Ultraviolet native fluorescence detection in capillary electrophoresis using a metal vapor NeCu laser.
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.
Zhang
X
X
Department of Chemistry, University of Illinois, Urbana 61801, USA.
Sweedler
J V
JV
D000818:Animals; D004298:Dopamine / Q000032:analysis; D019075:Electrophoresis, Capillary / Q000379:methods / Q000592:standards; D005240:Feasibility Studies; D007834:Lasers / Q000592:standards; D008856:Microscopy, Fluorescence; D008974:Mollusca / Q000166:cytology; D009474:Neurons / Q000737:chemistry; D010455:Peptides / Q000032:analysis*; D012680:Sensitivity and Specificity; D012701:Serotonin / Q000032:analysis; D014466:Ultraviolet Rays
D010455:Peptides; D012701:Serotonin; D004298:Dopamine