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LWT - Food Science and Technology 41 (2008) 2118e2125 www.elsevier.com/locate/lwt

Antioxidant properties of durian fruit as inuenced by ripening

Patricia Arancibia-Avila a, Fernando Toledo a, Yong-Seo Park b, Soon-Teck Jung c, Seong-Gook Kang c, Buk Gu Heo d, Sang-Hyun Lee e, Mietek Sajewicz f, Teresa Kowalska f, Shela Gorinstein g,*,1
a

Department of Basic Sciences, Universidad del Bio-Bio, Campus Fernando May, P.O. BOX 447, Chillan, Chile b Department of Horticultural Science, Mokpo National University, Muan, Jeonnam, South Korea c Department of Food Engineering, Mokpo National University, Muan, Jeonnam, South Korea d Naju Foundation of Natural Dyeing Cultural Institute, Naju, South Korea e Pear Experiment Station, RDA, Naju, Jeonnam, South Korea f Institute of Chemistry, Silesian University, Katowice, Poland g Department of Medicinal Chemistry and Natural Products, School of Pharmacy, The Hebrew University e Hadassah Medical School, Jerusalem, Israel Received 22 April 2007; received in revised form 29 July 2007; accepted 4 December 2007

Abstract The antioxidant properties of durian (Durio zibethinus Murr., cv. Mon Thong) at different stages of ripening were investigated using uorometry, UV spectroscopy, and HPLC/DAD analyses. Total polyphenols, avonoids, anthocyanins and avanols in ripe durian were signicantly higher ( p < 0.05) than in mature and overripe fruits. Free polyphenols and avonoids were at lower levels than hydrolyzed ones. Caffeic acid and quercetin were the dominant antioxidant substances in ripe durian. In these fruits, methanol extracts contained a relatively high capacity of 74.9 7.1% inhibition using b-caroteneelinoleic acid assay. Ferric-reducing/antioxidant power (FRAP) and cupric-reducing antioxidant capacity (CUPRAC) assays supported this nding. The correlation coefcients between polyphenols and antioxidant capacities of durian samples with all applied assays were about 0.98. In conclusion, the bioactivity of ripe durian was high and the total polyphenols were the main contributors to the overall antioxidant capacity. 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.
Keywords: Ripe, overripe and mature durian; Bioactive compounds; Antioxidant capacity

1. Introduction The health protective effect of natural products such as fruits and vegetables is mostly related to their antioxidants, phenolic compounds, and to a lesser extent, dietary ber (Chun et al., 2005; Dauchet, Amouyel, Hercberg, & Dallongeville, 2006; Erkkilae, Herrington, Mozaffarian, & Lichtenstein, 2005; Jung, Su, Keller, Mehta, & Kinghorn, 2006; Koebnick et al., 2005; Lairon et al., 2005; Mahattanatawee et al., 2006). Among
* Corresponding author. Tel.: 972 2 6758690; fax: 972 2 6757076. E-mail address: [email protected] (S. Gorinstein). 1 S.G. is afliated with the David R. Bloom Center for Pharmacy.

these fruits is the lesser known durian [D. zibethinus Murr. cv. Mon Tong] (Ketsa & Daengkanit, 1998). Ketsa and Daengkanit (1998) studied postharvest changes in ethylene production, respiration, solids, total sugars, starch, rmness, pectic substances and activities of polygalacturonase (PG), pectinesterase (PE) of durian, but did not evaluate antioxidant properties. Durian is consumed at different stages of ripening, and the differences in nutritional quality between ripening stages are practically unknown. Some authors have shown that there are signicant differences in the content of bioactive compounds and in the antioxidant capacity of other tropical fruits at various stages of their ripening (Park et al., 2006; Zhang, Koo, & Eun, 2006). As far as we know, no

0023-6438/$34.00 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.lwt.2007.12.001

