华 北 农 学 报 2013,28 ( 2 ) : 1 44-1 4 9 300072 OH DP- PH O 2 - FRAP 4 3. 19 ± 0. 53 mg /g 0. 26 ± 0. 07 mg /g OH DPPH O 2-2 g /ml OH DPPH 84. 5% 82. 2% 1 g /ml O 2-90. 7% DPPH O 2 - OH S663. 4 A 1000-7091 2013 02-0144 - 06 Investigation on the Anti-oxidative Activity of Kiwifruits Pulp Extracts WANG Hua CAO Jing ZHAI Li-juan XUE Zhao-hui Department of Food Science College of Chemical Engineering and Technology Tianjin University Tianjin 300072 China Abstract By measuring systematically in simulated chemical systems through the ways of scavenging DPPH ABTS and superoxide anion radicals and FRAP method the antioxidant activity of pulp with flash extraction and the content of the total phenol and total flavonoids of the extract were measured. The results showed that the content of total phenol and total flavonoids of the extract were 3. 19 ± 0. 53 mg /g and 0. 26 ± 0. 07 mg /g respectively. The research in simulated chemical systems showed that the acetone-extracting from kiwifruits had great scavenging effect against OH DPPH and O 2 - in the experimental range. When the concentrate of sample was 1 g /ml the scavenging rate to superoxide anion radicals was up to 90. 7%. And when the concentrate was 2 g /ml the scavenging rate to OH and DPPH was 84. 5% and 82. 2% respectively. With the increasing concentration the scavenging effect enhanced which suggested the presence of dose-effect relations. Beside the FRAP value of also increased with concentration which had excellent significant correlation with its DPPH and O 2 - radical-scavenging activity significant correlation with its OH radical-scavenging activity. The results provide the theory basic for kiwifruits exploitation. Key words Kiwifruits Acetone-extracting Total phenol Flavonoids Antioxidant activity 1 C E 2012-12 - 03 1986-1973 -
2 期 王 华 等 : 猕 猴 桃 果 肉 提 取 物 抗 氧 化 活 性 研 究 145 2 1. 1 1. 4 1. 2 DPPH 1 1-mg 70% 50-2- TBA ml 70% 0. 52 K 2 S 2 O 8 TPTZ 2 4 6- Sigma Folin- 70% 3. 00 ml 5% 0. 50 1. 00 1. 50 2. 00 2. 50 3. 00 ml 25 ml 0. 75 ml 5 min 10% TDL-5 7230G 0. 5 ml 6 min 5% RE52CS 4 ml 15 min 510 JHBE-50S nm UV1102 1. 3 Wolfe 7 0. 5 ml 6 min 5% 500 g 1 2 80% 4 ml 25 15 min 5 min 180 V 1 min510 nm 10 ml 80% 10 2 1. 4. 3 OH 45 0. 4 ml 50 mmol /L 10% 1 d ph 7. 5 0. 1 ml - 56-40 1. 4. 1 Foiln-Ciocalte 8 0. 05 g 20 3 100 ml 500 μg /ml 500 μg /ml 50 100 200 300 4 400 μg /ml 5 500 μl Folin- 6 2. 5 ml 7. 5% 2 ml 10 ml 45 15 min 10 s 4 000 r /min 20 min765 nm 500 μl Folin- 2. 5 ml 7. 5% Na 2 CO 3 2 ml 10 1 ml 45 15 min 10 s 4 000 r /min 20 min765 nm 9 1. 4. 2 26. 0 mg /ml 0. 00 2 ml 5% 0. 75 ml 5 min 10% 0. 1 ml 1. 04 mmol /L EDTA 0. l ml
146 华 北 农 学 报 28 卷 10 mmol /L H 2 O 2 0. 1 ml 2 mmol /L 0. l 0. 1 ml 3 ml FRAP ml 60 mmol /L DR DR 0. 3 ml 5 min 0. 1 ml 1 mmol /L FeCl 3 593 nm 3 1. 0 ml 37 1 h 1 ml 25% V /V 1 ml 1% TBA 15 2. 1 min 532 nm 500 g 3 ml 49. 85 g 9. 97% 2. 2 = A 0 - A 1 - A 2 /A 0 100% A 0 A 1 A 2 y = 0. 006 7x + 0. 034 5 r 2 = 0. 999 2 50 ~ 400 μg /ml 1. 4. 4 DPPH 11 1 ml 1 ml 0. 1 mmol /L DPPH 20 min 517 nm A i 6 500 14 DPPH 517 nm A j 517 nm A 0 = 1 - A i - A j /A 0 100% A 0 A i A j 1. 4. 5 O 2-12 0. 2 ml 4. 6 ml 0. 1 mol /L Tris-HCl ph 8. 2 25 10 min 0. 1 ml 3 mmol /L 25 325 nm 30 s 4 min 3 0. 2 ml 1 V Fig. 1 Standard curve for gallic acid 2. 3 = V - V /V 100% V V 11. 315x - 0. 002 r 2 = 0. 999 6 50 ~ 400 μg /ml 1. 4. 6 FRAP 13 0. 1 ml 0. 1 0. 2 0. 4 0. 6 0. 8 1. 0 mmol /L FeSO 4 3 ml FRAP 25 ml 300 2 1 3. 19 ± 0. 53 mg /g 15 2 y = 16-17 mmol /L ph 3. 6 2. 5 ml 10 Whang 18 mmol /L TPTZ 2. 5 ml 20 mmol /L Webby 19 0. 3 ml 5 min 593 nm 0. 26 ± 0. 07 mg /g FRAP FeSO 4 2. 4 OH OH
