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高速逆流色谱法分离紫锥菊花色苷及其抗氧化性研究
引用本文:李佳银,罗晋,李觅路,陆英,刘仲华.高速逆流色谱法分离紫锥菊花色苷及其抗氧化性研究[J].分析测试学报,2012,31(1):45-50.
作者姓名:李佳银  罗晋  李觅路  陆英  刘仲华
作者单位:1.湖南农业大学国家植物功能成分利用工程技术研究中心; 2.湖南农业大学园艺园林学院
基金项目:湖南省重大科技专项资助(NK07006)
摘    要:建立了高速逆流色谱(HSCCC)分离制备紫锥菊花色苷类化合物的方法,并对所获得的2个花色苷单体进行了体外抗氧化性实验。以新鲜紫锥菊花瓣为原料,含0.1%HCl的60%乙醇为溶剂避光冷浸提取,经乙酸乙酯萃取和D101大孔吸附树脂(100 mL,2 cm×30 cm)纯化后,得2.1 g紫锥菊花色苷提取物干粉样品。以水-正丁醇-甲基叔丁基甲醚-乙腈-三氟乙酸(6∶3∶2∶1∶0.001)为HSCCC分离溶剂系统,上相为固定相,下相为流动相,流速2.0 mL/min,进样量160 mg,通过一次分离得到2种花色苷单体化合物,经HPLC检测其纯度分别达95.1%(9.8 mg)、98.2%(14.3 mg),MS及NMR技术鉴定其结构分别为矢车菊素-3-O-β-D葡萄糖苷(化合物1)和矢车菊素-3-O-(6″-O-丙二酰-β-D葡萄糖苷)(化合物2)。以Vc为对照组,对所获得的2种花色苷单体化合物进行了1,1-二苯基-2苦肼基(DPPH.)体外抗氧化性能评价,结果显示2种花色苷对DPPH.的半清除率(EC50)均小于10 mg/L,小于对照样Vc,表明2种花色苷均具有较强的自由基清除作用,且化合物1的清除能力强于化合物2。

关 键 词:高速逆流色谱  紫锥菊  花色苷  DPPH.  抗氧化性

Separation of Anthocyanins in Echinacea Purpurea Flower by High Speed Counter current Chromatography and the Research on the Antioxidant Activity of the Anthocyanins
LI Jia-yin,LUO Jin,LI Mi-lu,LU Ying,LIU Zhong-hua.Separation of Anthocyanins in Echinacea Purpurea Flower by High Speed Counter current Chromatography and the Research on the Antioxidant Activity of the Anthocyanins[J].Journal of Instrumental Analysis,2012,31(1):45-50.
Authors:LI Jia-yin  LUO Jin  LI Mi-lu  LU Ying  LIU Zhong-hua
Institution:1,2 (1.National Research Center of Engineering Technology for Utilization of Functional Ingredients from Botanicals,Hunan Agricultural University,Changsha 410128,China;2.College of Horticulture and Landscape,Hunan Agricultural University,Changsha 410128,China)
Abstract:An efficient method for the isolation and preparation of two anthocyanins,cyaniding 3-O-(β-D-giucopyranoside)(compound 1) and cyaniding 3-O-(6″-O-malonyl-β-D-glucopyranoside)(compound 2),from Echinacea purpurea flower by high speed counter-current chromatography(HSCCC) was established.The fresh flower was extracted with 60% ethanol(containing 0.1% HCl) at low temperature and dark conditions.2.1 g of crude sample for HSCCC was prepared after extracted with ethyl acetate and passed through an adsorbing column(2 cm× 30 cm,containing 100 mL D101 macroporous resin).Then water-n-butanol-methyl tert-butyl ether-acetonitrile-trifluoroacetic acid(6 ∶3 ∶2 ∶1 ∶0.001) was used as the two-phase solvent system,in which the upper phase was used as the stationary phase,while the lower phase was used as the mobile phase,the flow rate was set at 2.0 mL/min,and the revolution speed was 850 r/min.9.8 mg compound 1 with purity of 95.1% and 14.3 mg compound 2 with purity of 98.2% were obtained from 160 mg crude sample by one-step,as determined by high-performance liquid chromatography(HPLC).The chemical structures were identified by mass spectrometry(MS) and nuclear magnetic resonance imaging(NMR).Additionally,the antioxidant activities of the two compounds were also evaluated by the method of 1,1-diphenyl-2-picrylhydrazyl(DPPH·) radical scavenging assay with Vc as the control group.The results of antioxidative activity test showed that the two compounds have strong capacity of scavenging DPPH(EC50<10 mg/L).Both of their EC50 were lower than that of Vc,and the scavenging ability of compound 1 was stronger than that of compound 2.
Keywords:high speed counter-current chromatography  Echinacea purpurea  anthocyanins  DPPH·  antioxidant activity
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