摘要
目的:建立高效逆流色谱法(HPCCC)从白益母草中分离制备4个抗氧化成分。方法:白益母草95%乙醇水提取物经D101大孔树脂柱色谱,先用水和45%乙醇水各1000 mL洗脱,然后用70%乙醇水(5000 mL)洗脱。70%乙醇水洗脱液减压回收至干,用HPCCC法以正己烷-正丁醇-水-冰醋酸(1∶2∶1∶0.1)为两相溶剂系统,在主机转速1000 r·min^(–1),检测波长261 nm条件下进行分离制备,得到化合物1~4,经谱学数据分析,确定其结构式。同时,利用1,1-二苯基-2-三硝基苯肼(DPPH)法测定化合物1~4的抗氧化活性。结果:从白益母草70%乙醇水洗脱液中分离得到4个化合物,分别为5,7,4′-三羟基黄酮-7-O-(6′′-O-反式-咖啡酰基)-β-葡萄糖苷(1)、5,7,4′-三羟基黄酮-7-O-(2′′,6′′-O-反式-二香豆酰基)-β-葡萄糖苷(2)、5,7,4′-三羟基黄酮-7-O-(6′′-O-反式-香豆酰基)-葡萄糖苷(3)和毛蕊花苷(4)。化合物1~4表现出显著的抗氧化活性,其有效半数浓度(EC_(50))平均值分别为(22.9±2.7)、(26.2±3.4)、(24.4±3.1)和(19.5±1.8)μg·mL^(-1)。结论:建立的方法可高效分离纯化白益母草中可作为天然抗氧化剂的4个化合物。
Objective:To develop an high performance counter current chromatography(HPCCC) method combined with macroporous resin chromatography for separation and preparation of antioxidant compounds from Panzeria alashanica. Methods:The 95% ethanol extract of Panzeria alashanica was subjected to D101 macroporous resin column and was eluted with water(1 000 mL),45% ethanol(1 000 mL) and 70% ethanol(5 000 mL). The 70% ethanol eluate was recovered to dry under reduced pressure,which was isolated by HPCCC with n-hexane-n-butanol–water–glacial acetic acid(1∶2∶1∶0.1) as a two-phase solvent system with a 1 000 r·min^(-1) of rotation rate. The antioxidant activities of compounds 1-4 isolated from the 70% ethanol fraction of Panzeria alashanica were tested using 1,1-diphenyl-2-picrylhydrazyl(DPPH). Results:Four compounds 1-4,namely 5,7,4′-trihydroxyflavone-7-O-(6′′-O-[E]-caffeyl)-β-glucopyranoside(1),5,7,4′-trihydroxyflavone-7-O-(2′′,6′′-O-[E]-dicoumaroyl)-β-glucopyranoside(2),5,7,4′-trihydroxyflavone-7-O-(6′′-O-[E]-coumaroyl)-β-glucopyranoside(3) and verbascoside(4),were isolated from the 70% ethanol fraction of Panzeria alashanica. Compounds 1-4 demonstrated significant antioxidant activity. The mean±SD values of EC_(50) were(22.9±2.7),(26.2±3.4),(24.4±3.1)and(19.5±1.8)μg·mL^(-1) for compounds 1-4,respectively. Conclusion: The established method can effectively separate and purify four compounds which can be used as natural antioxidants.
作者
王青虎
陈敖日格勒
张国军
包娜布其
胡毕斯哈拉图
WANG Qing-hu;CHEN Ao-ri-ge-le;ZHANG Guo-jun;BAO Na-bu-qi;Hu-bi-si-ha-la-tu(College of Traditional Mongolian Medicine,Inner Mongolia University for Nationalities,Tongliao 028000,China;Baotou Mongolian Traditional Chinese medicine hospital,Baotou 014000,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2021年第9期1583-1589,共7页
Chinese Journal of Pharmaceutical Analysis
基金
内蒙古自治区自然科学基金项目(2018MS08034)
内蒙古自治区卫生科技项目(201701105)。
