3-O-rhamnogl ucosi de (ruti n) (17) -10.2
querceti n (3) 63.6 3,5,7,3,4-pentamethoxyfl avone (13) 1.1
l ari cytri n (9) 28.5 l ari cytri n 3-O-gl ucosi de (10) 26.2
l ari cytri n (9) 28.5 l ari cytri n 3,3-O-di gl ucosi de (14) 1.1
l ari cytri n (9) 28.5 l ari cytri n 3,7,3-O-tri gl ucosi de (16) -6.2
l ari cytri n (9) 28.5 3,5,7,3,4,5-hexamethoxyfl avone (12) 2.6
myri ceti n (11) 18.4 3,5,7,3,4,5-hexamethoxyfl avone (12) 2.6
a
For fl avonoi d structures see Fi gure 1.
Flavonoid Antioxidant Activity J. Agric. Food Chem., Vol. 49, No. 6, 2001 2775
of the present experi ments reveal that i n the heteropha-
si c model i ng system used the abi l i ty to i nhi bi t -caro-
tene oxi dati on by fl avonoi ds depends pri mari l y on the
free hydroxyl at C-3 and the doubl e bond between C-2
and C-3.
Of the remai ni ng fl avonoi ds of di fferent structures
onl y coumestrol had moderate anti oxi dant acti vi ty
(38.7%). Other fl avonoi ds showed no anti oxi dant pro-
perti es under these experi mental condi ti ons. I t shoul d
be noted that there were numerous fl avonoi ds that,
i nstead of i nhi bi ti ng oxi dati on, were abl e to enhance thi s
process, an aspect that requi res further el uci dati on.
Fl avonol 3-O-gl ycosi des have a bl ocked C-3 hydroxyl
group, whi ch i n l i ne wi th the theory presented here,
resul ts i n thei r i nabi l i ty to i nhi bi t oxi dati on. The
anti oxi dant acti vi ty for these compounds fal l s wi thi n the
range of 1 to -20%, whi ch cl assi fi es them as total l y
i nacti ve. However, i n bi ol ogi cal systems gl ycosi des may
undergo enzymati c hydrol ysi s, resul ti ng i n the forma-
ti on of acti ve agl ycons (21). For exampl e, ruti n (quer-
ceti n 3-O-rhamnogl ucosi de), whi ch i s commonl y found
i n pl ants, upon the hydrol ysi s of the gl ycosi de bond
produces querceti n, a hi ghl y anti oxi dati ve agl ycon.
Tabl e 4 show resul ts obtai ned for the DPPH decol -
orati on test. The fl avonoi ds wi thout any hydroxyl group
(fl avone, fl avanone, and 8-methoxyfl avone) or wi th the
free hydroxyl s onl y at C-5 and/or at C-7 (5-hydroxyfl a-
vone, 7-hydroxyfl avone, and 5,7-di hydroxyfl avone) had
no effect on scavengi ng of free radi cal s. Al l fl avonol s
wi th a free hydroxyl i n the C-3 posi ti on, whi ch were very
effecti ve anti oxi dants, al so had hi gh abi l i ty to scavenge
DPPH radi cal s. I f the compounds that are very sensi ti ve
to oxi dati on (myri ceti n, l ari cytri n, and i ts 3-O-gl ucosi de)
are i gnored, the correl ati on between anti oxi dant acti vi ty
and anti radi cal acti vi ty i s very strong (r ) 0.92) for thi s
group of fl avonoi ds.
The remai ni ng fl avonoi ds exami ned showed anti radi -
cal acti vi ty wi thi n a wi de range from a few percent to
>90%. Al l of these compounds had a free hydroxyl group
at the C-4 posi ti on. I t appears that the presence of thi s
hydroxyl group i s essenti al for the anti radi cal acti vi ty
of thi s group of fl avonoi ds. The anti radi cal effecti veness
Table 3. Rate of Oxidation of Selected Flavonoids Measured by the Decrease of Absorption in Band I
a
compound
anti oxi dant
acti vi ty (%)
absorpti on decl i ne i n
band I of fl avonoi d spectra (%)
3,5,7-tri hydroxyfl avone
b
(2) 64.9 20.6
3,7,3,4-tetrahydroxyfl avone
c
(6) 67.2 23.0
3,5,7,4-tetrahydroxyfl avone
d
(1) 65.3 14.9
3,5,7,3,4-pentahydroxyfl avone
e
(3) 63.6 20.7
3,5,7,5,4-pentahydroxy-3-methoxyfl avone
f
(9) 28.5 46.5
3,5,7,3,4,5-hexahydroxyfl avone
g
(11) 18.4 42.7
a
Band I absorpti on peak at 350-385 nm. For fl avonoi d structures see Fi gure 1.
b
Tri vi al name: gal angi n.
c
Tri vi ai l name: fi seti n.
d
Tri vi al name: kaempferol .
e
Tri vi al name: querceti n.
f
Tri vi al name: l ari ci tri n.
g
Tri vi al name: myri ceti n.