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studies of the antioxidant capacity of durian have been conducted, and there are no published articles describing these properties of durian at different stages of its ripening. Therefore, our objective was to study one of the most popular cultivar of durian Mon Thong in vitro at different stages of its ripening. In order to receive a reliable picture of the differences between the mature, ripe and overripe samples of durian, the major antioxidant compounds (polyphenols, avonoids, avanols and anthocyanins) were determined (Cheng & Breen, 1991; Singleton, Orthofer, & Lamuela-Raventos, 1999; Vinson, Su, Zubic, & Bose, 2001). It was shown that the measures of the antioxidant capacity in natural products by only one assay are often not reliable (Ou, Huang, Hampsch-Woodill, Flanagan, & Deemer, 2002). Therefore, in this investigation we used three complementary assays. 1. Antioxidant test using b-caroteneelinoleate model system [b-carotene] (Ferreira, Proenca, Serralheiro, & Araujo, 2006) 2. Ferric-reducing/antioxidant power [FRAP] (Szeto, Tomlinson, & Benzie, 2002) 3. Cupric-reducing antioxidant capacity [CUPRAC] (Apak, Guclu, Ozyurek, & Karademir, 2004) 2. Materials and methods 2.1. Samples In this investigation, the Mon Thong cultivar at different stages of ripening was studied. Harvesting and determination of maturity was carried out by Thai skilled workers, combining the following techniques: day count, character of fruit spines, tapping the fruit, color and shape of fruit (Yaacob & Subhadrabandhu, 1995). The mature durian fruits were harvested carefully with peduncle intact. The samples were left for 1 day at room temperature and cut open to get mature durian esh with rm texture and no smell. Some of the fruits were left for another 4 days at room temperature to ripen until their esh became soft and they developed a typical durian aroma. Overripe samples having a strong smell were obtained when fruits were left for another 3 days. The edible parts, botanically called aril, of the Mon Thong at different stages of ripening were prepared without using steel knives. The fruits were cleaned with tap water, dried, weighed, chopped and homogenized under liquid nitrogen in a high-speed blender (Hamilton Beach Silex professional model) for 1 min. A weighed portion (50e100 g) was lyophilized for 48 h (Virtis model 10-324), and the dry weight was determined. The samples were ground to pass through a 0.5-mm sieve and stored at 20  C until analyzed. 2.2. Extract procedures Durian lyophilized samples (0.2 g) were placed in a small vial and 3 ml of a binary mixture composed of ethanol and 0.2 M HCl (1:1, v/v) was added. The samples were kept in

RK 255 H Sonorex Super sonication bath manufactured by the rm Bandelin (Berlin, Germany) for 40 min at 40  C. The extracts were separated from the solid matter by ltration, condensed to 1 ml, and analyzed for the contents of avonoids and polyphenols using high performance liquid chromatography (Heimler et al., 2006) with diode array detection (HPLC/DAD). 2.3. HPLC/DAD analysis of polyphenols and avonoids The HPLC/DAD was carried out with P580A LPG model liquid chromatograph, equipped with the Gina 50 model autosampler and the UVD340V DAD model diode array detector (Gynkotek/Dionex, Germering, Germany). The column (250 mm, 4.6 mm i.d.) was a Tosoh Biosep cartridge lled with TSK gel 5 mm; cat. # 08149, ODS-80 TM (Tosoh Corporation, Tokyo, Japan). The ow rate was 1 ml min1 and injection was via autosamples. The chromatographic column was maintained at 40  C. Sample volume for analysis was 50 ml. Each analysis lasted 50 min. Standards of polyphenols and avonoids in ethanol solutions (0.1 mg/ml) were vanillic acid, caffeic acid, p-coumaric acid, cinnamic acid, morin, hesperidin, neohesperdigo, quercetin, myricitin, apigenin, and campherol. The linear calibration plots were obtained by changing the injection volume of the individual standard solutions from 5 to 30 ml. The analyses were carried out with a changing gradient of the mobile phase composition (Table 1). 2.4. Determination of total polyphenols Lyophilized fruit samples were extracted from a 50-mg aliquot with 5 ml of 60% methanol/water with heating at 90  C for 3 h for free polyphenols (FP) and under the same conditions with 5 ml of 1.2 M HCl in 60% methanol/water for total polyphenols (TP) with some modications. The samples were cooled, diluted to 10 ml with methanol and centrifuged for 5 min at 4000g to remove solids (Vinson et al., 2001). For total polyphenol determination, the FolineCiocalteu method was used, and the measurement was performed at 765 nm with gallic acid as the standard. The results were expressed as mg gallic acid equivalents (GAE)/100 g FW (Singleton et al., 1999; Heimler, Vignolini, Dini, Vincieri, & Romani, 2006; Park et al., 2006). 2.5. Determination of avonoids The absorbance of avonoids (extracted with 5% NaNO2, 10% AlCl3$6H2O and 1 M NaOH) was measured at 510 nm
Table 1 The applied composition gradient of the binary ACN H2O mobile phase Time [min] Program 0e2 0e22 22e32 32e50 Constant composition Composition change Composition change Constant composition ACN [%] 5 Rise from 5 to 25 Rise from 25 to 55 55 H2O [%] 95 Drop 95 to 75 Drop from 75 to 45 45

