홍화종자에서 항산화성 계 화합물 Serotonin , N-(p-Coumaroyl)serotonin과 N-feruoylserotonin의 분리 및 정량분석

Determination and Isolation of Antioxidative Serotonin Derivatives, N-(p-Coumaroyl)serotonin and N-feruoylserotonin from Safflower Seeds

In the methanol extract from safflower seeds, two kinds of antioxidant were detected by preparative HPLC [ -Bondapak C μ 18 column (7.8×300 mm)]. Two unknown com-pounds were defined as CA and CB which had peaks at 22.1 min and 24.5 min, respectively. Antioxidant activity was measured by their scavenging ability on the stable free radical of 1,1-diphenly-2-picrylhydrazyl (DPPH). For bulk extraction of antioxidants, the methanol extract was frac-tionated with hexan, chloroform, ethyl acetate and butanol. The ethyl acetate fraction showing the highest DPPH radi-cal scavenging activity was further purified by silica gel column chromatography to CA and CB. By NMR analysis, CA and CB were identified as N-(p-Coumaroyl)serotonin and N-feruoylserotonin, respectively. The content of N-(p-Coumaroyl)serotonin and N-feruoylserotonin were analyzed by reverse phase HPLC using a -Bondapak C μ 18 column (3.9×300 mm) with linear gradient elution from 10% ace-tonitrile to 50% acetonitrile for 30min on UV detector at 300 nm. The contents of N-(p-Coumaroyl)serotonin and N-feruoylserotonin were 4.11 mg/g DW and 7.29 mg/g DW, respectively, and these two DPPH radical scavengers were detected only in the hull of seeds.