金银花中异绿原酸A、B、C含量的HPLC测定方法研究

The Determination of Isochlorogenic Acid A,B,and C in Lonicera japonica by HPLC

采用高效液相色谱法(HPLC)同时测定金银花中异绿原酸A、B、C的含量,并对色谱条件进行优化。结果表明,最佳色谱条件为Eclipse plus C18色谱柱(4.6mm×150mm,5μm),流动相为0.1%磷酸溶液(A)-乙腈(B),梯度洗脱,洗脱条件为0-20min,15%~30%B,20-30min,30%~50%B,流速1.0mL/min,检测波长325nm,柱温30℃,进样量10μL。在最佳的色谱条件下,实现了金银花对照品及样品中异绿原酸3种结构化合物的基线分离,为异绿原酸A、B、C含量的同时测定奠定了基础。该方法优化后缩短了测试时间,为金银花质量的快速检测提供参考依据。

The research aimed to study the optimal chromatographic conditions of the simultaneous determination method of the isochlorogenic acid A, B, and C in flos lonicerae by HPLC. A HPLC method with DAD detector was established and optimized to determine the isochlorogenic acid A, B, and C in Lonicera japonica. The optimal chro-matographic conditions were： Eclipse plus C18 column （4. 6mm×150mm, 5μ m）, with 0. 1 % phosphoric acid （A ） and acetonitrile （B） as mobile phase in gradient mode. The elution conditions were： 0-20min,1 5 % -3 0 % B; 20- 30min, 30% -50% B. The detective wavelength was 325nm, the flow rate was 1. 0mL^min and the column temper-ature was 30℃.The sample volume was 10滋 L. Under the optimal chromatographic conditions, the three structural compound of isochlorogenic acid achieved baseline separation both in reference and sample, which laid the foundation for simultaneous determination of isochlorogenic acid A, B, and C. After optimization, the test time was shortened. This study provides some reference for determination method of rapid detection of Lonicera japonica quality.