Isotachophoresis: The concepts of resolution, load capacity and separation efficiency I. Theory

Mikkers, Fep Frans; Everaerts, FM Frans; Peek, Jaf

doi:10.1016/0021-9673(79)80001-1

Cited by 100 publications

(37 citation statements)

References 14 publications

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“…The principles of isotachophoresis have been described by several authors (e.g., HAGLUND, 1970;MIKKERS et al, 1979). In electrophoresis, the charged particles (ions) migrate along with the electric field.…”

Section: Methodsmentioning

confidence: 99%

Isotachophoretic analyses of metabolites of cardiac and skeletal muscles in four species.

et al. 1982

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We studied ATP and concentrations of other metabolites in cardiac and skeletal muscles from different species (frog, hamster, guinea-pig, and dog), using analytical capillary isotachophoresis. The method had several advantages for quantitative analysis of tissue metabolites : short separation time, high sensitivity, high resolution, and good reproducibility. It was also possible to detect a number of compounds simultaneously, including ATP, ADP, AMP, cyclic AMP, creatine phosphate (CP), inosine monophosphate, NAD, NADH, glucose-6-phosphate, pyruvate, and inorganic phosphate. Generally, the concentrations of high-energy phosphates (ATP and CP) in the cardiac muscle were significantly lower than those in the skeletal muscle, in all species tested, except for hamster where the concentration of CP in the skeletal muscle was comparable to that in the cardiac muscle. In mammals, ATP and CP concentrations were comparable in the atrium and ventricle while the concentration in the frog ventricle was significantly higher than that in the atrium. The data obtained by this isotachophoretic analysis were comparable to the data reported by the use of other conventional analytical methods. The significance and reliability of isotachophoresis in the determination of the various metabolites in the cardiac muscle were discussed. Key Words: isotachophoresis, metabolites, cardiac and skeletal muscles.Analytical isotachophoresis is a pertinent tool for the analysis of various chemical compounds and has several advantages, e.g., short separation time, high sensitivity, high resolution, and good reproducibility. With this method, it is possible to detect a large number of metabolites during one single run. This approach has been used to analyze proteins (

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Section: Methodsmentioning

confidence: 99%

Isotachophoretic analyses of metabolites of cardiac and skeletal muscles in four species.

et al. 1982

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“…The use of capillary tubes of larger ids, giving the separation system an enhanced sample loadability [37,42,50], was preferred to work under the conditions providing significantly larger amounts of isolated material in comparison to what can be expected from the current preparative CZE procedures [19,20,24,32,33].…”

Section: Preparative Itp Separations Of Glycosylated Forms Of Rhepomentioning

confidence: 98%

Fractionation of glycoforms of recombinant human erythropoietin by preparative capillary isotachophoresis

et al. 2005

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This feasibility study deals with the use of preparative capillary isotachophoresis (CITP), operating in a discontinuous fractionation mode, to the separations and isolations of glycoforms of recombinant human erythropoietin (rhEPO). The preparative CITP separations were monitored by capillary zone electrophoresis (CZE) with a hydrodynamically closed separation unit. Such a CZE system, suppressing fluctuations of the migration data linked with fluctuations of EOF and hydrodynamic flow, made possible to evaluate and compare the preparative CITP separations performed within a longer time frame. Preparative CITP, carried out in the separation unit with coupled columns of enhanced sample loadability, separating 100 microg of rhEPO in a run lasting ca. 30 min, gave the production rate higher than 55 ng/s for the rhEPO glycoforms. The preparative separations included valve isolations of the glycoforms from the ITP stack into four or six fractions. Such numbers of the fractions corresponded to typical numbers of the major glycoform peaks as resolved in CZE of rhEPO. With respect to close effective mobilities of the glycoforms and a multicomponent nature of rhEPO, the fractions contained mixtures of glycoforms with the dominant glycoforms enriched 10-100-fold, relative to the original rhEPO sample.

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“…In this context, we should note that the preparative ITP runs performed in this study did not represent a limiting production rate of ITP. From a theoretical treatment of the sample load capacity in ITP [22,23] and from some studies dealing with preparative ITP separations [24,25], a further increase of the production rate seems feasible via both enhanced ID of the columns and the concentration of the leading ion.…”

Section: Preparative Itp Separations Of Enantiomersmentioning

confidence: 99%

“…These advantages can be ascribed to the fact that in this combination ITP not only concentrates the enantiomers but also eliminates disturbing matrix constituents from the CE separation compartment (sample clean-up) before a final CZE separation. In a general sense, the analytical power of this combination can be ascribed to an overall clean-up potential of ITP [21] associated with a high sample load capacity of the capillary column in the ITP separation [22,23].…”

Section: Assessment Of Various Capillary Techniques For Preparative Smentioning

confidence: 99%

Separations of enantiomers by preparative capillary isotachophoresis

et al. 1999

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Isotachophoresis: The concepts of resolution, load capacity and separation efficiency I. Theory

Cited by 100 publications

References 14 publications

Isotachophoretic analyses of metabolites of cardiac and skeletal muscles in four species.

Isotachophoretic analyses of metabolites of cardiac and skeletal muscles in four species.

Fractionation of glycoforms of recombinant human erythropoietin by preparative capillary isotachophoresis

Separations of enantiomers by preparative capillary isotachophoresis

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