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Bioactive Compounds and Antioxidant Activity of Mung Bean (Vigna Radiata L.), Soybean (Glycine Max L.) and Black Bean (Phaseolus

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Bioactive Compounds and Antioxidant Activity of Mung Bean (Vigna Radiata L.), Soybean (Glycine Max L.) and Black Bean (Phaseolus Food Technology and Economy, Engineering and Physical Properties Czech J. Food Sci., 34, 2016 (1): 68–78 doi: 10.17221/434/2015-CJFS Bioactive Compounds and Antioxidant Activity of Mung Bean (Vigna radiata L.), Soybean (Glycine max L.) and Black Bean (Phaseolus vulgaris L.) during the Germination Process Zhaohui XUE 1, Cen WANg 1, Lijuan ZHAI 1, Wancong YU 2, Huiru CHANG 1, Xiaohong KOU 1 and Fengjuan ZHOU 1 1School of Chemical Engineering and Technology, Tianjin University, Tianjin, P.R. China; 2Medical Plant Lab, Tianjin Research Center of Agricultural Biotechnology, Tianjin, P.R. China Abstract Xue Z., Wang C., Zhai L., Yu W., Chang H., Kou X., Zhou F. (2016): Bioactive compounds and antioxidant activity of mung bean (Vigna radiata L.), soybean (Glycine max L.) and black bean (Phaseolus vulgaris L.) during the germination process. Czech J. Food Sci., 34: 68–78. Bioactive compounds and antioxidant activity of germinated mung bean, soybean and black bean sprouts were inves- tigated to find out the effect of germination and the optimum germination time. We found that vitamin C gradually increased from zero. Compared to seeds, water-soluble protein and total flavonoid content showed a trend of sustained growth while vitamin E, total phenolic content increased at first and then decreased, the maximal value of total bioac- tive compound content was reached on the fifth day. The analysis of relative contribution revealed that total phenolics and total flavonoids made the highest (44.87–90.31%) contribution to total antioxidant activity. In conclusion, the optimum germination time for sprouts was 3–5 days when total bioactive compound content and antioxidant activi- ties both reached their peak values, which provide a sufficient theoretical basis for dietary processing and lay a solid foundation for continued research. Keywords: vitamin C; vitamin E; water-soluble protein; total flavonoids; total phenolics Mung bean (Vigna radiata L.), soybean (Glycine (Ve), flavonoids and phenolics (Guo et al. 2012). max L.), and black bean (Phaseolus vulgaris L.) are Vitamin E can protect polyunsaturated fatty acids popular food legumes consumed worldwide. How- against oxidant damage in cell membranes (Niki ever, processing is necessary before consumption 2014). Water-soluble protein (SP) is a critical factor due to non-nutritive factors (Vidal-Valverde et al. of food quality. During seed germination, storage pro- 2002). Germination is an economical and effective teins are mobilised to provide nutrients for seedling way to improve the bioactive compound content of growth (Wang et al. 2007). Phenolics are located in the sprouts (Mbithi-Mwikya et al. 2000; Vidal- the lipid-water interface of membranes, so they can Valverde et al. 2003). Sprouts have substantial scavenge free radicals inside and outside the cell. The nutritional benefits for the human body because flavonoids found in beans have been widely studied of their high concentration of nutrients which can due to their protective role against cardiovascular be used readily by the body (Randhir et al. 2004). diseases and cancer (Guajardo-Flores et al. 2013). Vitamin C (Vc) is an excellent antioxidant, espe- Over the last decade, several studies have been cially in a food system to maintain the active state focused on changes in bioactive compounds of beans, for many bioactive compounds, such as vitamin E when germination increase Vc and Ve content, water- Supported by the grants from the National Natural Science Foundation of China, Grants No. 31271979 and No. 31201245. Xiaohong Kou and Fengjuan Zhou contributed equally to this paper. 68 Czech J. Food Sci., 34, 2016 (1): 68–78 Food Technology and Economy, Engineering and Physical Properties doi: 10.17221/434/2015-CJFS soluble protein content, total phenolic content, and Determination of vitamin C. The vitamin C con- antioxidant activity (Wang et al. 2007; Fernandez- tent was determined by titration against 2,6-di- Orozco et al. 2008; Guo et al. 2012; Guajardo- chlorophenolindophenol (Thimmaiah 1999). The Flores et al. 2013; Vijaylaxmi 2013; Huang et sample (5 g) was blended with 1 g/kg oxalic acid al. 2014; Pajak et al. 2014). However, there were solution and homogenised for 3 minutes. After the no systematic researches on changes in bioactive mixture was centrifuged at 1200 g for 5 minutes, compounds and antioxidant activities of mung bean, The extract solution (10 ml) were filtered for titra- soybean and black bean during the germination pro- tion. The samples were titrated against the ascorbic cess. Besides, no scientific methods were available acid standard solution (1 mg/ml) calibrated with to evaluate the total bioactive compound content of 2,6-dichlorophenolindophenol solution until the sprouts in a comprehensive way. colour