Isolation, Identification and Conversion Characteristics of Anti-inflammatory Active Saponin in Black Soybean Sprout treated by High Hydrostatic Pressure (HHP)

Abstract

Background: The use of germination and high hydrostatic pressure(HHP) processing can improve soyasaponin contents and physiological characteristics. However, no study has sought to identify the active compounds from soyasaponin extracts of black soybean treated by HHP after germination. Therefore, this study was performed to isolate and identify the anti-inflammatory active soyasaponin in black soybean sprouts treated with high hydrostatic pressure (HHP). Methods: Black soybeans were germinated for 4 days and subjected to 150 MPa for 24 h. Crude soyasaponin extracts were separated into using C18 column chromatography and the active compound was purified by semi-preparative HPLC. After isolating the active compounds, the secretion of inflammatory markers (nitric oxide, TNF-α, IL-1β and IL-6) by LPS-stimulated RAW 264.7 macrophages were measured. Result: The anti-inflammatory compounds were identified as Soyasaponin Bb and Bc. The anti-inflammatory compounds were identified as soyasaponin Bb and Bc. Soyasaponin Bc enhanced with increasing HHP treatment time in control. However, enzyme inactivation kept soyasaponin Bc content constantly regardless of HHP treatment time. Therefore, activation of enzyme is a major factor for increasing soyasaponin BC level and anti-inflammatory activity of soybean sprout according to HHP treatment.