Chiral chromatography–tandem mass spectrometry applied to the determination of pro-resolving lipid mediators

Homann, Julia; Lehmann, Christian; Kahnt, Astrid S.; Steinhilber, Dieter; Parnham, Michael J.; Geißlinger, Gerd; Ferreiròs, Nerea

doi:10.1016/j.chroma.2014.07.068

Cited by 15 publications

(14 citation statements)

References 40 publications

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“…We had to perform analytical runs on two different chiral phases in order to resolve distinct subsets, comprising most of the targeted monoepoxides on Lux Cellulose-3 and all monohydroxides as well as several mid-chain monoepoxides on Lux Amylose-1. Other investigators used Chiralpak AD-RH, Lux Amylose-2, or Lux Amylose-1 with 250 × 4.6 mm size to run chiral-LC coupled with ESI-MS/MS for the targeted lipidomics of resolvins and other specialized pro-resolving lipid mediators (8)(9)(10). Relatively long equilibration times needed before and in between successive analytical runs (see Technical notes and conditions for the analysis of biological samples in the Results section) are another problem concerning the moderate throughput in our chiral approach.…”

Section: Discussionmentioning

confidence: 99%

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Chiral lipidomics of monoepoxy and monohydroxy metabolites derived from long-chain polyunsaturated fatty acids

Blum

Doğan

Karber

et al. 2019

Journal of Lipid Research

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chidonic acid (AA), EPA, and DHA. Oxygenated PUFAs have also been termed "oxylipins" and comprise metabolites formed by cyclooxygenases (COXs), lipoxygenases (LOXs), and cytochrome P450 (CYP) enzymes as well as nonenzymatic oxidation reactions (1)(2)(3)(4). Current methods of targeted lipidomics allow high-throughput, comprehensive, and highly sensitive quantitation of oxylipins in biological and clinical samples (5). Most of these analytical approaches rely on LC coupled with MS/MS. Thereby, LC is performed on achiral stationary phases under reversedphase conditions and MS mostly uses ESI for efficient ionization of the analytes. These advanced methods can measure more than one hundred different oxylipin species in one analytical run, but are unable to distinguish between the enantiomers (5).The lack of enantiomeric resolution is an inherent property of the achiral-LC-ESI-MS/MS approaches. This feature limits the conclusions that can be drawn regarding the enzymatic versus nonenzymatic origin of oxylipin species or the biological significance of changes in the endogenous oxylipin profile, e.g., in the course of cardiovascular and inflammatory diseases. To address these questions, different strategies of targeted chiral lipidomics are under investigation (6). Chiral-LC has been primarily developed for normal-phase conditions using apolar solvent systems that preclude ESI application for efficient ionization. A way out of this problem is provided by electron capture atmospheric Abstract A chiral lipidomics approach was established for comprehensive profiling of regio-and stereoisomeric monoepoxy and monohydroxy metabolites of long-chain PUFAs as generated enzymatically by cytochromes P450 (CYPs), lipoxygenases (LOXs), and cyclooxygenases (COXs) and, in part, also unspecific oxidations. The method relies on reversedphase chiral-LC coupled with ESI/MS/MS. Applications revealed partially opposing enantioselectivities of soluble and microsomal epoxide hydrolases (mEHs). Ablation of the soluble epoxide hydrolase (sEH) gene resulted in specific alterations in the enantiomeric composition of endogenous monoepoxy metabolites. For example, the (R,S)/(S,R)-ratio of circulating 14,15-EET changed from 2.1:1 in WT to 9.7:1 in the sEH-KO mice. Studies with liver microsomes suggested that CYP/mEH interactions play a primary role in determining the enantiomeric composition of monoepoxy metabolites during their generation and release from the ER. Analysis of human plasma showed significant enantiomeric excess with several monoepoxy metabolites. Monohydroxy metabolites were generally present as racemates; however, Ca 2+ -ionophore stimulation of whole blood samples resulted in enantioselective increases of LOX-derived metabolites (12S-HETE and 17S-hydroxydocosahexaenoic acid) and COX-derived metabolites (11R-HETE). Our chiral approach may provide novel opportunities for investigating the role of bioactive lipid mediators that generally exert their physiological functions in a highly regio-and stereospecific manner.-Blum, M., I. Dog...