Table 4. Antiradical Activities of Flavones, Flavanones, and Biflavanones (3.3 10
-5
M) in a Methanol Solution of
DPPH (1.6 10
-5
M)
a
compound
anti radi cal
acti vi ty (%)
doubl e
bond C2-C3 C3-OH C4-OH
o-di -OH
B-ri ng
mori n (18) 96.5
a
+ + + +
taxi fol i n (di hydroquerceti n) (33) 94.8
a,b
+ + +
kaempferol (1) 93.5
b,c
+ + +
GB-2 (bi fl avanone) (36) 92.8
b,c
+ + +
fusti n (di hydrofi seti n) (32) 91.9
c,d
+ + +
gal angi n (2) 91.8
c,d
+ +
ruti n (17) 90.9
c,d
+ + +
querceti n (3) 89.8
d,e
+ + + +
l uteol i n 7-O-gl ucosi de (27) 87.6
e,f
+ + +
querceti n 3-O-gl ucosi de-7-O-rhamnosi de (15) 86.8
f,g
+ + +
l ari cytri n (9) 84.6
g,h
+ + + +
l ari cytri n 3-O-gl ucosi de (10) 83.8
h
+ + +
robi neti n (5) 82.3
h
+ + + +
fi seti n (6) 79.0
i
+ + + +
myri ceti n (11) 72.8
j
+ + + +
kaempferol 3,7-di rhamnosi de (4) 70.6
j
+ +
3-hydroxyfl avone (8) 66.0
k
+ +
api geni n 7-O-gl ucosi de (26) 34.8
l
+ +
hespereti n (31) 30.0
l
+
vi texi n (25) 21.0
m
+ +
3,5,7,3,4,5-hexamethoxyfl avone (12) 12.6 +
GB-1a (bi fl avanone) (35) 11.2
n,o
+
GB-1 (bi fl avanone) (34) 9.5
o
+ +
nari ngeni n (29) 6.3
p
+
GB-2a (bi fl avanone) (37) 5.6
p
+
nari ngi n (30) 4.7
p,r
+
7-hydroxyfl avone (21) 2.8
r,s
+
fl avanone (28) 2.6
r,s
fl avone (19) 1.5
s
+
chrysi n (22) 1.1
s
+
api geni n (24) 0.7
s
+ +
8-methoxyfl avone (23) 0.7
s
+
5-hydroxyfl avone (20) 0.6
s
+
a
Means fol l owed by the same l etter are not si gni fi cantl y di fferent by LSD mul ti pl e-range test at the 5% l evel ; see Fi gures 1 and 2 for
fl avonoi d structure.
2776 J. Agric. Food Chem., Vol. 49, No. 6, 2001 Burda and Oleszek
of thi s group can be strengthened by certai n other
structural features of the fl avonoi d mol ecul e such as a
doubl e bond i n the C-ri ng (C2-C3) and hydroxyl groups
at C-3 and/or C-3 posi ti ons. Fl avanones wi th a si ngl e
bond at C2-C3, whi ch have onl y one hydroxyl group i n
the B-ri ng at C-4, showed very l ow anti radi cal acti vi ty
(nari ngi n and nari ngeni n). Fl avones and fl avonol s wi th
a substi tuted hydroxyl group at the C-3 posi ti on, whi ch
have onl y a C-4 hydroxyl i n the B-ri ng, showed di s-
ti nctl y hi gher acti vi ty, rangi ng from 20 to 70%. Onl y
api geni n showed no anti radi cal properti es. Strong an-
ti radi cal acti vi ty, comparabl e to that expressed by free
fl avonol s, was shown by compounds wi th an o-di hydroxy
system i n the B-ri ng. Thi s group i ncl udes di hydro-
fl avonol s, fl avonol 3-O-gl ycosi des, and fl avones. I t ap-
pears that the reason for thei r hi gh acti vi ty i s a strong
effect of the C-3 hydroxyl on the reacti vi ty of the
hydroxyl at C-4. Thi s i s i n agreement wi th concl usi ons
that the o-di hydroxy system i n the B-ri ng of fl avonoi ds
i s hi ghl y effecti ve agai nst free radi cal s (8). We were
Figure 1. Structures of fl avonol s, fl avones, fl avanones, and di hydrovl avonol s tested.
