Fluorescence detection of proteins and amino acids in capillary electrophoresis using a post-column sheath flow reactor

Paula G. Coble, Aaron T. Timperman

Research output: Contribution to journalArticlepeer-review

Abstract

A simple laser-induced fluorescence detection method for proteins and amino acids in capillary electrophoresis is reported. A sheath flow cell is utilized as a post-column reactor for fluorescence derivatization of proteins and amino acids by addition of o-phthaldialdehyde-2-mercaptoethanol to the sheath fluid. With the use of a 50 μm I.D. capillary, the limits of detection for carbonic anhydrase are 0.73 nM or 1.8 amol which represents a five- and two-fold improvement, respectively, over the best results previously reported for post-column detection. In addition, separation efficiencies up to 8.07.105 are achieved, and the detector response is linear over three-orders of magnitude. These results demonstrate that mixing is adequate and the reaction kinetics are rapid enough to provide sensitive detection with this approach. Also, because this post-column derivatization scheme requires no instrumental changes to a typical sheath flow cell detector, the system can be used for detection of pre-column labeled analytes and for native fluorescence detection. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)309-315
Number of pages7
JournalJournal of Chromatography A
Volume829
Issue number1-2
DOIs
StatePublished - Dec 1 1998
Externally publishedYes

Keywords

  • Amino acids
  • Detection, electrophoresis
  • Fluorescence detection
  • Proteins

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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