高速逆流色谱分离纯化荷叶黄酮槲皮素

Isolation and preparation of quercetin from flavoids of lotus leaf by high speed counter-current chromatography

以含量为4.4%的荷叶黄酮粗粉为原料,在优化高效液相色谱分析荷叶黄酮槲皮素条件的基础上,通过对不同溶剂体系的优选及操作参数的优化,建立了高速逆流色谱分离纯化荷叶黄酮槲皮素的技术方法。结果表明,采用流动相甲醇和0.4%磷酸、梯度(0min90%甲醇→10min60%甲醇→20min40%甲醇→40min20%甲醇)洗脱、检测波长360nm、流速1.0mL/min、柱温30℃、进样量20μL的技术参数可作为高效液相色谱分析荷叶黄酮槲皮素的方法;采用氯仿:甲醇:水(体积比8:10:5)为溶剂系统(上相为固定相,下相为流动相)、流速2mL/min、转速850r/min、进样量150mg的高速逆流色谱操作技术参数,可分离制备荷叶黄酮槲皮素单体,其纯度大于99%。

A high speed counter-current chromatography（HSCCC） method for preparative isolation and purification of quercetin were established,by a method of varieties of solute systems improved and technology parameters optimized,based on high performance liquid chromatography（HPLC） method which was applied for analysizing quercetin of flavonoid from lotus.For this project,quercetin was isolated from raw extracts of lotus leaves containing 4.4% of flavonoids.Mobile phase：methanol and 0.4% phosphoric acid,elute with gradient（0min 90%methanol→10min 60% methanol→20min 40%methanol→40min 20%methanol）,detection wavelength：360nm,flow rate：1.0mL/min,column temperature：30℃,injection volume：20μL,as the optimized HPLC conditions.Chloroform-methanol-water 8：10：5（v）,used as a two-phase solvent system for high speed counter-current chromatography（HSCCC）.The upper phase was used as the stationary phase,while the lower phase as the mobile phase.The separation processes were performed at a flow-rate of 2mL/min and a revolution speed of 850r/min with injection volume of 150mg.The purity of 99% was yielded.