Premium
A novel condition for capillary electrophoretic analysis of reductively aminated saccharides without removal of excess reagents
Author(s) -
Yodoshi Masahiro,
Ikeda Natsumi,
Yamaguchi Naoko,
Nagata Mana,
Nishida Noriaki,
Kakehi Kazuaki,
Hayakawa Takao,
Suzuki Shigeo
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200612
Subject(s) - chemistry , chromatography , monosaccharide , reagent , capillary electrophoresis , electrophoresis , fluorescence , glycoprotein , organic chemistry , biochemistry , physics , quantum mechanics
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.