uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
A Hybrid Microdevice for Electrophoresis and Electrochromatography Using UV-Detection
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry.
2002 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 23, no 20, 3479-3486 p.Article in journal (Refereed) Published
Abstract [en]

We have designed a new class of microdevices composed of a supporting plastic (polyvinyl chloride, PVC) plate integrated with a groove for a piece of fused silica capillary (the separation channel), a slit for on-tube detection, an "islet" for the application of sample, electrode vessels and platinum electrodes. The design permits electrophoretic, electrochromatographic and chromatographic separations with on-tube UV detection. The efficient heat dissipation allows relatively high field strengths. This article is the first one dealing with microdevices where polymer solutions are replaced by homogeneous gels. A new type of gels synthesized from acrylamide and 2-hydroxy-3-allyloxy-propyl-beta-cyclodextrin (allyl-beta-CD) as a cross-linker was employed for electrophoresis and electrochromatography. 2-Acrylamido-2-methylpropanesulfonic acid was added to the monomer solution to create a high electroendosmotic flow in electrochromatographic runs. These gels have excellent electrochromatographic and electrophoretic properties for low-molecular-weight compounds and DNA, as shown previously, namely high resolution combined with high stability. The unique cross-linker can be used for specific interaction with the alkyl and phenyl groups. The tripeptide glutathione (gamma-L-glutamyl-L-cysteinyl-glycine) and its benzyl conjugates were selected as model compounds to study the resolving power of the gel because they are difficult to separate by free zone electrophoresis. The limit of detection (LOD) for S-benzyl-glutathione was determined (ca. 7 microM). Run-by-run reproducibility was high (the separation factor of glutathione in the gel was 0.3 with 2.5% coefficient of variation, CV). Neutral compounds (acetone, acetophenone, propiophenone and butyrophenone) were separated electrochromatographically in the gel. The influence of organic solvent (acetonitrile) on the electroendosmotic mobility was similar to that in reversed-phase separations, although the separation mechanism is different. ATP, ADP and AMP were separated in less than 10 s by free-zone electrophoresis.

Place, publisher, year, edition, pages
2002. Vol. 23, no 20, 3479-3486 p.
National Category
Natural Sciences
URN: urn:nbn:se:uu:diva-89960DOI: 10.1002/1522-2683(200210)23:20<3479::AID-ELPS3479>3.0.CO;2-WPubMedID: 12412115OAI: oai:DiVA.org:uu-89960DiVA: diva2:161917
Available from: 2002-09-13 Created: 2002-09-13 Last updated: 2013-06-12Bibliographically approved
In thesis
1. True Monoliths as Separation Media: Homogeneous Gels for Electrophoresis and Electrochromatography in the Capillary and Microchip Modes
Open this publication in new window or tab >>True Monoliths as Separation Media: Homogeneous Gels for Electrophoresis and Electrochromatography in the Capillary and Microchip Modes
2002 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The thesis focuses on the development of new homogeneous gels for the separation of drug enantiomers, peptides, DNA and virus by electrophoresis and electrochromatography in capillaries and microchips. This type of separation media offers high resolution and small zone broadening. Compared to particulate beds the resolution in this type of separation media is high because the eddy diffusion is zero and the resistance to mass transfer is small, since the diffusional distance between two polymer chains in the gel is considerably shorter than that between two beads in a packed bed.

The gels have been characterized in terms of plate heights, plate numbers, resolution, etc. Gels of agarose, polyvinyl alcohol, albumin and polyacrylamide have been employed for electrochromatography or electrophoresis. N,N’-methylene-bisacrylamide, the most widely used crosslinker in polyacrylamide gels, was exchanged for allyl-β-cyclodextrin to get a multi-purpose gel, i.e., a separation medium the separation properties of which is determined not only by the polyacrylamide chains, but also by β-cyclodextrin with its complexation power.

A cost-effective, hybrid microdevice has been designed for fast electrophoretic and electrochromatographic analyses as well as for microchromatography. It consists of a fused silica capillary mounted on a supporting plate which integrates most of the compartments necessary for automation and sensitive detection at short UV wavelengths.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2002. 46 p.
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1104-232X ; 747
Biochemistry, capillary electrophoresis, capillary electrochromatography, gel electrophoresis, agarose, polyacrylamide, b-cyclodextrin, DNA, peptide, glutathione, drug enantiomers, microchip, Biokemi
National Category
Biochemistry and Molecular Biology
Research subject
urn:nbn:se:uu:diva-2584 (URN)91-554-5395-3 (ISBN)
Public defence
2002-10-04, Lecture Hall B21, BMC, Uppsala, 10:00
Available from: 2002-09-13 Created: 2002-09-13Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMed

Search in DiVA

By author/editor
Hjertén, Stellan
By organisation
Department of Biochemistry
In the same journal
Natural Sciences

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 215 hits
ReferencesLink to record
Permanent link

Direct link