Developing a Bioanalytical Toolbox for Human Milk Oligosaccharide Characterization

Human milk oligosaccharides are one of the most challenging classes of molecules to characterize, largely owing to their high degree of isomeric heterogeneity. Thus, new analytical methodologies are required to improve the confidence of their characterization. Herein, the use of cyclic ion mobility separations coupled with mass spectrometry (cIMS-MS) is presented as a new analytical tool for human milk oligosaccharide analyses. By combining two dimensional cIMS-MS information, stemming from both high-resolution collision cross section values and isotopic shifts, we have developed a workflow for better resolving key human milk oligosaccharide isomers. The use of tandem mass spectrometry as well as tandem ion mobility spectrometry will also be discussed.