Verspreet J, Holmgaard Hansen A, Dornez E, Courtin CM, Harrison SJ. Anal Bioanal Chem. 2014 May 14. In this paper, a new liquid chromatography-mass spectrometry (LC-MS) method for the analysis of complex fructan mixtures is presented. In this method, columns with a trifunctional C18 alkyl stationary phase (T3) were used and their performance compared with that of a porous graphitized carbon (PGC) column. The separation of fructan isomers with the T3 phase improved clearly in comparison with the PGC phase, and retention times were lower and more stable. When the T3-based method was applied on a wheat grain extract, multiple fructan isomers could be discerned, even for fructans with a degree of polymerization of 10. This indicates that wheat grain fructans do not, or not only, have a simple linear structure. The presented method paves the way for elucidation of fructan structures in complex mixtures that contain many structural isomers.