聚酰胺色谱结合高速逆流色谱分离制备萹蓄黄酮类化合物

Isolation and Purification of Flavones from Polygonum aviculare L. by Polyamide Chromatography Combined with High-Speed Counter-Current Chromatography

采用聚酰胺色谱结合高速逆流色谱法分离纯化了萹蓄中3种黄酮类化合物,建立了快速分离制备萹蓄中3种黄酮类化合物的方法。通过聚酰胺柱色谱富集黄酮类成分,再经过高速逆流色谱分离,以乙酸乙酯-甲醇-水-甲酸(体积比为4∶1∶5∶0.1)组成的二相系统作为固定相与流动相,在主机转速为850 rpm,流速为2.0m L/min,检测波长为254 nm的条件下制备样品。从150 mg富集黄酮成分的馏分中,一次性分离制备得到纯度为94.86%的杨梅树皮苷(myricitrin)7.5 mg,94.28%的黄芪苷(astragalin)13.8 mg,91.86%的合欢草素1(desmanthin-1)20.6 mg。所得馏分经高效液相色谱法(HPLC)检测纯度,并经MS和NMR鉴定化合物的结构。该方法简便、快速,所得产物纯度高,适合于黄酮类化合物的制备分离。

In this study,three flavones were isolated and purified from Polygonum aviculare L. by polyamide chromatography combined with high-speed counter-current chromatography（ HSCCC）. A rapid and efficient method was established for preparative isolation and purification of three flavonoids from P. aviculare. The crude extract of P. aviculare was objected to polyamide chromatography to afford the flavones-enriching fraction. After that,the fraction was further separated by HSCCC with two phase solvent systems composed of ethyl acetate-methyl alcohol- water-formic acid（ 4∶ 1∶ 5∶ 0. 1,v / v）.The lower phase as the mobile phase was operated at a flow rate of 2. 0 m L / min,while the apparatus rotated at 850 rpm and the detection wavelength was 254 nm. As a result,7. 5 mg of myricitrin,13. 8 mg of astragalin,20. 6 mg of desmanthin-1 were purified from 150 mg of the flavones-enriching fraction and each with 94. 86%,94. 28% and 91. 86% purity as determined by HPLC. These compounds were identified by mass spectrometry（ MS） and1H-nuclear magnetic resonance（1H NMR）. The method was simple,rapid and suitable for the preparative separation of flavonoids.