工业色谱法纯化山银花中绿原酸的工艺研究

Study of the New Process of the Purification by Industrial Chromatography of Chlorogenic Acid in Lonicera Macranthoides

目的探索工业色谱法纯化山银花中绿原酸的最优工艺参数。方法以绿原酸的转移率为评价指标,以山银花水提液过大孔树脂后的20%乙醇洗脱液为原料液,采用单因素及响应面设计试验,对采用工业色谱纯化山银花中绿原酸工艺中的原料液浓缩程度、固定相类型、流动相乙醇浓度、乙醇流速、进样量等主要工艺参数进行优化,确定了最优工业色谱纯化山银花中绿原酸的工艺条件。结果工业色谱纯化山银花中绿原酸的最优工艺条件:以60℃,-0.1 MPa为浓缩条件,将原料液浓缩至原体积的0.166 7(1/6),工业色谱UV检测波长为230 nm,以PIPO-02色谱填料为固定相,以18.54%乙醇作为流动相,流速为14.57 m L·min^(-1),进样量为13.15 mg。该工艺绿原酸的平均转移率为95.49%,RSD=1.91%,绿原酸产品纯度为97.45%,RSD=3.30%。结论经过工艺优化与验证,创立了一条工业色谱纯化山银花中绿原酸工艺路线,该工艺制备的绿原酸产品纯度>97%,绿原酸转移率高,工艺简单、无毒,溶剂易于回收,适合于山银花中绿原酸纯化的工业化生产。

OBJECTIVE To explore the optimal process of the purification by industrial chromatography of chlorogenic acid in Lonicera macranthoides.METHODS The transport rate of chlorogenic acid was used as main assessment index,the eluent of 20%ethanol solution of macroporous resin,which first gained from the water extract solution of Lonicera macranthoides,was used as raw material,single factor and response surface test was adopt to design this test,these main factors,which have effect on the process of the purification by industrial chromatography of chlorogenic acid in Lonicera macranthoides,such as the type of stationary phase,the concentration of ethanol in mobile phase,the flow rate of mobile phase,the sample amount,were optimized.The optimum condition of the process of the purification by industrial chromatography of chlorogenic acid in Lonicera macranthoides was determined.RESULTS The optimum conditions of the process of the purification by industrial chromatography of chlorogenic acid in Lonicera macranthoides were as follows：the condense condition was 60℃,-0.1 MPa,the volume of raw material should be concentrated to one-sixth of its original volume,the UV determine wavelength of industrial chromatography was 230 nm,the chromatography packing of PIPO-02 was used as the stationary phase,18.54%ethanol was used as the mobile phase,the flow rate of mobile phase was 14.5 m L·min^-1,the sample amount was 13.15 mg.In this process the average data-transfer rate of chlorogenic acid was 95.49%（RSD=1.91%）,the purity of product was 97.45%（RSD=3.30%）.CONCLUSION A new route of the process of the purification by industrial chromatography of chlorogenic acid from Lonicera macranthoides has been established through optimization and verification experiment.The process,with a high transport rate of chlorogenic acid,is simple,healthy,the solvent is easy to recycle,and the process is suitable for industrialize production of the purification by industrial chromatography of chlorogenic acid in Lonicera macranthoides.