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Antiradical activity and polyphenol composition of local Brassicaceae edible

varieties

Article  in  Food Chemistry · December 2006

DOI: 10.1016/j.foodchem.2005.07.057

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Food
Chemistry
Food Chemistry xxx (2006) xxx–xxx
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2 Antiradical activity and polyphenol composition of local

3 Brassicaceae edible varieties

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4 Daniela Heimler a,*, Pamela Vignolini a, Maria Giulia Dini a,

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5 Franco Francesco Vincieri a,b, Annalisa Romani a,b
a
6 Department of Soil Science and Plant Nutrition, University of Florence, P.le delle Cascine 18, 50144 Florence, Italy
b
7 Department of Pharmaceutical Sciences, U. Schiff 6, 50019 Sesto F.no, Florence, Italy

Received 16 May 2005; accepted 27 July 2005

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10 Abstract

11
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The antiradical activity, polyphenols, flavonoids and total condensed tannins contents have been determined in the case of seven local
12 edible Brassicaceae, i.e. Italian kale, broccoli, Savoy and white cabbage, cauliflower, green cauliflower and Brussels sprouts. Rapid spec-
13 trophotometric methods were applied. The results achieved were compared with the quali–quantitative information obtained by HPLC/
14 DAD and HPLC/MS. The polyphenolic compounds detected were: kaempferol and quercetin glycosides and hydroxycinnamic esters.
15 The EC50 values ranged from 81.45 to 917.81 mg sample/mg DPPH and the total phenolic content from 4.30 to 13.80 gallic acid equiv-
16
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alents (mg gallic acid/g sample). The peculiar characteristics of these vegetables can be evaluated and can increase their value as func-
17 tional food.
18
2005 Published by Elsevier Ltd.

19 Keywords: Brassicaceae; Polyphenols; Flavonoids; Antiradical activity; Functional food

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20
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21 1. Introduction Many epidemiological studies have correlated the intake 37

of a diet rich in fruits and vegetables with a reduced risk of 38
22 The Brassicaceae family includes more than 350 genera incidence of chronic diseases, such as cancer and cardiovas- 39
23 and 3500 species, for the majority cool season annuals, cular disease. In particular, several epidemiological studies 40
24 characterised by short cycle and wide adaptability; for this report an inverse correlation between consumption of Brass- 41
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25 reason they are suited for cultivation in different seasons icaceae and risk of cancer (Peto, Doll, Buckley, & Sporn, 42
26 and in a variety of environments. 1981; Stoewsand, Anderson, & Munson, 1998) probably 43
27 As regards to the nutritional profile, the Brassicaceae due to the anticancer action of metabolites of glucosinolates, 44
28 have a low caloric value (24–34 kcal/100 g) depending on as demonstrated by some ‘‘in vitro’’ studies (Verhoeven, Ver- 45
29 the low content of protein (1.44–2.82/100 g) and fat hagen, Goldbohm, Van den Brandt, & Van Poppel, 1997). 46
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30 (0.12–0.37/100 g) and an average content of fibre of 2.5/ Nevertheless, we cannot exclude that the protective effect 47
31 100 g. On the contrary, the contents of minerals, vitamins against chronic diseases could also depend on the antioxidant 48
32 and others phytochemicals such as polyphenols and gluco- activity of other compounds present in those vegetables, such 49
33 sinolates, sulphur containing compounds, are notable. as vitamin C, polyphenols, vitamin E and carotenoids (Byers 50
34 These vegetables are rich in potassium, calcium, magne- & Perry, 1992; Evangelou et al., 1997). A more accurate char- 51
35 sium and phosphorus, vitamins C, E, K and carotenoids acterization of antioxidant levels of some varieties of 52
36 (b-carotene, lutein and zeaxanthin). Brassicaceae seems, therefore, opportune, on the basis of 53
their cancer preventive properties (Beecher, 1994). 54
*
Corresponding author. Considering the chemical diversity of the antioxidant 55
E-mail address: daniela.heimler@unifi.it (D. Heimler). compounds present in foods and the interaction occurring 56

