A hybrid microdevice for electrophoresis and electrochromatography using UV detection
Författare
Summary, in English
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.
(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.
Publiceringsår
2002
Språk
Engelska
Sidor
3479-3486
Publikation/Tidskrift/Serie
Electrophoresis
Volym
23
Issue
20
Länkar
Dokumenttyp
Artikel i tidskrift
Förlag
John Wiley & Sons Inc.
Ämne
- Medical Engineering
Nyckelord
- Allyl-beta-cyclodextrin
- Electrochromatography
- Gel electrophoresis
- Glutathione conjugates
- Microchip
Status
Published
ISBN/ISSN/Övrigt
- ISSN: 0173-0835