Capillary liquid chromatography-mass spectrometry using graphitised carbon stationary phase and ion trap mass spectrometry was shown to be a powerful technique for analysing glycosaminoglycans digested with endoglycosidases. Commonly found disaccharides from heparin/heparan sulphate digests at sub nanomole levels were found to be separated by mass and/or retention time and detected by negative ion electrospray mass spectrometry predominantly as [M - H]- ions using a standard electrospray interface and flow rate between 6-10 μL/min. Graphitised carbon liquid chromatography-fragmentation mass spectrometry provided sequence data of disaccharides and oligosaccharides. Sequence information was obtained from either collision of the [M - H] - ions (low sulphated disaccharides) or of the [M + Na - 2H] - ions (highly sulphated disaccharides). This separation and identification method of endoglycosidase digestion and sample preparation using a combination of cation exchange and graphitised carbon, was used to successfully analyse digests of keratan sulphate (keratanase) and heparin (heparinase) standards, and hyaluronic acid (hyaluronidase) from synovial fluid samples.
|Number of pages||9|
|Journal||Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences|
|Publication status||Published - 25 Sep 2005|
- Heparan sulphate
- Hyaluronic acid
- Keratan sulphate