Conformational separation of monosaccharides of glycoproteins labeled with 2-aminoacrydone using microchip electrophoresis

The conformational separation of monosaccharides labeled with fluorescent 2‐aminoacrydone (AMAC) was performed by electrophoresis on a plastic microchip with light‐emitting diode confocal fluorescence detection. The AMAC‐labeled five neutral monosaccharide mixture (D‐glucose (Glc), D‐mannose, D‐galactose, L‐fucose, and D‐xylose) or two amino monosaccharide mixture (N‐acetyl‐D‐glucosamine and N‐acetyl‐D‐galactosamine) were well separated at pH 8.5 and 0.5% w/v methylcellulose of 200 mM borate buffer conditions using microchip electrophoresis. The separation was successfully performed considering the difference in stability of the complex between the hydroxyl residue of the monosaccharide and borate ions, and we found that 200 mM and pH 8.5 of borate buffer conditions were critical. High‐speed separation for the neutral monosaccharides (50 s) and for amino monosaccharides (70 s) was attained at a 400 V/cm of electric field condition, showing all peak resolutions were greater than 0.9% and RSD of mobility were less than 1.9%. The detection limits of 0.86 µM for Glc and