On-line coupling of size exclusion chromatography and capillary electrophoresis via solid-phase extraction and a Tee-split interface - Analytical Methods & Tools for Lyophilization

On-line coupling of size exclusion chromatography and capillary electrophoresis via solid-phase extraction and a Tee-split interface
Engineering Village 2
? 2006 Elsevier Inc
Accession number: 062910008033

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

Authors: Tempels, F.W. Alexander; Wiese, Gerard; Underberg, Willy J.M.; Somsen, Govert W.; de Jong, Gerhardus J.

Author affiliation: Department of Biomedical Analysis, Faculty of Pharmaceutical Sciences, Utrecht University, NL-3584 CA Utrecht, Netherlands

Serial title: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

Abbreviated serial title: J. Chromatogr. B Anal. Technol. Biomed. Life Sci.

Volume: v 839

Issue: n 1-2

Issue date: Jul 24 2006

Publication year: 2006

Pages: p 30-35

Language: English

ISSN: 1570-0232

CODEN: JCBAAI

Document type: Journal article (JA)

Publisher: Elsevier, Amsterdam, 1000 AE, Netherlands

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 C18 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 ?L spiked CSF samples. Plots of enkephalin peak areas versus concentration showed good linearity over the 0.25-10 ?g/mL range (R2 greater than or equal 0.985). Repeatability of migration time and peak area was within 2% and 10% R.S.D., respectively. ? 2006 Elsevier B.V. All rights reserved.

Number of references: 21

Ei main heading: Amino acids

Ei controlled terms: Molecular weight - Acetonitrile - Hydrodynamics - Size exclusion chromatography - Electrophoresis - Separation

Uncontrolled terms: Solid-phase extraction - Capillary electrophoresis - Tee split interface - Enkephalin - Cerebrospinal fluid

Ei classification codes: 804.1 Organic Compounds - 931.3 Atomic & Molecular Physics - 631.1.1 Liquid Dynamics - 801 Chemistry - 701.1 Electricity: Basic Concepts & Phenomena - 802.3 Chemical Operations

Treatment: Theoretical (THR); Experimental (EXP)

DOI: 10.1016/j.jchromb.2006.02.003

Database: Compendex

Compilation and indexing terms, ? 2006 Elsevier Inc. All rights reserved
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