制备型高效液相色谱法分离纯化绿原酸

Separation and purification of chlorogenic acid by preparative high performance liquid chromatography

采用制备高效液相色谱技术,从杜仲叶提取物中分离纯化得到高纯活性成分绿原酸。对制备色谱的洗脱方式、洗脱剂组成、浓度、洗脱速度等参数进行优化,提出并应用了流动相速度梯度洗脱方法。最佳操作条件为:采用流速梯度进行洗脱,流速先为28mL/min,然后为45mL/min;流动相为15%的有机酸B和0.1%的有机酸E(体积分数);测定波长为254nm;进样体积为10mL;进样量为2500mg。通过该工艺分离纯化,制备回收率为83.3%,相对标准偏差为3.1%,绿原酸的纯度达98.61%,同时可得到杜仲叶黄酮、桃叶珊瑚甙等副产品。

Preparative high performance liquid chromatography (PHPLC) technique was used to separate and purify high purity of chlorogenic acid from Eucommia ulmoides Oliv. The elution mode, composition of the mobile phase, content of the organic acid and the flow-rate of the mobile phase were optimized and thus a new flow rate gradient was proposed. The optimum operation parameters were: the flow rate gradient of 28 mL/min first and 45 mL/min later, the mobile phase of 15% alcohol B added with 0.1% organic acid E(volume fraction), the determination wavelength of 254 nm, the injecting volume of 10 mL and the injecting mass of 2 500 mg. The average recovery rate was 83.8% and relative standard deviation(RSD) was 3.1%. High purity product of chlorogenic acid ((98.61%)) was obtained and some by-products such as aucubin and flavone were also obtained.