CAJ | 학술논문

为了更加迅速地筛选到亚麻籽复杂提取物中的抗氧化活性物质及其活性强弱,以开环异落松树脂酚二葡萄糖苷(SDG)、开环异落叶松树脂酚(SECO)和肠二醇(ED)3种木酚素为研究对象,建立了抗氧化活性能力筛选的2,2-二苯基-1-苦肼基-高效液相色谱法( DPPH-HPLC法)。液相色谱条件为：流动相为乙腈和水,经反相色谱柱Waters XBridge C18分离,检测波长为280 nm。 DPPH-HPLC法的步骤是：将单一或者混合抗氧化剂分别与DPPH混合,放置20 min,作为待测液,将同样量的相应抗氧化剂作为对照液,分别进行液相色谱分析获得其含量,计算出相应的抗氧化剂清除率( SRA),活性顺序为：SDG>SECO>ED。结合超高效液相色谱-飞行时间质谱联用法,鉴定出亚麻籽中5种抗氧化物,筛选其抗氧化能力分别为：SDG 异构体(5)>SDG (4)>7α-[(β-D-glupyranosyl) oxy]-1-methoxyisolariciresinol (1)>(6R,7R,8S)-1-Methoxyisolariciresinol (2)>蜀葵苷元二葡萄糖苷(3)。结果表明,建立的DPPH-HPLC法能够有效完成复杂亚麻籽提取物的抗氧化活性物质筛选。
위료경가신속지사선도아마자복잡제취물중적항양화활성물질급기활성강약,이개배이락송수지분이포도당감(SDG)、개배이락협송수지분(SECO)화장이순(ED)3충목분소위연구대상,건립료항양화활성능력사선적2,2-이분기-1-고정기-고효액상색보법( DPPH-HPLC법)。액상색보조건위：류동상위을정화수,경반상색보주Waters XBridge C18분리,검측파장위280 nm。 DPPH-HPLC법적보취시：장단일혹자혼합항양화제분별여DPPH혼합,방치20 min,작위대측액,장동양량적상응항양화제작위대조액,분별진행액상색보분석획득기함량,계산출상응적항양화제청제솔( SRA),활성순서위：SDG>SECO>ED。결합초고효액상색보-비행시간질보련용법,감정출아마자중5충항양화물,사선기항양화능력분별위：SDG 이구체(5)>SDG (4)>7α-[(β-D-glupyranosyl) oxy]-1-methoxyisolariciresinol (1)>(6R,7R,8S)-1-Methoxyisolariciresinol (2)>촉규감원이포도당감(3)。결과표명,건립적DPPH-HPLC법능구유효완성복잡아마자제취물적항양화활성물질사선。
The free radical scavenging effect of flaxseed was screened by HPLC-DPPH ( 2 , 2-diphenyl-1-picrylhydrazyl-high performance liquid chromatography assay ) and colorimetric DPPH methods. To test the effectiveness of the approach, three Lignans ( secoisolariciresinol diglucoside ( SDG ) , secoisolariciresinol ( SECO) and enterodiol( ED) ) with antioxidative properties were investigated both in monomer and mixture. HPLC conditions were optimized using following methods: Waters XBridge C18 was used as stationary phase, acetonil/H2 O was used as mobile phase and detective wavelength was set at 280 nm. Antioxidant activity of standards was investigated by reaction with or without DPPH radical for 20 min as sample and control, respectively. Both of them were analyzed by high performance liquid chromatography. According to the changes of amount of sample and control, the antioxidant activities of standards were calculated as following order:SDG>SECO>ED. Based on above DPPH-HPLC assay and UPLC-Q-TOF-MS, antioxidants extracted from flaxseed were separated, identified and screened. The radical scavenging activities were in the following order:SDG isomer (5)>SDG (4)>7α-[(β-D-glupyranosyl) oxy]-1-methoxyisolariciresinol (1)>(6R,7R, 8S)-1-methoxyisolariciresinol (2)>herbacetindiglucoside (3). It indicated that the HPLC-DPPH assay could be successfully used for the antioxidant activity screening of complex flaxseed extract.