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. 2014 Jun 17;79(1):139–149. doi: 10.1111/tpj.12547

Figure 1.

Figure 1

Mass spectrometric and thin-layer chromatographic analysis of Rosa glycosylinositol phosphorylceramides (GIPCs). (a) ESI-MS analysis of GIPC extract from Rosa cell culture. The spectrum was acquired in the negative ion mode. Abbreviations: Hex, hexose residue (probably α-mannose); HexA, hexuronic acid residue (probably α-glucuronic acid); Pent, pentose residue; Ins, myo-inositol; P, phosphate; Cer, phytoceramide. Inset: proposed structure of the predominant GIPC species; phytoceramide moiety in grey box. (b) ESI-MS/MS (collision-induced dissociation spectrum) analysis of the predominant Hex-HexA-Ins-P-Cer peak seen in (a) as the [M-2H]2− ion at m/z 630. Nitrogen was used as collision gas in a Q-TRAP instrument, with the collision energy set to −40 eV. The standard nomenclature for glycolipid fragmentation has been applied (Costello and Vath, 1990; Levery et al., 2001). Inset: proposed identity of the ion at m/z = 438.4, indicating an h24:0 ceramide moiety. (c, d) Thin-layer chromatography (TLC) of GIPC extract. Lipids were chromatographed in CHCl3/CH3OH/4 m NH4OH (9:7:2, by vol.) with 0.2 m ammonium acetate (Kaul and Lester, 1978) and located by orcinol reagent (c) or periodic acid–Schiff staining (d). Lipid bands are labelled: 1, Hex-HexA-Ins-P-Cer; 2, (Hex)2-HexA-Ins-P-Cer; 3, Pent-(Hex)2-HexA-Ins-P-Cer.