摘要
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。
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.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2015年第2期245-250,共6页
Chinese Journal of Analytical Chemistry
基金
863项目(No.2013AA092902)
浙江省自然科学基金(No.LQ13B050004)
宁波市自然科学基金(No.2014A610179)
宁波大学王宽诚幸福基金资助~~
关键词
抗氧化
亚麻籽
高效液相色谱法
2
2-二苯基-1-苦肼基-高效液相色谱法
木酚素
Antioxidant
Flaxseed
High performance liquid chromatography
2,2-Diphenyl-1-picrylhydrazyl
High performance liquid chromatography assay
Lignan