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with the standards prepared similarly with known ()-catechin concentrations. The results were expressed as mg of catechin equivalents (CE)/100 g FW (Singleton et al., 1999). 2.6. Determination of total anthocyanins The total anthocyanins were measured by a pH differential method. Absorbance was measured in a Beckman spectrophotometer at 510 nm and at 700 nm in buffers at pH 1.0 and 4.5, using A [(A510 A700) pH 1.0 (A510 A700) pH 4.5]. Results were expressed as mg of cyanidin-3-glucoside equivalent (CGE)/100 g of FW (Cheng & Breen, 1991). 2.7. Determination of total avanols Total avanols were estimated using the p-dimethylaminocinnamaldehyde (DMACA) method. Methanolic extracts from durian samples (0.2 ml), diluted 1:100 with MeOH, were introduced into a 1.5-ml Eppendorf tube, and 1 ml of DMACA solution (0.1% in 1 N HCl in MeOH) was added. The absorbance at 640 nm was then read against a blank. The concentration of total avanols was estimated from the catechin calibration curve. Results were expressed as mg catechin equivalents (CE)/100 g FW (Arnous, Makris, & Kefalas, 2001). 2.8. UVevisible spectophotometric analysis and uorometry of polyphenols The spectra of methanol extracts in concentration of 0.02 mg/ml were measured on an Uvikon 930 (Bio-Teck-Kontron) and were recorded from 195 to 400 nm (Sarni-Manchado, Le Roux, Le Guerneve, Lozano, & Cheynier, 2000). Standards of caffeic acid and quercetin of 0.025 mM in methanol were used. Fluorescence measurements were done using a model FP6500, Jasco Spectrouorometer, serial N261332, Japan. Fluorescence emission spectra measurements for all fruit samples at a concentration of 0.02 mg/ml were taken at excitation wavelengths (nm) of 270 and emission of 290 recorded over the frequency range from the excitation wavelength to a wavelength of 500 nm. Standards of caffeic acid and quercetin of 0.01 mM in methanol were used (Gorinstein et al., 2001). 2.9. Determination of antioxidant capacities 2.9.1. b-Caroteneelinoleic acid assay The procedure was done according to Ferreira et al. (2006). A stock solution of b-carotene and linoleic acid was prepared by dissolving 0.5 mg of b-carotene in 1 ml of chloroform and adding 25 ml of linoleic acid with 200 mg of Tween 40. The chloroform was removed at 40  C under vacuum (evaporated), using a rotary evaporator (Rotavapor R-3000, Switzerland). Aerated water (100 ml) was added to the residue. To 2.5 ml of this mixture, 300 ml of durian acidic and non acidic methanol extracts were added. The samples were incubated in boiling water for 120 min together with two blanks, one containing

Fig. 1. UV absorption spectra of methanol extracts recorded from 195 to 400 nm. A, free polyphenols (0.02 mg/ml) of ripe (1), overripe (2) and mature (3) durian fruit. B, quercetin (0.025 mM). Fluorimetric spectra of methanol extracts taken at excitation wavelengths (nm) of 270 and emission of 290 recorded over the frequency range from the excitation wavelength to a wavelength of 500 nm. C, free polyphenols (0.02 mg/ml) of ripe (1), overripe (2) and mature (3) durian fruit. D, quercetin (0.01 mM).

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the antioxidant BHT and the other one without antioxidant. The absorbance was measured at 470 nm.

3. Results 3.1. UV and uorimetric spectra, total polyphenols, avonoids, anthocyanins and avanols

2.9.2. Ferric-reducing/antioxidant power (FRAP) The procedure was according to Szeto, Tomlinson, and Benzie (2002). FRAP reagent (2.5 ml of a 10 mM ferric-tripiridyltriazine solution in 40 mM HCl plus 2.5 ml of 20 mM FeCl3$H2O and 25 ml of 0.3 M acetate buffer, pH 3.6) of 900 ml was mixed with 90 ml of distilled water and 30 ml of durian samples or methanol as the appropriate reagent blank. The absorbance was measured at 595 nm.

2.9.3. Cupric-reducing antioxidant capacity (CUPRAC) The procedure was according to Apak et al. (2004). To the mixture of 1 ml of copper (II)-neocuproine and NH4Ac buffer solution, acidied and non acidied methanol extracts (or standard) solution (x, in ml) and H2O [(1.1 x) ml] were added to make the nal volume of 4.1 ml. The absorbance at 450 nm was recorded against a reagent blank.

2.10. Chemicals Trolox (6-hydroxy-2,5,7,8,-tetramethyl-chroman-2-carboxylic acid); BHA (butylated hydroxyanisole); b-carotene, FeCl3$6H2O; CuCl2$2H2O and neocuproine (2,9-dimethyl1,10-phenanthroline) were obtained from Sigma Chemical Co., St. Louis, MO, USA. 2,4,6-tripyridyl-s-triazine (TPTZ) was purchased from Fluka Chemie, Buchs, Switzerland. All reagents were of analytical grade. Deionized and distilled water were used throughout.

