Food and Chemical Toxicology 42 (2004) 659–666

www.elsevier.com/locate/foodchemtox

Antimutagenic, antioxidant and free radical scavenging activity of

ethyl acetate extracts from white, yellow and red onions
Mi-Yae Shona, Sang-Do Choib, Goon-Gjung Kahngb, Sang-Hae Namb, Nak-Ju Sunga,*
a
Department of Food and Nutrition, Institute of Agriculture & Life Science, Gyeongsang National University, 900 Gaza-dong chinju city,
South Korea
b
Department of Food Processing, Jinju National University, 660-758, Chilam-dong chin-ju city, South Korea

Received 31 May 2003; received in revised form 15 November 2003; accepted 8 December 2003

Abstract
The beneficial effects of red, yellow and white onion extracts have been assessed by antioxidant activity and antimutagenic
activity. And the effects compared to BHT and ascorbic acid. Total phenolic compounds and flavonoids in onion extracts were
determined. Yellow onion extract had more organic acid and free sugar than those detected in the white and red onion extract. The
scavenging activity of DPPH radical and H2O2 were increased depending on the concentration. The antioxidant activities using b-
carotene-linoleate system and reducing power were increased but the effect was small to that of BHT and ascorbic acid. After
digested, extracts showed antimutagenic activities, and it seems that they inhibit the mutagenicity for digesting. This study
demonstrated that the antimutagenicities and antioxidant properties of ethyl acetate extract against mutagens were related to their
phenols and flavonoids, which are heat stable and losses digestive juices are relatively low.
# 2003 Elsevier Ltd. All rights reserved.
Keywords: Antioxidant activity; Antimutagenic activity; Phenols; Flavonoids; Onion extracts; Digestive solutions

