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Glycosylation is an important post-translational modification of proteins. Many diseases, such as cancer, have proved to be related to aberrant glycosylation. High throughput quantitative methods have gained attention recently in the study of glycomics. With the development of high-resolution mass spectrometry, the sensitivity of detection in glycomics has largely improved; however, most of the commonly used MS-based techniques are focused on relative quantitative analysis, which can hardly provide direct comparative glycomic quantitation results. In this study, we developed a novel multiplex glycomic analysis method on an LC–ESI–MS platform. Reduced glycans were stable isotopic labeled during the permethylation procedure, with the use of iodomethane reagents CH₂DI, CHD₂I, CD₃I, ¹³CH₃I, ¹³CH₂DI, ¹³CHD₂I, ¹³CD₃I, and CH₃I. Up to 8-plex glycomic profiling was possible in a single analysis by LC …