高效制备液相色谱法从荷叶中分离制备黄酮类化合物

采用高效制备液相色谱法从荷叶(Nelumbo nucifera Gaertn)中分离制备荷叶黄酮类化合物。用60%乙醇回流提取荷叶，粗提液浓缩后经D-101柱及聚酰胺柱色谱分离，再在Symmetry PrepTM C18柱上分离,以水-乙腈为流动相进行梯度洗脱（流速为5.0 mL/min）,得到了3种黄酮类化合物。经紫外光谱、红外光谱、核磁共振及质谱分析,确定该3种物质分别为金丝桃苷、异槲皮苷和紫云英苷。所制备的3种化合物的纯度都在97%以上,其中紫云英苷为首次从荷叶中分离得到。

Isolation and Preparation of Flavonoids from the Leaves of Nelumbo nucifera Gaertn by Preparative Reversed- Phase High Performance Liquid Chromatography

It has been confirmed that flavonoids in the leaves of Nelumbo nucifera Gaertn(lotus leaves) have many pharmacological activities.Currently,total flavones in the leaves of Nelumbo nucifera Gaertn have been studied extensively,however,only a few researches were(able) to investigate the individual components in it.At first,crude extract was obtained from lotus leaves by reflux extraction using 60% ethanol for three times.Then,the concentrated crude extract was separated on a D-101 column(eluted with 70% ethanol) and a polyamide column(step gradient 15% to 90% ethanol).The Fr-1 fraction was obtained from the eluate of 45% ethanol and was subjected to a preparative reversed-phase high performance liquid chromatograph(RP-HPLC) for the isolation of target components.The preparation of the individual flavonoids was carried out on an RP-HPLC with a Symmetry Prep~(TM) C_(18) column,and the mobile phase was wateracetonitrile at a flow rate of 5.0 mL/min.Three compounds were identified with ultra violet absorbance(UV),infrared(IR),nuclear magnetic resonance(NMR) and mass spectrometry(MS).They were hyperin,isoquercetin and astragalin.To our knowledge that astragalin was the first time successfully isolated from this plant.The purity of the three compounds was all over 97%.