Pherogram normalization in capillary electrophoresis and micellar electrokinetic chromatography analyses in cases of sample matrix-induced migration time shifts.
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.
Reijeng
Jetse C
JC
Department of Chemical Engineering and Chemistry, Eindhoven University of Technology, The Netherlands. j.c.reijenga@tue.nl
Martens
Johannes H P A
JH
Giuliani
Andrea
A
Chiari
Marcella
M
D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D004247:DNA / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods*; D015203:Reproducibility of Results; D013996:Time and Motion Studies
D004247:DNA