Underberg Willy J M, Waterval Joop C M
Department of Biomedical Analysis, Faculty of Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
Electrophoresis. 2002 Nov;23(22-23):3922-33. doi: 10.1002/elps.200290010.
This survey as a sequel of two earlier reports gives an overview of recent developments, starting from 1999, in the use of derivatization protocols in capillary electrophoretic (CE) analysis. Derivatization is mainly used for enhancement of the detection sensitivity in CE, for which a combination of fluorescence labeling and laser-induced fluorescence detection is favorable. Moreover, especially in the field of saccharide assay, derivatization to introduce charge into the molecule, is common. Derivatization procedures are classified in tables, focused on precapillary, on-line, on-capillary and postcapillary arrangements and divided in sections concerning the functional group that is derivatized. The most frequently reported groups are amines and the reducing end of (oligo)saccharides, but thiols, carbonyl and carboxyl groups, steroids and inorganic ions have also been reported about. Other reasons for derivatization are to enhance chiral separation, introduction of a suitable charge into the molecule or to improve mass spectrometric detection. The use of derivatization techniques for special cases, such as the analysis of neurotransmitters, insulin antibodies and mitochondria has also been incorporated as well as a study on the adsorption of proteins onto capillary walls during CE in which derivatization plays a role.
本次调查作为前两份报告的续篇,概述了自1999年以来毛细管电泳(CE)分析中衍生化方法的最新进展。衍生化主要用于提高CE检测灵敏度,荧光标记与激光诱导荧光检测相结合对此较为有利。此外,特别是在糖类分析领域,通过衍生化给分子引入电荷很常见。衍生化方法在表格中分类,重点是柱前、在线、柱上和柱后配置,并按衍生化的官能团分为不同部分。最常报道的基团是胺和(寡)糖的还原端,但也有关于硫醇、羰基和羧基、类固醇及无机离子的报道。衍生化的其他原因包括增强手性分离、给分子引入合适电荷或改善质谱检测。特殊情况下衍生化技术的应用,如神经递质分析、胰岛素抗体分析和线粒体分析也被纳入,还有一项关于CE过程中蛋白质在毛细管壁上吸附的研究,其中衍生化发挥了作用。