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A hybrid microdevice for electrophoresis and electrochromatography using UV detection

Végvári, Ákos LU and Hjertén, Stellan (2002) In Electrophoresis 23(20). p.3479-3486
Abstract
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... (More)
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--cyclodextrin (allyl--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 (-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 M). 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. (Less)
Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Allyl-beta-cyclodextrin, Electrochromatography, Gel electrophoresis, Glutathione conjugates, Microchip
in
Electrophoresis
volume
23
issue
20
pages
3479 - 3486
publisher
John Wiley & Sons
external identifiers
  • scopus:0036810817
ISSN
0173-0835
DOI
10.1002/1522-2683(200210)23:20<3479::AID-ELPS3479>3.0.CO;2-W
language
English
LU publication?
no
id
0ad97aa4-9e16-424c-9429-3950f9a392d3 (old id 3516233)
date added to LUP
2013-02-25 10:31:03
date last changed
2017-11-19 04:19:14
@article{0ad97aa4-9e16-424c-9429-3950f9a392d3,
  abstract     = {We have designed a new class of microdevices composed of a supporting plastic<br/><br>
(polyvinyl chloride, PVC) plate integrated with a groove for a piece of fused silica capillary<br/><br>
(the separation channel), a slit for on-tube detection, an “islet” for the application<br/><br>
of sample, electrode vessels and platinum electrodes. The design permits electrophoretic,<br/><br>
electrochromatographic and chromatographic separations with on-tube UV<br/><br>
detection. The efficient heat dissipation allows relatively high field strengths. This article<br/><br>
is the first one dealing with microdevices where polymer solutions are replaced by<br/><br>
homogeneous gels. A new type of gels synthesized from acrylamide and 2-hydroxy-3-<br/><br>
allyloxy-propyl--cyclodextrin (allyl--CD) as a cross-linker was employed for electrophoresis<br/><br>
and electrochromatography. 2-Acrylamido-2-methylpropanesulfonic acid was<br/><br>
added to the monomer solution to create a high electroendosmotic flow in electrochromatographic<br/><br>
runs. These gels have excellent electrochromatographic and electrophoretic<br/><br>
properties for low-molecular-weight compounds and DNA, as shown previously,<br/><br>
namely high resolution combined with high stability. The unique cross-linker<br/><br>
can be used for specific interaction with the alkyl and phenyl groups. The tripeptide<br/><br>
glutathione (-L-glutamyl-L-cysteinyl-glycine) and its benzyl conjugates were selected<br/><br>
as model compounds to study the resolving power of the gel because they are difficult<br/><br>
to separate by free zone electrophoresis. The limit of detection (LOD) for S-benzyl-glutathione<br/><br>
was determined (ca. 7 M). Run-by-run reproducibility was high (the separation<br/><br>
factor of glutathione in the gel was 0.3 with 2.5% coefficient of variation, CV). Neutral<br/><br>
compounds (acetone, acetophenone, propiophenone and butyrophenone) were<br/><br>
separated electrochromatographically in the gel. The influence of organic solvent<br/><br>
(acetonitrile) on the electroendosmotic mobility was similar to that in reversed-phase<br/><br>
separations, although the separation mechanism is different. ATP, ADP and AMP were<br/><br>
separated in less than 10 s by free-zone electrophoresis.},
  author       = {Végvári, Ákos and Hjertén, Stellan},
  issn         = {0173-0835},
  keyword      = {Allyl-beta-cyclodextrin,Electrochromatography,Gel electrophoresis,Glutathione conjugates,Microchip},
  language     = {eng},
  number       = {20},
  pages        = {3479--3486},
  publisher    = {John Wiley & Sons},
  series       = {Electrophoresis},
  title        = {A hybrid microdevice for electrophoresis and electrochromatography using UV detection},
  url          = {http://dx.doi.org/10.1002/1522-2683(200210)23:20<3479::AID-ELPS3479>3.0.CO;2-W},
  volume       = {23},
  year         = {2002},
}