Analysis of nonderivatized neutral and sialylated oligosaccharides by electrospray mass spectrometry.

An HPLC/MS method has been developed that allows rapid, direct analysis of underivatized sialylated as well as neutral oligosaccharides. The method involves the separation of oligosaccharides from salts and proteins using RP-HPLC with a formic acid/acetonitrile/water mobile phase system and on-line electrospray mass spectrometry analysis in the positive ion mode. Under the solution conditions employed, both neutral and acidic (sialylated) oligosaccharides are protonated and therefore detected. In contrast to MALDI-TOF MS, no loss of sialic acid is observed when operating in the positive ion mode. Furthermore, the capability of this method to provide quantitative estimates of the relative abundance of each oligosaccharide mass has been demonstrated using fetuin as a model compound.