期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1203, 期 2, 页码 137-145出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2008.07.053
关键词
fluorescent probe; 7-amino-4-methylcoumarin; glycoprotein; negative-ion electrospray mass spectrometry
资金
- Ministry of Education, Culture, Sports, Science and Technology (MEXT), 2007-2011
Reductive amination reaction using 7-anlino-4-niethylcoumarin (AMC) as a fluorescent probe enabled analyses of glycoproteins' monosaccharides and N-linked oligosaccharides. Reductive amination of N-acetylhexosamines and AMC using sodium cyanoborohydride or dimethylamine-borane complex indicated slight recovery of derivatives, but pyridine-borane achieved better recoveries. Reversed-phase high-performance liquid chromatography (HPLC) analyses of monosaccharides constituting glycoprotein glycans using fluorimetric detection revealed linearity for 0.2 fmol to 1 pmnol, with less than 5% RSD quantitation reproducibility. Reversed-phase HPLC analyses of glycoprotein glycans, combined with negative-ion electrospray ionization mass spectrometry (LC-ESI-MS), enabled their Structural determination. Using this highly hydrophobic reagent, AMC-labeled oligosaccharides displayed one-order to two-order higher ESI-MS intensity than derivatives labeled using other reagents. (C) 2008 Elsevier B.V. All rights reserved.
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