This chapter discusses the ω and (ω-1)-hydroxylation of eicosanoids and fatty acids by high-performance liquid chromatography (HPLC). Cytochromes P450 of the IVA family catalyze the hydroxylation of a number of endogenous substrates including prostaglandins (PG), leukotriene B4 (LTB4), and fatty acids at their ω and ω-1 carbon atoms. ω-Oxidation is a major route in prostaglandin metabolism, with a large proportion of prostaglandins being excreted in the urine as ω-oxidized products. Hydroxylated products are formed enzymatically by incubating fatty acids or prostaglandins with lung microsomes from pregnant rabbits, liver microsomes from diethylhexyl phthalate (DEHP)-treated rats, or purified cytochromes P450 from these tissues in reconstituted enzyme systems. The enzymatic rates of formation of products are calculated based on the percentage of radioactivity for the given metabolite relative to the total radioactivity eluting from the HPLC column.
ASJC Scopus subject areas
- Molecular Biology