title: Combined HILIC-ELSD/ESI-MS(n) enables the separation, identification and quantification of sugar beet pectin derived oligomers.
abstract: The combined action of endo-polygalacturonase (endo-PGII), pectin lyase (PL), pectin methyl esterase (fungal PME) and RG-I degrading enzymes enabled the extended degradation of methylesterified and acetylated sugar beet pectins (SBPs). The released oligomers were separated, identified and quantified using hydrophilic interaction liquid chromatography (HILIC) with online electrospray ionization ion trap mass spectrometry (ESI-IT-MS(n)) and evaporative light scattering detection (ELSD). By MS(n), the structures of galacturonic acid (GalA) oligomers having an acetyl group in the O-2 and/or O-3 positions eluting from the HILIC column were elucidated. The presence of methylesterified and/or acetylated galacturonic acid units within an oligomer reduced the interaction with the HILIC column significantly compared to the unsubstituted GalA oligomers. The HILIC column enables a good separation of most oligomers present in the digest. The use of ELSD to quantify oligogalacturonides was validated using pure GalA standards and the signal was found to be independent of the chemical structure of the oligomer being detected. The combination of chromatographic and enzymatic strategies enables to distinguish SBPs having different methylesters and acetyl group distribution.
authors.affiliation: Wageningen University, Laboratory of Food Chemistry, P.O. Box 8129, 6700 EV Wageningen, The Netherlands.
authors.forename: C
authors.initials: C
authors.lastname: Remoroza
authors.affiliation: University of Münster, Department of Plant Biology and Biotechnology, Hindenburgplatz 55, 48143 Münster, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Cord-Landwehr
authors.affiliation: Wageningen University, Laboratory of Food Chemistry, P.O. Box 8129, 6700 EV Wageningen, The Netherlands.
authors.forename: A G M
authors.initials: AG
authors.lastname: Leijdekkers
authors.affiliation: University of Münster, Department of Plant Biology and Biotechnology, Hindenburgplatz 55, 48143 Münster, Germany.
authors.forename: B M
authors.initials: BM
authors.lastname: Moerschbacher
authors.affiliation: Wageningen University, Laboratory of Food Chemistry, P.O. Box 8129, 6700 EV Wageningen, The Netherlands.
authors.forename: H A
authors.initials: HA
authors.lastname: Schols
authors.affiliation: Wageningen University, Laboratory of Food Chemistry, P.O. Box 8129, 6700 EV Wageningen, The Netherlands.
authors.forename: H
authors.initials: H
authors.lastname: Gruppen
mesh_terms: D001230:Aspergillus / Q000201:enzymology; D027461:Beta vulgaris / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002853:Chromatography, Liquid; D002912:Chrysosporium / Q000201:enzymology; D006868:Hydrolysis; D057927:Hydrophobic and Hydrophilic Interactions; D008969:Molecular Sequence Data; D010368:Pectins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011096:Polygalacturonase / Q000378:metabolism; D011133:Polysaccharide-Lyases / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D010368:Pectins; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase; D011096:Polygalacturonase; D011133:Polysaccharide-Lyases; C027349:pectin lyase