Protocol to purify fatty acid metabolites with two approaches for quantification using liquid chromatography-mass spectrometry

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.