An Advanced LC–MS/MS Platform for the Analysis of Specialized Pro-Resolving Lipid Mediators
article
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. © 2014, Springer-Verlag Berlin Heidelberg.
Topics
EPAInflammation resolutionLC–MSLTE4Pro-resolving mediatorsCell extractIcosapentaenoic acidIonophoreLipidMethanolPolyunsaturated fatty acidSpecialized pro resolving lipidUnclassified drugIsolation and purificationLipid analysisLiquid chromatographyMass spectrometryPolymorphonuclear cellSolid phase extractionSynovial fluidValidation study
TNO Identifier
524069
ISSN
00095893
Source
Chromatographia, 78(5-6), pp. 391-401.
Pages
391-401
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