Structural investigation of oxidized arabinoxylan oligosaccharides by negative ionization HILIC-qToF-MS

Owing to the strong structure-function relationship of polysaccharides, the targeted modification of polysaccharides is attracting widespread interest in various fields, such as food industry, nutritional science, and biomedical research. Apart from intended functionalization, polysaccharide degradation mediated by hydroxyl radicals (HO˙) occurs in various industrial processes such as food processing. In particular, the oxidative degradation of feruloylated arabinoxylan (AX), a linearly-branched polysaccharide in cereals, causes chain scissions, and introduces new functional groups in the fiber, which can potentially modify the physicochemical properties and the functionalities of AX. However, the precise characterization of those structural modifications remains challenging due to the diversity of the oxidation products formed, the high molecular weight, and the relatively low quantity of newly formed functional groups. In this paper, selective (TEMPO-mediated) and random (Fenton) oxidations of several commercial xylo- and arabinoxylan oligosaccharides (A)XOS were studied as model systems by hydrophilic interaction UPLC-MS2 in negative ion resolution mode to identify potential oxidation products. An in-depth identification of acidic (A)XOS oxidation products derived from TEMPO-mediated oxidation provided novel insights in the selective functionalization of isomeric oligosaccharides. Furthermore, MS2 enabled the precise localisation of both glycosidic linkages and functional groups in oxidized (A)XOS. An innovative combination of an enzymatic sample preparation combined with a subsequent HILIC-MS2 analysis enabled the unprecedented comprehensive characterization of Fenton-induced oxidation products derived from AX. In future, this holistic analytical approach will enable the characterization of both selective and non-selective AX oxidation procedures in various applications.