ORGANIC AND CONVENTIONAL KIWIFRUIT: MYTHS VS.

REALITY, ANTIOXIDANT, QUENCHING, ANTIPROLIFERATIVE AND HEALTH EFFECTS

Yong Seo Park1, Myang Hee Im2, Hanna Leontowicz3, Maria Leontowicz3, Milan Suhaj4, Elena Katrich5, Moshe Weisz5, Zeev Tashma5, Shela Gorinstein5#
1Department

of Horticultural Science, Mokpo National University, Jeonnam, South Korea; 2Regional Crop Research Institute, Mokpo National University, Jeonnam, South Korea; 3Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences (SGGW), Warsaw, Poland; 4Food Research Institute, Bratislava, Slovakia; 5The Institute for Drug Research, The Hebrew University of Jerusalem, Faculty of Medicine, School of Pharmacy, Jerusalem, Israel

#This research is dedicated to the memory of my dear brother Prof. Simon Trakhtenberg, who encouraged and supported me and our research group during all his life.

SG-073_120311101021 #45-56 RT: 0.40-0.50 AV: 12 SB: 13 0.16-0.27 NL: 7.79E5 T: - p ESI Q1MS [100.070-900.000] 100 95 90 85 80 75 390.64

Relative Abundance

1. Background
Fruits with high content of bioactive compounds are effective in prevention and treatment of atherosclerosis (1-4). In this investigation the bioactivity of conventionally and organically grown kiwifruits cultivars Hayward (KHaC', KHaO') and 'Bidan' (KBiC', KBiO') were studied.
Relative Abundance

70 65

SG-071_120311095808 #77-105 RT: 0.69-0.95 AV: 29 SB: 11 0.13-0.22 NL: 1.57E6 60 T: - p ESI Q1MS [100.070-900.000] 55 110.85 100
50 45 40 35 30 25 20 15 10 5

95 90 85 80 75 70 65 60 55 50 45 40 35 30 25 20 15 10 5 0 100

2. Materials and methods

Methanol (MeOH) and ethyl acetate (EtOAc) extracts of conventionally and organically grown in Korea (Fig.1) kiwifruits cultivars Hayward (KHaC', KHaO') and 'Bidan (KBiC', KBiO') were investigated. FTIR, 3D-FL spectroscopy, ESI-MS, DSC and radical scavenging assays were used for characterization of bioactive compounds and the levels of their antioxidant activities (1-4). Fig. 1. Kiwifruit orchard Heanam County, Jeonnam province,
Korea, 2011 (S.Gorinstein, Y.S. Park)

5 154.81 SG-072_120311100448 #51-137 RT: 0.46-1.24 AV: 87 SB: 16 0.12-0.26 NL: 9.22E5 T: - p ESI Q1MS [100.070-900.000] 0 100 150 110.92 100

95 90 85 80 75 70 65

Relative Abundance

60 55 50 45 40 35 30 25 20 15 10 5

0 100

The main peaks (m/z) with Relative Abundance (RA, %), using different extraction procedures MeOH and EtOAc, for KBiCMeOH' and KBiOMeOH' (Fig. A 4A and 4B) were at: 111(100, 100), 129(15, 15), 175(55, 55), 191(75, 75), 391(40, 80); for KBiCEtOAc' and KBiOEtOAc' (inserts in 4A and 4B): 111(12, 28), 217(45, 40), 311(30, 28), 325 (50,33), 339(43,30), 391(100,100). The main m/z peaks for KHaCMeOH' and KHaOMeOH' were at: 111(38, 43), B 191(100, 100) and 391(47, 90); KHaCEtOAc' and KHaOEtOAc' - 111(12, 12), 217(22, 22); 311 (15, 12), 325(20, 18), 339(12, 12), 391(100, 100). The difference between conventional and organic Fig. 4. ESI-MS spectra of (A): 'Hayward' and 'Bidan' cultivars was only methanol fraction (MeOH) of KBiC with an insert of ethyl acetate (EtOAc) in RA of peaks, but between the cultivars fraction of KBiC; (B), MeOH was also in the shift of the peaks.
324.84 216.62 338.91 310.91

190.86

111.13

132.69

352.98

408.84

380.98

158.73

309.09

115.19

276.96

154.95 150

173.78

199.12 200

242.87

265.27

379.02

432.99 440.97

301.11 300

469.04

483.11 484.93 513.00 500

537.01 557.10 550

579.15

591.12 600

0 100

250

350 m/z

400

450

174.69

390.78

222.71

262.89

340.87

128.77

438.80

404.85

172.87

436.77

154.67

216.76

228.80

276.61

318.82 292.71 310.70

380.77

352.77

424.73

442.79 450

480.73

502.78 500

533.02

570.82

592.73

150

200

250

300

350 m/z

400

550

600

SG-074_120311101258 #57-66 RT: 0.51-0.59 AV: 10 SB: 14 0.16-0.28 NL: 1.33E6 T: - p ESI Q1MS [100.070-900.000] 100 95 90 85 80 75 70 65 60 55 50 45 40 35 30 25 20 15 10

390.71

Relative Abundance

216.69

110.99

325.05 338.77

311.05

264.71

292.99

352.91

132.83

380.98

129.19

158.87

408.77 415.14

440.97

172.59

212.98

231.04

258.97 250

277.10

378.60

468.90 483.04

513.07 528.89

555.07

576.98 596.93 600

200

300

350 m/z

400

450

500

550

390.78

190.86

174.76

404.78

222.71

262.61

340.80

128.77

438.80 436.77

172.80

216.62

146.83

202.76

232.93

292.78

318.75 332.82

352.63

380.63

424.87

442.93 450

476.88

502.99 500

532.88

564.94

592.80

150

200

250

300

350 m/z

400

550

600

3. Results
The significantly highest levels of bioactive compounds were estimated in methanol extract of KBiO' (P < 0.05) than in other investigated kiwifruit samples, but the difference between the organic and conventional kiwifruit was not significant (Fig. 2).
A B A

fraction of KBiO with an insert of EtOAc fraction of KBiO in negative ion mode.

