Formic acid, tert-butyl methyl ether (MTBE), ammonium formate, ammonium acetate, acetate acid (HAc), sodium chloride, sodium bicarbonate, potassium chloride, glucose, disodium phosphate, HEPES, and calcium chloride were purchased from Sigma Aldrich (Steinheim, Germany). The ULC/MS-grade solvents, acetonitrile (ACN), methanol (MeOH) were obtained from Biosolve (Valkenswaard, Netherlands) and isopropanol (IPA) was purchased from Merck (Darmstadt, Germany). Ultrapure water (18 MΩ cm at 25 °C) was used to prepare solutions. Sodium dodecyl sulfate (SDS) was obtained from Roth (Karlsruhe, Germany), tris(hydroxymethyl)-aminomethane (Tris) from Applichem (Darmstadt, Germany), and sodium chloride (NaCl) from Merck (Darmstadt, Germany). Platelets were activated using collagen-related peptide (Richard Farndale, University of Cambridge, United Kingdom) or thrombin from human plasma (Roche, Germany).
One hundred and thirty-six lipid standards (60 mediators, 58 thereof selected after model review) (See Supplementary Methods) were used to study the lipid fragmentation, build optimal collision-energy models, create in silico spectral libraries and spike-in as internal standards. They were purchased from Avanti (Alabaster, AL, USA) and Cayman Chemical (Ann Arbor, Michigan, USA). Lipid fragment masses of mediators were validated with the Metlin database (https://metlin.scripps.edu/ )66 (link).
One hundred and thirty-six lipid standards (60 mediators, 58 thereof selected after model review) (See Supplementary Methods) were used to study the lipid fragmentation, build optimal collision-energy models, create in silico spectral libraries and spike-in as internal standards. They were purchased from Avanti (Alabaster, AL, USA) and Cayman Chemical (Ann Arbor, Michigan, USA). Lipid fragment masses of mediators were validated with the Metlin database (