Separation of 8-aminonapthalene-1,3,6-trisulfonic acid-labelled neutral and sialylated N-linked complex oligosaccharides by capillary electrophoresis

摘要: Abstract Complex oligosaccharides, both neutral and sialylated, were derivatized with 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS) separated by capillary electrophoresis. The derivatization reaction was carried out in a total volume of 2 μl. peaks detected laser induced fluorescence detection using the 325-nm line HeCd laser. Concentration mass limits 5·10 −8 M 500 amol, respectively, could be achieved. limiting step for higher sensitivity is not detector performance, however, but chemistry limit 2.5·10 −6 . Two labelling protocols established, one overnight at 40°C other 2.5-h time 80°C. Neutral oligosaccharides labelled either protocol. However, sialylated hydrolysed when Low nanomole to picomole amounts oligomannose-type complex-type oligosaccharide mixtures less than 8 min excellent resolution phosphate background electrolyte pH 2.5. linear relationship between electrophoretic mobility charge-to-mass ratios ANTS conjugates used peak assignment. Further, influence three-dimensional structure complex on their migration behaviour discussed. suitability subsequent separation analysis patterns demonstrated libraries derived from ovalbumin bovine fetuin. For assignment are compared those oligomannose mixtures. efficiency 120 000 theoretical plates times 10 superior state-of-the-art chromatographic methods electrophoresis methods. A difference 0.06 found sufficient baseline oligosaccharides.