Larissa R Diniz, Adriano B Chaves-Filho, Pedro H F Jordão, Rosangela S Santos, Marina T Botana, Johan Kolmert, Ana C A Zucão, Marteinn T Snaebjornsson, Eduardo M Reis, Almut Schulze, Britta Brügger, Craig E Wheelock, Sayuri Miyamoto
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.