Sample self-stacking in zone electrophoresis

Gebauer, Petr; Thormann, Wolfgang; Boček, Petr

doi:10.1016/0021-9673(92)87104-g

Cited by 225 publications

(143 citation statements)

References 14 publications

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“…The term sample self-stacking can also be used to characterize a t-ITP preconcentration step induced by the presence of a major component in the sample solution [11].…”

Section: Introductionmentioning

confidence: 99%

Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis

2008

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Electrokinetic supercharging has been integrated in CZE for the development of a highly sensitive methodology for protein tryptic digest analysis. A careful choice of the experimental conditions led to sensitivity enhancement factors between 1000 and 10 000 whilst maintaining a satisfactory resolution. Peptides in the low nanomolar concentration range have been detected despite the use of the poorly sensitive UV absorbance detection mode. The buffer system used in this study is fully suitable for coupling CE to MS.

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“…The term sample self-stacking can also be used to characterize a t-ITP preconcentration step induced by the presence of a major component in the sample solution [11].…”

Section: Introductionmentioning

confidence: 99%

Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis

2008

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“…For biological and environmental samples such as serum, urine and seawater, the presence of high concentrations of chloride is perfect to function as a suitable leading electrolyte for the ITP step, and all that is required to ensure that an appropriate terminator is used and that there is sufficient time to complete the ITP stage and then to destack and separate the components by ZE. Transient ITP (tITP) systems with samples naturally abundant in chloride have been the basis of several studies, the most significant belonging to Gebauer et al for the analysis of biological samples with tITP or originally referred to as sample self stacking [196][197][198][199]. Their first report [196] presented a Electrophoresis 2010, 31, 726-754 simple theory to provide explicit description of various zone parameters such as detection time, variance and resolution, which was confirmed by computer simulation using the GENTRANS software.…”

Section: Aspects Of Ze and Sample Stackingmentioning

confidence: 99%

“…Transient ITP (tITP) systems with samples naturally abundant in chloride have been the basis of several studies, the most significant belonging to Gebauer et al for the analysis of biological samples with tITP or originally referred to as sample self stacking [196][197][198][199]. Their first report [196] presented a Electrophoresis 2010, 31, 726-754 simple theory to provide explicit description of various zone parameters such as detection time, variance and resolution, which was confirmed by computer simulation using the GENTRANS software. This work was later continued to look at the concentrations of components required to induce tITP stacking, with the concentration of the sample macrocomponent or stacking ion required to be above a critical concentration [197], and to study the effect of multiple macro-components upon stacking and conditions under which one of them would function as a destacker [198].…”

Section: Aspects Of Ze and Sample Stackingmentioning

confidence: 99%

Dynamic computer simulations of electrophoresis: A versatile research and teaching tool

et al. 2010

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Software is available, which simulates all basic electrophoretic systems, including moving boundary electrophoresis, zone electrophoresis, ITP, IEF and EKC, and their combinations under almost exactly the same conditions used in the laboratory. These dynamic models are based upon equations derived from the transport concepts such as electromigration, diffusion, electroosmosis and imposed hydrodynamic buffer flow that are applied to user-specified initial distributions of analytes and electrolytes. They are able to predict the evolution of electrolyte systems together with associated properties such as pH and conductivity profiles and are as such the most versatile tool to explore the fundamentals of electrokinetic separations and analyses. In addition to revealing the detailed mechanisms of fundamental phenomena that occur in electrophoretic separations, dynamic simulations are useful for educational purposes. This review includes a list of current high-resolution simulators, information on how a simulation is performed, simulation examples for zone electrophoresis, ITP, IEF and EKC and a comprehensive discussion of the applications and achievements.

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“…Human serum contains a high concentration of salts and this precludes online preconcentration techniques based on conventional sample-stacking or field-enhanced sample injection which require low salt matrices for efficient analyte focusing. Transient ITP or sample self-stacking [29][30][31][32][33][34], formation of a transient moving chemical reaction boundary [35], sweeping [36][37][38], and use of a dynamic pH junction [39], however, are online concentration techniques that can be used to temporarily induce narrow analyte bands in the presence of large amounts of salts. In the present study, electrokinetic sample injection together with sample stacking was studied for high-sensitivity analysis of di-HA and di-CS in model mixtures and human serum digested by chondroitinase ABC using a borate buffer at alkaline pH, reversed electroosmosis (EO) provided by addition of CTAB, and direct UV detection at 232 nm.…”

Section: Introductionmentioning

confidence: 99%

Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking

Yang

Breadmore

Thormann³

2005

J of Separation Science

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Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking CE conditions for monitoring the unsaturated disaccharides of hyaluronic acid (di-HA) and chondroitin sulfate (di-CS) using an alkaline tetraborate buffer, electrokinetic sample injection, and UV absorption detection at 232 nm are reported. Separations were performed in an uncoated fused-silica capillary having reversed polarity and reversed electroosmosis generated with the addition of CTAB to the buffer. The influence of various separation parameters, including the concentration of CTAB, buffer pH, concentration of tetraborate, and applied voltage, on the resolution of the two disaccharides was investigated. Baseline separation was obtained with 25 mM tetraborate at pH 10.0 and having 0.05 mM CTAB. Chloride and phosphate in the sample are beneficial for the stacking of the disaccharides, with di-HA forming a much sharper peak than di-CS. Using samples prepared in 25 mM Tris-HCl (pH 7.5) and electrokinetic injection at the cathode at -10 kV for 40 s, linear relationships between the corrected peak area and the concentration of the disaccharides have been found in the ranges of 1.0-400.0 and 0.1-1.0 lg/mL (0.2-1.0 lg/mL for di-CS), with correlation coefficients being A0.9933 in all cases. The RSDs of detection times and corrected peak areas were between 1.13-1.24 and 1.57-2.13%, respectively. Applied to human serum samples that were prepared by ethanol precipitation and depolymerization of the two polysaccharides with chondroitinase ABC reveals comigration of endogenous compounds with di-HA and a sample-dependent detection time. The di-HA content in the serum sample can be estimated via subtraction of the blank peak that is obtained without enzymatic hydrolysis.

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Sample self-stacking in zone electrophoresis

Cited by 225 publications

References 14 publications

Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis

Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis

Dynamic computer simulations of electrophoresis: A versatile research and teaching tool

Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking

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