Background electrolyte zones

Indirect UV absorbance detection in capillary zone electrophoresis has been used to analyze sodium alcohol sulfates. Excellent reproducibility was obtained when veronal buffer was used as UV-absorbing background electrolyte [302],... 

Both types of units have generally been operated in trace mode that is, background or elutant electrolyte is fed to the unit along with the mixture to be separated. A desirable and possible means of operation for preparative applications is in bulk mode, in which one separated component follows the other without background electrolyte being present, except that other ions may be required to bracket the separated zones. Overlap regions between components should be recycled, and pure components collected as products. 

Capillary electrophoresis (CE) or capillary zone electrophoresis (CZE) is the technique most often employed in pesticide residue analysis. In its most basic form, free zone electrophoresis, a fused-silica capillary is filled with electrolyte (running buffer or background electrolyte). A potential is applied across the capillary and the cations... 

Fig. 3.161. (A) Zone electrophoresis patterns of FITC-labelled transferrin samples by fluorescence detection. The unbound dye (providing a main peak and several minor ones) was not removed from the samples. Experimental conditions background electrolyte, 100 mM borate buffer, pH 8.3 voltage, 20 kV capillary 59 cm (effective length 41 cm) X 75 pm i.d. injection of samples 100 mbar x s 20°C detection with fluorescence detector (240 - 400 nm, broadband excitation filter and a 495 nm cut-off emmision filter). The reaction was left to continue for 20 h, and the reaction mixtures contained 13 pm (1 mg/ml) Tf and (a) 0.01 mM FITC, (b) 0.1 mM FITC, and 1 mM FITC. (B) Zone electrophoresis patterns of an FITC-labelled transferrin sample by simultaneous fluorescence (upper trace, left axis) and UV detection (lower trace, right axis). The unbound dye shows several peaks with both detections. Experimental conditions background electrolyte, 100 mM borate buffer, pH 8.3 voltage, 20 kV capillary 59 cm (effective length fluorescence 41 cm, UV 50.5 cm) X 75 pm i.d. injection of samples 100 mbar X s 20°C detection with fluorescence detector (240 - 400 nm, broadband excitation filter and a 495 nm cut off emmision filter). The reaction was left to continue for 20 h, and the reaction mixtures contained 6.5 pm (0.5 mg/ml) Tf and 0.1 mM FITC. Reprinted with permission from T. Konecsni et al. [199].

In CZE, the capillary, inlet reservoir, and outlet reservoir are filled with the same electrolyte solution. This solution is variously termed background electrolyte, analysis buffer, or run buffer. In CZE, the sample is injected at the inlet end of the capillary, and components migrate toward the detection point according to their mass-to-charge ratio by the electrophoretic mobility and separations principles outlined in the preceding text. It is the simplest form of CE and the most widely used, particularly for protein separations. CZE is described in Capillary Zone Electrophoresis. ... 

FIGURE 5 Schematic representation of the mechanism for enantiomeric separation in chiral CE of basic compounds with cyciodextrin type selectors. The model electropherograms represent I blank run with buffer electrolyte at acidic pH 2 sample run with buffer electrolyte at acidic pH, no enantiomeric separation is observed 3 blank run with background electrolyte including a selector, e.g., cyciodextrin. Note a small delay in the EOF zone and 4 sample run with background electrolyte containing a selector, e.g., cyciodextrin, resulting in enantiomeric separation of the peaks. 

Beckers, J. L., and Bocek, P. (2003). The preparation of background electrolytes in capillary zone electrophoresis golden rules and pitfalls. Electrophoresis 24, 518 — 535. 

Jaros, M., Hruska, V., Stedry, M., Zuskova, I., and Gas, B. (2004). Eigenmobilities in background electrolytes for capillary zone electrophoresis. IV. Computer program PeakMaster. Electrophoresis 25, 3080-3085. 

Figure 26-25 Stacking of anions and cations at opposite ends of the low-conductivity sample plug (zone) occurs because the electric field in the sample plug is much higher than the electric field in the background electrolyte. Time increases from panels a to d. Electroneutrality is maintained by migration of background electrolyte ions, which are not shown.

E Electrochromatography. It is defined by Heftmann(Ref 78,p 14) as "a method of analysis in which direct current electrical potential promotes the separation of substances by differential migration from a narrow zone in a stabilized background electrolytic solution . 

In capillary zone electrophoresis microchips, where the background electrolyte consists only of aqueous buffer, analytes are separated based on a size-to-charge ratio, and neutral analytes are not resolved from each other. 

Berzas Nevado et al. [138] developed a new capillary zone electrophoresis method for the separation of omeprazole enantiomers. Methyl-/ -cyclodextrin was chosen as the chiral selector, and several parameters, such as cyclodextrin structure and concentration, buffer concentration, pH, and capillary temperature were investigated to optimize separation and run times. Analysis time, shorter than 8 min was found using a background electrolyte solution consisting of 40 mM phosphate buffer adjusted to pH 2.2, 30 mM /1-cyclodextrin and 5 mM sodium disulfide, hydrodynamic injection, and 15 kV separation voltage. Detection limits were evaluated on the basis of baseline noise and were established 0.31 mg/1 for the omeprazole enantiomers. The method was applied to pharmaceutical preparations with recoveries between 84% and 104% of the labeled contents. 

The most common classification scheme in electrophoresis focuses on the nature of electrolyte system. Using this scheme, electrophoretic modes are classified as continuous or discontinuous systems. Within these groupings the methods may be further divided on the basis of constancy of the electrolyte if the composition of the background electrolyte is constant as in capillary zone electrophoresis, the result is a kinetic process. If the composition of the electrolyte is not constant, as in isoelectric focusing, the result is a steady-state process. 

Most CE work so far has been done using the capillary zone electrophoresis (CZE) mode, where analytes are separated on the basis of differences in electrophoretic mobility, which is related to charge density. The separation is carried out in a capillary filled with a continuous background electrolyte (buffer). Micellar electrokinetic capillary chromatography (MEKC or MECC) is one other CE method based on differences in the interaction of the analytes with micelles present in the separation buffer, which can easily separate both charged and neutral solutes with either hydrophobic or hydrophilic properties. An alternative to MEKC is capillary... 

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