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1 5 1 Vol. 5 No Journal of Food Safety and Quality Jan., 2014 夏旭 1, 周爱梅 1*, 卢敏 1, 刘欣 1, 李启圣 2, 李少华 2 (1., ; 2., ) 摘要 : 目的, 方法, C (L-ascorbic acid),, DPPH 结果 1:90 (w:v) 50 s 3 h 320 W, 3.78% 结论,, VC 关键词 : ; ; ; ; Microwave-assisted extraction and antioxidative activity of flavonoids from Zizania XIA Xu 1, ZHOU Ai-Mei 1*, LU Min 1, LIU Xin 1, LI Qi-Sheng 2, LI Shao-Hua 2 (1. College of Food Science, South China Agricultural University, Guangzhou , China; 2. Guangzhou Pantang Food Co. Ltd., Guangzhou , China) ABSTRACT: Objective To investigate the microwave-assisted process of ethanol for the extraction of total flavonoids from Zizania, and the antioxidant activities of the resultant total flavonoids. Methods The total flavonoids in Zizania was measured with standard curve, the impact of solid to liquid ratio, microwave processing time, extraction time, and microwave power on the yield of flavonoids were examined by orthogonal experimental. The flavonoids were further studied by testing their total reducing power and their scavenging capacity on the DPPH free radical, hydroxyl free radical, and superoxide anion free radical compared with the L-ascorbic acid. Results The optimum extraction condition were at solid-liquid ratio of 1:90 (w/v), microwave treatment time of 50s, alcohol extraction time of 3 h, and microwave power of 320 W with the extraction rate of 3.78%. Conclusion The resultant total flavonoids exhibited some antioxidant activities at low concentrations and their antioxidant activities displayed a dose-response relationship with their concentrations, but lower than those of L-ascorbic acid. KEY WORDS: Zizania; flavonoids; microwave; extraction; antioxidative activities (Zizania latifolia (Griseb.) Stapf),,, [1] 基金项目 : (11BppZLhh ) Fund: Supported by Major Projects in Guangzhou City Science and Technology Plan (11BppZLhh ) * 通讯作者 :,, *Corresponding author: ZHOU Ai-Mei, Associate Professor, No.483, Wushan Road, TianheDistrict, South China Agricultural University, Guangzhou , China. zhouam@scau.edu.cn

2 1, : 253, 50,,, [2],, [3],,,,,,,,,, C (L-ascorbic acid),, 1 材料与方法黄酮标准曲线的绘制 :, 1 2, 345 nm, 510 nm, 345 nm Fig. 1 1 Scanning curve of rutin full wavelength 1.1 材料与试剂, ;, ; DPPH (2,2-Diphenyl-1-picrylhydrazyl) VC(L-ascorbic acid ) 1.2 主要仪器设备 UV-1800PC, ; WD800, ; FD-1PF, ; R-201, ; DXF-400, 1.3 实验方法 [4] 黄酮测定方法原理,, 2 Fig. 2 Scanning curve of Zizania total flavonoids full wavelength [5] : 25 mg, 100 ml, ml 25 ml, 5% NaNO 2 2 ml,, 6 min, 10% Al(NO 3 ) 3 2

3 254 5 ml, 6 min, 4% NaOH 20 ml,,, 15 min, 345 nm, 345 nm A (Y) c(x), : Y= 0.452X (R 2 = ) 样品中总黄酮含量的测定 NaNO 2 -Al(NO 3 ) 3 -NaOH, (1) : 样品总黄酮质量黄酮得率 (%) = 100 (1) 样品质量茭白总黄酮的提取及提取工艺的优化 90% 80,, 4 3, 茭白总黄酮抗氧化性能研究 [6] : 2 ml (50~200 μg/ml), 0.2 mol/l ph ml 1% 2 ml min,, 10% 2 ml, 3000 r/min 10 min, 5 ml 0.1% 1 ml, 10 min 700 nm, DPPH [7,8] : DPPH 19.7 mg, 70%, 500 ml, 70%,, 0.1 mmol/l DPPH, 4 4 ml DPPH 0.2 ml (50 ~ 200 μg/ml),, 30 min 517 nm (2) DPPH I, A A I % = A i j ( ) (2) : A 0 : 0.2 ml 4 ml DPPH ; A j : 0.2 ml 4 ml 70% ; A i : 0.2 ml 4 ml DPPH [9] : 2 ml 9 mmol/l FeSO 4 2 ml (50 ~ 200 μg/ml) 2 ml 9 mmol/l, 2 ml 8.8 mmol/l H 2 O 2,, min,, 510 nm (3) I, A A I% = A 0 i j ( ) (3) : A 0 : ; A j : H 2 O 2 ; A i : (O 2- ) [10,11] : 4.5 ml 0.05 mol/l Tris-HCl(pH 8.2), 4.2 ml, min, 0.3 ml 3 mmol/l ( 0.01 mmo/l HCL ), 0.01 mmo/l HCL 320 nm, 30 s, 15 min,, ΔA 0, 1mL 0 Table 1 表 1 微波辅助提取法的正交实验设计表 The orthogonal experimental design table of microwave assisted extraction (w) A (s) B (w/v) C (h) D : : :90 3.0

