Simultaneous determination of N-acetylglucosamine, N-acetylgalactosamine, N-acetylglucosaminitol and N-acetylgalactosaminitol by gas-liquid chromatography.
A gas-liquid chromatographic procedure is described which will concomitantly separate and quantitate N-acetylglucosamine, N-acetylgalactosamine, N-acetylglucosaminitol and N-acetylgalactosaminitol in a single analytical run. The hexosamines, as their O-methyloximes, and the hexosaminitols can be separated as either their per-O-acetylated or per-O-trimethylsilylated derivatives. This method is particularly useful with samples that possess both N-acetylhexosaminitols and N-acetylhexosamines as are seen with N-linked oligosaccharides that are cleaved from glycoproteins by alkaline borohydride treatment. This procedure demonstrates a range of acceptable linearity of 1-1000 nmoles for each type of amino sugar.