Fluorescence detection system for capillary separations utilizing a liquid core waveguide with an optical fibre-coupled compact spectrometer.
A fluorescence detection system for capillary liquid separation methods is described. The system is based on a silica capillary coated with a low refractive index fluoropolymer Teflon AF that serves both as a separation channel and as a liquid core waveguide (LCW). A fibre-coupled laser excites separated analytes in a detection point and arising fluorescence is collected at one end of the LCW capillary into the other optical fibre which brings it to a compact charge-coupled device (CCD) array spectrometer installed in a desktop computer. No additional components such as focusing optics or filters are necessary. This system was used for detecting isoelectrically focused fluorescent low-molecular-mass pI (isoelectric point) markers and fluorescein isothiocyanate (FITC) labelled proteins. The ability of the system to acquire fluorescent spectra is also demonstrated.
Kostal
Vratislav
V
Department of Analytical Chemistry, Faculty of Science, Palacky University, tr. Svobody 26, 771 46 Olomouc, Czech Republic. kostal@iach.cz
Zeisbergerova
Marta
M
Slais
Karel
K
Kahle
Vladislav
V
D002138:Calibration; D019075:Electrophoresis, Capillary / Q000379:methods*; D005336:Fiber Optic Technology; D016650:Fluorescein-5-isothiocyanate; D007525:Isoelectric Focusing / Q000379:methods*; D007526:Isoelectric Point; D008904:Miniaturization; D011138:Polytetrafluoroethylene; D011506:Proteins / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D011506:Proteins; D011138:Polytetrafluoroethylene; D016650:Fluorescein-5-isothiocyanate