1. HILIC/ESI-MS determination of gangliosides and other polar lipid classes in renal cell carcinoma and surrounding normal tissues
- Author
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Roman Hájek, Lukáš Opálka, Michal Holčapek, Miroslav Lísa, Robert Jirásko, David Vrána, Vladimír Študent, Kateřina Vávrová, Bohuslav Melichar, Eva Cífková, Maria A. Khalikova, and Marcel Matzenauer
- Subjects
0301 basic medicine ,Spectrometry, Mass, Electrospray Ionization ,Electrospray ionization ,Mass spectrometry ,01 natural sciences ,Biochemistry ,Analytical Chemistry ,03 medical and health sciences ,Gangliosides ,Lipidomics ,Pi ,medicine ,Humans ,Phosphatidylserines ,Carcinoma, Renal Cell ,Kidney ,Ganglioside ,Chromatography ,Chemistry ,Hydrophilic interaction chromatography ,010401 analytical chemistry ,Reference Standards ,0104 chemical sciences ,030104 developmental biology ,medicine.anatomical_structure ,lipids (amino acids, peptides, and proteins) ,Hydrophobic and Hydrophilic Interactions - Abstract
Negative-ion hydrophilic liquid chromatography-electrospray ionization mass spectrometry (HILIC/ESI-MS) method has been optimized for the quantitative analysis of ganglioside (GM3) and other polar lipid classes, such as sulfohexosylceramides (SulfoHexCer), sulfodihexosylceramides (SulfoHex2Cer), phosphatidylglycerols (PG), phosphatidylinositols (PI), lysophosphatidylinositols (LPI), and phosphatidylserines (PS). The method is fully validated for the quantitation of the studied lipids in kidney normal and tumor tissues of renal cell carcinoma (RCC) patients based on the lipid class separation and the coelution of lipid class internal standard with the species from the same lipid class. The raw data are semi-automatically processed using our software LipidQuant and statistically evaluated using multivariate data analysis (MDA) methods, which allows the complete differentiation of both groups with 100% specificity and sensitivity. In total, 21 GM3, 28 SulfoHexCer, 26 SulfoHex2Cer, 10 PG, 19 PI, 4 LPI, and 7 PS are determined in the aqueous phase of lipidomic extracts from kidney tumor tissue samples and surrounding normal tissue samples of 20 RCC patients. S-plots allow the identification of most upregulated (PI 40:5, PI 40:4, GM3 34:1, and GM3 42:2) and most downregulated (PI 32:0, PI 34:0, PS 36:4, and LPI 16:0) lipids, which are primarily responsible for the differentiation of tumor and normal groups. Another confirmation of most dysregulated lipids is performed by the calculation of fold changes together with T and p values to highlight their statistical significance. The comparison of HILIC/ESI-MS data and matrix-assisted laser desorption/ionization mass spectrometric imaging (MALDI-MSI) data confirms that lipid dysregulation patterns are similar for both methods. Graphical abstract ᅟ.
- Published
- 2018