0308-8146/$ - see front matter
2005 Published by Elsevier Ltd.

doi:10.1016/j.foodchem.2005.07.057
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57 among those different molecules, the evaluation of the total at least; all data are mean values (standard deviation within 111
58 antioxidant capacity of foods seems to be a more useful brackets).The edible part of each vegetable (the florets in 112
59 marker than the evaluation of single compound. However, the case of broccoli) was frozen in liquid nitrogen and 113
60 no single method to test the total antioxidant capacity of stored at
80 C before proceeding with the analysis. Fro- 114
61 foods (TRAP, ORAC, etc.) fully consider, at the same zen tissues were then ground in a mortar with a pestle un- 115
62 time, the activity of all the antioxidant compounds. der liquid nitrogen. A quantity of 1.5 g of tissue was 116
63 A possible approach could be to consider the antiradical extracted in 20 mL of 70% ethanol (pH 3.2, by formic acid) 117
64 activity together with the polyphenols content. overnight. This solution was used for the determination of 118
65 Plant polyphenols are known to have multifunctional antioxidant activity, total phenolic, and flavonoid contents. 119
66 properties such as reducing agents, hydrogen donating For condensed tannins content, the solution was evapo- 120

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67 antioxidants and singlet oxygen quenchers and flavonoids rated to dryness under reduced pressure at room tempera- 121
68 and their derivatives are the largest and most important ture by a Rotavapor 144 R, Büchi, (Switzerland) and 122

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69 group of polyphenols. The most important property is their finally rinsed with a CH3CN/CH3OH/H2O (pH 2, by 123
70 capacity to act as antioxidants protecting the body against HCOOH) 60:20:20 mixture to a final volume of 2 mL. 124
71 reactive oxygen species and may have an additive effect to For HPLC analysis, the solution was treated as before with 125
72 the endogenous scavenging compounds (Rice-Evans, Mill- the only exception of the addition, before the evaporation, 126
73 er, & Paganga, 1997). The antioxidative effect is performed of 40 lL gallic acid (5.88 mM) as internal standard; the 127

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74 through different mechanisms the most general and impor- concentrated solution was used after an extraction step 128
75 tant of which is direct radical scavenging which depends on with n-hexane. The extraction yield (95%) was controlled 129
76 the chemical structure of the flavonoids involved (Nijveldt by the addition of gallic acid which is not naturally present 130
77 et al., 2001). It is generally accepted that this action leads to in our samples and exhibits a retention time which falls in 131
78 afford protection against numerous chronic diseases, an empty zone of the chromatogram. 132
79 including cancer, cardio- and cerebrovascular, ocular and Authentic standards of rutin, clorogenic acid, gallic 133
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80 neurological diseases (Block, Patterson, & Subhar, 1992; acid, cathechin, Folin–Ciocalteau reagent, and 1,1-diphe- 134
81 Youdim & Joseph, 2001). nyl-2-picrylhydrazil radical (DPPH) were purchased from 135
82 Brassicaceae are known to contain flavonoids, and espe- Sigma–Aldrich. 136
83 cially flavonols (Nielsen, Norbaek, & Olsen, 1998; Price, All solvents were of HPLC grade purity (BDH Labora- 137
84 Casuscelli, Colquhoun, & Rhodes, 1998; Vallejo, Tomas- tory Supplies, United Kingdom). 138
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85 Barberan, & Garcia-Viguera, 2002) and their antioxidant

