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Selective Trace Enrichment By Immunoaffinity Capillary Electrochromatography on-Line With Capillary Zone Electrophoresis Laser-Induced Fluorescence

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Abstract:Limited by the lack of a sensitive, universal detector, many capillary-based liquid-phase separation techniques might benefit from techniques that overcome modest concentration sensitivity by preconcentrating large injection volumes. The work presented employs selective solid-phase extraction by immunoaffinity capillary electrochromatography (ACEC) to enhance detection limits. A model analyte, fluorescein isothiocyanate (FITC) biotin, is electrokinetically applied to a capillary column packed with an immobilized anti-biotin-IgG support. After selective extraction by the immunoaffinity capillary, the bound analyte is eluted, migrates by capillary zone electrophoresis (CZE), and is detected by laser-induced fluorescence. The column is regenerated and reused many times. We evaluate the performance of IACEC for selective trace enrichment of analytes prior to CZE. The calibration curve for FITC-biotin bound versus application time is linear from 10 to 300 seconds. Recovery of FITC-biotin spiked into a diluted urinary metabolites solution was 89.4% versus spiked buffer, with a precision of 1.8% relative standard deviation (RSD).
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Citation:Thomas, D. H., D. J. Rakestraw, J. S. Schoeniger, V. Lopezavila, and J. M. Van Emon. Selective Trace Enrichment By Immunoaffinity Capillary Electrochromatography on-Line With Capillary Zone Electrophoresis Laser-Induced Fluorescence. ELECTROPHORESIS 20:57-66, (1999).
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Contact: Liz Hope - (919) 541-2785 or hope.elizabeth@epa.gov
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Division: Human Exposure & Atmospheric Sciences Division
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Branch: Human Expsoure Research Branch
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Product Type: Journal
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Published: 12/31/1999
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Related Entries:
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Bullet Item Field Analytical Methods: Advanced Field Monitoring Methods Development and Evaluation of New and Innovative Technologies That Support the Site Characterization and Monitoring Requirements of the Superfund Program.
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