Details

Autor(en) / Beteiligte

• Lísa, Miroslav
• Řehulková, Helena
• Hančová, Eliška
• Řehulka, Pavel

Titel

Lipidomic analysis using hydrophilic interaction liquid chromatography microgradient fractionation of total lipid extracts

Ist Teil von

• Journal of Chromatography A, 2021-09, Vol.1653, p.462380, Article 462380

Ort / Verlag

Elsevier B.V

Erscheinungsjahr

2021

Quelle

Access via ScienceDirect (Elsevier)

Beschreibungen/Notizen

• •Lipidomic analysis using a simple microgradient LC device.•Lipid class fractionation of total lipid extracts.•Fractionation using a HILIC microcolumn and a gradient in a microsyringe.•Comprehensive RP-LC analysis of collected lipid classes. Lipidomic samples are complex mixtures of structurally different species of a wide range of concentrations providing challenges in their characterization. In this work, we present a proof of concept for the application of a simple microgradient liquid chromatography device on the detailed analysis of lipid classes. Our lipidomic analysis is based on a lipid class microgradient fractionation of a total lipid extract using an in-house-prepared hydrophilic interaction liquid chromatography microcolumn followed by RP-LC/MS of the collected lipid class fractions. The final fractionation method uses a 40-mm-long microcolumn of 500 μm ID with silica stationary phase obtained from a commercially available chromatographic column and the microgradient of the mobile phase prepared in a microsyringe using methyl tert-butyl ether (MTBE) – methanol – water – ammonium acetate mixtures of various elution strengths. MTBE total lipid extract is directly separated by microgradient elution into lipid classes according to their polarity, which enables the collection of isolated fractions of most lipid classes. The method has been applied to the fractionation of porcine brain extract into nonpolar lipids, hexosylceramides, phosphoethanolamines, phosphocholines, sphingomyelins, and lysophosphocholines classes. Achieved repeatability, recovery, and advanced lipid coverage prove the applicability of the microgradient fractionation of total lipid extract for the comprehensive lipidomic analysis.