1. Introduction This protective effect is attributed to flavonoids and

Flavonoids and phenolic compounds are occurring in
food plants and are a common component in the human
diet. Food-derived flavonoids and phenolic compounds
such as the flavonols quercetin, kaempferol, gallic acid
and myricetin have been reported to exhibit a wide
range of biological effects, including antibacterial, anti-
viral, anti-inflammatory, antiallergic actions (Huang et
al., 1983). In addition, flavonoids inhibit lipid peroxi-
dation and exert these effects as antioxidants, free radi-
cal scavengers, and chelators of divalent cations.
Epidemiological research on the relation between flavo-
noid intake and disease risk in humans is needed to
support the findings of these experimental studies. Epi-
demiological studies investigating relations between diet
and disease, a protective effect of the consumption of
vegetables and fruits on various forms has been studied
(Steinmetz and Potter, 1991).
* Corresponding author. Tel.: +82-55-751-5975; fax: +82-55-751-
5974.
E-mail address: nuruksmy@hanmail.net (N.-J. Sung).
0278-6915/$ - see front matter # 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/j.fct.2003.12.002
phenolic compounds present in these foods. To investi-
gate the mechanisms of these effects in animal bodies,
several studies have been conducted to determine their
absorption and distribution. Onions have been shown
to contain large amounts of flavonoids, and constitute
one of the major sources of flavonoids in diets (Knekt et
al., 1996). Onion, a vegetable member of the genus
Allium, has been reported to promote cardiovascular
health exhibiting decreased rates of atherosclerosis or
thrombotic disease in populations with increased onion
intake (Kendler, 1987). These beneficial effects of onion
have been attributed to its ability to inhibiot platelet
aggregation and thromboxane formation (Srivastava,
1986). However there is little chemopreventive inform-
ation on effects of extracts of three varieties of onions.
This research is needed to investigate the character the
components in onions.
Flavonoids are low molecular weight polyphenolic
substances based on the flavan nucleus. The biochemical
activities of flavonoids and their metabolites depend on
their chemical structure and the relative orientation of
various moieties on the molecule. They are hydrolyzed
660 M.-Y. Shon et al. / Food and Chemical Toxicology 42 (2004) 659–666
by intestinal flora to produce the biologically active
aglycone (sugar-free flavonoid). There is now increasing
evidence about absorption of flavonoids and some of
them have been detected in human plasma and other
biological fluids (Bourne and Rice-Evans, 1999; Pietta
& Simonetti, 1999; Terao, 1999). In the human study,
the absorption of orally administered quercetin was
24%, while the absorption of quercetin glycosides from
onions was 52% (Hollman et al., 1995). Dietary inhibi-
tors of mutagenesis and carcinogenesis are of particular
interest because they may be useful for human cancer
prevention on recently, several flavonoids and phenolic
compounds have been demonstrated to have an anti-
mutagenic effect on various mutagens or carcinogens
(Francis et al., 1989; Chang-qi et al., 1994; Birt et al.,
1986). Also dietary flavonoids, a group of polyphenols,
suppressed the metabolism of the widespread food car-
cinogen 2-amino-3-methylimidazo-[4,5-f]quinoline(IQ)
(Edenharder et al., 1997), in the presence of a hepatic
activation system derived from Aroclor 1254-treated
rats, is a typical heterocyclic amine mutagen found in
cooked foods. The ellagic acid scavenged the reactive
intermediates of polycyclic aromatic hydrocarbons
(Sayer et al., 1982). Polyphenols can also impair the
promotion stage of carcinogenesis as a result of their
antioxidant activity (Vinson et al., 1995).
Plant flavonoids are known to be easily degraded in
alkaline solutions (Tamura and Yoshino, 1997). Human
intestinal and pancreatic juices also are known to be
mildly alkaline, with pH values of 8.3 and 7.0 to 8.5,
respectively. Therefore, in this study, we determined the
antimutagenic activity and antioxidant activity with
onion extracts. Furthermore, we also investigated the
antimutagenic activity of onion extracts after digestion
in simulated gastric and intestinal juice under various
conditions with mutagen. The aim of the present study
is a need to know the absorption of flavonoids and
phenolic acids whether they are loss or not in the simu-
lated gastric and intestinal juices. If they will absorb and
metabolize, they have been shown to inhibit about
mutagen.
2. Materials and methods
2.1. Materials
All solvents used were of HPLC grade, Reagents and
chemicals were purchased from Sigma-Aldrich, Wako
and Fluka. b-carotene, linoleic acid, polyoxyethylene
sorbitan monopalmitate (Tween 40), buthylated hydro-
xytoluene (BHT), l-ascorbic acid, pepsin(from hog sto-
mach), pancreatin(from porcine pancreas), 2,2-
diphenyl-picrylhydrazyl(DPPH), liloeic acid, EDTA,
ferric chloride, hydrogenperoxide (H2O2, 30%, v/v),
vanillin, potassium ferricyanide, DMSO and MNNG
were purchased from Sigma Chemical Co. (St. Louis,
Mo, USA). IQ was purchased from Wako Chemical
Inc. The Salmonella typhimurium strains TA98 and
TA100 were purchased from KCTC (Korean Collection
for Type Cultures). Aroclor 1254-induced hepatic S9
was made for the activation system in the case of IQ.
Agar and Nutrient Broth No.2 were purchased from the
Difco Laboratories (Detroit, USA) and Oxoid (Hamp-
shire, UK), respectively.
2.2. Preparation of onion extracts
The onions varieties white skinned (Albion), yellow
skinned (Rijnsburger) and red skinned (varity Red
Baron) were purchased from local markets. Each onion
was skinned, chopped and lyophilized. The lyophilized
onions were ground to a fine power. Ground onion
powers (50 g) were extracted with 85% ethyl acetate
room temperature for 12 h (3 times with 500 ml),
respectively. Ethyl acetate extract contain hydrophilic
and lipophilic nature. It is of general interest to measure
the total antioxidant activity. The extracts were con-
centrated in a vacuum evaporator (EYELA, Japan) at
below 40
C. All of the concentration were lyophilized
and stored at 20
C.
2.3. Determination of phenolic compounds
The contents of total phenols were analyzed by the
method of Folin and Denis (Folin and Denis, 1915),
reading samples on a UV/Vis spectrophotometer at 760
nm. The each extract was mixed with 5 ml of Folin–
Denis phenol reagent, 10 ml of Na2CO3 and diluted by
a factor 100 with distilled water. The total phenol con-
tent of each solvent extract was estimated by compar-
ison with a standard curve generated from analysis of
caffeic acid.
The content of total flavonoids was analyzed color-
imetrically by the Davis method using myricetin as a
standard (AOAC, 1995). The each extract (0.01 g) was
mixed 30 ml of distilled water and 30 ml of methanol.
After boiling for 30 min in 90
C, cooled and adjusted
100 ml of distilled water. Diluted solution was filtered. 1
ml of filtrared solution was mixed with 10 ml of diethy-
lene glycol and 1 ml of 1N-NaOH. The absorbances of
sample and blank were measured at 420 nm after stand
for 30 min in the dark at room temperature.
Organic acid was determined with HPLC analysis
(AOAC, 1995). The each extract (1 g) was dissolved in
distilled water (50 ml) and filtered through 0.45 mm fil-
ter. The filtrate was stirred for 30 min. The filtrate (0.2
ml) was injected into the HPLC apparatus.
Chromatography was performed on a m-Bondapack
C-18 column (Water-Millipore Corporation. Milford,
MA, USA). The mobile phase was water/methanol
10:80 containing 2% of acetic acid. The flow-rate was
 M.-Y. Shon et al. / Food and Chemical Toxicology 42 (2004) 659–666
set at 1.0 ml/min. Detection was effected at 330 nm. The
peak assignments were based on the retention times of
the single compound. Calibration graphs were plotted
showing a linear relationship between concentrations
versus peak-areas for reference compound.
Free sugar was determined with HPLC analysis
(AOAC, 1995). The extract (1 g) was dissolved in dis-
tilled water (30 ml) and centrifuged at 8000 rpm for 20
min. The supernatant make up 50 ml with distilled
water and filtered through 0.2 mm filter. The filtrate was
passed with sep-pak C18 and injected into the HPLC
apparatus (Water-Millipore Corporation. Milford, MA,
USA). Chromatography was performed carbohydrate
column. The mobile phase was 80% acetonitrile. The
flow-rate was set at 2.0 ml/min. The peak assignments
were based on the retention times of the single
compound.
2.4. DPPH radicals scavenging activity
The scavenging activity of onion extracts by DPPH
radicals was measure according to the method of Shimada
et al. (1992). Ethanolic solutions of DPPH (104) and
onion extracts solutions were mixed so that the final
mass ratios were extracts: DPPH.=5.5:1 and reference
compounds: DPPH.=0.5:1. The samples were incu-
bated for 15 min in the dark was at 30
C and the
decrease in absorbance at 517 nm was measured against
ethanol using a spectrophotometer. Ethanol was used to
zero the spectrophotometer; a blank sample containging
the same amount of ethanol and DPPH. was prepared
and measured daily, stored in a flask covered and kept
in the dark at 4
C between the measurements. All
determinations were performed in triplicate.
2.5. Antioxidant assay using
-carotene linoleate model
system
The antioxidant activity of extracts was evaluated by
the b-carotene linoleate model system (Miller, 1971). A
solution of b-carotene was prepared by dissolving 2 mg
of b-carotene in 10 ml of chloroform. Two mililitres of
this solution were pipetted into a 100 ml round bottom
flask. After chloroform was removed under vacuum, 40
mg of purified linoleic acid, 400mg of Tween 40 emulsi-
fier, and 100 ml of aerated distilled water were added to
the flask with vigorous shaking. Aliquots (4.8 ml) of this
emulsion were transferred into a different test tubes
containing different concentration of the extract. BHA
was used for comparative purposes. As soon as the
emulsion was added to each tube, the zero time absor-
bance was measured at 470 nm using a spectro-
photometer (CE2021, CECIL, England). The tubes
were placed at 50
C in a water bath. Measurement of
absorbance was continued until the colour of b-carotene
disappeared for the blank, devoid of b-carotene, were
661
prepared for back ground subtraction. Antioxidant
activity (AA) was calculated using the following
equation:
AA ¼ b-carotene content after 2 h of assay=


