分散微固相萃取-高效液相色谱法测定荷丹片中生物碱类成分的含量

Dispersive Micro Solid-Phase Extraction Coupled with HPLC-UV for Detection of Nuciferine,N-nornuciferine,O-nornuciferine in He-Dan-Pian

目的建立分散微固相萃取-高效液相色谱法测定荷丹片中荷叶碱、O-去甲基荷叶碱、N-去甲基荷叶碱的含量的新方法。方法以强离子交换树脂(PCX)为吸附剂,通过对DMSPE技术中PCX用量、吸附时间、洗脱溶剂氨-乙腈浓度和洗脱体积的优化,实现样品中荷叶碱的最优提取纯化。结果建立了测定荷丹片中荷叶碱、O-去甲基荷叶碱、N-去甲基荷叶碱含量的分散微固相萃取-高效液相色谱法,分散微固相萃取的最佳工艺条件为:PCX用量25 mg,涡旋30 s,以3.0 mL 5%的氨-乙腈溶液进行洗脱。结论建立的含量测定方法可用于荷丹片中荷叶碱、O-去甲基荷叶碱、N-去甲基荷叶碱含量的分析测定。该方法简单精确,稳定性高;样品处理快捷简便,溶剂损耗明显减少;设备要求低,装置简便;分析速度快,分析周期短。

Objective To establish a HPLC method for the determination of nuciferine, N-nornuciferine, O-nornuciferine in a traditional Chinese medicine He-Dan-Pian coupled with dispersive micro solid-phase extraction(DMSPE), using polymer cation exchange(PCX) material as an adsorbent. Methods The main factors affecting the extraction efficiency(the amount of adsorbent, the adsorption time, the type and volume of eluent) were investigated in detail. Results The DMSPE-HPLC method for determination of nuciferine, N-nornuciferine, O-nornuciferine in He-Dan-Pian was established. The optimum conditions of DMSPE were as follows: the amount of PCX was 25 mg, adsorption time was 30 s, the elute solution was 5% ammonia-acetonitrile and the volume of elute solution was 3.0 mL. ConclusionUsing this DMSPE-HPLC method, the content of nuciferine, N-nornuciferine, O-nornuciferine in He-Dan-Pian can be measured. Compared with previously published liquid-liquid extraction method, this new method can significantly improve the selectivity and sensitivity of the analytes. As well as, the validated method afforded desired linearity, precision, accuracy, sensitivity and stability. This new cleanup method is expected to be applied for the analysis of bioactive compounds in traditional Chinese medicine in the future.