Determination of fluorescence-labeled asparaginyl-oligosaccharide in glycoprotein by reversed-phase ultraperformance liquid chromatography with electrospray ionization time-of-flight mass spectrometry
| Autor: | Shinsuke Inagaki, Toshimasa Toyo'oka, Jun Zhe Min, Takeshi Fukushima, Takamasa Kurihara |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Detection limit
Spectrometry Mass Electrospray Ionization Chromatography Molecular Structure Chemistry Ovalbumin Electrospray ionization Ion chromatography Fluorescence spectrometry Oligosaccharides Reversed-phase chromatography High-performance liquid chromatography Sensitivity and Specificity Fluorescence spectroscopy Analytical Chemistry Animals Time-of-flight mass spectrometry Asparagine Chickens Hydrophobic and Hydrophilic Interactions Chromatography Liquid Fluorescent Dyes Glycoproteins |
| Zdroj: | Analytical chemistry. 79(22) |
| ISSN: | 0003-2700 |
| Popis: | Eight fluorescence reagents, i.e., DBD-F, NBD-F, DNS-Cl, NDA, PSC, FITC, Fmoc-Cl, and DMEQ-COCl, which are reactive to an amino functional group, were tested for the labeling of asparaginyl-oligosaccharides in a glycoprotein. Although the optimal reaction conditions and the fluorescence maximal wavelengths were different for each reagent, the highly sensitive fluorescence detection at the femtomole level of Disialo-Asn (a representative asparaginyl-oligosaccharide) was obtained from the labeling utilizing these reagents. Among them, PSC was the most reliable reagent in terms of detection sensitivity (approximately 3 fmol, signal-to-noise ratio of 5 (S/N = 5) on the chromatogram). However, the structural information could not be obtained from the fluorescence detection. Thus, the on-line determination of a real sample was carried out by UPLC-ESI-TOF-MS. The detection limit of the PSC-labeled Disialo-Asn by selected-ion chromatography was 58 fmol (S/N = 5). When the proposed procedure was applied to the determination of oligosaccharides in ovalbumin, 15 species of PSC-labeled oligosaccharides possessing Man, GlcNAc, and Gal units were identified from the UPLC-ESI-TOF-MS. The number of identified oligosaccharides was relatively greater than the method using Fmoc-Cl. Based on the ovalbumin results, the proposed labeling with PSC followed by UPLC-ESI-TOF-MS detection seems to be useful for the on-line asparaginyl-oligosaccharide analysis. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|