On-line coupling of size exclusion chromatography and capillary electrophoresis via solid-phase extraction and a Tee-split interface

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
F W Alexander TempelsGerhardus J de Jong

Abstract

An on-line size exclusion chromatography (SEC)-solid-phase extraction (SPE)-capillary electrophoresis (CE) system using a Tee-split interface has been developed for the analysis of peptides in biological fluids. The SEC column fractionates the sample by molecular size and the low-molecular-weight fraction, which contains the peptides, is directed to a C(18) SPE microcolumn, where the peptides are trapped and concentrated. The SPE column is desorbed with 425 nL acetonitrile and the effluent is sent to the Tee-split interface, which hydrodynamically splits (1:40) the flow and, thus, allows appropriate injection of analytes into the CE system. The performance of the system is investigated by the analysis of enkephalins in cerebrospinal fluid (CSF). It is demonstrated that the SEC step efficiently removes potentially interfering proteins, permitting reproducible SPE and CE. The total system provides efficient separations of the enkephalins with plate numbers up to 100,000. Concentration limits of detection (S/N = 3) for the peptides are about 100 ng/mL for injection of 20 microL spiked CSF samples. Plots of enkephalin peak areas versus concentration showed good linearity over the 0.25-10 microg/mL range (R2 > or = 0.985). Repeatabi...Continue Reading

References

Nov 24, 1999·The Analyst·J R VeraartU A Brinkman
Nov 17, 2004·Journal of Chromatography. a·F W Alexander TempelsGerhardus J de Jong

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Citations

Sep 2, 2008·Analytical and Bioanalytical Chemistry·Anna Poliwoda, Piotr P Wieczorek
May 20, 2008·Analytical Chemistry·Vratislav KostalEdgar A Arriaga
Oct 6, 2009·Analytica Chimica Acta·Maura PerryRobert T Kennedy
Jul 16, 2009·Sensors and Actuators. B, Chemical·Xian HuangQiao Lin
Apr 1, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·David K Lloyd
Nov 24, 2007·Journal of Chromatography. a·Steven L SimpsonShigeru Terabe
Jun 19, 2007·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Patricia PuigGerhardus J de Jong
Nov 30, 2007·Electrophoresis·Václav Kasicka
Dec 7, 2007·Electrophoresis·F W Alexander TempelsGerhardus J de Jong
Sep 18, 2014·Analytical and Bioanalytical Chemistry·Pablo A KlerCarolin Huhn
Oct 16, 2015·Electrophoresis·Rawi RamautarGerhardus J de Jong
Oct 12, 2013·Electrophoresis·Rawi RamautarGerhardus J de Jong

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