Systematic separation and purification of 18 antioxidants from Pueraria lobata flower using HSCCC target-guided by DPPH-HPLC experiment

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• 作者：Shuyun Shi^a ; ^b ; ^{shuyshi@gmail.com} ; Yongjian Ma^b ; Yuping Zhang^b ; Liangliang Liu^b ; Qi Liu^b ; Mijun Peng^c ; ^{pengmj163@163.com} ; Xiang Xiong^a
• 关键词：Pueraria lobata flower ; DPPH ; HPLC ; HSCCC ; Isoflavone ; Antioxidant
• 刊名：Separation and Purification Technology
• 出版年：2012
• 期刊代码：78_13835866
• 类别：ce
• 出版时间：22 March, 2012
• 卷：89
• 期：Complete
• 页码：225-233
• 文件大小：491 K

摘要

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 clean-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鈥?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-尾-d-glucopyranpsyl-(1鈫?)-尾-d-glucopyranoside (6), showed moderate DPPH antioxidant activity with IC₅₀ value of 110.3 渭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.