Reijeng Jetse C, Martens Johannes H P A, Giuliani Andrea, Chiari Marcella
Department of Chemical Engineering and Chemistry, Eindhoven University of Technology, The Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Apr 25;770(1-2):45-51. doi: 10.1016/s0378-4347(01)00527-8.
When analyzing bio-matrix samples using capillary electrophoresis (CE) or micellar electrokinetic chromatography (MEKC), unwanted shifts in the time axis are often observed, both between samples and standards and between samples, thus hampering identification. These shifts are caused by either or both of two sample matrix-induced effects: variations in stacking conditions (effective field strength or migration length) and variations in electroosmotic flow. Based on elementary CE principles and provided that any two peaks in the pherograms can be linked, these variations can be separately accounted and quantitatively corrected for, so that perfectly overlapping pherograms of standards and samples can be obtained after normalization. The method was validated using samples of a DNA ladder, separated in a sieving polymer. In addition, a number of data files from CE and MEKC analyses (steroids, opioids, beta-blockers, amines, and inorganic anions) previously published by other authors were successfully normalized. A freeware computer programme, CEqualizer, for normalizing ASCII files of detector signals using the method described, is available to the CE community from http: //www.ceyork.f2s.com.
在使用毛细管电泳(CE)或胶束电动色谱法(MEKC)分析生物基质样品时,常常会观察到时间轴上出现不必要的偏移,这种偏移既存在于样品与标准品之间,也存在于不同样品之间,从而妨碍了鉴定。这些偏移是由两种样品基质诱导效应中的一种或两种引起的:堆积条件的变化(有效场强或迁移长度)和电渗流的变化。基于基本的CE原理,并且假设色谱图中的任意两个峰可以关联起来,就可以分别计算并定量校正这些变化,从而在归一化后获得标准品和样品的完美重叠色谱图。该方法通过在筛分聚合物中分离的DNA阶梯样品进行了验证。此外,其他作者先前发表的一些来自CE和MEKC分析(类固醇、阿片类药物、β-受体阻滞剂、胺类和无机阴离子)的数据文件也成功实现了归一化。一个免费的计算机程序CEqualizer,可用于使用上述方法对检测器信号的ASCII文件进行归一化,CE领域的人员可从http://www.ceyork.f2s.com获取该程序。