A rapid and highly sensitive microchip electrophoresis of mono- and mucin-type oligosaccharides labeled with 7-amino-4-methylcoumarin
| Autor: | Yuki Asada, Shigeo Suzuki, Sachio Yamamoto, Mitsuhiro Kinoshita, Eriko Nagai |
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| Rok vydání: | 2014 |
| Předmět: |
Detection limit
chemistry.chemical_classification Chromatography Resolution (mass spectrometry) Chemistry Oligosaccharides Sensitivity and Specificity Biochemistry Analytical Chemistry Anode Electrophoresis Microchip Electrophoresis Capillary electrophoresis Coumarins Reagent Monosaccharide Quantitation Range Glycoproteins |
| Zdroj: | Analytical and Bioanalytical Chemistry. 407:1499-1503 |
| ISSN: | 1618-2650 1618-2642 |
| Popis: | A selective separation method using a poly(methylmethacrylate) microchip was developed for 7-amino-4-methylcoumarin-labeled saccharides in a crude reaction mixture. In an alkaline borate buffer, saccharide derivatives formed strong anionic borate complexes. These complexes moved from the cathode to the anode in an electric field and were detected near the anode. Excess labeling reagents and other foreign substances remained at the inlet reservoir. A confocal fluorimetric detection system enabled the determination of monosaccharide derivatives with good linearity between at least 5 and 100 nM, corresponding to 50 fmol to 1 pmol per injection. The lower limit of detection (signal-to-noise = 5) was 2 nM. The sensitivity and linear quantitation range were comparable to reported values using fluorometric detection, capillary electrophoresis, or liquid chromatography. Application of the method showed excellent resolution in the analysis of O-linked glycans chemically released from glycoproteins. |
| Databáze: | OpenAIRE |
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