Systematic separation and purification of 18 antioxidants from Pueraria lobata flower using HSCCC target-guided by DPPH-HPLC experiment
Systematic separation and purification of 18 antioxidants from Pueraria lobata flower using HSCCC target-guided by DPPH-HPLC experiment
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DPPH-HPLC实验引导HSCCC靶标系统分离纯化葛花中18种抗氧化剂
DOI：
10.1016/j.seppur.2012.01.041
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发表时间：
2012-03-22
影响因子：
8.6
通讯作者：
Xiong, Xiang
中科院分区：
文献类型：
--
作者：
Shi, Shuyun;Ma, Yongjian;Xiong, Xiang
采用DPPH-HPLC法和高效毛细管电泳法(HSCCC)相结合的方法，从葛花乙酸乙酯部位筛选和分离抗氧化剂。在DPPH-HPLC实验的目标指导下，HSCCC采用经典洗脱、分级洗脱、挤压洗脱和循环洗脱等洗脱方式，分离得到18种异黄酮类成分。经结晶分离后，所有抗氧化剂的纯度均在98.0%以上，其结构经UV、MS/MS和核磁共振确证。根据高效液相色谱图中的相对峰面积，发现化合物甘草素(2)、山茶素(4)、大豆苷(10)、3‘-羟基大豆苷元(13)和山茶素(14)是大白菜花中的主要抗氧化剂。其中1个为新化合物，均为首次从白花丹参花中分离得到。新化合物伊索立酮-7-O-β-D-吡喃葡萄糖醇-(1-gt；6)-β-D-吡喃葡萄糖苷(6)具有中等的抗氧化活性，IC50值为110.3微克/毫升。本研究结果表明，高效液相色谱与高效毛细管电泳法相结合是一种快速、高效、重复性好的从复杂天然产物中系统分离抗氧化剂的方法。(C)2012爱思唯尔B.V.保留所有权利。
A combinative method using DPPH-HPLC and HSCCC has been developed to screen and separate antioxidants from ethyl acetate fraction of Pueraria lobata flower. Under the target-guidance of DPPH-HPLC experiment, 18 isoflavones were isolated by HSCCC using several elution modes, such as classical elution, stepwise elution, extrusion elution and recycling elution. Following an additional dean-up step by crystallization, the purities of all the antioxidant compounds were all over 98.0% as determined by HPLC, and their structures were elucidated by UV, MS/MS and NMR analysis. Compounds glycitin (2), tectoridin (4), daidzin (10), 3'-hydroxydaidzein (13) and tectorigenin (14) were found to be the major antioxidants in P. lobata flower based on the relative peak areas in the HPLC chromatogram. Ten compounds including one new compound were first isolated from P. lobata flower. New compound, irisolidone-7-O-beta-D-glucopyranpsyl-(1 -> 6)-beta-D-glucopyranoside (6), showed moderate DPPH antioxidant activity with IC50 value of 110.3 mu g/ml. Results of the present study indicated that the combinative method using DPPH-HPLC and HSCCC was a speedy, efficient and reproductive technique to systematically isolate antioxidant compounds from complex natural products. (c) 2012 Elsevier B.V. All rights reserved.