Flavonoid Antioxidant Activity J. Agric. Food Chem., Vol. 49, No. 6, 2001 2777
unabl e to expl ai n the i nfl uence of the methoxyl groups
on anti radi cal properti es. The sl i ght acti vi ty found
for hespereti n (5,7,3-tri hydroxy-4-methoxyfl avanone)
suggested that a methoxyl substi tuted i n certai n
posi ti ons can i ncrease the anti radi cal acti vi ty of fl a-
vonoi ds.
The anti radi cal acti vi ty of bi fl avanones can be ex-
pl ai ned by the same factors. From the four compounds
exami ned, onl y GB-2, whi ch consi sts of nari ngeni n and
di hydroquerceti n moi eti es, had the same acti vi ty as
di hydroquerceti n i tsel f. The other bi fl avanones, whi ch
compri sed onl y i nacti ve monomers, showed very l ow
acti vi ty. Three i sofl avones exami ned showed very l ow
anti radi cal acti vi ty. We have i nsuffi ci ent resul ts to
di scuss any structure-acti vi ty rel ati onshi p for thi s
fl avonoi d group. The moderate anti radi cal acti vi ty of
coumestrol correl ated wi th the anti oxi dant acti vi ty of
thi s compound.
ABBREVI ATI ONS USED
BHT,2,6-bis(1,1-dimethylethyl-4-methylphenol);DPPH,
1,1-di phenyl -2-pi cryl hydrazyl .
ACKNOWLEDGMENT
We thank Prof. Mari an Jurzysta for provi di ng 15
sampl es of fl avonoi ds used i n thi s study.
LI TERATURE CI TED
(1) Ri ce-Evans, C. Screeni ng of phenol i cs and fl avonoi ds for
anti oxi dant acti vi ty. I n Antioxidant Food Supplements
in Human Health; Packer, L., Hi ramatsu, M., Yoshi ka-
wa, T., Eds.; Academi c Press: San Di ego, CA, 1999; pp
239-253.
(2) Jovanovi c, S. V.; Steenken, S.; Tosi c, M.; Marjanovi tz,
B.; Si mi c, M. G. Fl avonoi ds as anti oxi dants. J . Am.
Chem. Soc. 1994, 116, 4846-4851.
(3) Jovanovi c, S.; Steenken, S.; Si mi c, M.; Hara, Y. Anti -
oxi dant properti es of fl avonoi ds: Reducti on potenti al s
and el ectron-transfer reacti ons of fl avonoi d radi cal s. I n
Flavonoids in Health and Disease; Ri ce-Evans, C.,
Packer, L., Eds.; Dekker: New York, 1998; pp 137-161.
(4) Bors, W.; Hel l er, W.; Mi chel , C.; Saran, M. Fl avonoi ds
as anti oxi dants: determi nati on of radi cal -scavengi ng
effi ci enci es. Methods Enzymol. 1990, 186, 343-355.
(5) Bors, W.; Mi chel , C.; Stettmai er, K. Anti oxi dant effects
of fl avonoi ds. Biofactors 1997, 6, 399-402.
(6) Cao, G.; Sofi c, E.; Pri or, R. L. Anti oxi dant and prooxi -
dant behavi our of fl avonoi ds: Structure-acti vi ty rel a-
ti onshi ps. FreeRadical Biol. Med. 1977, 22, 749-760.
(7) Foti , M.; Pi attel l i , M.; Baratta, M. T.; Ruberto, G.
Fl avonoi ds, coumari ns, and ci nnami c aci ds as anti oxi -
dants i n a mi cel l ar system. Structure-acti vi ty rel ati on-
shi p. J . Agric. Food Chem. 1996, 44, 497-501.
(8) Joyeux, M.; Lobstei n, A.; Anton, R.; Morti er, F. Com-
parati ve anti l i poperoxi dant, anti necroti c and scavengi ng
properti es of terpenes and bi fl avones from Ginkgo and
some fl avonoi ds. Planta Med. 1995, 61, 126-129.
(9) Burda, S.; Jurzysta, M. I sol ati on and i denti fi cati on of
fl avonoi ds from Medicagolupulina L. fl owers. Acta Soc.
