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. Author manuscript; available in PMC: 2020 Nov 1.
Published in final edited form as: Biochim Biophys Acta Gen Subj. 2019 Aug 6;1863(11):129409. doi: 10.1016/j.bbagen.2019.08.002

Figure 3. RP-HPLC of anionic N-glycans of male honeybee larvae.

Figure 3

PNGase Ar/F-released N-glycans were subject to solid phase extraction, whereby the anionic-enriched fraction was eluted with 40% acetonitrile/0.1% trifluoroacetic acid, prior to fluorescent labelling and chromatography on either an RP-amide column (A; at pH 4) or a Kinetex XB-C18 column (B; at pH 6); each fraction was collected and subject to MALDI-TOF MS with m/z values in bold italics for [M-H]- ions of sulphated structures. The annotations in the Symbolic Nomenclature for Glycans are based on elution time, MS/MS and digestion data (see Figures 4-6 and Supplementary Figures 5-8). The columns were calibrated in terms of glucose units. Note that on both columns sulphation (S) and antennal fucosylation lead to large shifts to lower retention time, whereas this effect is only seen on the RP-amide column for phosphoethanolamine (PE)-modified structures (highlighted in blue boxes); structures with two different β1,2/β1,4-antennae, which are a special feature of larvae, are highlighted in purple boxes.