Figure 2. Experimental workflow of lipid mediators analysis using MRM^HR.

Various biological samples, such as lung tissue, plasma and culture medium, were prepared by solid phase extraction (SPE) for subsequent UHPLC-MS/MS analysis. The scheme of MRM^HR analysis was represented by 5-HETE detection (precursor ion at m/z 319.2 and fragment ions at m/z 257.2279, 203.1805, 115.0395 and 59.0151) in a complex mixture. The lipids were identified and quantified using bioinformatics software package. A library of HR-MS/MS profile was created for each metabolite quantified. A partial validation of MRM^HR method was conducted according FDA recommendations.