turned pink for 15 seconds. The Vc content This study was aimed at determining the effects of was expressed as mg/g fresh weight (FW). germination on the bioactive compounds and antioxi- Determination of vitamin E. The vitamin E was dant activities of mung bean, soybean and black bean in extracted according to He and Zhang (1997). The order to find the optimum conditions of germination, pulverised sample (5 g) was blended with 20 ml of understand the underlying mechanisms of increased 95% ethanol for 12 h and then added 1 ml of satu- antioxidant activity and provide theoretical guidance rated KOH for saponification (30°C, 15 min). The for further production. TOPSIS (Technique for Order solution was separated with 20 ml distilled water and Performance by Similarity to Ideal Solution) was also 20 ml diethyl ether. The content of Ve (µg/g FW) was introduced and tested to assess the total bioactive measured based on a colorimetric assay by adding compounds of different beans, provide data-based 0.5 ml/1 10-phenanthroline, 0.5 ml 1.0 mM FeCl3, foundation for manufacturing companies to optimally 0.5 ml 40 mM phosphoric acid to 4 ml of diluted select the germination period. extraction solution and the absorbance at 510 nm was measured (Huang et al. 2011). Determination of water-soluble protein content. MATERIal AND METHODS The fresh material (1 g) was homogenised with 5 ml of distilled water and subsequently centrifuged at Material and chemicals. Mung bean, soybean, and 2550 g for 10 min at 4°C; the suspension was removed black bean were supplied by the Midaocailai Company to measure the soluble protein content (SPC in mg/g (Jilin, China). Ascorbic acid (ASA), 2,6-dichloroindo- FW) of all sample extracts that was estimated colori- phenol (DIP), diethyl ether, tocopherol, phosphoric metrically with protein assay reagent by the Bradford acid, 1,10-orthopenanthroline, bovine serum albu- method (Bradford 1976). min (BSA), Coomassie brilliant blue (CBB), gallic Preparation of bean sprout extracts. The samples acid (GA), rutin, deoxyribose (DR), thiobarbituric (25 g) were extracted with 80 g/kg acetone (50 ml) acid (TBA), 2,2-diphenyl-1-picrylhydrazyl (DPPH), for 5 min (Julkunen-Tiitto 1985; Eberhardt et al. and 2,2'-azino-bis(3-ethylbenzthiazoline-6)sulfonic 2000; Liu et al. 2002; Liu & Sun 2003). The acetone acid (ABTS) were all provided by Sigma-Aldrich Co. extracts were evaporated using a rotary evaporator (St. Louis, USA). Ethylene Diamine Tetraacetic Acid at 45°C until the weight decreased to less than 10% (EDTA) was provided by Solarbio (Beijing, China). of the initial weight and then vacuum freeze dried to Seed germination. Bean seeds were cleaned and obtain crude extracts. The extracts were reconstituted soaked in water for 1 h at room temperature before to a final volume of 25 ml at the time of analysis. being placed into a semi-automatic germination ma- Extractions were performed in 3 replications for each chine (DYJ-S6365; Guangzhou Xiaoxiong Electrical individual sample. The extracts were prepared for Equipment Company, Foshan, China) with 50 beans the determination of total phenolics, total flavonoids per vessel. The germination process proceeded at and antioxidant capacity (Xu & Chang 2007). 25°C in the dark for 6 days. Sprouts were rinsed Determination of total phenolic content. The with running water every 1 hour. Germination rate amount of total phenolics (TPC) was analysed based was calculated as a germinated percentage of seeds: on a modified Folin-Ciocalteu colorimetric method (Singleton et al. 1999). It was determined from Germination rate (%) = (number of seeds germinated/ the regression equation of the calibration curve of /total number of seeds at beginning) × 100 gallic acid (concentrations ranging from 50 µg/ml 69 Food Technology and Economy, Engineering and Physical Properties Czech J. Food Sci., 34, 2016 (1): 68–78 doi: 10.17221/434/2015-CJFS to 500 µg/ml) and expressed as mg gallic acid equiva- AA (%) = {[1 – (Ai – Aj )]/A0} × 100 lents per g FW (mg GAE/g FW). Briefly, 125 µl of where: A – absorbance of the sample; A – absorbance of the standard gallic acid solution or distilled water diluted i j sample control; A – absorbance of the control extracts were mixed with 125 µl of Folin-Ciocalteu 0 reagents and 1.25 ml of 7 g sodium carbonate solu- Relative DPPH radical scavenging capacity (RDSC). tion/kg was then added. After incubation at room The RDSC was determined according to the meth- temperature for 30 min, the absorbance was meas- od of Brand-Williams et al. (1995) with slight ured at 760 nm. modifications. The sprout solution (1 ml) was mixed Determination of total flavonoid content. The with 1 ml of 0.1 mM DPPH in 95 g ethanol/kg and total flavonoid content (TFC) was determined based let stand in the dark for 20 minutes. Ethanol was on a colorimetric assay method (Zhishen et al. used instead of DPPH in the DPPH control. Distilled 1999) with slight modifications. Briefly, distilled water was used instead of the sample in the sample water diluted extract was mixed with 0.75 ml of 5 g control.
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