Protocol for targeted analysis of oxylipins in rat plasma and mouse macrophage samples by high-resolution UHPLC-MS/MS

Oxylipins are bioactive oxygenated fatty acid derivatives that act as signaling mediators in diverse physiological and pathological processes. Here, we present a protocol for the simultaneous analysis of 125 oxylipins in rat plasma and mouse macrophage samples. We describe steps for preparing standards and calibration curves, extracting oxylipins, and performing ultra-high-performance liquid chromatography coupled to high-resolution tandem mass spectrometry (UHPLC-MS/MS). We also detail procedures for using staRoxy, an original R package for oxylipin abundance data analysis. For complete details on the use and execution of this protocol, please refer to Chaves-Filho et al.,¹ Kolmert et al.² and Yapici et al.³.