Development of a membrane-less dynamic field gradient focusing device for the separation of low-molecular-weight molecules

Jeffrey M Burke; Colin D Smith; Cornelius F Ivory

doi:10.1002/elps.200900589

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Development of a membrane-less dynamic field gradient focusing device for the separation of low-molecular-weight molecules

Journal article

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Development of a membrane-less dynamic field gradient focusing device for the separation of low-molecular-weight molecules

Jeffrey M Burke, Colin D Smith and Cornelius F Ivory

Electrophoresis, Vol.31(5), pp.902-909

03/2010

DOI: https://doi.org/10.1002/elps.200900589

Handle:

https://hdl.handle.net/2376/112472

PMCID: PMC2919354

PMID: 20191553

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Abstract

Electrodes

Water

Fluorescent Dyes - chemistry

Gases

Molecular Weight

Isoelectric Focusing - instrumentation

Isoelectric Focusing - methods

Azo Compounds - isolation & purification

Bromphenol Blue - isolation & purification

Amaranth Dye - isolation & purification

Hydrogen-Ion Concentration

Dynamic field gradient focusing uses an electric field gradient generated by controlling the voltage profile of an electrode array to separate and concentrate charged analytes according to their individual electrophoretic mobilities. This study describes a new instrument in which the electrodes have been placed within the separation channel. The major challenge faced with this device is that when applied voltages to the electrodes are larger than the redox potential of water, electrolysis will occur, producing hydrogen ions (H+) plus oxygen gas on the anodes and hydroxide (OH(-)) plus hydrogen gas on the cathodes. The resulting gas bubbles and pH excursions can cause problems with system performance and reproducibility. An on-column, degassing system that can remove gas bubbles "on-the-fly" is described. In addition, the use of a high capacity, low-conductivity buffer to address the problem of the pH shift that occurs due to the production of H+ on the anodes is illustrated. Finally, the successful separation of three, low-molecular-weight dyes (amaranth, bromophenol blue and methyl red) is described.

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Title: Development of a membrane-less dynamic field gradient focusing device for the separation of low-molecular-weight molecules
Creators: Jeffrey M Burke - Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, WA 99164-2710, USA
Colin D Smith
Cornelius F Ivory
Publication Details: Electrophoresis, Vol.31(5), pp.902-909
Academic Unit: Chemical Engineering and Bioengineering, School of
Publisher: Germany
Grant note: T32 GM008336 / NIGMS NIH HHS TM2GM08336 / NIGMS NIH HHS T32 GM008336-21 / NIGMS NIH HHS T32 GM008336-20 / NIGMS NIH HHS T32 GM008336-19 / NIGMS NIH HHS
Identifiers: 99900547460901842
Language: English
Resource Type: Journal article