initial b-carotene content  100
2.6. Reducing power activity
The reducing power of extracts was determined by the
method of Yen and Duh (1993). Different concentra-
tions of extracts were mixed with 2.5 ml of phosphate
buffer (200 mM, pH 6.6) and 2.5 ml of 1% potassium
ferricyanide. The mixtures were incubated for 20 min at
50
C. After incubation, 2.5 ml of 10% trichloroacetic
acid were added to the mixtures, followed by cen-
trifugation at 650g for 10 min. The upper layer (5 ml)
was mixed with 5 ml of distilled water and 1 ml of 0.1%
ferric chloride and the absorbance of the resultant solu-
tion was measured at 700 nm.
2.7. Hydrogen peroxide scavenging activity
The extract was dissolved in 3, 4 ml of 0.1 M phos-
phate buffer(pH 7.4) and mixed with 600 ml of 43 mM
solution of hydrogen peroxide. The absorbance value
(at 230 nm) of the reaction mixture was recorded from 0
min to 40 min and then at every 10 min. For each con-
centration, a separate blank sample was used for back
ground subtraction (Ruch et al., 1989).
2.8. Preparation of the gastric and intestinal juice
The simulated gastric and intestinal juice was pre-
pared as follows (USP 23).
The gastric fluid consisted of 2.0 g of NaCl and 3.2 g
of Pepsin, dissolved in 7.0 ml of HCl, And water was
added to make 1000 ml. The pH of the gastric fluid was
pH 1.2. The intestinal fluid consisted of 6.8 g of
KH2PO4, dissolved in 250 ml of water, 190 ml of 0.2N
NaOH, 400 ml of water and 10.0 g of pancreatin. The
pH of the intestinal fluid was pH 7.5  0.1.
2.9. Preparation of digested sample solution
The lyophilized onion extracts were each submitted
to gastric or intestinal juice and a mutagen such as
IQ or MNNG. A 0.1 ml aliquot of each solution of
extract and mutagen was subjected to an equal volume
of gastric juice for 30 min. Another 0.1 ml aliquot of
each solution of extract and mutagen submitted to
an equal volume of intestinal juice for 2 h. This pro-
cess was performed in triplicate. Digested samples
were stored at 20
C until analysis in the mutagenic
activity.
662 M.-Y. Shon et al. / Food and Chemical Toxicology 42 (2004) 659–666

