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Glycoscreening by on‐line sheathless capillary electrophoresis/electrospray ionization‐quadrupole time of flight‐tandem mass spectrometry
Author(s) -
Zamfir Alina,
PeterKatalinic Jasna
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200107)22:12<2448::aid-elps2448>3.0.co;2-a
Subject(s) - capillary electrophoresis , chemistry , electrospray ionization , mass spectrometry , chromatography , electrospray , tandem mass spectrometry , quadrupole , quadrupole time of flight , capillary electrophoresis–mass spectrometry , analytical chemistry (journal) , capillary action , materials science , physics , atomic physics , composite material
An analytical approach based on sheathless on‐line coupling of capillary electrophoresis (CE) and electrospray ionization (ESI) quadrupole time‐of‐flight (QTOF) mass spectrometry (MS) has been developed for providing new insight into the characterization of carbohydrate mixtures. The home‐built sheathless CE/ESI microspray interface consisting of an one‐piece CE column has been optimized and applied for the screening of a complex mixture of O ‐glycosylated peptides obtained and previously purified from urine of patients suffering from N ‐acetylhexosaminidase deficiency. The separation efficiency and the resolution obtained in CE/UV experiments could be reproduced with high sensitivity in on‐line CE/MS runs under mild ES‐Inegative ion mode conditions, due to the compatibility of the reconsidered CE‐QTOF/MS operating parameters, microspray tip performance and the use of MS friendly volatile CE buffer system as ammonium acetate/ammonium hydroxide. By employing the high speed automated “on‐the‐fly” MS‐MS/MS switching abilities of QTOF/MS instrument, the multicomponent sample could be separated and subsequently submitted to both MS and low‐energy collision‐induced dissociation of selected precursor ions. Using the on‐line CE‐ESI‐MS/MS coupling, glycosylated sialylated peptides undetectable in complex mixtures by a direct ESI/MS analysis could be identified.