2.11. Statistical analyses The results of this investigation in vitro are means SD of three measurements. Differences between groups were tested by two-way ANOVA. In the assessment of the antioxidant potential, the Spearman correlation coefcient (R) was used. Linear regressions were also calculated. The p values of <0.05 were considered signicant.
Table 2 Bioactive compounds (100 g FW) in different durian samples Durian samples Ripe Overripe Mature Total polyphenols (mg GAE) 374.4 32.4a 298.5 24.4b 231.4 22.1c Free polyphenols (mg GAE) 45.4 4.6a 35.1 3.4b 27.3 2.9c

Polyphenols in the acidied and non acidied methanol extracts had absorption maxima in a range between 206 and 220 nm. The spectra were compared with the standards of caffeic acid and quercetin with the maximum between 206 and 215 nm, which indicated that avonoids were the predominant phenolic compounds (Ferreira da Silva, Lima, Quina, & Macuanita, 2004; Sarni-Manchado, Le Roux, Le Guerneve, Lozano, & Cheynier, 2000). The absorption UV maxima of acidied ripe durian were similar to caffeic acid (not shown). Non acidied samples (Fig. 1A) showed the following maxima (nm) and absorption units (AU): ripe with 209.1 and 0.812; overripe 213.2 and 0.678; mature 220.2 and 0.606, respectively, and were similar to quercetin with 206.7(Fig. 1B). The uorimetric data supported the UV results, where various contents of phenolic compounds were detected in the extracts, depending on the extraction solvent: acidied and non acidied and the stage of ripening. The following data of uorimetric measurements were obtained: for non acidied (free) polyphenols extracts: ripe durian with a peak of 306 nm and the absorption of 266.96 AU; mature 307 nm and 162.48 AU and the overripe 308 nm with the absorption of 163.28 AU (Fig. 1C). Quercetin, as well as caffeic acid showed the maximum of the peak at 304 nm (Fig. 1D). Total polyphenols (mg (GAE)/100 g) and avonoids (mg CE/100 g) in ripe durian (374.4 32.4 and 97.9 9.3) were signicantly higher ( p < 0.05) than in mature (231.4 22.1 and 57.3 6.1) and overripe (298.5 24.4 and 76.5 6.9, Table 2). The free polyphenols and avonoids were present at lower concentration than the hydrolyzed ones. Anthocyanins (mg of cyanidin-3-glucoside equivalent (CGE)/100 g FW) and avanols (mg of catechin equivalent (CE)/100 g FW) were signicantly higher in ripe durian (442.7 33.3 and 177.1 16.3) than in mature (388.5 41.1 and 155.4 15.7) and overripe (393.1 23.8 and 163.8 17.1), respectively. Identication of the compounds in the durian extracts was carried out through a comparison of their respective retention times (tR) with those of the applied standards (Table 3, Fig. 2), and their respective UV spectra with those of the standards. Quantication was calculated from the linear calibration plots. These were the absolute data detected by HPLC analysis on

Total avonoids (mg CE) 97.9 9.3a 76.5 6.9b 57.3 6.1c

Free avonoids (mg CE) 23.9 2.4a 19.3 1.9b 14.7 1.5c

Anthocyanins (mg CGE) 442.7 33.3a 393.1 23.8b 388.5 41.1b

Flavanols (mg CE) 177.1 16.3a 163.8 17.1b 155.4 15.7c

Values are means SD of three measurements. Means in columns without superscript letters in common differ signicantly (P < 0.05). Abbreviations: FW, fresh weight; GAE, gallic acid equivalent; CE, catechin equivalent; CGE, cyanidin-3-glucoside equivalent.

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Table 3 Compounds identied by HPLC/DAD in the durian extracts, their respective retention times, tR [min], and concentrations calculated in mg mg1 of the lyophilized dry matter Compounds Overripe tR Vanillic acid Caffeic acid p-Coumaric acid Cinnamic acid Morin Hesperidin Neohesperdigo Quercetin Myricitin Apigenin Campherol 2.80 3.51 e 9.37 23.18 e 28.59 e 32.04 33.92 34.27 C 0.032 0.012 e 0.038 0.062 e 0.029 e 0.016 0.138 0.282 Ripe tR 2.80 3.50 6.58 9.43 23.13 e e 31.65 32.04 33.92 34.26 C 0.011 0.017 0.023 0.029 0.024 e e 0.053 0.014 0.027 0.095 Mature tR 2.80 e 6.70 9.47 24.01 28.12 e e 32.05 e 34.26 C 0.016 e 0.032 0.032 0.006 0.011 e e 0.057 e 0.059

Values are means of three measurements.

the weight of lyophilized sample (Table 3). Caffeic acid (mg/ 100 g) was signicantly higher in ripe 490 vs. 360 than in overripe durian and was not detected in mature fruit. Quercetin of 1200 mg/100 g was estimated only in ripe durian sample (Table 4).