The antimutagenic activity of the lyophilized onion
extracts used in the experiments with the in vitro model
was tested by Ames test (Maron and Ames, 1983). The
antimutagenicities were used in the concentration of 3
mg per plate. As a mutagenic agent for TA98 and
TA100 was 0.2 mg for IQ and 0.1 mg for MNNG. For
each sample three agar plates were used and the number
of revertants per plate was counted with a colony
counter. Results of the Ames test were expressed as
mean number of revertants per triplicate plates, cor-
rected for spontaneous revertants.
2.10. Antimutagenic activity of digested onion extracts
The antimutagenic activity of the digested onion
extracts used in the experiments with the in vitro model
was tested by Ames test (Maron and Ames, 1983). This
procedure is same that used to measure antimutagenic
activity of the lyophilized onion extracts. All experi-
ments were performed in triplicate. Data presented in
the tales are means from three independent series.
In the DPPH test, the onion extracts were able to
reduce the stable radical DPPH to the yellow coloured
diphenylpicrylhydrazine. Methanol and hot water
extracts showed radical scavenging activity with 85% of
500 mg. The scavenging activity of radical on onion
extracts and BHT is presented in Fig. 1.
Hydrogen peroxide scavenging activity of the extract
is presented in Fig. 2. Scavenging activity of red onion
extract (10 mg/ml) and BHT(10 mg/l), l-ascorbic acid(10
mg/ml), reference compound, were 81% and 90%, 80%
of the initial concentration, respectively.
Fig. 3 shows the reducing power of the extracts using
the potassium ferricyanide reduction method. At 1 mg/
ml concentration, the extract obtained using red onion,
yellow onion and white onion showed absorbances of
0.17, 0.12, and 0.12, respectively. The l-ascorbic acid,
reference compound, exhibited the high reducing activ-
ity of 2.94 in 300 mg/ml.
Fig. 4 shows the antioxidant activity of the extracts as
measured by the bleaching of b-carotene. Extracts in red
onion, yellow onion and white onion as well as BHT,

3. Results

In total phenols and total flavonoids contents of

Table 1, red onion gave a high yields. The contents of
organic acids and free sugars are shown in Table 2. In
total free sugars content, all onion extracts gave about
600 mg/1 mg. In total organic acids content, yellow
onion gave a high yields.

