Isolation of antioxidants from Psoralea corylifolia fruits using high-speed counter-current chromatography guided by thin layer chromatography-antioxidant autographic assay
Isolation of antioxidants from Psoralea corylifolia fruits using high-speed counter-current chromatography guided by thin layer chromatography-antioxidant autographic assay
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DOI：
10.1016/j.chroma.2010.06.041
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发表时间：
2010-08-20
影响因子：
4.1
通讯作者：
Wang, Zhengtao
中科院分区：
文献类型：
--
作者：
Xiao, Guodong;Li, Guowen;Wang, Zhengtao
建立了高速逆流色谱（HSCCC）和薄层色谱（TLC）相结合的抗氧化剂自显影分析方法，用于分离补骨脂果实中的抗氧化成分。在薄层生物自显影技术的指导下，采用正己烷-乙酸乙酯-甲醇-水（1：1.1：1.3：1，v/v/v/v/v）为优化的两相溶剂体系，采用高速逆流色谱法成功地从紫堇果实中分离出了5种黄酮类化合物和3种香豆素类化合物。分离得到5.91 mg的补骨脂素、6.26 mg的异补骨脂素、3.19 mg的补骨脂素、0.92 mg的紫堇醇A。和2.43 mg巴瓦奇宁，相应的纯度为99.5、99.8、99.4、96.4和99.0%，以及三个亚级分。在通过制备型TLC进行另外的净化步骤后，从三个单独的亚级分中分离出0.4mg 8-异戊二烯基大豆苷元（纯度91.7%）、4.18mg新罂粟黄酮（纯度97.4%）和4.36mg异罂粟查尔酮（纯度96.8%）。化合物的结构经1H-1 NMR和13 C NMR确证。用电子自旋共振（ESR）法测定抗氧化活性的结果表明，雀花定是活性最高的抗氧化剂，其IC_（50）值为44.7 μ M。本文首次报道了用高速逆流色谱法同时分离紫堇中的8个化合物。(C)2010 Elsevier B. V.保留所有权利。
A combinative method using high-speed counter-current chromatography (HSCCC) and thin layer chromatography (TLC) as an antioxidant autographic assay was developed to separate antioxidant components from the fruits of Psoralea corylifolia. Under the guidance of TLC bioautography, eight compounds including five flavonoids and three coumarins were successfully separated from the fruits of P. corylifolia by HSCCC with an optimized two-phase solvent system, n-hexane-ethyl acetate-methanol-water (1:1.1:1.3:1, v/v/v/v). The separation produced 5.91 mg psoralen, 6.26 mg isopsoralen, 3.19 mg psoralidin, 0.92 mg corylifol A. and 2.43 mg bavachinin with corresponding purities of 99.5, 99.8, 99.4, 96.4, and 99.0%, as well as three sub-fractions, in a single run from 250 mg ethyl acetate fraction of P. corylifolia extract. Following an additional clean-up step by preparative TLC, 0.4 mg 8-prenyldaidzein (purity 91.7%), 4.18 mg neobavaisoflavone (purity 97.4%) and 4.36 mg isobavachalcone (purity 96.8%) were separated from the three individual sub-fractions. The structures of the isolated compounds were identified by H-1 NMR and C-13 NMR. The results of antioxidant activity estimation by electron spin resonance (ESR) method showed that psoralidin was the most active antioxidant with an IC50 value of 44.7 mu M. This is the first report on simultaneous separation of eight compounds from P. corylifolia by HSCCC. (C) 2010 Elsevier B.V. All rights reserved.