Lipids are a diverse array of biomolecules involved in numerous biochemical and physiological processes. Their immense structural diversity has brought challenges to lipidomics, specifically in determining the unique molecular species among thousands of different isoforms. Lipids tend to have long fatty acyl chains, which enhance the difficulty of analyzing them with mass spectrometry. Nevertheless, mass spectrometry and the electrospray ionization remain the most reliable methods for generating quantitative and qualitative data. In order to produce a successful analysis, the lipid samples must be carefully isolated and purified, so as to minimize the generation of closely related molecules when these are ionized. An important part of the isolation and purification of lipids is the extraction procedure. This step removes the unwanted and potentially interfering materials. The most widely used methods are the Folch method and the Blight & Dyer method, which employ a chloroform/methanol system. Despite the fact that these methods work efficiently, the use of chloroform, a known carcinogen, is not recommended. Moreover, the lipids are contained in the lower phase of the extract, meaning that the upper phase must be penetrated, thus, causing sample contamination. The aim of this research is to develop and validate a method that overcomes the drawbacks of the aforesaid methods. The improved method includes a three-step extraction with methanol, and the upper phase contains the pure lipid extract. The purified lipids are then coupled to high resolution mass spectrometry (MS) using a heated electrospray ionization (HESI) source in negative mode, given that negative ions have higher abundance for most lipids.

Improved chloroform-free lipid extraction method on long-chain lipids followed by ultra-high performance liquid chromatography-high resolution mass spectrometry (uhplc-hrms)

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.