A novel condition for capillary electrophoretic analysis of reductively aminated saccharides without removal of excess reagents

Electrophoresis
Masahiro YodoshiShigeo Suzuki

Abstract

We have identified novel CE conditions for the separation of 7-amino-4-methylcoumarin-labeled monosaccharides and oligosaccharides from glycoproteins. Using a neutrally coated capillary and alkaline borate buffer containing hydroxypropylcellulose and ACN, saccharide derivatives form anionic borate complexes, which move from the cathode to the anode in an electric field and are detected near the anodic end. Excess labeling reagents and other fluorescent products remain at the cathodic end. Fluorimetric detection using an LED as a light source enables determination of monosaccharide derivatives with good linearity between at least 0.4 and 400 μM, may correspond to 140 amol to 140 fmol. The lower LOD (S/N = 5) is only 80 nM in the sample solution (ca. 28 amol). The results were comparable to reported values using fluorometric detection LC. The method was also applied to the analysis of oligosaccharides that were enzymatically released from glycoproteins. Fine resolution enables profiling of glycans in glycoproteins. The applicability of the method was examined by applying it to other derivatives labeled with nonacidic tags such as ethyl p-aminobenzoate- and 2-aminoacridone-labeled saccharides.

References

Dec 16, 1998·Electrophoresis·S Suzuki, S Honda
Dec 16, 1998·Electrophoresis·F T Chen, R A Evangelista
Feb 5, 2010·Biological Chemistry·Dieter VanderschaegheNico Callewaert
Dec 15, 2010·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·David J Harvey
Apr 12, 2011·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Masahiro YodoshiShigeo Suzuki

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Citations

Feb 17, 2015·Proteomics. Clinical Applications·Edward S X MohNicolle H Packer
Jul 28, 2016·Journal of Pharmaceutical and Biomedical Analysis·Sachio YamamotoShigeo Suzuki
Mar 13, 2018·Chemical Reviews·Grace LuLisa A Holland

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