4. Discussion Fruits have long been regarded as having considerable health benets, due to their main antioxidant compounds, of which phenolics are the most abundant (Gorinstein et al., 2006; Park et al., 2006; Sarni-Manchado, Le Roux, Le Guerneve, Lozano, & Cheynier, 2000). A large screening study of the antioxidant capacity of methanol extracts of fruits reported that these fruits contain different quantities of antioxidant compounds and have different levels of antioxidant capacity (Halvorsen et al., 2002). Durian fruit was not studied yet in such a way. Therefore, this fruit at different stages of its ripening was investigated in this study as a fruit diet and as an additive to functional foods for prevention of cardiovascular and other diseases as traditional fruits (Miller, Liebowitz, & Newby, 2004). Free radicals require the ability to measure them and the oxidative damage that they cause (Halliwell & Whiteman, 2004); therefore, in this study the radical scavenging assays were carried out to show the ability of the durian extracts to scavenge free radicals in vitro (expressed as TEAC value) by CUPRAC and FRAP. The antioxidant capacity is mainly derived from the alcohol soluble antioxidants and has a high correlation coefcient with polyphenols (0.97), which corresponds with others (CaballeroTable 4 Compounds identied by HPLC/DAD in durian extracts and concentrations calculated in mg/100 g FW Compound Overripe 970 360 e 720 1900 e 880 e 480 4200 8500 Ripe 250 490 530 660 550 e e 1200 320 620 2200 Mature 300 e 600 600 110 200 e e 1060 e 1100

3.2. Antioxidant capacity The ripe durian had the highest amounts of antioxidant capacity and bioactive compounds (Table 5). FRAP absorption measurements (Fig. 3 A and B) showed that durian at the ripe stage had higher antioxidant activity than the other two samples. A very good correlation was observed between the antioxidant capacities determined by FRAP and CUPRAC (Fig. 4, A and B) and the total polyphenols (R2 is 0.972 and 0.891, respectively). The correlation coefcients between the antioxidant capacity determined by FRAP and CUPRAC and for avonoids (Fig. 4 A and B) were lower than for total polyphenols (R2 is 0.865 and 0.711, respectively).
100

mAV

50 3 4 5 2 0 0 10 20 30 40 50 6 7 8

time [min]
Fig. 2. The chromatogram obtained from the Durian overripe extract. The identied compounds: (1) vanillic acid; (2) caffeic acid; (3) cinnamic acid; (4) morin; (5) neohesperdigo; (6) myricetin; (7) apigenin; and (8) campherol.

Vanillic acid Caffeic acid p-Coumaric acid Cinnamic acid Morin Hesperidin Neohesperdigo Quercetin Myricitin Apigenin Campherol

Values are means of three measurements.

P. Arancibia-Avila et al. / LWT - Food Science and Technology 41 (2008) 2118e2125 Table 5 Antioxidant activity in different durian samples (in 100 g FW) Durian samples FRAP, reducing/ antioxidant power (mMTE) 270.4 27.2a 257.5 24.8a 217.4 20.9b CUPRAC, cupric-reducing antioxidant capacity (mMTE) 1112.7 83.4a 1091.2 72.5a 1019.8 68.5a b-Carotene, % inhibition

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110 100

A
mgGAE/100g FW

400 380 360 340 320 300 280 260 240 220 200 210 220 230 240 y = 0.6694x - 90.247 R2 = 0.8652 y = 2.8125x - 395.63 R2 = 0.972

90 80 70 60 50 40 280

mgCE/100g FW

Ripe Overripe Mature

76.8 6.8a 70.6 6.3b 64.3 5.3c

Values are means SD of 3 measurements. Means in columns without superscript letters in common differ signicantly (P < 0.05). Abbreviations: FW, fresh weight; TE, trolox equivalent.

