Yu Hai-Xia, Hao Zeng-Yan, Li Lu, Huang Ya-Yun, Zhang Hong-Fen, Chen An-Jia
Translational Medicine Research Center, Shanxi Medical University, Taiyuan 030001, China.
Pharmacy Department, Shanxi Boai Hospital, Taiyuan 030001, China.
Int J Anal Chem. 2017;2017:3813879. doi: 10.1155/2017/3813879. Epub 2017 Sep 5.
The aim of this study was to develop a method combining an online concentration and high-efficiency capillary electrophoresis separation to analyze and detect three compounds (rutin, hyperoside, and chlorogenic acid) in Flos Farfarae. In order to get good resolution and enrichment, several parameters such as the choice of running buffer, pH and concentration of the running buffer, organic modifier, temperature, and separation voltage were all investigated. The optimized conditions were obtained as follows: the buffer of 40 mM NaHP0-40 mM Borax-30% v/v methanol (pH 9.0); the sample hydrodynamic injection of up to 4 s at 0.5 psi; 20 kV applied voltage. The diode-array detector was used, and the detection wavelength was 364 nm. Based on peak area, higher levels of selective and sensitive improvements in analysis were observed and about 14-, 26-, and 5-fold enrichment of rutin, hyperoside, and chlorogenic acid were achieved, respectively. This method was successfully applied to determine the three compounds in Flos Farfarae. The linear curve of peak response versus concentration was from 20 to 400 g/ml, 16.5 to 330 g/mL, and 25 to 500 g/mL, respectively. The regression coefficients were 0.9998, 0.9999, and 0.9991, respectively.
本研究的目的是开发一种结合在线富集和高效毛细管电泳分离的方法，用于分析和检测款冬花中的三种化合物（芦丁、金丝桃苷和绿原酸）。为了获得良好的分离度和富集效果，对多个参数进行了研究，如运行缓冲液的选择、运行缓冲液的pH值和浓度、有机改性剂、温度和分离电压等。优化后的条件如下：40 mM NaHPO₄-40 mM硼砂-30%（v/v）甲醇缓冲液（pH 9.0）；样品在0.5 psi下进行高达4 s的流体动力学进样；施加20 kV的电压。使用二极管阵列检测器，检测波长为364 nm。基于峰面积，在分析中观察到选择性和灵敏度有更高水平的提高，芦丁、金丝桃苷和绿原酸分别实现了约14倍、26倍和5倍的富集。该方法成功应用于款冬花中三种化合物的测定。峰响应与浓度的线性曲线分别为20至400 μg/ml、16.5至330 μg/mL和25至500 μg/mL。回归系数分别为0.9998、0.9999和0.9991。
