The extraction conditions were as follows:solvent system:petroleum ether-ethyl acetatemethanol-water (2∶ 8∶ 2∶ 8,2∶ 8∶ 4∶ 6;v/v),rotation speed:800 r / min,flow speed:5.0 mL/min,detection wavelength:254 nm.The obtained fractions were identified by electrospray ionization mass spectrometry (ESI-MS),1H-nuclear magnetic resonance(NMR) and 13C-NMR.Two main compounds,3,5-dicaffeoyl quinic acid (11.2 mg) and okanin (10.7 mg) were separated from 200 mg crude extract,whose purities were 92.82％ and 97.55％ and analyzed by HPLC.And 3,5-dicaffeoyl quinic acid was separated from Coreopsis tinctoria firstly.Experimental results indicate that the method is rapid and highly efficient.%采用梯度逆流色谱技术快速分离纯化雪菊中的3,5-二咖啡酰奎宁酸和奥卡宁,以石油醚-乙酸乙酯-甲醇-水(2∶8∶2∶8,2∶8∶4∶6;v/v)为溶剂系统,上相为固定相,下相为流动相,转速800 r/min,流速5.0mL/min,检测波长254 nm.所得馏分通过电喷雾电离(ESI)质谱和核磁共振谱(NMR)鉴定化合物的结构.从200mg粗提物中一次分离得到两个主要化合物:3,5-二咖啡酰奎宁酸(11.2 mg)和奥卡宁(10.7 mg),且3,5-二咖啡酰奎宁酸为首次从雪菊中分离得到;经高效液相色谱法分析,其纯度分别为92.82％和97.55％.研究结果表明,应用梯度逆流色谱分离雪菊中的生物活性成分快速高效.
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