Kaale E, Van Goidsenhoven E, Van Schepdael A, Roets E, Hoogmartens J
Laboratory for Pharmaceutical Chemistry and Drug Analysis, K.U. Leuven, Belgium.
Electrophoresis. 2001 Aug;22(13):2746-54. doi: 10.1002/1522-2683(200108)22:13<2746::AID-ELPS2746>3.0.CO;2-2.
This paper describes a system for integration of a one-step-microscale chemical derivatization and analysis by a methodology known as electrophoretically mediated microanalysis (EMMA). Differential electrophoretic mobility between an analyte, reagent, and their product offers EMMA a unique capability to selectively carry out electrophoretic mixing, control product formation, and separation. This system was successfully applied to perform derivatization and separation of the multicomponent aminoglycoside antibiotic gentamicin using 1,2-phthalic dicarboxaldehyde and mercaptoacetic acid as labeling reagents. A multivariate approach based on central composite experimental design was used to optimize the derivative yield. Full automation of the derivatization and analytical procedure, high derivatization efficiency, high sample throughput, and precision are the excellent features of the present method. In addition, this methodology offers short analysis time, as well as selectivity and sensitivity suitable for impurities determination. Separation of gentamicin C1, C1a, C2, C2a, C2b, sisomicin, and several minor components was achieved. For the first time separation and identification of three impurities, namely garamine, 2-deoxystreptamine, and paromamine are described.
本文介绍了一种通过一种称为电泳介导微分析（EMMA）的方法进行一步式微尺度化学衍生化和分析的系统。分析物、试剂及其产物之间的电泳迁移率差异赋予EMMA独特的能力，能够选择性地进行电泳混合、控制产物形成和分离。该系统成功应用于使用1,2-邻苯二甲酸二醛和巯基乙酸作为标记试剂对多组分氨基糖苷类抗生素庆大霉素进行衍生化和分离。基于中心复合实验设计的多变量方法用于优化衍生产率。衍生化和分析过程的完全自动化、高衍生化效率、高样品通量和精密度是本方法的优异特性。此外，该方法分析时间短，具有适合杂质测定的选择性和灵敏度。实现了庆大霉素C1、C1a、C2、C2a、C2b、西索米星和几种次要组分的分离。首次描述了三种杂质即加雷米星、2-脱氧链霉胺和巴龙胺的分离和鉴定。
