High performance capillary electrophoresis separation modes

Suzuki, S., Kakehi, K., and Honda, S., Two-dimensional mapping of N-gly-cosidically linked asialo-oligosaccharides from glycoproteins as reductively pyridylaminated derivatives using dual separation modes of high-performance capillary electrophoresis, Anal. Biochem., 205, 227, 1992. 

Fan et al. [106] developed a high performance capillary electrophoresis method for the analysis of primaquine and its trifluoroacetyl derivative. The method is based on the mode of capillary-zone electrophoresis in the Bio-Rad HPE-100 capillary electrophoresis system effects of some factors in the electrophoretic conditions on the separation of primaquine and trifluoroacetyl primaquine were studied. Methyl ephedrine was used as the internal standard and the detection was carried out at 210 nm. A linear relationship was obtained between the ratio of peak area of sample and internal standard and corresponding concentration of sample. The relative standard deviations of migration time and the ratio of peak area of within-day and between-day for replicate injections were <0.6% and 5.0%, respectively. 

At times, capillary electrophoresis is called high-performance capillary electrophoresis to reflect the high number of theoretical plates offered by this technique as a result, superior resolution is possible. The original contribu-tor(s) to high-performance capillary electrophoresis and the basis of separation in various modes are shown in Table 2.4 9... 

The application of high-performance capillary zone electrophoresis (HP-CZE) in its various selectivity modes has become a very valuable adjunct to HPLC for the analysis of peptides. For synthetic peptides, in particular, both HPLC and HP-CZE now form essential components of the analytical characterization of these molecules. Increasingly, zonal, micellar, or (biospecific) affinity-based HP-CZE procedures with open tubular capillary systems are adapted to allow resolution with extremely high separation efficiencies (e.g., >105 plates per meter) of synthetic or naturally occurring peptides as part of the determination of their structural, biophysical, or functional properties. Illustrative of these capabilities are the results shown in Figure 19 for the separation of several peptides with different charge and Stokes radius characteristics by HP-CZE. 

Identification and quantification of natural dyes need high performance analytical techniques, appropriate for the analysis of materials of complicated matrices containing a small amount of coloured substances. This requirement perfectly fits coupling of modern separation modules (usually high performance liquid chromatography in reversed phase mode, RPLC, but also capillary electrophoresis, CE) with selective detection units (mainly mass spectrometer). 

There are several good reasons why CE has not captured the lion s share of small-molecule separations. High-performance liquid chromatography (HPLC) has a 23-year head start over capillary electrophoresis, and most of the problems have been worked out. HPLC is rugged, sensitive, scales up to preparative and commercial modes, and scales down to the capillary format. Poorly developed CE methods by ill-trained chromatographers are another contributing factor to the slow acceptance of CE in the world of small molecules. 

Mass spectrometry (MS) possesses the unique abihty to identify separated compounds and is now used routinely with gas and liquid chromatography. This selective and highly sensitive detection mode is also compatible with capillary electrophoresis and provides a powerful combination for performing rapid, efficient and sensitive analysis. 

During the last decades the different modes of capillary electrophoresis (CE) have developed into high-performance (HPCE) separation methods, offering a high separation efficiency (lO -lO theoretical plates), high sensitivity (femtomole to zeptomole amounts in nano- to picoliter sample volumes), and short analysis times (typically 5-20 min, in special cases only a few seconds). They are considered as a recognized complement and/or coimterpart of liquid chromatography (LC) and gel electrophoretic methods. 

Owing to its outstanding high-performance separation potential also capillary electrophoresis has been employed in different modes and with different detection methods for separation and determination of MC. Both capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC), which employs a surfactant above the critical micelle concentration to separate neutral and uncharged molecules based on electromigration principle, have been employed for determination of... 

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