250

260

270

George et al., 2002). The UV and uorimetric spectra of acidied and non acidied extracts were close to each other and showed higher intensity of ripe durian in comparison with two other samples. Such interpretation of our results

MTE/100g FW

B
mgGAE/100g FW

400 380 360

110 y = 0.9014x - 684.03 R2 = 0.8906 100 90 80 70 y = 0.1985x - 141.88 R2 = 0.7114 60 50 40 1250

A
Absorbance, 593 nm

0.30 0.25

mgCE/100g FW

340 320 300 280 260 240

0.20 0.15

220 200 950 1000 1050 1100 1150 1200

0.10 0.05

MTE/100g FW
Fig. 4. Correlation between measures of antioxidant capacities (AC) and total phenolics (TPOL). A, (A) ACFRAP (mMTE/100 g FW, X) and TPOL (mg GAE/100 g, Y1), (-) ACFRAP (mMTE/100 g, X) and TFLAV (mg CE/ 100 g FW, Y2). B, (>) ACCUPRAC (mMTE/100 g, X) and TPOL (mg GAE/100 g, Y1), (,) ACCUPRAC (mMTE/100 g, X) and TFLAV (mg CE/ 100 g, Y2). Abbreviations: FRAP, Ferric-reducing/antioxidant power; CUPRAC, Cupric-reducing antioxidant capacity; TPOL, Total polyphenols; TFLAV, Total avonoids.

0.00 0 1 2 3 4 5

Time (min)

B
Absorbance, 593 nm

0.6 0.5 0.4 0.3 0.2 0.1 0 0 5 10 15 20 25

Total phenols (mg/ml)

Fig. 3. FRAP (Ferric-reducing/antioxidant power) measurements of durian extracts with two variables (different concentrations and different periods of time): A, total polyphenol extracts of durian cultivar Mon Thong at different stages of ripening: (A) M, mature; (-) O, overripe; (:) R, ripe at constant concentration (10 mg/ml) and with the change of time (1, 2, 3, and 4 min). Standards (30 mg/ml), (3) NAR, (*) FIC, () QUER, naringin, citin and quercetin, respectively; (B) GA, gallic acid, 10 mg/ml; B, total polyphenol extracts from durian cultivar Mon Thong at different stages of ripening: (A) C, mature; (-) D, overripe; () G, ripe at constant time (4 min) with the change of extracts concentration (5, 10, 15 and 20 mg/ml).

corresponds with Ferreira da Silva et al. (2004). Another possibility for explanation of the high antioxidant capacity can be the total activity of all phenolic acids (Nilsson et al., 2005). Our results were similar to those reported for strawberry and banana (Mahattanatawee et al., 2006). It was interesting to compare between different stages of ripening; mostly our data correspond with guava and mango. Ripe and green mangos and papayas slightly differ in the total polyphenols from our results (Mahattanatawee et al., 2006). The phenolic content and the % of inhibition by b-carotene of the investigated fruits corresponded with wild mulberry and commercial frozen pulp of mulberry as shown by Hassimotto, Genovase, and Lajolo (2005). The numbers of total polyphenols in durian corresponded with the data of Wu et al. (2005) for plums and bananas. Our results of Mon Thong ripe durian polyphenol content are similar to plums and mature fruits correspond with strawberries (Chun et al., 2005). The amount of avonoids and the antioxidant activity for Mon Thong cultivar in ripe stage was lower than in plums and mature fruit was equal with strawberries.

