Differential mobility separation of leukotrienes and protectins.
Differential mobility spectrometry (DMS) is capable of separating stereoisomeric molecular ions based on their mobility in an oscillating electrical field with an asymmetric waveform. Thus, it is an "orthogonal" technique to chromatography and (tandem) mass spectrometry. Bioactive lipids, particularly of the eicosanoid and docosanoid class feature numerous stereoisomers, which exhibit a highly specific structure-activity relationship. Moreover, the geometry of these compounds also reflects their biochemical origin. Therefore, the unambiguous characterization of related isomers of the eicosanoid and docosanoid classes is of fundamental importance to the understanding of their origin and function in many biological processes. Here we show, that SelexION DMS technology coupled to μLC-MS/MS is capable of differentiating at least five closely related leukotrienes partially coeluting and (almost) unresolvable using LC-MS/MS only. We applied the developed method to the separation of LTB4 and its coeluting isomer 5S,12S-diHETE in murine peritoneal exudate cells, showing that LTB4 is present only after zymosan A injection while its isomer 5S,12S-diHETE is produced after saline (PBS) administration. Additionally, we show that the SelexION technology can also be applied to the separation of PD1 and PDX (10S,17S-diHDHA), two isomeric protectins.
†Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2300RC Leiden, The Netherlands.|‡Department of Rheumatology, Leiden University Medical Center, Albinusdreef 2, 2300RC Leiden, The Netherlands.
Hulda S
HS
Jónasdóttir
§SCIEX Germany GmbH, Landwehrstrasse 54, 64293 Darmstadt, Germany.
Cyrus
C
Papan
§SCIEX Germany GmbH, Landwehrstrasse 54, 64293 Darmstadt, Germany.
Sebastian
S
Fabritz
∥Institut des Biomolécules Max Mousseron (IBMM), UMR 5247-CNRS, University of Montpellier, 34090 Montpellier, France.
Laurence
L
Balas
∥Institut des Biomolécules Max Mousseron (IBMM), UMR 5247-CNRS, University of Montpellier, 34090 Montpellier, France.
Thierry
T
Durand
⊥Faculty of Medicine, Biomedical Center, School of Health Sciences, University of Iceland, Vatnsmyrarvegi 16, 101 Reykjavik, Iceland.
Ingibjorg
I
Hardardottir
⊥Faculty of Medicine, Biomedical Center, School of Health Sciences, University of Iceland, Vatnsmyrarvegi 16, 101 Reykjavik, Iceland.|#Department of Immunology and Center for Rheumatology Research, Landspitali-The National University of Iceland, 101 Reykjavik, Iceland.
Jona
J
Freysdottir
†Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2300RC Leiden, The Netherlands.
Martin
M
Giera
D000818:Animals; D018957:CD59 Antigens / Q000737:chemistry / Q000302:isolation & purification*; D002853:Chromatography, Liquid; D007536:Isomerism; D015289:Leukotrienes / Q000737:chemistry / Q000302:isolation & purification*; D051379:Mice; D013057:Spectrum Analysis / Q000379:methods*; D053719:Tandem Mass Spectrometry
D018957:CD59 Antigens; D015289:Leukotrienes