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1 6 9 Vol. 6 No Journal of Food Safety and Quality Sep., 2015 C * 都启晶, 王艺颖, 丁君, 王成荣, 吴昊 (, ) 摘要 : 目的 C 方法 2,6- C DPPH, 结果 C, 5 cm, 100 g C mg; 3 cm, 17.28%,, ;, DPPH ;, 4 cm 结论 4 cm 关键词 : ; C; ; ; DPPH; Changes of vitamin C, isoflavone and antioxidant activity during the germination of soybean DU Qing-Jing, WANG Yi-Ying, DING Jun, WANG Cheng-Rong, WU Hao * (College of Food Science and Engineering, Qingdao Agricultural University, Qingdao , China) ABSTRACT: Objective To study the changes of vitamin C, isoflavone and antioxidant activity during the germination of soybean. Methods The optimal eating condition of soybean with a certain sprout length was determined by measuring content of vitamin C by 2,6-dichloro phenol indophenol and free radical scavenging activity by ultraviolet spectrophotometry during the period of germination of soybean sprout. Results With the increase of the length of sprout, the content of Vc was increased, reaching mg/100 g when the bud length reached 5 cm. Soy isoflavones were the highest when the bud was 3 cm, increasing by 17.28% compared with non-germinating soybean. Compared with superoxide anion radical, soybean extracts exerted the stronger scavenging activity to the DPPH radical. Two kinds of free radical scavenging ability were the largest when the bud was 4 cm. Conclusion The optimal length of soybean sprout to eat is 4 cm. KEY WORDS: soybean sprouts; vitamin C; soy isoflavone; antioxidant activity; DPPH; superoxide anion free radical 基金项目 : (SDSXDSCCYJSTX) ( ) (J14LE11) ( jch) (1207) Fund: Supported by the Modern Vegetable Industry System of Shandong Province (SDSXDSCCYJSTX), the National Natural Foundation for Young of China ( ), the Science and Technology Projects Program on High-Caliber University in Shandong Province (J14LE11), the Project of Science and Technology Program of Qingdao City ( jch) and the High-Level Personnel Scientific Research Found of Qingdao agricultural University(1207) * 通讯作者 :,, wuhaoqau@163.com *Corresponding author: WU Hao, College of Food Science and Engineering, Qingdao Agricultural University, Qingdao , China. wuhaoqau@163.com

2 9, : C 引言,,, [1],, E [2] [3], [4],,,, [5],, : ;, [6] ; γ- [7],,,, ;,,,, [8] C, 2 材料与方法 2.1 材料, 2.2 仪器与设备 AR2140 ( ); TDL-40B ( ); ( ); DK-S28 ( ); SPX-250B-Z ( ) 2.3 实验方法大豆发芽处理, 3,, 5 12 h,,, 2,,,, 2 ; 25, 12 h 40 ml 大豆发芽过程中 Vc 含量的测定 Jain Khurdiya [9], 2,6- (1) Vc, : V T Y 100 (1) W : Y Vc, mg/100 g; V 2,6-, ml; T 1 ml Vc ; W, g 大豆发芽过程中大豆异黄酮的测定 Guajardo-Flores [10],,, 260 nm A, A, C, A, 260 nm A,,, (mg)/ (g) 体外抗氧化活性的测定 (1) DPPH Mishra [11] 2 ml ( 60% ) 2 ml 0.05 mmol/l DPPH, min,, 517 nm

3 3610 6, DPPH (2), : A0 ( A2 A1) Y (%) 100 A 0 (2) : Y DPPH ; A 0 2 ml DPPH 2 ml 60% ; A 1 2 ml 2 ml ; A 2 2 ml DPPH 2 ml DPPH EC 50, EC 50 DPPH Logistic, EC 50 [12] (2) Liang [13] 4 ml 50 mmol/l Tris-HCl (ph 8.2) 1.2 ml 10 mmol/l HCl, min, 0.8 ml mmol/l,, 325 nm (60% ), 4 ml 50 mmol/l Tris-HCl (ph 8.2), 0.2 ml 10 mmol/l HCl, 1 ml, min, 0.8 ml mmol/l, 325 nm, (3), : A0 ( A1 A2) Y (%) 100 A 0 (3) : Y ; A 0 4 ml Tris-HCl 1.2 ml HCl 0.8 ml ; A 1 4 ml Tris-HCl 0.2 ml HCl 1 ml 0.8 ml ; A 2 4 ml Tris-HCl 1 ml HCl 1 ml EC 数据处理 Origin 8.0 SPSS 17.0, ANOVA, P< 结果与分析 3.1 大豆在发芽过程中 Vc 含量的变化 1, Vc, Vc, (P<0.05),, (GLDH), [14] 1 Vc Fig. 1 Change of vitamin C content during soybean germination 3.2 大豆发芽过程中大豆异黄酮含量的变化标准曲线的绘制 260 nm,, A 260, C, 2, (4) : A 260 =113.41C (4) 大豆发芽过程中异黄酮含量的变化 3,, 2 cm, 3 cm, 3 cm, (P<0.05),,

9, : C 3611 (PAL), [15], 4 Fig. 4 Free radicals scavenging ability of soybean extracts at different concentrations Fig. 2 2 Standard curve of soybean isoflavones content 3.3.2 发芽大豆 DPPH 自由基清除能力 5, DPPH, 0.