86 activity has been assessed in some cases (Chu, Chang, & 2.2. Antioxidant activity 139
87 Hsu, 2000; Kaur & Kapoor, 2002); and correlated to can-
88 cer preventive properties (Beecher, 1994). Free radical scavenging activity was evaluated with the 140
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89 The aim of this paper is the study of the antiradical DPPH (1,1-diphenyl-2-picrylhydrazil radical) assay. The 141
90 activity in correlation to the polyphenolic content of edible antiradical capacity of the sample extracts was estimated 142
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91 parts of some varieties of Brassica oleracea, among which according to the procedure reported by Brand-Williams 143
92 also local varieties were considered, which are largely con- and Cuvelier (1995) and slightly modified. 144
93 sumed in Italy, i.e. white cabbage, broccoli, Italian kale Two milliliter of the sample solution, suitably diluted 145
94 (black cabbage), Savoy cabbage, cauliflower, green cauli- with ethanol, was added to 2 mL of an ethanol solution 146
95 flower, and Brussels sprouts. The comparison of different of DPPH (0.0025 g/100 mL) and the mixture was allowed 147
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96 varieties used in one country is interesting from a nutri- to stand. After 20 min, the absorption was measured at 517 148
97 tional point of view. nm (LAMBDA 25, Perkin–Elmer spectrophotometer) ver- 149
sus ethanol, as a blank. Each day, a calibration curve of 150
98 2. Experimental DPPH was carried out. The antioxidant activity is ex- 151
pressed as EC50, the antioxidant dose required to cause a 152
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99 2.1. Sample preparation 50% inhibition (Sanchez-Moreno, Larrauri, & Saura-Cali- 153
xto, 1998). EC50 was calculated plotting the ratio 154
100 The following vegetables were purchased from local (DPPHrem): [DPPH concentration at t = 20 0 ]/[DPPH 155
101 markets on February 2004, that is the period in which concentration at t = 0] · 100 against the concentration of 156
102 in Italy cabbages are the most representative vegetables: the antioxidant. EC50 is expressed as mg antioxidant/mg 157
103 white cabbage (B. oleracea L. var. capitata L.), broccoli DPPH. 158
104 (B. oleracea L. conv. botrytis L. var. italica Plenk), Italian
105 Kale (B. oleracea L. var. acephala D C.), Savoy cabbage (B. 2.3. Total phenolic content 159
106 oleracea L. var. sabauda L.), green cauliflower (B. oleracea
107 L. conv. botrytis L. var. botrytis cv Verde di Macerata), The total phenolic content was determined using the 160
108 cauliflower (B. oleracea L. conv. botrytis L. var. botrytis Folin–Ciocalteu method, described by Singleton, Orthofer, 161
109 cv Snow ball), and Brussels sprouts (B. oleracea L. var. and Lamuela-Raventos (1999) and slightly modified 162
110 gemmifera Zencher). Each experiment was run three times according to Dewanto, Wu, Adom, and Liu (2002). To 163
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164 125 lL of the suitably diluted sample extract, 0.5 mL of Table 1

165 deionised water and 125 lL of the Folin–Ciocalteu reagent The linear solvent gradient system used in HPLC–DAD and HPLC–MS
analysis of polyphenols
166 were added. The mixture was allowed to stand for 6 min
167 and then 1.25 mL of a 7% aqueous Na2CO3 solution were Time (min) H2O/H+ (%) CH3CN (%)
168 added. The final volume was adjusted to 3 mL. The mix- 0 98 2
169 ture was allowed to stand for 90 min and the absorption 16 87 13
23 87 13
170 was measured at 760 nm against water as a blank. The 33 75 25
171 amount of total phenolics is expressed as gallic acid equiv- 42 0 100
172 alents (GAE, mg gallic acid/g sample) through the calibra- 50 0 100
173 tion curve of gallic acid. The calibration curve ranged 20–

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174 500 lg/mL (R2 = 0.9969).
2.7. HPLC/MS analysis 215

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175 2.4. Total flavonoid content
Analyses were performed using a HP 1100L liquid chro- 216
176 The total flavonoid content was determined using a matograph linked to a HP 1100 MSD mass spectrometer 217
177 colorimetric method described by Dewanto et al. (2002) with an API/electrospray interface (Agilent Technologies, 218
178 and slightly modified in our laboratory. To 0.25 mL of Palo Alto, CA, USA). The mass spectrometer operating 219
conditions were: gas temperature, 350 C; nitrogen flow 220