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Section: Discussionmentioning

confidence: 99%

“…This approach allows performing reversed-phase chiral-LC coupled with ESI-MS/MS detection and quantitation of the resolved stereoisomers. Reversed-phase chiral-LC-ESI-MS/MS has been successfully used for the determination of enantiomers as well as diastereomeric epimers of pro-resolving lipid mediators (8)(9)(10).…”

mentioning

confidence: 99%

Chiral lipidomics of monoepoxy and monohydroxy metabolites derived from long-chain polyunsaturated fatty acids

Blum

Doğan

Karber

et al. 2019

Journal of Lipid Research

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“…Because the detailed molecular mechanisms of SPM biosynthesis remain elusive, we established in vitro systems for both pathways to study basic principles. In addition, a detection method via LC-MS/MS was established (19). Representative standard chromatograms for the SPMs detected are depicted in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Quantitation of the analytes was performed by Analyst 1.5 (AB Sciex). A detailed description of the method used for the detection of SPM is available elsewhere (19).…”

Section: Extraction and Detection Of Lipid Mediators Via Lc-ms/msmentioning

confidence: 99%

Lipoxin and resolvin biosynthesis is dependent on 5‐lipoxygenase activating protein

et al. 2015

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Resolution of acute inflammation is an active process coordinated by proresolving lipid mediators (SPMs) such as lipoxins (LXs) and resolvins (Rvs), which are formed by the concerted action of 2 lipoxygenases (LOs). Because the exact molecular mechanisms of SPM biosynthesis are not completely understood, we aimed to investigate LX and D-type Rv formation in human leukocytes and HEK293T cells overexpressing leukotriene (LT) pathway enzymes. Activity assays in precursor (15-hydroxyeicosatetraenoic acids, 17-HDoHE)-treated granulocytes [polymorphonuclear leukocytes (PMNLs)] showed a strict dependence of LXA 4 /RvD 1 biosynthesis on cell integrity, and incubation with recombinant human 5-LO did not lead to LX or Rv formation. Pharmacologic inhibition of 5-LO activating protein (FLAP) by MK-886 inhibited LXA 4 /RvD 1 biosynthesis in precursor-treated PMNLs (drug concentration causing 50% inhibition ∼0.3/0.2 mM), as did knockdown of the enzyme in MM6 cells, and precursor-treated HEK293T overexpressing 5-LO produced high amounts of LXA 4 only in the presence of FLAP. In addition, inhibition of cytosolic phospholipase A 2a (cPLA 2a ) interfered with LXA 4 / RvD 1 formation from exogenous precursors in PMNLs. Furthermore, inhibition of the LT synthases LTA 4 hydrolase and LTC 4 synthase in PMNL/platelet coincubations augmented LXA 4 levels. These findings show that several enzymes known to be involved in the biosynthesis of proinflammatory LTs, such as FLAP and cPLA 2a , also contribute to LX and Rv formation.-

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“…Of the latter, only the derivative with the hydroxyl group in S-configuration is important for the raise of maresins and it could be confirmed by comparison with standards, that 14(S)-HDoHe is actually built as intermediary pathway marker. Using chiral chromatography, analytes differing in their configuration and, more importantly, isomers can be easily distinguished through enhanced interactions between the stationary phase and the analytes [16,17].…”

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confidence: 99%

Simultaneous determination of PUFA-derived pro-resolving metabolites and pathway markers using chiral chromatography and tandem mass spectrometry

Toewe

Balas

Geißlinger

et al. 2018

Analytica Chimica Acta

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Lipid mediators play an important role as biological messengers involved in inflammatory processes. Deriving from different polyunsaturated fatty acids, endogenously built mediators featuring both pro- and anti-inflammatory properties as well as pro-resolving lipid mediators and their biological precursors have been investigated. A newly developed method using chiral chromatography-tandem mass spectrometry on human plasma has demonstrated its suitability for the simultaneous determination of prostaglandins, lipoxins, D-series derived resolvins as well as protectins, maresin 1, leukotriene B4 and several precursors of them in order to yield information about metabolic pathways. Due to the matrix complexity, a solid phase extraction method using an octadecyl-modified silica gel cartridge was carried out. The developed method allows the determination of 34 analytes in 25 min showing enough selectivity as well as precision and accuracy (≤15% relative standard deviation, ≤15% relative error) in the calibration range of 0.1-10 ng mL or 0.2-20 ng mL depending on the analytes. Stability of the analytes in plasma has been demonstrated for at least 3 h at room temperature, 72 h in the autosampler and 60 days in the freezer at -80 °C. This method has been validated and shown its suitability for the determination of all studied analytes in human plasma samples.

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Chiral chromatography–tandem mass spectrometry applied to the determination of pro-resolving lipid mediators

Cited by 15 publications

References 40 publications

Chiral lipidomics of monoepoxy and monohydroxy metabolites derived from long-chain polyunsaturated fatty acids

Chiral lipidomics of monoepoxy and monohydroxy metabolites derived from long-chain polyunsaturated fatty acids

Lipoxin and resolvin biosynthesis is dependent on 5‐lipoxygenase activating protein

Simultaneous determination of PUFA-derived pro-resolving metabolites and pathway markers using chiral chromatography and tandem mass spectrometry

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