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P. Arancibia-Avila et al. / LWT - Food Science and Technology 41 (2008) 2118e2125 Arnous, A., Makris, D. P., & Kefalas, P. (2001). Effect of principal polyphenolic components in relation to antioxidant characteristics of aged red wines. Journal of Agricultural and Food Chemistry, 49, 5736e5742. Caballero-George, C., Vanderheyden, P. M. L., De Bruyne, T., Shahat, A. A., Van den Heuvel, H., & Solis, P. N., et al. (2002). In vitro inhibition of [H3]-angiotensin II binding on the human AT(1) receptor by proanthocyanidins from Guazuma ulmifolia bark. Planta Medica, 68, 1066e1071. Cheng, G. W., & Breen, P. J. (1991). Activity of phenylalanine ammonia-lyase (PAL) and concentrations of anthocyanins and phenolics in developing strawberry fruit. Journal of the American Society of Horticultural Science, 116, 865e869. Chun, O. K., Kim, D.-O., Smith, N., Schroeder, D., Han, J. T., & Lee, C. Y. (2005). Daily consumption of phenolics and total antioxidant capacity from fruit and vegetables in the American diet. Journal of the Science of Food and Agriculture, 85, 1715e1724. Dauchet, L., Amouyel, P., Hercberg, S., & Dallongeville, J. (2006). Fruit and vegetable consumption and risk of coronary heart disease: a meta-analysis of cohort studies. Journal of Nutrition, 136, 2588e2593. Erkkilae, A. T., Herrington, D. M., Mozaffarian, D., & Lichtenstein, A. H. (2005). Cereal ber and whole-grain intake are associated with reduced progression of coronary-artery atherosclerosis in postmenopausal women with coronary-artery disease. American Heart Journal, 150, 94e101. Ferreira, A., Proenca, C., Serralheiro, M. L. M., & Araujo, M. E. M. (2006). The in vitro screening for acetylcholinesterase inhibition and antioxidant activity of medicinal plants from Portugal. Journal of Ethnopharmacology, 108, 31e37. Ferreira da Silva, P., Lima, J. C., Quina, F. H., & Macuanita, A. L. (2004). Excited-state electron transfer in anthocyanins and related avylium salts. Journal of Physical Chemistry A, 108, 10133e10140. Gorinstein, S., Caspi, A., Libman, I., Lerner, H.-Z., Huang, D., & Leontowich, H., et al. (2006). Red grapefruit positively inuence serum lipids level in patients suffering from coronary atherosclerosis: studies in vitro and in humans. Journal of Agricultural and Food Chemistry, 54, 1887e1892. Gorinstein, S., Delgado-Licon, E., Pawelzik, E., Permadym, H., H,Weisz, M., & Trakhtenberg, S. (2001). Characterization of soluble amaranth and soybean protein proteins based on uorescence, hydrophobicity, electrophoresis, amino acid analysis, circular dichronism and differential scanning calorimetry measurements. Journal of Agricultural and Food Chemistry, 49, 5595e5601. Halliwell, B., & Whiteman, M. (2004). Measuring reactive species and oxidative damage in vivo and in cell culture: how should you do it and what do the results mean? British Journal of Pharmacology, 142, 231e255. Halvorsen, B. L., Holte, K., Myhrstad, M. C. W., Barikmo, I., Hvattum, E., & Remberg, S. F., et al. (2002). A systematic screening of total antioxidants in dietary plants. Journal of Nutrition, 132, 461e471. Hassimotto, N. M. A., Genovase, M. I., & Lajolo, F. M. (2005). Antioxidant activity of dietary fruits, vegetables, and commercial frozen fruit pulps. Journal of Agricultural and Food Chemistry, 53, 2928e2935. Heimler, D., Vignolini, P., Dini, M. G., Vincieri, F. F., & Romani, A. (2006). Antiradical activity and polyphenol composition of local Brassicaceae edible varieties. Food Chemistry, 99, 464e469. Jung, H.-A., Su, B.-N., Keller, W. J., Mehta, R. G., & Kinghorn, A. D. (2006). Antioxidant xanthones from the pericarp of Garcinia mangostana (Mangosteen). Journal of Agricultural and Food Chemistry, 54, 2077e2082. Ketsa, S., & Daengkanit, T. (1998). Physiological changes during postharvest ripening of durian fruit (Durio zibethinus Murray). Journal of Horticulture Science and Biotechnology, 73, 575e577. Koebnick, C., Garcia, A. L., Dagnelie, P. C., Strassner, C., Lindemans, J., & Katz, N., et al. (2005). Long-term consumption of a raw food diet is associated with favorable serum LDL cholesterol and triglycerides but also with elevated plasma homocysteine and low serum HDL cholesterol in humans. Journal of Nutrition, 135, 2372e2378. Lairon, D., Arnault, N., Bertrais, S., Planells, R., Clero, E., & Hercberg, S., et al. (2005). Dietary ber intake and risk factors for cardiovascular disease in French adults. American Journal of Clinical Nutrition, 82, 1185e1194. Mahattanatawee, K., Manthey, J. A., Luzio, G., Talcott, S. T., Goodner, K., & Baldwin, E. A. (2006). Total antioxidant and ber content of select Florida-grown tropical fruits. Journal of Agricultural and Food Chemistry, 54, 7355e7363.

