An Advanced LC-MS/MS Platform for the Analysis of Specialized Pro-Resolving Lipid Mediators

Here we present an advanced platform for the analysis of specialized pro-resolving mediators (SPM), including a set of pro-inflammatory lipids and respective biochemical pathway markers. The platform is characterized by a largely simplified sample preparation protocol, employing methanol protein-precipitation instead of solid phase extraction. Sample preparation and analysis can be carried out in 96-well format, ensuring higher throughput. In addition, faster analysis times and higher sensitivities have been achieved when compared to other platforms. In total 36 lipid mediators-SPM, their pathway markers and a set of pro-inflammatory lipids including three internal standards-are analyzed within an 11 min LC-MS/MS run. The method was validated using human plasma, including repeatability, linearity, recovery, matrix effects, and accuracy. The presented platform was applied in the analysis of synovial fluid post-mortem samples and cellular extracts from polymorphonuclear cells as well as whole blood treated with ionophore in the presence of the polyunsaturated fatty acid eicosapentaenoic acid.