Ammonium acetate nh4oac

Acetonitrile (ACN), methanol (MeOH), and ammonium acetate (NH₄Oac), all LC–MS grade, were purchased from Fisher Scientific (Pittsburgh, PA). Ammonium hydroxide (NH₄OH), O-methylhydroxylamine hydrochloride (MeOX), and N-Methyl-n-(tert-butyldimethylsilyl) trifluoroacetamide (MTBSTFA) were bought from Sigma-Aldrich (Saint Louis, MO). Deionized water was provided in-house by a water purification system from EMD Millipore (Billerica, MA). Phosphate buffered saline (PBS) was bought from GE Healthcare Life Sciences (Logan, UT). Standard compounds corresponding to measured aqueous metabolites/features were purchased from Sigma-Aldrich and Fisher Scientific. Lipid standards were purchased from Fisher Scientific, Sigma-Aldrich, and Avanti Polar Lipids (Alabaster, AL).

Potassium dihydrogen phosphate (KH₂PO₄), sodium dihydrogen phosphate (NaH₂PO₄), L-aspartic acid (catalog #A93100, batch #12002LC), L-glutamic acid (catalog #G1251, lot #SLBK8671V), magnesium chloride, and oligonucleotides were purchased from Sigma-Aldrich (St. Louis, MO, USA). The radiolabeled reagents aspartic acid, L-[2,3-3H] (catalog #ART 0211, lot #180606), glutamic acid, L-[2,3,4-3H] (catalog #ART 0132, lot #180710) and adenosine triphosphate (ATP) [γ-³²P] were purchased from American Radiolabeled Chemicals (St. Louis, MO, USA). Ampicillin, chloramphenicol, kanamycin, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), ethylenediaminetetraacetic acid (EDTA), isopropyl β-d-thiogalactopyranoside (IPTG), phenylmethanesulfonyl fluoride (PMSF), and tris(hydroxymethyl) aminomethane (Tris) were from GoldBio Biotechnology, Inc., (St. Louis, MO, USA). Potassium chloride (KCl), sodium chloride (NaCl), trichloroacetic acid (TCA), and ammonium acetate (NH₄OAc) were purchased from Fisher Scientific (Hampton, NH, USA).

Optima grade methanol (MeOH), acetonitrile (ACN), isopropanol (ISP), and acetic acid (AcOH, > 98% pure) were purchased from Fisher Scientific (Pittsburg, PA, USA). UHPLC-grade methyl tert-butyl ether (MTBE) and hexane were also purchased from Fisher Scientific (Pittsburg, PA, USA). Methyl formate (> 98% pure) was purchased from Acros Organics (Morris Plains, NJ, USA). Ammonium acetate (NH₄OAc) was purchased from Fisher Scientific (Fair Lawn, NJ, USA). Deionized water was obtained through filtration via a Millipore Q-Plus (Elix^®35, Molsheim, France). For study 1, native SPM standards: resolvin D1 (RvD1), resolving D2 (RvD2), resolving D3 (RvD3), resolvin D5 (RvD5), resolving E1 (RvE1), maresin (MaR1), and protectin (PD1), and deuterated SPM standards: resolvins (RvD1-d₅, RvD2-d₅, RvD3-d₅, RvE1-d₄) maresin (MaR1-d₅), and leukotriene B4-d₄ were purchased from Cayman Chemicals (Ann Arbor, MI, USA). C18 SPE cartridges (Supel^TM-Select HLB SPE 30 mg loading, 1 mL volume) were purchased from Supelco Analytical (Bellefonte, PA, USA). For study 2, lipid standards were purchased from Cayman Chemical (Ann Arbor, MI, USA), Santa cruz Biotechnology, Inc (Dallas, TX, USA) and Avanti polar Lipids, Inc (Alabaster, AL, USA), as summarized in Supplementary Materials: Table S1.