胶束电动毛细管色谱法测定硫酸多粘菌素E药物中的多粘菌素E1和E2

摘要

A method of micellar electrokinetic chromatography capillary ( MECC) has been established for separating polymyxins El and E2 in polymyxin E sulfate and determining the contents of El and E2. Several factors including the running voltage, the type of surfactant, concentrations of Bry-35 (polyoxyethyleneglycol dodecyl ether), NaCl solution and acetonitrile, pH of phosphate were investigated. Under the optimum conditions (10 kV running voltage, phos-phate buffer solution (0.01 mol/L, pH4. 1) containing 30 mmol/L Brrj-35, 5% (v/v) acetonitrile, 0. 167 mol/L NaCl), El and E2 were separated with the resolution of 1.94. The contents of El and E2 in polymyxin E sulfate were 67% and 32%, respectively. As an example, the relative standard deviations of the intra-assay and inter-assay of polymyxin El on the plate number and peak area were less than 5%. The method is simple, rapid, accurate, and reproducible.%采用胶束电动毛细管色谱法(MECC)对硫酸多粘菌素E药物中的主要成分多粘菌素E1和E2进行了分离,并测定了多粘菌素E1、E2的含量.分别考察了电泳电压、表面活性剂种类、Brij-35(月桂醇聚氧乙烯醚)浓度、乙腈含量、磷酸盐缓冲液的pH值、氯化钠浓度等实验参数对实验结果的影响,从而确定了最佳的分离条件:电泳电压为10 kV,运行缓冲液为含有30 mmol/L Brij-35、5%(体积分数)乙腈、0.167 mol/L氯化钠的磷酸二氢钠缓冲液(0.01 mol/L,pH 4.1).在优化的实验条件下,E1和E2得到了较好的分离,分离度达到1.94.以多粘菌素E1为例,柱效和峰面积的日间及日内测定的相对标准偏差(RSD)均小于5% .E1和E2在硫酸多粘菌素E药物中的含量分别为67%和32% .该方法简便、快速、准确、重现性好.