毛细管电泳法测定硫酸多黏菌素B中的有关物质

Determination of related substances in polymyxin B sulfate by capillary electrophoresis

目的建立一种用毛细管电泳测定硫酸多黏菌素B中有关物质的分析方法。方法以含30mmol/L羟丙基-β-环糊精、5%异丙醇的130mmol/L三乙醇胺溶液(用磷酸调节pH至2.5)为运行缓冲液,熔融石英毛细管总长60cm,有效长度51.5cm,内径50μm,分离电压24kV,柱温25℃,进行了线性、检测限、精密度等方法学考察试验,并将该方法应用于硫酸多黏菌素B原料药及制剂的检测中。结果实现了多黏菌素B中主要组分B1、B2、B3、B1-I与相邻杂质的分离,对比了不同来源硫酸多黏菌素B有关物质的差异。结论本方法灵敏度较高、重复性较好,为硫酸多黏菌素B的质量控制提供了一种可行的分析方法。

Objective A capillary electrophoresis method was established for the determination of related substances in polymyxin B sulfate. Methods To improve the separation selectivity, hydroxypropyl-β-cyclodextrin (HP-β-CD) was necessary to be used as a buffer additive. The optimal separation conditions were as follows: the total length of the capillary was 60cm (the effective length was 51.5cm), 130mmol/L triethanolamine-phosphate buffer containing 30mmol/L HP-β-CD and 5% IPA, the applied voltage was 24kV, and the column temperature was 25℃. Further, the linearity, limits of detection quantification, and the repeatability were examined. The contents of the related substances in polymyxin B of commercial samples were analyzed quantitatively according to the area normalization method. Results The CE method successfully separated the main components (B1, B2, B3, B1-I) and the adjacent impurities in polymyxin B. The contents of the related substances among various sources of polymyxin B were compared. Conclusion The CE method was sensitive and reproducible to control the quality of polymyxin B sulfate.