Deep UV laser-induced fluorescence detection of unlabeled drugs and proteins in microchip electrophoresis.
Deep UV fluorescence detection at 266-nm excitation wavelength has been realized for sensitive detection in microchip electrophoresis. For this purpose, an epifluorescence setup was developed enabling the coupling of a deep UV laser into a commercial fluorescence microscope. Deep UV laser excitation utilizing a frequency quadrupled pulsed laser operating at 266 nm shows an impressive performance for native fluorescence detection of various compounds in fused-silica microfluidic devices. Aromatic low molecular weight compounds such as serotonin, propranolol, a diol, and tryptophan could be detected at low-micromolar concentrations. Deep UV fluorescence detection was also successfully employed for the detection of unlabeled basic proteins. For this purpose, fused-silica chips dynamically coated with hydroxypropylmethyl cellulose were employed to suppress analyte adsorption. Utilizing fused-silica chips permanently coated with poly(vinyl alcohol), it was also possible to separate and detect egg white chicken proteins. These data show that deep UV fluorescence detection significantly widens the application range of fluorescence detection in chip-based analysis techniques.
Schulze
Philipp
P
Abteilung für Chromatographie, Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470 Mülheim an der Ruhr, Germany.
Ludwig
Martin
M
Kohler
Frank
F
Belder
Detlev
D
D002919:Chymotrypsinogen / Q000032:analysis; D004527:Egg Proteins / Q000032:analysis; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D007834:Lasers; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D009113:Muramidase / Q000032:analysis; D004364:Pharmaceutical Preparations / Q000032:analysis*; D011433:Propranolol / Q000032:analysis; D011409:Propylene Glycols / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D012701:Serotonin / Q000032:analysis; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods; D014362:Trypsinogen / Q000032:analysis; D014364:Tryptophan / Q000032:analysis; D014466:Ultraviolet Rays
D004527:Egg Proteins; D004364:Pharmaceutical Preparations; D011409:Propylene Glycols; D011506:Proteins; D012701:Serotonin; D014364:Tryptophan; D014362:Trypsinogen; D002919:Chymotrypsinogen; D011433:Propranolol; D009113:Muramidase