Determination of Fluorescence-Labeled Asparaginyl-Oligosaccharide in Glycoprotein by Reversed-Phase Ultraperformance Liquid Chromatography with Electrospray Ionization Time-of-Flight Mass Spectrometry.

Determination of Fluorescence-Labeled Asparaginyl-Oligosaccharide in Glycoprotein by Reversed-Phase Ultraperformance Liquid Chromatography with Electrospray Ionization Time-of-Flight Mass Spectrometry. Research Abstract Details 

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Determination of Fluorescence-Labeled Asparaginyl-Oligosaccharide in Glycoprotein by Reversed-Phase Ultraperformance Liquid Chromatography with Electrospray Ionization Time-of-Flight Mass Spectrometry. Abstract Text:

Takamasa Kurihara,Jun Zhe Min,Toshimasa Toyo'oka,Takeshi Fukushima,Shinsuke Inagaki,Takamasa Kurihara,Jun Zhe Min,Toshimasa Toyo'oka,Takeshi Fukushima,Shinsuke Inagaki,Takamasa Kurihara,Jun Zhe Min,Toshimasa Toyo'oka,Takeshi Fukushima,Shinsuke Inagaki,

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.

Determination of Fluorescence-Labeled Asparaginyl-Oligosaccharide in Glycoprotein by Reversed-Phase Ultraperformance Liquid Chromatography with Electrospray Ionization Time-of-Flight Mass Spectrometry. Publishing Authors By Initials

T Kurihara,JZ Min,T Toyo'oka,T Fukushima,S Inagaki,T Kurihara,JZ Min,T Toyo'oka,T Fukushima,S Inagaki,T Kurihara,JZ Min,T Toyo'oka,T Fukushima,S Inagaki,

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Determination of Fluorescence-Labeled Asparaginyl-Oligosaccharide in Glycoprotein by Reversed-Phase Ultraperformance Liquid Chromatography with Electrospray Ionization Time-of-Flight Mass Spectrometry. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Analytical chemistry

VOLUME: 79

Page Numbers: 8694-8

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ISSN: 0003-2700

DAY: 13

MONTH: 10

YEAR: 2007

Determination of Fluorescence-Labeled Asparaginyl-Oligosaccharide in Glycoprotein by Reversed-Phase Ultraperformance Liquid Chromatography with Electrospray Ionization Time-of-Flight Mass Spectrometry. Information

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LANGUAGE: eng

NlmUniqueID: 370536

Determination of Fluorescence-Labeled Asparaginyl-Oligosaccharide in Glycoprotein by Reversed-Phase Ultraperformance Liquid Chromatography with Electrospray Ionization Time-of-Flight Mass Spectrometry. Keywords Mesh Terms:

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Grant and Affiliation Information for Determination of Fluorescence-Labeled Asparaginyl-Oligosaccharide in Glycoprotein by Reversed-Phase Ultraperformance Liquid Chromatography with Electrospray Ionization Time-of-Flight Mass Spectrometry.

AFFILIATION: Division of Bio-Analytical Chemistry, School of Pharmaceutical Sciences, and Global COE Program, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.

Country: United States

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MEDLINETA: Anal Chem

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