Abstract
This chapter discusses the advances in the chromatography of lipids. The method of preparation of a lipid extract constitutes one of the most critical parts of lipid analysis by chromatographic methods. Extraction with organic solvents provides a simple method of isolating all major phospholipids regardless of their fatty-acid composition. The isolation of the minor acidic phosphoinositol phosphatides (PtdInsPs) requires acidification, which causes the destruction of plasmalogens unless they are removed first. Good laboratory practice recommends that the lipid extractions be performed in the presence of butylated hydroxy toluene (BHT) or other antioxidants, especially when working with small amounts of lipid. Because PtdInsPs and their products of hydrolysis are degraded very rapidly, it is essential either to extract the samples immediately after collection or to freeze the samples in situ after collection and store them at −70oC. In situ fixation of samples by freeze clamping or by microwave irradiation may be helpful in arresting the degradation of phosphate esters.
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