Ultraviolet native fluorescence detection in capillary electrophoresis using a metal vapor NeCu laser

Anal Chem. 2001 Nov 15;73(22):5620-4. doi: 10.1021/ac010458z.

Abstract

Ultraviolet (UV) excitation for laser-induced native fluorescence (LINF) detection in capillary electrophoresis (CE) offers impressive performance figures of merit when assaying peptides containing tyrosine or tryptophan residues, catecholamines, indolamines, and a number of other classes of analytes with appreciable fluorescence when excited by UV radiation. One of the largest drawbacks of native fluorescence detection schemes in CE-LINF systems has been the expense and the complexity of the lasers required for excitation in the deep UV wavelength range of 200-300 nm. An improved "turnkey" NeCu laser operating at 248.6 nm interfaced to a sheath flow-based CE system obtains a performance similar to that of large frame frequency-doubled Ar ion lasers. The detection limits for serotonin and dopamine (27 nM and 8 microM, respectively, for approximately 3-nL injection) are similar to those obtained using a frequency-doubled Ar ion laser at 257 nm (21 nM and 8 microM, respectively). An example of the detection of serotonin-related analytes from a single-cell electropherogram demonstrates the performance of such a system for mass-limited measurements.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Dopamine / analysis
  • Electrophoresis, Capillary / methods
  • Electrophoresis, Capillary / standards
  • Feasibility Studies
  • Lasers / standards
  • Microscopy, Fluorescence
  • Mollusca / cytology
  • Neurons / chemistry
  • Peptides / analysis*
  • Sensitivity and Specificity
  • Serotonin / analysis
  • Ultraviolet Rays

Substances

  • Peptides
  • Serotonin
  • Dopamine