Electrospray tandem mass spectrometry of underivatised acetylated xylo-oligosaccharides

Acetylated neutral (XylnAcm) and acidic xylo‐oligosaccharides (XylnAcmMeGlcA, and XylnAcmMeGlcAHex) obtained by partial acid hydrolysis of Eucalyptus globulus wood glucuronoxylans and fractionated by preparative ligand exchange/size‐exclusion chromatography were identified by electrospray ionisation mass spectrometry (ESI‐MS). Low molecular weight acetylated xylo‐oligosaccharides were studied by ESI‐tandem mass spectrometry (MS/MS). All the acetylated xylo‐oligosaccharides showed an abundant ion due to the neutral loss of 60 Da (CH3CO2H) in the MS/MS spectra. The presence of diacetylated xylo‐oligosaccharides was confirmed by the ions formed by loss of two molecules of acetic acid. Furthermore, characteristic [XylresAc2+Na]+ and [XylAc2+Na]+ ions, and ions due to loss of XylAc2, indicate that both acetyl groups are located in the same Xyl residue. On the other hand, losses of XylresAc and XylAc are also observed as well as [XylresAc+Na]+ and [XylAc+Na]+ , indicating the location of both acetyl groups in different Xyl residues, in some cases even in adjacent xyloses. The MS/MS spectra of triacetylated xylo‐oligosaccharides were complex due to the presence of different isobaric xylo‐oligosaccharides containing the acetyl groups at different locations in the xylo‐oligosaccharide backbone. In the MS/MS spectra of acidic xylo‐oligosaccharides, the ion at m/z 387, [XylresAcMeGlcA+Na]+, indicates that the acetyl groups are preferentially linked to Xyl substituted with MeGlcA. However, acidic xylo‐oligosaccharides with the acetyl and 4‐O‐methylglucuronic acid groups in different Xyl residues were also identified. In neutral and in acidic xylo‐oligosaccharides several possible locations of the acetyl groups were identified, namely at terminal positions. In summary, ESI‐MS/MS is shown to be a powerful tool for the characterisation of acetylated patterns in complex mixtures of oligosaccharides. Copyright © 2005 John Wiley & Sons, Ltd.