title: Metabolomics profiling of the free and total oxidised lipids in urine by LC-MS/MS: application in patients with rheumatoid arthritis.
abstract: Oxidised lipids, covering enzymatic and auto-oxidation-synthesised mediators, are important signalling metabolites in inflammation while also providing a readout for oxidative stress, both of which are prominent physiological processes in a plethora of diseases. Excretion of these metabolites via urine is enhanced through the phase-II conjugation with glucuronic acid, resulting in increased hydrophilicity of these lipid mediators. Here, we developed a bovine liver-β-glucuronidase hydrolysing sample preparation method, using liquid chromatography coupled to tandem mass spectrometry to analyse the total urinary oxidised lipid profile including the prostaglandins, isoprostanes, dihydroxy-fatty acids, hydroxy-fatty acids and the nitro-fatty acids. Our method detected more than 70 oxidised lipids biosynthesised from two non-enzymatic and three enzymatic pathways in urine samples. The total oxidised lipid profiling method was developed and validated for human urine and was demonstrated for urine samples from patients with rheumatoid arthritis. Pro-inflammatory mediators PGF2α and PGF3α and oxidative stress markers iPF2α- IV, 11-HETE and 14-HDoHE were positively associated with improvement of disease activity score. Furthermore, the anti-inflammatory nitro-fatty acids were negatively associated with baseline disease activity. In conclusion, the developed methodology expands the current metabolic profiling of oxidised lipids in urine, and its application will enhance our understanding of the role these bioactive metabolites play in health and disease.
authors.affiliation: Department of Analytical Biosciences, Leiden Academic Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands. j.fu@lacdr.leidenuniv.nl.|Sino-Dutch Center for Preventive and Personalized Medicine, P.O. Box 360, 3700 AJ, Zeist, The Netherlands. j.fu@lacdr.leidenuniv.nl.
authors.forename: Junzeng
authors.initials: J
authors.lastname: Fu
authors.affiliation: Department of Analytical Biosciences, Leiden Academic Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.|Netherlands Metabolomics Centre, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.
authors.forename: Johannes C
authors.initials: JC
authors.lastname: Schoeman
authors.affiliation: Department of Analytical Biosciences, Leiden Academic Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.|Netherlands Metabolomics Centre, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.
authors.forename: Amy C
authors.initials: AC
authors.lastname: Harms
authors.affiliation: Sino-Dutch Center for Preventive and Personalized Medicine, P.O. Box 360, 3700 AJ, Zeist, The Netherlands.|TNO, Netherlands Organization for Applied Scientific Research, Microbiology and Systems Biology, P.O. Box 360, 3700 AJ, Zeist, The Netherlands.
authors.forename: Herman A
authors.initials: HA
authors.lastname: van Wietmarschen
authors.affiliation: Department of Analytical Biosciences, Leiden Academic Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.|Netherlands Metabolomics Centre, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.|Discovery Sciences, Janssen R&D, Turnhoutseweg 30, 2340, Beerse, Belgium.
authors.forename: Rob J
authors.initials: RJ
authors.lastname: Vreeken
authors.affiliation: Department of Analytical Biosciences, Leiden Academic Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.|Netherlands Metabolomics Centre, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.
authors.forename: Ruud
authors.initials: R
authors.lastname: Berger
authors.affiliation: Rheumatology and Clinical Immunology, University Medical Center Utrecht, F02.127, Heidelberglaan 100, 3584 CX, Utrecht, The Netherlands.
authors.forename: Bart V J
authors.initials: BV
authors.lastname: Cuppen
authors.affiliation: Rheumatology and Clinical Immunology, University Medical Center Utrecht, F02.127, Heidelberglaan 100, 3584 CX, Utrecht, The Netherlands.
authors.forename: Floris P J G
authors.initials: FP
authors.lastname: Lafeber
authors.affiliation: Department of Analytical Biosciences, Leiden Academic Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.|Sino-Dutch Center for Preventive and Personalized Medicine, P.O. Box 360, 3700 AJ, Zeist, The Netherlands.|Netherlands Metabolomics Centre, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.|TNO, Netherlands Organization for Applied Scientific Research, Microbiology and Systems Biology, P.O. Box 360, 3700 AJ, Zeist, The Netherlands.
authors.forename: Jan
authors.initials: J
authors.lastname: van der Greef
authors.affiliation: Department of Analytical Biosciences, Leiden Academic Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.|Netherlands Metabolomics Centre, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hankemeier
mesh_terms: D000328:Adult; D000818:Animals; D001172:Arthritis, Rheumatoid / Q000378:metabolism* / Q000652:urine*; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods; D004926:Escherichia coli / Q000201:enzymology; D005260:Female; D005966:Glucuronidase / Q000378:metabolism; D006372:Helix, Snails / Q000201:enzymology; D006801:Humans; D006868:Hydrolysis; D050356:Lipid Metabolism; D008055:Lipids / Q000652:urine*; D008297:Male; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D008055:Lipids; D005966:Glucuronidase
keywords: LC-MS/MS; Metabolomics; Oxidized lipids; Urine; β-glucuronidase