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Development of a Poly(dimethylsiloxane) Interface for On-Line Capillary Column Liquid Chromatography: Capillary Electrophoresis Coupled to Sheathless Electrospray Ionization Time-of-Flight Mass Spectrometry
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry, Analytical Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry, Analytical Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry, Analytical Chemistry.
2003 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 75, no 20, 5461-5467 p.Article in journal (Refereed) Published
Abstract [en]

An interface in elastomeric poly(dimethylsiloxane) (PDMS) for on-line orthogonal coupling of packed capillary liquid chromatography (LC) (i.d. = 0.2 mm) with capillary electrophoresis (CE) in combination with sheathless electrospray ionization (ESI) time-of-flight mass spectrometric (TOFMS) detection is presented. The new interface has a two-level design, which in combination with a continuous CE electrolyte flow through the interface provides integrity of the LC effluent and the CE separation until an injection is desired. The transparent and flexible PDMS material was found to have a number of advantages when combined with fused silica column technology, including ease to follow the process and ease to exchange columns. By combining conventional microscale systems of LC, CE, and ESI−MS, respectively, the time scales of the individual dimensions were harmonized for optimal peak capacity per unit time. The performance of the LC−CE−TOFMS system was evaluated using peptides as model substances. A S/N of about 330 was achieved for leucine-enkephaline from a 0.5 μL LC injection of 25 μg/mL peptide standard.

Place, publisher, year, edition, pages
2003. Vol. 75, no 20, 5461-5467 p.
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:uu:diva-90132DOI: 10.1021/ac030117gOAI: oai:DiVA.org:uu-90132DiVA: diva2:162356
Available from: 2003-02-25 Created: 2003-02-25 Last updated: 2011-12-09Bibliographically approved
In thesis
1. Development of Micro Liquid Separation Techniques using Electrospray Ionisation Mass Spectrometry in the Analysis of Polar Compounds and Proteins/Peptides
Open this publication in new window or tab >>Development of Micro Liquid Separation Techniques using Electrospray Ionisation Mass Spectrometry in the Analysis of Polar Compounds and Proteins/Peptides
2003 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Electrospray ionisation (ESI) coupled to mass spectrometry (MS) is one of the most important detection techniques for chemical analysis of small drugs as well as large biomolecules in life science today. In this thesis, aspects on improved compatibility between liquid based separation systems and mass spectrometric detection were investigated regarding buffers, sample preparation and analysis of polar compounds as well as peptides and protein digests for enhanced ESI-MS performance.

Capillary electrophoresis (CE) coupled to ESI-MS detection, was evaluated using both a sheath flow interface and a sheathless design. The separation of peptides and small, polar compounds was optimised for both CE-ESI interfaces. The effect of sheath liquid composition was also studied with the aim to improve sensitivity in the ESI-MS detection.

Polar compounds were retained and separated by capillary ion-pair chromatography coupled to ESI-MS detection. Since commonly used ion-pairing reagents are detrimental to the ESI process they were effectively removed before the ionisation by the use of a trapping column after the separation. Alternatively, the ion-pairing reagents were exchanged to volatile constituents.

A method for peptide mapping by liquid chromatography (LC)-ESI-MS was developed for lactate dehydrogenase. The method was further enhanced to involve the proteolysis on-line to the LC-ESI-MS. No manual sample handling was then needed and the total analysis time decreased from 7 to 1.5 hours. The amount of information was also shown to increase in the on-line system.

Finally, the on-line concept was extended to an innovative interface for direct coupling of a pumped liquid flow to an electroosmotically driven flow. This provided a valve-free sample transfer between capillary LC and CE, aiming towards increased peak capacity per unit time for the analysis of complex peptide samples.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2003. 46 p.
Series
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1104-232X ; 808
Keyword
Analytical chemistry, Analytisk kemi
National Category
Analytical Chemistry
Research subject
Analytical Chemistry
Identifiers
urn:nbn:se:uu:diva-3304 (URN)91-554-5534-4 (ISBN)
Public defence
2003-03-21, B21, Uppsala University, Uppsala, 10:15
Opponent
Available from: 2003-02-25 Created: 2003-02-25 Last updated: 2011-12-09Bibliographically approved

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Bergström, Sara K

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