Optimal Extraction Condition of Anthocyanins in Soybean (Glycine max) with Black Seed Coats

Soybeans (Glycine max) with a black seed coat have been widely utilized as food source and as a medicinal herbs in Korea. The pigmentation in the seed coat of black soybean is due to accumulate anthocyanins in the epidermis palisade layer. The anthocyanin content and composition of the black soybean seed coat are considered as a standard. of quality evaluation of black soybean. The main objective of this study was to investigate the optimal condition for an extraction method of anthocyanins and compare anthocyanin quantity and composition within black soybean varieties and germplasms. In the test of extraction solvent, absorbance at 530 nm and Hunter's a value were increased as increasing the concentration of MeOH, but Hunter's Land b values were the exact opposite of absorbance and Hunter's a values. There was no significant difference for anthocyanin contents from 1% HCl - to 1% HCl - 80% MeOH. In the aspects of anthocyanin contents and HPLC peak resolution, 1% HCl - 20% MeOH extraction solution was the most suitable solvent. Among the 5 kinds of extraction method using 1% HCl - 20% MeOH solution, the anthocyanin contents of room temperature extraction at 72 h was the highest among the methods. High extraction temperature, sonication and reflux method influenced on the decrease of anthocyanin contents because of breakdown of anthocyanins. There was no significant difference for extraction time between 12 h and 24 h. However, the optimal extraction condition were at room temperature for 12 h. The anthocyanin contents in seed coats of black soybean were determined on the basis of HPLC peak area at 530 nm. Ten black soybean varieties and germplasms were tested with optimal conditions founded in this study. On the basis of antocyanin component, these can be classified into three groups; C3G, C3G + D3G and C3G + D3G + Pt3G. The total anthocyanin content in seed coats ranged from 1.58 to 10.62 mg/g of seed. The total anthocyanin content of the variety "Geomjeongol" was about 7 times higher than that of variety "Heugchong". Information for extraction method and diversity in antocyanin of soybean seed coats can be used for future research for germplasm evaluation and development of high quality black soybean varieties.