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179 the suitably diluted sample, 75 lL of a 5% NaNO2 solu-
180 tion, 0.150 mL of a freshly prepared 10% AlCl3 solution, rate, 10.0 L/min, nebulizer pressure, 40 psi; quadrupole 221
181 and 0.5 mL of 1 M NaOH solution were added. The final temperature, 40 C; and capillary voltage, 3500 V. The 222
182 volume was adjusted to 2.5 mL with deionised water. The mass spectrometer was operated in positive and negative 223
183 mixture was allowed to stand for 5 min and the absorp- mode at 80–120 eV. 224
184 tion was measured at 510 nm against the same mixture, The identity of polyphenols was ascertained using data 225
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185 without the sample, as a blank. The amount of total from HPLC/DAD and HPLC/MS analyses by comparison 226
186 flavonoids is expressed as (+)catechin equivalents (CE, and combination of their retention times, UV/Vis, and 227
187 mg (+)catechin/g sample) through the calibration curve mass spectra with those of authentic standards. Quantifica- 228
188 of (+)catechin. The calibration curve ranged 10–500 lg/ tion of individual polyphenolic compounds was directly 229
189 mL (R2 = 0.9946). performed by HPLC/DAD using a five-point regression 230
CT

curve (R2 P 0.998) in the range 0–5 lg on the basis of 231

190 2.5. Total condensed tannins authentic standards. 232

191 The analysis of condensed tannins (procyanidins) was 3. Results and discussion 233
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192 carried out according to the method of Broadhurst and

193 Jones (1978) and slightly modified in our laboratory. To Table 2 lists the total phenolic content (expressed as gal- 234
lic acid equivalents), the flavonoids content (expressed as 235
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194 50 lL of the suitably diluted sample, 3 mL of a 4% metha-

195 nol vanillin solution and 1.5 mL of concentrated hydro- catechin equivalents), the tannins content (expressed as cat- 236
196 chloric acid were added. The mixture was allowed to echin equivalents) and the antiradical activity (expressed as 237
197 stand for 15 min and the absorption was measured at EC50). Broccoli and Italian kale exhibit the highest content 238
198 500 nm against methanol as a blank. The amount of total of both total phenolics and flavonoids. The amounts of to- 239
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199 condensed tannins is expressed as (+)catechin equivalents tal phenolics are very similar to those found with the same 240
200 (CT, mg (+)catechin/g sample). The calibration curve ran- method by Kaur and Kapoor (2002) in the case of broccoli 241
201 ged 50–600 lg/mL (R2 = 0.9978). No absorbance was ob- and Brussels sprouts and by Chun, Smith, Sakawaga, and 242
202 tained without vanillin addition. Lee (2004) for Savoy cabbage. The highest tannins content 243
is shown by cauliflower. 244
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203 2.6. HPLC/DAD analysis There is a good correlation (R2 = 0.974) between the to- 245
tal phenolics and the flavonoids content. 246
204 Analysis of flavonols and phenolic acids were carried As regards the EC50 values broccoli and Italian kale ex- 247
205 out using a HP 1100L liquid chromatograph equipped hibit the lowest value (from 4 to 23 times lower than what 248
206 with a DAD detector and managed by a HP 9000 work- found for all other vegetables). 249
207 station (Agilent Technologies, Palo Alto, CA, USA). As regards the correlation between EC50 values and 250
208 Flavonols and phenolic acids were separated by using a phenolics and flavonoids content, the contribution of vita- 251
209 50 · 2.2 mm i.d. 3 lm Luna C18 column (Waters) operat- min C should be taken into account. In some B. oleracea 252
210 ing at 26 C, according to the linear solvent gradient sys- subspecies (broccoli, cauliflower, cabbage) the content of 253
211 tem of Table 1 during a 50 min period. UV/Vis spectra vitamin C (an antioxidant molecule which was not evalu- 254
212 were recorded in the 190–600 nm range and the chro- ated separately in this paper) changed from 27.32 to 255
213 matograms were acquired at 260, 280, 305, 330 and 74.71 mg/100 g fresh weight (Kurilich et al., 1999). Kuri- 256
214 350 nm. lich, Jeffery, Juvik, Wallig, and Klein (2002) in a paper 257
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Table 2
Total phenolics, flavonoids, and tannins contents; EC50 values
Sample Total phenolics gallic acid/sample Flavonoids (+)-catechin/sample Tannins (+)-catechin/sample EC50 sample/DPPH
(mg/g, dry weight) (mg/g, dry weight) (mg/g, dry weight) (mg/mg)
White cabbage 5.31 (0.906) 1.98 (0.067) 0.50(0.005) 370.67 (13.576)
Broccoli 12.85 (0.195) 6.71 (0.467) 0.41(0.033) 81.45 (1.919)
Italian kale 13.80 (0.165) 5.09 (0.540) 0.39(0.002) 92.91 (2.651)
Savoy cabbage 4.30 (0.242) 1.35 (0.003) 0.38(0.006) 379.96 (27.442)
Green cauliflower 6.52 (0.772) 2.52 (0.062) 0.48(0.015) 354.47 (20.796)
Cauliflower 5.83 (0.133) 1.69 (0.522) 0.56(0.073) 917.81 (24.114)
Brussels sprouts 8.10 (0.219) 3.07 (0.262) 0.50(0.005) 339.57 (22.848)

