An Advanced LC–MS/MS Platform for the Analysis of Specialized Pro-Resolving Lipid Mediators
- 565 Downloads
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.
KeywordsLC–MS Inflammation resolution Pro-resolving mediators Synovial fluid LTE4 EPA
The presented research was partially funded by the Prof. Jan Veltkamp Fonds of the LUMC. We thank Dr. Makoto Arita from the Graduate School of Pharmaceutical Sciences at the University of Tokyo for the generous supply of synthetic standard material of RvE1, RvE2, 18R-RvE3 and 18S-RvE3.
Conflict of interest
The authors declare no conflict of interest.
- 2.von Schacky C, Marcus AJ, Safier LB, Ullman HL, Islam N, Broekman MJ, Fischer S (1990) Platelet-neutrophil interactions. 12S,20- and 5S,12S-dihydroxyeicosapentaenoic acids: two novel neutrophil metabolites from platelet-derived 12S-hydroxyeicosapentaenoic acid. J Lipid Res 31(5):801–810Google Scholar
- 3.Isobe Y, Arita M, Matsueda S, Iwamoto R, Fujihara T, Nakanishi H, Taguchi R, Masuda K, Sasaki K, Urabe D, Inoue M, Arai H (2012) Identification and structure determination of novel anti-inflammatory mediator resolvin E3, 17,18-dihydroxyeicosapentaenoic acid. J Biol Chem 287(13):10525–10534. doi: 10.1074/jbc.M112.340612 CrossRefGoogle Scholar
- 5.Hsiao H-M, Sapinoro RE, Thatcher TH, Croasdell A, Levy EP, Fulton RA, Olsen KC, Pollock SJ, Serhan CN, Phipps RP, Sime PJ (2013) A novel anti-inflammatory and pro-resolving role for resolvin D1 in acute cigarette smoke-induced lung inflammation. PLoS ONE 8(3):e58258. doi: 10.1371/journal.pone.0058258 CrossRefGoogle Scholar
- 11.Giera M, Ioan-Facsinay A, Toes R, Gao F, Dalli J, Deelder AM, Serhan CN, Mayboroda OA (2012) Lipid and lipid mediator profiling of human synovial fluid in rheumatoid arthritis patients by means of LC–MS/MS. Biochim Biophys Acta 1821(11):1415–1424. doi: 10.1016/j.bbalip.2012.07.011 CrossRefGoogle Scholar
- 15.Strassburg K, Huijbrechts AL, Kortekaas K, Lindeman J, Pedersen T, Dane A, Berger R, Brenkman A, Hankemeier T, Duynhoven J, Kalkhoven E, Newman J, Vreeken R (2012) Quantitative profiling of oxylipins through comprehensive LC–MS/MS analysis: application in cardiac surgery. Anal Bioanal Chem 404(5):1413–1426. doi: 10.1007/s00216-012-6226-x CrossRefGoogle Scholar
- 18.Dalli J, Serhan CN (2012) Specific lipid mediator signatures of human phagocytes: microparticles stimulate macrophage efferocytosis and pro-resolving mediators. Blood 120(15):e60–e72. doi: 10.1182/blood-2012-04-423525