The comparison of the cited and the present data was done using different solvents for the extraction of bioactive compounds: methanolic extracts of durian samples (0.2 mg/ml) and the decoction extracts (0.1 mg/ml). Our results of inhibition by b-carotene assay for ripe, overripe and mature durians correspond with Melissa ofcinalis and Lavandula pedunculata (Ferreira et al., 2006). The present results can be compared as well with Hassimotto et al. (2005), where the samples investigated by b-carotene bleaching system gave inhibition values >70%. BHT (78.1 1.6), quercetin (48.8 4.7) and rutin (13.6 1.4) were measured in the present report and compared with the corresponding data of 77.6 0.4, 49.2 3.4 and 12.7 2.3 obtained by Hassimotto et al. (2005). The durian ripe sample can be compared with the data of BHT (50 mM). The antioxidant capacity of mature durian was between BHT and quercetin. The DPPH (1,1-diphenyl-2-picrylhydrazyl, radical scavenging activity) antioxidant activity and ORAC (oxygen radical absorbance capacity) determined in different stages of maturity showed that the ripe papaya showed 2.19 and 2.04 times higher values than the green ones (29.7; 2.6) (Mahattanatawee et al., 2006). In this experiment, the antioxidant activity determined by FRAP, CUPRAC and b-carotene showed slightly lower relations between the ripe and mature samples such as 1.24; 1.09 and 1.19, respectively. The FRAP values of the durian samples corresponded with apple and banana, according to Halvorsen et al. (2002), and were lower than banana and similar to mango, according to Nilsson et al. (2005). In conclusion, (1) it is preferable to consume ripe Mon Thong durian, which has higher content of bioactive compounds and possesses higher antioxidant capacity than the mature and overripe samples and (2) according to the results of the present and our previous investigations, the antioxidant capacity of tested tropical fruits was in decreasing order: ripe Mon Thong durian > snake fruit > mangosteen > lichi > guava > mango. Acknowledgements The authors are thankful to Dr. Elena Katrich (Hebrew University of Jerusalem, School of Pharmacy, Israel) for her technical assistance in determination of antioxidant activity, Dr. Ratiporn Haruenkit and Dr. Sumitra Poovarodom (King Mongkuts Institute of Technology Ladkrabang, Ladkrabang, Bangkok, Thailand) for supplying the durian samples. This work was partly supported by the Food Industrial Technology Innovation Center at Mokpo National University. References
Apak, R., Guclu, K., Ozyurek, M., & Karademir, S. E. (2004). Novel total antioxidant capacity index for dietary polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. Journal of Agricultural and Food Chemistry, 52, 7970e7981.

P. Arancibia-Avila et al. / LWT - Food Science and Technology 41 (2008) 2118e2125 Miller, K. L., Liebowitz, R. S., & Newby, L. K. (2004). Complementary and alternative medicine in cardiovascular disease: a review of biologically based approaches. Americam Heart Journal, 147, 401e411. Nilsson, J., Pillai, D., Onning, G., Persson, C., Nilsson, A., & Akesson, B. (2005). Comparison of the 2,2-azinobis-3-ethylbenzotiazoline-6-sulfonic acid (ABTS) and ferric-reducing antioxidant power (FRAP) methods to asses the total antioxidant capacity in extracts of fruit and vegetables. Molecular Nutrition Food Research, 49, 239e246. Ou, B., Huang, D., Hampsch-Woodill, M., Flanagan, J., & Deemer, E. (2002). Analysis of antioxidant activities of common vegetables employing oxygen radical absorbance capacity (ORAC) and ferric-reducing antioxidant power (FRAP) assays: a comparative study. Journal of Agricultural and Food Chemistry, 50, 3122e3128. Park, Y.-S., Jung, S.-T., Kang, S.-G., Drzewiecki, J., Namiesnik, J., & Haruenkit, R., et al. (2006). In vitro studies of polyphenols, antioxidants and other dietary indices in kiwifruit (Actinidia deliciosa). International Journal of Food Sciences and Nutrition, 57, 107e122. Sarni-Manchado, P., Le Roux, E., Le Guerneve, C., Lozano, Y., & Cheynier, V. (2000). Phenolic composition of litchi fruit pericarp. Journal of Agricultural and Food Chemistry, 48, 5995e6002.

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Singleton, V. L., Orthofer, R., & Lamuela-Raventos, R. M. (1999). Analysis of total phenols and other oxidation substrates and antioxidants by means of FolineCiocalteu reagent. Methods Enzymology, 299, 152e 178. Szeto, Y. T., Tomlinson, B., & Benzie, I. F. F. (2002). Total antioxidant and ascorbic acid content of fresh fruits and vegetables: implications for dietary planning and food preservation. British Journal of Nutrition, 87, 55e59. Vinson, J. A., Su, X., Zubic, L., & Bose, P. (2001). Phenol antioxidant quantity and quality of foods: fruits. Journal of Agricultural and Food Chemistry, 49, 5315e5321. Wu, X., Beecher, G. R., Holden, J. M., Haytowitz, D. B., Gebhardt, S. E., & Prior, R. L. (2005). Lipophilic and hydrophilic antioxidant capacities of common foods in the United States. Journal of Agricultural and Food Chemistry, 52, 4026e4037. Yaacob, O., & Subhadrabandhu, S. (1995). The production of economic fruits in South-East Asia. Kuala Lumpur: Oxford University Press. Zhang, X., Koo, J., & Eun, J.-B. (2006). Antioxidant activities of methanol extracts and phenolic compounds in Asian pear at different stages of maturity. Food Science and Biotechnology, 15, 44e50.