0000000000828573
AUTHOR
Estelle Sibille
Sample preparation and analysis of gangliosides and phospholipids from plasma by liquid chromatography coupled to mass spectrometry
Gangliosides (GG) and phospholipids (PL) are two classes of complex and polar lipids exhibiting a wide structural heterogeneity. LC/MS offers an access to this huge diversity but also faces some challenges. Efficient extraction and purification sample methods are thus required to perform accurate and reliable analyses. To this end, GG and PL were extracted from human plasma with three solvent mixtures (CHCl3/MeOH/H2O) of increasing polarity, in comparison with the reference Folch method. GG were further purified with a Phree Phospholipid Removal column, whose sorbent is designed to selectively remove PL. GG and PL classes were then separated under HILIC conditions before analysis using a Qq…
Gangliosides in the retina: a comparative inventory with brain and other ocular structures
International audience
Apprehending ganglioside diversity: a comprehensive methodological approach
Gangliosides make a wide family of glycosphingolipids ubiquitously expressed in mammalian tissues and particularly abundant in the brain and nervous system. They exhibit a huge diversity due to structural variations in both their oligosaccharidic chain and ceramide moiety, which represent a real analytical challenge. Since their discovery in the 1940s, methods have persistently improved until the emergence of Liquid Chromatography/Mass Spectrometry (LC/MS) which offers a high level of specificity and sensitivity and is suitable with high-throughput profiling studies. We describe here a comprehensive approach relying on various techniques and aiming at fully characterizing gangliosides in bi…
Gangliosides characterization by high resolution mass spectrometry and ion mobility
Gangliosides (GG) are sialic acid-containing glycosphingolipids particularly abundant in the nervous system. They exhibit a wide variety of structures depending on both the oligosaccharide chain and the ceramide moiety. About 200 GG species have been described so far, but their diversity and biological roles are far from being completely elucidated. An efficient method of identification and quantification is crucial to apprehend this huge heterogeneity. In this study, we focused on the characterization of the ceramide portion of GG, identifying the long chain base (LCB) and the fatty acid. A commercial standard of GD3 was analysed with both a Thermo LTQ-Orbitrap XLTM and a Waters Synapt G2-…
4812 - A0028 Gangliosides in the retina: a comparative inventory with brain and other ocular structures
Purpose: Gangliosides (GG) make an extremely diverse family of glycosphingolipids particularly abundant in the brain and neural tissues. While it is known that alterations in GG metabolism are associated with visual defects, the precise biological roles of these compounds in the retina are still largely unknown. In this context, we performed an exhaustive inventory and thorough characterization of the human eye GG aiming to reveal specificities and common features of the retina compared to other ocular tissues and brain.Methods: Retina, RPE/Choroid, ciliary body, optic nerve and brain samples were collected from deceased human donors (N=7). GG were extracted and analysed by a comprehensive …