2 期 王 华 等 : 猕 猴 桃 果 肉 提 取 物 抗 氧 化 活 性 研 究 147 3 OH 0. 05 ~ 2 g /ml OH 0. 01 ~ 0. 5 g /ml OH 0. 5 ~ 2 g /ml OH 2 g /ml Fig. 4 OH 84. 5% DPPH radical-scavenging activities of 2. 6 O 2 - Fig. 2 2 Standard curve for Rutin 4 DPPH O 2 - O 2 - NBT Diformazan 560 nm 21 5 O 2-0. 025 ~ 1 g /ml O 2-0. 025 ~ 0. 5 g /ml O 2-0. 5 ~ 1 g /ml O 2-1 g /ml O 2-90. 7% 3 OH Fig. 3 OH-scavenging activities of 2. 5 DPPH DPPH 517 nm 5 O 2 - Fig. 5 O 2 - -scavenging activities of 2. 7 FRAP 4 0. 200 6x + 0. 046 2 r 2 = 0. 996 9 FeSO 4 0. 1 ~ DPPH 1. 0 mmol /L 0. 05 ~ 2 g /ml DPPH Fe 3 + Fe 2 + Fe 2 + 2 4 6-2 g /ml 2- TPTZ DPPH 82. 2% 20 6 FeSO 4 y = 593 nm
148 华 北 农 学 报 28 卷 FRAP 22 21 6 FeSO 4 Fig. 6 Standard curve for FeSO 4 7 4 0. 025 ~ 1 g /ml OH DPPH O 2-1 g /ml 7 FRAP Fig. 7 FRAP value of 2. 8 1 DPPH O 2 - OH 1 Tab. 1 Correlations of anti-oxidative activities of Correlation OH DPPH O 2 - FRAP coefficient OH 1 DPPH 0. 988 ** 1 O 2-0. 963 ** 0. 981 ** 1 FRAP 0. 841 * 0. 902 ** 0. 931 ** 1 * P < 0. 05 ** P < 0. 01 Note * and ** shows significant difference at 0. 05 and 0. 01 level respectively. 3 23 3. 19 ± 0. 53 mg /g 0. 26 ± 0. 07 mg /g OH DPPH O 2-3 3 FRAP Fe 3 + O 2-90. 7% 2 g /ml OH DPPH 84. 5% 82. 2% Fe 3 + DPPH O 2 - OH 1 Valko M Leibfritz D Moncol J et al. Free radicals and antioxidants in normal physiological functions and human V C V E disease J. International Journal of Biochemistry & Cell Biology 2007 39 1 44-84.
2 期 王 华 等 : 猕 猴 桃 果 肉 提 取 物 抗 氧 化 活 性 研 究 149 2. 1 70-76. J. 2007 35 35 11454-11457. 3. J. OH J. 2005 26 2010 16 3 144-146. 4. J. 2010 31 4 187-188. 5. 2008 36 29 12755-12756. N- J. 1988 10 1 50. 6. Radical Biology & Medicine 2000 29 9 900-912. J. 2011 39 29 17773-17774. 7 Wolfe K Wu X Z Liu R H. Antioxidant activity of apple peels J. Agricultural and Food Chemistry 2003 51 3 609-614. 8 Bekhita E D. 703-717. J. 2007 28 10 61-63. 18 Whang J I Moon H I Zee O P. Phytochemical constitu- 9. ents of Aetinidia arguta J. Korean Journal of Pharma- J. 2010 10 3 372-374. 10 Halliwell B Gutteridge J M C Aruoma O I. The deoxcyribose method A simple 'test-tube' assay for determination of rate constants for reactions of hydroxyi radials J. Analytical biochemistry 1987 165 215-219. 11 Brandwilliams W Cuvelier M E Berset C. Use of a freeradical method to evaluate antioxidant activity J. Food Science and Technology-Lebensmittel-Wissenschaft and Technologie 1995 28 1 25-30. 12. O 2-52. OH J. 1993 1 25. 13 Benzie I F F Strain J J. The Ferric reducing ability of plasma FRAP as a measure of antioxidant power The FRAP assay J. Analytical Biochemistry 1996 239 14. O 2-2 121-124. 15. J. 16 Lebeau J Furman C Bemier J L et al. Aniioxidant properties of di-tert-butylhydroxylated flavonoids J. Free 17 Owen R W Haubner R Mier W et al. Isolation structure elucidation and antioxidant potential of the major phenolic and flavonoid compounds in brined olive drupes J. Food and Chemical Toxicology 2003 41 5 cognosy 2000 31 3 357-363. 19 Webby R F. A flavonol triglycoside from Actinidia arguta var. giraldii J. Phytochemisry 1991 30 7 2443-2444. 20. J. 2010 31 17 166-170. 21. J. 2011 17 5 50-22. J. 2009 30 17 98-101. 23. J. 2010 31 17 170-173.