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HILIC-HPLC/ESI-MS analysis of gangliosides in biological samples
Autoři: Hájek Roman | Jirásko Robert | Lísa Miroslav | Cífková Eva | Holčapek Michal
Rok: 2017
Druh publikace: ostatní - přednáška nebo poster
Strana od-do: nestránkováno
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Jazyk Název Abstrakt Klíčová slova
eng HILIC-HPLC/ESI-MS analysis of gangliosides in biological samples Gangliosides are acid glycosphingolipids containing one or more sialic acids (N˗acetylneuraminic acid, NeuAc) in the oligosaccharide chain. These lipids are abundant in neural tissues and play important roles in the nervous system and many physiological processes in cells, such as memory control, cell signaling, neuronal recovery, neuronal protection or apoptosis. Gangliosides are also related to some serious diseases such as cancer, Alzheimer disease, Tay-Sachs disease, and Fabry disease. Recently, gangliosides have also become an attractive target of biomarker discovery. Gangliosides are extracted from tissues and body fluids by chloroform, methanol and water extraction according to Folch method with minor modifications. Aqueous phase containing gangliosides is further purified on C18 solid-phase extraction column to remove undesired salts before LC˗MS. The analytical method for the analysis of gangliosides was developed using hydrophilic interaction liquid chromatography coupled to negative-ion electrospray ionization mass spectrometry(HILIC/ESI-MS). The optimization of chromatographic parameters including the buffer type, its concentration, and pH value was performed to achieve the optimal HILIC separation with regard to the best sensitivity and chromatographic resolution of studied compounds. The optimized method using the porous shell particle column Ascentis Si (15x2.1 mm, 3 μm) pH 6.1 adjusted by acetic acid and the gradient of acetonitrile and 10 mM of aqueous ammonium acetate for the lipidomic analysis of human kidney, lungs, plasma, urine, erythrocytes, and porcine brain. 142 gangliosides from 19 classes are identified in biological samples, but the gangliosides are majority of detected only in the brain tissue. Gangliosides; HILIC; ESI-MS

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