4 9, : C 3611 (PAL), [15], 4 Fig. 4 Free radicals scavenging ability of soybean extracts at different concentrations Fig. 2 2 Standard curve of soybean isoflavones content 发芽大豆 DPPH 自由基清除能力 5, DPPH, 0.08 g/ml( ),, 1 cm EC 50 = g/ml, 2 cm EC 50 = g/ml, 3 cm EC 50 = g/ml, 4 cm EC 50 = g/ml, 5 cm EC 50 = g/ml; EC 50, cm EC 50, 2 cm 4 cm, EC 50,, 2 cm 4 cm DPPH, 4 cm 3 Fig. 3 Change of soy isoflavones content during soybean germination 3.3 体外抗氧化活性大豆抗氧化 4,, ; DPPH DPPH EC 50 =0.030 g/ml, EC 50 =0.043 g/ml 5 DPPH Fig. 5 The relationship between DPPH free radical scavenging ability of bean sprouts extracts with different lengths and their concentrations

5 发芽大豆超氧阴离子自由基清除能力 (1),,,, 325 nm, 325 nm 6, (0~30 min), (30~150 min),,, 30 min,, r 2 =0.9979, 30 min,,, min -1, 30 min 7 Fig. 7 The relationship between superoxide anion free radical scavenging ability of bean sprouts extracts with different lengths and their concentrations 4 结论 6 Fig. 6 Linearity correlation between reaction time and absorbancy of reaction system (2) 7,, 0.08 g/ml( ), 1 cm EC 50 =0.051 g/ml, 2 cm EC 50 =0.094 g/ml, 3 cm EC 50 =0.053 g/ml, 4 cm EC 50 =0.018 g/ml, 5 cm EC 50 =0.046 g/ml; EC 50, 4 cm EC 50,, C, Vc ; 3 cm,, ;, DPPH,, 4 cm DPPH, 4 cm, 3 cm, 4 cm, 4 cm 参考文献 [1],,. (GABA) [J]., 2014, 17(35): Chen Z, Huang WN, Kang YF. The contents of GABA in edible beans during germination [J]. Sci Technol Food Ind, 2014, 17(35): [2] Quinhone A, Ida EI. Profile of the contents of different forms of soybean isoflavones and the effect of germination time on these compounds and the physical parameters in soybean sprouts [J]. Food Chem, 2015, 166:

9, : C 3613 [3],,,. [J]., 2011, 37(10): 62 66. Wang SY, Zhao JH, Ju XR, et al. Effect of germination on properties of soybean gel [J]. Food Ferment Ind, 2011, 37(10): 62 66.

In vivo bioavailability of essential minerals and phytase activity during soaking and germination in soybean (Glycine max L.) [J]. Aust J Crop Sci, 2014, 8(8): 1168 1174.

6 9, : C 3613 [3],,,. [J]., 2011, 37(10): Wang SY, Zhao JH, Ju XR, et al. Effect of germination on properties of soybean gel [J]. Food Ferment Ind, 2011, 37(10): [4] Saldivar X, Wang YJ, Chen P, et al. Changes in chemical composition during soybean seed development [J]. Food Chem, 2011, 124(4): [5] Kumari S, Krishnan V, Jolly M, et al. In vivo bioavailability of essential minerals and phytase activity during soaking and germination in soybean (Glycine max L.) [J]. Aust J Crop Sci, 2014, 8(8): [6] Phommalth S, Jeong YS, Kim YH, et al. Effects of light treatment on isoflavone content of germinated soybean seeds [J]. J Agric Food Chem, 2008, 56(21): [7] Zhu D, Hettiarachchy NS, Horax R, et al. Isoflavone contents in Food Chem, 2012, 134(3): [11] Mishra K, Ojha H, Chaudhury NK. Estimation of antiradical properties of antioxidants using DPPH assay: A critical review and results [J]. Food Chem, 2012, 130(4): [12] Sebaugh JL. Guidelines for accurate EC50/IC50 estimation [J]. Pharm Stat, 2011, 10(2): [13] Liang D, Zhou Q, Gong W, et al. Studies on the antioxidant and hepatoprotective activities of polysaccharides from Talinum triangulare [J]. J Ethnopharm, 2011, 136(2): [14] Xu MJ, Dong JF, Zhu MY. Effects of germination conditions on ascorbic acid level and yield of soybean sprouts [J]. J Sci Food Agric, 2005, 85(6): [15] Park NI, Xu H, Li X, et al. Overexpression of phenylalanine ammonia-lyase improves flavones production in transgenic hairy root cultures of Scutellaria baicalensis [J]. Proc Biochem, 2012, 47(12): germinated soybean seeds [J]. Plant Food Hum Nutr, 2005, 60(3): [8] Song HH, Ryu HW, Lee KJ, et al. Metabolomics investigation of flavonoid synthesis in soybean leaves depending on the growth stage [J]. Metabolomics, 2014, 10(5): [9] Jain SK, Khurdiya DS. Vitamin C enrichment of fruit juice based ready-to-serve beverages through blending of Indian gooseberry (Emblica officinalis Gaertn.) juice [J]. Plant Food Hum Nutr, 2004, 59(2): [10] Guajardo-Flores D, García-Patiño M, Serna-Guerrero D, et al. Characterization and quantification of saponins and flavonoids in sprouts, seed coats and cotyledons of germinated black beans [J]. 作者简介 ( 责任编辑 : 杨翠娜 ) 都启晶, 博士研究生, 讲师, 主要研究方向为食品安全 @qq.com 吴昊, 博士研究生, 副教授, 主要研究方向为果蔬贮藏与加工 wuhaoqau@163.com

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%