Table 1
Contents of total phenolic compounds and flavonoids in ethyl acetate
extracts from three kinds of onion

(Dry base, m/mg)a

b
Onions Total phenols Total flavonoidsc
Fig. 1. Scavenging activities of ethyl acetate exrtracts of onion against
White 115 4 0.412 1,1-diphenyl-2-picrylhydrazl radical.
Red 133 7 0.511
Yellow 107 15 0.22
a
Each sample analyzed in triplicate
b
Total phenol contents based a standard curve generated by caffeic
acid.
c
Total flavonoids contents based a standard curve generated by
myricetin.

Table 2
Contents of total organic acids and free sugars in ethyl acetate extracts
from three kinds of onion

(Dry base, m/mg)

Onions Total organic acids Total free sugars

White 190.95a 629.14

Red 164.514 626.1212
Yellow 268.711 542.510
Fig. 2. Scavenging activities of ethyl acetate exrtacts of onion on
a
Data are presented as the meanS.D. of triplicate determinations. hydrogenperoxide.
 M.-Y. Shon et al. / Food and Chemical Toxicology 42 (2004) 659–666 663

l-ascorbic acid itself, were found to give the antioxidant
activity of 27%, 20%, 22%, 89%, and 37% at 1 mg/ml
concentration, respectively.
The onion extracts showed antimutagenicity against
IQ and MNNG in Salmonella typhimurium TA98 and
TA100 (Table 3). Welsh onion has already demon-
strated its effective antimutagenic activity (Huifeng et
al., 2001). Amounts of 3 mg per plate of onion extracts
were sufficient to inhibit the mutagenicity induced by
any of the IQ and MNNG concentrations used. These
results suggest that ethyl acetate extracts have anti-
mutagenic effects on both IQ and MNNG.
In Tables 4 and 5, digested onion extracts also showed
antimutagenicity. In all cases, concentration dependent
inhibition was observed. There were no significant dif-
ferences in antimutagenicity on the digested onion
extracts.

4. Discussion

Flavonoids and phenolic acids are the most pre-

Fig. 3. Reducing power of ethyl acetate extracts of onion.
Fig. 4. Antioxidant activities of ethyl acetate extracts of onion using
b-carotene linoleate model system.
dominant components of onion. These compounds have
antimutagenic potential activity. Onion extracts also
showed radical scavengers and antioxidant activities.
The onion extracts had phenolic hudroxyl groups in the
structure and antioxidants of phenolic groups have been
recognized to function as electron or hydrogen donors
(Shahidi and Wanasundara, 1992). The role of anti-
oxidants has attracted much interest with respect to
their protective effect against free radical damage that
may be the cause of many diseases including cancer
(Nakama et al., 1993). The antioxidative effect of onion
extract is mainly due to the phenolic components, such
as the flavonoids (Pietta et al., 1998). Some flavonoid
and non-flavonoid compounds have been reported to
also show alkylperoxyl radical scavenging activity thus
reducing radical-mediated pathogenesis, e.g. carcino-
genesis (Sawa et al., 1999).
Thus, the DPPH radical scavenging activity of onion
extracts may be mostly related to their phenolic
hydroxyl group. Also the concentration of hydrogen
peroxide in water may occur according to the phenolic
compounds and flavonoids. Since phenolic compounds
present in the extract are good electron donors, they may
accelerate the conversion of H2O2 to H2O (Ruch et al,
1984). As the good electron donors, they show the redu-
cing power. In Fig. 4, the antioxidant activity of car-
otenoids is based on the radical adducts of carotenoid

Table 3
Antimutagenic activity of ethyl acetate extracts from three kinds of on ion against IQ (0.02 mg, 0.2 mg/plate) and MNNG (0.1 mg, 0.01 mg/plate) on
Salmonella typhimurim TA98 and TA 100

Onions IQ MNNG
(3000 mg/plate)
Revertants/platea Percent inhibition (%) Revertants/plate Percent inhibition (%)

