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.
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.
参考文献
| [1] | Wang X,Xiang W L,Feng W.Modern Agricultural Sciences (王旭,向文良,冯玮.现代农业科学),2009,16(1):24 |
| [2] | Zhao H Q,Zhong J M,Feng J,et al.Guizhou Animal Science and Veterinary Medicine (赵海全,钟健敏,冯军,等.贵州畜牧兽医),2007,31(3):7 |
| [3] | Suzuki T,Inouye H,Fujikawa K,et al.J Biochem,1963,54:25 |
| [4] | British Pharmacopoeia Commission Office.British Pharmacopoeia 2009.London:Stationary Office,2009:1 594 |
| [5] | Thomas A H,Thomas J M,Hollway I.Analyst,1980,105:1 068 |
| [6] | Kitagawa T,Ohtani W,Maeno Y,et al.J Assoc Off Chem,1985,68:661 |
| [7] | Grande B C,Falcon S M G,Perez-Lamela C,et al.Chromatographia (Suppl),2001,53:S460 |
| [8] | European Pharmacopoeia Commission.European Pharmacopoeia 5th ed.Strasbourg:Council of Europe,2005:1 361 |
| [9] | Zhang H W,Hu C Q,Xu M Z,et al.Chinese Journal of Chromatography (张惠文,胡昌勤,许明哲,等.色谱),2007,25(9):699 |
| [10] | Gan N,Li T H,Wang L Y,et al.Chinese Journal of Chromatography (干宁,李天华,王鲁雁,等.色谱),2007,25(11):934 |
| [11] | Chen Y.Technology and application of capillary electrophoresis.Beijing:Chemical Industry Press (陈义.毛细管电泳技术及应用.北京:化学工业出版社),2000:48 |
- 下载量()
- 访问量()
- 您的评分：
-
10%
-
20%
-
30%
-
40%
-
50%