Fig. 1: 4D-glycosphingolipidomics using RP-LC-TIMS-PASEF MS for glycosphingolipids analysis. | Nature Communications

Fig. 1: 4D-glycosphingolipidomics using RP-LC-TIMS-PASEF MS for glycosphingolipids analysis.

From: Extended coverage of human serum glycosphingolipidome by 4D-RP-LC TIMS-PASEF unravels association with Parkinson’s disease

Fig. 1

a Symbol Nomenclature for Glycan13,14 (https://www.ncbi.nlm.nih.gov/glycans/snfg.html) used throughout figures: Glucose (blue circle), Galactose (yellow circle), N-acetylglucosamine (blue square), N-acetylgalactosamine (yellow square), N-acetylneuraminic acid (purple diamond), and Fucose (red triangle); b Base peak chromatogram showing separation of glycosphingolipids (GSLs) in customized standard mixture using reversed-phase liquid chromatography. A customized standard mixture was prepared from commercial standards of GM4, GM3, GM2, GM1, GD3, GD2, GD1a, GD1b, GT1b, and GQ1 species in an equimolar concentration of 5 pmol/µL, with 20 µL injected onto the column. GSLs were separated based on their ceramide moiety and degree of sialylation; c Partial separation of GD1a 18:1;O2/18:0 and GD1b 18:1;O2/18:0 in extracted ion chromatogram (EIC) and in extracted ion mobilogram (EIM) of m/z 917.48 in negative ion mode and of m/z 919.50 in positive ion mode. The PASEF spectra obtained from the distinct regions of the partially separated extracted ion chromatogram and mobilogram in negative ion mode demonstrate the differential pattern of the diagnostic fragment ions at nominal m/z 581 characteristic for GD1b species, at higher abundance at CCS value of 456.4 Å2. In positive ion mode, the di-sialo element was exclusively found in the spectrum corresponding to the mobilogram at CCS value of 473.5  Å2. In both spectra of GD1a and GD1b in positive ion mode diagnostic fragment ions of the 18:1;O2 sphingoid base and the accompanying neutral loss of fatty acyl from the ceramide moiety ions are underscoring the ceramide structure; d Three-dimension (m/z, collisional cross section (CCS), retention time (RT)) distribution of GSLs in customized standard mixture (average value of each descriptor from n = 3 measurements) (Supplementary Data 1); e Two-dimension (m/z, CCS) distribution of GSLs in customized standard mixture (average value of each descriptor from n = 3 measurements). Source data are provided as a Source Data file.

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