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Standard deviation within brackets.

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258 concerning different broccoli genotypes did not find any tion with TEAC (trolox equivalent antioxidant capacity, 271
259 correlation between the antioxidant activity and the flavo- determined with the ABTS radical). 272
260 noids or vitamin C contents while a correlation was found The DPPH method, on a molar base, does not give the 273
261 between the antioxidant activity and the lutein and zeaxan- same results depending on the standard used (Butkovic, 274
262 thin contents, which are not extracted under our experi- Klasinc, & Bors, 2004) owing to different kinetic in the H 275

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263 mental conditions. On the other side Chun et al. (2004) atom transfer (Goupy, Dufour, Loonis, & Dangles, 276
264 found good correlation (R2 P 0.95) between antioxidant 2003); in fact with our method the EC50 value (expressed 277
265 activity and total flavonoid and total phenolic contents as lmoles of standard/mg DPPH) is 1.18 for ascorbic acid, 278
266 for raw (red cabbage, green cabbage, Napa cabbage and 2.80 for kaempferol, 1.15 for quercetin and 2.03 for querci- 279
267 Savoy cabbage) and processed (sauerkraut products) cab- trin. Comparing the EC50 values with the antioxidant activ- 280
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268 bages using an ABTS radical solution to asses the antioxi- ity (Proteggente et al., 2002) expressed as trolox equivalent 281
269 dant activity. Proteggente et al. (2002) found that total antioxidant capacity (TEAC), ferric reducing ability of 282
270 phenolic and vitamin C contents showed a good correla- plasma (FRAP), (ORAC) oxygen radical absorbance 283
CT

350 nm 1
mAU 4
800
a

600
2 3
400
k 6 kk 8
*
E

200 5 k Q 7
0
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10 20 30 40 50 min

mAU 350 nm 8
1400
1200 11 b
1000
800 7 9 10
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600 12
400 k
200 Qk k 6
0
10 20 30 40 50 min

mAU 280 nm
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200
175
c
150 IS
125 *
100
75
50
25
0
10 20 30 40 50 min

