Methodologies and Procedures Employed in the Identification and Quantitation of Lipid Mediators via LC-MS/MS
Liquid chromatography tandem mass spectrometry (LC-MS/MS) is a sensitive and robust approach for the identification and quantitation of lipid mediators including the specialized pro-resolving mediators (resolvins, maresins, protectins and lipoxins), and the classic eicosanoids. Due to the relatively low concentration of these molecules in biological samples, this methodology is classically coupled with solid phase extraction (SPE) techniques that enhance the sensitivity of the assay. Herein, we provide a step-by-step description of instrumentation employed in the both SPE and LC-MS/MS and detailed methodologies employed in the extraction of these lipid mediators and their identification and quantitation using LC-MS/MS.
Solid-phase extraction coupled with liquid chromatography tandem mass spectrometry provides a robust and sensitive approach for the identification and quantitation of specialized pro-resolving mediators (lipoxins, resolvins, protectins, and maresins), their pathway markers and the classic eicosanoids. Here, we provide a detailed description of the methodologies employed for the extraction of these mediators from biological systems, setup of the instrumentation, sample processing, and then the procedures followed for their identification and quantitation.