一测多评法同时测定野菊花中12个成分

Determination of twelve components in Chrysanthemum indicum by quantitative analysis of multi-components by single marker

目的:建立HPLC一测多评法同时测定野菊花中12个成分,验证此方法在野菊花药材质量控制中的应用可行性和技术适用性。方法:采用Nucleodur C18色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,流速1.0 mL·min-1,检测波长334 nm,柱温35℃。以绿原酸为内参物,分别建立该成分与5-咖啡酰奎宁酸、咖啡酸、1,3-二咖啡酰奎宁酸、木犀草苷、4,5-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、3,4-二咖啡酰奎宁酸、蒙花苷、木犀草素、芹菜素、金合欢素的相对校正因子,再分别采用外标法和一测多评法测定30批野菊花中12个成分含量,比较2种测定方法结果的差异,验证一测多评法的准确性和可行性。结果:12个成分在测定范围内线性良好,平均回收率在97.0~102.0%之间,30批野菊花中12个成分含量测定计算值与实测值间无显著差异。30批野菊花中5-咖啡酰奎宁酸、绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、木犀草苷、4,5-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、3,4-二咖啡酰奎宁酸、蒙花苷、木犀草素、芹菜素、金合欢素含量范围分别为0.015~0.368、5.464~55.199、0.023~0.474、0.020~0.313、0.194~2.491、0.058~1.049、0.654~13.395、0.705~9.379、0.526~32.614、0.149~0.762、0.087~0.305、 0.014~1.529 mg·g-1。结论:该方法准确可靠,简便可行,节约对照品及检测成本,可以作为野菊花药材多指标成分测定质量控制方法。

Objective:To establish a quantitative analysis of multi-components by single marker(QAMS) method for the determination of 12 components in Chrysanthemum indicum,and to evaluate the feasibility of QAMS method in the quality control of Chrysanthemum indicum. Methods:HPLC separation was performed on a Nucleodur C18(4.6 mm×250 mm,5 μm) column with acetonitrile-0.1% phosphoric acid as the mobile phase. The flow rate was 1. 0 mL·min-1,the column temperature was 35℃ and the detective wavelength was 334 nm. Chlorogenic acid was used as the internal reference substances. The relative correlation factors of neochlorogenic acid,caffeic acid,1,3-dicaffeoylquinic acid,cynaroside,isochlorogenic acid C,isochlorogenic acid A,isochlorogenic acid B,linarin,luteolin,apigenin and acacetin to chlorogenic acid were calculated and established. The quantitative results obtained by QAMS were compared with those obtained by the external standard method to evaluate the feasibility of QAMS method used for multi-components determination in Chrysanthemum indicum. Results:12 components showed good linear relationship in the investigated ranges. The average recoveries ranged from 97.0% to 102.0%. No significant difference was found in the quantitative results of 12 components in 30 batches of Chrysanthemum indicum determined by the calculated and real data. Neochlorogenic acid,chlorogenic acid,caffeic acid,1,3-dicaffeoylquinic acid,cynaroside,isochlorogenic acid C,isochlorogenic acid A,isochlorogenic acid B,linarin,luteolin,apigenin,acacetin showed good linear relationship in the ranges of 0.015-0.368 mg·g-1,5.464-55.199 mg·g-1,0.023-0.474 mg·g-1,0.020-0.313 mg·g-1,0.194-2.491 mg·g-1,0.058-1.049 mg·g-1,0.654-13.395 mg·g-1,0.705-9.379 mg·g-1,0.526-32.614 mg·g-1,0.149-0.762 mg·g-1, 0.087-0.305 mg·g-1,0.014-1.529 mg·g-1,respectively. Conclusion:The method established is accurate,reliable,simple,feasible and economic,which can be used for simultaneous determination of twelve constituents in Chrysanthemum indicum.