Fig. 1. Chromatographic profiles acquired by HPLC/DAD (350 nm) of the hydroalcholic (ethanol:water 70:30, pH 2) extracts of: (a) Italian kale; (b)
Broccoli florets and (c) Brussels sprouts. Polyphenolic compounds: 1. Kaempferol-3-[2-sinapoylglucopiranosyl (1,2) glucopiranoside]-7-[glucopiranosyl
(1,4) glucopiranoside]; 2. Kaempferol tetraglucoside; 3. Kaempferol sinapoyl tetraglucoside; 4. Kaempferol-3-[-2-feruloylglucopiranosyl (1,2)
glucopiranoside]-7-[glucopiranosyl (1, 4) glucopiranoside]; 5. Kaempferol cumaroyl tetraglucoside; 6. Kaempferol diglucoside; 7. 1,2-disinapoylgenti-
obiose; 8. 1 sinapoyl-2-feruloylgentiobiose; 9. 1,2-diferuloylgentiobiose; 10. 1,2,2 0 -trisinapoyl gentiobiose; 11. 1,2-disinapoyl-2-feruloylgentiobiose; 12. 1-
sinapoyl-2,2 0 -diferuloylgentiobiose. *condensed tannins (ipothesis) identified at 280 nm; IS, internal standard (gallic acid); K, kaempferol derivatives, Q,
quercetin derivatives; C, caffeic acid derivative (chlorogenic acid).
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284 capacity, the same hierarchy is found for broccoli, cabbage Table 3
285 (a mixture of Savoy and white) and cauliflower indicating Flavonoids and phenolic acids content (mg/g, dry weight) as obtained
from HPLC measurements
286 that the DPPH method reflects the results obtained with
287 different experimental approaches. Sample Flavonoids Phenolic acids Total polyphenols
288 If we consider all vegetables with the exception of cauli- White cabbage 2.70 0.07 2.77
289 flower, a correlation is found between both antioxidant Broccoli 3.04 8.69 11.73
Italian kale 11.27 n.d. 11.27
290 activity and total phenolic content (R2 = 0.9388) and anti- Savoy cabbage 1.02 0.23 1.25
291 oxidant activity and flavonoids content (R2 = 0.9131). Green cauliflower 2.10 0.07 2.17
292 Cauliflower EC50 high value (917.81 mg/mg DPPH) indi- Cauliflower 0.29 0.09 0.38
293 cates that the antiradical activity of this vegetable is partic- Brussels sprouts 1.12 0.35 1.47

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294 ularly low, and the difference between cauliflower and
295 green cauliflower, notwithstanding the belongings to the
parameter in assessing the antiradical activity of these vege- 341

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296 same subspecies, is probably due to the edible part which
tables can be drawn from its large range of variation (about 342
297 does not include the green leaves.
100 times) with respect to other tests such as total polyphe- 343
298 As regards polyphenol characterization, as an example,
nols and flavonoids (ranges of variation about 3–5 times). 344
299 Fig. 1 reports the chromatographic profiles recorded at
300 350 nm of the extracts of: (a) Italian kale, (b) broccoli
Acknowledgements 345

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301 and (c) Brussels sprouts. In these extracts, both flavonoids
302 and hydroxycinnamic derivatives were identified by their
The authors express their sincere gratitude to the Cassa 346
303 chromatographic behaviour, UV spectra, MS spectra,
di Risparmio di Firenze that contributed to the acquisition 347
304 retention times in comparison with authentic standards.
of a part of the instrumentation used for this work. 348
305 Among flavonols, we identified kaempferol-3-[2-sinapoylg-
306 lucopyranosyl (1,2) glucopyranoside]-7-[glucopyranosyl
References 349
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307 (1,4) glucopyranoside], kaempferol-3-[-2-feruloylglucopyr-
308 anosyl (1,2) glucopyranoside]-7-[glucopyranosyl (1,4) Beecher, C. W. W. (1994). Cancer preventive properties of varieties of 350
309 glucopyranoside] (Nielsen et al., 1998), kaempferol tetrag- Brassica oleracea: a review. American Journal of Clinical Nutrition, 59, 351
310 lucoside, kaempferol sinapoyl tetraglucoside, kaempferol 1166–1170. 352
311 cumaroyl tetraglucoside, kaempferol diglucoside (Romani Block, G., Patterson, B., & Subhar, A. (1992). Fruit, vegetables and cancer 353
CT

