Separation and characterization of arachidonate-containing phosphoglycerides

The chapter presents a study on separation and characterization of arachidonate-containing phosphoglycerides. This chapter focuses on high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC) methods used in laboratory to achieve separation of arachidonate-containing phosphoglyceride classes, subclasses, and molecular species. Arachidonic acid metabolism is initiated by the mobilization of esterified arachidonate from cellular phosphoglycerides by the activation of intracellular phospholipases. There are a diverse group of phosphoglyceride molecular species that contain arachidonate in mammalian cells. The sn-1 position linkage of archidonate-containing phosphoglycerides is an important component that determines the biochemical pathways available to these molecules. In preparing the cells, human neutrophils are prepared from venous blood obtained from healthy human donors immediately before the neutrophil isolation. The chapter discusses HPLC separation of glycerolipid classes, TLC separation of choline- and ethanolamine-linked subclasses, HPLC separation of arachidonate-containing phosphoglyceride molecular species, and several related concepts. The chapter mentions the possibility to resolve a number of the major arachidonate-containing phosphoglycerides by using a modified reversed-phase system. This HPLC system has a major advantage over systems for diradylglyceride derivatives in that it requires much less sample preparation prior to HPLC.