Abstract
Raffinose family oligosaccharides (RFOs) are commonly applied as functional food ingredients. RFOs mixture derived from Eupatorium were analyzed by hydrophilic interaction liquid chromatography-evaporative light scattering detector (HILIC-ELSD) method with a Click Xion column, and then separated by graphitized carbon black (GCB) chromatography. RFOs with a specific degree of polymerization (DP) from 3 to 6 were obtained. The specific ions from glycosidic bond cleavage and cross-ring cleavage of RFOs were produced in Electrospray-collision-induced dissociation-mass spectroscopy/mass spectroscopy (ESI-CID-MS/MS), which are in accordance with the linkage of (1→6) between galactans. The anomeric configuration and glycosidic linkage of RFOs were confirmed to be [α-Gal-(1→6)]n-α-Glc-(1→2)- β-Fru (n = 1–4) by 2D-NMR. In this study, we developed a HILIC-ELSD method for the compositional analysis of RFOs and obtained RFOs with a specific DP and high purity, and their structures were completely elucidated. Our work provides a basis for further research on the structure–activity relationship of RFOs.
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