The profound biological relevance of protein and lipid glycosylation has made glycomics (i.e., the comprehensive study of all glycans in a cell or organism), an indispensable field of research in the life sciences. Consequently, numerous strategies have been developed for a high-throughput analysis of complex glycan mixtures, with mass spectrometry (MS) playing a key role. In particular, nanoelectrospray ionization (ESI-) MS( n ), employing multiple cycles of isolation and fragmentation of native or derivatized precursor ions, is recognized as a highly valuable tool in this context, as it allows, at least in part, structural characterization of glycans without prior fractionation. This chapter describes suitable work flows for this purpose and illustrates both advantages and limitations for this type of analysis. Furthermore, the use of newly developed software tools for data handling is outlined.