Sample preparation for the analysis of complex carbohydrates by multicapillary gel electrophoresis with light-emitting diode induced fluorescence detection.
This paper evaluates various sample preparation methods for multicapillary gel electrophoresis based glycan analysis to support electrokinetic injection. First the removal of excess derivatization reagent is discussed. Although the Sephadex G10 filled multiscreen 96-well filter plate and Sephadex G10 filled pipet tips enabled increased analysis sensitivity, polyamide DPA-6S pipet tips worked particularly well. In this latter case an automated liquid handling system was used to increase purification throughput, necessary to feed the multicapillary electrophoresis unit. Problems associated with the high glucose content of such biological samples as normal human plasma were solved by applying ultrafiltration. Finally, a volatile buffer system was developed for exoglycosidase-based carbohydrate analysis.
Olajos
Marcell
M
Horvath Laboratory of Bioseparation Sciences, Institute of Analytical Chemistry and Radiochemistry, University of Innsbruck, Innsbruck, Austria.
Hajós
Péter
P
Bonn
Guenther K
GK
Guttman
András
A
D000818:Animals; D002021:Buffers; D002417:Cattle; D004566:Electrodes; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D005453:Fluorescence; D006026:Glycoside Hydrolases / Q000737:chemistry; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D009005:Monosaccharides / Q000032:analysis; D011134:Polysaccharides / Q000032:analysis*; D011721:Pyrenes / Q000737:chemistry; D013194:Staining and Labeling; D014835:Volatilization
C098210:1-aminopyrene-3,6,8-trisulfonic acid; D002021:Buffers; D007202:Indicators and Reagents; D009005:Monosaccharides; D011134:Polysaccharides; D011721:Pyrenes; D006026:Glycoside Hydrolases