Bot. Pol. 1988, 57, 563-571.
(10) Di etri ch-Szostak, D.; Ol eszek, W. Effect of processi ng
on the fl avonoi d content i n buckwheat (Fagopyrum
esculentumMoench) grai n. J . Agric. Food Chem. 1999,
47, 4384-4387.
(11) Burda, S.; Ol eszek, W.; I gi l e, G. O. Chromatography of
bi fl avanones from Garcinia kola frui t. Proceedings,
Chromatography Seminar Science-I ndustry; Lubl i n,
Pol and, 1996; p 9.
(12) Gorski , P. M.; Jurzysta, M.; Rzadkowska-Bodal ska, H.
Fl avonoi ds from the bi rds trefoi l seeds (Lotus cornicu-
latus L.). Acta Soc. Bot. Pol. 1975, 44, 289-295.
(13) Gorski , P. M.; Burda, S.; Jurzysta, M.; Pl oszynski , M.
A method of i sofl avones i sol ati on from red cl over to be
used as standards for determi nati on. Acta Agrobot.
1989, 42, 191-196.
(14) I gi l e, G. O.; Ol eszek, W.; Jurzysta, M.; Burda, S.;
Fafunso, M.; Fasanmade, A. A. Fl avonoi ds from Ver-
nonia amygdalina and thei r anti oxi dant acti vi ti es. J .
Agric. Food Chem. 1994, 42, 2445-2448.
(15) Ol eszek, W.; Stochmal , A.; Karol ewski , P.; Si monet, A.
M.; Maci as, F. Fl avonoi ds from needl es of Pinus silves-
tris as a markers of chemi cal bi ogenesi s of pi ne. Pro-
ceedings, Conferenceon Biochemical Responses in En-
vironmental I nteractions; I UNG, Pul awy, Pol and, 2000;
p 83.
(16) Pratt, D. E. Natural anti oxi dants from pl ant materi al .
I n Phenolic Compounds in Food and Their Effect on
Health I I : Antioxidants and Cancer Prevention; Huang,
M.-T., Ho, C.-T., Lee, C. Y., Eds.; Ameri can Chemi cal
Soci ety: Washi ngton, DC, 1992; pp 54-71.
Figure 2. Str uctur es of bi fl avanones, i sofl avones, and
coumestrol tested.
2778 J. Agric. Food Chem., Vol. 49, No. 6, 2001 Burda and Oleszek
(17) Lee, S. K.; Mbwambo, Z. H.; Chung, H.-S.; Luyengi , L.;
Games, E. J. C.; Mehta, R. G.; Ki nghorn, A. D.; Pezzuto,
J. M. Eval uati on of the anti oxi dant potenti al of natural
products. Combin. Chem. High Throughput Screen.
1998, 1, 35-46.
(18) Chung, H. S.; Chang, L. C.; Lee, S. K.; Shamon, L. A.;
van Breemen, R. B.; Metha, R. G.; Farnsworth, N. R.;
Pezzuto, J. M.; Ki nghorn, A. D. Fl avonoi d consti tuents
of Chorizanthediffusa wi th potenti al cancer chemopre-
venti ve acti vi ty. J . Agric. Food Chem. 1999, 47, 36-41.
(19) Huang, S.-W.; Hopi a, A.; Schwarz, K.; Frankel , E. N.;
German, J. B. Anti oxi dant acti vi ty of R-tocopherol and
trol ox i n di fferent l i pi d substrates: bul k oi l s vs oi l -i n-
water emul si ons. J . Agric. Food Chem. 1996, 44, 444-
452.
(20) Russo, N.; Toscano, M.; Uccel l a, N. Semi empi ri cal
mol ecul ar model i ng i nto querceti n reacti ve si te: struc-
tural , conformati onal , and el ectroni c features. J . Agric.
Food Chem. 2000, 48, 3232-3237.
(21) Terao, J. Di etary fl avonoi ds as pl asma anti oxi dants on
l i pi d peroxi dati on: Si gni fi cance of metabol i c conversi on.
I n Antioxidant Food Supplements in Human Health;
Packer, L., Hi ramatsu, M., Yoshi kawa, T., Eds.; Aca-
demi c Press: San Di ego, CA, 1999; pp 255-268.
Recei ved for revi ew November 27, 2000. Revi sed manuscri pt
recei ved March 28, 2001. Accepted March 29, 2001.
JF001413M
Flavonoid Antioxidant Activity J. Agric. Food Chem., Vol. 49, No. 6, 2001 2779