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2 protocols using sphingolipids

1

Sourcing Analytical Standards for Metabolomics

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Methanol and chloroform were purchased from Biosolve Chimime SARL (Dieuze, France). The 1-butanol was purchased from Acros Organics (Geel, Belgium). Butylated hydroxytoluene (BHT) and methyl tert-butyl ether (MTBE), and citric acid and sodium dihydrogen phosphate dehydrate were obtained from Sigma-Aldrich (Steinheim, Germany). MilliQ water was obtained from a Millipore high-purity water dispenser (Billerica, MA, USA). All solvents were HPLC grade or higher.
For internal standards (ISTDs), deuterium-, carbon-, and/or nitrogen-labelled metabolites were used. Labelled oxylipins, fatty acids, and endocannabinoids ISTDs were acquired from Cayman Chemicals (Ann Arbor, MI, USA). Labelled lysophospholipids, sphingolipids, and bile acid and steroid ISTDs were ordered from Avanti Polar Lipids (Alabaster, AL, USA). Labelled amino acids and amine ISTDs were ordered from Cambridge Isotope Laboratories (Andover, MA, USA), and labelled ATP, AMP, and UTP were purchased from Sigma-Aldrich (Steinheim, Germany).
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2

Lipid Mediator Profiling Protocol

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Eicosanoids and endocannabinoids were purchased from Cayman Chemical (Ann Arbor, USA), with the exception of 12(13)-EpODE, 9,10,13-TriHOME and 9,12,13-TriHOME from Larodan (Solna, Sweden) and LTB5 from Enzo (Solna, Sweden). Sphingolipids were purchased from Avanti Polar Lipids (Alabaster, USA). A complete list of lipid mediator nomenclature is provided in Additional file 2. Methanol, acetonitrile, isopropanol, acetic and formic acid were of LC-MS grade from Fischer Scientific (Stockholm, Sweden). Ethyl acetate and chloroform were purchased from Sigma-Aldrich (St. Louis, USA).
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