TA98 TA100 TA98 TA100 TA98 TA100 TA98 TA100

Control 152.712b 288.06 0 0 153.3 10 228.29 0 0

Red 59.35 154.52 61.12.9 46.43.3 57.0 13 127.15 62.8 2.5 44.3 3.6
White 47.37 145.010 69.00.8 49.75.6 57.2 6 133.02 62.7 2.3 41.7 0.4
Yellow 45.012 150.29 70.50.5 47.91.0 55.2 9 121.220 64.0 1.7 46.9 1.1
a
Triplicate plates were tested per dose per experiment.
b
Datas are presented as the mean S.D. of triplicate determinations.
664 M.-Y. Shon et al. / Food and Chemical Toxicology 42 (2004) 659–666

with free radicals from linoleic acid. The linoleic acid
free radical attacks the highly unsaturated b-carotene
models. The presence of carotenoids shows, not only a
decrease of the free radical concentration, but the
reduction of Fe3+ to Fe2+ by carotenoids. The pre-
sence of different antioxidants can hinder the extent of
b-carotene-bleaching by neutralizing the linoleate-free
radical and other free radicals formed in the system
(Jayaprakasha et al., 2001).
All types of onions in our study contained antimuta-
gens and the extracts showed similar levels of anti-
mutagenic activity. And antimutagenic effects of onion
extracts depended on the mutagen and dose levels.
Onions contain organosulfur compounds. Onion eth-
anol extracts also contain lipophilic antioxidant prop-
erties. It is known that onion oil contains dialkyl
disulfides and their oxides and thiols, which can trap
electrons from other systems. Thus it scavenges many
free radicals including hydroxyl radicals (Klanns-Dieter,
1983). Organosulfur compounds are also known to be
potent inducers of glutathione S-transferase (Hu et al.,
1997). Onion extracts maintained their antimutagenic
potential in vitro simulated digestion model. Thus,
antimutagenic compounds from onion extracts were not
inactivated by gastric acid and intestinal juices and were
readily available for absorption.

Table 4
Antimutagenic activity of ethyl acetate extracts from three kinds of onion against IQ (0.02 mg, 0.2 mg/plate) on Salmonella typhimurim TA98 and TA
100 under simulated gastric juice or intestinal juice

Onions Antimutagen Gastric juice Intestinal juice

(mg/plate)
Revertants/platea Percent inhibition (%) Revertants/plate Percent inhibition (%)

TA98 TA100 TA98 TA100 TA98 TA100 TA98 TA100

b
Red 0 228.35 307.33 0 0 217.07 602.31 0 0
1000 95.72 272.06 58.3 7 11.5 1.0 180.04 232.615 17.1 0.9 61.4 1.0
2000 59.713 223.017 74.1 1.6 27.4 0.7 101.312 207.63 53.3 1.7 65.5 1.3
3000 30.310 128.611 86.8 2.6 58.2 0.7 41.39 155.69 81.0 2.0 74.2 0.8
White 0 178.33 161.05 0 0 217.08 264.05 0 0
1000 100.07 130.02 43.9 1.0 20.0 0.4 115.55 117.04 46.8 2.0 55.7 0.9
2000 78.019 78.7 12 56.3 0.4 50.9 1.1 67.716 114.013 68.8 0.7 56.8 0.8
3000 32.32 75.0 4 81.9 0.4 53.4 1.7 57.320 95.3 7 73.6 0.8 63.9 1.6
Yellow 0 593.35 489.05 0 0 351.56 308.03 0 0
1000 128.77 246.13 78.3 1,5 46.7 1.8 73.24 124.07 79.2 1.1 59.7 1.8
2000 94.719 176.011 84.0 1.5 64.0 0.8 69.72 105.225 77.9 0.9 65.8 1.3
3000 80.020 160.216 86.5 0.9 67.3 1.6 56.211 92.1 11 84.0 1.6 70.0 0.8
a
Triplicate plates were tested per dose per experiment.
b
Datas are presented as the mean S.D. of triplicate determinations.