Fig. 5. DSC curves of EtOAc fractions of (A): KHaO (a), KHaC (b), KBiO (c), KBiC (d); Fig. 2. A, B, three-dimensional fluorescence (3D-FL) spectrum of methanol extracts of
conventional and organic kiwifruits Bidan (0.25 mg/mL) MeOH fractions (B): KHaO (a), KHaC (b), KBiO (c), KBiC (d); MeOH extracts (C): KHaC (a), KHaO (b), KBiC (c), KBiO (d). The pans were heated in the calorimeter at 10 oC min over the range 25-200 oC.

Endothermic peaks were at the same temperature [EtOAc and MeOH fractions (Fig.5A and 5B)] with nearly 3-fold and 14-fold more heat was required to soften the KHaC that the KHaO. MeOH extract 2 -fold more heat required for KHaO (Fig 5C). The endothermic peaks for KBiC and KBiO were at the same temperature with the same number of peaks for EtOAc, but the enthalpy of transition was different and as in case of 'Hayward' 1.8-fold more heat required for conventional. In MeOH fractions and extracts the temperature peaks were similar, but 3.3-fold and 5.2-fold more heat required for organic 'Bidan'.

Fig. 3. Sun ray icon plot of organically (O) and conventionally (C) grown Hayward kiwifruit
treated with ethylene (0,6,12,18,24 hrs) were compared by the examined variables (fruit firmness, sensory value, soluble solid content, total tartaric acidity , vitamin C, antioxidant activity, electron donating ability , nitrite scavenging activity , angiotension-1 converting enzyme , total phenolics). Each ray represents a different variable; the middle of the ray comprises the mean value of the variable. Values for each parameter are connected by a cord. This plot clearly demonstrates the found differences between organic and conventional kiwifruit [Principal Component Analysis (PCA), Fig.3].
120
120

4. Discussion
The DSC thermograms of the phenolic extracts showed only one endothermic peak at different temperatures, depending on investigated samples and year of collection. The kiwifruit extracts decreased the proliferation of both Calu-6 and SNU-601 for human pulmonary carcinoma and gastric carcinoma cells, and the effect was concentration dependent. The proliferativity for concentrations of 1000g/ml for KBiO' methanol extract was lower than for other investigated samples. The interaction between drugs and human serum albumin plays an important role in the distribution and metabolism of drugs. The complexation reaction between flavonoids, and kiwifruit extracts, and bovine serum albumin (BSA) showed that kiwifruit polyphenol extracts have strong ability to quench the intrinsic fluorescence of BSA and comparable with quercetin (1-4).

A
100 80 60 40 20

Calu-6 SNU-601
100

Calu-6 SNU-601

Cell viability, %

Cell viability, %

80 60 40 20 0

.0
10 30 100 300 1000

10

30

100

300

1000

Extract concentration, m g/mL

120
120

Extract concentration, m g/mL

C
100 80

Calu-6 SNU-601

D
100

Calu-6 SNU-601

Cell viability, %

Cell viability, %

80

60 40 20 0 10 30 100 300 1000

60 40 20 0 10 30 100 300 1000

The proliferativity (%) at 1000 g/mL (Fig. 6A, B) for 'KHaO' on Calu-6 is 54.54%, and on SNU-601 is 60.08%, 'KHaC' for Calu-6 (56.23%) and SNU-601 (61.56%); for 'KBiO' on Calu-6 is 48.84%, on SNU-601 is 53.36% (Fig. 6C, D), 'KBiC' for Calu-6 (47.25%) and SNU-601 (52.28%). Our results show that antioxidant activity of the studied samples correlates with their antiproliferative activity. Significantly different values between the organic and conventional samples were not found. Fig. 6. Cell viability (% of Control) of human
cancer cells of the Calu-6 and Snu-601 lines in the presence of MeOH extracts: A, KHaC; B, KHaO; C, KBiC; D, KBiO.

5. Conclusion
Relatively high contents of bioactive compounds, positive antioxidant and antiproliferative properties of two cultivars of kiwifruit justify their use as a source of valuable antioxidants.

6. References
1. Gorinstein et al., 2010. Phytochemical Analysis, 21, 355-362. 2. Park et al., 2010. Plant Foods for Human Nutrition, 65, 186191. 3. Im et al., 2012. International Journal of Food Properties, 15, 4959. 4. Leontowicz et al., 2008. Food and Chemical Toxicology, 46, 581589. 5. Our publications- http://www.bashanfoundation.org/shela/shelapub.html

Extract concentration, m g/mL

Extract concentration, m g/mL