4 1, : 255 (50 ~ 200 μg/ml), ΔA (4) ΔA I % = ( 1 ) 100 (4) ΔA : ΔA 0 : ; ΔA:, 2 结果与分析 2.1 茭白总黄酮提取的单因素研究 3 4 (P<0.01) 160 ~ 480 W,, 480 W, 800 W, 10 ~ 40 s,, 40 s 40 ~ 50 s, 1:50 ~ 1:80(w:v),, 1:80, 0 1:90, ( ), 1.5 ~ 2.5 h,,, 2.5 h 2.5 ~3.5 h, 4 : 480 W 40 s 1: h 2.2 茭白总黄酮提取工艺的优化 2, : C( ) B( ) D( ) A( ), A 1 B 3 C 3 D 3, 1:90(w:v), 50 s, 3 h, 320 W, 3.77%, 3.78%, (P<0.01) 3, C, (P<0.01), B( ) D( ) (P<0.05), A ( ) (P>0.05) Fig. 3 3 The single factor experiment diagram under different level

5 256 5 表 2 L 9 (3 4 ) 正交设计实验极差分析表 Table 2 L 9 (3 4 )orthogonal design experiment range analysis table (w) A (s) B (w:v)c (h) D (%) 1 1(320) 1(30) 1(1:70) 1(2.0) (40) 2(1:80) 2(2.5) (50) 3(1:90) 3(3.0) (480) (640) k k k R Table 3 表 3 正交设计实验方差分析表 Orthogonal design experiment variance analysis table SS df MS F P A B * C ** D * : **, *, 2.3 茭白总黄酮抗氧化性能的研究总还原力的测定 4,, VC (P<0.01); (P<0.01), 200 μg/ml, VC 12.5%, DPPH 自由基的清除能力 5, VC DPPH (P<0.01), VC (P<0.01), IC μg/ml, 394 μg/ml, DPPH, VC 羟基自由基的清除能力 6,, VC,, (P<0.01),

6 1, : 257 VC (P<0.01), 200 μg/ml, VC 42.49% IC 50, VC IC μg/ml, 455 μg/ml,, VC 超氧阴离子自由基 (O 2- ) 清除能力 7, VC (P<0.01) VC 50 μg/ml,, 150 μg/ml,, VC (P<0.01),, IC μg/ml 6 VC Fig. 6 Scavenging effect of VC and total flavonoids of Zizania on OH free radical Fig 4 4 VC The reducing power of VC and total flavonoids of Zizania 7 VC Fig 7 Scavenging effect of VC and total flavonoids of Zizania on O 2- free radical 3 结论 5 VC DPPH Fig. 5 Scavenging effect of VC and total flavonoids of Zizania on DPPH free radical : 1:90(w:v), 50 s, 3 h, 320 W, 3.78% DPPH L- (VC),,,,,

258 5 参考文献 [1]. [J]., 2011, (5): 65-65. Xiang C. Zizania nutrition and diet [J]. Cook Knowledge, 2011, (5): 65 65. [2],,. [J]., 1994, (4): 14. Kong QD, Ke WD, Yang BG.

Fu GM, Wan Y, Zhou JB. Comparison of methods of determining the content of flavonoids in extracts from evcommia vlmoides oliv. Leaves [J]. J Chin Inst Food Sci Tech, 2006, 6(1): 224 229. [5],. [J]., 2007, 28(7): 31 34.

7 258 5 参考文献 [1]. [J]., 2011, (5): Xiang C. Zizania nutrition and diet [J]. Cook Knowledge, 2011, (5): [2],,. [J]., 1994, (4): 14. Kong QD, Ke WD, Yang BG. Zizania resources classification [J]. Chin Seed Ind, 1994, (4): 14. [3]. [J]., 2002, (9): 41. Xiang C. Diet of Zizania [J]. Orient Herb, 2002, (9): 41. [4],,. [J]., 2006, 6(1): Fu GM, Wan Y, Zhou JB. Comparison of methods of determining the content of flavonoids in extracts from evcommia vlmoides oliv. Leaves [J]. J Chin Inst Food Sci Tech, 2006, 6(1): [5],. [J]., 2007, 28(7): Li ZZ, Liu JH. In strawberry flavoniods extraction and antioxidative activity research [J]. Food Res Dev, 2007, 28 (7): [6],,. [J]., 2006, 27(9): Mo KJ, Liu S, Chen CS. Study on antioxidant activity of ginger flavonoid [J]. Food Sci, 2006, 27(9): [7],. DPPH [J]., 2011, 32(1): Lee XJ, Cui SY. DPPH radical scavenging mechanism of ascorbic acid [J]. Food Sci, 2011, 32(1): [8] Balasundram N, Ai TY, Sambanthamurthi R, et al. Antioxidant properties of palm fruit extracts [J]. Asia Pac J Clin Nutr, 2005, 14(4): [9] Huang D, Ou B, Prior RL. The chemistry behind antioxidant capacity assays [J]. J Agr Food Chem, 2005, 53(6): [10] Tang F, Wang J, Yao X, et al. Detection of antioxidative capacity of bamboo leaf extract by scavenging hydroxyl free radical [J]. Spectrosc Spectr Anal, 2010, 30(2): [11],,,. [J]., 2011, 36(1): Li PJ, Liu XG, Long HR, et al. Ultrasonic -assisted extraction and antioxidant activity of flavonoids from water chestnut shell [J]. Food Tech, 2011, 36(1): 作者简介 ( 责任编辑 : 张宏梁 ) 夏旭, 研究生, 主要研究方向为食品化学与营养 hnnyxiaxu@gmail.com 周爱梅, 副教授, 博士, 主要研究方向为化学与营养及水产品加工研究 zhouam@scau.edu.cn

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g 16 6 2018 11 Chinese Journal of Bioprocess Engineering Vol. 16 No. 6 Nov. 2018 doi 10. 3969 /j. issn. 1672-3678. 2018. 06. 004 1 2 1 1 1. 330200 2. 330006 4 Box-Behnken 1 25 g /ml 121 92 min 2 6. 284%