Chromatographic preconcentration coupled on-line to capillary electrophoresis via a Tee-split interface

Tempels, F. W. Alexander; Teeuwsen, J.; Kyriakou, Ioannis K.; Theodoridis, Georgios; Underberg, W.J.M.; Somsen, Govert W.; Jong, Gerhardus J. de

doi:10.1016/j.chroma.2004.07.042

Cited by 22 publications

(15 citation statements)

References 31 publications

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“…A 1000-fold sensitivity enhancement in comparison with the direct sample application was achieved in analysis of enkephalin peptides with detection limits of 5-10 ng/mL. Alternatively, the chromatographic concentration is connected online to CE via a Tee-split interface [35].…”

Section: Preconcentration and Preseparationmentioning

confidence: 99%

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

2006

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The article gives a comprehensive review on the recent developments in the applications of high-performance capillary electromigration methods, zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography, to analysis, preparation, and physicochemical characterization of peptides. The article presents new approaches to the theoretical description and experimental verification of electromigration behavior of peptides, covers the methodological aspects of capillary electroseparations of peptides, such as rational selection of separation conditions, sample preparation, suppression of peptide adsorption, new developments in individual separation modes, and new designs of detection systems. Several types of applications of capillary electromigration methods to peptide analysis are presented: conventional qualitative and quantitative analysis, purity control, determination in biomatrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid and sequence analysis, and peptide mapping of proteins. Some examples of micropreparative peptide separations are given and capabilities of capillary electromigration techniques to provide important physicochemical characteristics of peptides are demonstrated.

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Section: Preconcentration and Preseparationmentioning

confidence: 99%

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

2006

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“…Without changing analytical performance (e.g., due to the use of a different column), the nanoLC can be accelerated by using column-switching techniques for faster injection. The same approach is usable for automation and acceleration of the large volume CE injection using a Tee-split interface as recently demonstrated by Tempels et al [42].…”

Section: Time Requestsmentioning

confidence: 74%

Separation techniques hyphenated to electrospray‐tandem mass spectrometry in proteomics: Capillary electrophoresis versus nanoliquid chromatography

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2005

Electrophoresis

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Liquid chromatography (LC) nanoelectrospray-tandem mass spectrometry (MS/MS) is a key technology for the study of proteomics, with the main benefit to the characterization of sensitive peptides from complex mixtures. Capillary electrophoresis coupled to mass spectrometry (MS) has been taken into consideration sporadically due to the highly efficient separation and ability to handle low sample amount, yet classified as being less sensitive with respect to analyte concentration. The limitation in capillary zone electrophoresis (CZE) injection volumes can be overcome by on-line solid-phase extraction (SPE). Such an on-line SPE-CZE system was explored in combination with an ion trap (IT) mass spectrometer. Thus, it was possible to inject more than 100 microL sample solution on to the CZE capillary. Concentration limits of detection as low as 100 amol/microL were demonstrated for a peptide standard. This SPE-CZE-microelectrospray ionization (ESI)-MS/MS setup was compared directly to nanoLC/nanoESI using the same sample of a tryptic digest of bovine serum albumin (BSA) as a reference standard. Measurements were made on one IT mass spectrometer with identical acquisition parameters. Both chromatography systems enabled the separation and detection of low levels of peptides from a mixture of moderate complexity, with most peptides identified using both techniques; however, specific differences were obvious. The nanoLC-MS is about five times more sensitive than the CZE-MS, yet the difference was less pronounced than expected. The CZE-MS technique showed reduced loss of peptides, especially for larger peptides (missed cleavages) and is about four times faster than the nanoLC-MS approach.

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“…On-line chromatographic preconcentration with CE is promising due to the large sample enrichment capacity [30,31]; however, it is technically challenging to couple effectively since conditions used for analyte elution often result in significant band-broadening that can deteriorate overall separation performance. In contrast, on-line sample preconcentration by dynamic pH junction in CE represents a simple method to electrokinetically focus large sample volumes of various weakly ionic metabolites directly in-capillary while retaining high resolution [2].…”

Section: Integration Of On-line Preconcentration With Chemical Derivamentioning

confidence: 99%

On-line sample preconcentration with chemical derivatization of bacterial biomarkers by capillary electrophoresis: A dual strategy for integrating sample pretreatment with chemical analysis

2005

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Simple, selective yet sensitive methods to quantify low-abundance bacterial biomarkers derived from complex samples are required in clinical, biological, and environmental applications. In this report, a new strategy to integrate sample pretreatment with chemical analysis is investigated using on-line preconcentration with chemical derivatization by CE and UV detection. Single-step enantioselective analysis of muramic acid (MA) and diaminopimelic acid (DAP) was achieved by CE via sample enrichment by dynamic pH junction with ortho-phthalaldehyde/N-acetyl-L-cysteine labeling directly in-capillary. The optimized method resulted in up to a 100-fold enhancement in concentration sensitivity compared to conventional off-line derivatization procedures. The method was also applied toward the detection of micromolar levels of MA and DAP excreted in the extracellular medium of Escherichia coli bacterial cell cultures. On-line preconcentration with chemical derivatization by CE represents a unique approach for conducting rapid, sensitive, and high-throughput analyses of other classes of amino acid and amino sugar metabolites with reduced sample handling, where the capillary functions simultaneously as a concentrator, microreactor, and chiral selector.

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Chromatographic preconcentration coupled on-line to capillary electrophoresis via a Tee-split interface

Cited by 22 publications

References 31 publications

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

Separation techniques hyphenated to electrospray‐tandem mass spectrometry in proteomics: Capillary electrophoresis versus nanoliquid chromatography

On-line sample preconcentration with chemical derivatization of bacterial biomarkers by capillary electrophoresis: A dual strategy for integrating sample pretreatment with chemical analysis

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