Open access
Protocol for targeted analysis of oxylipins in rat plasma and mouse macrophage samples by high-resolution UHPLC-MS/MS
L. Diniz
Adriano B. Chaves-Filho
P. H. F. Jordão
Rosangela S. Santos
Marina T. Botana
J. Kolmert
A. C. A. Zucão
M. Snaebjornsson
E. M. Reis
Almut Schulze
B. Brügger
C. Wheelock
Sayuri Miyamoto
Medicine
Abstract
Summary 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