Elucidation of post-translational modifications to proteins, such as glycosylations or phosphorylations, is one of the major issues concerning ongoing proteomic studies. To reduce general sample complexity, a necessary prerequisite is specific enrichment of peptide subsets prior to mass spectrometric sequencing. Regarding analysis of overall N-glycosylation sites in the past, this has been achieved by several approaches proving to be more or less complicated and specific. Here, we present a novel strategy to target N-glycosylation sites with application to platelet membrane proteins. Initial aqueous two phase partitioning for membrane enrichment and single step strong cation exchange based purification of glycopeptides resulted in identification of 148 glycosylation sites on 79 different protein species. While 69% of these sites were not annotated in the Swiss-Prot database before, a high number of 75% plasma membrane localized proteins was analyzed. Furthermore, miniaturizations as well as relative quantification are comprised in the developed method suggesting further use in other proteome projects. Results on platelet glycosylation sites may imply impact on research of bleeding disorders as well as potential new functions in inflammation and immunoactivity.