Separation of Phospholipids Derived from Biological Extracts Using a Solid Core Reversed Phase HPLC Column
semanticscholar(2012)
Abstract
Introduction The role of membrane lipids on cell surfaces is known to be of key importance to cell function and inter-cellular communication. Interest in lipidomics (the large-scale study of pathways and networks of cellular lipids in biological systems1) is growing rapidly, and understanding which lipids are present within cells is an important aspect of biological studies. The separation, detection and classification of lipids by conventional LC-MS methods can be a challenging application. Lipids comprise a range of different classes and generally include a polar head group together with at least one attached hydrocarbon chain (Figure 1). Their analysis is complicated by the wide variation in composition and structures present in any biological extract combined with high retention on C18 reversed phases. Solid-core stationary phases have been shown to be able to yield UHPLC-levels of resolution due to the minimized resistance to mass transfer by the diffusional path of analytes being limited by the depth of the porous outer layer. The optimized packing results in more uniform paths through the LC column. These can be attained without the unwanted side effect of elevated backpressure which is associated with smaller particle sizes. In this application note a separation protocol for the analysis of lipids by LC-MS using a solid-core stationary phase is presented.
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