Table 5
Antimutagenic activity of ethyl acetate extracts from three kinds of onion against MNNG (0.1 mg, 0.01 mg/plate) on Salmonella typhimurim TA98
and TA 100 under simulated gastric juice or intestinal juice

Onions Antimutagen Gastric juice Intestinal juice

(mg/plate)
Revertants/platea Percent inhibition (%) Revertants/plate Percent inhibition (%)

TA98 TA100 TA98 TA100 TA98 TA100 TA98 TA100

b
Red 0 226.23 258.52 0 0 476.61 276.012 0 0
1000 167.75 144.36 25.9 1.2 44.2 1.0 321.53 137.53 32.5 0.4 50.2 1.0
2000 140.011 124.015 38.1 1.6 52.0 0.4 255.723 124.55 46.3 0.7 54.9 0.9
3000 124.58 108.53 45.0 0.4 58.0 0.3 179.510 103.16 62.3 1.3 62.6 0.7
White 0 217.33 260.39 0 0 229.03 241.52 0 0
1000 133.12 200.35 38.7 1.0 23.1 0.7 144.37 189.37 37.0 1.1 21.6 2.1
2000 115.010 180.214 47.0 0.4 30.8 0.4 118.59 134.615 48.3 0.8 44.3 0.9
3000 109.217 105.220 49.7 1.3 59.6 1.4 100.310 102.06 56.2 0.3 57.8 0.2
Yellow 0 148.09 197.011 0 0 155.53 252.09 0 0
1000 124.04 163.67 16.2 0.6 17.0 1.7 129.321 194.023 16.8 0.9 23.0 0.6
2000 106.71 133.04 27.9 2.5 32.5 1.1 118.76 171.14 23.7 1.4 32.1 0.8
3000 48.017 101.77 67.6 0.2 48.4 0.3 84.79 119.32 45.5 1.5 52.6 1.6
a
Triplicate plates were tested per dose per experiment.
b
Datas are presented as the mean S.D. of triplicate determinations.
 M.-Y. Shon et al. / Food and Chemical Toxicology 42 (2004) 659–666
IQ is a promutagen which can be converted to the
direct-acting mutagenic metabolite by cytochrome p450
isozymes, which are associated with IQ activation
(Beyeler et al., 1988). Flavonoids inhibited the direct
(nonactivated) mutagenicity of IQ in Salmonella strain
TA98. Flavonoids can also inhibit the mutagenicity
some of the cooking mutagens including IQ (Edenhar-
der et al., 1993). Flavonoids inhibit the mutation or
initiation caused by inhibition of activation of promu-
tagens and trap the electrophiles by chemical reaction or
conjugation; antioxidant activity or scavenging of reac-
tive oxygen species (De Flora, 1998).
Eventually human dietary studies will be required to
demonstrate that a coexposure to cooking mutagens
and flavonoids actually inhibits the activation and/or
DNA binding of the mutagens. The effect of dietary
flavonoids is that they can reduce the intestinal uptake
of IQ.
Antimutagenic activity against MNNG invested in
the present study was very low. This observation indi-
cates that these flavonoids are unable to interact and
neutralize electrophiles such as MNNG. Other papers
have also noted this failure and/or weak protective
effect against direct acting mutagens (Yamada and
Tomita 1994; Constable et al., 1996; Bu-Abbas et al.,
1994).
In conclusion, this study shows the antioxidant and
antimutagenic activity of onion extracts. But the anti-
mutagenic activity of onion extracts is measured in
relation to only two mutagens and despite the fact that
no biological materials are present in the model, it is
suggested that different inhibition mechanisms are
involved. Further research is needed to investigate the
efficacy of antimutagens in more detail, in particular
with respect to other types of mutagens.
In the present study, a weak relation exists between
the polyphenolic content of the onion extract and the
antimutagenic effect when using the indirect acting
mutagens IQ and the direct acting mutagen MNNG.
This study also suggested that onion extracts are useful
nutritional antioxidants and their supplementation nul-
lifies oxidative stress and onion extracts can serve as
such electron acceptors. When the onion extracts are
incubated, the protective effect could be ascribed to a
direct interaction of the activated mutagenic metabolite
that diffuses from the bottom layer. Therefore the
results of the present study show that the in vitro gas-
trointestinal model can be applied in mechanistic studies
on antimutagenicity in food and in research on the
development and efficacy of food products.
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