312 et al., 2003). The predominant hydroxycinnamic acids were prevention: a review of the epidemiological evidence. Nutrition and 354
Cancer, 18, 1–29. 355
313 identified as 1,2-disinapoylgentiobiose, 1-sinapoyl-2-feru- Brand-Williams, W., & Cuvelier, M. E. (1995). Use of a free radical 356
314 loylgentiobiose, 1,2-diferuloylgentiobiose, 1,2,2 0 -trisina- method to evaluate the antioxidant activity. Lebensmittel-Wissenshaft 357
315 poyl gentiobiose, 1,2-disinapoyl-2-feruloylgentiobiose, und Technologie, 28, 25–30. 358
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316 1-sinapoyl-2,2 0 -diferuloylgentiobiose; the pattern found Broadhurst, R. B., & Jones, W. T. (1978). Analysis of condensed tannins 359
317 for hydroxycinnamic derivatives in broccoli florets was using acified vanillin. Journal of the Science of Food and Agriculture, 360
29, 788–794. 361
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318 similar to that described by Vallejo, Tomas-Barberan, Butkovic, V., Klasinc, L., & Bors, W. (2004). Kinetic study of flavonoid 362
319 and Garcia-Viguera (2003). reactions with stable radicals. Journal of Agricultural and Food 363
320 Italian kale (black cabbage) polyphenols composition Chemistry, 52, 2816–2820. 364
321 has already been studied in relation to the environment Byers, T., & Perry, G. (1992). Dietary carotenes, vitamin C and vitamin E 365
322 (Vallejo et al., 2003). as protective antioxidants in human cancers. Annual Review of 366
Nutrition, 12, 139–159. 367
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323 Brussels sprouts as well as all other samples, with the Chu, Y. H., Chang, C. L., & Hsu, H. F. (2000). Flavonoid content of 368
324 exception of Italian kale and broccoli, exhibit a very poor several vegetables and their antioxidant activity. Journal of the Science 369
325 profile and the attribution of peaks, even if HPLC/DAD of Food and Agriculture, 80, 561–566. 370
326 and HPLC/MS are employed, is difficult and, overall, the Chun, O. K., Smith, N., Sakawaga, A., & Lee, C. Y. (2004). Antioxidant 371
327 quantitative data are affected by the shape of the properties of raw and processed cabbages. International Journal of 372
Food Science and Technology, 55, 191–199. 373
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328 chromatogram. Dewanto, V., Wu, X., Adom, K. K., & Liu, R. H. (2002). Thermal 374
329 Table 3 lists the quantitative data from HPLC/DAD processing enhances the nutritional value of tomatoes by increasing 375
330 measurements. There is a quite good correlation total antioxidant activity. Journal of Agricultural and Food Chemistry, 376
331 (R2 = 0.8801) between total polyphenols from HPLC data 50, 3010–3014. 377
332 and total flavonoids from AlCl3 test (see Table 2). It should Evangelou, A., Kalpouzos, G., Karkabounas, S., Liasko, R., Nonni, A., 378
Stefanou, D., et al. (1997). Dose-related preventive and therapeutic 379
333 be noted that, owing to the shape of chromatograms, all effect of antioxidants-anticarcinogens on experimentally induced 380
334 flavonoids have been expressed as rutin content through malignant tumours in Wirstar rats. Cancer Letters, 115, 105–111. 381
335 a calibration curve; this occurrence may affect the results, Goupy, P., Dufour, C., Loonis, M., & Dangles, O. (2003). Quantitative 382
336 especially when small amounts are considered. kinetic analysis of hydrogen transfer reactions from dietary polyphe- 383
337 Our results show how different Brassicaeae can be evalu- nols to the DPPH radical. Journal of Agricultural and Food Chemistry, 384
51, 615–622. 385
338 ated for their polyphenols content and their antiradical Kaur, C., & Kapoor, H. C. (2002). Anti-oxidant activity and total 386
339 activity giving more information on the peculiar functional phenolic content of some Asian vegetables. International Journal of 387
340 characteristics of each food. The importance of the EC50 Food Science and Technology, 37, 153–161. 388
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389 Kurilich, A. C., Jeffery, E. H., Juvik, J. A., Wallig, M. A., & Klein, B. P. and at different elevations. Italian Journal of Food Science, 15, 417
390 (2002). Antioxidant capacity of different broccoli (Brassica oleracea) 197–205. 418
391 genotypes using the oxygen radical absorbance capacity (ORAC) Sanchez-Moreno, C., Larrauri, J. A., & Saura-Calixto, F. (1998). A 419
392 assay. Journal of Agricultural and Food Chemistry, 50, 5053–5057. procedure to measure the antiradical efficiency of polyphenols. Journal 420
393 Kurilich, A. C., Tsau, G. J., Brown, A., Howard, L., Klein, B. P., Jeffery, of the Science of Food and Agriculture, 76, 270–276. 421
394 E. H., et al. (1999). Carotene, tocopherol and ascorbate contents in Singleton, V. L., Orthofer, R., & Lamuela-Raventos, R. M. (1999). 422
395 subspecies of Brassica oleracea. Journal of Agricultural and Food Analysis of total phenols and other oxidation substrates and antiox- 423
396 Chemistry, 47, 1576–1581. idants by means of the Folin–Ciocalteu reagent. Methods in Enzymol- 424
397 Nielsen, J. K., Norbaek, R., & Olsen, C. E. (1998). Kaempferol ogy, 299, 152–178. 425
398 tetraglucosides from cabbage leaves. Phytochemistry, 49, 2171–2176. Stoewsand, G., Anderson, J., & Munson, L. (1998). Protective effects of 426
399 Nijveldt, R. J., Van Nood, E., Van Hoorn, D. E. C., Boelens, P. G., Van dietary Brussels sprouts against mammary carcinogenesis in Sprague- 427
400 Norren, K., & Van Leeuwen, P. A. M. (2001). Flavonoids: a review of Dawley rats. Cancer Letters, 39, 199–207. 428

