Globotriaosylceramide-related biomarkers of fabry disease identified in plasma by high-performance thin-layer chromatography - densitometry- mass spectrometry.

摘要: Identification of 19 molecular species globotriaosylceramides (Gb3) in extracts from a Fabry's plasma patient and healthy control was performed by High-Performance Thin-Layer Chromatography (HPTLC)-densitometry online coupling to Mass Spectrometry (MS). Separation carried out on LiChrospher plates using Automated Multiple Development (AMD). Densitometry twin combining detection the visible at 550 nm, through previous on-plate orcinol derivatization, Ultraviolet 190 non-impregnated plate. The latter directly coupled an ion-trap mass spectrometer automated elution-based interface. Gb3 species, which were identified HPTLC- Electrospray (+)-MS confirmed MS/MS or HPTLC-Atmospheric Pressure Chemical Ionization (+)-MS, are: five isoforms saturated Gb3; seven methylated with two additional double bonds. Twelve these previously reported as biomarkers lysosomal disorder Liquid Chromatography-MS-based method, other are structurally similar, closely related them. Saturated migrated plate one separated peaks corresponding migration zone ceramide trihexosides standard. Instead, unsaturated co-migrated sphingomyelin species. Ion intensity ESI-MS profiles show that higher concentration than sample. Before applying (TLC)-MS interface HPTLC peaks, its positioning precision first studied tri-hexosides model compound. This provided information peak broadening splitting during migration.