Direct electrospray-ionization mass spectrometric analysis of the major ganglioside from crucian carp liver after thin layer chromatography

• Published in: Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 1999, Vol.122 (1), p.83-88
• Main Authors: Hildebrandt, Herbert, Jonas, Ute, Ohashi, Masako, Klaiber, Iris, Rahmann, Hinrich
• Format: Article
• Language: English
• Subjects: Animals; Birds; Carps - physiology; Chromatography, Thin Layer - methods; Collision induced dissociation tandem mass spectrometry; Cricetinae; Electrospray ionization mass spectrometry; Fish; Ganglioside analysis; Gangliosides - chemistry; Glycosphingolipid structure; Liver - chemistry; Mass Spectrometry - methods; Muscles - chemistry; Oreochromis mossambicus; Thin layer chromatography; Tissue Distribution; Vertebrate liver
• ISSN: 1096-4959; 0305-0491; 1879-1107
• DOI: 10.1016/S0305-0491(98)10152-9

Ganglioside patterns from crucian carp brain, muscle, and liver as well as liver gangliosides of roach, carp, the cichlid Oreochromis mossambicus, pigeon, dwarf hamster, and calf were comparatively analyzed by high performance thin layer chromatography (HPTLC). To achieve a rapid estimation on potentially interesting ganglioside compounds, electrospray-ionization mass spectrometry (MS) was directly applied to a chloroform/methanol extract of the major TLC band of crucian carp liver. The spectrum, obtained from a few micrograms of this crude biological sample, revealed a series of peaks corresponding to GM4-like monosialoganglioside species. GC-MS analysis revealed hydroxylated fatty acids ranging from 2 h 20 min:0 to 2 h 26 min:0 for the [M'H] − ions of m/ z 1061–1145. Collision induced dissociation tandem MS/MS of the major peak with a [M'H] − ion of m/ z 1117 demonstrated the presence of N-acetylneuraminic acid as sialic acid compound. The sugar composition was confirmed by GLC as galactose and sialic acid in a 1:1 molar ratio. Thus, the structure of the ion at m/ z 1117 is N-acetylneuraminylgalactosylceramide (NeuAc-Gal-Cer) with the long chain base d18:1 and the hydroxylated fatty acid 2 h 24 min:0. The results demonstrate for the first time unambiguously that NeuAc-Gal-Cer is the main ganglioside fraction in fish liver and that electrospray ionization-mass spectrometry (ESI-MS) can be used to elucidate the chemical composition of a ganglioside fraction obtained by convenient extraction of a HPTLC band.