硅胶柱色谱结合高速逆流色谱法分离纯化荷花中黄酮类化合物

采用硅胶柱色谱结合高速逆流色谱法分离纯化了荷花中3种黄酮类化合物。荷花粗提物先经过硅胶柱色谱初步分离,得到黄酮含量高的组分,再经过高速逆流色谱分离,以乙酸乙酯-乙醇-水-乙酸(4:1:5:0.025, v/v/v/v)为两相溶剂系统,上相为固定相,下相为流动相,在主机转速800 r/min、流速2.0 mL/min、检测波长254 nm条件下,从150 mg样品中一次性分离制备得到6.1 mg槲皮素-3-O-β-D-葡萄糖醛酸苷(I), 14.8 mg杨梅素-3-O-β-D-葡萄糖苷(II)和20.2 mg紫云英苷(III),经高效液相色谱检测其纯度分别为97.0%、95.4%、96.3%,并通过质谱和核磁共振氢谱、碳谱鉴定各化合物的结构。该方法简便、快速、节省溶剂,可以对荷花中的黄酮类化合物进行快速有效的分离纯化,具有较好的实用价值,为荷花资源的进一步开发应用提供了参考依据。

Separation and purification of flavones from Nelumbo nucifera Gaertn. by silica gel chromatography and high-speed counter-current chromatography

Three flavones were isolated and purified from Nelumbo nucifera Gaertn. by the combination of silica gel chromatography and high-speed counter-current chromatography (HSCCC). The crude extract of N. nucifera was separated by silica gel chromatography and the fraction containing flavones was obtained. Then, the fraction was separated by HSCCC with two phase solvent systems composed of ethyl acetate-ethanol-water-acetic acid (4: 1 : 5:0.025, v/v/v/v). The upper phase was as the stationary phase and the lower phase as the mobile phase. Under the conditions of a flow rate of 2.0 mL/min, while the apparatus rotated at 800 r/min and the detection wavelength was at 254 nm, 6.1 mg of quercetin-3-O-beta-D-glucuronide, 14.8 mg of myricetin-3-O-beta-D-glucopyranoside and 20. 2 mg of astragalin were obtained from 150 mg of the crude sample in one step. The purities determined by high performance liquid chromatography (HPLC) were 97.0%, 95.4% and 96.3%, respectively. The structures of the target compounds were identified by electrospray ionisation mass spectrometry (ESI-MS), 1H-nuclear magnetic resonance (1H-NMR) and 13C-nuclear magnetic resonance (13C-NMR). This method that has practical value not only saves solvent but also is convenient. It is effective in the separation of flavones from N. nucifera, and provides theoretical foundation for the further development and use of N. nucifera resources.