F
401 probable mechanism of action and potential applications. American Vallejo, F., Tomas-Barberan, F. A., & Garcia-Viguera, C. (2003). 429
402 Journal of Clinical Nutrition, 74, 418–425. Phenolic compound contents in edible parts of broccoli inflorescences 430
403 Peto, R., Doll, R., Buckley, J., & Sporn, M. (1981). Can dietary b-carotene after domestic cooking. Journal of the Science of Food and Agriculture, 431

OO
404 materially reduce human cancer rates? Nature, 290, 201–208. 83, 1511–1516. 432
405 Price, K. R., Casuscelli, F., Colquhoun, I. J., & Rhodes, M. J. C. (1998). Vallejo, F., Tomas-Barberan, F. A., & Garcia-Viguera, C. (2002). 433
406 Composition and content of flavonols glycosides in broccoli florets. Potential bioactive compounds in health promotion from broccoli 434
407 Journal of the Science of Food and Agriculture, 77, 468–472. cultivars grown in Spain. Journal of the Science of Food and 435
408 Proteggente, A. R., Pannala, A. S., Pagana, G., Van Buren, L., Wagner, E., Agriculture, 82, 1293–1297. 436
409 Wiseman, S., Van De Put, F., & Dacombe, C. (2002). The antioxidant Verhoeven, D. T. H., Verhagen, H., Goldbohm, R. A., Van den Brandt, P. 437

PR
410 activity of regularly consumed fruit and vegetables reflects their phenolic A., & Van Poppel, G. A. (1997). Review of mechanism underlying 438
411 and vitamin C composition. Free Radical Research, 36, 217–233. anticarcinogenicity by Brassica vegetables. Chemico-Biological Inter- 439
412 Rice-Evans, C. A., Miller, N. J., & Paganga, G. (1997). Antioxidant actions, 103, 79–129. 440
413 properties of phenolic compounds. Trends in Plant Science, 2, 152–159. Youdim, K. A., & Joseph, J. A. A. (2001). A possible emerging role of 441
414 Romani, A., Pinelli, P., Galardi, C., Corti, G., Agnelli, A., Vincieri, F. F., phytochemicals in improving age-related neurobiological dysfunctions: 442
415 et al. (2003). Flavonoids in leaves of black cabbage (Brassica oleracea a multiplicity of effects. Free Radical Biology and Medicine, 30, 443
416 var. acephala DC. subvar. viridis cv. serotina) grown on different soils 583–594. 444
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