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Foods 2018, 7, 174 4 of 10 standards of soybean isoflavones (Sigma Aldrich Co., St. Louis, MO, USA). All samples were analyzed in triplicate and results were expressed as milligrams per gram (mg/g). 2.6. DPPH Free Radical Scavenging Capacity The antioxidant capacity was determined on the basis of the scavenging capacity of the stable 2, 2-diphenyl-1 picryl hydrazyl (DPPH) free radical according to methods described by Braca et al. [20] with slight modifications. One mL of stock solution was added to 3 mL of DPPH. The mixture was shaken vigorously and left to stand at room temperature in the dark for 30 min. The absorbance was measured at 517 nm using a spectrophotometer (UV-1800 240 V, Shimadzu Corporation, Kyoto, Japan). The percent inhibition activity were calculated against a blank sample using the following equation: inhibition (%) = (blank sample-extract sample/blank sample) × 100. 2.7. Ferric Reduction Antioxidative Power (FRAP Assay) The FRAP was determined according to methods described by Benzie and Strain [21]. The FRAP reagent contained 20 mL of a 10 mM TPTZ (2,4,6-tripyridyl-s-triazine) solution in 40 mM HCl, 20 mL of 20 mM iron (III) chloride hexahydrate and 200 mL of 0.3 M acetate buffer at pH 3.6. The FRAP reagent was incubated at 37 ◦C in a water bath. Aliquots from stock solution of 5 mL were mixed with 45 μL of FRAP reagent. The absorbance was measured at 593 nm using spectrophotometer. Ferrous sulfate heptahydrate was used as a standard for the calibration curve and the results were expressed as FRAP value (μM Fe2+). 2.8. Statistical Analysis Data are reported as mean ± standard deviation from triplicate analysis. Analysis of variance (ANOVA) accompanied with LSD and Tukey tests (SPSS, Version 15, IBM, New York, NY, USA)) were conducted to identify the significant differences among the samples (p < 0.05). 3. Results and Discussion 3.1. Effect of Artificial LED Light on the Accumulation of Total Phenol, Isoflavones Content and Antioxidant Capacity Light is the important environmental que to improve the bioactive compounds in plant materials [22]. Light stimulate the enzyme activation and regulate the enzyme synthesize pathways, such as PAL (phenylalanine ammonia-lyase) activity in phenyl-propanoid pathways, which promote the bioactive compound accumulation in plant [23]. The isoflvones and total phenol content of the soybean sprouts grown under artificial LED light were shown in Table 1 and Figure 2. It is clearly depicted that isoflavones and total phenol were accumulated higher in the soybean sprout grown under blue LED light compared to the green and florescent light. The isoflavones, such as daidzin, glycitin, genistin, daidzein, glycerin and genistein, were significantly (p < 0.05) increased at the five and six DAS. A reduction of the isoflavones contents were observed with increasing DAS. Studies showed that blue light is the most effective lighting source to synthesis flavonoid compounds by stimulating PAL, CHS (chalcone synthesis) and DFR (dihydroflavonol-4-reductage) gene expression [24]. Chi et al. [25], showed that soybean sprouts grown under artificial LED light condition contained higher isoflavones content than those grown under florescent light. Cevallos and Cisneros [26] found higher phenolic contentofsoybeansproutatsevendaysofgermination.Additionally,Troszyn ́skaetal.[27]reported the highest concentration of polyphenols in lentils at seven days of germination. Conversely, phenolic content in mung bean decreased with germination days [28].PDF Image | Effect of Artificial LED Light and Far Infrared Irradiation on soybean
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