Fatty acid metabolites, such as eicosanoids and docosanoids, play important biological roles and are strictly regulated by diverse enzymes. Here, we present two approaches for purifying their metabolites, including the phospholipid mediator platelet-activating factor (PAF), for quantification using liquid chromatography-mass spectrometry (LC-MS). We describe steps for tissue collection, lipid extraction, and lipid mediator purification. We then detail procedures for data analysis. This protocol allows the selection of a suitable technique for analyzing target molecules. For complete details on the use and execution of this protocol, please refer to Yamamoto et al. 1 • Instructions for the detection of fatty acid metabolites • Steps for the purification and enrichment of fatty acid metabolites • Selection of the purification procedure for the target metabolites Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Fatty acid metabolites, such as eicosanoids and docosanoids, play important biological roles and are strictly regulated by diverse enzymes. Here, we present two approaches for purifying their metabolites, including the phospholipid mediator platelet-activating factor (PAF), for quantification using liquid chromatography-mass spectrometry (LC-MS). We describe steps for tissue collection, lipid extraction, and lipid mediator purification. We then detail procedures for data analysis. This protocol allows the selection of a suitable technique for analyzing target molecules.
Hashidate-Yoshida et al. (Sun,) studied this question.