Church W H, Lee C S, Dranchak K M
Department of Chemistry, Trinity College, Harford, CT 06106-3100, USA.
J Chromatogr B Biomed Sci Appl. 1997 Oct 24;700(1-2):67-75. doi: 10.1016/s0378-4347(97)00314-9.
Addition of cyclodextrins (CDs) to the electrolyte buffer in the capillary zone electrophoresis (CZE) separation of derivatized amino acids was evaluated in terms of fluorescence signal enhancement, resolution, and migration time effects. Maximum fluorescence signal enhancement was observed with separation buffers containing 4 mM beta-cyclodextrin or 10 mM hydroxypropyl beta-cyclodextrin. Resolution values decreased as the CD concentrations increased. Migration times were dependent on CD concentration. Inclusion complex formation constants calculated using changes in migration time showed slight agreement with those calculated by the steady-state fluorescence enhancement technique. Analysis of 20 microl of rat brain microdialysate by CZE using 4 mM beta-cyclodextrin in borate buffer resulted in baseline resolution of glutamate and aspartate in 3.6 min. The results of this work indicate that, when used as separation buffer additives, cyclodextrins are capable of increasing the fluorescence signal and decreasing the migration times of NDA-derivatized acidic amino acids.
在衍生化氨基酸的毛细管区带电泳（CZE）分离中，从荧光信号增强、分离度和迁移时间影响方面评估了向电解质缓冲液中添加环糊精（CDs）的效果。在含有4 mM β-环糊精或10 mM羟丙基-β-环糊精的分离缓冲液中观察到最大荧光信号增强。随着CD浓度增加，分离度值降低。迁移时间取决于CD浓度。使用迁移时间变化计算的包合物形成常数与通过稳态荧光增强技术计算的结果略有一致。在硼酸盐缓冲液中使用4 mM β-环糊精，通过CZE分析20 μl大鼠脑微透析液，在3.6分钟内实现了谷氨酸和天冬氨酸的基线分离。这项工作的结果表明，当用作分离缓冲液添加剂时，环糊精能够增加荧光信号并减少NDA衍生化酸性氨基酸的迁移时间。
