Analysis of N‐acetylaminosugars by CE: A comparative derivatization study

N‐linked or O‐linked glycans derived from glycoprotein processing carry, an N‐acetylglucosamine or an N‐acetylgalactosamine respectively, at their reducing termini. The presence of the N‐acetylamino group on C‐2 of reducing sugar residues has been reported to hamper the derivatization reaction with a chromophore at the anomeric centre. In this paper N‐acetyllactosamine, N‐acetylglucosamine, N‐acetylgalactosamine and several other neutral monosaccharides are coupled to three different dyes (4‐aminobenzonitrile, 2‐aminopyridine, 2‐aminobenzoic acid (2‐AA)) by reductive amination and analysed by CE with UV detection. The 2‐AA derivatives showed the lowest concentration detection limits, varying approximately in the 2–3 μM range for the saccharides tested including the N‐acetamido ones. The possibility to separate and detect with the same sensitivity ten 2‐AA‐labelled monosaccharides mainly found in mammalian or plant glycoproteins in a single CE run is highlighted. The analysis has been carried out in less than 25 min using the borate‐complexation method in CZE mode. The influence of the strength of the acid used as catalyst in the chemical modification of the sugars with 2‐AA is also shortly addressed.