5 s hete d8

HPLC-grade water, methanol, acetonitrile, and isopropyl alcohol were purchased from J.T. Baker (Avantor Performance Material, Inc., Center Valley, PA, USA). Acetic acid and ammonium acetate were obtained from Sigma-Aldrich (St. Louis, MO, USA). The eicosanoid standards were as follows: 14,15-EET-d11, 5(S)HETE-d8, LTB₄-d4, PGE₂-d4, PGD₂-d4, and AA-d8 (Cayman Chemical, Ann Arbor, MI, USA). A Strata-x 33-µm polymerized solid reverse-phase extraction column (cat # 8B-S100-UBJ) was purchased from Phenomenex (Torrance, CA, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin, and trypsin-ethylenediaminetetraAcetic acid (EDTA) were purchased from HyClone Laboratories Inc. (Logan, UT, USA). Escherichia coli LPS and Griess reagent were obtained from Sigma Chemical Co. (St Louis, MO, USA).

Eicosanoids; 5(S)-HETE, 12(S)-HETE, 15(S)-HETE, and AA were purchased from Cayman Chemical Co. (Ann Arbor, Michigan, USA). 5(S)-HETE-d8 (Cayman Chemical Co) was used as an internal standard (IS) for this study. Acetonitrile (Baker analyzed® LC-MS reagent), water (Baker analyzed® LC-MS reagent), formic acid, ethanol, ethyl acetate (EtOAc), and glacial acetic acid were used in sample preparations for LC–MS/MS measurements.

Lidocaine (2%) and proxymetacaine (0.5%) eye drops were from Moscow Endocrine Plant (Moscow, Russia) and Alcon-Couvreur (Puurs, Belgium), respectively. The Schirmer test tear strips were from Haag-Streit (Bern, Switzerland). Butylated hydroxytoluene (BHT) was from Sigma-Aldrich (St. Louis, MO, USA). The deuterated oxylipins standards 6-keto PGF1α-d4, TXB2-d4, PGF2α-d4, PGE2-d4, PGD2-d4, LTC4-d5, LTB4-d4, 5(S)-HETE-d8, 12(S)-HETE-d8, 15(S)-HETE-d8, Oleoyl Ethanolamide-d4, EPA-d5, DHA-d5 and AA-d8 were from Cayman Chemical (Ann Arbor, MI, USA). Solid-phase lipid extraction cartridge Oasis^® PRIME HLB was obtained from Waters (Eschborn, Germany). Other chemicals were from Sigma-Aldrich, Amresco (Solon, OH, USA), or Serva (Heidelberg, Germany) and were at least reagent grade.