Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, Shanghai, PR China.
J Sep Sci. 2010 Apr;33(8):1099-108. doi: 10.1002/jssc.200900657.
A rapid CEC method with poly(butyl methacrylate-co-ethylene dimethacrylate-co-[2-(methacryloyloxy)ethyl] trimethylammonium chloride) monolithic column has been developed for separation and determination of four coumarins (isopimpinelline, bergapten, imperatorin, and osthole) in Fructus cnidii extracts. The effect of polymerization condition including the monomers ratio and the porogens ratio were studied. The mobile-phase composition, such as the composition of organic solvent, the concentration and pH of buffer, was also optimized. Under the same condition (50% ACN and 50% of a 10 mM sodium dihydrogen phosphate electrolyte at pH 4.95), in contrast to 25 min of analysis time in HPLC and 10 min of analysis time in pCEC, a fast separation of these analytes was achieved in less than 5 min in CEC. Method validation was developed in accordance with the analytical procedures. Intra- and interday precisions (RSD) for relative retention time and peak area were less than 1.69 and 4.63%, and LODs were lower than 0.5 microg/mL. Calibration curves of four compounds also showed good linearity (r(2)>0.995). The mean recoveries ranged between 93.91 and 98.65%. With this CEC system, the quality of F. cnidii extracts from various resources was evaluated by determining the contents of the four coumarins.
已开发出一种快速 CEC 方法，使用聚（甲基丙烯酸丁酯-甲基丙烯酸二乙稀基酯-（2-（甲基丙烯酰氧基）乙基）三甲基氯化铵）整体柱分离和测定蛇床子提取物中的四种香豆素（异茴芹素、佛手柑内酯、花椒毒素和蛇床子素）。研究了聚合条件（单体比例和致孔剂比例）的影响。还优化了流动相组成，如有机溶剂组成、缓冲液浓度和 pH 值。在相同条件下（50% ACN 和 50% 10 mM 磷酸二氢钠电解质，pH 值 4.95），与 HPLC 分析时间 25 分钟和 pCEC 分析时间 10 分钟相比，这些分析物在 CEC 中不到 5 分钟即可快速分离。根据分析程序开发了方法验证。相对保留时间和峰面积的日内和日间精密度（RSD）小于 1.69%和 4.63%，LODs 低于 0.5 μg/mL。四种化合物的校准曲线也表现出良好的线性（r²>0.995）。平均回收率在 93.91%至 98.65%之间。使用该 CEC 系统，通过测定四种香豆素的含量来评估来自不同资源的蛇床子提取物的质量。
