Peroxyoxalate Chemiluminescence Detection for Capillary Electrophoresis Using Membrane Collection

Loranelle L. Shultz, Scott Shippy, Timothy A. Nieman, Jonathan V. Sweedler

Research output: Contribution to journalArticle

Abstract

Capillary electrophoresis (CE) is often limited by the need to measure nanoliter-volume, low-concentration analytes with selectivity and specificity. A post-column chemiluminescence (CL) method is described in which the capillary effluent is collected on a membrane which has been coated with a peroxyoxalate CL reagent, bis(2,4,6-trichlorophenyl)oxalate. After the CE run, the CL reaction is initiated on the membrane by applying hydrogen peroxide, a scientific charge-coupled device (CCD) camera images the resultant CL emission, and an electropherogram is reconstructed. This postcolumn detection scheme decouples the separation parameters from detection and allows longer lived CL reagents to be used with fast separations. As the reaction is catalyzed by imidazole, CL intensities and lifetimes resulting from imidazole- and non-imidazole-running buffers are compared. The postcolumn CL method is compared to on-line UV absorbance detection for a mixture of dansylated amino acids, with limits of detection (LODs) of 1.5 μM (13 fmol) and separation efficiencies of 170,000 plates. The ability to preserve the sample on the membrane allows several selective detection schemes to be employed; the combination of postcolumn chemiluminescence and radionuclide detection from a single electrophoretic separation is demonstrated.

Original languageEnglish (US)
Pages (from-to)329-337
Number of pages9
JournalJournal of Microcolumn Separations
Volume10
Issue number4
DOIs
StatePublished - Jan 1 1998

Keywords

  • Capillary electrophoresis
  • Membrane collection
  • Peroxyoxalate chemiluminescence
  • Postcolumn detection
  • Radionuclide detection

ASJC Scopus subject areas

  • Filtration and Separation
  • Mechanical Engineering

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