Size exclusion chromatography/mass spectrometry applied to the analysis of polysaccharides.
A novel method for analysing polysaccharide materials is described which employs size-exclusion chromatography (SEC) followed by detection by on-line electrospray ionisation mass spectrometry (ESI-MS) and off-line matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS). It is demonstrated through SEC/ESI ion trap mass spectrometry that the formation of multiply charged oligomer ions, which bind up to five sodium cations, allows the rapid analysis of polysaccharide ions with molecular weights in excess of 9 kDa. MALDI spectra generated from fractionation of the effluent collected from the same SEC separation are shown to be in good agreement with the ESI spectra with respect to molecular weight distributions and types of ions generated. ESI and MALDI mass spectra of samples obtained from sequential graded ethanol precipitation and SEC fractionation of acid and enzymatically digested arabinoxylan polysaccharides show important structural differences between polysaccharide fragments. In addition, a comparison is made between the mass spectra of native and permethylated SEC-separated fragments of acid and enzymatically treated arabinogalactan. Linkage information of the permethylated arabinogalactan oligomers can be rapidly established through the use of on-line SEC/ESI-MS( n) experiments.
Deery
M J
MJ
DuPont (UK) Ltd, Cereals Innovation Centre, Block B, The Mill Site, 40 Station Road, Cambridge CB1 2UJ, UK. md289@mole.bio.cam.ac.uk
Stimson
E
E
Chappell
C G
CG
D001426:Bacterial Proteins; D002240:Carbohydrate Sequence; D003911:Dextrans; D005685:Galactans / Q000032:analysis / Q000737:chemistry; D006868:Hydrolysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000032:analysis / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry; D012015:Reference Standards; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods* / Q000592:standards; D043325:Xylan Endo-1,3-beta-Xylosidase; D014990:Xylans / Q000032:analysis / Q000737:chemistry; D014995:Xylosidases; D001616:beta-Galactosidase
D001426:Bacterial Proteins; D003911:Dextrans; D005685:Galactans; D009844:Oligosaccharides; D011134:Polysaccharides; D014990:Xylans; C085118:arabinoxylan; D014995:Xylosidases; D001616:beta-Galactosidase; D043325:Xylan Endo-1,3-beta-Xylosidase; C109490:galactanase A; C005653:arabinogalactan